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https://openalex.org/W4320024969	https://scholarworks.iu.edu/journals/index.php/josotl/article/download/32702/38725	English	null	Bringing Face-to-Face Engagement to Online Classes	The journal of scholarship of teaching and learning	2,022	cc-by	9,773	Bringing Face-to-Face Engagement to Online Classes: Developing a High-Presence Online Teaching Method Gregory Gimpel J. Mack Robinson College of Business Georgia State University ggimpel@gsu.edu Abstract: The trend to shift courses online is accelerating. Some students are gravitating toward asynchronous online classes; however, many still prefer in-person educational experiences. These students often are less engaged when taking online courses, and their willingness to pay for online courses is frequently less than for in-person courses. There is a need to bridge the gaps between online and traditional class delivery formats. This paper reports on a high-presence online teaching method that approximates the in-person experience by affording face-to-face conversations, real-time interaction, and features the instructor placed front-and-center with the lecture material. The paper reports a case study test of this method as applied to a graduate process reengineering course. Students in the course report that the method provides better student-instructor interaction and overall engagement than they expect from in-person classes. Students also report that the method approximates what they expect from in-person courses regarding the quality of interactions they have with their classmates. Keywords: Online education, student engagement, social presence, media richness, instructional technology, course design, experiential learning, synchronous online distance learning. While the number of online classes has been increasing, particularly over the past decade, externa forces, especially COVID-19, have accelerated the transition to online teaching formats. Man students have struggled staying engaged with their courses during the shift to online, remote classes. While the number of online classes has been increasing, particularly over the past decade, external forces, especially COVID-19, have accelerated the transition to online teaching formats. Many students have struggled staying engaged with their courses during the shift to online, remote classes. Although some students favor online learning, research suggests that most students prefer face-to-face instruction (Jaggars, 2014) and that many students view online courses as less desirable compared to face-to-face courses (Tichavsky, Hunt, Driscoll, & Jicha, 2015). Of students surveyed, 50% found that online courses lacked sufficient interaction with their instructor, 26% indicated that online courses lacked immediate instructor feedback as a critical missing component, and 10% indicated difficulty in getting clarification of material because they could not interact with the instructor. Other students noted the absence of the instructor’s energy and enthusiasm during online instruction. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022, pp.32-49. doi: 10.14434/josotl.v22i4.32702 Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022, pp.32-49. doi: 10.14434/josotl.v22i4.32702 Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022, pp.32-49. doi: 10.14434/josotl.v22i4.32702 Bringing Face-to-Face Engagement to Online Classes: Developing a High-Presence Online Teaching Method Student satisfaction with their professors tend to be lower with online courses (Baker, 2010; Cole, 2016), frequently caused by a reduced amount of interpersonal interaction and personalized feedback from the instructor (Cole, 2016; Jaggars, 2014). Many students consider the quality of conventional online interactions such as discussion forums as lower than the quality of interactions that occur within a classroom (Tichavsky et al., 2015). This translates into students earning lower grades in online courses than in the traditional classroom setting (Bettinger, Fox, Loeb, & Taylor, 2017). For many students, their willingness to pay is much lower for online education, in part because some universities offer online degrees for a fraction of the price of comparable in-person degrees, but also because these students believe their educational experience is diminished with the reduction of face-to-face educational opportunities. A recent survey finds that 75% of students do not believe online classes provide a quality learning experience. Another survey finds that 67% of students find Gimpel Gimpel online classes less effective than in-person courses (Daniels, 2020). Student protests against the loss of in-person class options have grown louder, expressing resistance with statements like “There’s no need to pay out-of-state tuition if I’m at home” and “I’m not paying full price for YouTube university” (Icon By The Noun Project, 2020). Students are demanding tuition cost reductions when required to take online classes rather than in-person ones (O'Brien, 2020) and institutions of higher education are facing student lawsuits demanding tuition refunds and changes in tuition rates during the pandemic because online classes are fundamentally a different experience than in-person, face-to-face instruction (G. Anderson, 2020; Binkley, 2020). As health concerns abate and universities return to full capacity in their classrooms, funding, real estate costs, and other non-pedagogical factors will still drive a shift to online courses. At the same time, students will still maintain their expectations for interpersonal interaction, real-time engagement, and instructor motivation. This raises a key research question for teaching: how can the face-to-face benefits of in-person classes be captured while teaching an online class? In an attempt to answer this question, a high-presence online teaching method was developed for and tested during Summer 2020 for a process reengineering core taught within a masters of information systems program. Bringing Face-to-Face Engagement to Online Classes: Developing a High-Presence Online Teaching Method The method uses the Zoom video conferencing app to enable real-time, (digitally) face-to-face interaction among instructor and all students, while superimposing instructors onto lecture slides so that instructors can interact with the students and the content while delivering the lecture, capturing the rich interactions students expect when everyone is physically in the same class room. The results of this case study (Yin, 2017) indicate that much of the interaction and engagement of in-person classes can be replicated online. The online method tested in this study achieved greater student-faculty engagement and interaction than students expect from face-to-face classes and student-student interaction which closely approximated the expectations from face-to-face courses. Review of Related Literature Students’ desire for interaction and need to be motivated by the professor are key factors for creating effective learning environments (Tichavsky et al., 2015). These factors are often described as engagement, which can be increased through the use of “rich media” that approximates in person, face-to-face communication. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu Social Presence Social presence theory (Short, Williams, & Christie, 1976) suggests that immediacy and intimacy, conveyed by verbal cues and nonverbal cues such as facial expression and body language define one’s “social presence.” Social presence is the degree to which students feel emotionally and socially connected to others in the class (He, Xu, & Kruck, 2019). Social presence is about being actively engaged, but more importantly, perceived as being “real,” understanding what makes a person authentic and unique, and “there,” in a supporting and caring sense (Lowenthal & Snelson, 2017). Social presence is a necessary component of engagement (Dixson, 2010). Higher immediacy and intimacy create strong social presence and foster stronger connections (Keil & Johnson, 2002). In a computer-mediated setting, it is important for teams to project themselves socially into a community of inquiry (Lowenthal & Snelson, 2017). Specific to the teaching setting, instructor presence is the visibility of the teacher as perceived by the student (Baker, 2010). Instructor presence includes facilitating necessary discussions and maintaining student engagement. It builds the connectedness between teacher and students required for strong engagement (T. Anderson, Liam, Garrison, & Archer, 2001; Baker, 2010). Face-to-face instruction has higher social presence than computer- mediated channels; however, media that offer increased immediacy, intimacy, and instructor visibility can offer a high degree or social presence (Keil & Johnson, 2002). Computer mediated communication offers a spectrum ranging from lean to rich (Lowenthal, 2010). As such, the media richness of the technology tools directly affects instructor social presence and student engagement. Engagement Student engagement is the most critical factor for student learning (A. Singh, Rocke, Pooransingh, & Ramlal, 2019) and satisfaction with an online course (Martin & Bolliger, 2018). Student engagement can be summarized as students’ psychological investment and effort focused toward learning and mastering the knowledge and skills taught in a class (Lamborn, Newmann, & Wehlage, 1992). Students are more engaged when the instruction increases contact between students and teacher, provides timely feedback on student’s work, provides an opportunity for students to work collaboratively, and establishes high standards for what constitutes acceptable quality work. These course characteristics can be diminished or lost when courses move online, making students’ psychological investment and effort tenuous when courses are moved online (Martin & Bolliger, 2018). There are three types of interactions necessary for effective engagement and interaction in online courses: learner-to- instructor, learner-to-learner, and learner-to-content (Moore, 1989). Learner-to-instructor engagement is the most important for successful online education. The two most important factors for fostering learner-to-instructor engagement is timely instructive feedback and the instructor’s interaction (i.e. presence). Learner-to-content engagement can be achieved though “authentic activities” such as real- world activities. Group work builds learner-to-learner engagement (Martin & Bolliger, 2018). 33 Gimpel Immediacy Interaction is a cornerstone of the learning experience and strongly shapes learning outcomes (Wanstreet, 2009). Interaction impacts student motivation, active learning, and successful achievement of learning objectives. Opportunities for students to interact with each other and the instructor are necessary for effective online instruction (Dixson, 2010). Interactive immediacy, such as asking and answering questions, calling students by name, and communicating attentiveness to students, is necessary for effective learning (Baker, 2010). The ability for students to interrupt the instructor, such as to ask a question or request clarification, is important to learning new material and student engagement (Y. C. Liu & Burn, 2007). Immediacy and real-time interaction, such as those afforded by synchronous activities in an online course, positively impact student affective learning, motivation, and cognition (Baker, 2010), whereas asynchronous online instruction can result in student disengagement (Romero-Hall & Vicentini, 2017). Immediacy is an antecedent of social presence. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu 34 Media Richness Online media afford various ways to increase visibility and instructor presence, creating different opportunities to provide verbal and visual cues that will engage students. Daft and Lengel (Daft & Lengel, 1984) propose a continuum between rich and lean media, based on a medium’s ability to convey a broad or a limited range of cues. The theory posits that a high task-medium fit in communication media provides the ideal range of cues. Media richness has a positive impact on student concentration and satisfaction with online learning (S.-H. Liu, Liao, & Pratt, 2009). Commonly used “lean” media used in online education, such as chat boxes and widgets to replicate raising hands provide a very limited range of cues. On the other hand, media with high media richness afford teachers and students to communicate more effectively, including picking up on non-verbal cues (Fernandez, Simo, Sallan, & Enache, 2013; Ferschke, Yang, Tomar, & Rosé, 2015; S.-H. Liu et al., 2009; Romero-Hall & Vicentini, 2017). High media richness can reduce the distance among learning 34 Gimpel Gimpel participants, even when they are geographically separated (Zhao, Wang, & Sun, 2020). A medium is considered to be richer when it more closely it replicates face-to-face communication. Rich media are best suited for teaching and learning new material (Cole, 2016). participants, even when they are geographically separated (Zhao, Wang, & Sun, 2020). A medium is considered to be richer when it more closely it replicates face-to-face communication. Rich media are best suited for teaching and learning new material (Cole, 2016). Online Teaching Method The goal driving the redesign of the course was to capture many of the face-to-face benefits of in- person classes in an online class by matching the in-person, face-to-face experience as closely as possible. To accomplish this goal, a Summer 2020 process reengineering course employed the richest media available given budgetary constraints and the limitations of setting up a teaching studio in the basement of a suburban home. The media capabilities are modeled on the “WOW Rooms” at schools such as IE and Georgia State University (IE University, 2020; J. Mack Robinson College of Business, 2020), but with a budget of under $1000. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl indiana edu Lecture Design The course design employed 4.25 hour long, live lectures on Saturday mornings. (This is the same length and schedule that in-person classes are usually offered. The class duration and schedule was a holdover from the schedule created by the registrar before the course moved online.) Attendance was required for all students. The class sought to replicate the instructor presence of an in-person course. Like in-person classes in this program, copies of lecture slides were available to students before class and the professor speaks extemporaneously rather than from a prepared script. In most in-person classes, the instructor stands and uses a PowerPoint slideshow to accompany the lecture. The professor does more than simply provide a narration of a slide show. The instructor also monitors the facial reactions and body language of students, using real-time feedback to gauge engagement and whether students are grasping the material. Students answer questions posed by the instructor and can interrupt the lecture with questions and comments. y p q To replicate this class experience, the instructor stood in front of a green screen, which enabled the professor’s body to be superimposed over the lecture slides. Hence, the lecture was delivered similar to how a weatherman presents the weather on television. Most lecture slides were reformatted so that the instructor could stand beside the content rather than blocking it. This enables the instructor to point to images and bullet points on the slide during the lecture, replicating the in-class experience. Instead of writing on a white board with markers, the instructor uses a virtual “whiteboard” and a drawing tablet to write and draw content for the students. Creating lecture content in real-time provides a sense of immediacy. Students can engage directly with the lecture content by contributing the data that is written and drawn by the professor. This live conceptual elaboration is only possible because of the instructor’s presence, unlike information presented from a previously prepared lecture slide. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu 35 35 Gimpel Figure 1. Image of online class lecture. Figure 1. Image of online class lecture. To monitor student engagement, and so the professor could benefit from the non-verbal feedback from students, all students are required to log in with their webcams on. Lecture Design All of the 41 students webcam streams appear simultaneously on a 50-inch-high-definition monitor, so that the professor could see the entire class at the same time. Students are free to unmute themselves at any time to ask questions, make comments, etc. The chat feature of the online tool is disabled because 1) such a feature would not be used in an in-person class and 2) this communication channel would distract attention away from the richer audio-video medium, likely reducing engagement. Table 1. Summary of Online Lecture Design. Component Description Pedagogical Benefit Synchronous Teaching All students are required to attend each lecture, which is conducted live during a regularly scheduled time Allows for the real-time interaction and immediate answers to questions, both of which strongly shape learning outcomes Webcams Required All students are required to keep their webcams during the class. Images of all students are displayed to instructor concurrently on large monitor Increases instructor presence by enabling instructor to respond to facial expressions and body language of students Increases social presence of each student, because they can see each other in a window of the Zoom app Audio Communication Students make comments and ask questions using computer audio, not typing text chat Other communication channels would distract attention away from richer audio- visual medium, likely reducing engagement Text chat lacks the interruptibility that is important for learning new material and for student engagement “Weatherman” Effect Professor image is superimposed over lecture slides Increases instructor presence because the instructor is featured alongside the content 36 36 Gimpel Enables instructor to point to images during instruction, increasing interactivity and increasing engagement Virtual White Board Instructor writes and draws images on virtual white board Increases immediacy because content creation can be interactive, using student- supplied ideas and data, in real-time discussions Using student-supplied information increases interactivity and students’ engagement with the content and the instructor It is not pre-created PowerPoint animation, but conceptual elaboration that exists only because of the instructor’s presence Another large monitor allows the instructor to see what is broadcast to the students – the instructor’s image superimposed on top of a slide. This keeps instructors on point and also enables instructors to see themselves when gesturing to content on a slide and to make sure that the content is visible to the students. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl indiana edu Lecture Design This helps provide a quality control check both for content visibility and for the instructor’s interaction with the on-screen content. Prior to the first class, students received an orientation video link (Dunlap & Lowenthal, 2014). The video explained the format of the class. It also instructed students how to create a Zoom account using their university assigned email address. Email-based Zoom accounts were needed so that students could be sent to pre-assigned breakout rooms for in-class activities in which students would work together in teams. Figure 2. Screen shots from video introduction. Video instructions to class explaining the use of green screen and drawing tablet (left) and that one monitor would display the image of professor superimposed over lecture slides and the other monitor would show all students’ live webcam videos (right). Figure 2. Screen shots from video introduction. Video instructions to class explaining the use of green screen and drawing tablet (left) and that one monitor would display the image of professor superimposed over lecture slides and the other monitor would show all students’ live webcam videos (right). Video instructions to class explaining the use of green screen and drawing tablet (left) and that one monitor would display the image of professor superimposed over lecture slides and the other monitor would show all students’ live webcam videos (right). Video instructions to class explaining the use of green screen and drawing tablet (left) and that one monitor would display the image of professor superimposed over lecture slides and the other monitor would show all students’ live webcam videos (right). Assignments Assignments Historically this course has been a high-touch class, with much instructor-student interaction and significant interaction among the students. It employs an active learning philosophy in which students Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu 37 37 Gimpel Gimpel interact closely with the material, other students, and with the teacher (Bellanca, 2009; Demirci & Düzenli, 2017). Student-to-student interaction is fostered by team homework assignments, ungraded in-class team activities, and a term-long team project. The project follows an active learning technique employing a term-long group project with multiple milestones and includes in-class student group activities during most class sessions (Dadashzadeh, 2018; Gudigantala, 2013). This type of cooperative learning helps advance student understanding of the material (Ott, Carpenter, Hamilton, & LaCourse, 2018). For the past few years, students have been given an assignment that is due before the first class. The assignment requires students to watch instructional videos and submit a BPMN process model to get credit. The first assignment engages students with the course material prior to the first class. Students are required to complete a business process modeling assignment on their own. Each student receives detailed, customized feedback. The personalized feedback can maintain instructor presence and a connection with each student throughout the semester (Dunlap & Lowenthal, 2014). The feedback also requires students to revisit the assignment, increasing student-content interaction. q g g The course is built around an in-depth process reengineering project based on processes that the students use at their jobs. Projects are selected based on the scope and suitability of a process for accomplishing the learning objectives within the 8-week “minimester” term. The first and second class sessions, along with several other sessions, involve breaking students into their teams to complete in- class activities based on the topics of the day. These structured small group collaborations enhance student-to-student engagement (Dunlap & Lowenthal, 2014; Lowenthal, 2009). The nature of the online method requires that breakout rooms be assigned prior to the beginning of a class session. As such, student teams need to be identified prior to the first day of class. Therefore, students were required to post to a discussion forum information about the business processes. The instructor selected the most suitable processes, then assigned students to teams based on those processes prior to the first class. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl indiana edu Assignments Like the first homework assignment, the discussion post requires thought and engages the student with the course topic before the first class. p The course contains in-class team activities, in which the students apply the lecture content to an ungraded assignment. Students are sent into virtual breakout rooms to perform these assignments. The instructor visits each breakout room to check on the students and answer any questions they may have. The students then return to the main room and discuss the work they did. Teams are required to submit their work, even though they do not receive a grade. g g The team project is an in-depth examination and redesign of a process at a real-world company, following active learning principles established by Merrill (2002). The problem solved by students is authentic. The project consists of multiple interim deliverables, applying a scaffolding approach (Demetriou, Spanoudis, & Mouyi, 2011; Paas, 1992) so that students can apply new knowledge to existing knowledge throughout the course. The instructor meets with each team to provide feedback after each deliverable, allowing the demonstration of knowledge and the contextual integration of that knowledge into the project. Student teams schedule a meeting time from a menu of available times. Even for the previous face-to-face versions of this course, these meetings were conducted over video conference. This remained the same when the class was converted to a 100% online course. This type of project-based learning has been shown to increase student participation, increase understanding of course material, master the skills taught in a class, and increase enthusiasm for self-directed learning (Dadashzadeh, 2018). 38 Gimpel Table 2. Summary of Assignments. Component Description Pedagogical Benefit Homework Assignment Due Before First Lecture Students learn foundational concepts via prerecorded videos and apply them to an assignment before first class Builds learner-to-content engagement prior to first lecture Individual Assignment Students create a BPMN 2.0 process diagram based on a written description of the process. Builds learner-to-content engagement. Individualized feedback for each student increases instructor presence. In-Class Assignments (via pre-assigned groups & breakout rooms) Students are assigned to breakout rooms to work on ungraded assignments Enables students to apply lecture content to practical problems, increasing learner-to-content engagement. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl indiana edu Assignments Increases learner-to- learner engagement by allowing students to work collaboratively Helps students identify gaps in understanding which can be discussed with rest of class and with instructor Group Homework Students complete 2 homework assignments as a team Builds learner-to-learner and learner- to-content engagement Team Project Students apply comprehensive knowledge of course topics to solve real-world problem Professor meets with teams multiple times to provide feedback Builds learner-to-content and learner- to-learner engagement Increases instructor presence, fostering learner-to-instructor engagement ble 2. Summary of Assignments. Pedagogical Benefit Figure 3 includes screen shots of the prerecorded introductory video with an explanation of the discussion board assignment (left) and the first technical assignment (right), both of which were due before the first class. Figure 3. Screen shots of explanation of initial assignments. Figure 3. Screen shots of explanation of initial assignments. 39 39 Gimpel Gimpel Instructional Technology The use of technology was inspired by the state-of-the-art classrooms that allow an instructor to interact with all the students in the class in real-time, despite being in different locations. Instead of a wall of large monitors, one 50” high-definition television was used. It was placed about 2 meters away from the instructor. The video conferencing software used was Zoom, which allows for 49 people to be displayed on a screen at one time. If more than 49 people attend the class, the instructor can scroll through different screens, each of which will display 49 other people. Despite 41 student images (the number of students enrolled) appearing on the screen at the same time with their names accompanying the images, the student images were clear and it is easy to read their facial expressions and body language. The laptop speakers were used to hear students and the instructor spoke into a USB- connected wireless lapel microphone. JPG images of the lecture slides are uploaded as Zoom background images. During the lecture, they are controlled on the laptop, using the cursor keys to advance background slides. It takes a few minutes to upload the slides. Each one must be uploaded individually, but it only takes 3 clicks of the mouse for each one. Forty or fifty slides can be added in about 10 minutes. At the time of this case study, a solid colored background was required for the “weatherman effect.” Flat lighting provides the best effect. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu Assignments The basement setting used in this pilot has uneven lighting, so photographer’s umbrella lights are used to provide appropriate lighting for green screen effects. A cloth green screen hung behind the instructor, although it would not be needed if the room had a single-colored wall and flat lighting. (Advances in Zoom’s technology no longer require a green screen or good lighting for the weatherman effect.) A high definition webcam is on a tripod in front of the instructor. The monitor displaying the broadcast image is immediately behind the webcam, so instructors looks directly at the webcam at the same time they see the image broadcast to students. Figure 4. Studio set-up (Located in residential basement). Figure 4. Studio set-up (Located in residential basement). Zoom sessions were managed using a Dell Latitude with 16GB RAM and a Dell DisplayLink, which allows connecting the laptop to 2 external monitors. The image of the instructor and slides appeared on one external monitor. The faces of all the students were on the other external monitor. 40 40 Gimpel Gimpel The Zoom audio and video controls, which include which background image is displayed, appeared on the laptop screen. The laptop and DisplayLink were issued by the College for general job duties. Excluding the laptop and DisplayLink, the total budget for the online studio was under $1000. The Zoom audio and video controls, which include which background image is displayed, appeared on the laptop screen. The laptop and DisplayLink were issued by the College for general job duties. Excluding the laptop and DisplayLink, the total budget for the online studio was under $1000. Table 3. Teaching Studio Components and Actual Price Paid. Teaching Studio Component Price Paid Sceptre 50" 4K UHD LED TV 199.99 Sceptre 50" 4K UHD LED TV 199.99 Kanto MTM65PL Mobile TV Stand 129.99 Kanto MTM65PL Mobile TV Stand 129.99 Linco Photo Video Studio Light Kit AM169 (Includes Green Screen) 119.99 Wacom Drawing Tablet 79.99 Microsoft 1080p LifeCam 52.95 Fifine K031 Wireless Lavalier Microphone with USB Receiver 39.99 Amazon Basics 6-Outlet, 6' Surge Protector Power Strip 10.99 Coleman Cale 7' Indoor Extension Cord 4.95 Amazon Basics 4K HDMI Cable 14.99 Amazon Basics 4K HDMI Cable 14.99 Total Cost 998.80 Table 3. Teaching Studio Components and Actual Price Paid. f Teaching and Learning, Vol. 22, No. 4, December 2022. y g g g y g Professor presents the material in a way that is easy to follow and understand the material Professor presents the material in a way that is easy to follow and understand the mat The biggest surprise is that student engagement was perceived as superior to in-person instruction. Given the substantial number of studies that find student engagement is less in online classes, the goal was to approximate a 3.0 score (i.e. “same” rating). Table 5 lists student feedback about students’ engagement with the course, its material, and the instructor. Table 6. Feedback about Student Engagement. Table 6. Feedback about Student Engagement. Professor is highly professional. He has a the ppt in background and is very successful in engaging the students. One of the best experiences and best class attended. I liked the fact that even though this class was 100% online the professor asked questions to students and made sure everyone was attentive You tried to imitate a classroom setting with the virtual background and seeing us as well, it made me personally more engaged to see that you wanted to deliver the best you could for us. The slides as background during the lectures and the professor standing in front. It helped with staying focused rather than just looking at a slide and hearing a professor talk without seeing them. Updates every week with the professor kept me engage within the class. g g Professor is highly professional. He has a the ppt in background and is very successful in engaging the students. One of the best experiences and best class attended. I liked the fact that even though this class was 100% online the professor asked questions to students and made sure everyone was attentive You tried to imitate a classroom setting with the virtual background and seeing us as well, it made me personally more engaged to see that you wanted to deliver the best you could for us. The slides as background during the lectures and the professor standing in front. It helped with staying focused rather than just looking at a slide and hearing a professor talk without seeing them. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu Table 5. Student Feedback about Student-Instructor Interactions. I really liked the way Professor arranged this online class in a very effective way, that no student misses anything and able to gauge everything. Data Collections & Results This class was almost similar to in-class experience. I really appreciate the efforts of the professor to make this class as interactive as possible. Even though the class was online, there was full engagement by the students and professor. The class required webcam to be on, so it was not dull. There was a lot of interaction. The webcam and green screen setup by the professor was really helpful and I appreciate this a lot which made this class much more interesting. I really liked the way Professor arranged this online class in a very effective way, that no student misses anything and able to gauge everything. Professor presents the material in a way that is easy to follow and understand the material Table 5. Student Feedback about Student-Instructor Interactions. The interactivity and the flow of the class. Along with that the practical examples of the topic in hand combined with the in-class exercises. This class was almost similar to in-class experience. I really appreciate the efforts of the professor to make this class as interactive as possible. Even though the class was online, there was full engagement by the students and professor. The class required webcam to be on, so it was not dull. There was a lot of interaction. The webcam and green screen setup by the professor was really helpful and I appreciate this a lot which made this class much more interesting. I ll lik d h P f d hi li l i ff i h d Data Collections & Results The class was taught during Summer 2020. Prior to this class, the students had completed 30 hours of master-level instruction, most of which were taught face-to-face in a lecture hall. This previous experience informed students’ expectations for face-to-face classes. At the end of the term, students were asked to complete a survey with 5 Likert-type questions and two open answer questions based on the teaching method used in the course. Participation was completely voluntary. One student did not participate. The 5 Likert-type questions were based on a 5-point scale: much less, less, same, more, much more. The instructions were as follows: The Likert-type questions were converted to numeric scores with 1 representing “much less,” 3 representing “same,” and 5 representing “much more.” The objective was to achieve a 3 (“same”) rating. Table 4. Student Evaluations of Class Format. Item Mean Stnd Dev Compared to my expectations of a face-to-face class, the ability to interact with the professor in this online class was 3.3 0.74 Compared to my expectations of a face-to-face class, ability to interact with classmates during class time was 2.7 0.76 Compared to my expectations of a face-to-face class, this class engaged my attention 3.6 0.90 Compared to my expectations of a face-to-face class, quality of interaction between students and professor in this online class was 3.2 0.95 Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. j tl i di d 41 Gimpel Compared to my expectations of a face-to-face class, quality of interaction between myself and classmates was 2.9 0.89 Compared to my expectations of a face-to-face class, quality of interaction between myself and classmates was 2.9 0.89 0.89 This lecture style increases student-instructor interaction and student engagement. The survey results indicate that students found both the ability to interact with the instructor and the quality of that interaction to be superior to those they expect to have in face-to-face course. This result is surprising, especially given that the students took this course at the end of their master’s program, after taking in-person classes from top professors. Table 4 lists qualitative feedback students provided about these interactions based on responses to what the students liked most about the class. Table 5. Student Feedback about Student-Instructor Interactions. The interactivity and the flow of the class. Along with that the practical examples of the topic in hand combined with the in-class exercises. Table 7. Feedback about Student-Student Interactions. Table 7. Feedback about Student-Student Interactions. It was the best online class I took here in [this university]. Especially the way we got to interact with our teams in the breakout rooms was something I enjoyed the most. I would like the class to have class interaction and collaboration between the students. The difficulty in working with groups online with Bizagi [the BPMN diagramming software used in the class] as one person has the program open and others have to point or state where to move a task or delete etc. When the process is complicated it can be difficult for everyone to know what area or task someone is talking about. Have more dialogue between the students to create more in depth conversation about other experiences. More Class discussions with professor and other students, interactive group discussions. Table 7. Feedback about Student-Student Interactions. It was the best online class I took here in [this university]. Especially the way we got to interact with our teams in the breakout rooms was something I enjoyed the most. I would like the class to have class interaction and collaboration between the students. The difficulty in working with groups online with Bizagi [the BPMN diagramming software used in the class] as one person has the program open and others have to point or state where to move a task or delete etc. When the process is complicated it can be difficult for everyone to know what area or task someone is talking about. Have more dialogue between the students to create more in depth conversation about other experiences. More Class discussions with professor and other students, interactive group discussions. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu Updates every week with the professor kept me engage within the class. The ability for students to interact with each other and the quality of those interactions approximated, but did not match the expectations students had for face-to-face classes. The ability interact scored a 2.7 and the quality of student-student interactions scored a 2.9, respectively representing 90% and 96.7% of what they expect from in-person classes. Four students wrote that “the thing I liked most” was the group work in the breakout rooms and another indicated that the best part of the method was “webcams and class participation.” Table 6 indicates longer statements 42 Gimpel provided by students regarding student-student interaction. Most of these comments provide constructive criticism. Discussions and Lessons The course delivery method piloted in this process reengineering course exceeded to the goal of matching the engagement and instructor-student interactions that students expect from in-person classes. While surprising, the data suggest that low-cost online teaching tools can translate into successful student experiences with online courses. This is very important given the likely increase in online teaching in the next few years. The overall goal of this pilot was to explore the research question: how can the face-to-face benefits of in-person classes be captured while teaching an online class? The results are encouraging, but this course approximated, but did not equal or exceed the expectations students have for student-student interactions. Technical Lessons Students were required to log in to Zoom using an account created using their university email address. There were two reasons for this policy. First, Zoom allows meeting hosts to preassign breakout rooms by uploading the email address of those assigned to each room. It is easy and convenient; however, it requires that meeting participants log in using an account associated with the email address specified in the breakout room assignments. Second, it makes sure that people log in using their names, not the name of a child or spouse who might share the computer. Seeing people’s faces and names on a monitor is similar to seeing students in a classroom with name placards at their desks. It helps the instructor and students match faces to names and learn who everyone is. Despite the instructions, each week some students entered the Zoom meeting without first logging in. This requires class time to manually assign them to breakout rooms. For this course, PowerPoint slides were converted to jpg files. A small percentage of the slides spanned the whole image. Most slides repositioned material so that it was on one side of the image and covered less than two-thirds of the image from left to right. That way the instructor’s image could be superimposed over the content without blocking it from view. Most of the slides contain large images and bold text written in a large font. Some slides, such as those depicting value stream maps or BPMN process diagrams, contain easy-to-view diagrams but small text. While Zoom generally does a good job delivering high-quality video images, small text may not reach participants clearly. Such images may need alterations to make the text larger so that it will be legible to the students. Also, if the internet speed drops to slow speeds, the background images lose focus and can become blurry. Care should be taken to make sure jpg versions of slides and other content use large text and large, sometimes simplified images. Additional lessons can be learned from other research into the use of Zoom for teaching online courses. A key lesson is that technology is a medium for teaching. Too much focus on the video conferencing application will lead to poor results. Some teaching experiments using Zoom were unsuccessful because they focused on the instructional technology rather than on the user experience (Stafford, 2020). In-Class Collaboration Lessons Most of the classes featured group work in which teams would spend 30-60 minutes working on a challenging, ungraded assignment. During this time, the instructor visits the different breakout rooms to answer any questions and provide advice. The purpose of these in-class activities has always been for students to engage in active learning by applying lecture concepts to real-world problems during class, so that they can realize what they do not understand. They can work with each other to gain an understanding, or raise their questions with the professor. Given that active learning group work takes longer when online than during in-person classes (de Oliveira Dias, Lopes, & Teles, 2020; Venton & Pompano, 2021), at least half an hour is needed in order for the professor to visit all the rooms, even just for a few minutes each. Longer breakout sessions provide more opportunity for interaction with other students and with the professor. The in-class group work serves to build mastery of concepts and material. At the same time, it is an opportunity for students to interact with each other. This online teaching method approximates the experience of the classroom, but it does not equal or exceed it. Students rate the ability to interact with each other as 2.7, which is 90% of what they expect in an in-person classroom. They rate the quality of those interactions as 2.9, which is 96.7% of what they expect from in person interactions. This data suggests that in an online class, some of the in-class group work should be redesigned to bolster student-student relationships and the quality of the interactions they have with each other. The current activities were designed with a general active learning approach (i.e. “dynamic type of learning where students intensely interact with each other, with educational materials, and with 43 Gimpel teachers” (Demirci, 2017)). Student-student interactions may be bolstered by employing specific active learning tools, rather than the current, more general approach. g g pp While this online teaching method engages students in small group discussions, the student- student interactions may be improved by employing a think-pair-share (TPS) approach, in which students work individually before solving the problem as a group, then present the group’s work to the class (McTighe & Lyman Jr, 1988). Beginning group activities with individual contemplation could prepare each student for more active engagement in the group discussion, and potentially provide more discussion and debate. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu In-Class Collaboration Lessons This could increase the quality of the interactions and also improve the ability to interact for students who prefer to take time to think before they speak. While TPS is frequently applied to short activities, it can be effective for longer, more detailed activities (Demirci & Düzenli, 2017). Instead of verbally asking students volunteers to speak their thoughts about various topics, “minute papers” could be adapted from the traditional use as an end-of-session reflection and feedback tool (Wilson, 1986) to a source of class discussion and participation (Holtzman, 2007). This would provide students a moment for reflection. The professor could then share and discuss a few of the minute paper responses. This would enable the students to engage more with their classmates’ ideas, and enable them to respond to classmates’ ideas, rather than be spectators as one student publicly discusses an idea with the instructor. Technical Lessons Technical Lessons The high-presence online teaching method detailed in this case study takes a holistic approach that rethinks lecture design and carefully constructs assignments. Technology is a key 44 Gimpel component, but in the high presence online teaching method, pedagogy and class design drove the choice of technology, not vice versa. Some teachers moved classes online with an a priori belief that an in-person classroom experience cannot be reproduced when delivered over communication software (de Oliveira Dias et al., 2020). Conversely, the method used for this course was created with the hope that a theory-based approach to course design could approximate the in-person experience. p y pp g pp p p Some studies that employed Zoom find that synchronous online class delivery can face many technical problems, such as those that result from limited broadband internet access among students (de Oliveira Dias et al., 2020) and the reality that many students may not have computers and other necessary equipment (C. K. S. Singh et al., 2020). Additionally, remote students can refuse to turn on their cameras, limiting the instructor-student and student-student interaction (Stafford, 2020). In the extreme, students can simply not attend online classes, although this can be addressed by requiring attendance as part of an overall grade, at least at the university level (de Oliveira Dias et al., 2020). While attendance, webcam use, and internet connectivity were not significant problems in this case study, this course was at the master level taught at an urban university. It is possible that enforcing webcam use may be more difficult in a secondary education environment. Courses with a base of rural students or schools located outside of major metropolitan areas with good broadband infrastructure may face technical difficulties using this high-presence online teaching method because it will be difficult for all students to transmit good quality video and audio. While all students in the course presented in this case study had laptops as a condition of enrollment, many universities, secondary, and primary schools may have students lacking the equipment needed to implement this teaching method. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl.indiana.edu Implications and Limitations Implications and Limitations The high presence online teaching method discussed in this case study can be applied to many different classes. The method is straight forward, with the lecture design, an assignment design that engages students before the start of the course, and engages them throughout the term through group work and in-class activities. While the subject matter was a highly-technical STEM subject within a master’s program, it should easily adapt to discussion-based courses within arts and humanities. The method provides a pedagogical and technical foundation that can be adapted based on the subject matter and the audience. At the same time, the results of this case study should be considered within the context of its limitations. The sample size (n=40) represented a response rate of 97.6%; however, the results may differ for very small or very large classes. Additionally, the pandemic-era context during which the course was moved to an online format, should also be taken into account when considering the results of this case. In this case, the course was required to graduate and offered only in the online format. Students who choose a class using this method when given the option between online and in-person sections of the same class may not have the same response. y p The 4.25 hour-long classes were consistent with the regular class length in this degree program. The high-presence online teaching method does not advocate for a particular length of class, but rather suggests adopting the schedule of the program in which a class is taught. This case study details a course taught in a university setting. Undergraduate and master students usually have autonomy and high levels of self-direction. Students meeting with groups in breakout rooms could be relied upon to work on assignments without direct or continuous supervision within the breakout room. Secondary students may have less self-direction and have developed less academic autonomy, which may make this online teaching method more suitable for Honors and AP courses than for all subjects in secondary education. While requiring students to complete an assignment before the first class is plausible in a university setting, there may be difficulties with such a requirement in a secondary education environment. Journal of the Scholarship of Teaching and Learning, Vol. 22, No. 4, December 2022. josotl indiana edu General Lessons The in-person class historically has been held during the same Saturday 8:00 AM - 12:15 PM time slot. This course customarily provides a break about halfway through the class. Several students recommend offering two breaks, providing reasons listed in Table 7. Table 8. Feedback about Class Time Management. If professor give us 2 breaks during the whole class it will be better. The class was 4 hour long. It is a little difficult to sit in front of screen for so long even with breaks. I would like a little bit more break or may be divide into 2 sessions. Maybe one more short break, having only one break after hours when I had my coffee before class didn't feel good always. I think students need more breaks. They don't need to be long, but one 15 minute break for a 4 hour class is simply not enough. I had to get up to go to the bathroom a couple of times and felt my participation grade might have been impacted. Table 8. Feedback about Class Time Management. When offered in-person, the single break is usually about 15 minutes long, however, if students need to use the restroom or answer an important phone call, they can quietly excuse themselves from class for a moment. The webcam-on format seems to discourage similar behavior in the online format. Perhaps a simple announcement at the beginning of the term can alleviate such concerns. One out of the 40 responding students (out of 41 students overall) indicated that they would have preferred the option to attend without using their webcam. That compares to 4 students who expressed that the use of webcams by the whole class was what they most liked about the class. The benefit students get from seeing other students – and the benefit the instructor gets by being able to 45 Gimpel Gimpel see student reactions and look at students when they ask questions or participate in discussions – seems to outweigh reservations about requiring webcam use. see student reactions and look at students when they ask questions or participate in discussions – seems to outweigh reservations about requiring webcam use. Conclusions This case study reports the test of a high-presence, high engagement online teaching method piloted in a master-level process reengineering course. The goal was to capture many of the face-to-face benefits of in-person classes. The course design and instructional technology employed focus on providing interactive immediacy and fostering strong instructor presence. Combined, these contribute to student engagement, which is the most critical factor for student learning and satisfaction with online courses (Martin & Bolliger, 2018; A. 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https://openalex.org/W2159584202	https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0132930&type=printable	English	null	Associations between Food Outlets around Schools and BMI among Primary Students in England: A Cross-Classified Multi-Level Analysis	PloS one	2,015	cc-by	12,450	RESEARCH ARTICLE Abstract Citation: Williams J, Scarborough P, Townsend N, Matthews A, Burgoine T, Mumtaz L, et al. (2015) Associations between Food Outlets around Schools and BMI among Primary Students in England: A Cross-Classified Multi-Level Analysis. PLoS ONE 10(7): e0132930. doi:10.1371/journal.pone.0132930 Methods We measured the densities of fast food outlets and food stores found within schoolchil- dren’s home and school environments using Geographic Information Systems (GIS) and data from local councils. We linked these data to measures from the 2010/11 National Child Measurement Programme and used a cross-classified multi-level approach to examine associations between food retailing and BMI z-scores. Analyses were stratified among Reception (aged 4-5) and Year 6 (aged 10-11) students to measure associations across the primary school years. Data Availability Statement: There are legal restrictions on the student outcome data. National Child Measurement Programme data was used as determined by the data sharing agreement with the then Information Centre of the National Health Service (NHS IC), now Health and Social Care Information Centre (HSCIC). The data sharing agreement stated that all record level data must be treated in accordance with the Data Protection Act 1998 and individual level data must not be made available without agreement of the NHS IC. Cleaned and anonymised NCMP data has subsequently been Introduction Researchers and policy-makers are interested in the influence that food retailing around schools may have on child obesity risk. Most previous research comes from North America, uses data aggregated at the school-level and focuses on associations between fast food outlets and school obesity rates. This study examines associations between food retailing and BMI among a large sample of primary school students in Berkshire, England. By con- trolling for individual, school and home characteristics and stratifying results across the pri- mary school years, we aimed to identify if the food environment around schools had an effect on BMI, independent of socio-economic variables. Editor: Shankuan Zhu, School of Public Health, Zhejiang University, CHINA Received: January 21, 2015 Accepted: June 20, 2015 Published: July 17, 2015 Copyright: © 2015 Williams et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any di id d th i i l th d Editor: Shankuan Zhu, School of Public Health, Zhejiang University, CHINA Editor: Shankuan Zhu, School of Public Health, Zhejiang University, CHINA Editor: Shankuan Zhu, School of Public Health, Zhejiang University, CHINA Received: January 21, 2015 Accepted: June 20, 2015 Published: July 17, 2015 Copyright: © 2015 Williams et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Editor: Shankuan Zhu, School of Public Health, Zhejiang University, CHINA Received: January 21, 2015 Accepted: June 20, 2015 Published: July 17, 2015 Copyright: © 2015 Williams et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Copyright: © 2015 Williams et al. This is an open access article distributed under the terms of the Copyright: © 2015 Williams et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Associations between Food Outlets around Schools and BMI among Primary Students in England: A Cross-Classified Multi-Level Analysis Julianne Williams1, Peter Scarborough1, Nick Townsend1, Anne Matthews1, Thomas Burgoine2, Lorraine Mumtaz1, Mike Rayner1 Julianne Williams1, Peter Scarborough1, Nick Townsend1, Anne Matthews1, Thomas Burgoine2, Lorraine Mumtaz1, Mike Rayner1 a11111 1 British Heart Foundation Centre on Population Approaches for Non-Communicable Disease Prevention, Nuffield Department of Population Health, University of Oxford, Old Road Campus, Oxford, OX3 7LF United Kingdom, 2 UKCRC Centre for Diet and Activity Research (CEDAR), Medical Research Council (MRC) Epidemiology Unit, University of Cambridge School of Clinical Medicine, Institute of Metabolic Science, Cambridge Biomedical Campus, Cambridge, CB2 0QQ United Kingdom * julianne.williams@dph.ox.ac.uk * julianne.williams@dph.ox.ac.uk Introduction Children in England are struggling to meet healthy diet and body weight recommendations [1,2]. According to the most recent estimates, 9.3% of English children in school Reception year (age 4–5 years) and 18.9% in Year 6 (age 10–11 years) are currently obese [3]. Obesity tends to track into adulthood [4] and is difficult to reverse [5,6], which make a strong case for prevention. A growing body of research evidence considers how obesogenic environments [7], which promote energy intake and constrain energy expenditure, may contribute to obesity risk [8– 12]. While research has traditionally focused on environmental exposures near home, there has been increased interest in non-residential environments [13]. Systematic reviews indicate that associations between food retailing and diet [14] or weight status [15] vary across settings and populations and one setting of particular interest is the retail environment around schools. Policy-makers increasingly see school neighbourhoods as a logical place for health promotion as obesity prevention begins in childhood [16–19], schools are well-controlled environments and school-based interventions provide unparalleled access to children because they spend more of their waking hours in school than any other environment outside of home [20]. Competing Interests: The authors have declared that no competing interests exist. Competing Interests: The authors have declared that no competing interests exist. g y A systematic review of studies examining the relationship between food retailing near schools and children’s food purchases, consumption and body weight, found little evidence for an effect of retailing on purchases and consumption, but some evidence of an effect on body weight [2]. However, research in the area is still developing, meaning heterogeneous study designs, methods and measures make it difficult to draw firm conclusions about the effect that retailing near schools may have on a child’s obesity risk [21,22]. The lack of evidence about environmental effects on obesity risk is complicated further by the fact that BMI is the conse- quence of energy balance over time, so the effect of environmental influences may not be immediately apparent. Most of the previous studies have taken place in North America, but food environments and their effects are likely to vary between countries [23], so findings may not be generalisable. Despite these research limitations, planning authorities assume the built environment’s contribution to the obesity epidemic [24–26]. Discussion We found little evidence to suggest that food retailing around schools influences student BMI. There is some evidence to suggest that fast food outlet densities in a child’s home neighbour- hood may have an effect on BMI, particularly among girls, but more research is needed to inform effective policies targeting the effects of the retail environment on child obesity. Funding: This work was undertaken in part by the British Heart Foundation Centre on Population Approaches for Non-Communicable Disease Prevention, a World Health Organization Collaborating Centre and by the Centre for Diet and Activity Research (CEDAR), a UK Clinical Research Collaboration (UKCRC) Public Health Research Centre of Excellence. Funding from the British Heart Foundation (006/P&C/CORE/2013/OXFSTATS, 021/ P&C/Core/2010/HPRG), National Health Services Berkshire project grant, Cancer Research UK, Economic and Social Research Council, Medical Research Council, the National Institute for Health Research, and the Wellcome Trust, under the auspices of the UK Clinical Research Collaboration, is gratefully acknowledged. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Food Outlets around Schools and Student BMI in England positive associations between fast food outlets in home neighbourhood and BMI z-scores. Year 6 students living in areas with the highest density of fast food outlets had an average BMI z-score that was 0.12 (95% CI: 0.04, 0.20) higher than those living in areas with none. positive associations between fast food outlets in home neighbourhood and BMI z-scores. Year 6 students living in areas with the highest density of fast food outlets had an average BMI z-score that was 0.12 (95% CI: 0.04, 0.20) higher than those living in areas with none. made available by the HSCIC through the UK Data Service. Please see below for the contact information of this third party. The food outlet data from the local councils of West Berkshire will be provided in supplemental files. All food outlet data are attributed to their respective local councils: Bracknell Forest Council (http://www.bracknell-forest.gov.uk), Reading Borough Council (http://beta.reading.gov.uk/home), Slough Borough Council (https://www.slough.gov.uk/), The Royal Borough of Windsor and Maidenhead (http://www.rbwm.gov.uk/contacts.htm), Wokingham Borough Council (http://www.wokingham.gov.uk/), West Berkshire Council. To apply for individual-level NCMP data, please contact the Health and Social Care Information Centre, enquiries@hscic.gov.uk. made available by the HSCIC through the UK Data Service. Please see below for the contact information of this third party. The food outlet data from the local councils of West Berkshire will be provided in supplemental files. All food outlet data are attributed to their respective local councils: Bracknell Forest Council (http://www.bracknell-forest.gov.uk), Reading Borough Council (http://beta.reading.gov.uk/home), Slough Borough Council (https://www.slough.gov.uk/), The Royal Borough of Windsor and Maidenhead (http://www.rbwm.gov.uk/contacts.htm), Wokingham Borough Council (http://www.wokingham.gov.uk/), West Berkshire Council. To apply for individual-level NCMP data, please contact the Health and Social Care Information Centre, enquiries@hscic.gov.uk. Results Our multilevel model had three levels to account for individual (n = 16,956), home neighbour- hood (n = 664) and school (n = 268) factors. After controlling for confounders, there were no significant associations between retailing near schools and student BMI, but significant 1 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Food Outlets around Schools and Student BMI in England packaged food [36]. A recent study on energy intakes of US children aged 6 to 11 years found that 63% of energy came from stores, while only 12% came from fast food outlets [37]. Another recent study examined sources of empty calories (the sum of energy from added sugar and solid fat) across retail food stores, schools and fast-food restaurants in the US found that 33% of chil- dren’s empty calories came from stores [38]. In the UK, several pilot studies suggest that food stores on the journey to and from school [39] or near schools [40] may be a major source of cal- ories for school children. Food outlets also tend to cluster around schools [41–43]. In England, longitudinal evidence suggests that numbers of food stores (grocers and convenience stores) and takeaway food outlets in close proximity to schools have increased in recent years [44]. packaged food [36]. A recent study on energy intakes of US children aged 6 to 11 years found that 63% of energy came from stores, while only 12% came from fast food outlets [37]. Another recent study examined sources of empty calories (the sum of energy from added sugar and solid fat) across retail food stores, schools and fast-food restaurants in the US found that 33% of chil- dren’s empty calories came from stores [38]. In the UK, several pilot studies suggest that food stores on the journey to and from school [39] or near schools [40] may be a major source of cal- ories for school children. Food outlets also tend to cluster around schools [41–43]. In England, longitudinal evidence suggests that numbers of food stores (grocers and convenience stores) and takeaway food outlets in close proximity to schools have increased in recent years [44]. In this study we investigated associations between the density of food outlets in both school and home environments and body weight in a large sample of primary school students in Berk- shire, England. We examined if associations varied between types of food outlets. Additionally, we identified if associations were stronger for Reception (ages 4–5) or Year 6 (ages 10–11) stu- dents. Methods Data Individual student characteristics. We used data from the 2010/2011 National Child Measurement Programme (NCMP), which works with local authorities annually to collect data on more than one million children in state-maintained primary schools in England. The NCMP includes measures from 90% of pupils reported eligible to be measured by the primary care trusts. It includes individual height and weight (measured by health professionals, usually school nurses), sex, ethnicity and month of measurement [45]. The NCMP is an anonymized dataset, based around a no consent data collection system. Ethics approval is not required for data col- lection or analysis. As our primary outcome measure, we used body mass index (BMI, kg/m2) to calculate BMI z-scores relative to the International Obesity Task Force reference curves [46]. y School and home characteristics. The NCMP provides each child’s school name and the lower super output area (LSOA) of the home address. LSOAs are a small geographic boundary with a mean population of 1,500 residents [47]. Using data from the Department of Educa- tion’s 2010/2011 census, we ascertained school location, size (total number of pupils), percent- age of students eligible for free school meals and school type (community, foundation, voluntary-controlled or voluntary-aided) [48]. There are different types of maintained or state schools in England: Community schools are controlled by local councils and are not influenced by business or religious groups; foundation schools have more independence to change the way they do things than community schools; voluntary-controlled or voluntary-aided schools are linked to a variety of organisations including faith or charitable organisations [49]. For each child’s home LSOA, we identified the urban/rural classification [50] and child well-being index (CWI). The CWI is a composite score of domains including material well-being, health, education, crime, housing, environment and children in need (information about children who are in various kinds of need and served by local authorities, derived from the Children in Need Survey, from the Department for Children, Schools, and Families) [51]. School and home characteristics. The NCMP provides each child’s school name and the lower super output area (LSOA) of the home address. LSOAs are a small geographic boundary with a mean population of 1,500 residents [47]. Using data from the Department of Educa- tion’s 2010/2011 census, we ascertained school location, size (total number of pupils), percent- age of students eligible for free school meals and school type (community, foundation, voluntary-controlled or voluntary-aided) [48]. PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 We hypothesised that, given that Reception students have not been exposed to the school environment as long as Year 6 students and have less independence than year 6 pupils, the retail outlets around schools are unlikely to have as much of an impact as they would for year 6 students. For this reason, we hypothesised that if the Reception analysis showed null results but the year 6 analyses showed positive results, this may suggest that the food environ- ment around schools has an influence. Introduction In the UK, local authorities have already implemented zoning or licensing restrictions related to hot food takeaway retailing around schools [27], supported by guidelines from the Academy of Medical Royal Colleges [28], Public Health England [29] and the Greater London Authority [30], among others. While the focus in policy discussions [28] and research [31–34] has been on fast-food and takeaway outlets [35], it is important to consider other sources of convenient, energy-dense foods such as grocery stores, convenience stores or petrol stations. A study in New York found that the most frequent sources of food around public schools were small groceries selling mostly 2 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Methods Data There are different types of maintained or state schools in England: Community schools are controlled by local councils and are not influenced by business or religious groups; foundation schools have more independence to change the way they do things than community schools; voluntary-controlled or voluntary-aided schools are linked to a variety of organisations including faith or charitable organisations [49]. For each child’s home LSOA, we identified the urban/rural classification [50] and child well-being index (CWI). The CWI is a composite score of domains including material well-being, health, education, crime, housing, environment and children in need (information about children who are in various kinds of need and served by local authorities, derived from the Children in Need Survey, from the Department for Children, Schools, and Families) [51]. Density of food outlets in school and home environments. We requested food outlet location data from the local councils of Bracknell Forest, Reading, Slough, West Berkshire, 3 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Food Outlets around Schools and Student BMI in England Windsor and Maidenhead, and Wokingham. Environmental health departments of local coun- cils are required to provide this information under the Freedom of Information Act [52]. Food retailers are legally required to register their business with local councils and therefore, this has been found to be a reliable source of food outlet location information in the UK [53,54]. The local councils provided the names and addresses of all individuals, businesses and associations holding a food license. Windsor and Maidenhead, and Wokingham. Environmental health departments of local coun- cils are required to provide this information under the Freedom of Information Act [52]. Food retailers are legally required to register their business with local councils and therefore, this has been found to be a reliable source of food outlet location information in the UK [53,54]. The local councils provided the names and addresses of all individuals, businesses and associations holding a food license. The local council data included records of food outlets that were not of relevance to our research question (for example, industrial food manufacturers or bed and breakfasts), so we established a protocol for cleaning the data and categorizing the food outlet types. Food outlets were grouped into two categories: ‘takeaway and fast food outlets’ or ‘food stores’. Methods Data We defined takeaway and fast food outlets (henceforth referred to as ‘fast food outlets’) as those selling hot or prepared food paid for before consumption [54,55], and which may be consumed off-site, a category that included coffee shops, cafes, pizza shops, sandwich shops, bakeries, delis, kebab shops and takeaway restaurants. We defined food stores as other retail outlets selling food that may be consumed off-site such as supermarkets, convenience stores, off-license stores or news- agents. Researchers consulted local business directories and Google Street View (Mountain View, California, USA) to confirm food outlet types. One researcher completed the initial pro- cessing. To test the reliability of our food outlet definitions, three researchers independently classified a 10% sample of the food outlets and there was agreement for all three raters on 88% of occasions, which gave a kappa score of 0.84. Schools and food outlets were geocoded according to their postcode using a geographic infor- mation system (ArcGIS 10, ESRI, Redlands, CA, USA). Postcodes in the UK contain only 15 addresses on average, and therefore allow for relatively precise geocoding. Postcodes, transpor- tation networks and LSOA boundary data were obtained from Ordnance Survey via UK Digi- map (University of Edinburgh, Scotland, UK). We calculated the density of fast food outlets and food stores located within an 800 metre (m) street network buffer of school centroids and within home LSOA boundaries. Precedent for the use of an 800m buffer to represent a ‘neighbourhood’ has been set in the literature, and approximately corresponds to a 10 minute walk [2]. To test the validity of local council food outlet data, two researchers selected a random 10% sample of schools and identified the food outlets falling within an 800m street network buffer using Google Street View for comparison using percentage agreement statistics. We found that 85% (157 out of 184) of the local council-provided food outlets were also found in Google Street View. However, we also found 22 food outlets in Google Street View that were not included in the local council dataset, so when we compared agreement between the full dataset (using both sources), there was a 76% agreement. Descriptive Statistics We analysed anonymised data from 16,956 individual children attending 268 schools, and living in 664 different home neighbourhoods in Berkshire. Descriptive statistics for the study sample are shown in Table 1. The mean BMI z-score for Reception students (n = 8,745) was 0.38 (SD = 1.00) and for Year 6 students (n = 8,211) was 0.51 (SD = 1.04); these are both lower than the mean BMI z-score found for the national sample in the same calendar year (Reception = 0.46, Year 6 = 0.56). Descriptive statistics for home and school neighbourhood factors, as well as BMI z-scores by home and school neighbourhood characteristics are shown in Table 2. The majority of home neighbourhoods (80.3%) were classified as ‘urban city and town’, and more than half (57.1%) of the participating schools were community schools. Pupils attending schools with a greater proportion of students eligible for free school meals had higher mean BMI z-scores. Food outlet frequencies within home and school neighbourhoods are shown in Table 3. The number of fast food outlets found within home neighbourhoods ranged from 0 to 35, with a mean of 1.14 (SD = 3.05). The number of food stores ranged from 0 to 35, with a mean of 1.40 (SD = 3.03). There were more fast food outlets and food stores in the most deprived home neighbourhoods (i.e. those falling within the highest quartiles of the Child Well Being Index). The number of fast food outlets found within an 800 metre street network of schools ranged from 0 to 30 outlets, with a mean of 2.67 (SD = 4.47). The number of food stores ranged from 0 to 33, with a mean of 3.34 (SD = 4.57). There were more fast food outlets and food stores located near larger schools and those with the highest proportion of students eligible for free school meals (Table 3). A description of participant BMI z-scores by home and school neighbourhoods is shown in Table 4. Reception students living in LSOAs with no fast food outlets had a mean BMI z-score of 0.36 (SD = 0.33), while those living in LSOAs with 30–35 fast food outlets had a BMI z score of 0.90 (SD = 0.63). Food Outlets around Schools and Student BMI in England Three-level cross-classified random effects models were run using Markov chain Monte Carlo methods. Four models were calculated for each year group: Model 1 (the null model) was run for each exposure variable only (fast food near schools, food stores near schools, fast food in home neighbourhood and food stores in home neighbourhood) and controlled for no covar- iates, this allowed us to examine any associations between BMI z-score and food density at schools and home in univariable models. Model 2 included home and school neighbourhood covariates, and were run for each outcome variable separately, allowing us to study if any asso- ciations in Model 1 were due to confounding factors at the school or home neighbourhood level. Model 3 was also run for each outcome variable and controlled for all covariates includ- ing those at the school, home and individual levels, to investigate if any significant associations could be explained by differences between schools and home neighbourhoods in terms of pupil composition. The full model 4 included the school, home and individual level covariates, and all outcome variables together; this final model investigated whether food store densities around schools and homes had independent associations with BMI z-score. Ninety five percent confidence intervals (CIs) were used to determine significant differences between tertiles of outlet density and BMI z-score. We also used Wald tests to determine whether the non-refer- ence categories were significantly different to each other in their association with the outcome variable. All analyses were conducted using MLwiN version 2.28 (Centre for Multilevel Model- ling, University of Bristol, UK). Statistical analysis Using a cross-classified multi-level model, which allowed us to account for the nested structure of the data (i.e. children within school and home neighbourhoods), we examined the associa- tion between pupil BMI z-score and density of fast food and food stores in both home and school areas while controlling for confounding factors at the individual student, school and home environment levels. We categorised food outlet densities into ‘0 outlets’ (the reference category) and divided the remaining densities into tertiles. The individual-level student vari- ables were sex, age, ethnicity and month of measurement. Home neighbourhood-level variables were urbanicity and residential child well-being index. School neighbourhood-level variables were school size, type and percentage of students on free meals. The results for BMI are reported as z-scores in comparison to the International Obesity Task Force standard [56]. 4 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Food Outlets around Schools and Student BMI in England Table 1. Frequencies and mean BMI z-scores for individual-level factors. Descriptive statistics Reception Year 6 Total BMI z-score Reception Year 6 Mean (SD) Mean (SD) Individual- n 8,745 8,211 16,956 All 0.38 (1.00) 0.51 (1.04) Sex—(%) Sex Male 4,501 (51.5) 4,265 (51.9) 8,766 (51.7) Male 0.34 (1.00) 0.49 (1.05) Female 4,244 (48.5) 3,946 (48.1) 8,190 (48.3) Female 0.42 (1.00) 0.52 (1.04) Age in months Age quartiles Mean (SD) 63.0 (5.3) 133.4 (3.8) 97.1 (35.5) 1st Quartile 0.43 (0.98) 0.51 (1.04) Min-Max 48.9–77.6 124.5–142.3 48.9–142.3 2nd Quartile 0.42 (0.98) 0.49 (1.03) 3rd Quartile 0.34 (0.99) 0.52 (1.06) 4th Quartile 0.32 (1.03) 0.51 (1.04) Ethnicity—(%) Ethnicity Bangladeshi 14 (0.2) 42 (0.5) 56 (0.3) Bangladeshi 0.02 (1.16) 1.01 (1.24) Black African 90 (1.0) 228 (2.8) 318 (1.9) Black African 0.39 (1.19) 0.76 (1.04) Black Caribbean 25 (0.3) 75 (0.9) 100 (0.6) Black Caribbean -0.26 (0.84) 0.66 (1.24) Chinese 10 (0.1) 49 (0.6) 59 (0.3) Chinese 0.21 (0.55) 0.22 (1.12) Indian 195 (2.2) 410 (5.0) 605 (3.6) Indian 0.10 (1.16) 0.58 (1.15) Pakistani 363 (4.2) 591 (7.2) 954 (5.6) Pakistani 0.08 (1.16) 0.57 (1.27) White British 1,789 (20.5) 5,520 (67.2) 7,309 (43.1) White British 0.39 (0.89) 0.46 (0.99) White Irish 19 (0.2) 44 (0.5) 63 (0.4) White Irish 0.58 (0.94) 0.67 (0.88) Other 398 (4.6) 1,169 (14.2) 1,567 (9.2) Other 0.35 (1.11) 0.62 (1.07) Not stated 5,842 (66.8) 83 (1.0) 5,925 (34.9) Not stated 0.41 (0.99) 0.39 (1.09) Month measured—n (%) Month measured Sept—Dec 2,532 (29.0) 0 (0) 2,532 (14.9) Sept—Dec 0.40 (1.02) - Jan—April 4,313 (49.3) 4,851 (59.1) 9,164 (54.0) Jan—April 0.38 (0.98) 0.49 (1.03) May—Aug 1,900 (21.7) 3,360 (40.9) 5,260 (31.0) May—Aug 0.34 (0.99) 0.54 (1.06) doi:10 1371/journal pone 0132930 t001 Table 1. Frequencies and mean BMI z-scores for individual-level factors. Reception and Year 6 students attending schools with no fast food or no food stores within 800 metres had lower mean BMI z-scores than those attending schools with the highest densities of food outlets. Descriptive Statistics Among Year 6 students, there was relatively less of a difference between pupils living in areas with no food outlets compared to those living in areas with the highest density of outlets, with BMI z scores of 0.51 (SD = 0.45) and 0.45 (SD = 0.43), respectively. 5 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Food Outlets around Schools and Student BMI in England Table 2. Frequencies and mean BMI z-scores for home- and school-level factors. Table 2. Frequencies and mean BMI z-scores for home- and school-level factors. Home Lower Super Output Area (LSOA) Factors Descriptive Statistics Reception Year 6 Total BMI z-score Reception Year 6 LSOA—n 614 628 664 Mean (SD) Mean (SD) Urban/rural—(%) Urban/rural Rural town and fringe 52 (8.5) 52 (8.3) 57 (8.6) Rural town and fringe 0.42 (0.33) 0.54 (0.56) Rural village and dispersed 43 (7.0) 45 (7.2) 51 (7.7) Rural village and dispersed 0.37 (0.36) 0.22 (0.57) Urban city and town 509 (82.9) 516 (82.2) 533 (80.3) Urban city and town 0.36 (0.33) 0.53 (0.41) Urban major conurbation 10 (1.6) 15 (2.4) 23 (3.5) Urban major conurbation 0.72 (0.89) 0.64 (1.03) Child Well Being Index Child Well Being Index Mean (SD) 100.4 (73.5) 99.5 (73.3) 99.5 (72.79) 1st Quartile 0.33 (0.36) 0.45 (0.48) Min-Max 9.2–400.8 9.2–400.8 9.2–400.8 2nd Quartile 0.30 (0.33) 0.46 (0.47) 3rd Quartile 0.44 (0.36) 0.50 (0.49) 4th Quartile 0.43 (0.32) 0.61 (0.47) School-level factors Descriptive Statistics Reception Year 6 Total BMI z-score Reception Year 6 School-n 238 221 268 Mean (SD) Mean (SD) School type—(%) School type Community 135 (56.7) 125 (56.6) 153 (57.1) Community 0.38 (0.21) 0.51 (0.22) Foundation 6 (2.5) 6 (2.7) 6 (2.2) Foundation 0.38 (0.32) 0.72 (0.28) Voluntary aided 55 (23.1) 54 (24.4) 62 (23.1) Voluntary aided 0.36 (0.23) 0.47 (0.28) Voluntary controlled 42 (17.6) 36 (16.3) 47 (17.5) Voluntary controlled 0.41 (0.23) 0.50 (0.31) Total no. Pupils—(%) Total no. Pupils Mean (SD) 268.9 (150.0) 285.4 (150.6) 276.8 (144.3) 1st Quartile 0.38 (0.27) 0.44 (0.31) Min-Max 29.0–763.0 29.0–763.0 29.0–763.0 2nd Quartile 0.38 (0.22) 0.49 (0.24) 3rd Quartile 0.41 (0.21) 0.56 (0.20) 4th Quartile 0.37 (0.21) 0.53 (0.23) Free School Meals—n (%) Free School Meals Mean (SD) 10.1 (8.8) 10.5 (9.4) 10.1 (9.0) 1st Quartile 0.30 (0.20) 0.37 (0.21) Min-Max 0.0–46.7 0.0–48.2 0.0–48.2 2nd Quartile 0.40 (0.21) 0.47 (0.22) 3rd Quartile 0.41 (0.24) 0.57 (0.26) 4th Quartile 0.42 (0.23) 0.61 (0.24) Home Lower Super Output Area (LSOA) Factors Home Lower Super Output Area (LSOA) Factors doi:10.1371/journal.pone.0132930.t002 the analyses by sex, we found that the negative relationship between food store densities and BMI was significant for boys but not girls (mean BMI z-score = -0.09, 95% CI: -0.18, -0.01). There was a positive relationship between BMI z-scores and home fast food outlet density for Year 6 students. Those who were exposed to the highest densities of fast food outlets in the home neighbourhood had BMI z-scores 0.10 (95% CI: 0.01, 0.18) and 0.12 (95% CI: 0.04, 0.20) greater than those least exposed. When we stratified the analyses by sex, we found that the rela- tionship between fast food outlets near homes and BMI z-score was significant for girls, but not for boys. Year 6 girls living areas with the highest densities of fast food had a mean BMI z- score that was 0.19 (95% CI: 0.07, 0.3) greater than those living in areas with none. Adjusted associations between BMI z-scores and school neighbourhood fast food or food store densities were null for Year 6 students. Associations between the density of food outlets and BMI z-scores School neighbourhoods. Results from the cross-classified multilevel analysis are shown in Tables 5 and 6. For Reception students overall, there were no significant associations between school fast food or food store densities and BMI (Table 5). For Year 6 students, initial models showed a significant positive relationship between school fast food and food store densities and BMI z-score, however these associations were null when adjusting for school, home or individ- ual-level covariates (Table 6). Home neighbourhoods. In model 4, Reception students living in home neighbourhoods with the highest densities of fast food had a mean BMI z-score 0.08 (95% CI: 0.00, 0.17) higher than those living in home neighbourhoods with no fast food stores. When we stratified the analyses by sex, we found that this relationship was significant for Reception girls (mean BMI z-score = 0.13, 95% CI 0.00, 0.24), but not boys. However, evidence of a dose-response effect in this relationship was not observed. Reception students whose home neighbourhoods had the highest density of food stores had a mean BMI z-score that was 0.09 (95% CI: -0.16, -0.01) lower than students living in home neighbourhoods with no food stores, however associations with other densities of home neighbourhood food store exposure were null. When we stratified PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 6 / 17 Discussion This analysis did not support an independent effect of food stores or fast food outlets around schools on body weight in a sample of UK Reception or Year 6 students. However, there was PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 7 / 17 Food Outlets around Schools and Student BMI in England Table 3. Food outlet frequencies by home and school neighbourhoods. Food outlets within home Lower Super Output Area (LSOA) Store Frequency Fast food (n = 630) Food stores (n = 630) Mean (SD) Mean (SD) 1.14 (3.05) 1.40 (3.03) Child Well Being Index 1st Quartile 0.74 (2.95) 1.05 (2.74) 2nd Quartile 0.88 (2.17) 1.14 (2.20) 3rd Quartile 1.50 (4.03) 1.72 (2.67) 4th Quartile 1.49 (2.72) 1.72 (2.66) Urban/rural Rural town and fringe 0.70 (1.60) 1.09 (1.71) Rural village and dispersed 0.57 (1.41) 0.88 (1.61) Urban city and town 1.27 (3.30) 1.52 (3.26) Urban major conurbation 0 (0) 0 (0) Food outlets near schools (within 800 metres) Store Frequency Fast food (n = 712) Food stores (n = 892) Mean (SD) Mean (SD) 2.67 (4.47) 3.34 (4.57) School type Community 2.8 (4.4) 3.5 (4.8) Foundation 3.8 (3.4) 5.7 (5.1) Voluntary aided 2.8 (5.2) 3.4 (4.8) Voluntary controlled 1.8 (3.6) 2.5 (3.1) Total no. Pupils 1st Quartile 1.0 (2.9) 1.7 (3.4) 2nd Quartile 2.0 (3.0) 2.8 (3.2) 3rd Quartile 4.3 (6.0) 4.8 (6.2) 4th Quartile 3.5 (2.7) 4.1 (4.2) Free Meals 1st Quartile 1.5 (3.6) 2.0 (3.8) 2nd Quartile 1.6 (2.6) 2.2 (3.0) 3rd Quartile 3.2 (4.9) 3.9 (4.3) 4th Quartile 4.3 (5.6) 5.2 (5.9) doi:10.1371/journal.pone.0132930.t003 Table 3. Food outlet frequencies by home and school neighbourhoods. Table 3. Food outlet frequencies by home and school neighbourhoods. doi:10.1371/journal.pone.0132930.t003 evidence of a weak positive association between fast food outlet exposure around the home and body weight for older girls in this sample, which remained after adjustment for individual-level and school-level covariates, and exposure to other types of food stores. There are a number of strengths and limitations to this paper. One of the main strengths was our use of school, home and individual-level measures, which enabled us to identify multi-level determinants of body weight while accounting for the cross-classified structure of the data [57]. Food Outlets around Schools and Student BMI in England Table 4. BMI z-scores by home and school neighbourhoods. Table 4. BMI z-scores by home and school neighbourhoods. Food outlets within home Lower Super Output Areas (LSOAs) BMI z-score BMI z-score Reception Year 6 Reception Year 6 N-LSOAs % Mean (SD) Mean (SD) N-LSOAs % Mean (SD) Mean (SD) Fast food 0.37 (0.33) 0.51 (0.45) Food stores 0.37 (0.34) 0.51 (0.45) 0 404 64.1 0.36 (0.37) 0.49 (0.50) 0 320 50.8 0.37 (0.39) 0.51 (0.54) 1 88 14 0.37 (0.28) 0.47 (0.340 1 131 20.8 0.36 (0.28) 0.48 (0.36) 2 52 8.3 0.44 (0.29) 0.61 (0.32) 2 75 11.9 0.39 (0.28) 0.56 (0.34) 3 32 5.1 0.38 (0.28) 0.55 (0.42) 3 37 5.9 0.34 (0.30) 0.48 (0.35) 4 16 2.5 0.32 (0.30) 0.63 (0.37) 4 26 4.1 0.39 (0.30) 0.60 (0.36) 5 14 2.2 0.33 (0.18) 0.62 (0.38) 5 16 2.5 0.30 (0.29) 0.60 (0.50) 6–9 15 2.4 0.54 (0.20) 0.69 (0.31) 6–9 16 2.5 0.45 (0.28) 0.53 (0.35) 10–19 6 1 0.43 (0.22) 0.49 (0.19) 10–19 4 0.6 0.45 (0.28) 0.38 (0.16) 20–29 0 0 - - 20–29 3 0.5 0.36 (0.22) 0.52 (0.7) 30–35 2 0.3 0.90 (0.63) 0.45 (0.43) 30–35 2 0.3 0.42 (0.07) 0.39 (0.60) Food outlets near schools (within 800 metres) BMI z-score BMI z-score Reception Year 6 Reception Year 6 N-schools % Mean (SD) Mean (SD) N-schools % Mean (SD) Mean (SD) Fast food 0.38 (0.23) 0.51 (0.25) Food stores 0.38 (0.23) 0.51 (0.25) 0 119 44.6 0.38 (0.24) 0.45 (0.25) 0 74 27.7 0.35 (0.23) 0.40 (0.20) 1 34 12.7 0.36 (0.24) 0.52 (0.29) 1 46 17.2 0.39 (0.22) 0.52 (0.30) 2 35 13.1 0.36 (0.18) 0.56 (0.20) 2 40 15 0.42 (0.26) 0.54 (0.24) 3 20 7.5 0.32 (0.21) 0.50 (0.25) 3 25 9.4 0.39 (0.19) 0.50 (0.18) 4 12 4.5 0.45 (0.27) 0.59 (0.19) 4 21 7.9 0.45 (0.20) 0.65 (0.27) 5 5 1.9 0.33 (0.18) 0.62 (0.08) 5 10 3.7 0.26 (0.14) 0.46 (0.25) 6–9 19 7.1 0.42 (0.15) 0.47 (0.22) 6–9 26 9.7 0.42 (0.22) 0.62 (0.23) 10–19 19 7.1 0.50 (0.24) 0.73 (0.23) 10–19 22 8.2 0.37 (0.26) 0.55 (0.24) 20–29 3 1.1 0.41 (0.09) 0.68 (0.27) 20–29 2 0.7 0.47 (0.05) 0.57 (-) 30 - 0.19 (-) 30–35 1 0.4 0.38 (-) 0.98 (-) doi:10.1371/journal.pone.0132930.t004 Table 4. BMI z-scores by home and school neighbourhoods. of environmental exposure based around key daily anchor points such as homes and schools, as well as the potential moderating influence of individual-level characteristics such as gender. of environmental exposure based around key daily anchor points such as homes and schools, as well as the potential moderating influence of individual-level characteristics such as gender. Few studies have reported on the reliability or validity of their methods to characterize food environments. We conducted inter-rater reliability tests for our food outlet inclusion criteria and categorization, finding 88% agreement, which gave a kappa score of 0.84. We also validated the data provided from the local councils against Google Street View, a virtual street audit method that has been identified as a potentially promising alternative to ground-truthing [59]. We found a low level of agreement between the two sources, which may have been due to temporal mis- match between local council records and Google’s images. Earlier attempts to validate local council food outlet data with Google Street View found similarly low levels of agreement [60]. The acceptability and validity of secondary food outlet data sources, including data from local councils in the United Kingdom, has been tested previously against field observations and has been found to be the most accurate publicly available source of food environment data compared to the ‘gold standard’ of field testing[52]. Lake et al compared three sources of food environment data (Yellow Pages, Yell.com and Council Data) to a ‘gold standard’ of observa- tions made in the field and found that the council data had a positive predictive value of 91.5% Few studies have reported on the reliability or validity of their methods to characterize food environments. We conducted inter-rater reliability tests for our food outlet inclusion criteria and categorization, finding 88% agreement, which gave a kappa score of 0.84. We also validated the data provided from the local councils against Google Street View, a virtual street audit method that has been identified as a potentially promising alternative to ground-truthing [59]. We found a low level of agreement between the two sources, which may have been due to temporal mis- match between local council records and Google’s images. Earlier attempts to validate local council food outlet data with Google Street View found similarly low levels of agreement [60]. Discussion The importance of accounting for individual characteristics and exposures from multiple loca- tions is highlighted by Kestens et al, who found that estimates of food environment exposures accounting for both residential and non-residential settings were significantly and more strongly associated with overweight than estimates based on one exposure setting only [58]. This rela- tionship differed between men and women, highlighting the importance of multiple estimates PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 8 / 17 Food outlets within home Lower Super Output Areas (LSOAs) BMI z-score BMI z-score Reception Year 6 Reception Year 6 N-LSOAs % Mean (SD) Mean (SD) N-LSOAs % Mean (SD) Mean (SD) Fast food 0.37 (0.33) 0.51 (0.45) Food stores 0.37 (0.34) 0.51 (0.45) 0 404 64.1 0.36 (0.37) 0.49 (0.50) 0 320 50.8 0.37 (0.39) 0.51 (0.54) 1 88 14 0.37 (0.28) 0.47 (0.340 1 131 20.8 0.36 (0.28) 0.48 (0.36) 2 52 8.3 0.44 (0.29) 0.61 (0.32) 2 75 11.9 0.39 (0.28) 0.56 (0.34) 3 32 5.1 0.38 (0.28) 0.55 (0.42) 3 37 5.9 0.34 (0.30) 0.48 (0.35) 4 16 2.5 0.32 (0.30) 0.63 (0.37) 4 26 4.1 0.39 (0.30) 0.60 (0.36) 5 14 2.2 0.33 (0.18) 0.62 (0.38) 5 16 2.5 0.30 (0.29) 0.60 (0.50) 6–9 15 2.4 0.54 (0.20) 0.69 (0.31) 6–9 16 2.5 0.45 (0.28) 0.53 (0.35) 10–19 6 1 0.43 (0.22) 0.49 (0.19) 10–19 4 0.6 0.45 (0.28) 0.38 (0.16) 20–29 0 0 - - 20–29 3 0.5 0.36 (0.22) 0.52 (0.7) 30–35 2 0.3 0.90 (0.63) 0.45 (0.43) 30–35 2 0.3 0.42 (0.07) 0.39 (0.60) Food outlets near schools (within 800 metres) Food outlets within home Lower Super Output Areas (LSOAs) BMI z-score BMI z-score BMI z-score Reception Year 6 Reception Year 6 N-schools % Mean (SD) Mean (SD) N-schools % Mean (SD) Mean (SD) Fast food 0.38 (0.23) 0.51 (0.25) Food stores 0.38 (0.23) 0.51 (0.25) 0 119 44.6 0.38 (0.24) 0.45 (0.25) 0 74 27.7 0.35 (0.23) 0.40 (0.20) 1 34 12.7 0.36 (0.24) 0.52 (0.29) 1 46 17.2 0.39 (0.22) 0.52 (0.30) 2 35 13.1 0.36 (0.18) 0.56 (0.20) 2 40 15 0.42 (0.26) 0.54 (0.24) 3 20 7.5 0.32 (0.21) 0.50 (0.25) 3 25 9.4 0.39 (0.19) 0.50 (0.18) 4 12 4.5 0.45 (0.27) 0.59 (0.19) 4 21 7.9 0.45 (0.20) 0.65 (0.27) 5 5 1.9 0.33 (0.18) 0.62 (0.08) 5 10 3.7 0.26 (0.14) 0.46 (0.25) 6–9 19 7.1 0.42 (0.15) 0.47 (0.22) 6–9 26 9.7 0.42 (0.22) 0.62 (0.23) 10–19 19 7.1 0.50 (0.24) 0.73 (0.23) 10–19 22 8.2 0.37 (0.26) 0.55 (0.24) 20–29 3 1.1 0.41 (0.09) 0.68 (0.27) 20–29 2 0.7 0.47 (0.05) 0.57 (-) 30 - 0.19 (-) 30–35 1 0.4 0.38 (-) 0.98 (-) doi:10.1371/journal.pone.0132930.t004 of environmental exposure based around key daily anchor points such as homes and schools, as well as the potential moderating influence of individual-level characteristics such as gender. doi:10.1371/journal.pone.0132930.t004 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 Food Outlets around Schools and Student BMI in England Table 5. Associations between food outlets in home Lower Super Output Area (LSOA) and within 800 metres of schools and BMI z-score among Reception students in Berkshire, England. Food Reception Students Overall Boys Girls Outletsa Model 1 Model 2 Model 3 Model 4 Model 4 Model 4 ref = 0 β 95% CI β 95% CI β 95% CI β 95% CI β 95% CI β 95% CI Fast food around schools (ref = 0) 1 -0.011 -0.08, 0.058 -0.01 -0.08, 0.061 0.005 -0.064, 0.073 -0.01 -0.086, 0.065 0.004 -0.094, 0.103 -0.028 -0.127, 0.071 2 0.019 -0.068, 0.107 0.025 -0.063, 0.113 0.041 -0.046, 0.128 0.046 -0.056, 0.147 0.055 -0.071, 0.182 0.037 -0.088, 0.163 3 0.059 -0.015, 0.133 0.048 -0.032, 0.128 0.058 -0.019, 0.135 0.113 0,0.226 0.121 -0.020, 0.263 0.087 -0.056, 0.231 Wald 2.949 (p = 0.229) 1.957 (p = 0.376) 1.854 (p = 0.396) 5.405 (p = 0.067) 2.979 0.226 2.974 0.226 Food stores around schools (ref = 0) 1 0.04 -0.034, 0.114 0.038 -0.039, 0.115 0.043 -0.031, 0.118 0.045 -0.034, 0.125 0.025 -0.077, 0.128 0.064 -0.037, 0.166 2 0.077 -0.008, 0.162 0.058 -0.028, 0.144 0.055 -0.03, 0.141 0.032 -0.07, 0.135 0.038 -0.093, 0.168 0.034 -0.098, 0.167 3 0.022 -0.056, 0.1 -0.006 -0.094, 0.083 0.029 -0.056, 0.113 -0.05 -0.173, 0.074 -0.069 -0.223, 0.086 -0.020 -0.179, 0.139 Wald 3.392 (p = 0.335 2.333 (p = 0.311) 0.411 (p = 0.814) 3.453 (p = 0.178) 3.056 0.217 1.538 0.464 Fast food in home Lower Super Output Area (ref = 0) 1 -0.012 -0.075, 0.051 -0.011 -0.073, 0.051 -0.006 -0.068, 0.056 0.015 -0.05, 0.08 0.005 -0.087, 0.097 0.022 -0.073, 0.117 2 0.067 -0.011, 0.145 0.06 -0.016, 0.135 0.058 -0.019, 0.134 0.084* 0.001, 0.167 0.043 -0.074, 0.161 0.125* 0.006, 0.244* 3 0.008 -0.051, 0.067 -0.005 -0.065, 0.055 -0.001 -0.058, 0.056 0.042 -0.031, 0.115 0.045 -0.062, 0.152 0.047 -0.057, 0.151 Wald 3.367 (p = 0.339) 2.777 (p = 0.249) 2.289 (p = 0.318) 2.286 (p = 0.319) 0.568 0.753 2.530 0.282 Food stores in home Lower Super Output Area (ref = 0) 1 -0.02 -0.079, 0.038 -0.03 -0.088, 0.028 -0.027 -0.085, 0.031 -0.045 -0.105, 0.014 -0.092 -0.176, -0.007* -0.002 -0.087, 0.083 2 -0.006 -0.073, 0.062 -0.008 -0.073, 0.058 -0.006 -0.073, 0.06 -0.037 -0.11, 0.036 -0.012 -0.112, 0.089 -0.057 -0.161, 0.048 3 -0.034 -0.094, 0.025 -0.055 -0.115, 0.006 -0.048 -0.106, 0.01 -0.086* -0.162,- 0.011 -0.096 -0.202, 0.011 -0.082 -0.189, 0.026 Wald 0.588 (p = 0.745) 1.601 (p = 0.449) 1.280 (p = 0.527) 1.689 (p = 0.43) 3.034 0.219 2.384 0.304 Model 1 Univariable models for each exposure variable, Model 2 Univariable exposure variables with controls for school (number of pupils, percentage eligible for free meals, school type, school urbanicity) and LSOA (urbanicity and IMD), Model 3 Multivariable- Univariable exposure with all controls (school, LSOA and pupil: sex age, month of measurement, ethnicity), Model 4 Multivariable exposure with all controls. h b l d l d f d f d l d l d d f The acceptability and validity of secondary food outlet data sources, including data from local councils in the United Kingdom, has been tested previously against field observations and has been found to be the most accurate publicly available source of food environment data compared to the ‘gold standard’ of field testing[52]. Lake et al compared three sources of food environment data (Yellow Pages, Yell.com and Council Data) to a ‘gold standard’ of observa- tions made in the field and found that the council data had a positive predictive value of 91.5% PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 9 / 17 doi:10.1371/journal.pone.0132930.t005 Food Outlets around Schools and Student BMI in England Table 6. Associations between food outlets in home Lower Super Output Area (LSOA) and within 800 metres of schools and BMI z-score among Year 6 students in Berkshire, England. Food Year 6 Students Overall Boys Girls Outletsa Model 1 Model 2 Model 3 Model 4 Model 4 Model 4 ref = 0 β 95% CI β 95% CI β 95% CI β 95% CI β 95% CI β 95% CI Fast food around schools (ref = 0) 1 0.064 -0.012, 0.14 0.016 -0.058, 0.09 0.029 -0.05, 0.108 -0.008 -0.089, 0.073 0.002 -0.096, 0.100 -0.001 -0.110, 0.108 2 0.059 -0.039, 0.157 0.009 -0.083, 0.101 -0.002 -0.096, 0.092 -0.033 -0.142, 0.077 -0.002 -0.133, 0.129 -0.019 -0.165, 0.126 3 0.121* 0.037, 0.205 0.039 -0.041, 0.119 0.028 -0.059, 0.116 0.038 -0.094, 0.17 0.101 -0.052, 0.254 0.018 -0.152, 0.187 Wald 1.780 (p = 0.411) 0.440 (p = 0.803) 0.485 (p = 0.785) 1.546 (p = 0.462) 2.511 p = -0.285 0.241 p = -0.886 Food stores around schools (ref = 0) 1 0.121* 0.041, 0.201 0.059 -0.019, 0.137 0.073 -0.01, 0.155 0.085 -0.003, 0.174 0.034 -0.070, 0.138 0.113 0.000, 0.227 2 0.148* 0.052, 0.244 0.064 -0.032, 0.16 0.075 -0.021, 0.172 0.096 -0.017, 0.21 0.056 -0.077, 0.189 0.106 -0.043, 0.254 3 0.161* 0.077, 0.245 0.037 -0.045, 0.119 0.036 -0.051, 0.123 0.018 -0.124, 0.159 -0.053 -0.212, 0.106 0.070 -0.110, 0.250 Wald 1.109* (p = 0.574) 0.486 (p = 0.784) 1.128 (p = 0.569) 2.288 (p = 0.319) 2.744 p = -0.254 0.297 p = -0.862 Fast food in home Lower Super Output Area (ref = 0) 1 0.009 -0.062, 0.08 0.013 -0.054, 0.08 0.013 -0.057, 0.084 0.024 -0.044, 0.092 -0.029 -0.127, 0.069 0.058 -0.044, 0.160 2 0.106* 0.018, 0.194 0.089 0.003, 0.175 0.079 -0.008, 0.166 0.096* 0.011, 0.181 -0.046 -0.171, 0.079 0.246* 0.115, 0.377 3 0.117* 0.052, 0.182 0.086 0.017, 0.155 0.083* 0.018, 0.147 0.120* 0.042, 0.198 0.041 -0.071, 0.153 0.185* 0.067, 0.303 Wald 6.879* (p = 0.032) 3.519 (p = 0.172) 3.072 (p = 0.215) 4.448 (p = 0.108) 2.024 p = -0.363 6.119* p = -0.047 Food stores in home Lower Super Output Area (ref = 0) 1 -0.014 -0.083, 0.055 -0.028 -0.091, 0.035 -0.028 -0.088, 0.032 -0.046 -0.108, 0.017 -0.025 -0.111, 0.061 -0.085 -0.174, 0.005 2 0.087* 0.005, 0.169 0.084* 0.01, 0.158* 0.079* 0.008, 0.151 0.032 -0.05, 0.114 0.107 -0.007, 0.221 -0.073 -0.191, 0.044 3 0.049 -0.02, 0.118 0.012 -0.053, 0.077 0.004 -0.062, 0.069 -0.073 -0.154, 0.008 0.013 -0.101, 0.127 -0.165 -0.283,- 0.046 Wald 5.888 (p = 0.527) 7.731* (p = 0.021) 6.517* (p = 0.038) 5.331 (p = 0.070) 5.361 p = -0.069 2.238 p = -0.327 Model 1 Univariable models for each exposure variable, Model 2 Univariable exposure variables with controls for school (number of pupils, percentage eligible for free meals, school type, school urbanicity) and LSOA (urbanicity and IMD), Model 3 Multivariable- Univariable exposure with all controls (school, LSOA and pupil: sex age, month of measurement, ethnicity), Model 4 Multivariable exposure with all controls. aFood outlet densities were categorised into zero outlets (the reference group) and the remaining densities were divided into tertiles. Indicates signiﬁcant associations (p <0.05) tlets in home Lower Super Output Area (LSOA) and within 800 metres of schools and BMI z-score among nd Model 1 Univariable models for each exposure variable, Model 2 Univariable exposure variables with controls for school (number of pupils, percentage eligible for free meals, school type, school urbanicity) and LSOA (urbanicity and IMD), Model 3 Multivariable- Univariable exposure with all controls (school, LSOA and pupil: sex age, month of measurement, ethnicity), Model 4 Multivariable exposure with all controls. aFood outlet densities were categorised into zero outlets (the reference group) and the remaining densities were divided into tertiles. Indicates signiﬁcant associations (p <0.05) doi:10.1371/journal.pone.0132930.t005 [54]. Similarly, Cummins et al [53] found reasonable, but imperfect agreement between a pub- licly available directory of food retail premises and field-validated reality. However, while coun- cil data is likely to be in the upper bounds of validity compared to commercial listings (because it is collected for regulatory purposes), researchers should acknowledge the imperfect nature of such data [53]. Our study was limited by its cross-sectional design, which precluded us from establishing a causal relationship between food outlet exposure and body weight. While the NCMP data 10 / 17 PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 doi:10.1371/journal.pone.0132930.t006 Food Outlets around Schools and Student BMI in England Questions remain about defining and classifying food outlets. In this study, we drew upon studies by Lake et al[54] and Burgoine et al [55] in forming our definitions of fast food outlets and food stores. However, definitions vary widely between studies. A review on fast food access by Fleischacker et al [64] found that close to half of the studies (n = 16, 40%) used a proprietary fast food outlet definition. Given this variation, it is important for future research to be clear about their criteria for classifying food outlets. Food access includes many components [14] which were beyond the scope of this study to measure. Our measures of food availability (density of fast food outlets and food stores) may not reflect the foods actually available within stores. Categorizing food outlets as ‘healthy’ (e.g. grocery stores) and ‘unhealthy’ (e.g. takeaway shops) is problematic [65] because both healthy and unhealthy food options are often found within the same venue. Future research needs to integrate detailed information on what is sold within stores, alongside measures of access to the stores themselves [22]. However, it is possible that use of food stores varies as a function of age such that young children make less healthy choices than older children and adults. This hypothesis is yet to be explored in the published literature. This study did not consider the food environments within schools, which have broad poten- tial to impact pupil’s food choices [66–68]. School meal standards in the United Kingdom have been described as amongst some of the most detailed and comprehensive in the world[69] and recent reforms from the Department of Education have introduced a new set of standards for all food served in schools [70]. In contexts where the within-school food environment is less regulated (or where standards are non-existent), future research should consider both the within-school and out-of-school availability of food when considering environmental influ- ences on diet quality. The NCMP data allowed us to control for individual-level characteristics such as sex, age, and ethnicity, but it did not include other potentially important factors, such as cognitions, psychological and psychosocial factors [71]. Our school data set would have been strengthened by additional measures on socio-cultural and political environments [72]. aFood outlet densities were categorised into zero outlets (the reference group) and the remaining densities were divided into tertiles. Indicates signiﬁcant associations (p <0.05) tlets in home Lower Super Output Area (LSOA) and within 800 metres of schools and BMI z-score among Table 6. Associations between food outlets in home Lower Super Output Area (LSOA) and within 800 metre Year 6 students in Berkshire, England. Model 1 Univariable models for each exposure variable, Model 2 Univariable exposure variables with controls for school (number of pupils, percentage eligible for free meals, school type, school urbanicity) and LSOA (urbanicity and IMD), Model 3 Multivariable- Univariable exposure with all controls (school, LSOA and pupil: sex age, month of measurement, ethnicity), Model 4 Multivariable exposure with all controls. aFood outlet densities were categorised into zero outlets (the reference group) and the remaining densities were divided into tertiles. Indicates signiﬁcant associations (p <0.05) doi:10.1371/journal.pone.0132930.t006 provided BMI measures for a large number of children, it did not include other data on physi- cal activity or dietary behaviours, which may be more strongly related to the exposures exam- ined here. We characterized food retailing in home and school neighbourhoods, but we did not capture exposures along the journey between home and school, or in wider activity space envi- ronments, which may be important [61–63]. We used 800m buffers around schools and home LSOA boundaries as a proxy for neighbourhoods, but this may not be an accurate reflection of a child’s neighborhood [63]. PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 11 / 17 Previous research studies on associations between food outlets around schools and obesity or diet outcomes among children have varied widely in their exposure and outcome measures [2], and have found mixed results. To the best of our knowledge, of those studies examining neighbourhood environment effects on body weight, only two have been conducted in England [73,74]. Like our study, Harrison et al considered food retail exposures in both school and home environments and found retailing around homes was more strongly associated with Fat Mass Index (FMI) than retailing around schools [63]. Among girls, access to unhealthy food outlets (takeaways and convenience stores) near home was associated with FMI and access to healthy outlets (supermarkets and greengrocers) near home was associated with a lower FMI. Similarly, we found high access to fast food in the home neighbourhoods of Year 6 students was associated with a higher BMI z-score and high access to food stores in the home of Recep- tion students was associated with a lower BMI z-score. Harrison et al also measured the child’s mode of travel to school and found associations varied between active- and non-active travel- lers, with some evidence that environmental exposures may have more of an influence on the former. This has important implications for our study because children who are driven to and from school may have little chance to interact with the school neighbourhood. Our study builds upon earlier work examining associations between food environments, deprivation and childhood overweight and obesity using NCMP data [73]. Unlike our study, which was limited to one region in England, this study included schools across England and had data from three time points (2007–2010). Our main contribution to this work is the inclu- sion of individual-level measures (rather than aggregate school-level measures). Also, while their study focused on takeaways, we considered takeaways and other types of retailers. PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 12 / 17 Food Outlets around Schools and Student BMI in England Cetateanu et al observed associations between takeaways around schools and obesity rates among older (Year 6) students, but not the younger (Reception) students. Similarly, we found stronger evidence of associations between food retailing and BMI among older students (Year 6) compared to younger students (Reception). One possible explanation is that older children have increased autonomy, spending power and capacity to travel independently to and from school. However, Sanchez et al [75] found that the presence of convenience stores near schools was significantly associated with the overweight prevalence ratio of fifth grade students (around age 10), but not ninth grade students (about 14 years). The reasons behind this dis- crepancy are currently unclear. We do not know why associations between fast food outlets near homes and mean BMI z scores were stronger for girls than they were for boys. Previous research has also shown sex dif- ferences in associations between the environment and weight-related outcomes [74,76–78], but further work is needed to understand what lies behind these differences. There are many reasons why we may not have seen an association between food retailing around schools and BMI. Perhaps an association would have been observed if we had looked at more proximal outcomes like diet or physical activity. It could be that students near schools with a high density of outlets eat more fast food, but compensate for it with higher levels of physical activity or eat less food at home. Another possibility is that the cumulative effects have not yet appeared among primary school children. Unfortunately, the NCMP doesn’t measure secondary school pupils, who may be more affected by food outlets around schools than youn- ger students. Future research on associations between food retailing and weight among second- ary schoolchildren would benefit from considering whether or not schools have stay-on-site lunch policies. While this is the norm among primary schools, many secondary schools allow older students to leave campus for meals. Future work may benefit from the collection and analysis of data over a wide range of spatial and temporal scales [13,79,80] while accounting for individual-level characteristics. Research in this area will also would benefit from using a national sample at various time points, as done by Cetateanu et al [73], while also controlling for individual-level and home-environment characteristics. Additional work is needed to find ways to more accurately characterize environmental exposures at home and school [81]. Accounting for individual mobility patterns such as activity spaces [82] or GPS routes [83] will enable researchers to assess the various environments beyond home and school. One recent example in the UK found that exposure to takeaway food outlets in home, work, and commuting environments combined was associated with higher consumption of takeaway food, greater body mass index, and greater odds of obesity among adults [55]. Obesity is the consequence of a complex web of influences; its underlying systems that are non-linear and ill-suited to dichotomous hypothesis testing [84]. The scientific evidence about the effects of food outlets on child BMI is not conclusive. However, in the face of this uncer- tainty, proponents of the precautionary principal would argue that if plausible risk [16] to health has been identified, preventive measures are warranted. S1 Table. Food Outlets in West Berkshire: Names, Postcodes, Categories and Addresses from Local Councils. (XLSX) References 1. Townsend N, Bhatnagar P, Wickramasinghe K, Williams J, Vujcich D, Rayner M (2013) Children and Young People Statistics; Weissberg P, editor. London: British Heart Foundation. 2. Williams J, Scarborough P, Matthews A, Cowburn G, Foster C, Roberts N, et al. (2014) A systematic review of the influence of the retail food environment around schools on obesity-related outcomes. 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Performed the experiments: JW PS AM NT LM TB. Analyzed the data: JW PS AM NT LM TB. Wrote the paper: JW PS J y J p p J AM NT LM TB MR. AM NT LM TB MR. PLOS ONE \| DOI:10.1371/journal.pone.0132930 July 17, 2015 References Crawford PB, Gosliner W, Kayman H (2011) Peer Reviewed: The Ethical Basis for Promoting Nutri- tional Health in Public Schools in the United States. Preventing chronic disease 8. 17. Khambalia A, Dickinson S, Hardy L, Gill T, Baur L (2012) A synthesis of existing systematic reviews and meta-analyses of school-based behavioural interventions for controlling and preventing obesity. Obesity Reviews 13: 214–233. doi: 10.1111/j.1467-789X.2011.00947.x PMID: 22070186 18. Katz D, O'connell M, Njike VY, Yeh M, Nawaz H (2008) Strategies for the prevention and control of obe- sity in the school setting: systematic review and meta-analysis. 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https://openalex.org/W2941062915	https://zenodo.org/record/3588781/files/157%20Skin%20Cancer%20Detection%20using%20Digital%20Image%20Processing%20and%20Implementation%20using%20ANN%20and%20ABCD%20Features.pdf	English	null	Skin Cancer Detection using Digital Image Processing and Implementation using ANN and ABCD Features	International journal of trend in scientific research and development	2,018	cc-by	7,566	Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma Melanoma is a serious type of skin cancer. It starts in skin cells called melanocytes. There are 3 main types of skin cancer, Melanoma, Basal and Squamous cell carcinoma. Melanoma is more likely to spread to other parts of the body. Early detection of malignant melanoma in dermoscopy images is very important and critical, since its detection in the early stage can be helpful to cure it. Computer Aided Diagnosis systems can be very helpful to facilitate the early detection of cancers for dermatologists. Image processing is a commonly used method for skin cancer detection from the appearance of affected area on the skin. In this work, a computerised method has been developed to make use of Neural Networks in the field of medical image processing. The ultimate aim of this paper is to implement cost emergency support systems; to process the medical images. It is more advantageous to patients. The dermoscopy image of suspect area of skin cancer is taken and it goes under various pre technique for noise removal and image enhancement. Then the image is undergone to segmentation using Thresholding method. Some features of image have to be extracted using ABCD rules. In this Asymmetry index and Geometric features are extracted from the segmented image. These features are given as the input to classifier. Artificial Neural Network (ANN) with feed forward architecture is used for classification purpose. It classifies the image into cancerous or non-cancerous. The proposed algorithm has been tested on the ISIC (International Skin Imaging Collaboration) 2017 training and test datasets. The ground truth data of each image is available as well, so performance of this wor evaluate quantitatively. Melanoma is a serious type of skin cancer. It starts in skin cells called melanocytes. There are 3 main types of skin cancer, Melanoma, Basal and Squamous cell is more likely to spread to other parts of the body. Early detection of malignant copy images is very important and critical, since its detection in the early stage can be helpful to cure it. Computer Aided Diagnosis elpful to facilitate the early detection of cancers for dermatologists. Image processing is a commonly used method for skin cancer detection from the appearance of affected area on the skin. In this work, a computerised method has e of Neural Networks in the field of medical image processing. Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma The ultimate aim of this paper is to implement cost-effective to process the medical It is more advantageous to patients. The of skin cancer is taken and it goes under various pre-processing technique for noise removal and image enhancement. Then the image is undergone to segmentation using sholding method. Some features of image have to be extracted using ABCD rules. In this work, Asymmetry index and Geometric features are extracted from the segmented image. These features are given as the input to classifier. Artificial Neural Network (ANN) with feed forward architecture is used for classification purpose. It classifies the given cancerous. The proposed algorithm has been tested on the ISIC (International Skin Imaging Collaboration) 2017 training and test datasets. The ground truth data of each image is available as well, so performance of this work can I. INTRODUCTION Skin cancers can be classified into melanoma and non-melanoma. Melanoma is a malignancy of the cells which gives the skin its colo and it can invade nearby tissues. Moreover, it spreads through the whole human body and it might cause to patient death and non-melanoma which is rarely spread to other parts of the human body. Malignant melanoma is the most aggressive type cancers and its incidence has been rapidly increasing [1] [2] [3] [4]. Nevertheless, it is also the most treatable type of skin cancer if detected or diagnosed at an early stage [5]. The diagnosis of melanoma in early stage is a challenging and fundamental task for dermatologists since some other skin lesions may have similar physical characteristics. Dermos considered as the widely common technique used to perform an in-vivo observation of pigmented skin lesions [6]. In early detection of malignant melanoma, dermoscopic images have great potential, but their interpretation is time consuming and subjective, even for trained dermatologists. Therefore, the need to build a system which can assist dermatologists to get right decision for their diagnosis has become very important. Skin cancers can be classified into melanoma and melanoma. Melanoma is a malignancy of the cells which gives the skin its colour (melanocytes) and it can invade nearby tissues. Moreover, it spreads through the whole human body and it might cause to melanoma which is rarely spread to other parts of the human body. Malignant melanoma is the most aggressive type of human skin cancers and its incidence has been rapidly increasing [1] [2] [3] [4]. International Journal of Trend in International Open Access Journal ISSN No: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) International Open Access Journal \| www.ijtsrd.com ISSN No: 2456 - 6470 \| Volume - 2 \| Issue – 6 \| Sep Research and Development (IJTSRD) www.ijtsrd.com 6 \| Sep – Oct 2018 International Journal of Trend in International Open Access Journal ISSN No: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) International Open Access Journal \| www.ijtsrd.com ISSN No: 2456 - 6470 \| Volume - 2 \| Issue – 6 \| Sep Research and Development (IJTSRD) www.ijtsrd.com 6 \| Sep – Oct 2018 ABSTRACT Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma II. Methodology The following steps are implemented for classification of skin cancer. The following steps are implemented for classification The following steps are implemented for classification of skin cancer. The following steps are implemented for classification Image Acquisition Pre-processing S t ti Pre-processing Segmentation Feature Extraction Classification Feature Extraction Classification Th fi i h Th fi i h The computer is not more inventive than human but probably it may be able to extract some information, like colour variation, asymmetry, texture features, more accurately that may not be readily observed by naked human eyes [5]. There have been many proposed systems and algorithms such as the s point checklist, ABCD rule, and the Menzies [2, 3] to improve the diagnostics of the melanoma skin cancer. The computer is not more inventive than human but le to extract some information, like colour variation, asymmetry, texture features, more accurately that may not be readily observed by 5]. There have been many proposed systems and algorithms such as the seven- and the Menzies method ] to improve the diagnostics of the melanoma The first step is the capture image is acquired from the ISI through MATLAB. The second step is image pre processing, the pre-processing technique can be applied to eliminate the irrelevant data contains in the image. The skin lesion regions such as oils, hairs are removed from the original image using median filter techniques. The third step is image segmentation: the goal of image segmentation is to make simpler change the representation of an image into something that is more meaning analyze. The next step is image enhancement; to improve the quality of the image so that the consequential image is better than the original image. In this step, the required region is segmented out and detected the edge of the skin lesio feature extraction and the final step is classification of the skin lesion image. The first step is the capture image the skin lesion image is acquired from the ISIC 2018 database through MATLAB. The second step is image pre- processing technique can be applied to eliminate the irrelevant data contains in the image. The skin lesion regions such as label, marks, oils, hairs are removed from the original image using median filter techniques. International Journal of Trend in Scientific Research and Development (IJTS International Journal of Trend in Scientific Research and Development (IJTS International Journal of Trend in Scientific Research and Development (IJTS International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 accuracy depends on the factor of training the dermatologist. accuracy depends on the factor of training the computerised melanoma detection using Artificial Neural Network classification has been adapted which is efficient than the conventional one and Melanoma detection using Artificial Neural Network is a more effective method compared to other. computerised melanoma detection using Artificial Neural Network classification has been adapted which is efficient than the conventional one and Melanoma Neural Network is a more effective method compared to other. computerised melanoma detection using Artificial Neural Network classification has been adapted which is efficient than the conventional one and Melanoma detection using Artificial Neural Network is a more effective method compared to other. computerised melanoma detection using Artificial Neural Network classification has been adapted which is efficient than the conventional one and Melanoma Neural Network is a more effective method compared to other. The diagnosis of melanoma from melanocytic nevi is not clear and easy to identify, especially in the early stage. Thus, automatic diagnosis tool is more effective and essential part of physicians. Even when the dermoscopy for diagnosis is done with the expert dermatologists, the accuracy of melanoma diagnosis is not more than 75-84% [4]. The computer aided diagnostics is more useful to increase the diagnosis accuracy as well as the speed [5]. iagnosis of melanoma from melanocytic nevi is not clear and easy to identify, especially in the early stage. Thus, automatic diagnosis tool is more effective and essential part of physicians. Even when the dermoscopy for diagnosis is done with the expert ermatologists, the accuracy of melanoma diagnosis 4]. The computer aided diagnostics is more useful to increase the diagnosis II. Methodology The third step is image segmentation: the goal of image segmentation is to make simpler change the representation of an image into something that is more meaningful and easier to analyze. The next step is image enhancement; to improve the quality of the image so that the consequential image is better than the original image. In this step, the required region is segmented out and detected the edge of the skin lesion. The fourth step is feature extraction and the final step is classification of The key steps in a computer-aided diagnosis of melanoma skin cancer are image acquisition of a skin lesion, segmentation of the skin lesion from skin region, extraction of geometric features of the lesion blob and feature classification. Segmentation or border detection is the course of action of separating the skin lesion of melanoma from the circumferential skin to form the area of interest. Feature extraction is done to extract the geometric features which are accountable for increasing the accuracy; corresponding to those visually detected by dermatologists, that meticulously characterizes a melanoma lesion. The feature extraction methodology of many aided diagnosis of melanoma skin cancer are image acquisition of a skin lesion, segmentation of the skin lesion from skin geometric features of the lesion blob and feature classification. Segmentation or border detection is the course of action of separating the skin lesion of melanoma from the circumferential skin to form the area of interest. Feature extraction is xtract the geometric features which are accountable for increasing the accuracy; ally detected by that meticulously characterizes a The feature extraction methodology of many Fig. 1System Block Diagram System Block Diagram image The feature extraction methodology of many computerised melanoma detection systems has been largely depending on the conventional clinical diagnostic algorithm of ABCD-rule of dermoscopy due to its effectiveness and simplicity of implementation [7]. The effectiveness of methodology stems from the fact that it incorporates the classic features of a melanoma lesion such as asymmetry, border irregularity, colour and diameter (or differential structures), where surveyable measures can be computed. The feature extraction methodology of many detection systems has been largely depending on the conventional clinical rule of dermoscopy simplicity of ]. The effectiveness of methodology tes the classic features of a melanoma lesion such as asymmetry, border irregularity, colour and diameter (or differential structures), where surveyable measures Fig. 1System Block Diagram System Block Diagram image International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 Keyword: Skin cancer, Segmentation, Feature Extraction, Classification, Melanoma n cancer, Segmentation, Feature Classification, Melanoma Nevertheless, it is also the most treatable type of skin cancer if detected or diagnosed at an early stage [5]. The diagnosis of melanoma in and fundamental task for dermatologists since some other skin lesions may have similar physical characteristics. Dermoscopy is considered as the widely common technique used to vivo observation of pigmented skin n of malignant melanoma, dermoscopic images have great potential, but their interpretation is time consuming and subjective, even for trained dermatologists. Therefore, the need to build a system which can assist dermatologists to get eir diagnosis has become very Image processing is one of the widely used methods for skin cancer detection. Dermoscopy could be a non-invasive examination technique supported the cause of incident light beam and oil immersion technique to form potential the visual investigation of surface structures of the skin. The detection of melanoma using dermoscopy is higher than individual observation based detection [ Image processing is one of the widely used methods for skin cancer detection. Dermoscopy could be a invasive examination technique supported the cause of incident light beam and oil immersion otential the visual investigation of surface structures of the skin. The detection of melanoma using dermoscopy is higher than individual detection [3], but its diagnostic @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 Oct 2018 Page: 962 B. Skin Lesion Segmentation 1. Area (A): Number of pixels of the lesion. A): Number of pixels of the lesion. g Image segmentation is an essential process for most image analysis subsequent tasks. Segmentation divides an image into its constituent regions or objects. The goal of segmentation is to make simpler or change the representation of an image into something that is more meaningful and easier to analyse. Image segmentation is an essential process for most image analysis subsequent tasks. Segmentation image into its constituent regions or objects. The goal of segmentation is to make simpler or change the representation of an image into something that is more meaningful and easier to 2. Perimeter (P): Number of pixels detected boundary Number of pixels along the Image segmentation is an essential process for most image analysis subsequent tasks. Segmentation divides an image into its constituent regions or objects. The goal of segmentation is to make simpler or change the representation of an image into something that is more meaningful and easier to analyse. Image segmentation is an essential process for most image analysis subsequent tasks. Segmentation image into its constituent regions or objects. The goal of segmentation is to make simpler or change the representation of an image into something that is more meaningful and easier to 3. Greatest Diameter (GD): The length of the line which connects the two farthest ): The length of the line connects the two farthest 4. Shortest Diameter (SD): The length of the line connecting the two closest boundary points and passes across the lesion centroid. Shortest Diameter (SD): The length of the line connecting the two closest boundary points and passes across the lesion centroid. 5. Circularity Index (CRC): It explains the shape uniformity. Circularity Index (CRC): It explains the shape Image segmentation is the course of action of segregating an image into multiple parts, which is used to identify objects or other relevant information in digital images. Background subtraction, also known as blob detection, is an emerging technique in the fields of image processing wherein an image’s foreground is extracted for further processing. Typically, an image’s regions of interest are objects in its foreground. Segmentation methods can be classified as thresholding, region based, Edge based and clustering. In this work, thresholding is used because of the computationally inexpensive and fast and simple to implement. III. Performance Evaluation Parameter Performance Evaluation Parameter The en positive class and negative class predictions can be depicted as a 2 2 confusion matrix that tabulates whether predictions fall into one To prevent edges which are dark in many of the images, from remaining as part of the lesion mask; dark in many of the images, from remaining as part of the lesion mask; To prevent edges which are dark in many of the images, from remaining as part of the lesion mask; To reduce any effects of disturbing artifacts as far as possible; dark in many of the images, from remaining as part of the lesion mask; To reduce any effects of disturbing artifacts as far To prevent edges which are dark in many of the images, from remaining as part of the lesion mask; dark in many of the images, from remaining as part of the lesion mask; To prevent edges which are dark in many of the images, from remaining as part of the lesion mask; To reduce any effects of disturbing artifacts as far as possible; dark in many of the images, from remaining as part of the lesion mask; To reduce any effects of disturbing artifacts as far g g p To reduce any effects of disturbing artifacts as far as possible; g g p To reduce any effects of disturbing artifacts as far g g p To reduce any effects of disturbing artifacts as far as possible; g g p To reduce any effects of disturbing artifacts as far To set soft edges, to ensure there are not too many recesses or projections and that there is certain convexity in the resulting mask To set soft edges, to ensure there are not too many recesses or projections and that there is certain To set soft edges, to ensure there are not too many recesses or projections and that there is certain convexity in the resulting mask To set soft edges, to ensure there are not too many recesses or projections and that there is certain True positives (TP): These refer to the positive tuples that were correctly label classifier. It is assumed that TP is the number of true positives. P): These refer to the positive tuples that were correctly labelled by the assifier. III. Performance Evaluation Parameter Performance Evaluation Parameter the image then needs to be post processed in order to resolve any problems there may be as follows: the image then needs to be post processed in order to resolve any problems there may be as follows: The most common performance measures consider the model’s ability to discern one class versus all others. The class of interest is known as the positive class, while all others are known as negative. The relationship between positive class and negative class predictions can be depicted as a 2 matrix that tabulates whether predictions fall into one of four categories: The most common performance measures consider the model’s ability to discern one class versus all others. The class of interest is known as the positive class, while all others are known as negative. III. Performance Evaluation Parameter Performance Evaluation Parameter It is assumed that TP is the number of These problems can be reduces by using morphological operation such as opening, closing and filling process. These problems can be reduces by using morphological operation such as opening, closing and True negatives (TN): These are the negative tuples that were correctly labelled by the classifier. It is assumed that TN is the number of true negatives. True negatives (TN): These are the negative tuples ed by the classifier. It is assumed that TN is the number of true negatives. E. Classification The main issue of the classification task is to avoiding over fitting caused by the sma skin lesion in most dermatology datasets. In order to solve this problem, the objective of the proposed model is to firstly extract features from images and secondly load those extracted representations on ANN network to classify. The main issue of the classification task is to avoiding over fitting caused by the small number of images of skin lesion in most dermatology datasets. In order to solve this problem, the objective of the proposed model is to firstly extract features from images and extracted representations on an C. Post-processing Once binary image of skin lesion has been obtained, the image then needs to be post processed in order to resolve any problems there may be as follows: Once binary image of skin lesion has been obtained, the image then needs to be post processed in order to resolve any problems there may be as follows: III. Performance Evaluation Parameter Performance Evaluation Parameter B. Skin Lesion Segmentation Image segmentation is the course of action of image into multiple parts, which is used to identify objects or other relevant information in digital images. Background subtraction, also known as blob detection, is an emerging technique in the fields of image processing wherein an image’s extracted for further processing. Typically, an image’s regions of interest are objects in its foreground. Segmentation methods can be classified as thresholding, region based, Edge based thresholding is used omputationally inexpensive and fast Image segmentation is the course of action of segregating an image into multiple parts, which is used to identify objects or other relevant information in digital images. Background subtraction, also known as blob detection, is an emerging technique in the fields of image processing wherein an image’s foreground is extracted for further processing. Typically, an image’s regions of interest are objects in its foreground. Segmentation methods can be classified as thresholding, region based, Edge based and clustering. In this work, thresholding is used because of the computationally inexpensive and fast and simple to implement. Image segmentation is the course of action of image into multiple parts, which is used to identify objects or other relevant information in digital images. Background subtraction, also known as blob detection, is an emerging technique in the fields of image processing wherein an image’s extracted for further processing. Typically, an image’s regions of interest are objects in its foreground. Segmentation methods can be classified as thresholding, region based, Edge based thresholding is used omputationally inexpensive and fast 6. Irregularity Index A (IrA): Irregularity Index A (IrA): 7. Irregularity Index B (IrB): 8. Irregularity Index C (IrC): 9. Irregularity Index D (IrD): Irregularity Index D (IrD): Diameter: Diameter in pixels Diameter in pixels end in Scientific Research and Development (IJTSRD) ISSN: 2456 end in Scientific Research and Development (IJTSRD) ISSN: 2456 end in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 A. Pre-processing p g In this work, few previous processing techniques are needed. Prior to segmentation, all processed in order to minimize undesirable features that could affect the performance of the algorithm such as reflections, presence of the hair and colo differences between images. The image is also normalized to a unique size and shape. normalization allows for the comparison of the region features such as positions and sizes between different images. In this work, few previous processing techniques are needed. Prior to segmentation, all images are pre- processed in order to minimize undesirable features that could affect the performance of the algorithm such as reflections, presence of the hair and colour differences between images. The image is also normalized to a unique size and shape. This normalization allows for the comparison of the region features such as positions and sizes between different Dermoscopy is a diagnostic technique that is used worldwide in the recognition and interpretation of copious skin lesions [4]. Other than dermoscopy, a Dermoscopy is a diagnostic technique that is used n and interpretation of copious skin lesions [4]. Other than dermoscopy, a @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 Oct 2018 Page: 963 D. Feature Extraction The foremost features of the Melanoma Skin Lesion are its Asymmetric Index, Border features, Colo Diameter. Hence, this system is proposed to extract the (9) Geometric Features, (1) Asymmetry Index and (1) diameter of the segmented skin lesion. These features are adopted from the segmented image containing only skin lesion, the image blob of the skin lesion is analyzed to extract the 11 features The foremost features of the Melanoma Skin Lesion are its Asymmetric Index, Border features, Colour and Diameter. Hence, this system is proposed to extract the (9) Geometric Features, (1) Asymmetry Index and (1) diameter of the segmented skin lesion. These features are adopted from the segmented image containing only skin lesion, the image blob of the skin act the 11 features. False positive (FP): These are the ne that were incorrectly labe assumed that FP is the number of false positives. False positive (FP): These are the negative tuples that were incorrectly labelled as positive. It is assumed that FP is the number of false positives. False negative (FN): These are the positive tuples that were mislabelled as negative. It is assumed that FN is the number of false negatives. False negative (FN): These are the positive tuples led as negative. It is assumed that FN is the number of false negatives. False negative (FN): These are the positive tuples that were mislabelled as negative. It is assumed that FN is the number of false negatives. False negative (FN): These are the positive tuples led as negative. It is assumed that FN is the number of false negatives. IV. Test And Results A. Results of Segmentation Process There are three main phases: namely pre segmentation and post-processing in segmentation part. The input of the system is dermoscopic image o skin lesion as shown in Fig. 2. After resizing the input image, this image will convert to the gray scale in order to get grater separability between the lesion and background healthy skin. The resultant image has been displayed. Most of the dermoscopic images have some artifacts such as oil, bubble hair, noise etc. these artifacts are removed by median filter on gray scale image. After this, the Otsu’s thresholding method has been applied on the gray scale intensity image. This gives the desired segmented image. namely pre-processing, processing in segmentation part. The input of the system is dermoscopic image of . After resizing the input gray scale image in order to get grater separability between the lesion and background healthy skin. The resultant gray scale image has been displayed. Most of the dermoscopic images have some artifacts such as oil, bubble hair, ved by applying scale image. After this, the Otsu’s thresholding method has been applied on the scale intensity image. This gives the desired The first stage of the system is to select skin lesion images. This can click original image menu item from main window GUI of the proposed system. The first stage of the system is to select skin lesion This can click original image menu item from main window GUI of the proposed system. After loading the input image, it is needed to segment the lesion image by using O Technique. The segmented image obtained from Otsu's thresholding has the advantages of smaller storage space, fast processing speed and ease in manipulation, compared with gray usually contains 256 levels. After loading the input image, it is needed to segment the lesion image by using Otsu's Thresholding Technique. The segmented image obtained from Otsu's thresholding has the advantages of smaller storage space, fast processing speed and ease in n, compared with gray level image which In feature extraction, the standard features such as Asymmetry Index, Area, Perimeter, Major Axis Length, Minor Axis length, Circularity Index, Irregularity Index and diameter are segmented test image as shown in Fig. standard features are very useful to classify the melanoma skin cancer more accurately. Accuracy can be calculated by using this e racy can be calculated by using this equation: Accuracy can be calculated by using this e racy can be calculated by using this equation: Asymmetry Features: Major and Minor Asymmetry Indices: Asymmetry Features: Major and Minor Asymmetry Accuracy (1) (1) Geometric Features: Geometric Features: @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 Oct 2018 Page: 964 @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 TP TN TP TN FP FN International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 d in Scientific Research and Development (IJTSRD) ISSN: 2456 FN racy only measures of the classifier and dictions. is computed as the rrectly identified. d in Scientific Research and Development (IJTSRD) ISSN: 2456 racy only measures of the classifier and dictions. is computed as the rrectly identified. (2) Fig.2. Testing result of pre segmentation d in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 result of pre-processing and segmentation nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 nd Development (IJTSRD) ISSN: 2456 nd Development (IJTSRD) ISSN: 2456 Fig.2. Testing result of pre segmentation nd Development (IJTSRD) ISSN: 2456-6470 result of pre-processing and segmentation As indicated in Equation (1), accuracy only measures the number of correct predictions of the classifier and ignores the number of incorrect predictions. , accuracy only measures the number of correct predictions of the classifier and ignores the number of incorrect predictions. Sensitivity, also known as recall, is computed as the fraction of true positives that are correctly identified. , is computed as the s that are correctly identified. Fig.2. Testing result of pre segmentation result of pre-processing and segmentation FN TP TP y Sensitivit + = (2) (2) Precision, which is computed as the fraction of retrieved instances that are relevant. which is computed as the fraction of Precision, which is computed as the fraction of retrieved instances that are relevant. which is computed as the fraction of FP TN TN y Specificit + = IV. Test And Results extraction, the standard features such as Asymmetry Index, Area, Perimeter, Major Axis Length, Minor Axis length, Circularity Index, Irregularity Index and diameter are extracted from the ted test image as shown in Fig.6. These ery useful to classify the melanoma skin cancer more accurately. After these steps morphological operations are used to enhance the segmented image. In this research work, morphological opening, dilation, erosion and closing operations are used. A morphological closing of the mask is performed using a disk of radius 500 pixels followed by a dilation using a disk of radius 40 pixels. These operations smooth the border of the mask. The experimental results for this post-processing phase are shown in Fig. 3. After these steps morphological operations are used to ented image. In this research work, morphological opening, dilation, erosion and closing operations are used. A morphological closing of the med using a disk of radius 500 pixels followed by a dilation using a disk of radius 40 pixels. operations smooth the border of the mask. The essing phase are After extracting the features, these features are given as input to the classifier. The classifier produces whether the image is Melanoma or not. For the Melanoma condition, the classifier o normal skin (not melanoma) the output is 0. By pressing the classify button, the classification process is performed and the result is displayed as shown in Fig. 7 and 8. After extracting the features, these features are given er. The classifier produces whether the image is Melanoma or not. For the Melanoma condition, the classifier output is 1 and for normal skin (not melanoma) the output is 0. By pressing the classify button, the classification process ult is displayed as shown in @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 Oct 2018 Page: 965 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 Fig.4. Loading Input Image Fig.5. After Segmentation International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 4. Loading Input Image 5. After Segmentation Fig.7. Classification Results of Melanoma Fig.8. Classification Results of Non C. Performance Evaluation To evaluate performance in this system, 40 images from a test data set are tested. The accuracy of skin lesion classification system is calculated. Table 1 describes the performance of the proposed system. IV. Test And Results TABLE I Performance of The Proposed System N o Im age Set TP F P T N F N Acc urac y International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 7. Classification Results of Melanoma 8. Classification Results of Non-Melanoma performance in this system, 40 unknown images from a test data set are tested. The accuracy of system is calculated. Table describes the performance of the proposed system. Performance of The Proposed System Acc urac y Sensi tivity Pre cisi on Spe cific ity nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 nal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 @ IJTSRD \| Available Online @ www.ijtsrd.com Fig.4. Loading Input Image Fig.5. After Segmentation Fig.6. Extracted Features from Testing Image www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 4. Loading Input Image 5. After Segmentation Fig.6. Extracted Features from Testing Image Fig.7. Classification Results of Melanoma Fig.8. Classification Results of Non C. Performance Evaluation To evaluate performance in this system, 40 images from a test data set are tested. The accuracy of skin lesion classification system is calculated. Table 1 describes the performance of the proposed system. TABLE I Performance of The Proposed System N o . Im age Set TP F P T N F N Acc urac y 1 Tes t ing Set 18 2 19 1 92.5 % Oct 2018 Page: 966 7. Classification Results of Melanoma 8. Classification Results of Non-Melanoma performance in this system, 40 unknown images from a test data set are tested. The accuracy of system is calculated. Table describes the performance of the proposed system. Performance of The Proposed System Acc urac y Sensi tivity Pre cisi on Spe cific ity 92.5 % 94.7 % 90 % 90.4 7% Fig.4. Loading Input Image 4. Loading Input Image Fig.4. Loading Input Image 4. Loading Input Image Fig.7. Classification Results of Melanoma 7. Classification Results of Melanoma Fig.7. Classification Results of Melanoma 7. Classification Results of Melanoma Fig.4. Loading Input Image Fig.5. After Segmentation 4. Loading Input Image 5. After Segmentation Fig.7. References All the features have been calculated based on Otsu’s segmentation method. Using the Neural Network classifier, melanoma skin cancer diagnosis with a training accuracy of 100% and testing accuracy of 93% is achieved. Computational time is around 14 seconds fo lesion classification. For illustration, a graphical user interface has developed in order to facilitate the diagnostic task for the dermatologists. In this work, the system for melanoma skin cancer detection system is developed by using MATLAB. step in early detection of melanoma skin cancer. To analyze skin lesions, it is necessary to accurately locate and isolate the lesions. In this thesis work, the Otsu’s method is the oldest and simplest one. It has shown the best the three methods. Feature extraction is considered as the most critical art skin cancer screening system. In this d on ABCD-rule Algorithms for extracting features the features have been calculated based on Otsu’s segmentation method. Using the Neural Network classifier, melanoma skin cancer diagnosis with a training accuracy of 100% and testing accuracy of 93% is achieved. Computational time is around 14 seconds for each lesion classification. For illustration, a graphical user interface has developed in order to facilitate the 1. Xu L, Jackowski M, Goshtasby A, Roseman, “Segmentation of cancer images”, Image and Vision Computing, Volume 17, Issue1, pg. 65 ckowski M, Goshtasby A, Roseman, tation of cancer images”, Image and Vision Computing, Volume 17, Issue1, pg. 65-74. 2. J Abdul Jaleel, Sibi Salim, Aswin. “Computer Aided Detection 01 Skin Cancer”, International Conference on Circuits, Power and Computing Technologies, 2013 J Abdul Jaleel, Sibi Salim, Aswin. R. B, “Computer Aided Detection 01 Skin Cancer”, International Conference on Circuits, Power and Computing Technologies, 2013 3. Arroyo J L G and Zapirain B G Detection 01 Skin Cancer”, Computers in biology and biology and medicine 44 14157 and Zapirain B G, “Computer Aided Detection 01 Skin Cancer”, 2014, Computers in Computers in biology and .medicine 44 14157 biology and medicine 44 14157 4. Dr. S. Gopinathan, S. “Feature Extraction through Image Processing Techniques”, International Journal of Emerging Trends & Technology in Computer Science (IJETTCS), Volume 5, Issue 4, July Web Site:www.ijettcs.org Nancy Arokia Rani, “Feature Extraction through Image Processing national Journal of Emerging Trends & Technology in Computer Science sue 4, July-August 2016, Web Site:www.ijettcs.org 5. IV. Test And Results Classification Results of Melanoma Fig.8. Classification Results of Non 7. Classification Results of Melanoma 8. Classification Results of Non-Melanoma Fig.5. After Segmentation 5. After Segmentation Fig.5. After Segmentation 5. After Segmentation Fig.5. After Segmentation 5. After Segmentation Fig.8. Classification R Performance Evalua 8. Classification R Fig.6. Extracted Features from Testing Image Fig.6. Extracted Features from Testing Image To evaluate performance in this system, 40 images from a test data set are tested. The accuracy of skin lesion classification system is calculated. Table 1 describes the performance of the proposed system. performance in this system, 40 unknown images from a test data set are tested. The accuracy of system is calculated. Table describes the performance of the proposed system. TABLE I Performance of The Proposed System N o . Im age Set TP F P T N F N Acc urac y 1 Tes t ing Set 18 2 19 1 92.5 % Performance of The Proposed System Acc urac y Sensi tivity Pre cisi on Spe cific ity 92.5 % 94.7 % 90 % 90.4 7% Fig.6. Extracted Features from Testing Image Fig.6. Extracted Features from Testing Image Fig.6. Extracted Features from Testing Image Fig.6. Extracted Features from Testing Image International Journal of Trend in Scientific Research and Development (I International Journal of Trend in Scientific Research and Development (I International Journal of Trend in Scientific Research and Development (I International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 References Uzma Bano Ansari, “Skin Cancer Detection Using Image Processing”, International Research Journal of Engineering and Technology (IRJET), Volume: 04 \| Apr-2017 Uzma Bano Ansari, “Skin Cancer Detection Using Image Processing”, International Research Journal and Technology (IRJET), Volume: References In this work, the system for melanoma skin cancer detection system is developed by using MATLAB. Image segmentation is the first step in early detection of melanoma skin cancer. To analyze skin lesions, it is necessary to accurately locate and isolate the lesions. In this thesis work, the Otsu’s method is the oldest and simplest one. It has shown the best segmentation results among the three methods. Feature extraction is considered as the most critical state–of-the- art skin cancer screening system. In this thesis, the feature extraction is based on ABCD of dermatoscopy. Algorithms for extracting features have been disused. All the features have been calculated based on Otsu’s segmentation method. Using the Neural Network classifier, melanoma skin cancer diagnosis with a training accuracy of 100% and testing accuracy of 93% is achieved. Computational time is around 14 seconds fo lesion classification. For illustration, a graphical user interface has developed in order to facilitate the diagnostic task for the dermatologists. In this work, the system for melanoma skin cancer detection system is developed by using MATLAB. step in early detection of melanoma skin cancer. To analyze skin lesions, it is necessary to accurately locate and isolate the lesions. In this thesis work, the Otsu’s method is the oldest and simplest one. It has shown the best the three methods. Feature extraction is considered as the most critical art skin cancer screening system. In this d on ABCD-rule Algorithms for extracting features the features have been calculated based on Otsu’s segmentation method. Using the Neural Network classifier, melanoma skin cancer diagnosis with a training accuracy of 100% and testing accuracy of 93% is achieved. Computational time is around 14 seconds for each lesion classification. For illustration, a graphical user interface has developed in order to facilitate the In this work, the system for melanoma skin cancer detection system is developed by using MATLAB. Image segmentation is the first step in early detection of melanoma skin cancer. To analyze skin lesions, it is necessary to accurately locate and isolate the lesions. In this thesis work, the Otsu’s method is the oldest and simplest one. It has shown the best segmentation results among the three methods. Feature extraction is considered as the most critical state–of-the- art skin cancer screening system. In this thesis, the feature extraction is based on ABCD of dermatoscopy. Algorithms for extracting features have been disused. Acknowledgment Firstly, the author would like to acknowledge particular thanks to Union Minister of the Ministry of Science and Education, for permitting to attend the Master program at Pyay Technology University. Much gratitude is owed to Dr. Nyaunt Soe, Rector, Pyay Technological University, for his kind permission to carry out this paper. The author i deeply thankful to her supervisor, Dr. Moe Mon Myint, Professor, Department of Electronic Engineering, Pyay Technological University, fo helpful and for providing guidelines. Moreover, the author wishes to express special thanks to Dr. Khin Thu Zar Win, Professor and Head, Department of Mechatronic Engineering, Pyay Technological University, Pyay for her kindness and suggestions. Finally, I would like to thank my parents for supporting to me. Firstly, the author would like to acknowledge the Ministry of Science and Education, for permitting to attend the Master program at Pyay Technology University. Much gratitude is owed to Dr. Nyaunt Soe, Rector, Pyay Technological University, for his kind permission to carry out this paper. The author is r supervisor, Dr. Moe Mon Professor, Department of Electronic Engineering, Pyay Technological University, for her guidelines. Moreover, the author wishes to express special thanks to Dr. Khin in, Professor and Head, Department of Mechatronic Engineering, Pyay Technological University, Pyay for her kindness and suggestions. Finally, I would like to thank my parents for 6. Shivangi jain, Vandana jagtap, Nitin Pise, “Computer Aided Melanoma Skin Cancer Detection Using Image Processing”, Internal Conference on Intelligent Computing Communication & Convergence, Vol 48, pp 725 740, 2015, ISSN 1877 http://www.sciencedirect.com/science/article/pii/S 1877050915007188 Shivangi jain, Vandana jagtap, Nitin Pise, “Computer Aided Melanoma Skin Cancer Detection Using Image Processing”, Internal Conference on Intelligent Computing Communication & Convergence, Vol 48, pp 725- 015, ISSN 1877-0509, .sciencedirect.com/science/article/pii/S 6. Shivangi jain, Vandana jagtap, Nitin Pise, “Computer Aided Melanoma Skin Cancer Detection Using Image Processing”, Internal Conference on Intelligent Computing Communication & Convergence, Vol 48, pp 725 740, 2015, ISSN 1877 http://www.sciencedirect.com/science/article/pii/S 1877050915007188 Shivangi jain, Vandana jagtap, Nitin Pise, “Computer Aided Melanoma Skin Cancer Detection Using Image Processing”, Internal Conference on Intelligent Computing Communication & Convergence, Vol 48, pp 725- 015, ISSN 1877-0509, .sciencedirect.com/science/article/pii/S IS irect.co Comput IS irect.co Comput 7. M. Elbaum, “Computer diagnosis”, Dermatologic clinics, vol. 20, pp. 735 747, 2002 M. Elbaum, “Computer-aided melanoma diagnosis”, Dermatologic clinics, vol. 20, pp. 735- 7. M. Elbaum, “Computer diagnosis”, Dermatologic clinics, vol. 20, pp. 735 747, 2002 M. Acknowledgment Elbaum, “Computer-aided melanoma diagnosis”, Dermatologic clinics, vol. 20, pp. 735- @ IJTSRD \| Available Online @ www.ijtsrd.com www.ijtsrd.com \| Volume – 2 \| Issue – 6 \| Sep-Oct 2018 Oct 2018 Page: 967
https://openalex.org/W2778343365	https://iris.uniroma1.it/bitstream/11573/1663416/1/Harrath_Food-restriction-during-pregnancy_2019.pdf	English	null	Food restriction during pregnancy and female offspring fertility: adverse effects of reprogrammed reproductive lifespan	Journal of ovarian research	2,017	cc-by	7,500	Abstract Background: Food restriction during pregnancy can influence the health of the offspring during the adulthood. The aim of the present study was to examine the effect of maternal food restriction (MFR) on the reproductive performance in female rat offspring from the first (FR1) and second (FR2) generations. Methods: Adult virgin Wistar female rats were given free access to tap water and were fed ad libitum on standard rodent chow, were mated with virgin adult males, and then were randomly divided into two groups: controls (that was fed ad libitum) and food-restricted group (FR, that was given only 50% of ad libitum food throughout gestation). Their first (FR1) and the second (FR2) generation of offspring were fed ad libitum and sacrificed before puberty and at adulthood. Their ovaries were removed and their histology evaluated by estimating the number of follicles (total and at various stages of folliculogenesis), and the presence of multi-nuclei oocytes and multi-oocyte follicles. Results: Total number of ovarian follicles was lower in FR1 females at week 4 in comparison with controls, while it was not different in FR2 females vs. controls. The number of the primordial follicle was lower in FR1 and FR2 females vs. controls at both week 4 and at week 8. When compared to the controls, the follicles containing multi-nuclei oocytes were more frequent in ovaries from FR1 and FR2 females at week 4, and higher and lower respectively in ovaries form FR1 and FR2 females at week 8. Conclusion: MFR affects ovarian histology by inducing the development of abnormal follicles in the ovaries in first and second generation offspring. This finding could influence the ovarian function resulting in an early pubertal onset and an early decline in reproductive lifespan. Keywords: Follicle, Food restriction, Ovary, Oocyte, Pregnancy Keywords: Follicle, Food restriction, Ovary, Oocyte, Pregnancy of common diseases may be due to the environment that the fetus directly senses via the mother [2, 5–8]. In par- ticular, maternal food restriction (MFR) has been associ- ated with coronary heart disease and increased arterial blood pressure [2], reduced nephron endowment and in- creased renal morbidity in adulthood [9], and may affect physical growth and neurobehavior in newborns [10]. Malnutrition during gestation has been associated with hepatic steatosis, type 2 diabetes, and obesity during adult- hood [5, 11–15]. * Correspondence: hharrath@ksu.edu.sa 1Zoology Department, College of Science, King Saud University, Riyadh, Saudi Arabia 2Unit of Reproductive and Developmental Biology, Faculty of Science of Tunis University of Tunis El Manar, Tunis, Tunisia Full list of author information is available at the end of the article © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Abstract During late gestation, MFR is associated with metabolic signaling dysfunction in the liver, and pre- disposes the offspring to insulin resistance [5]. Food restriction during pregnancy and female offspring fertility: adverse effects of reprogrammed reproductive lifespan Abdel Halim Harrath1,2* , Abdulkarem Alrezaki1, Lamjed Mansour1, Saleh H. Alwasel1 and Stefano Palomba3 Harrath et al. Journal of Ovarian Research (2017) 10:77 DOI 10.1186/s13048-017-0372-x Harrath et al. Journal of Ovarian Research (2017) 10:77 DOI 10.1186/s13048-017-0372-x Background Maternal nutritional status during gestation is a key de- terminant for the health and physiology of the offspring at adulthood, and that influence is mainly established during early development, i.e. well before birth [1–5]. Many adulthood diseases can be linked to the environ- ment within which the embryo has developed, including abnormal nutritional, environmental, and hormonal in- sults that may have changed the developmental trajectory of the fetus [1, 2]. According to this hypothesis, the origins Oocyte quality is a critical determinant for the devel- opmental trajectory of the fetus [16, 17]. Many forms of female reproductive disruptions have been linked to the Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 2 of 9 prenatal environment, and it is likely related to early oo- cyte formation, which is vulnerable to numerous envir- onmental effects [4]. In comparison with girls who were born appropriate for the gestational age, girls born small for the gestational age have a reduced reproductive life- span, indicated by a decrease in ovarian size that is asso- ciated with low ovulation rates [18, 19], advanced menarche, and early menopause [20–22]. Polycystic ovarian syndrome (PCOS), one of the most common fe- male endocrine disorders [23], has been suggested to arise through a gene–environment interaction, probably in the developmental milieu within which female gam- etes are formed [24]. Although maternal malnutrition is a major factor that adversely affects fetal growth and is associated with to lifelong consequences, relatively few studies have investigated the effects of MFR on the re- productive outcomes of offspring [4, 25, 26]. In a large epidemiological study women born to mothers exposed to famine were more reproductively successful compared to controls [27]. Moreover, malnutrition during preg- nancy alters reproduction in sheep by inducing poor oo- cyte quality, may cause reproductive disruptions in rats mainly by an early vaginal opening, and induce a decrease in the primordial and antral follicle number [28–30]. During the first trimester of pregnancy in cattle, MFR leads to a reduced ovarian reserve in adulthood, as observed by the increased follicle stimulating hormone (FSH) levels [31]. generation of offspring (FR1) were fed ad libitum. After complete weaning, FR1 and control females were sacri- ficed before puberty (week 4, n. 10) and at adulthood (week 8, n. 10). The ovaries were removed and the fat was discarded. Light microscopy O f Ovaries were fixed in neutral buffer formalin (NBF) or Bouin’s fluid, and were subsequently preserved in 70% alcohol. At least three ovaries from each group have been cut in serial sections to a thickness of 7 μm using a Reichert-Jung microtome. Hematoxylin and eosin (H&E) staining was used to assess ovarian architecture in the group C, FR1, and FR2. The effects of nutrition on folliculogenesis were evalu- ated by counting the number of primordial, primary, secondary, pre-antral, and antral follicles with visible oo- cyte nuclei in some slides for each ovary (see below). Specifically, ovarian follicles were classified according to the scheme of Pedersen and Peters (1968), with modifi- cations. Primordial follicles included oocytes surrounded by a single layer of three to six squamous epithelial cells, whereas primary follicles were composed of oocytes sur- rounded by one layer of numerous cuboidal epithelial cells. Secondary follicles were characterized by oocytes surrounded by more than one layer of granulosa cells with no visible antral spaces. Antral follicles were com- posed of an oocyte surrounded by many layers of cu- boidal granulosa cells, with many visible small antral spaces, or one large antrum. The theca layers and cumu- lus oophorus may be evident. At the moment, the available data regarding the relation- ship between the MFR and the female reproduction in the offspring have many limitations and gaps concerning the potential underlying mechanisms and almost all have evalu- ated the effects of maternal caloric restriction on the first generation only. Based on these considerations, the aim of the present experimental study was to investigate the impact of MFR in rats on ovarian architecture and function in first and second generations of female offspring. Background They were weighed using a digital bal- ance (0.0001 g) and immediately fixed in 10% neutral buffered formalin at room temperature for classical hist- ology. The remainder of FR1 females were allowed to reach sexual maturity, and were treated exactly as their mothers (FR females), i.e. 50% ad libitum food through- out gestation. After birth, a second generation of the doubly food- restricted group females (FR2) was obtained. The FR2 offspring females were humanely sacrificed at 4 and 8 weeks, and their ovaries were collected, weighed and fixed exactly as before detailed for FR1 offspring females. Number of follicles Total follicles Total number of ovarian follicles was significantly (P = 0.0006) lower in FR1 females at week 4 in comparison with controls, while it was not different in FR2 females when compared to the controls (Fig. 2a). At week 8, the total number of follicles in ovaries from both FR1 and FR2 females resulted significantly (P = 0.0485 and P = 0.0013, respectively) lower than in controls. The total number of follicles was significantly (P = 0.0020 and P = 0.0074, re- spectively) higher and lower in FR2 vs. FR1 females at week 4 and 8, respectively. Antral follicles When compared to controls, the number of antral folli- cles was significantly lower (P = 0.0023) in the ovaries of FR1 females at week 4, and significantly higher (P = 0.0084) in the ovaries of FR2 females at week 8 (Fig. 2e). A significant (P = 0.0008 and P < 0.0001, respectively) difference in the number of antral follicles was detected between FR2 and FR1 females at week 4 and 8. Experimental design First- and second-generation offspring [4, 32] of preg- nant rat exposed directly (or indirectly through germline-independent transmission) to MFR were stud- ied. Specifically, adult virgin Wistar female rats weighing 230 ± 20 g obtained from the Animal Unit at King Saud University were given free access to tap water and were fed ad libitum on standard rodent chow (23% protein, 4.5% fat, 3030 kcal/kg; lab diet 5001, Brentwood, MO). After being maintained in separate cages for four days of adaptation, they were mated with virgin adult males, and then were randomly divided into two groups: control group (group C, n. 15) received ad libitum food, and food-restricted group (group FR, n. 20) received only 50% of ad libitum food throughout gestation. The first A particular interest was given to the occurrence of follicles containing multi-nuclei oocytes (MNOFs), key indicator of perturbations during germ cell nest forma- tion [33–38]. The total number of multi-oocyte follicles (MOFs) was also counted in every section of the ovaries from the different groups. To estimate the number of slides to be read for each ovary we used sample size calculations using the following formula [39]: n≅z2 1‐α=2 s2 h2 where: s = sample standard deviation from an initial number (n0) of Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 3 of 9 week 4 (P = 0.0014 and P < 0.0001, respectively), and at week 8 (P = 0.0002 and P = 0.0044, respectively) (Fig. 2b). week 4 (P = 0.0014 and P < 0.0001, respectively), and at week 8 (P = 0.0002 and P = 0.0044, respectively) (Fig. 2b). replications (n0 = 11), Z1 −α/2 = the value retrieved from the normal standard distribution, corresponding to the 1-α/2 probability (we choose α = 0.05) and h: the half width of the confidence interval. Statistical analysis Statistical analysis For data analysis of follicle number, we used the GraphPad prism version 5. Statistical comparisons were made using a two-tailed t-test. All values are presented as the mean ± standard deviation (SD). Sig- nificance was set at P value less than 0.05. Primordial follicles The amount of the primordial follicle was significantly lower in FR1 and FR2 females than in controls at both Fig. 1 Effect of MFR on ovarian weight results in a significant reduction in 8-week-old FR2 females when compared to controls (P = 0.0003), and to FR1 females (P = 0.0011); no effect is observed in 4-week-old females When compared to the controls, the MOFs were sig- nificantly (P = 0.0044 and P = 0.006, respectively) more frequent in ovaries from FR1 and FR2 females at week 4 (Fig. 2f). Nevertheless, while this number was signifi- cantly higher (P = 0.0013) in FR1 females compared to control at week 8, it was significantly (P = 0.0075) lower in ovaries from FR2 females. Ovarian histopathology MOFs MOFs population was found in all the studied groups and at all follicular stages, from the primordial to the large an- tral stage; these MOFs contained two or more oocytes (Fig. 3a-f). The architecture of the ovaries from FR1 and FR2 females was mainly characterized by more growing follicles when compared to controls. In most of the cases, these were adjacent to each other, suggesting that a fusion has occurred (Fig. 3a). Furthermore, we even reported joining follicles, characterized by the displacement of the oocyte from one follicle into another (Fig. 3b). Ovary weight The number of secondary follicles was significantly (P = 0.0002) lower in the ovaries of FR1 females compared to controls only at week 4 (Fig. 2d). Both at week 4 and 8, the number of secondary follicles was significantly (P = 0.0010 and P = 0.0002, respectively) higher in FR2 than in FR1 females. No difference in mean ovary weight was detected be- tween FR1 and FR2 females vs. controls in 4-week-old. Conversely, in 8-week-old animals a significant (P = 0.0003) difference between intervention and control group in ovarian weight for FR2 females only (Fig. 1). A significant (P = 0.0011) difference was also observed in ovary weight in FR2 vs. FR1 females at week 8 (Fig. 1). Primary follicles The number of primary follicles was significantly (P = 0.034 and 0.003, respectively) higher in FR1 and FR2 vs. controls at week 4. That statistical (P = 0.0072) differ- ences were sustained at week 8 only in FR1 females vs. controls (Fig. 2c). A significant (P = 0.0014) difference in the number of primary follicles was observed in FR2 vs. FR1 females at week 8. MNOFs (Heterokaryon) A high frequency of MNOFs was detected in the ovaries of FR1 and FR2 females compared to controls (Fig. 4a- b). A detailed analysis of these MNOFs provided clues on how they were generated. Specifically, we detected Fig. 1 Effect of MFR on ovarian weight results in a significant reduction in 8-week-old FR2 females when compared to controls (P = 0.0003), and to FR1 females (P = 0.0011); no effect is observed in 4-week-old females Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 4 of 9 Fig. 2 Effect of MFR on the number of follicles per section of ovarian tissue. a Total number of follicles: MFR significantly affects the total number of follicles in ovaries from FR1 females at 4 weeks (P = 0.0006) and 8 weeks (P = 0.0485), and from FR2 females at 8 weeks (P = 0.0013). The total number of follicles from FR2 at week 4 is higher vs. FR females (P = 0.0020), whereas it is lower in FR2 females at week 8 compared to FR females (P = 0.0074). b Primordial follicles: significant effect of MFR on the number of primordial follicles in FR1 and FR2 females at week 4 when compared to control (P = 0.0014 and P < 0.0001, respectively) and in FR2 females at week 8 (P = 0.0002). The total number of primary follicles from FR2 at both week 4 and 8 is significantly lower when compared to FR1 females (P = 0.0020 and P = 0.003, respectively). c Primary follicles are significantly higher in number in ovaries of 4-week-old females in both FR1 and FR2 vs. controls (P = 0.034 and P = 0.003). At week 8, FR1 females have significantly higher number of primary follicles vs. control (P = 0.0072), whereas FR2 females have significantly lower number of primary follicles vs. FR1 (P = 0.0014). d Secondary follicles: FR1 females at week 4 have significantly lower number of secondary follicles vs. control (P = 0.0002) and vs. FR2 (P = 0.001), whereas at week 8 FR2 have significantly higher number of secondary follicles vs. FR1 females (P = 0.0002). e Antral follicles: MFR has the same effect as in secondary follicles, except for a significant increase in FR2 females vs. control at week 8 (P = 0.008). f MOFs: The number of MOFs is significantly higher in ovaries from FR and FR2 females vs. MNOFs (Heterokaryon) control at week 4 (P = 0.0044 and P = 0.006, respectively); it is also significantly higher in FR1 females at week 8 (P = 0.0013), and again significantly lower in ovaries from FR2 females vs. control (P = 0.0075). FR2 females at week 8 have significantly lower number of MOFs when compared to FR1 females (P < 0.0001). All results are given as mean ± SD; P < 0.05, FR1 vs. controls (C) and FR2 vs. controls, with error bars: FR1 vs. FR2 Fig. 2 Effect of MFR on the number of follicles per section of ovarian tissue. a Total number of follicles: MFR significantly affects the total number of follicles in ovaries from FR1 females at 4 weeks (P = 0.0006) and 8 weeks (P = 0.0485), and from FR2 females at 8 weeks (P = 0.0013). The total number of follicles from FR2 at week 4 is higher vs. FR females (P = 0.0020), whereas it is lower in FR2 females at week 8 compared to FR females (P = 0.0074). b Primordial follicles: significant effect of MFR on the number of primordial follicles in FR1 and FR2 females at week 4 when compared to control (P = 0.0014 and P < 0.0001, respectively) and in FR2 females at week 8 (P = 0.0002). The total number of primary follicles from FR2 at both week 4 and 8 is significantly lower when compared to FR1 females (P = 0.0020 and P = 0.003, respectively). c Primary follicles are significantly higher in number in ovaries of 4-week-old females in both FR1 and FR2 vs. controls (P = 0.034 and P = 0.003). At week 8, FR1 females have significantly higher number of primary follicles vs. control (P = 0.0072), whereas FR2 females have significantly lower number of primary follicles vs. FR1 (P = 0.0014). d Secondary follicles: FR1 females at week 4 have significantly lower number of secondary follicles vs. control (P = 0.0002) and vs. FR2 (P = 0.001), whereas at week 8 FR2 have significantly higher number of secondary follicles vs. FR1 females (P = 0.0002). e Antral follicles: MFR has the same effect as in secondary follicles, except for a significant increase in FR2 females vs. control at week 8 (P = 0.008). f MOFs: The number of MOFs is significantly higher in ovaries from FR and FR2 females vs. MNOFs (Heterokaryon) control at week 4 (P = 0.0044 and P = 0.006, respectively); it is also significantly higher in FR1 females at week 8 (P = 0.0013), and again significantly lower in ovaries from FR2 females vs. control (P = 0.0075). FR2 females at week 8 have significantly lower number of MOFs when compared to FR1 females (P < 0.0001). All results are given as mean ± SD; P < 0.05, FR1 vs. controls (C) and FR2 vs. controls, with error bars: FR1 vs. FR2 many MOFs in which oocytes were frequently observed very close to each other (Fig. 4d), suggesting they had fused to form a heterokaryon. In some cases, oocytes within the same follicle were apparently undergoing such a joining process (Fig. 4e). follicles that reaches its maximum level around the time of birth, and is gradually depleted during reproductive life [40–42]. At prepubertal age (week 4), we found that the number of primordial follicles was significantly de- creased in FR1 and FR2 rats, suggesting that MFR affects the ovarian reserve of primordial follicles during early fetal life. The significant decrease in the number of primordial follicles at week 4 was associated with an in- crease in the number of primary follicles among the Discussion The fertile reproductive lifespan of female mammals is mainly linked to the initial ovarian reserve of primordial Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 5 of 9 Fig. 3 (See legend on next page.) Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 6 of 9 (See figure on previous page.) Fig. 3 Hematoxylin and eosin staining of paraffinized ovarian sections from FR1 and FR2 females showing the generation of MOFs. a Ovaries from FR1 and FR2 females are mainly characterized by many growing follicles that are adjacent to each other, indicating their fusion. b-d Follicles in the process of merging (arrowheads); we can see in (d) the displacement of the oocyte from one follicle into the second one. e Primordial follicle with two oocytes. f primordial follicle with three oocytes. g Primary follicle with two oocytes, (h) Primary follicle with three oocytes. i Secondary follicle with two oocytes. (J) Antral follicle with two oocytes; (k) Antral follicle with three oocytes; (l) Antral follicle merging with an early secondary follicle (arrowhead), the large arrow is showing the oocyte position of the antral follicle. Scale bar = 200 μm reproduce successfully [46]. In fact, fetal growth re- striction can be considered as a part of the life his- tory strategy for FR1 and FR2 females that were in utero when their mothers underwent food restriction. Since prenatal undernutrition leads to reduced lon- gevity in mice [47], these females may anticipate a shorter life because of a higher risk of extrinsic mor- tality. It is possible to hypothesize that they may have to adjust their reproductive aptitude by changing the intensity and duration of their lifespan, the timing of the stages of folliculogenesis, as well as the age at which they should reach reproductive maturity. Due to the lack of food sensed through nutritional or endocrine signaling during fetal life [48], and to en- sure reproductive success before death, these females have probably programmed their reproductive lifespan to be very intensive but relatively limited in time, which is consistent with population regulation in the theory of life history [49]. Thus, when they reach different studied groups suggesting that MFR might cause an early menarche by inducing early folliculogen- esis. This hypothesis seems consistent with previous studies [26, 43]. Discussion The rela- tively large number of induced follicles undergoing folliculogenesis at one time makes follicles adjacent to each other and, consequently, highly increases the probability that they will merge to form MOFs [34]. This may have decreased the number of growing oo- cytes (secondary and antral follicles), and may explain the lower total number of follicles, and the higher number of MOFs in 4-week-old FR1 females (vs. con- trols). This strategy is also associated with a faster de- cline in ovarian function with aging, that is clearly supported by the significant decrease in the number of primordial follicles in the ovarian reserve, and also by the significant decrease in the total number of follicles in both FR1 and FR2 females at an early age (8-week), that corresponds in normal females to a high reproductive performance. the assembly of follicles [34] rather than being pro- duced early on during the formation of the ovary. The presence of more than one oocyte in direct con- tact within the same follicle highly increases the pos- sibility of their fusion, and leads to the formation of MNOFs. In fact, any cell brought into contact with another cell and given the right conditions (such as sufficient amounts of fusogen proteins, simultaneously present on each of the two cell surfaces) will fuse with the second cell, even when the latter is foreign [51–53]. This fusion can be beneficial mainly during embryonic development, and for cell-based therapies, and represents a well-known process during reproduction when gametes (spermatozoa and oo- cytes) unite during fertilization to form the zygote. It has also been described in muscle cells, macrophages and nerve cells [54–58]. When cell fusion is blocked during embryonic development, defects in organogen- esis, embryonic lethality, and postembryonic defects can increase [53, 59] suggesting that fused cells are hybrid cells or chimera that function efficiently during embryonic development, driving correct organ forma- tion [54]. In Caenorhabditis elegans, particularly in the proliferative zone of germ cells, two or more crescent shaped nuclei have been observed [60]. MNOFs have also been described during the initial stages of oogenesis in some amphibian species. For example, in Ascaphus truei, oogenesis involves eight nuclei [61], whereas frog oocytes with two nuclei have been described in Leiopelma hochstetteri [62]. Discussion At week 8, the significant decrease in ovarian weight related to the total number of follicles in both first and second generation offspring after MFR is mainly caused by the significant decrease in primordial follicles in the ovarian reserve compared to controls. This could sug- gest an early menopause in FR2, which is less likely in FR1 animals. In fact, previous studies reported that MFR is associated with early menarche and meno- pause [26, 43–45]. To this regard, it is possible to hypothesize that MFR provided first and second gen- eration offspring with a phenotype that is better suited for the lack of food. That new phenotype is consistent with the trade-off theory, i.e. an increase in fertility and a decrease in reproductive lifespan may lead to an increase in the chances of an organism to Fig. 4 Hematoxylin and eosin staining of paraffinized ovarian sections from FR1 and FR2 females showing the generation of MNOFs. a Primordial follicle with two nuclei- oocyte. b Secondary follicle containing one oocyte with two proportional nuclei (c) Secondary follicle containing one merged oocyte with four disproportional nuclei. d Secondary follicle with two oocytes that are very close to each other, suggesting that they will probably fuse soon to form a heterokaryon. e Secondary follicle containing two semi-fused oocytes (arrowhead). f The same secondary follicle in (e) but at another level of section. Scale bar = 200 μm Fig. 4 Hematoxylin and eosin staining of paraffinized ovarian sections from FR1 and FR2 females showing the generation of MNOFs. a Primordial follicle with two nuclei- oocyte. b Secondary follicle containing one oocyte with two proportional nuclei (c) Secondary follicle containing one merged oocyte with four disproportional nuclei. d Secondary follicle with two oocytes that are very close to each other, suggesting that they will probably fuse soon to form a heterokaryon. e Secondary follicle containing two semi-fused oocytes (arrowhead). f The same secondary follicle in (e) but at another level of section. Scale bar = 200 μm Harrath et al. Journal of Ovarian Research (2017) 10:77 Page 7 of 9 Page 7 of 9 prepubertal age, they may upregulate the expression of genes involved in steroidogenesis, which in turn in- duces folliculogenesis for a greater number of primor- dial follicles while concomitantly explaining the significant higher number of in primary follicles in 4- week-old FR1 and FR2 females vs. control. FR1: Female rat offspring from the first generation; FR2: Female rat offspring from the second generation; FSH: Follicle stimulating hormone; H&E: Hematoxylin and eosin; MFR: Maternal food restriction; MNOFs: Follicles containing multi-nuclei oocytes; MOFs: Multi-oocyte follicles; NBF: Neutral buffer formalin; SD: Standard deviation Discussion The most evident example of the necessity for multi- nucleated cells is the syncytiotrophoblast, which is formed when embryonic cytotrophoblast cells fuse with the maternal endometrial epithelium [63, 64]. These cells represent the most important cell type in the placenta, and there is a strong correlation be- tween a successful pregnancy and healthy syncytiotro- phoblasts, likely due to their multi-nuclear state [64]. g p p Of note, a higher number of MOFs in ovaries from 4- week-old first and second generation females was ob- served. Many follicles at these stages may have fused to form MOFs, which explains their significant higher number in FR1 females at four weeks, while this is less likely in FR2 females. While the mechanism of MOFs formation during nest breakdown has been described [33, 50], this is the second study that clearly confirms a new mechanism for the generation of MOFs through the fusion of follicles in the mammalian ovary, and their in- cidence increased sharply at prepubertal age [34]. In- versely, we found that the number of secondary and antral follicles was significantly lower in FR1 and FR2 at prepubertal age when compared to controls. Previous findings have reported that most cases of MOFs repre- sent a fusion between secondary follicles, or a fusion be- tween one secondary and one large antral follicle [34]. Furthermore, the number of secondary/antral follicles observed in 8-week-old females was higher than in con- trols when the number of MOFs was lower. This result is also consistent with the finding of Perez-Sanz and co- workers that showed that the number of MOFs declines significantly in female mice when they become sexually mature [33]. Conclusions Current data suggest that MFR influences ovarian histo- pathology and, in turn, the reproductive health of first and second generation female offspring during fetal de- velopment, and they have been probably programmed to have an early menarche by inducing early folliculogen- esis, and an early decline in ovarian function thereby de- creasing the reproductive lifespan, and leading to an early menopause. The presence of MNOFs is exceptional and repre- sents a challenge for future studies. Two different mechanisms could explain the origin of this phenomenon: the nest breakdown–follicle assembly, and the fusion of more than one oocyte within the same multi-oocyte follicle. Based on our findings, the second mechanism appears the most probable since it is suggested that MOFs are most likely generated by Competing interests 18. Ibanez L, Potau N, Ferrer A, Rodriguez-Hierro F, Marcos MV, De Zegher F. Reduced ovulation rate in adolescent girls born small for gestational age. J Clin Endocr Metab. 2002;87:3391–3. 19. Ibanez L, Potau N, Enriquez G, De Zegher F. Reduced uterine and ovarian size in adolescent girls born small for gestational age. Pediatr Res. 2000;47:575–7. Abbreviations FR1 F l Page 8 of 9 Page 8 of 9 Page 8 of 9 Harrath et al. Journal of Ovarian Research (2017) 10:77 Availability of data and materials There are no shared the data and material for this manuscript. 14. Magee TR, Han G, Cherian B, Khorram O, Ross MG, Desai M. Down- regulation of transcription factor peroxisome proliferator-activated receptor in programmed hepatic lipid dysregulation and inflammation in intrauterine growth-restricted offspring. Am J Obstet Gynecol. 2008;199 Acknowledgements 11. Desai M, Babu J, Ross MG. Programmed metabolic syndrome: prenatal undernutrition and postweaning overnutrition. Am J Physiol-Reg I. 2007;293: R2306–14. We would like to thank Dr. Deborah Sloboda from the Departments of Biochemistry and Biomedical Sciences, Obstetrics and Gynecology, and Pediatrics, McMaster University, Hamilton, Canada, for the constructive criticism on an earlier draft of the manuscript. 12. Bieswal F, Ahn MT, Reusens B, Holvoet P, Raes M, Rees WD, Remacle C. The importance of catch-up growth after early malnutrition for the programming of obesity in male rat. Obesity. 2006;14:1330–43. Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. 20. Cooper C, Kuh D, Egger P, Wadsworth M, Barker D. Childhood growth and age at menarche. Brit J Obstet Gynaec. 1996;103:814–7. 20. Cooper C, Kuh D, Egger P, Wadsworth M, Barker D. Childh age at menarche. Brit J Obstet Gynaec. 1996;103:814–7. 21. Veening MA, van Weissenbruch MM, Roord JJ, Delemarre-van de Waal HA: Pubertal development in children born small for gestational age. J Pediatr Endocr Met 2004, 17:1497–1505. Funding 13. Thompson NM, Norman AM, Donkin SS, Shankar RR, Vickers MH, Miles JL, Breier BH. Prenatal and postnatal pathways to obesity: different underlying mechanisms, different metabolic outcomes. Endocrinology. 2007;148: 2345–54. The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for funding this research RG-164. The authors would like to extend their sincere appreciation to the Deanship of Scientific Research at King Saud University for funding this research RG-164. Availability of data and materials There are no shared the data and material for this manuscript. Author details 1 1Zoology Department, College of Science, King Saud University, Riyadh, Saudi Arabia. 2Unit of Reproductive and Developmental Biology, Faculty of Science of Tunis University of Tunis El Manar, Tunis, Tunisia. 3Unit of Gynecology and Obstetrics, Grande Ospedale Metropolitano “Bianchi – Melacrino – Morelli”, Reggio Calabria, Italy. 22. Ibanez L, de Zegher F. Puberty and prenatal growth. Mol Cell Endocrinol. 2006;254:22–5. 23. 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https://openalex.org/W2805392662	https://hal.archives-ouvertes.fr/hal-02324246/document	English	null	When the Sad Past Is Left: The Mental Metaphors Between Time, Valence, and Space	Frontiers in psychology	2,018	cc-by	14,204	1In this study, we use the terminology “mental metaphor” as termed by Casasanto (2013) to refer to what Lakoﬀand Johnson (1980) ﬁrstly called “Conceptual Metaphor.” The reasons for this change is that the terminology “conceptual metaphor” refers to mental metaphors being stored within the structure of the borrowing concept, in long term memory, and to a metaphoric process used exclusively to help the processing of abstract concepts, two assumptions that we are rejecting in the current article. Edited by: Edited by: Pia Knoeferle, Humboldt-Universität zu Berlin, Germany Reviewed by: Laura J. Speed, Radboud University Nijmegen, Netherlands Fernando Marmolejo-Ramos, University of Adelaide, Australia Michele Burigo, Bielefeld University, Germany Correspondence: Marc Ouellet mouellet@ugr.es Specialty section: This article was submitted to Language Sciences, a section of the journal Frontiers in Psychology When the Sad Past Is Left: The Mental Metaphors Between Time, Valence, and Space Nicolas Spatola 1, Julio Santiago 2,3, Brice Beffara 4,5,6, Martial Mermillod 4,5, Ludovic Ferrand 1 and Marc Ouellet 1,2,3 1 Université Clermont Auvergne, CNRS, LAPSCO, Clermont-Ferrand, France, 2 Department of Experimental Psychology, University of Granada, Granada, Spain, 3 Center for Research on Mind, Brain, and Behavior (CIMCYC), University of Granada, Granada, Spain, 4 Univ. Grenoble Alpes, CNRS, LPNC, Grenoble, France, 5 Univ. Grenoble Alpes, LJK, Grenoble, France, 6 Psychological Sciences Research Institute, Université Catholique de Louvain, Louvain-la-Neuve, Belgium A mental metaphor is a strategy that consists of completing the representation of a concept with structural components of a correlating concept. Three issues were addressed here to deepen our understanding of this mechanism: the use of mental metaphors between abstract concepts, the simultaneous activation of multiple mental metaphors and the importance of the focus of attention on the relevant dimensions of a mental metaphor. In two experiments, participants made temporal or valence judgments (with their left or right hand) on verbs with a negative or positive meaning and conjugated in the past or future form, allowing for the simultaneous activation of the “time is space”, “valence is space,” and “time is valence” mental metaphors. Left-past/right-future and left-negative/right-positive congruency effects were found, and these effects were greater in the temporal and valence judgment tasks, respectively, demonstrating the importance of attentional cuing. Simultaneously, a congruency effect between the abstract concepts of time and valence (past-negative/future-positive) was observed, revealing that a mental metaphor can occur between abstract concepts and that multiple metaphors can be processed simultaneously. These results are discussed in terms of different theories within the ﬁeld of mental metaphors. Keywords: mental metaphor, conceptual metaphor, coherent working models, time, valence, space ORIGINAL RESEARCH published: 28 June 2018 doi: 10.3389/fpsyg.2018.01019 INTRODUCTION Received: 07 November 2017 Accepted: 31 May 2018 Published: 28 June 2018 Research on conceptualization has demonstrated that participants frequently resort to a metaphorical strategy when processing a concept. This strategy consists of borrowing structural components of a correlating concept to complete the structure of the concept at hand. A good example of this phenomenon is the well-studied mental metaphor1 of time onto the sagittal axis Citation: Spatola N, Santiago J, Beffara B, Mermillod M, Ferrand L and Ouellet M (2018) When the Sad Past Is Left: The Mental Metaphors Between Time, Valence, and Space. Front. Psychol. 9:1019. doi: 10.3389/fpsyg.2018.01019 June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org The Sad Past Is Left Spatola et al. that can be quantiﬁed as more or less, such as quantity, space, time or even valence) share neural substrates to process the dimension of magnitude. The simultaneous processing of concepts of magnitude would automatically lead to congruency eﬀects because the diﬀerent dimensions of magnitude would be processed as a consequence of a shared neural mechanism. A further proposal, the Coherent Working Models (Santiago et al., 2011; for a similar view, see also Marmolejo-Ramos et al., in press), suggests that the mental metaphors are “built” in working memory while the task is carried out. According to the Coherent Working Models, a mental model of the task would be built in working memory to meet task goals. This mental model would include the concepts that are correlating within a task (such as time and space when walking through a path) and form a common representation that is coherent with task goals (past- behind/future-front, for the previous example). After repeated use, a mental metaphor could be stored in long-term memory for further use, but not within the structure of the borrowing concept. This mental metaphor would need to be reactivated in working memory in order to again be part of a mental model. These two alternative explanations have in common the notion that the degree of abstractness (or concreteness) of a concept is no more an issue and they predict bi-directional inﬂuences between the concrete and the abstract domains. of space (past-behind/future-front). This mental metaphor would be a mental transmutation of the experience of time ﬂowing while walking through a path, leaving the past behind and moving forward toward the future (Lakoﬀand Johnson, 1999). The result, according to Santiago et al. (2011), is a more eﬃcient conceptual representation for the task that is to be carried out with the concept. Even though the mechanisms involved in this metaphoric process have received much attention during the past decades, many issues are still unresolved or are a matter of debate. Can the Processing of an Abstract Concept Modulate the Processing of Another Abstract Concept? Accordingly, Zhong and Liljenquist (2006), Zhong and Leonardelli (2008) and Ouellet et al. (2010a) have demonstrated that processing information from the abstract domain can modulate the processing of the information of the concrete domain. First, Zhong and Liljenquist (2006) studied the link between the purity of the body and morality. Participants were asked to recall an unethical or an ethical deed. They demonstrated that the participants who recalled an unethical deed were more prone to take antiseptic wipes than the participants who recalled an ethical deed. Second, Zhong and Leonardelli (2008) studied the link between the domain of temperature and that of social relationships. People can speak of a “cold atmosphere” or a “warm person” when speaking about social relationships. In their study, Zhong and Leonardelli (2008) demonstrated the psychological reality of this metaphor by asking their participants to recall a social exclusion or a social inclusion experience. This mere recall was enough to modulate the perception of the temperature in the room. Participants who recalled a social exclusion experience estimated the temperature of the room to be lower than those who recalled a social inclusion experience. Third, Ouellet et al. (2010a) used temporal stimuli to orient participants’ spatial attention. They observed a congruency eﬀect according to the left-past/right-future mental metaphor. Based on linguistic evidence, Lakoﬀand Johnson (1980) were the ﬁrst to systematically study mental metaphors, adopting the view that they termed Conceptual Metaphor Theory. Their inventory of linguistic expressions reﬂecting this structural borrowing process was mainly composed of abstract concepts beneﬁting from the structures of concrete concepts. For example, in the expressions “I am looking forward to their response” or “I am feeling up today,” the concept of space is used to understand the concepts of time and valence, respectively. On the opposite side, there are no or very few expressions of space being understood by the concepts of time or valence (e.g., you cannot say “I was looking at the plane positive in the sky” or “John sat in the future of me”). Lakoﬀand Johnson (1980) concluded that abstract concepts must have a structure that is not as complete as that of concrete concepts due to a lack of grounding in sensory-motor experiences. In order to complete the content of an abstract concept, elements from a more complete conceptual structure (the structure of a concrete concept) would be borrowed. Citation: This study aims to address three important issues in the ﬁeld of mental metaphors in order to respond to the following questions: (1) “Can the processing of an abstract concept modulate the processing of another abstract concept?”; (2) “Can multiple mental metaphors be processed at the same time and, if this is the case, what are the underlying mechanisms allowing for this simultaneous activation?”; (3) “Is the focus of attention important for activating a mental metaphor?” Each issue is discussed in detail below. Frontiers in Psychology \| www.frontiersin.org June 2018 \| Volume 9 \| Article 1019 Can the Processing of an Abstract Concept Modulate the Processing of Another Abstract Concept? The results of previous studies investigating the psychological reality of Conceptual Metaphor Theory supported this central assumption. One of the most famous demonstrations was that by Boroditsky (2000), who studied an eﬀect resulting from the mental metaphor of time onto the sagittal spatial axis. Boroditsky (2000) observed that the processing of the concrete domain of space could modulate the processing of the abstract domain of time, but not the other way around. According to her, the frequent use of a mental metaphor would lead to its storage in long-term memory, within the structure of the abstract concept. As a result, the activation of the abstract concept, but not the concrete concept, would also activate the mental metaphor. Moreover, Santiago et al. (2012) were able to observe a bi-directional conceptual congruency eﬀect for the negative- down/positive-up mental metaphor. In their study, participants were asked to make valence (Experiments 2 and 3) or spatial (Experiments 4 and 7) judgments on positive or negative emotional words presented above or below a face silhouette. When the instructions were simply to make valence (Experiment 2) or spatial (Experiment 4) judgments, they did not observe any congruency eﬀect between the dimensions of valence and space. However, when the participants were endogenously cued to the However, alternative explanations have been proposed. Walsh (2003) put forward the ATOM theory (A Theory Of Magnitude). According to this theory, all concepts of magnitude (concepts June 2018 \| Volume 9 \| Article 1019 2 The Sad Past Is Left Spatola et al. would pay for a bottle of wine in order to thank a friend for letting them use his vacation home... 1 week ago or in 1 week from now. Caruso et al. (2008) demonstrated that people assigned a diﬀerent monetary value to similar events depending on whether they were past or future events. Guo et al. (2012) also demonstrated that this eﬀect was linked to cultural factors; whilst European Canadians gave more value to events in the future than to events in the past, Chinese Canadians showed the opposite pattern. Guo et al. (2012) attributed this cultural eﬀect to the diﬀerent ways of representing time. In Chinese culture, time is perceived as something cyclic, where the past is central because the future is a simple repetition of what has already happened (Biao, 2001; Guo et al., 2012). Can the Processing of an Abstract Concept Modulate the Processing of Another Abstract Concept? In contrast, North Americans have a more linear representation of time in which the future is more important because it is subject to a certain level of control, whereas the past is seen as something that cannot be changed (Graham, 1981; Spears et al., 2001). However, studies on the TVA eﬀect cannot be considered as deﬁnitive proof of a congruency eﬀect between two abstract concepts. Eﬀectively, the valuation is made via the use of a concrete concept (money) and the reference to the past or the future is made via reference to episodic events, which are much more concrete than the reference to the past or the future in general. As a consequence, the TVA congruency eﬀect could be explained by an association between the concrete concepts of money and episodic events, rather than an association between the abstract concepts of time and valence. irrelevant dimension of the task (by asking them to report in which position more words appeared in the valence judgment task or whether there were more positive or negative words in the spatial judgment task), they observed a congruency eﬀect between the dimensions of valence and space in both tasks. Eﬀectively, apart from observing that the concrete spatial domain of space could modulate the processing of valence (Experiment 3), they also observed that the abstract domain of valence could modulate the processing of space (Experiment 7). Thus, the key factor was the attentional focus given to the irrelevant conceptual dimension of the task, rather than the degree of abstractness (or concreteness) of the prime vs. the target. Another study by Rusconi et al. (2006) also demonstrated that a mental metaphor is possible between two concepts with a low level of abstractness. Eﬀectively, they observed a congruency eﬀect between space and pitch. In their study, when participants were asked to make discrimination judgments on high- and low- pitched sounds with an upper or a lower response key, they were better at making “up” and “down” responses for high- frequency and low-frequencies pitches sounds, respectively, than the opposite. The last piece missing in order to discard the claim that a mental metaphor necessarily involves an abstract concept beneﬁting from the more complete structure of a concrete concept is to demonstrate that a mental metaphor can occur between two concepts with a high level of abstractness. Can the Processing of an Abstract Concept Modulate the Processing of Another Abstract Concept? Any interaction between valence and time concepts under these circumstances could only be explained by an interaction between concepts with a high level of abstraction. Can the Processing of an Abstract Concept Modulate the Processing of Another Abstract Concept? To the best of our knowledge, this type of mental metaphor has not yet been demonstrated. From our point of view, the studies that have come closest to achieving this are those showing that the perception of short temporal durations can be modulated by the processing of an abstract concept such as numbers (participants perceive the duration of a stimulus to be shorter when processing low numbers and longer when processing high numbers; see Dormal et al., 2006; Roitman et al., 2007; Vicario et al., 2008). However, a short duration perceived in an online task may not qualify as a concept with a high level of abstractness, due to the fact that the perception of short durations can be directly perceived thanks to the ticking of the participants’ internal clock, as discussed by most of the authors of these studies. Even if the ﬁndings of Guo et al.’s (2012) study do not constitute direct evidence for the positive and negative valuations of the past and future in a more general and abstract way, it can help to predict the congruency eﬀect that should be obtained with French participants, who also seem to represent time in a linear fashion (Yamada and Kato, 2001). Following this logic, French participants should show a past-negative/future- positive congruency eﬀect. It is also important to note that the task used by Guo et al. (2012) caused the participants to implement a reﬂective process. Consequently, it is possible that the mechanisms used to value temporal concepts under these task conditions might diﬀer from those (possibly more automatic processes) measured in the mental metaphor literature. For this reason, the task used here was developed to measure more automatic aspects of the time-valence mental metaphor. In the present study, two concepts with a high level of abstractness were used—time and valence. In order to ensure the abstract nature of the concept of time, verbs conjugated in either the past or future tense were used to refer to these two poles of the time concept (for other studies using a similar strategy, see Torralbo et al., 2006; Ouellet et al., 2010a,b). The second abstract dimension was that of valence (positive or negative). Both dimensions were simultaneously activated due to the use of French positive or negative verbs conjugated in past or future forms (e.g., “haïssaient”—“they hated” or “applaudira”— “he will applaud”). Frontiers in Psychology \| www.frontiersin.org Can Multiple Mental Metaphors Be Processed at the Same Time and, If This Is the Case, What Are the Underlying Mechanisms Allowing for This Simultaneous Activation? Vicario et al. (2008) studied the simultaneous activation of two mental metaphors: the “short durations—low numbers/long durations—high numbers” and the “short durations—left/long durations—right” mental metaphors. Their ﬁndings suggest that at least two mental metaphors can be used at the same time; while participants were making underestimations on the left side and overestimations on the right side they were also making underestimations and overestimations with low and high numbers, respectively. Unfortunately, the analyses conducted Studies on the temporal value asymmetry (TVA) point to the existence of interactions between past-future time and positive- negative valence (e.g., Caruso et al., 2008; Guo et al., 2012). In these studies participants are asked to economically value past or future events. For example, they can be asked how much they June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 3 The Sad Past Is Left Spatola et al. by Vicario et al. (2008) did not allow for an assessment of whether these simultaneous activations were independent or whether they interacted together to form a kind of conglomerate representation. Instead of using statistical methods that would have included all the factors (Durations, Space, and Numbers) in a unique design, which allows for conﬁrming the absence or presence of a three-way interaction, they carried out separated analyses for the congruency eﬀects between Durations and Numbers, and between Durations and Space. (negative or positive) judgments with their left or right hand on negative or positive verbs conjugated to the past or the future would allow the simultaneous activation of three mental metaphors at the same time. Independently of the interacting or independent activation of the mental metaphors, French right-handed participants should show negative-left/positive- right (Valence-Space), past-left/future-right (Time-Space) and past-negative/future-positive (Time-Valence) congruency eﬀects. It is of importance to examine whether the simultaneous activations of multiple mental metaphors are independent or dependent of each other, since this can provide insights into the nature of the mechanisms that allow for their corresponding congruency eﬀects. As explained above, Boroditsky (2000) considered the possibility that the mental metaphors are stored in long-term memory within the representation of the concepts that beneﬁt from the structural borrowing process. According to her, processing such a concept would automatically activate its associated mental metaphor. In addition, when a mental metaphor is activated, it could not be manipulated. Is the Focus of Attention Important for Activating a Mental Metaphor? Activating a Mental Metaphor? Santiago et al. (2011), based on an extensive review of the literature, concluded that the relevant concepts of a mental metaphor needed to be activated to a certain degree for a congruency eﬀect to happen. It thus follows that attentional factors should play a key role in this conceptual activation. Santiago et al. (2012) were the ﬁrst to systematically investigate how the focus of attention on the relevant concepts of a mental metaphor could modulate its activation. As explained above, the type of judgment (on valence or vertical space) oriented participants’ attention toward the concept that had to be judged and, in order to observe a positive-up/negative-down congruency eﬀect, attention also had to be cued to the irrelevant concept of the task. Another ﬁnding is that of Torralbo et al. (2006) who studied a concept (time) that can be mapped onto various conceptual dimensions (sagittal or coronal spatial axis). In their study, participants were asked to judge either verbally or manually (with their left and right hands) the temporal reference of words appearing to the left or right on the screen. The left-past/right-future congruency eﬀect emerged only in the task with manual judgments. The use of the left and right hands cued participants’ attention to these spatial locations and allowed the mapping of time onto the left-right horizontal axis. In the same vein, de la Vega et al. (2012) demonstrated that asking right-handed participants to perform a simple lexical judgment task (Experiment 1) on positive and negative words with their left and right hands was not enough to observe a left- negative/right-positive congruency eﬀect. The activation of this mental metaphor was observed only when the attention of the participant was cued on the valence dimension of the stimuli by asking them to make positive or negative judgments on these stimuli (Experiment 2). In their last experiment (Experiment 4), they also demonstrated that attention needed to be cued on the spatial dimension as well by showing that the congruency eﬀect emerged only when the mapping of the spatial response was explicit, not implicit. Another approach (Santiago et al., 2011; see also Marmolejo- Ramos et al., in press) suggests that the conceptual maps are “built” in working memory while the task is carried out and/or depend on the simultaneous use of the same neural substrates (Walsh, 2003). Can Multiple Mental Metaphors Be Processed at the Same Time and, If This Is the Case, What Are the Underlying Mechanisms Allowing for This Simultaneous Activation? In this case, varying the degree of activation of the conceptual components of a mental metaphor would not have any eﬀect on the processing of that mental metaphor. Thus, processing one mental metaphor that has a particular conceptual component could not modulate the simultaneous processing of another mental metaphor that also has this same conceptual component. For example, if a participant reads the verb “they will succeed”, she/he would activate the “left-negative/right-positive” and the “left- past/right-future” mental metaphors. Both metaphors—activated by this stimulus—would prime the space on the right, but the congruency eﬀect observed for one mental metaphor would not interact with the congruency eﬀect of the other mental metaphor. In other words, the congruency eﬀects would be additive. Frontiers in Psychology \| www.frontiersin.org June 2018 \| Volume 9 \| Article 1019 Is the Focus of Attention Important for Activating a Mental Metaphor? In both cases (Walsh, 2003; Santiago et al., 2011), the structural components participating in the distinct mental metaphors would be activated and simultaneously used to form these metaphors. Thus, two or more mental metaphors sharing a conceptual domain would also share common resources and compete for the simultaneous use of these resources. As a result, each mental metaphor would modulate the processing of the other, and the congruency eﬀects should therefore interact. Time and Valence concepts are particularly interesting because they share a common spatial projection on the left- right horizontal axis. On the one hand, the past-left/future-right mental metaphor of time would originate from the correlation between the directional activity of reading/writing and the passage of time during this activity (Ouellet et al., 2010b). On the other hand, Casasanto and Chrysikou (2011), and de la Vega et al. (2013; see also de la Vega et al., 2012, Experiments 2 and 3) demonstrated that the negative-left/positive-right mental metaphor of valence depends on the greater ﬂuency experienced on the dominant side of the body. Consequently, asking participants to make temporal (past or future) or valence Based on these ﬁndings, in the present study participants should show greater congruency eﬀects for the mental metaphors that have their conceptual components cued by the task. Because participants had to make temporal and valence judgments (with their left and right hands) in separate blocks, it was expected that the past-left/future-right and negative-left/positive-right congruency eﬀects would be greater in the temporal and valence judgment tasks, respectively. Because the relative salience of the irrelevant conceptual dimension for the past-negative/future-positive congruency Frontiers in Psychology \| www.frontiersin.org June 2018 \| Volume 9 \| Article 1019 4 The Sad Past Is Left Spatola et al. eﬀects was similar in both tasks, it was anticipated that this congruency eﬀect would be of a similar size in both tasks. greater in the temporal compared to the valence judgment task whilst the converse should be true for the negative-left/positive- right congruency eﬀect. With respect to the past-negative/future- positive congruency eﬀect, we expected this to be of a similar size in both judgment tasks because the salience of the irrelevant dimension in each task was similar. If attention does not modulate the activation of the mental metaphors used here, it would imply that their activation is more automatic than originally hypothesized by Santiago et al. (2011). Material The stimuli were 20 positive and 20 negative verbs (see Appendix I for a list of the materials). The positive or negative value of each verb was ﬁrst determined on the intuition of the experimenters and a post-test with 12 French native speakers unaware of the goals of the present study served to test whether or not these sets of verbs diﬀered signiﬁcantly in their ratings. The judgment task was programmed in E-Prime and each verb appeared once in a random order at the inﬁnitive form. A Likert scale was used (1: Extremely negative; 2: Very negative; 3: Negative; 4: Neutral; 5: Positive; 6: Very positive; 7: Extremely positive) and, for each verb, the judge had to press one of the keys from 1 to 7. The mean values for the positive and negative verbs were 5.85 (SE = 0.28) and 2.38 (SE = 0.25), respectively (the mean scores for each verb are presented in Appendix I). This diﬀerence was signiﬁcant in the analyses by participants [t1(11) = −30.008, p < 0.001] and by items [t2(38) = −34.453, p < 0.001]. Second, we predicted that the mental metaphors between time and valence, time and the coronal axis, and valence and the coronal axis should be activated simultaneously. Looking at whether these simultaneous activations occur independently one of each other or whether they interact will deepen our understanding of the mechanism that allows these simultaneous activations. Independent activations would suggest that mental metaphors are stored in long-term memory, within the structure of the concept that beneﬁts from the lending process (Boroditsky, 2000). These representations would be ﬁxed and could not be manipulated. Interactions between these activations would suggest that mental metaphors are elaborated during the accomplishment of the task. Interactions would occur because the same resources would be used by multiple metaphors at the same time. While the Coherent Working Models (Santiago et al., 2011; see also Marmolejo-Ramos et al., in press) would explain such a phenomenon in terms of the simultaneous use of conceptual information in working memory, the ATOM theory (Walsh, 2003) would suppose that this is due to the shared use of neural substrates to process the domain of magnitude. Two lists of 80 stimuli were created. Participants The participants were 32 (28 women; mean age = 18.88 years, SD = 1.17) undergraduate students. They were all from the University Blaise Pascal of Clermont-Ferrand (France) and took part in this experiment in exchange for course credit. All participants were French native speakers and reported having normal or corrected to normal vision. They all gave informed written consent according to the Declaration of Helsinki. Is the Focus of Attention Important for Activating a Mental Metaphor? These predictions run counter to Boroditsky’s (2000) and Walsh’s (2003) theories. As explained earlier, Boroditsky (2000) assumed that, with time, a mental metaphor becomes a structural component of the borrowing concept. Consequently, the borrowing concept cannot be processed without the activation of its associated mental metaphors. Thus, the degree of attention allocated to the lending concept could not modulate the degree of activation of a mental metaphor. Regarding Walsh’s (2003) theory, the activation of the magnitude domain would also be automatic when processing a concept that can be quantiﬁed. Thus, attention could not modulate the congruency eﬀects between these concepts. Frontiers in Psychology \| www.frontiersin.org HYPOTHESES First, based on the ﬁndings that mental metaphors can exist between concepts at similar levels of abstractness (Rusconi et al., 2006) and the fact that people from western cultures tend to value more positively future than past events (Caruso et al., 2008; Guo et al., 2012), we expected that the mental metaphor between the abstract dimensions of time and valence would occur so that a past-negative/future-positive congruency eﬀect should emerge. In particular, negative verbs conjugated to past and positive verbs conjugated to the future should be responded to more rapidly and more accurately than negative verbs conjugated to future and positive verbs conjugated to the past, respectively. Design A 2 (Type of judgment: Time vs. Valence) × 2 (Time: Past vs. Future) × 2 (Valence: Negative vs. Positive) × 2 (Response side: Left vs. Right) within-subject design was used. Material In a ﬁrst list, half of the verbs were presented inﬂected to the 3rd singular person of the past and to the 3rd plural person of the future, and the other half was presented to the 3rd plural person of the past and to the 3rd singular person of the future (see Table 1 for examples of the stimuli). The same verbs were presented in a second list, but with the opposite conjugation arrangement. Special care was taken to ensure the consistency of syllable structure, word length, and frequency of the verb lemma across all conditions (see Table 2). All characters were written in lower case, bold Courier font, point size 18 and presented on a computer screen on a light gray background. Third, based on previous results, we also predicted that the focus of attention should play a role in the activation of the mental metaphors. Because the temporal dimension will be cued in the temporal judgment task (with the left and right hands), the past-left/future-right congruency eﬀect should be The task was programmed in E-prime and ran on an Intel Core 2 Duo 1.8 GHz computer. The stimuli were presented on a June 2018 \| Volume 9 \| Article 1019 5 The Sad Past Is Left Spatola et al. TABLE 1 \| Examples of the stimuli as a function of their valence, temporal reference, and number. Temporal reference Number Valence Negative Positive Past Singular “Trahissait—“she/he betrayed” “Réussissait”—“she/he succeeded” Plural “Trahissaient”—“they betrayed” “Réussissaient”—“they succeeded” Future Singular “Trahira”—“she/he will betray” “Réussira”—“she/he will succeed” Plural “Trahiront”—“they will betray” “Réussiront”—“they will succeed” TABLE 1 \| Examples of the stimuli as a function of their valence, temporal reference, and number. TABLE 2 \| Means of the principal characteristics of the stimuli (Number of letters, Number of syllables, Frequencies of occurrence in texts and movies data bases) and Standard Errors (in brackets) as a function of the Valence, Temporal reference, and Number of the verbs. (temporal or valence). Each main block was further divided into two sub-blocks. Within each sub-block, the complete list of verbs was presented. These sub-blocks diﬀered in terms of the mapping of left and right responses for “past” or “future” judgment in the temporal judgments task, and in terms of the mapping of left and right responses for “negative” or “positive” judgments in the valence judgments task. Material In other words, each participant had to carry out both valence and temporal tasks once with a congruent and once with an incongruent response-key assignment, in counterbalanced order. TABLE 2 \| Means of the principal characteristics of the stimuli (Number of letters, Number of syllables, Frequencies of occurrence in texts and movies data bases) and Standard Errors (in brackets) as a function of the Valence, Temporal reference, and Number of the verbs. Temporal reference Number No. of letters and syllables Valence Negative Positive Past Singular No. of letters 9.7(0.4) 10(0.4) No. of syllables 3.1(0.1) 3.1(0.2) Plural No. of letters 11.7(0.4) 12(0.4) No. of syllables 3.1(0.1) 3.1(0.2) Future Singular No. of letters 7.6(0.3) 7.8(0.3) No. of syllables 3.1(0.1) 3.2(0.2) Plural No. of letters 9.6(0.3) 9.8(0.3) No. of syllables 3.1(0.1) 3.2(0.2) Frequencies of occurrences Texts 69.1(27.3) 86.7(31) Movies 46.7(18.3) 64.1(15.1) The frequencies of occurrences of the verb lemma are expressed in occurrences per million and are from a texts database and a movies database (New et al., 2004). Results are rounded to the nearest tenth. The order of presentation of the two types of judgment was counterbalanced across participants, as was the order of presentation of the blocks diﬀering in response mapping, leading to a total of eight diﬀerent levels of counterbalancing. The complete sequence of the experiment was repeated twice. The trial structure was as follows (see Figure 1). First, a ﬁxation cross (a plus sign) was presented at the center of the screen for 500 ms. The target then appeared for 4,000 ms or until a response was made. Finally, there was a blank screen for 1,500 ms before the next trial. If the participant made an error or did not respond to the target, a BEEP was sounded following the removal of the target. The frequencies of occurrences of the verb lemma are expressed in occurrences per million and are from a texts database and a movies database (New et al., 2004). Results are rounded to the nearest tenth. When the experiment was ﬁnished, the French version of the HAD questionnaire was presented to the participant. The participant was instructed to ﬁll it in as honestly as possible and to place it in a box when ﬁnished, to ensure anonymity. 20-inch LG Flatron screen (6 ms refreshing rate), and an E-prime SRBOX was used to collect participants’ responses. 2The results corresponding to HAD questionnaire will be analyzed in a further study that will include the results of a greater number of participants in order test whether or not the participants present conceptual biases associated with their mood. Results The data set is deposited in a public repository (Spatola et al., 2017). The data from two participants were discarded because they responded randomly in at least one block (around 50% of accurate responses). Responses to two verbs (“vaincre,” meaning “to win,” and “évanouir,” meaning “to faint”) in all conjugated forms were also removed from the analyses because errors were made on more than 25% of the trials in the valence judgment task. The results obtained from the remaining participants and items are summarized in Appendix II. Errors occurred on 3.41% (601 trials) of the trials and these trials were excluded from the reaction times analyses. Correct trials with a reaction time lower or higher than 2.5 SD (standard deviations) per condition for each participant were considered outliers (this ﬁltering procedure has the advantage of taking out extreme values without aﬀecting the data of one condition or of one participant in particular) and also excluded from the reaction time analyses, which corresponded to 583 trials (3.42% of the trials). In order to ensure that the observed eﬀects were not due to the type Material The French version of the hospital anxiety and depression scale (HAD) was used to assess the participants’ anxiety and depression levels (Razavi et al., 1989)2. Procedure and Design Participants sat in a quiet room at approximately 60 cm from the screen. They were ﬁrst informed that they would have to make either temporal (past or future) or valence (negative or positive) judgments on verbs in diﬀerent blocks. They were also instructed that they would have to perform these judgments as fast and accurately as possible with their left and right indexes on the left- most and right-most buttons (7.8 cm form center to center) of the SRBOX, respectively. The ﬁrst list of verbs was assigned to half of the participants and the second list to the other half. The experiment was divided into two main blocks, each one dedicated to one type of judgment June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 6 Spatola et al. The Sad Past Is Left FIGURE 1 \| Trial structure in Experiment 1 and Experiment 2. “trahira” means “she/he will betray”. FIGURE 1 \| Trial structure in Experiment 1 and Experiment 2. “trahira” means “she/he will betray”. backward elimination can be summarized as follows. In their study, Barr et al. (2013) have shown that the best models were those that use maximal random eﬀect structures justiﬁed by the design, followed by the models that use a backward elimination procedure (starting with the maximal structure). Both methods of data analysis have proved to be eﬃcient at avoiding Type I error. However, Bates et al.’s (2015) have demonstrated that Barr et al. (2013) ﬁndings were diﬃcult to apply to real data. They demonstrated that maximal random eﬀect structures tend to not converge and that using too many parameters can lead to models that are diﬃcult to interpret. Thus, in order to have a model that is adequate for our experimental design and to avoid Type I error as much as possible, we determined the maximal random eﬀects structure justiﬁed by the design before using the backward elimination procedure. of ﬁlter used, a multiverse analysis was carried out (Steegen et al., 2016). The reaction time ﬁlters compared to the one we chose were: 2 median absolute deviations (MAD) and 2 SD per condition for each participant, and no reaction time ﬁlter (see Marmolejo-Ramos et al., 2015, for a discussion of the most appropriate reaction time ﬁlters according to the characteristics of the data; see also Marmolejo-Ramos et al., 2015). Procedure and Design We did not use a logarithmic transformation, because it is problematic when one condition is signiﬁcantly slower than the other (Lo and Andrews, 2015); judgments of valence were much slower than temporal judgements in the present study. We did not observe any signiﬁcant interaction between the use of the diﬀerent ﬁlters and the eﬀects of core interest for the present study. The pattern was always similar, but excess data were removed with ±2 SD and ±2 MAD (>5%) and the results were somewhat noisier without any ﬁlter. Only the signiﬁcant or marginally signiﬁcant ﬁndings (with an alpha level < 0.1) corresponding to these models are reported in the text. The results of core interest are summarized in Table 2. To assess the eﬀects of Type of judgment (Temporal vs. Valence), Time (Past vs. Future), Valence (negative vs. positive) and Response side (left vs. right) on latency and accuracy data, separate linear mixed models (LMM) were used, one for each dependent variable Brysbaert, 2007; Baayen et al., 2008; Bates et al., 2015). Analyses were run with the lmerTest package for R (Kuznetsova et al., 2016; R Core Team, 2017). Backward elimination of non-signiﬁcant eﬀects of the linear mixed eﬀect model was performed with the step function. The p-values for the ﬁxed eﬀects were calculated based on Sattethwaite’s approximation. The model included Type of judgment, Time, Valence and Response side as Fixed eﬀects factors, Subjects and Items as Random eﬀects factors, and the by-subject random slopes for valence (using R syntax, the model tested for the reaction times results was: RT ∼Type_of_judgment ∗Time ∗ Valence ∗Response_side + (1+Valence\|Subjects) + (1\|Items)). The rationale behind the choice of this model and the use of Accuracy Analyses Accuracy analyses revealed a signiﬁcant interaction between Valence and Response side factors [b = 2.54, t(9,014) = 3.132, p < 0.01]. What is more, this interaction was modulated by the Type of judgment factor [b = −3.42, t(9,014) = −2.978, p < 0.01; see Figure 2]. The three-way interaction between Type of judgment, Time and Response side was also signiﬁcant [b = 2.46, t(9,014) = 2.138, p < 0.05; see Figure 3]. The nature of these three-way interactions was further examined by conducting separate multilevel models on each level of the Type of judgment factor, including both the two-way interactions of Valence by Response side, and Time by Response side. While the interaction between Valence and Response side was signiﬁcant in valence judgment task [b = 2.55, t(4,460) = 2.966, p < 0.01], but not in the June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 7 Spatola et al. The Sad Past Is Left FIGURE 2 \| Percentage of correct trials per condition in Experiment 1, as a function of Type of judgment, Valence, and Response side. p < 0.01. FIGURE 2 \| Percentage of correct trials per condition in Experiment 1, as a function of Type of judgment, Valence, and Response side. p < 0.01. orrect trials per condition in Experiment 1, as a function of Type of judgment, Valence, and Response side. *p < 0.01. temporal judgment task [b = −0.88, t(4,460) = −1.178, p > 0.1], it was the opposite for the Time by Response side interaction, which was signiﬁcant in the temporal judgment task [b = 1.93, t(4,460) = 2.592, p < 0.01], but not in the valence judgment task [b = −0.53, t(4,460) = −0.614, p > 0.1]. These separate multilevel analyses revealed that the negative- past/positive-future congruency eﬀect was signiﬁcant in the temporal judgment task [b = −26.27, t(4,465) = −2.330, p < 0.05] but not in the valence judgment task [b = 13.6, t(4,433) = 1.061, p > 0.1]. Of less interest was the signiﬁcant Type of judgment by Valence interaction [b = 54.88, t(8,980) = −4.321, p < 0.001], showing that, while positive verbs elicited faster responses than negative verbs in the temporal judgment task, the opposite was the case in the valence judgment task. Accuracy Analyses The interaction between Type of judgment and Time was marginally signiﬁcant [b = 23.24, t(8,980) = 1.828, p = 0.067]: the tendency to respond faster to the verbs conjugated to the future than to the past tended to be greater in temporal judgment task compared with the valence judgment task. The main eﬀect of Type of judgment was signiﬁcant [b = −143.31, t(8,980) = −15.959, p < 0.001]: participants were faster on the temporal judgment task compared with the valence judgment task. Participants were also faster to respond to negative compared with positive verbs [b = −33.75, t(70) = −2.304, p < 0.05]. Of lesser interest to the purpose of this study, the interaction between Type of judgment and Valence factors was signiﬁcant [b = 3.16, t(9,014) = 3.888, p < 0.001], showing that, contrary to the temporal judgment task, participants were making more errors on negative compared to positive verbs in the valence judgment task. The signiﬁcant interaction between Type of judgment and Time factors [b = −1.67, t(9,014) = −2.052, p < 0.05] revealed that the greater precision of the participants for past compared to future words was stronger in the valence judgment task than in the temporal judgment task. The main eﬀect of Valence [b = −2.15, t(102) = −2.664, p < 0.01] was also signiﬁcant: participants were more precise when judging positive compared to negative words. Discussion In the reaction time analysis, the three-way interaction between Type of judgment, Time and Valence was signiﬁcant [b = −39.36, t(8,980) = −2.191, p < 0.05; see Figure 4]. A separate multilevel model was conducted on each level of the Type of judgment factor that included the two-way interactions of Time by Valence. In Experiment 1, a past-negative/future-positive congruency eﬀect was observed, meaning that a mental metaphor can occur between two concepts with a high level of abstractness. This ﬁnding goes against the previous belief that the mental metaphors June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 8 Spatola et al. The Sad Past Is Left FIGURE 3 \| Percentage of correct trials per condition in Experiment 1, as a function of Type of judgment, Time, and Response side. p < 0.05, *p < 0.01. FIGURE 3 \| Percentage of correct trials per condition in Experiment 1, as a function of Type of judgment, Time, and Response side. p < 0.05, *p < 0.01. saliency processed in secondary task will be processed deeper than a concept with a low level of intrinsic saliency and will more likely be included in a mental model to form a mental metaphor. Accordingly, Santiago et al. (2012) demonstrated that the intrinsic features that contribute to the salience of a conceptual dimension are able to modulate the orientation of exogenous attention. In their study (Experiments 1 and 2), the conceptual dimension of space was processed in the secondary task. The congruency eﬀect between the conceptual domains of valence (processed in the primary task) and space was observed when the salience of the spatial dimension was high (Experiment 1), but not when it was low (Experiment 2). The salience of the spatial dimension was enhanced due to the use of exogenous cues. are necessarily used to map the less complete structures of abstract concepts onto the more complete structures of concrete concepts (Lakoﬀand Johnson, 1980; Boroditsky, 2000). As for the activation of this mental metaphor in the temporal judgment task only, a possibility is that it might be easier to activate this mental metaphor when attention is focused on the dimension of time compared with the dimension of valence. 4Rouget, B. W. (n.d.). Site Francophone sur la Pleine Conscience (Mindfulness) en Psychothérapie. Available online at: http://sites.uclouvain.be/mindfulness/mp3/ BW4.mp3. (Accessed October 01, 2013). 3This mindfulness induction was realized in order to test whether or not the biases associated with the mood of the participant can be modulated, but this topic will be analyzed in a further study that will include the results of a greater number of participants in order to run correlation analyses. Material, Procedure, and Design Experiment 2 replicated Experiment 1 in all aspects, with the following exceptions: each participant carried out the complete experimental set described in Experiment 1 twice, once with a mindfulness induction at the beginning of the experimental session and another without any mindfulness induction. Half the participants received the mindfulness induction in the ﬁrst session and the other half in the second session. One week separated each session. Despite being interesting, it is important to note that most of the eﬀects were observed in the accuracy results only. It was thus essential to replicate the present results, which was the main goal of Experiment 2. EXPERIMENT 2 The mindfulness induction was an audio ﬁle4 presented to the participant via headphones. The transcript and the translation of the audio ﬁle are in Appendix III. Experiment 2 was almost identical to Experiment 1, with the exception that it was repeated twice and in one of the two sessions a mindfulness-based therapy induction was provided to the participants3. Participants h In the valence judgment task only, a left-negative/right- positive congruency eﬀect was observed. This ﬁnding, combined with the fact that the past-negative/future- positive and left-past/right-future congruency eﬀects were observed in the temporal task only, demonstrate that the activation of the mental metaphors is context speciﬁc and conﬁrm that the orientation of attention is important to activate a mental metaphor (Santiago et al., 2011, 2012). The participants were 35 undergraduate students (26 women; mean age = 20.63 years, SD = 2.88) from the same population as in Experiment 1. None of them had participated in the ﬁrst experiment. They all gave informed written consent according to the Declaration of Helsinki. Discussion This phenomenon would result from the daily use of this mental metaphor; people speak of the past and the future in negative and positive terms, but they do not speak of the valence dimension in temporal terms. This repeated use of linguistic expressions would result in the storage of the mental metaphor within the conceptual structure of time (for a similar view, see Casasanto, 2008). Thus, processing the dimension of time but not that of valence would activate this mental metaphor. Another possible explanation is related to the intrinsic saliency of the valence vs. the temporal dimensions. In essence, some studies point out the greater saliency of stimuli charged with emotional content compared to other types of stimuli (e.g., Kuchinke et al., 2005; Nakic et al., 2006). If this were the case here, only the concept of valence would be processed in suﬃcient depth in a secondary task to be able to modulate the processing of other concepts. According to the Coherent Working Models theory (Santiago et al., 2011), a key factor is the intrinsic level of saliency of each dimension. A dimension with a high level of intrinsic In the temporal judgment task, a left-past/right-future congruency eﬀect was also observed, meaning that two mental metaphors can be processed at the same time (see Vicario et al., 2008, for a similar result). Interestingly, since there was no signiﬁcant three-way interaction between the factors of Time, Valence and Response side, it suggests that the activation of these two congruency eﬀects did not depend on the simultaneous use of shared resources. Thus, processing two (or more) mental metaphors that share conceptual compounds would not contribute to form a conglomerate mental metaphor made up of all these conceptual compounds. June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 9 Spatola et al. The Sad Past Is Left FIGURE 4 \| Distribution of RT (in ms) data per condition in Experiment 1, as a function of Type of judgment, Time, and Valence. p < 0.05. FIGURE 4 \| Distribution of RT (in ms) data per condition in Experiment 1, as a function of Type of judgment, Time, and Valence. p < 0.05. Accuracy Analyses In the analysis of reaction times, the Time by Valence interaction was signiﬁcant [b = −24.81, t(18,937) = −4.484, p < 0.001], showing a negative-past/positive-future congruency eﬀect. The Valence by Response side interaction was signiﬁcant [b = −57.02, t(18,938) = −7.256, p < 0.001]. Further, this interaction was modulated by the Type of judgment factor [b = 73.43, t(18,937) = 6.637, p < 0.001] (see Figure 6). Separate multilevel models on each level of the Type of judgment factor showed a signiﬁcant negative-left/positive-right congruency eﬀect in the valence judgment task [b = −56.87, t(9,346) = −6.694, p < 0.001] and, to our surprize, a smaller (8 ms vs. 29 ms) but opposite congruency eﬀect in the temporal task [b = 16.07, t(9,537) = 2.56, p < 0.05). The interaction between Type of judgment, Time and Response side was signiﬁcant, [b = −42.54, t(18,937) = −3.845, p < 0.001] (see Figure 7). Separate multilevel models on each level of the Type of judgment factor revealed a left-past/right- future association signiﬁcant in the temporal judgment task [b = −50.35, t(9,538) = −8.051, p < 0.001] but not in the valence judgment task [b = −9, t(9,371) = −1.059, p > 0.1]. Accuracy analyses revealed that the Time by Valence interaction was signiﬁcant [b = 8, t(19,378) = 12.585, p < 0.001] and this interaction was modulated by the Type of judgment factor [b = −6.23, t(19,378) = −6.932, p < 0.001; see Figure 5]. One separate multilevel model conducted per level of the Type of judgment revealed that the negative-past/positive- future congruency eﬀect was signiﬁcant in both the valence [b = 8, t(9,657) = 11.352, p < 0.001] and the temporal judgment tasks [b = 1.77, t(9,626) = 3.36, p < 0.001], though the congruency eﬀect was greater in the valence (4%) than in temporal judgment tasks (1%). The Valence by Response side interaction was signiﬁcant [b = 1.1, t(19,378) = 2.448, p < 0.05], showing a negative-left/positive-right association. The signiﬁcant Time by Response side interaction b = 2.01, TABLE 3 \| Summary of the results of Experiments 1 and 2 that are of core interest for the main goals of this study. Experiment 1 Experiment 2 Factors ACC RT ACC RT Time × Valence n.s. n.s. * Time × Response side n.s. n.s. * n.s. Valence × Response side n.s. Results the participants3. The data set is deposited in a public repository (Spatola et al., 2017). Three participants were replaced because it was later June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 10 Spatola et al. The Sad Past Is Left t(19,378) = 4.461, p < 0.001] revealed a left-past/right-future congruency eﬀect. discovered that they were non-native French speakers. Responses to the two same verbs as in Experiment 1 (“vaincre,” meaning “win” and “évanouir,” meaning “fainting”) in all conjugated forms were also removed. The accuracy and reaction time results obtained from the remaining participants and items are summarized in Appendix IV. Errors occurred on 4.79% of the trials (1849 trials) and these trials were excluded from the reaction times analyses. Following the same ﬁltering procedure as in Experiment 1, correct trials with a reaction time lower or higher than 2.5 SD per condition for each participant were considered outliers and also excluded from the reaction times analyses, which corresponded to 1,138 trials (3.09% of the trials). The resulting latency and accuracy data were subject to similar analyses as in Experiment 1. The only diﬀerences were that the Session and Mindfulness induction were included as Random eﬀect factors in the models, thereby taking into account the variability that they might induce. Only the signiﬁcant or marginally signiﬁcant ﬁndings (with an alpha level < 0.1) corresponding to these models are reported in the text. The results of core interest are summarized in Table 3. discovered that they were non-native French speakers. Responses to the two same verbs as in Experiment 1 (“vaincre,” meaning “win” and “évanouir,” meaning “fainting”) in all conjugated forms were also removed. The accuracy and reaction time results obtained from the remaining participants and items are summarized in Appendix IV. Errors occurred on 4.79% of the trials (1849 trials) and these trials were excluded from the reaction times analyses. Following the same ﬁltering procedure as in Experiment 1, correct trials with a reaction time lower or higher than 2.5 SD per condition for each participant were considered outliers and also excluded from the reaction times analyses, which corresponded to 1,138 trials (3.09% of the trials). Results Of lesser interest for the purpose of this study, the Type of judgment by Time interaction was signiﬁcant b = 3.74, t(19,378) = 5.888, p < 0.001]: participants were more precise on verbs conjugated to the past compared with the future in the temporal task and it was the opposite in the valence judgment task. The Type of judgment by Valence interaction was signiﬁcant [b = 2.22, t(19,378) = 3.494, p < 0.001], showing that participants were more precise when responding to negative compared with positive words in the valence judgment task only. The main eﬀects of Type of judgment [b = 1.62, t(19,378) = 3.614, p < 0.001], Time [b = −5.24, t(19,378) = −10.425, p < 0.001], Valence [b = −3.57, t(107) = 3.614, p < 0.001] and Response side [b = −1.93, t(19,378) = −4.953, p < 0.001] were signiﬁcant: participants were more precise when judging the verbs’ temporal reference, when responding to verbs conjugated to the past, when these verbs were negative or when they responded with their right hand compared with the opposite. The resulting latency and accuracy data were subject to similar analyses as in Experiment 1. The only diﬀerences were that the Session and Mindfulness induction were included as Random eﬀect factors in the models, thereby taking into account the variability that they might induce. Only the signiﬁcant or marginally signiﬁcant ﬁndings (with an alpha level < 0.1) corresponding to these models are reported in the text. The results of core interest are summarized in Table 3. Accuracy Analyses * *** Type of judgment × Time × Valence n.s. * *** n.s. Type of judgment × Time × Response side * n.s. n.s. * Type of judgment × Valence × Response side n.s. n.s. *** Time × Valence × Response side n.s. n.s. n.s. n.s. Type of judgment × Time × Valence × Response side n.s. n.s. n.s. n.s. p < 0.05, p < 0.01, p < 0.001. ACC is for accuracy and RT is for reaction time results. TABLE 3 \| Summary of the results of Experiments 1 and 2 that are of core interest for the main goals of this study. Of less interest was the signiﬁcant interaction between Type of judgment and Valence [b = −22.01, t(18,939) = −2.816, p < 0.01]: in the valence judgement task, responses tended to be faster for the negative than the positive verbs, whilst there was no observable diﬀerence in the temporal judgment task. There was a signiﬁcant main eﬀect of Type of judgment [b = −164.07, t(18,937) = −24.296, p < 0.001]: participants were faster on the temporal judgment task compared with the valence judgment task. Participants also responded faster to future compared to past verbs [b = 41.83, t(18,941) = 6.743, p < 0.001]. The main eﬀect of Valence was marginally signiﬁcant [b = 24.04, t(66) 1.944, p = 0.056], showing a tendency to respond faster to negative than positive words. The main eﬀect of Response side was also signiﬁcant [b = 41.83, t(18,941) = 6.743, p < 0.001], revealing that the responses on the right were faster than on the left. June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 11 Spatola et al. The Sad Past Is Left FIGURE 5 \| Percentage of correct trials per condition in Experiment 2, as a function of Type of judgment, Time, and Valence. p < 0.001. FIGURE 5 \| Percentage of correct trials per condition in Experiment 2, as a function of Type of judgment, Time, and Valence. *p < 0.001. FIGURE 6 \| Distribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Valence, and Response side. p < 0.05, *p < 0.001. DISCUSSION judgment task in Experiment 2. This congruency eﬀect may be linked to the linear representation of time of the French participants (Yamada and Kato, 2001). According to Guo et al. (2012), the cultures that adopt such a representation of time value the future more positively than the past because it is associated with progress and is something that is still under control, whereas the past is, in some ways, of no value because it cannot be changed. According to Guo et al. (2012), again, this congruency eﬀect should reverse in cultures that adopt a cyclical representation of time because the past helps to predict the future and the future is simply a repetition of the past. Experiment 2 allowed us to conﬁrm several ﬁndings. First, a mental metaphor can occur between two concepts with a high level of abstractness. Second, multiple mental metaphors can be processed at the same time. Third, these simultaneously activated metaphors do not get involved in higher order interactions, and, ﬁnally, the orientation of attention is important to activate a mental metaphor. p The observed congruency eﬀects in Experiment 2 were, in general, greater than those observed in Experiment 1 (see Appendix V for a combined analysis of Experiments 1 and 2). We hypothesized that the repetition of the same experiment, which doubled the number of observations, might have reduced experimental noise. Another possibility is linked to the mindfulness induction, which is known to enhance endogenous attention (Ryan and Deci, 2006; Jha et al., 2007; Moore and Malinowski, 2009). If the participant had more attentional resources to process each conceptual dimension and attention plays a key role in the activation of the mental metaphors (Santiago et al., 2011), then the congruency eﬀects should be stronger. However, a further analysis including Mindfulness as a ﬁxed factor did not allow us to observe a pattern in support of this last possibility. Further research will allow us to determine the factors that contribute to a reduction of the experimental noise when processing mental metaphors. This result adds to previous ﬁndings, showing that mental metaphors, apart from occurring between an abstract and a concrete concept or between two concrete concepts (Rusconi et al., 2006), can also exist between two abstract concepts. GENERAL DISCUSSION The main ﬁndings of the present study can be summarized as follows: (1) a past-negative/future-positive congruency eﬀect was observed only in the temporal judgment task in Experiment 1 and in both tasks in Experiment 2; (2) a left-negative/right- positive congruency eﬀect was observed and this eﬀect was greater in the valence than in the temporal judgment task; (3) a left-past/right-future congruency eﬀect was observed and this eﬀect was greater in the temporal than in the valence judgment task; (4) the three-way interaction between the factors of Time, Valence, and Response side was not signiﬁcant. A further possible account of these types of mental metaphors emerges from the ATOM theory (Walsh, 2003). Because time and valence can be spoken of in terms of more or less (e.g., “I don’t have much time” or “I think that Jim is a better person than Jack”), the magnitude domain of both concepts would be processed by the use of the same neural substrates. This shared use of the “magnitude processor” would result in the congruency eﬀects between both concepts. However, in the task used here, participants were asked to make binary choices (“past”/“future” or “negative”/”positive”), not to judge how much a stimulus was positive or how far in time was the temporal reference of another stimulus. In other words, the domain of magnitude was irrelevant. Further, if that interaction depended on the shared use of a “magnitude processor” and the two other metaphors also depended on the same system, again, a three-way interaction between the factors of Time, Valence, and Response side should have emerged. As a consequence, we consider the Coherent Working Models (Santiago et al., DISCUSSION This argues against the assumption made by Lakoﬀand Johnson (1980), in which the structural borrowing process should necessarily occur from the more complete conceptual structures of the concrete domain toward the less complete conceptual structures of the abstract domain. It is important to note that this congruency eﬀect cannot be attributed to a side eﬀect of the common spatial representations of valence and time. If this were the case, a three-way interaction between the factors of Time, Valence and Response side should have emerged. The Coherent Working Models (Santiago et al., 2011; see also Marmolejo-Ramos et al., in press) appears to be a better candidate to explain the mental metaphors between abstract concepts. Eﬀectively, this theory claims that, when processing a concept, a mental model is built. This mental model would include information from other concepts in order to form a representation that is more complete for the task at hand. The information included in the model should be coherent with the task that has to be carried out with the concept, and the ﬁnal result would be a representation that is more eﬀective for that task. According to this view, the degree to which a concept is abstract or concrete would be irrelevant. Contrary to Experiment 1, the interaction between the concepts of Time and Valence was observed in both tasks and even greater in the valence judgment task than in the temporal judgment task. The possible reasons for such a discrepancy between Experiment 1 and Experiment 2 are taken up in the General Discussion. Accuracy Analyses F ti i P h l \| f ti i 12 J 2018 \| V l 9 \| A ti l 1019 FIGURE 5 \| Percentage of correct trials per condition in Experiment 2, as a function of Type of judgment, Time, and Valence. p < 0.001. FIGURE 6 \| Distribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Valence, and Response side. p < 0.05, **p < 0.001. FIGURE 6 \| Distribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Valence, and Response side. p < 0.05, **p < 0.001. stribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Valence, and Response side. p < 0.0 June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 12 The Sad Past Is Left Spatola et al. Is the Focus of Attention Important for Activating a Mental Metaphor? g p Regarding the third question about the importance of the orientation of attention to activate a mental metaphor, it was possible to observe that the activation of the mental metaphors was task dependent; the left-negative/right-positive congruency eﬀect was greater in the valence than in temporal judgment task, the left-past/right-future congruency eﬀect was greater in the temporal than in valence judgment task, and a signiﬁcant past-negative/future-positive congruency eﬀect was observed in the temporal judgment task in Experiments 1 and 2, and in the valence judgment task in Experiment 2. The fact that the left-past/right-future and left-negative/right-positive congruency eﬀects were manifest mainly in the temporal and valence judgment tasks, respectively, is an indication that attentional cueing plays a key role in the activation of the mental metaphors. In this case, participants were given cues by the type of judgment (on valence or time) and the type of response (manual responses with the left or right hand). This ﬁnding is congruent with those of de la Vega et al. (2012), Santiago et al. (2012) and Torralbo et al. (2006), who showed that the type of judgment and response could serve as cues for the activation of particular mental metaphors. Interestingly, given that in the opposite task some of the congruency eﬀects could be observed, albeit to a lesser extent, it appears that the eﬀects of attentional orienting on the activation of metaphors is a gradual process. Thus the more attention allocated to the concepts of a mental metaphor, the stronger the activation of this metaphor would be. This ﬁnding is rather surprising and appears to be more compatible with the Conceptual Metaphor Theory view (Lakoﬀ and Johnson, 1980, 1999; Boroditsky, 2000). However, recent ﬁndings strongly suggest that the mental metaphors are processed online, in working memory (see Santiago et al., 2011, for a review). We speculate that when participants form and use mental metaphors, they ﬁrst keep active in working memory all the poles of each conceptual dimension in order to form the more coherent representations for the task. Then, when these coherent representations are formed, they are processed as a whole in working memory, without the need to keep active their respective poles. Processing the mental metaphors as a whole would reduce cognitive load and would also be in line with attentional models such as that of Corbetta and Shulman (2002). Can Mental Metaphors Occur Between Two Abstract Concepts? In response to the question “Can mental metaphors occur between two abstract concepts?” the present results indicate that the response is positive. In line with previous results obtained by Guo et al. (2012), a past-negative/future-positive congruency eﬀect was observed in the temporal judgment task in Experiment 1 and in both the temporal and the valence June 2018 \| Volume 9 \| Article 1019 June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 13 Spatola et al. The Sad Past Is Left 2011; see also Marmolejo-Ramos et al., in press) to be the best candidate to explain the metaphorical mapping between two abstract concepts. both cases, the relevant information for the task is selected and organized to form a coherent representation for carrying out the task. These representations, when kept active as a whole, would produce congruency eﬀects such as those observed here. A side eﬀect would be that processing one mental metaphor would not modulate the processing of another mental metaphor because the mapping information would be available in advance and not formed online. More research aimed at testing the time course of the type of information available in working memory while processing mental metaphors is needed to test this hypothesis. Activation? With regard to the second goal of the present study, the present data suggest that two mental metaphors can be processed at the same time, as in Vicario et al’s (2008) study. In Experiment 1, a left-past/right-future and a past-negative/future- positive congruency eﬀects were observed in the temporal judgment task. In Experiment 2, accuracy analyses revealed that the left-past/right-future, left-negative/right-positive and past- negative/future-positive congruency eﬀects were all activated in both the temporal and valence judgment task. Interestingly, the design of the analyses used in this study allowed us to observe that the activations of these mental metaphors were independent from each other. Indeed, the three-way interaction between the factors of Time, Valence, and Response side was not signiﬁcant. Consequently, this means that the distinct congruency eﬀects observed here did not depend on the simultaneous use of shared resources. If the mental metaphors were formed in working memory during the execution of the task (Santiago et al., 2011), each pole of each concept would have been activated in working memory and the information of each pole would have been used to simultaneously form more than one mental metaphor. This competition for the simultaneous use of information corresponding to each conceptual pole would have resulted in a modulation of each of the other mental metaphor processes. The same would have occurred if the mental metaphors were due to the use of shared neural substrates (Walsh, 2003). The multiple mental metaphors would have competed for the use of the shared neural substrates, resulting in a modulation of each of the other mental metaphor processes. Frontiers in Psychology \| www.frontiersin.org Is the Focus of Attention Important for Activating a Mental Metaphor? Corbetta and Shulman (2002) hypothesized that the attentional sets that are formed while performing an attentional task are maintained in working memory during the carrying out of that task. Attentional sets and mental metaphors have much in common in that, in Surprisingly, the past-negative/future-positive congruency eﬀect was signiﬁcant only in the temporal judgment task in Experiment 1 and greater in the valence judgment task in Experiment 2. Here it is important to note that the instructions before starting an experimental block were exactly the same in both experiments and that the irrelevant conceptual dimension (time and valence in the valence and temporal judgment tasks, respectively) was equally cued by the instructions in both judgment tasks (in the temporal task instructions, the words “positive” and “negative” of the valence judgment task were simply interchanged with the words “future” and “past,” respectively). We can think of two possible reasons, one linked to the relative salience of the concepts of time and valence, and the other linked to the activation of diﬀerent mental metaphors. According to the relative salience hypothesis, the past- negative/future-positive congruency eﬀect would have been manifest only in the temporal judgment task in Experiment June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org June 2018 \| Volume 9 \| Article 1019 14 Spatola et al. The Sad Past Is Left FIGURE 7 \| Distribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Time, and Response side. *p < 0.001. FIGURE 7 \| Distribution of RT (in ms) data per condition in Experiment 2, as a function of Type of judgment, Time, and Response side. p < 0.001. ms) data per condition in Experiment 2, as a function of Type of judgment, Time, and Response side. **p < 0.001. 1 because the conceptual dimension of valence is intrinsically more salient than that of time. Some studies suggest that the dimension of valence is eﬀectively more salient than other more neutral dimensions due to connections with emotional neural structures (e.g., Kuchinke et al., 2005; Nakic et al., 2006). As a consequence, the participants would process the concept of valence more deeply in the temporal judgment task than the concept of time in the valence judgment task. Is the Focus of Attention Important for Activating a Mental Metaphor? One could argue that, if the dimension of valence was so active in the temporal task in Experiment 1, a left-negative/right-positive congruency eﬀect should also have been observed in the temporal judgment task, which was not the case. Nevertheless, Santiago et al. (2011) argued that mental metaphors are activated in order to create a coherent and eﬃcient representation of the borrowing concept in relation to the task that has to be carried out with that concept. Since the spatial representation of valence is completely unrelated to the temporal judgment task, this mental metaphor would be of no use and not activated in this task. In Experiment 2, the strong temporal component of the mindfulness induction, by asking the participant to focus on the present moment while performing relaxation exercises (see Appendix II) might have driven participants’ attention to the temporal dimension, making it more salient. As a result, the concept of time in the valence judgment task would be processed with suﬃcient depth to modulate the processing of the valence concept. the valence dimension in their daily life, they would come to store this mental metaphor in long term memory along within the structure of time. As a consequence, this would be readily activated when processing the concept of time, but not that of valence (for a similar view, see Casasanto, 2008). As for Experiment 2, the strong temporal component of the mindfulness induction used in a therapy context might have contributed to the temporalization of the dimension of valence, which also led to a past-negative/future-positive congruency eﬀect in the valence task. The present results do not allow us to distinguish between these two possibilities and these explanations cannot account for the fact that, when Mindfulness was included as a ﬁxed factor in a further analysis, it did not modulate the activation of Time is Valence metaphor in any of the experimental conditions. Further research is needed to directly address this issue. Nevertheless, it is important to note that, no matter what the explanation, these discrepancies between tasks and Experiments highlight the ﬂexibility in the activation of the mental metaphors and this ﬂexibility is predicted by the Coherent Working Models (Santiago et al., 2011). Frontiers in Psychology \| www.frontiersin.org REFERENCES Casasanto, D. (2013). “Development of metaphorical thinking: the role of language,” in Language and the Creative Mind, eds M. Borkent, B. Dancygier, and J. Hinnell (Stanford, CA: CSLI Publications), 3–18. Baayen, R. H., Davidson, D. J., and Bates, M. D. (2008). 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However the LAPSCO lab at University Blaise Pascal had already set up a protocol that adhered to the general principles of the Declaration of Helsinki and we followed it: all participants gave written informed consent, a special care was taken to protect the privacy of the participants and the conﬁdentiality of their personal information, all participants were informed on how to get an appointment with CONCLUSION The present results allowed us to reveal that it is possible to create a mental metaphor between two abstract concepts, and that the focus of attention is important for activating a speciﬁc mental metaphor. Accordingly, the activation of the mental metaphor Another possibility is that the participants of Experiment 1 might have activated a mental metaphor in which the temporal dimension is valued, but not the opposite. Because people frequently value the past and the future but do not temporalize June 2018 \| Volume 9 \| Article 1019 15 Spatola et al. The Sad Past Is Left between the two abstract concepts (time and valence) appeared to depend on attentional factors. These ﬁndings are in line with the Coherent Working Models theory (Santiago et al., 2011), but other results are puzzling. In particular, within the state-of- the-art models already published in the scientiﬁc literature, it is diﬃcult to explain the independent simultaneous activations of diﬀerent mental metaphors together with the need for attention to activate a mental metaphor. Here we proposed an adjustment of the Coherent Working Models theory (Santiago et al., 2011), but more research to directly address this issue will be needed before we endorse this possibility. Moreover, we have yet to determine the precise factors explaining why the congruency eﬀect between the dimensions of valence and time was signiﬁcant only in the temporal judgment task in Experiment 1 whereas it was greater in the valence judgment task in Experiment 2, with a mindfulness induction. It is also unclear why, under certain circumstances, some congruency eﬀects were observed in the accuracy results, but not in the reaction time data (e.g., most of the congruency eﬀects observed in Experiment 1 were only evident in the accuracy data). Future research will therefore be necessary to shed light on the characteristics of the mechanisms responsible for these eﬀects. the counseling and psychological services at that same university if they thought they needed it after responding to the HAD questionnaire, and an external person was in charge to receive and process any possible complaint. FUNDING The present research was carried out as partial fulﬁllment of the requirements for NS’s Ph.D. thesis. NS’s was supported by a Ph.D. research fellowship from the École doctorale LSHS, University Blaise Pascal, Clermont-Ferrand, France. MO was supported by a Juan de la Cierva fellowship (JCI-2012-13046) from the Spanish Ministry of Education and Science. Research was funded by research projects PSI2012-32464 (Spanish Ministry of Economy and Competitiveness) and PSI2015-67531-P (Spanish Ministry of Economy and Competitiveness) to JS. AUTHOR CONTRIBUTIONS MO led the study. His involvement as that of NS included designing the study, collecting the data, analyzing the data, and writing the manuscript. BB made a signiﬁcant contribution to the design of the study, he was involved in data collection, and also contributed to data interpretation and manuscript writing. MM, LF and JS made signiﬁcant contributions to the design of the study, the data analysis and interpretation, and to the writing of the manuscript. REFERENCES Time orientation in the United State, China and Mexico, measurement and insights for promotional strategy. J. Int. Consum. Market. 13, 57–75. doi: 10.1300/J046v13n01_05 g p Consum. Market. 13, 57–75. doi: 10.1300/J046v13n01_05 Marmolejo-Ramos, F., Khatin-Zadeh, O., Yazdani-Fazlabadi, B., Tirado, C., and Sagi, E. 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Sci. 19, 838–842. doi: 10.1111/j.1467- 9280.2008.02165.x Ouellet, M., Santiago, J., Funes, M. J., and Lupiáñez, J. (2010a). Thinking about the future moves attention to the right. J. Exp. Psychol. Hum. Percept. Perform. 36, 17–24. doi: 10.1037/a0017176 Ouellet, M., Santiago, J., Israeli, Z., and Gabay, S. (2010b). Is the future the right time? Exp. Psychol. 57, 308–314. doi: 10.1027/1618-3169/a0 00036 Zhong, C. B., and Liljenquist, K. (2006). Wash away your sins: threatened morality and physical cleansing. Science 313, 1451–1452. doi: 10.1126/science.1130726 Razavi, D., Delvaux, N., Farvacques, C., and Robaye, E. (1989). REFERENCES 38, 1030–1040. doi: 10.1177/0146167212443895 June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 16 Spatola et al. The Sad Past Is Left Jha, A. P., Krompinger, J., and Baime, M. J. (2007). Mindfulness training modiﬁes subsystems of attention. Cogn. Aﬀect. Behav. 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Copyright © 2018 Spatola, Santiago, Beﬀara, Mermillod, Ferrand and Ouellet. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. R Core Team (2017). R: A Language and Environment for Statistical Computing. Vienna: R Foundation for Statistical Computing. Available online at: https:// www.R-project.org/ Roitman, J. D., Brannon, E. M., Andrews, J. R., and Platt, M. L. (2007). Nonverbal representation of time and number in adults. Acta Psychol. 124, 296–318. doi: 10.1016/j.actpsy.2006.03.008 June 2018 \| Volume 9 \| Article 1019 Frontiers in Psychology \| www.frontiersin.org 17
https://openalex.org/W4318416222	https://peerj.com/articles/14716v0.3/submission	English	null	Peer Review #3 of "RNAi-based knockdown of candidate gut receptor genes altered the susceptibility of Spodoptera frugiperda and S. litura larvae to a chimeric toxin Cry1AcF (v0.2)"	null	2,023	cc-by	13,123	Manuscript to be reviewed RNAi-based knockdown of candidate gut receptor genes altered the susceptibility of Spodoptera frugiperda and S. litura larvae to a chimeric toxin Cry1AcF p y p p g p litura larvae to a chimeric toxin Cry1AcF Tushar K. Dutta Corresp., 1 , Kodhandaraman Santhoshkumar 1 , Arudhimath Veeresh 1 , Chandramani Waghma Chetna Mathur 1 , Rohini Sreevathsa 2 1 Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, Delhi, India 2 ICAR-National Institute for Plant Biotechnology, New Delhi, Delhi, India Corresponding Author: Tushar K. Dutta Email address: tushar.dutta@icar.gov.in Tushar K. Dutta Corresp., 1 , Kodhandaraman Santhoshkumar 1 , Arudhimath Veeresh 1 , Chandramani Waghmare 1 , Chetna Mathur 1 , Rohini Sreevathsa 2 1 Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, Delhi, India 2 ICAR-National Institute for Plant Biotechnology, New Delhi, Delhi, India Corresponding Author: Tushar K. Dutta Email address: tushar.dutta@icar.gov.in 1 Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, Delhi, India 2 ICAR-National Institute for Plant Biotechnology, New Delhi, Delhi, India Background: A multitude of Cry toxins (secreted by Bacillus thuringiensis or Bt) has been deployed globally either via transgenic mean or bio-pesticidal formulations in order to manage insect pests. However, Bt resistance development in insects is emerging as a major concern. To avoid this problem, multiple gene pyramiding or protein-engineered chimeric toxin-based strategy has been analyzed. Methods: In the present study, one such chimeric toxin Cry1AcF (contain the swapped domains of Cry1Ac and Cry1F) was used to investigate its in vivo pathogenesis process in lepidopteran pests Spodoptera frugiperda and S. litura. A number of biochemical and molecular analysis were performed. Results: Oral ingestion of Cry1AcF caused greater toxicity in S. frugiperda than S. litura with larvae displaying increased hemolymph melanization. Histopathology of the midgut transverse sections exhibited Cry1AcF-induced extensive gut damage in both the test insects followed by cytotoxicity in terms of reduced hemocyte numbers and viability. Elevated hemolymph phenoloxidase activity indicated the immune-stimulatory nature of Cry1AcF. In order to analyze the role of gut receptor proteins in Cry1AcF intoxication in test insects, we performed RNAi-mediated silencing using bacterially-expressed dsRNAs of individual receptor-encoding genes including CAD, ABCC2, ALP1 and APN. Target-speciﬁc induced downregulation of receptor mRNAs diﬀerentially altered the insect susceptibility to Cry1AcF toxin in our study. The susceptibility of ALP1 and APN dsRNA pre-treated S. frugiperda was considerably decreased when treated with Cry1AcF in LD50 and LD90 doses. Whereas, susceptibility of CAD and ABCC2 dsRNA pre-treated S. litura was signiﬁcantly reduced when ingested with Cry1AcF in diﬀerent doses. CAD/ABCC2-silenced S. frugiperda and ALP1/APN-silenced S. litura were vulnerable to Cry1AcF alike of control larvae. In conclusion, our results indicate ALP1/APN and CAD/ABCC2 as the functional receptor for Cry1AcF toxicity in S. frugiperda and S. litura, respectively. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Manuscript to be reviewed 1 2 RNAi-based knockdown of candidate gut receptor genes altere 3 susceptibility of Spodoptera frugiperda and S. litura larvae to a 4 Cry1AcF 5 Tushar K. Tushar K. Dutta Corresp., 1 , Kodhandaraman Santhoshkumar 1 , Arudhimath Veeresh 1 , Chandramani Waghmare 1 , Chetna Mathur 1 , Rohini Sreevathsa 2 Dutta1, Kodhandaraman Santhoshkumar1, Arudhimath Veer 6 Chandramani Waghmare1, Chetna Mathur1, Rohini Sreevathsa2 7 1Division of Nematology, ICAR-Indian Agricultural Research Institute, New 8 India 9 2ICAR-National Institute for Plant Biotechnology, New Delhi, 110012, India 10 11 12 Corresponding Author 13 Dr. Tushar K Dutta 14 Division of Nematology 15 ICAR-Indian Agricultural Research Institute 16 New Delhi, India-110012 17 TEL: +91-11-2584-2721 18 Email: tushar.dutta@icar.gov.in; nemaiari@gmail.com 19 20 21 22 23 24 25 26 27 28 PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 29 Abstract 30 Background: A multitude of Cry toxins (secreted by Bacillus thuringiensis or Bt) has been 31 deployed globally either via transgenic mean or bio-pesticidal formulations in order to manage 32 insect pests. However, Bt resistance development in insects is emerging as a major concern. To 33 avoid this problem, multiple gene pyramiding or protein-engineered chimeric toxin-based 34 strategy has been analyzed. 30 Background: A multitude of Cry toxins (secreted by Bacillus thuringiensis or Bt) has been 31 deployed globally either via transgenic mean or bio-pesticidal formulations in order to manage 32 insect pests. However, Bt resistance development in insects is emerging as a major concern. To 33 avoid this problem, multiple gene pyramiding or protein-engineered chimeric toxin-based 34 strategy has been analyzed. 30 Background: A multitude of Cry toxins (secreted by Bacillus thuringiensis or Bt) has been 31 deployed globally either via transgenic mean or bio-pesticidal formulations in order to manage 32 insect pests. However, Bt resistance development in insects is emerging as a major concern. To 33 avoid this problem, multiple gene pyramiding or protein-engineered chimeric toxin-based 34 strategy has been analyzed. 35 Methods: In the present study, one such chimeric toxin Cry1AcF (contain the swapped domains 36 of Cry1Ac and Cry1F) was used to investigate its in vivo pathogenesis process in lepidopteran 37 pests Spodoptera frugiperda and S. litura. A number of biochemical and molecular analysis were 38 performed. 35 Methods: In the present study, one such chimeric toxin Cry1AcF (contain the swapped domains 36 of Cry1Ac and Cry1F) was used to investigate its in vivo pathogenesis process in lepidopteran 37 pests Spodoptera frugiperda and S. litura. A number of biochemical and molecular analysis were 38 performed. 35 Methods: In the present study, one such chimeric toxin Cry1AcF (contain the swapped domains 36 of Cry1Ac and Cry1F) was used to investigate its in vivo pathogenesis process in lepidopteran 37 pests Spodoptera frugiperda and S. litura. A number of biochemical and molecular analysis were 38 performed. 39 Results: Oral ingestion of Cry1AcF caused greater toxicity in S. frugiperda than S. litura with 40 larvae displaying increased hemolymph melanization. Histopathology of the midgut transverse 41 sections exhibited Cry1AcF-induced extensive gut damage in both the test insects followed by 42 cytotoxicity in terms of reduced hemocyte numbers and viability. Elevated hemolymph 43 phenoloxidase activity indicated the immune-stimulatory nature of Cry1AcF. 1 2 RNAi-based knockdown of candidate gut receptor genes altered the 3 susceptibility of Spodoptera frugiperda and S. litura larvae to a chimeric toxin 4 Cry1AcF 5 Tushar K. Dutta1, Kodhandaraman Santhoshkumar1, Arudhimath Veeresh1, 6 Chandramani Waghmare1, Chetna Mathur1, Rohini Sreevathsa2 7 1Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, 8 India 9 2ICAR-National Institute for Plant Biotechnology, New Delhi, 110012, India 10 11 12 Corresponding Author 13 Dr. Tushar K Dutta 14 Division of Nematology 15 ICAR-Indian Agricultural Research Institute 16 New Delhi, India-110012 17 TEL: +91-11-2584-2721 18 Email: tushar.dutta@icar.gov.in; nemaiari@gmail.com 19 20 21 22 23 24 25 26 27 28 1 28 PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Manuscript to be reviewed 58 Introduction 59 According to a United Nations report, the current global human population (7.9 billion) may 60 grow to 8.5 and 9.7 billion in 2030 and 2050, respectively 61 (https://www.un.org/en/observances/world-population-day). This ever-growing population has 62 put an asymmetric demand on food supply especially when global agriculture is facing shrinkage 63 of cultivable land, water crisis and arrival of new pests and pathogens due to climate change 64 (IPPC Secretariat 2021). Although protected agriculture and vertical farming are rapidly being 65 adopted, insect pests are emerging as a major problem due to the conducive environment 66 maintained inside the protected structures (Phani et al. 2021). Indian agriculture suffers heavily 67 due to a multitude of insect pest occurrence including noctuid lepidopteran pests such as 68 Spodoptera litura and S. frugiperda (Dhaliwal et al. 2015; EFSA Panel on Plant Health 2019; 69 Suby et al. 2020). The polyphagous S. litura can infect over 100 crops and are widely distributed 70 across tropical and subtropical Asia including India, China and Japan (Cheng et al. 2017). In 71 Africa, America and Asia (including Indian subcontinent), the fall armyworm (S. frugiperda) has 72 become an invasive species (mostly in cotton and cultivated grasses such as maize, sorghum, 73 rice, sugarcane etc.) because of the global warming effects (CABI Compendium 2019; IPPC 74 Secretariat 2021). 59 According to a United Nations report, the current global human population (7.9 billion) may 60 grow to 8.5 and 9.7 billion in 2030 and 2050, respectively 59 According to a United Nations report, the current global human population (7.9 billion) may 60 grow to 8.5 and 9.7 billion in 2030 and 2050, respectively 75 Synthetic pesticides are continually being withdrawn from market or placed under restricted use 76 because of the insecticide resistance emergence in insects, lesser degradability of pesticides in 77 soil, avian and mammalian toxicity of insecticide residues etc. As a result, very few active 78 insecticides are at disposal for the growers (http://ppqs.gov.in/). Use of biological control agents 79 such as Bacillus thuringiensis (Bt) for pest control either via spraying or transgenic mean has 80 been met with great success (Koch et al. 2015; Liu et al. 2021). Bt secretes an insecticidal 81 crystalline protein popularly known as delta endotoxin or Cry toxin which kills the insect (Bravo 82 et al. 2007). 29 Abstract In order to analyze 44 the role of gut receptor proteins in Cry1AcF intoxication in test insects, we performed RNAi- 45 mediated silencing using bacterially-expressed dsRNAs of individual receptor-encoding genes 46 including CAD, ABCC2, ALP1 and APN. Target-specific induced downregulation of receptor 47 mRNAs differentially altered the insect susceptibility to Cry1AcF toxin in our study. The 48 susceptibility of ALP1 and APN dsRNA pre-treated S. frugiperda was considerably decreased 49 when treated with Cry1AcF in LD50 and LD90 doses. Whereas, susceptibility of CAD and 50 ABCC2 dsRNA pre-treated S. litura was significantly reduced when ingested with Cry1AcF in 51 different doses. CAD/ABCC2-silenced S. frugiperda and ALP1/APN-silenced S. litura were 52 vulnerable to Cry1AcF alike of control larvae. In conclusion, our results indicate ALP1/APN and 53 CAD/ABCC2 as the functional receptor for Cry1AcF toxicity in S. frugiperda and S. litura, 54 respectively. 57 PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 88 transporters located in the epithelial cell membrane of the insect midgut. Binding of Cry protein 89 with these receptors lead to oligomerization of Cry monomers that ultimately lead to cytotoxicity 90 (Zhang et al. 2006; Adang et al. 2014; Jurat-Fuentes et al. 2021). Domain I of Cry toxin aids in 91 tethering to the cell membrane via its hydrophobic helical hairpin, Domain II helps in detecting 92 putative receptors via its highly variable loops and Domain III is associated with receptor 93 binding and catalytic activity (Chakrabarty et al. 2022). 94 To date, a repertoire of Cry toxins has been characterized globally for their future deployment 95 via transgenic mean. The sustained use of this technology was demonstrated when a number of 96 those toxin genes were expressed in Bt crops (Carrière et al. 2019; Dutta et al. 2022c). g p p ( ; ) 97 Nevertheless, reports of resistance development in insects towards different Cry protein encoding 98 genes owing to wider deployment of narrow-spectrum Bt crops in India, China, Brazil, Argentina 99 and United States have become an alarming concern (Tabashnik et al. 2013; Tabashnik and 100 Carrière 2017; Carrière et al. 2019). In order to minimize the Bt resistance development in 101 insects, multiple gene pyramiding or protein-engineered chimeric toxin-based strategy has been 102 advocated (Honée et al. 1990; Ho et al. 2006; Tajne et al. 2014; Zghal et al. 2017; Zafar et al. 103 2020; Chen et al. 2021). It is assumed that chimeric toxins would have broad-spectrum efficacy 104 that would aid in averting Bt resistance development in insects (Rathinam et al. 2019). One such 105 chimeric toxin was generated by fusing Domain I and II of Cry1Ac with Domain III of Cry1F 106 and named as Cry1AcF, which is an activated toxin (patent no 237912; Rathinam et al. 2017). 107 This chimeric toxin when expressed via transgenic mean in different pulse crops it showed 108 resistance against the lepidopteran pest Helicoverpa armigera in confined trials (Keshamma et 109 al. 2012; Ramu et al. 2012; Muralimohan et al. 2020; Ramkumar et al. 2020). However, 110 information about the role of this toxin in insect in vivo pathogenesis was lacking. Manuscript to be reviewed In our recent 111 studies, we showed that Cry1AcF can negatively alter the midgut architecture of the model insect 112 Galleria mellonella and the toxin requires to bind with different gut receptors to become 113 insecticidal (Dutta et al. 2022a,b,c). In the present study, we analyzed the in vivo pathogenesis of 114 Cry1AcF in economically important insects such as S. frugiperda and S. litura, and the putative 115 role of gut receptors in Cry1AcF intoxication process was established. 58 Introduction Worldwide, the acreage of Bt transgenic crops has increased tremendously since 83 their introduction during 1996 (ISAAA 2019). Currently, a number of Bt crops are commercially 84 grown including Bt maize, Bt cotton, Bt soybean, Bt rice and Bt eggplant (Chakrabarty et al. 85 2022). Cry intoxication mechanism may involve pore formation or cell signaling model. In either 86 cases, the activated toxin has to bind with different receptors such as cadherin (CAD), 87 aminopeptidase N (APN), alkaline phosphatase (ALP) and ATP-binding cassette (ABC) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 118 S. frugiperda and S. litura egg clusters were obtained from infested plots (IARI, New Delhi) of 119 maize and okra followed by initiation of nucleus colony culture in our laboratory. Rearing of 120 neonates was performed in sterilized castor leaves in a growth chamber; sixth-instar larvae were 121 pupated in sterilized sand followed by disinfection of pupae via rinsing in 0.02% formaldehyde. 122 Adults were fed with artificial diets comprising honey, multivitamin supplements, ascorbic acid 123 and methyl-p-hydroxybenzoate (Shankhu et al. 2020; Dutta et al. 2021). A continuous culture 124 was maintained on castor leaves for recurrent insect life cycle progression. Surface-sterilized 125 (with 70% ethanol) fourth-instar larvae (0.4-0.5 g body mass) were used for subsequent 126 experiments. 127 Cry1AcF toxicity assessment in test insects 128 Larvae were randomly assigned to each treatment. Larvae were starved for 6 h followed by oral 129 ingestion with 10 µL of phosphate-buffered saline (PBS, pH 7.4) containing Cry1AcF toxin in 130 different doses using a sterilized 26-gauge hypodermic needle (Hamilton syringe, Sigma- 131 Aldrich). Negative control consisted of PBS only. A known toxin TcaB (Mathur et al. 2019; 132 Dutta et al. 2021) was used as the positive control. Larvae were incubated in sterile 6-well 133 polystyrene plates containing artificial diet at 28ºC, 60% relative humidity. Insect mortality data 134 was recorded at 24 h after oral ingestion. The whole experiment was repeated at least thrice (n = 135 50 per treatment). The details of Cry1AcF protein production and purification are described in 136 our previous studies (Rathinam et al. 2017, 2019). 117 Insect rearing PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 148 haematoxylin and eosin (Sigma-Aldrich). Samples were mounted in Dibutylphthalate 149 Polysterene Xylene (DPX, Sigma-Aldrich). More detail about the procedure can be found in 150 Dutta et al. (2021) and Santhoshkumar et al. (2021). Photomicrographs obtained in Zeiss 151 Axiocam MRm microscope. 148 haematoxylin and eosin (Sigma-Aldrich). Samples were mounted in Dibutylphthalate 149 Polysterene Xylene (DPX, Sigma-Aldrich). More detail about the procedure can be found in 150 Dutta et al. (2021) and Santhoshkumar et al. (2021). Photomicrographs obtained in Zeiss 151 Axiocam MRm microscope. 162 Cry1AcF immune-stimulatory activity measurement in test insects 163 Phenoloxidase (PO) enzyme activity was measured in the hemolymph of toxin-treated and 164 control larvae at 12 and 24 h after inoculation. 10 µL hemolymph was extracted from each larvae 165 as described above and 100 µL PBS was added to it by centrifugation at 1006 g for 2 min at 4°C. 166 Cell-free supernatant was used for downstream analyses. 100 µL of hemolymph sample (cell- 167 free supernatant) was added to 100 µL of 20 mM L-3,4-dihydroxyphenylalanine or L-DOPA (4 168 mg mL-1 in PBS) in a 96-well ELISA plate and incubated at 28°C in dark for 30 min (Mathur et 169 al. 2019; Dutta et al. 2021). Absorbance of the sample was examined at 490 nm in a microplate 170 reader (BioTek). PO activity was expressed as the increase in absorbance at 490 nm per minute 171 per mg of protein. As control, PBS and L-DOPA were mixed equally and its absorbance value 172 was subtracted from each sample. Experiment was repeated thrice (n = 10 per treatment). 152 Cry1AcF cytotoxicity analysis in test insects 153 Total hemocyte counts (THC) were obtained from toxin-treated and control larvae at 24 h after 154 oral ingestion. Sample of fresh hemolymph (10 µL) was extracted (by piercing through the 155 integument above head using a sterilized needle) from each larvae and diluted three-fold with 156 ice-cold anticoagulant containing phenylthiourea (4 mg mL-1 in PBS) to suppress the 157 melanization (Mathur et al. 2019). To examine hemocyte viability, 0.4% Trypan blue (w/v in 158 PBS) was mixed to the hemolymph, which was not treated with the anticoagulant (Mathur et al. 159 2019). On both occasions, hemolymph sample was applied into a hemocytometer (Neubauer) for 160 cell enumeration in Zeiss Axiocam MRm microscope. Experiment was repeated thrice (n = 10 161 per treatment). 137 Histopathology study 138 At 24 h after oral ingestion, toxin-treated and control larvae were inoculated with 100 µL of 10% 139 formalin via intra-hemocoel injection followed by dipping the larvae in formalin (Shankhu et al. 140 2020). After 24 h, midgut tissue was dissected out using a sterilized blade under the microscope, 141 and suspended in ice-cold fixative (4% paraformaldehyde + 2.5% glutaraldehyde in 0.1 M PBS, 142 pH 7.2) followed by transfer to fresh fixative and overnight storing at 4°C. Next, samples 143 dehydrated through a graded ethanol series. Dehydrated samples were immersed in xylene for 144 clearing of tissues. Embedded (in paraffin wax) samples were processed into 6 µm cross-sections 145 using Leica RM2165 microtome (Dutta et al. 2021; Santhoshkumar et al. 2021). Sections fixed 146 in glass slides via egg albumin solution and dried overnight. Slides washed using graded ethanol 147 series and sections dewaxed via xylene treatment. Sections were periodically stained with Cole’s PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 182 RT-qPCR analysis 183 RT-qPCR-based transcription profile analysis of Cry receptor-encoding genes was performed in 184 a CFX96 thermal cycler (BioRad). qPCR reaction volume (10 μL) consisted of 1.5 ng cDNA, 185 forward and reverse primers (750 nM each), and 5 μL SYBR Green PCR master-mix (BioRad). 186 qPCR cycling conditions included a hot start phase of 95°C for 30 s, followed by 40 cycles of 187 95°C for 10 s and 60°C for 30 s. Additionally, a melt curve program (95°C for 15 s, 60°C for 15 188 s, followed by a slow ramp from 60 to 95°C) was included to ascertain the amplification 189 specificity. Quantification cycle (Cq) values were obtained from CFX Maestro software 190 (BioRad). Housekeeping genes of S. frugiperda (ribosomal protein S3 or rps3) and S. litura (β- 191 actin and GAPDH) were included as the internal reference for normalizing the expression value 192 of target genes. qPCR fold change data was calculated using 2-ΔΔCq method. qPCR was run with 193 five biological and three technical replicates for each samples. Primer designing was carried out 194 in OligoAnalyzer tool (https://eu.idtdna.com/). PCR reaction efficiency of different primers was 195 determined by constructing a standard curve (where Cq values were plotted against cDNA 196 concentrations) across the five-fold dilutions of cDNA samples followed by calculating the 197 slope using a linear regression equation: E = (10(-1/slope) – 1) × 100. Primer details and PCR 198 efficiency data are given in Supplementary Tables 1 and 2. Manuscript to be reviewed 177 In order to minimize genomic DNA contamination, extracted RNA was digested with DNase I 178 enzyme (TakaRa). Purity of the RNA was assessed in a Nanodrop ND-1000 spectrophotometer 179 (Thermo Fisher Scientific), and RNA integrity was evaluated by electrophoresing samples on 1% 180 (w/v) agarose gel. RNA (~ 1 µg) was reverse transcribed to cDNA by using a first-strand cDNA 181 synthesis kit (Superscript VILO, Invitrogen). 173 RNA extraction from S. frugiperda and S. litura 174 To analyze the transcription dynamics of different Cry receptor-encoding genes in different 175 developmental stages and tissues of S. frugiperda and S. litura, RNA was extracted from 176 different samples using TRIzol reagent (Invitrogen) by following the manufacturer's protocols. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 207 harbored SacI and HindIII restriction endonuclease sites (Supplementary Tables 1 and 2). PCR 208 products were cloned into SacI and HindIII-digested L4440 plasmid (Addgene) that contained 209 two T7 polymerase promoter sequences (aids in driving DNA to RNA transcription) in inverted 210 orientation flanking the multiple cloning site. DsRNA molecules were bacterially expressed in 211 Escherichia coli HT115(DE3) cells, which were transformed with recombinant L4440 plasmid. 212 HT115 cells were cultured in Luria Bertani medium (Sigma) containing ampicillin (50 µg mL-1) 213 and tetracycline (12.5 µg mL-1) at 37°C for 12 h with constant agitation at 200 rpm. In order to 214 induce T7 polymerase synthesis, 0.4 mM IPTG was mixed into the culture and incubated for an 215 additional 4 h at 37°C. Transcribed dsRNAs were isolated from bacterial aliquots (dsRNAs 216 purified from E. coli cells by phenol/chloroform/isoamyl alcohol extraction via shaking at 65ºC 217 for 30 min followed by centrifugation at 12000 × g for 15 min, upper phase containing dsRNA 218 transferred to a new tube, precipitated with isopropanol, washed with ethanol, followed by re- 219 suspending the nucleic acid pellet in 0.05 M PBS) and electrophoresed on a 1% (w/v) agarose 220 gel (Ahn et al. 2019). DsRNA of a gfp gene (Genbank ID: HF675000) ligated into L4440 221 plasmid was used as the non-native control (in order to check whether dsRNA itself has any 222 negative effect on insects). 199 DsRNA synthesis 200 Targeted region for dsRNA synthesis from the coding sequences of different receptors was 201 designed using different in silico tools such as siDirect (http://sidirect2.rnai.jp/; predicts siRNA 202 biogenesis probability across the query sequence), Dharmacon (http://horizondiscovery.com/) 203 and dsCheck (http://dscheck.rnai.jp/; helps in avoiding off-target sites by comparing sequence 204 homology of predicted siRNAs with that from non-target organism). DsRNA sequences were 205 PCR-amplified (according to the standard protocol) from the cDNAs of S. frugiperda and S. 206 litura (extracted from the midgut tissues of fourth-instar larvae) using gene-specific primers that PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 244 Biological activity of Cry1AcF against S. frugiperda and S. litura 245 Oral ingestion of Cry1AcF (Fig 1A) caused toxicity in the fourth-instar larvae of S. frugiperda 246 and S. litura. Phenotypic changes such as dead-like posture with extended prolegs and 247 hemolymph melanization was evident in Cry1AcF-treated larvae compared to normal 248 phenotypes in PBS-ingested control larvae at 24 h after inoculation (Fig 1B). Cry1AcF conferred 249 a gradual and significant (P < 0.01) decrease in mean percent survival of S. frugiperda (F (4, 45) 250 = 56.82, P = 0.0021) and S. litura (F (4, 45) = 47.38, P = 0.0032) with increasing Cry1AcF 251 doses, i.e. 25-200 ng/larva (Fig 1C, D). While compared with a gut-active reference toxin TcaB 252 (characterized from an insect parasitic bacterium Photorhabdus akhurstii in our laboratory; 253 Mathur et al. 2019; Santhoshkumar et al. 2021), Cry1AcF caused significantly (P < 0.01) greater 254 larval mortality across the dose range in both the test insects (Fig 1C, D). Consequently, 255 Cry1AcF exhibited lower LD50 and LD90 values than TcaB in both the test insects (Table 1). The 256 LD50 and LD90 values for Cry1AcF in S. frugiperda was derived as 29.78 and 99.77 ng/larva, 257 respectively. The LD50 and LD90 values for Cry1AcF in S. litura was calculated as 38.76 and 258 117.32 ng/larva, respectively (Table 1). When larval body mass was included in calculation, 259 Cry1AcF caused greater toxicity in S. frugiperda in terms of lower LD50 (53.60 ng/g) and LD90 260 (179.59 ng/g) values compared to S. litura (LD50: 69.77 ng/g; LD90: 211.18 ng/g). Manuscript to be reviewed Manuscript to be reviewed 223 RNAi bioassay 224 Bacterially-expressed dsRNAs (10 µL suspension amounting to ~10 µg of dsRNA) were orally 225 administered into the 6 h starved fourth-instar larvae of S. frugiperda or S. litura using a 226 sterilized needle as described above. GFP dsRNA and 0.05 M PBS served as the non-native and 227 negative control, respectively. Inoculated larvae were incubated in sterile 6-well plates as 228 described above. Post 24 h of dsRNA treatment, larvae were orally ingested with LD50 or LD90 229 doses of Cry1AcF toxin and incubated in 6-well plates as depicted above. After 24 h insect 230 mortality data was recorded. The whole experiment was repeated at least thrice (n = 50 per 231 treatment). 232 RNAi-induced downregulation (fold change in expression of a target gene in dsRNA-treated 233 larvae was subtracted from that in control larvae) of target receptor-encoding genes was 234 validated by RT-qPCR assay. RNA was isolated from the midgut tissues of ten random larvae 235 belonging to each dsRNA treatment groups. RNA was reverse-transcribed to cDNA as depicted 236 previously. qPCR run conditions were followed as explained above. 224 Bacterially-expressed dsRNAs (10 µL suspension amounting to ~10 µg of dsRNA) were orally 225 administered into the 6 h starved fourth-instar larvae of S. frugiperda or S. litura using a 226 sterilized needle as described above. GFP dsRNA and 0.05 M PBS served as the non-native and 227 negative control, respectively. Inoculated larvae were incubated in sterile 6-well plates as 228 described above. Post 24 h of dsRNA treatment, larvae were orally ingested with LD50 or LD90 229 doses of Cry1AcF toxin and incubated in 6-well plates as depicted above. After 24 h insect 230 mortality data was recorded. The whole experiment was repeated at least thrice (n = 50 per 231 treatment). 232 RNAi-induced downregulation (fold change in expression of a target gene in dsRNA-treated 233 larvae was subtracted from that in control larvae) of target receptor-encoding genes was 234 validated by RT-qPCR assay. RNA was isolated from the midgut tissues of ten random larvae 235 belonging to each dsRNA treatment groups. RNA was reverse-transcribed to cDNA as depicted 236 previously. qPCR run conditions were followed as explained above. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 237 Statistical analysis 238 LD50 and LD90 doses of Cry1AcF was determined by using probit analysis in SPSS v. 21 239 software (IBM Corp., NY, USA). Data of different bioassay (THC and PO assay data were 240 checked for normality using Shapiro-Wilk test) and qPCR experiments was subjected to one-way 241 ANOVA followed by Tukey’s honest significant difference (HSD) test (for multiple comparison 242 among different treatments) in SAS v. 14.1 software. 238 LD50 and LD90 doses of Cry1AcF was determined by using probit analysis in SPSS v. 21 239 software (IBM Corp., NY, USA). Data of different bioassay (THC and PO assay data were 240 checked for normality using Shapiro-Wilk test) and qPCR experiments was subjected to one-way 241 ANOVA followed by Tukey’s honest significant difference (HSD) test (for multiple comparison 242 among different treatments) in SAS v. 14.1 software. Manuscript to be reviewed 266 inoculation. Midgut tissues of PBS-ingested larvae exhibited normal morphology in terms of 267 regular arrangement of epithelial cells (with intact nucleus) which were resting on the basement 268 membrane. Gut tissue damage was negligible because the protective barrier provided by visceral 269 muscle layer (interface between gut epithelium and hemocoel or body cavity) was intact in both 270 the test insects (Figure 2A, C). By contrast, in Cry1AcF-ingested larvae, epithelial cell lining 271 was displaced, cells were disintegrated and sloughed off into the gut lumen. Additionally, a 272 discontinuous lining of visceral muscle layer was evident suggesting the structural damage to 273 gut-body cavity barrier in the test insects (Figure 2B, D). 274 Cry1AcF reduces hemocyte numbers and viability in S. frugiperda and S. litura 275 Interestingly, the cytotoxic effect of Cry1AcF was observed in both the test insects at 24 h after 276 oral ingestion. Total circulatory hemocyte counts (THC) of S. frugiperda was significantly (F (2, 277 27) = 24.44, P = 0.0072) reduced from 5.99 × 106 mL-1 in PBS to 3.48 × 106 mL-1 and 2.27 × 106 278 mL-1 in Cry1AcF-treated insects at LD50 (30 ng/larva) and LD90 (100 ng/larva) doses, 279 respectively. Similarly, THC of S. litura was significantly (F (2, 27) = 29.37, P = 0.0058) 280 declined from 9.88 × 106 mL-1 in PBS to 6.89 × 106 mL-1 and 4.36 × 106 mL-1 in Cry1AcF- 281 treated insects at LD50 (40 ng/larva) and LD90 (120 ng/larva) doses, respectively (Figure 3A). 282 In addition, viability of hemocyte cells was examined by Trypan blue staining assay. In this 283 assay, viable cells avert the stain penetration while dead cells dye dark blue because of stain 284 penetration via disintegrated cell membrane. Approximately 100 cells were sampled from each 285 treatment (n = 10), live cells were counted and expressed as percent viability. Cell viability of S. 286 frugiperda was significantly (F (2, 27) = 16.77, P = 0.0091) reduced from 100% in PBS to 65% 287 and 43% in Cry1AcF-treated insects at LD50 and LD90 doses, respectively. Likewise, cell 288 viability of S. litura was significantly (F (2, 27) = 14.88, P = 0.0062) decreased from 100% in 289 PBS to 69% and 49% in Cry1AcF-treated insects at LD50 and LD90 doses, respectively (Figure 290 3B). Comparative photomicrographs (generated via Trypan blue exclusion assay) represent the 291 live and dead hemocyte cells of S. frugiperda treated with PBS and Cry1AcF, respectively 292 (Figure 3C, D). 261 Cry1AcF causes gut leakiness in S. frugiperda and S. litura 262 Presumably, orally delivered Cry1AcF catalytically disrupted the midgut architecture of test 263 insects leading to leaky gut and consequent hemolymph melanization. In order to validate this 264 hypothesis, comparative histopathology experiment of PBS- and Cry1AcF-ingested (in LD50 265 doses) S. frugiperda and S. litura midgut transverse sections was carried out at 24 h after PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 293 Cry1AcF induces immune-stimulatory activity in S. frugiperda and S. litura 293 Cry1AcF induces immune-stimulatory activity in S. frugiperda and S. litura PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 294 The disruption of hemocyte or immunocyte cell morphology due to Cry1AcF-induced toxicity is 295 indicative of Cry1AcF immune-stimulatory activity in test insects. Additionally, larval 296 melanization due to Cry1AcF catalytic activity exemplifies the elevated PO enzyme activity in 297 the hemolymph resulting from the conversion of hemocyte-bound proPO to PO. In view of this, 298 hemolymph PO activity was measured in the test insects at 12 (in order to detect the early 299 responses in host insect) and 24 h after Cry1AcF ingestion. Compared to PBS (0.05 300 OD490/min/mg protein), a significantly (F (2, 27) = 19.24, P = 0.004) higher amount of PO 301 induction (0.42-0.68 OD490/min/mg protein) was documented in S. frugiperda larvae ingested 302 with LD50 and LD90 doses of the toxin, at 12 h after inoculation (Figure 4A). At 24h, PO activity 303 further increased (F (2, 27) = 17.56, P = 0.003) to 0.62-0.92 OD490/min/mg protein in S. 304 frugiperda (Figure 4B). An identical trend was observed in S. litura larvae (Figure 4). Manuscript to be reviewed 324 proteins may play crucial role during Cry1AcF intoxication process in S. frugiperda and S. 325 litura. However, the possibility that the upregulation of these receptor genes was an indirect 326 response to the action of the Cry toxin in the midgut tissue cannot be dismissed because of our 327 inability of using a non-receptor midgut protein as a control. 324 proteins may play crucial role during Cry1AcF intoxication process in S. frugiperda and S. 325 litura. However, the possibility that the upregulation of these receptor genes was an indirect 326 response to the action of the Cry toxin in the midgut tissue cannot be dismissed because of our 327 inability of using a non-receptor midgut protein as a control. 305 Cry1AcF intoxication altered the expression of gut receptors in S. frugiperda and S. litura 306 Initially, stage-specific and tissue-specific expression profiles of different Cry receptor-encoding 307 genes was analyzed in S. frugiperda and S. litura via RT-qPCR. Transcripts of CAD, ABCC2, 308 ALP1 and APN receptor were most abundantly expressed (P < 0.01) in the fourth- and fifth- 309 instar stage compared to other life stages of S. frugiperda (Supplementary Figure 1). All the 310 receptor gene mRNAs were greatly upregulated (P < 0.01) in the midgut tissues than other body 311 parts including foregut, hindgut, Malpighian tubules, head and fat body of S. frugiperda fourth- 312 instar stage (Supplementary Figure 2). Likewise, CAD, ABCC2, ALP1 and APN transcripts 313 were abundantly expressed (P < 0.01) in both fourth- and fifth-instar stages of S. litura 314 (Supplementary Figure 3). All these receptor mRNAs were highly upregulated in the midgut 315 tissues compared to other body parts of S. litura (Supplementary Figure 4). 316 Intriguingly, CAD, ABCC2, ALP1 and APN receptor-encoding genes were significantly (P < 317 0.01) overexpressed in the midgut tissues in S. frugiperda fourth-instar larvae at 12 h after oral 318 administration of Cry1AcF toxin in LD50 dose, compared to the expression levels of identical 319 genes in PBS ingested larvae. At 24 h after toxin ingestion, expression level of these genes were 320 further upregulated by several folds (P < 0.001) compared to their expression level during 12 h 321 time period (Figure 5A). A similar trend was observed with S. litura fourth-instar larvae. Most 322 importantly, CAD transcripts were greatly expressed (P < 0.001) in S. litura at both 12 and 24 h 323 after Cry1AcF ingestion (Figure 5B). Herein, we assume that one or few of these receptor PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 354 frugiperda CAD, ABCC2, ALP1 and APN shared 98, 99, 99 and 80% identity in the dsRNA- 355 binding regions with homologous transcripts XM_035584870, OL955484, XM_035587634 and 356 XM_022979826, respectively (Figure 6A). S. litura CAD, ALP1 and APN shared 99, 100 and 357 99% identity in the dsRNA-binding regions with homologous transcripts XM_022970523, 358 XM_022967284 and XM_022969246, respectively (Figure 6B). This suggests that RNAi of 359 receptor-encoding genes might have silenced their allelic variants as well. Nevertheless, this 360 could not be proved experimentally because of the limitations associated with designing specific 361 primers, which would distinguish the two highly homologous gene transcripts. 362 RNAi-induced downregulation of CAD, ABCC2, ALP1 and APN transcripts in the midgut of S. 363 frugiperda or S. litura did not confer any off-target effect on larval development and 364 metamorphosis process as evidenced by the percent pupation and adult emergence data obtained 365 at 7-10 days after dsRNA ingestion (Supplementary Figure 7). 362 RNAi-induced downregulation of CAD, ABCC2, ALP1 and APN transcripts in the midgut of S. 363 frugiperda or S. litura did not confer any off-target effect on larval development and 364 metamorphosis process as evidenced by the percent pupation and adult emergence data obtained 365 at 7-10 days after dsRNA ingestion (Supplementary Figure 7). 366 Post 24 h of dsRNA ingestion, larvae were force-fed with Cry1AcF toxin in LD50 and LD90 367 doses followed by incubated in artificial diet. After another 24h, larval mortality data was 368 recorded. Interestingly, in CAD and ABCC2 silenced S. frugiperda, Cry1AcF (in both sub-lethal 369 and lethal doses)-induced toxicity did not differ significantly (P > 0.01) with that of PBS- or 370 GFP dsRNA-treated larvae (Figure 7A). On the contrary, compared to control, Cry1AcF (in both 371 LD50 and LD90 doses)-induced mortality in ALP1 and APN dsRNA pretreated S. frugiperda were 372 significantly (P < 0.01) reduced by 42.85-43.39% and 46.15-49.06%, respectively (Figure 7A). 373 However, in ALP1 and APN silenced S. litura, Cry1AcF (in LD50 and LD90 doses)-induced 374 toxicity did not differ significantly (P > 0.01) with that of PBS- or GFP dsRNA-treated larvae 375 (Figure 7B). Compared to control, a marginal yet significant (P < 0.01) reduction (28.57 and 376 21.05% in LD50 and LD90 doses, respectively) in S. litura mortality was documented when larvae 377 were pretreated with ABCC2 dsRNAs (Figure 7B). When CAD transcripts were knocked down 378 in S. 328 RNAi of gut receptor-encoding genes altered S. frugiperda and S. litura susceptibility to 329 Cry1AcF 330 Target dsRNA molecules (ideally 400-500 bp long sequences are readily processed by RNaseIII 331 or Dicer enzyme) from the coding sequences of S. frugiperda (CAD: 411 bp, ABCC2: 406 bp, 332 ALP1: 440 bp, APN: 410 bp) and S. litura (CAD: 427 bp, ABCC2: 447 bp, ALP1: 419 bp, APN: 333 442 bp) were designed according to the prediction of greater siRNA generation probabilities in 334 the targeted region compared to untargeted stretches in the same sequence. Next, possible RNAi 335 off-target effect was analyzed by using our target dsRNA sequences as query against Drosophila 336 melanogaster siRNA database in DsCheck server (http://dscheck.rnai.jp/). Results indicated 337 minimal homology of D. melanogaster siRNAs with that of siRNAs generated from dsRNAs of 338 CAD, ABCC2, ALP1 and APN (data not shown). DsRNAs (cloned in L4440 vector) were 339 expressed via E. coli HT115 (RNase III deficient) cells since ingestion of this recombinant 340 bacteria can produce intact dsRNA molecules in the insect gut. 341 CAD, ABCC2, ALP1 and APN expression in the midgut tissues of S. frugiperda were 342 considerably (P < 0.01) reduced by 75, 61, 79 and 65% in CAD, ABCC2, ALP1 and APN 343 dsRNA-ingested larvae, respectively, compared to their expression in PBS-ingested larvae at 24 344 h after dsRNA treatment (Supplementary Figure 5; fold change in expression of a target gene in 345 dsRNA-treated larvae was subtracted from that in control larvae and converted into percentage 346 values). Similarly, CAD, ABCC2, ALP1 and APN expression in S. litura gut were markedly (P 347 < 0.01) downregulated by 76, 58, 68 and 72% in CAD, ABCC2, ALP1 and APN dsRNA- 348 ingested larvae, respectively, compared to control (Supplementary Figure 6). Ingestion of GFP 349 dsRNA did not change (P > 0.01) the expression of receptor genes mRNAs in either of the insect 350 species. In addition, transcript abundance of CAD was unchanged (P > 0.01) in ABCC2, ALP1 351 and APN dsRNA-treated insects. Vice versa was observed with other receptor mRNAs as well in 352 either of the insect species (Supplementary Figures 5, 6). These results has indicated the target- 353 specific induced silencing of receptor-encoding genes in the midgut tissues. However, S. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 384 Discussion 385 Chimeric toxins provide an important armory towards the diversification of Cry protein 386 repertoire especially when insect pests are constantly developing resistance against narrow- 387 spectrum Cry bio-pesticides or Bt crops globally (Zafar et al. 2020; Chen et al. 2021). A 388 chimeric toxin Cry1AcF (generated by swapping domains of Cry1Ac and Cry1F; Rathinam et al. 389 2017, 2019) was transgenically expressed in different crop plants and exhibited toxicity against 390 lepidopteran pests (Keshamma et al. 2012; Ramu et al. 2012; Muralimohan et al. 2020; 391 Ramkumar et al. 2020). The synergistic effect of Cry1Ac and Cry1F in 1:1 ratio augmented the 392 insecticidal efficacy of chimeric toxin by 26 times (Ramu et al. 2012). In the current study, a 393 detailed in vivo pathogenesis assay of Cry1AcF was carried out in two notorious noctuid pests S. 394 frugiperda (an invasive pest showing increased incidence under global climate change) and S. 395 litura. Oral ingestion of Cry1AcF caused greater toxicity in the fourth-instar larvae of S. 396 frugiperda (LD50: 30 ng/larva; LD90: 100 ng/larva) than S. litura (LD50: 40 ng/larva; LD90: 120 397 ng/larva) along with phenotypic alterations such as dead-like posture with extended prolegs and 398 hemolymph melanization at 24 h post inoculation. 399 Histopathology of the midgut transverse sections showed Cry1AcF-induced extensive gut 400 damage in both S. frugiperda and S. litura fourth-instar larvae. Compared to control larvae, 401 toxin-ingested larvae exhibited disintegrated and sloughed midgut epithelial tissue into the 402 midgut lumen followed by rupture of basement membrane and visceral muscle layer (at midgut- 403 homecoel interface) putatively leading to leaky gut. An analogous histopathology data was 404 previously documented in our laboratory when Cry1AcF toxin was orally delivered into the 405 greater wax moth G. mellonella larvae (Dutta et al. 2022a). Similarly, an orally active bacterial 406 toxin TcaB (characterized from the entomopathogenic bacterium Photorhabdus akhurstii) caused 407 lysis of the midgut epithelial cells followed by shedding of epithelium into the lumen leading to 408 compromised gut homeostasis in S. frugiperda, S. litura and H. armigera fourth-instar larvae 409 (Dutta et al. 2021). Because δ-endotoxin and vegetative insecticidal protein (Vip3A) of B. 410 thuringiensis exhibit similar gut-active nature (Adang et al. 2014; Chakrabarty et al. 2022), the 411 pathogenesis process of Cry1AcF can be correlated with those toxins. Manuscript to be reviewed litura, a greatest (P < 0.01) reduction (55.36 and 52.63% in LD50 and LD90 doses, 379 respectively) in larval mortality was documented in comparison to control larvae (Figure 7B). 380 Taken together, our data putatively indicates ALP1/APN and CAD as potential receptors for 381 Cry1AcF intoxication in S. frugiperda and S. litura, respectively, because induced suppression of 382 these receptor-encoding genes disrupted the larval susceptibility to Cry1AcF at both LD50 and 383 LD90 doses. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 414 hemocyte cells and trigger apoptosis and cytolysis in order to modulate innate immune responses 415 of host insects (Whitten and Coates 2017). In the current study, at 24 h after oral ingestion, 416 Cry1AcF elicited a sharp decline in total circulatory hemocyte numbers as well as hemocyte 417 viability in S. frugiperda and S. litura fourth-instar larvae, compared to control larvae. Likewise, 418 a substantial reduction in hemocyte counts because of Cry toxin-induced gut permeability was 419 demonstrated in different insects (Ericsson et al. 2009; Abd El-Aziz and Awad 2010; Broderick 420 et al. 2010; Manachini et al. 2011; Grizanova et al. 2014). 414 hemocyte cells and trigger apoptosis and cytolysis in order to modulate innate immune responses 415 of host insects (Whitten and Coates 2017). In the current study, at 24 h after oral ingestion, 416 Cry1AcF elicited a sharp decline in total circulatory hemocyte numbers as well as hemocyte 417 viability in S. frugiperda and S. litura fourth-instar larvae, compared to control larvae. Likewise, 418 a substantial reduction in hemocyte counts because of Cry toxin-induced gut permeability was 419 demonstrated in different insects (Ericsson et al. 2009; Abd El-Aziz and Awad 2010; Broderick 420 et al. 2010; Manachini et al. 2011; Grizanova et al. 2014). 414 hemocyte cells and trigger apoptosis and cytolysis in order to modulate innate immune responses 415 of host insects (Whitten and Coates 2017). In the current study, at 24 h after oral ingestion, 416 Cry1AcF elicited a sharp decline in total circulatory hemocyte numbers as well as hemocyte 417 viability in S. frugiperda and S. litura fourth-instar larvae, compared to control larvae. Likewise, 418 a substantial reduction in hemocyte counts because of Cry toxin-induced gut permeability was 419 demonstrated in different insects (Ericsson et al. 2009; Abd El-Aziz and Awad 2010; Broderick 420 et al. 2010; Manachini et al. 2011; Grizanova et al. 2014). 421 When challenge inoculated with toxin-like biomolecules, insects exhibit humoral immune 422 response such as hemolymph melanization. This is resulted due to increased phenoloxidase 423 enzyme activity in the hemolymph because hemocyte-bound pro-phenoloxidases are converted 424 into phenoloxidases (Whitten and Coates 2017). In the present study, Cry1AcF ingestion 425 markedly elevated the hemolymph phenoloxidase activity in S. frugiperda and S. litura fourth- 426 instar larvae, compared to control larvae. 384 Discussion 412 Upon reaching the body cavity via leaky gut, Cry1AcF might have targeted the hemocyte or 413 immunocyte cells in order to cause toxicity. Toxin-like biomolecules frequently target the 412 Upon reaching the body cavity via leaky gut, Cry1AcF might have targeted the hemocyte or 413 immunocyte cells in order to cause toxicity. Toxin-like biomolecules frequently target the PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed 444 S. litura is dependent on binding to these receptors or not by deploying RNAi-mediated 445 knockdown of individual receptor-encoding genes. Initially, developmental stage-and tissue- 446 specific transcription patterns of CAD, ABCC2, ALP1 and APN in test insects was analyzed by 447 RT-qPCR. All the receptor gene transcripts were greatly expressed in both fourth- and fifth- 448 instar stages of S. frugiperda and S. litura. Additionally, all the receptor mRNAs were most 449 abundantly expressed in the midgut tissues of S. frugiperda and S. litura fourth-instar larvae. In 450 our earlier study, S. frugiperda and S. litura fourth-instar larval stages were found to be highly 451 vulnerable to a bacterial toxin TcaB (Dutta et al. 2021). 452 Oral ingestion of bacterially-expressed dsRNAs of CAD, ABCC2, ALP1 and APN 453 downregulated (compared to control) the transcription of corresponding receptor mRNAs by 75- 454 76, 58-61, 68-79 and 65-72%, respectively, in the midgut tissue of S. frugiperda and S. litura 455 fourth-instar larvae. Probability of any cross knockdown effect among the receptor mRNAs was 456 non-existent as the sequence similarly between targeted dsRNA sequences of CAD, ABCC2, 457 ALP1 and APN within the same test insect species was negligible (data not shown). In addition, 458 silencing of receptor genes did not cause any negative effect on the growth and metamorphosis 459 of either of the test insects. 452 Oral ingestion of bacterially-expressed dsRNAs of CAD, ABCC2, ALP1 and APN 460 Target-specific induced downregulation of receptor mRNAs variably altered the insect 461 susceptibility to Cry1AcF toxin in our study. For example, the susceptibility of ALP1 and APN 462 dsRNA-treated S. frugiperda larvae was decreased (compared to control) by 42.85-43.39 and 463 46.15-49.06%, respectively, when ingested with Cry1AcF toxin in LD50 and LD90 doses. On the 464 contrary, susceptibility of CAD and ABCC2 dsRNA-treated S. frugiperda was alike of control 465 larvae when ingested with Cry1AcF in lethal and sub-lethal doses. In ALP1 and APN dsRNA- 466 treated S. litura, larval susceptibility was alike of control when ingested with Cry1AcF in LD50 467 and LD90 doses. Conversely, susceptibility of CAD dsRNA-treated S. litura was decreased 468 (compared to control) by 55.36 and 52.63%, upon ingestion with Cry1AcF in LD50 and LD90 469 doses, respectively. Additionally, S. litura larvae pre-treated with ABCC2 dsRNA showed a 470 meagre yet significant reduction (21.05-28.57%) in insect vulnerability to Cry1AcF (in LD50 and 471 LD90 doses), compared to control. Manuscript to be reviewed Intriguingly, phenoloxidase activity was initiated 427 during early pathogenesis (12 h after Cry1AcF inoculation) and was further increased to 428 manifold during late pathogenesis (24 h). Our results suggest that, Cry1AcF can be used as an 429 immunomodulatory or immune-stimulatory agent in insect defense response studies. Notably, a 430 three-day sustaining Cry-induced increased hemolymph phenoloxidase activity was documented 431 in G. mellonella (Grizanova et al. 2014). Similar data was documented in H. armigera and 432 Trichoplusia ni upon challenge inoculation with different Cry toxins (Rahman et al., 2004; Ma et 433 al., 2005; Ayres and Schneider, 2008). Cerenius et al. (2008) speculated that elevated 434 phenoloxidase activity is resulted due to induced gut repair process, which limits the toxin 435 escape into body cavity through Cry-elicited perforated gut. Nevertheless, other studies argued 436 that Cry-elicited immunomodulation prevents host insect resistance to further Cry intoxication in 437 the intestine (Kwon and Kim, 2007; Broderick et al., 2010; Richards and Dani, 2010). The 438 initiation of gut repair process was not observed in either of the test insects ingested with 439 Cry1AcF in our histopathology experiments. 440 CAD, ABCC (transmembrane proteins), APN and ALP (tether to the epithelial cell membrane 441 via GPI-anchors) are known receptor proteins in the brush border cells that are crucial for Cry 442 binding and intoxication mechanism in the intestine (Bravo et al. 2011; Fabrick and Wu 2015; 443 Liu et al. 2021). This prompted us to analyze whether Cry1AcF intoxication in S. frugiperda and PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 482 Conclusions 483 In our study, the chimeric toxin Cry1AcF conferred potent insecticidal activity in S. frugiperda 484 and S. litura. Hemolymph melanization in the test insects was the resultant effect of Cry1AcF- 485 induced gut damage followed by cytotoxicity in the hemocyte cells. Further, RNAi-mediated 486 silencing of gut receptor encoding genes differentially altered the susceptibility of test insects to 487 Cry1AcF toxin, indicating the potential role of these receptor proteins in Cry1AcF intoxication 488 process. More specifically, our results indicated ALP1/APN and CAD/ABCC2 as the functional 489 receptor for Cry1AcF toxicity in S. frugiperda and S. litura, respectively. 483 In our study, the chimeric toxin Cry1AcF conferred potent insecticidal activity in S. frugiperda 484 and S. litura. Hemolymph melanization in the test insects was the resultant effect of Cry1AcF- 485 induced gut damage followed by cytotoxicity in the hemocyte cells. Further, RNAi-mediated 486 silencing of gut receptor encoding genes differentially altered the susceptibility of test insects to 487 Cry1AcF toxin, indicating the potential role of these receptor proteins in Cry1AcF intoxication 488 process. More specifically, our results indicated ALP1/APN and CAD/ABCC2 as the functional 489 receptor for Cry1AcF toxicity in S. frugiperda and S. litura, respectively. Manuscript to be reviewed 474 formation model of Cry mechanism of action. In this model, Domain I of Cry protein attaches 475 into the brush border cell membrane via its hydrophobic helical hairpin, Domain II identifies 476 receptor molecules and Domain III aids in receptor binding and pore formation (Pacheco et al. 477 2009; Pardo-López et al. 2013; Adang et al. 2014; Jin et al. 2021; Jurat-Fuentas et al. 2021). In 478 signaling pathway model, Cry binding to CAD receptor triggers adenylate cyclase activation that 479 stimulates increased cAMP and protein kinase synthesis. This leads to an ion channel (formed in 480 the brush border cell membrane)-dependent cellular signaling cascading effect, which ultimately 481 causes cell death (Zhang et al. 2006; Castella et al. 2019). 490 Acknowledgements 491 We acknowledge Dr. S. Subramanian and Dr. D. Sagar, Division of Entomology, ICAR-Indian 492 Agricultural Research Institute for providing us the laboratory space for rearing insects and 493 conducting RNAi experiments. 491 We acknowledge Dr. S. Subramanian and Dr. D. Sagar, Division of Entomology, ICAR-Indian 492 Agricultural Research Institute for providing us the laboratory space for rearing insects and 491 We acknowledge Dr. S. Subramanian and Dr. D. Sagar, Division of Entomology, ICAR-Indian 491 We acknowledge Dr. S. Subramanian and Dr. D. Sagar, Division of Entomology, ICAR-Indian 492 Agricultural Research Institute for providing us the laboratory space for rearing insects and 493 conducting RNAi experiments. 492 Agricultural Research Institute for providing us the laboratory space for rearing insects and 493 d ti RNAi i t 492 Agricultural Research Institute for providing us the laboratory space for rea Manuscript to be reviewed Taken together, our results indicate ALP1/APN and 472 CAD/ABCC2 as the functional receptor for Cry1AcF toxicity in S. frugiperda and S. litura, 473 respectively. Notably, either of the ALP, APN and ABCC receptors are involved in pore PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) 494 References: 495 Abd El-Aziz, N.M., Awad, H.H. (2010). Changes in the haemocytes of Agrotis ipsilon larvae 496 (Lepidoptera: Noctuidae) in relation to dimilin and Bacillus thuringiensis infections. Micron. 41, 497 203–209. 95 Abd El-Aziz, N.M., Awad, H.H. (2010). 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Pest Management Science 76(5), 1902-1911. 608 Assessment of Pigeonpea (Cajanus cajan L.) transgenics expressing Bt ICP 609 Cry1AcF under nethouse containment implicated an effective control against herbivory by l ( b ) i ( ) 609 Cry1AcF under nethouse containment implicated an effective control again 611 Ramu, S.V., Rohini, S., Keshavareddy, G., Gowri Neelima, M., Shanmugam 611 Ramu, S.V., Rohini, S., Keshavareddy, G., Gowri Neelima, M., Shanmugam, N.B., Kumar, 612 A.R.V., et al. (2012). Expression of a synthetic cry1AcF gene in transgenic Pigeon pea confers 613 resistance to Helicoverpa armigera. Journal of Applied Entomology 136(9), 675-687. 612 A.R.V., et al. (2012). Expression of a synthetic cry1AcF gene in transgenic Pigeon pea confers 613 resistance to Helicoverpa armigera. Journal of Applied Entomology 136(9), 675-687. 612 A.R.V., et al. (2012). Expression of a synthetic cry1AcF gene in transgenic Pigeon pea confers i l l f li d l ( ) 614 Rathinam M, Kesiraju K, Singh S, Thimmegowda V, Rai V, Pattanayak D, Sreevathsa R (2019). 615 Molecular interaction-based exploration of the broad spectrum efficacy of a Bacillus 616 thuringiensis insecticidal chimeric protein, Cry1AcF. Toxins 11:143. 617 Rathinam, M., Singh, S., Pattanayak, D., Sreevathsa, R. (2017). Comprehensive in silico 618 allergenicity assessment of novel protein engineered chimeric cry proteins for safe deployment in 619 crops. BMC Biotechnology, 17, 64. 620 Richards, E.H., Dani, M.P. (2010). A recombinant immunosuppressive protein from Pimpla 621 hypochondriaca (rVPr1) increases the susceptibility of Lacanobia oleracea and Mamestra 622 brassicae larvae to Bacillus thuringiensis. J. Invertebr. Pathol. 104, 51–57. 620 Richards, E.H., Dani, M.P. (2010). Manuscript to be reviewed P., Kesiraju, K., Pattanayak, D., Ananda Kumar, P., Kasturi, K., & 592 Sreevathsa, R. (2020). Molecular stacking of two codon-modified genes encoding Bt insecticidal 593 proteins, Cry1AcF and Cry2Aa for management of resistance development in Helicoverpa 594 armigera. Journal of Plant Biochemistry and Biotechnology 29(3), 518-527. 595 Pacheco S, Gómez I, Gill SS, Bravo A, Soberón M. (2009). Enhancement of insecticidal activity 596 of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with 597 oligomer formation. Peptides 30:583–588. 598 Pardo-López, L., Sóberon, M., Bravo, A. (2013). Bacillus thuringiensis insecticidal three-domain 599 Cry toxins: mode of action, insect resistance and consequences for crop protection. FEMS 600 Microbiol. Rev. 37:3-22. 601 Phani, V., Khan, M.R., Dutta, T.K. (2021). Plant-parasitic nematodes as a potential threat to 602 protected agriculture: current status and management options. Crop Prot 144:105573. doi: 603 10.1016/j.cropro.2021.105573. 601 Phani, V., Khan, M.R., Dutta, T.K. (2021). Plant-parasitic nematodes as a potential threat to 602 protected agriculture: current status and management options. Crop Prot 144:105573. doi: 603 10.1016/j.cropro.2021.105573. p g g p p 603 10.1016/j.cropro.2021.105573. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed A recombinant immunosuppressive pro 620 Richards, E.H., Dani, M.P. (2010). A recombinant immunosuppressive protein from Pimpla 621 hypochondriaca (rVPr1) increases the susceptibility of Lacanobia oleracea and Mamestra 621 hypochondriaca (rVPr1) increases the susceptibility of Lacanobia oleracea 621 hypochondriaca (rVPr1) increases the susceptibility of Lacanobia oleracea and Mamestra 622 brassicae larvae to Bacillus thuringiensis. J. Invertebr. Pathol. 104, 51–57. 623 Santhoshkumar K, Mathur C, Mandal A, Dutta TK (2021). A toxin complex protein from 624 Photorhabdus akhurstii conferred oral insecticidal activity against Galleria mellonella by 625 targeting the midgut epithelium. Microbiol Res. 242:126642. 626 Shankhu PY, Mathur C, Mandal A, Sagar D, Somvanshi VS, Dutta TK (2020). Txp40, a protein 627 from Photorhabdus akhurstii conferred potent insecticidal activity against the larvae of 628 Helicoverpa armigera, Spodoptera litura and S. exigua. Pest Manag Sci. 76:2004–2014. 626 Shankhu PY, Mathur C, Mandal A, Sagar D, Somvanshi VS, Dutta TK (2020). Txp40, a protein 627 from Photorhabdus akhurstii conferred potent insecticidal activity against the larvae of 628 Helicoverpa armigera Spodoptera litura and S exigua Pest Manag Sci 76:2004 2014 , , , g , , ( ) p , p 627 from Photorhabdus akhurstii conferred potent insecticidal activity against the larvae of 28 Helicoverpa armigera, Spodoptera litura and S. exigua. Pest Manag Sci. 76:2 629 Suby, S.B., Soujanya, P.L., Yadava, P., Patil, J., Subaharan, K., Prasad, G.S., et al. (2020). 630 Invasion of fall armyworm (Spodoptera frugiperda) in India: nature, distribution, management 631 and potential impact. Curr. Sci. 119, 44–51. 630 Invasion of fall armyworm (Spodoptera frugiperda) in India: nature, distribution, management 631 and potential impact. Curr. Sci. 119, 44–51. 632 Tabashnik BE, Brévault T, Carrière Y (2013). Insect resistance to Bt crops: lessons from the first 633 billion acres. Nat Biotechnol 31:510–521. 634 Tabashnik, B.E., Carrière, Y. (2017). Surge in insect resistance to transgenic crops and prospects 635 for sustainability. Nat Biotechnol. 35, 926–935. 634 Tabashnik, B.E., Carrière, Y. (2017). Surge in insect resistance to transgeni PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Manuscript to be reviewed Table 1(on next page) Manuscript to be reviewed 636 Tajne, S., Boddupally, D., Sadumpati, V., Vudem, D.R., Khareedu, V.R. (2014). Synthetic 637 fusion-protein containing domains of Bt Cry1Ac and Allium sativum lectin (ASAL) conferred 638 enhanced insecticidal activity against major lepidopteran pests. J. Biotech. 171, 71–75. 637 fusion-protein containing domains of Bt Cry1Ac and Allium sativum lectin (ASAL) conferred 638 enhanced insecticidal activity against major lepidopteran pests. J. Biotech. 171, 71–75. 639 Whitten, M.M., Coates, C.J. (2017). Re‐evaluation of insect melanogenesis research: views from 640 the dark side. Pigment Cell Melanoma Res. 30, 386-401. 641 Zafar, M.M., Razzaq, A., Farooq, M.A., Rehman, A., Firdous, H., Shakeel, A., et al. (2020). 642 Insect resistance management in Bacillus thuringiensis cotton by MGPS (multiple genes 643 pyramiding and silencing). Journal of Cotton Research 3, 33. 644 Zghal, R.Z., Elleuch, J., Ali, M.B., Darriet, F., Rebaï, A., Chandre, F., Tounsi, S. (2017). 645 Towards novel Cry toxins with enhanced toxicity/broader: A new chimeric Cry4Ba/Cry1Ac 646 toxin. Appl. Microbiol. Biotech. 101, 113–122. 647 Zhang, X., Candas, M., Griko, N.B., Taussig, R., Bulla, LA Jr. (2006). A mechanism of cell 648 death involving an adenylyl cyclase/PKA signaling pathway is induced by the Cry1Ab toxin of 649 Bacillus thuringiensis. Proc. Natl. Acad. Sci. U.S.A. 103, 9897–9902. 649 Bacillus thuringiensis. Proc. Natl. Acad. Sci. U.S.A. 103, 9897–9902. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Table 1 The LD50 and LD90 values of Cry1AcF toxin in S. frugiperda and S. litura fourth-instar larvae at 24 h after oral ingestion. A known bacterial toxin TcaB was used as the reference. Values were determined by Probit analysis. Numbers in parentheses represent 95% conﬁdence interval. R 2 value indicates the closeness of data to the ﬁtted regression line. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed nd LD90 values of Cry1AcF toxin in S. frugiperda and S. litura fourth-instar larvae at 24 h after oral ingestion. A n TcaB was used as the reference. Values were determined by Probit analysis. Numbers in parentheses represent val. R2 value indicates the closeness of data to the fitted regression line. LD50 LD90 R2 value Cry1AcF (ng/larva) 29.78 (15.55 – 39.54) 99.77 (78.89 – 121.78) 0.922 TcaB (ng/larva) 53.89 (41.33 – 69.72) 148.49 (127.85-170.43) 0.918 Cry1AcF (ng/g)* 53.60 179.59 - TcaB (ng/g)* 97.00 267.28 - Cry1AcF (ng/larva) 38.76 (22.88 – 51.67) 117.32 (93.66 – 133.65) 0.903 TcaB (ng/larva) 69.86 (55.98 – 88.39) 172.19 (146.75 – 191.48) 0.925 Cry1AcF (ng/g)* 69.77 211.18 - TcaB (ng/g)* 125.75 309.94 - mass during inoculation was ~ 0.45 g. 1 Table 1. The LD50 and LD90 values of Cry1AcF toxin in S. frugiperda and S. litura fourth-instar larvae at 24 h after oral ingestion. A 2 known bacterial toxin TcaB was used as the reference. Values were determined by Probit analysis. Numbers in parentheses represent 3 95% confidence interval. R2 value indicates the closeness of data to the fitted regression line. 1 Table 1. The LD50 and LD90 values of Cry1AcF toxin in S. frugiperda and S. litura fourth-instar larvae at 24 h after oral ingestion. A 2 known bacterial toxin TcaB was used as the reference. Values were determined by Probit analysis. Numbers in parentheses represent 3 95% confidence interval. R2 value indicates the closeness of data to the fitted regression line. LD50 LD90 R2 value S. frugiperda Cry1AcF (ng/larva) 29.78 (15.55 – 39.54) 99.77 (78.89 – 121.78) 0.922 TcaB (ng/larva) 53.89 (41.33 – 69.72) 148.49 (127.85-170.43) 0.918 Cry1AcF (ng/g)* 53.60 179.59 - TcaB (ng/g)* 97.00 267.28 - S. litura Cry1AcF (ng/larva) 38.76 (22.88 – 51.67) 117.32 (93.66 – 133.65) 0.903 TcaB (ng/larva) 69.86 (55.98 – 88.39) 172.19 (146.75 – 191.48) 0.925 Cry1AcF (ng/g)* 69.77 211.18 - TcaB (ng/g)* 125.75 309.94 - 4 * The average larval mass during inoculation was ~ 0.45 g. Manuscript to be reviewed Table 1 4 * The average larval mass during inoculation was ~ 0.45 g. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Figure 1 Insecticidal activity of Cry1AcF toxin in S. frugiperda and S. litura. (A) Oral ingestion of the toxin using hypodermic needle in starved S. frugiperda fourth-instar larvae. (B) At 24 h after inoculation, toxin (150 ng)-treated moribund (M) larvae attained dead-like posture with extended prolegs and hemolymph melanization compared to normal phenotypes of PBS- ingested control (L) larvae (scale bar – 0.5 cm). Dose-response curves depict the percent survival of S. frugiperda (C) and S. litura (D) at 24 h after toxin ingestion. TcaB and PBS were used as the positive and negative control, respectively. X- and Y-axis represent toxin dose and percent larval survival, respectively. Treatments (mean ± SE, n = 50) with diﬀerent letters are signiﬁcantly diﬀerent at P < 0.01, Tukey’s HSD test. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Figure 2 Cry1AcF negatively altered the gut homeostasis of S. frugiperda and S. litura fourth-instar larvae. Histopathology of the midgut transverse sections of S. frugiperda treated with PBS (A), Cry1AcF (B) and S. litura treated with PBS (C), Cry1AcF (D) at 24 h after inoculation. Cry1AcF was administered with LD50 doses. Asterisks indicate disintegration of epithelial cells (EC) followed by sloughing oﬀ into the gut lumen (GL). VM – visceral muscle, Nu – Nucleus. Scale bar = 50 μm. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Figure 3 Eﬀect of Cry1AcF toxin on the total circulatory hemocytes of S. frugiperda and S. litura fourth-instar larvae at LD50 and LD90 doses. (A) Y-axis indicates hemocyte counts (×10 5) per ml of hemocoel extracted at 24 h after toxin ingestion. (B) Y-axis indicates percent viable cells (determined by Trypan blue staining assay) at 24 h after toxin ingestion. PBS was used as the control. Treatments (mean ± SE, n = 10) with diﬀerent letters are signiﬁcantly diﬀerent at P < 0.01, Tukey’s HSD test. Comparative photomicrographs show live cells (prevent dye invasion) in PBS ingested S. frugiperda larvae and dead cells (stain dark blue because of disintegrated membrane) in Cry1AcF ingested S. frugiperda larvae. Scale bar = 10 μm. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Figure 4 Hemolymph phenoloxidase (PO) enzyme activity in S. frugiperda and S. litura fourth-instar larvae upon ingestion of Cry1AcF in LD50 and LD90 doses. PO activity was measured at 12 (A) and 24 (B) h after toxin administration. PBS was used as the control. PO activity was measured using L-DOPA as substrate and change in absorbance (due to melanin synthesis) after 30 min was recorded at 490 nm. PO activity was expressed as OD490 per minute per mg protein. PBS + L-DOPA was used as blank and its absorbance reading (at 490 nm) was subtracted from each sample. Treatments (mean ± SE, n = 10) with diﬀerent letters are signiﬁcantly diﬀerent at P < 0.01, Tukey’s HSD test. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Figure 5 Relative expression of receptor protein encoding genes in the midgut of S. frugiperda (A) and S. litura (B) fourth-instar larvae at 12 and 24 h after Cry1AcF ingestion (in LD50 doses). Asterisk (P < 0.01, *P < 0.001; Tukey’s HSD test) is indicative of signiﬁcant diﬀerential expression of target gene mRNAs compared to their baseline expression (fold change values set at 1) in insects ingested with PBS. Gene expression was normalized using endogenous reference genes of S. frugiperda (β-actin and GAPDH) and S. litura (rps3). Each bar represents the mean fold change value ± SE of RT-qPCR runs in ﬁve biological and three technical replicates. technical replicates. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Manuscript to be reviewed Manuscript to be reviewed Manuscript to be reviewed Figure 6 The dsRNA binding sites (indicated by arrows and perpendicular lines) in the coding sequences of S. frugiperda (A) and S. litura (B) CAD, ABCC2, ALP1 and APN are shown. Numbers indicate the sequence coordinates. The homologous transcripts (indicated by grey boxes) that were aligned with the target receptors are shown. Perpendicular lines indicate percent identity in the dsRNA target region. NCBI Genebank identiﬁers for diﬀerent transcripts are listed at the end. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) Figure 7 Figure 7 RNAi-mediated silencing (si) of Cry receptor encoding genes diﬀerentially altered the susceptibility of S. frugiperda (A) and S. litura (B) to Cry1AcF toxin. Initially, fourth-instar larvae were orally ingested with target receptor dsRNA-expressing E. coli HT115 cells. At 24 h of dsRNA ingestion, LD50 (30 and 40 ng per larva for S. frugiperda and S. litura, respectively) and LD90 (100 and 120 ng per larva for S. frugiperda and S. litura, respectively) doses of Cry1AcF were orally administered. After another 24 h percent mortality data was recorded. Diﬀerent letters indicate treatments (mean ± SE) are signiﬁcantly diﬀerent at P < 0.01, Tukey’s HSD test, n = 50. GFP dsRNA and PBS were used as the non-native and negative control, respectively. PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022) PeerJ reviewing PDF \| (2022:09:77909:2:0:NEW 15 Dec 2022)
https://openalex.org/W2591899297	https://www.mdpi.com/2073-4441/9/3/197/pdf?version=1489052871	English	null	Establishment and Application of Wetlands Ecosystem Services and Sustainable Ecological Evaluation Indicators	Water	2,017	cc-by	7,110	Han-Shen Chen 1,2 Han Shen Chen 1 School of Health Diet and Industry Management, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan; allen9750@yahoo.com.tw; Tel.: +886-4-2473-0022 2 Department of Medical Management, Chung Shan Medical University Hospital, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan 1 School of Health Diet and Industry Management, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan; allen9750@yahoo.com.tw; Tel.: +886-4-2473-0022 2 Department of Medical Management, Chung Shan Medical University Hospital, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan 1 School of Health Diet and Industry Management, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan; allen9750@yahoo.com.tw; Tel.: +886-4-2473-0022 2 Department of Medical Management, Chung Shan Medical University Hospital, No. 110, Sec. 1, Jianguo N. Rd., Taichung City 40201, Taiwan Academic Editor: Peter L. M. Goethals Received: 8 November 2016; Accepted: 7 March 2017; Published: 8 March 2017 Academic Editor: Peter L. M. Goethals Received: 8 November 2016; Accepted: 7 March 2017; Published: 8 March 2017 Received: 8 November 2016; Accepted: 7 March 2017; Published: 8 March 2017 Abstract: Gaomei wetlands are national Taiwanese coastal wetlands. Over the past few years, they have grown into an important water bird habitat and popular bird-watching location. However, the rapid growth in tourism has begun to affect the environmental quality in the Gaomei wetlands. This study combined ecosystem services (ES) and ecological footprint (EF) assessments to evaluate the sustainability status according to the features of each ecosystem service for the different Gaomei wetlands land uses. The results found that (a) the total Gaomei wetlands ecosystem service value increased from 59.24 million TWD in 2008 to 98.10 million TWD in 2015, and the ecosystem service function was continuously improving; (b) the EF increased by 56.12% over 8 years; and (c) there was a negative growth rate of 106.54% in the ecological deﬁcit (ED) in the sustainable ecological evaluation indicators (SEEI). The ecological footprint index (EFI) in 2015 was at Level 4 at 1.02, and the environmental sustainability index (ESI) was at Level 3 at 0.49. Results show that Gaomei wetlands have a low sustainability; therefore, the local, regional, and national governments need to implement regulations to strictly control the Gaomei wetlands land use. Han-Shen Chen 1,2 This study demonstrated that ES and EF theory application can give an objective guidance to decision-makers to ensure that wetlands eco-security can be maintained at safe levels. Keywords: wetlands; ecological footprint; ecosystem services; ecological security; sustainable ecological evaluation indicators (SEEI) Article Establishment and Application of Wetlands Ecosystem Services and Sustainable Ecological Evaluation Indicators Han-Shen Chen 1,2 Water 2017, 9, 197; doi:10.3390/w9030197 water water water water 1. Introduction (2011) evaluated the ecosystem service values in wetlands, mangroves, coral reefs, seagrass beds, and sandy beaches [20]. Bateman et al. (2011) explored the contribution of land use changes on ecosystem services and ecosystems [21]. Su et al. (2012) focused on four ecological zones in Hangzhou, China, to investigate the effect of landscape patterns and ecosystem service changes on urbanization [7]. To examine ecosystem services and biodiversity in Europe, Maes et al. (2012) used four supply function indicators, ﬁve adjustment function indicators, and a cultural function indicator to calculate ecosystem service values, and used average species richness and species diversity to measure biodiversity [22]. As previous research on land use and its impact on the environment has tended to focus more on exploring the single highest impact level, there has been less focus on the analysis and evaluation of the impact land-use development and the changes it has had on the natural environment. An econometric model [23,24], a statistical model [25–30] as well as a cellular automata model [31] have to date been the most commonly used evaluation models. Burkhard et al. (2013) believed that, for a more realistic ecosystem service status assessment, ecosystem services at different ecosystems, and the beneﬁts that different ecosystems and land cover types provide, should also be considered [32]. Therefore, some studies have integrated the Millennium Ecosystem Assessment (MEA) and other assessment indicators into comprehensive evaluation indices [33–35]. Nelson et al. (2009) combined land use change ecosystem services with the integrated valuation of ecosystem services and tradeoffs (InVEST) mode to explore the relationship between biodiversity competition with other ecosystem services [36]. Polasky et al. (2011) also used the InVEST mode to quantify changes in ecosystem services, biodiversity, and land use in Minnesota from 1992 to 2001, and to assess the impact of different land use change scenarios on ecosystem services and biodiversity. Using the historical development (1964–2004) in Leipzig, Germany [37], Lautenbach et al. (2011) developed regional scale indicators for different land use structure ecosystem services, such as water puriﬁcation, pollination, food production, and outdoor recreation, and calculated the systemic functions and analyzed sensitivity tests under different land use types [34]. Geneletti (2012) simulated the impact generated by different land management policies on ecosystem services in the future based on historical land use [38]. 1. Introduction As outlined in the Millennium Ecosystem Assessment of the United Nations, wetlands are one of the most threatened ecosystem in the world, with biodiversity loss being the major concern. With the continued loss and degradation of wetlands, ecological services are declining, negatively impacting human life. Factors such as climate change, rural poverty, and increased human population size have resulted in a wetlands loss of ~30%–50% in the last decade [1–3]. As well as providing ecosystem services such as ﬂood control, coastline protection, nutrient recycling, carbon sequestration, and ecotourism, wetlands support many specialized plants and animal species [4–8]. This study was conducted in the Gaomei wetlands, located in Shimizu, Taichung, Taiwan, and designated national Taiwanese wetlands in 2007 (Construction and Planning Agency Ministry of the Interior, 2007). Recently, the Gaomei wetlands have become a major bird destination as a critical winter habitat. However, human activities such as highway construction have led to signiﬁcant reductions in the sandbars and mangroves with a concurrent loss of biodiversity. Sustainable economic and societal development and reductions in the impact of tourism must be addressed when developing Water 2017, 9, 197; doi:10.3390/w9030197 www.mdpi.com/journal/water www.mdpi.com/journal/water Water 2017, 9, 197 2 of 11 natural resources. This study sought to balance resource and biodiversity conservation with sustainable management and resource development. Because the Gaomei wetlands have similar topography and ecosystems to other Taiwanese wetlands systems, the evaluation model developed in this paper could be used for similar wetlands systems. Chapin et al. (2000) suggested that ecosystem processes and biological diversity are crucial intermediaries in the overall economic and human systems’ global environment [9]. Costanza et al. (1997) deﬁned ecosystem services (ES) as ecosystems that “provide, directly or indirectly, the material and services to promote human welfare” [10]. The UN published the Millennium Ecosystem Assessment (MEA) in 2005, in which ecosystem services are divided into supply services, regulating services, cultural services, and support services [11]. Ecosystem service (ES) assessments have traditionally focused on identifying the individual monetary values for each ecosystem service [12,13]. However, the lack of theoretical frameworks has led to subjective judgments and criticisms [14–16]. The integration of deliberate and non-monetary valuation approaches to ES valuations has increasingly been advocated as a way of revealing the wider value concepts. Such methods, however, have had limited application in practice and have been mostly focused on localized case studies [17–19]. Barbier et al. 1. Introduction In summary, using analysis and prediction modes for land use change along with mode simulations, decomposition, the analysis and synthesis of the complex socio-economic factors, and the interaction processes in natural ecosystems for given land uses to determine land-use change and spatial pattern trends [39–42] have become the focus of current research trends. Sharp variabilities in the global climate have resulted in desertiﬁcation, reduced ecosystem resilience, and loss of biodiversity. The 1972 United Nations Declaration on the Human Environment and Eco-Security raised concerns related to the preservation of food and ecosystems and outlined principles for sustainable human development projects, providing a new perspective on environmental Water 2017, 9, 197 3 of 11 resources, human survival and sustainable development reviews. With the development of ecological security theory and as ecological problems became increasingly prominent, researchers began using different indicators and measurement system models or methods to evaluate the ecological security of different regional scales, thus providing early warning models that could serve as vital references [43]. As using quantitative indicators to analyze complex information increases objectivity [44,45], a three-dimensional (economy, ecology, and society) indicator system was developed to study ecological security in Western Nepal [46]. Ecological security has also been measured using the Ecological Footprint Index (EFI) and environmental carrying capacity (ECC) [47]. The Ecological Footprint (EF) Model was proposed by Rees (1992) [48], with the primary feature being its ability to compare human demands on the environment with the biosphere’s ability to regenerate resources and provide services. Wackernagel and Rees (2000) proposed that the EF magnitude was directly proportional to the environmental impact (the greater the EF, the greater the environmental impact), and was inversely proportional to the per-capita usable area of biologically productive land (the greater the EF, the smaller the per-capita usable area of biologically productive land) [49]. It is now a widely used measure in the ﬁeld of ecological economics as it is a quantitative indicator that is easy to understand and calculate. Therefore, this paper uses sustainable ecological evaluation indicators (SEEIs) to measure regional ecological security on a per-unit ecological footprint basis. In summary, this paper ﬁrst combines the ecosystem services and ecological footprint models to evaluate the sustainability status based on each of the ecosystem service features for the different Gaomei wetlands land use covers, after which the ecosystem service values are calculated. 2.2. Methods This study combines ecosystem services and the EF model to reclassify the EF ecosystem according to the various land-use features for each ecosystem service system at the Gaomei wetlands. The ecological footprint (EF) (demand) and ecological capacity (supply) at the Gaomei wetlands were ﬁrst evaluated over various periods using an EF model developed from ecosystem services theory, after which an eco-security indicator system was established to estimate the Gaomei wetlands’ eco-security, the steps for which were as follows: 1. Introduction Subsequently, the SEEI—the ecological remainder (ER), the ecological deﬁcit (ED), the EFI and ESI—is used to analyze the resource utilization efﬁciency and ecological security in the Gaomei wetlands. The problems identiﬁed by the different indicator values are evaluated to develop a systematic measurement apparatus to encourage sustainable development and to review the evolution in sustainable development trends. 2.1. Study Area The Gaomei wetlands (24◦18′35.07′′ N, 120◦33′08.21′′ E) are located in Shimizu, Taichung, Taiwan. The Gaomei wetlands support diverse bird species and special habitats such as lagoons and sandbars, providing many possible tourist opportunities. As the Gaomei wetlands have been evolving in the past few years from a primarily agriculturally based economy to a primarily tourism based economy, there have been several recent studies focused on tourist behavior and local tourism support initiatives [50–52]. 2.2.1. Ecosystem Services Value Model The methods used to estimate the ecosystem service value in the research area were based on the value assessment method models outlined in [10,53,54]. An equivalence factor for the ecosystem service value for the different land uses and land cover at the Gaomei wetlands was calculated using 4 of 11 Water 2017, 9, 197 the assessment model, which took the ecosystem service value for the different land use and land covers as the basis for determining the ecosystem service value, as shown in Equation (1): ESVk = ∑ m i ∑ n j Ai × fij × Ea × Sk × Tk (1) (1) where ESV is the total ecosystem service value; Ai is the distribution area for the ith type of land use and land cover (gha); fij is the equivalence factor for the jth ecosystem goods and services item provided by the ith ecosystem; Ea is the production per unit area or the ecosystem service value coefﬁcient; Sk is a K coefﬁcient for regional differences; Tk is a K regional service support coefﬁcient; i is the land use and land covers in the different ecosystems; j is the ecosystem service category. 2.2.2. Accommodation Ecological Footprint The EF concept [55,56] was used to evaluate the changes in the Gaomei wetlands from 2008 to 2015. To examine the inﬂuence of the EF on the environment, the transportation ecological footprint (TREF), the activities ecological footprint (ACTEF), and the food and ﬁber consumption ecological footprint (FEF) were employed as the evaluation measures. The general formulas for calculating the EF and ECC are shown in Equations (2) and (3): EF = N × ef = N × n ∑ i=1 (a × ai) = N × 6 ∑ j=1 rj n ∑ i=1 ci pi ! (i = 1, 2, 3, . . . n; j = 1, 2, 3, . . . , 6) (2) ECC = N × 6 ∑ j=1 ecj = N × 6 ∑ j=1 Aj × rj × yj , (j = 1, 2, 3, . . . , 6) (3) (2) ECC = N × 6 ∑ j=1 ecj = N × 6 ∑ j=1 Aj × rj × yj , (j = 1, 2, 3, . . . , 6) (3) (3) in which EF is the total EF (gha); N is the total population; ef is the EF per capita (gha); aai is the biologically productive area per capita (gha), converted to the ith traded commodity type; ci is the consumption per capita (kg) of the ith commodity type; pi is the average global productive capacity (kg/(t/gha)) of the ith consumer goods type; rj and yj are the equivalence factor and yield factor (YF) for the jth land type; j is the corresponding land use or land cover type; ECC denotes the total ecological carrying capacity; ecj is the ECC per capita; and Aj is the area per capita of the jth land type in the region. 2.2.3. Ecological Footprint Model The traditional ecological footprint model converts the EF and ECC of the land ecosystems into biological resource consumption (agricultural land, forest land, grassland, and ﬁsheries) and energy consumption (carbon footprint, completion land) and six other ecological system units. However, as this model includes systems and services ecological functions in the ECC calculation, it assumes that the ecosystem services of the various land ecosystem supply units can be used as fossil fuels. Therefore, the ECC evaluation of the ecosystem service value for the land ecosystems is reclassiﬁed in this paper to (1) agricultural ecosystem; (2) forest ecosystem; (3) grassland ecosystem; (4) ecosystem completions; (5) ﬁsheries ecosystems (including lakes, rivers and wetlands); (6) unutilized land; and six other ecosystem units. Traditional ecological footprint calculations consider only the production of food and raw materials to provide the two ecological functions. However, in the integrated EF model, an appropriate equivalence factor and a yield factor (YF) are used to determine the ecological functions for the biological land productivity and food production raw materials rather than the ecological functions and the ecosystem service value. YF is used mainly to reﬂect the differences in the different regions per unit area ecosystem services in the Gaomei wetlands, which is calculated as follows: YFj = vj/v− j (4) (4) YFj = vj/v− j 5 of 11 Water 2017, 9, 197 in which YFj is the yield factor for the jth type of ecosystem unit, j = 1, 2, . . . , 6, are the 6 ecosystem units; vj is the per unit area value of the ecosystem service function for the jth type of ecosystem in a region; and v− j is the per unit area ecosystem service function value for the jth type of ecosystem in Taiwan. Using Equation (4), the Gaomei wetlands yield factor was calculated from 2008 to 2015, as shown able 1. Using Equation (4), the Gaomei wetlands yield factor was calculated from 2008 to 2015, as shown in Table 1. Using Equation (4), the Gaomei wetlands yield factor was calculated from 2008 to 2015, as shown in Table 1. Table 1. Yield factor (YF) for different land use and land cover ecosystems in the Gaomei wetlands from 2008 to 2015. Table 1. Yield factor (YF) for different land use and land cover ecosystems in the Gaomei wetlands from 2008 to 2015. 2.2.3. Ecological Footprint Model Year Agricultural Ecosystem Forest Ecosystem Grassland Ecology System Fisheries Ecosystems Ecosystem Completions Unutilized Land and Six Other Ecosystem Units 2008 1.51 0.92 1.15 1.88 0.62 1.92 2009 1.49 0.91 1.13 1.81 0.58 2.01 2010 1.48 0.88 1.12 1.80 0.54 2.15 2011 1.31 0.77 1.14 1.60 0.40 1.98 2012 1.06 0.60 0.87 1.28 0.15 1.62 2013 1.14 0.63 0.94 1.33 0.15 1.77 2014 1.07 0.57 0.88 1.21 0.11 1.64 2015 1.00 0.52 0.77 1.11 0.08 1.56 2.2.4. Sustainable Ecological Evaluation Indicators (SEEI) 2.2.4. Sustainable Ecological Evaluation Indicators (SEEI) Multiple quantitative indicators (e.g., ED, ER, EFI and ESI) were employed to develop an indicator set to evaluate the Gaomei wetlands ecological sustainability and to establish the standards so as to be able to properly assess the ecological security. The evaluation indicators utilized in this research are outlined in the following subsections. (a) ED or ER When the EF is lower than the ECC of a region, there is an ecological remainder (ER), indicating that the corresponding development model is sustainable. When the EF is higher than the ECC of a region, there is an ecological deﬁcit (ED), indicating that the corresponding development model is not sustainable. As the ecological deﬁcit results from excessive human resource demands [57], demand must be reduced to achieve sustainable ecological development. The formulas for ED and ER are shown as Equations (5) and (6). ER = ECC −EF (5) ED = EF −ECC (6) (5) (5) (6) (5) (6) in which ER is the ecological remainder, ED is the ecological deﬁcit, ECC is the ecological carrying capacity, and EF is the ecological footprint. in which ER is the ecological remainder, ED is the ecological deﬁcit, ECC is the ecological carrying capacity, and EF is the ecological footprint. (b) EFI (c) ESI The Environmental Sustainability Index (ESI), which was developed by the Center for International Earth Science Information Network (CIESIN), the Yale Center for Environmental Law and Policy (YCELP), and the World Economic Forum, assesses sustainability by measuring the degree to which the ecological systems in a region meet the human ecological demands. The ESI formula is shown in Equation (8) and the ESI levels are given in Table 3 ESI = ECC/(ECC + EF) (8) (8) in which ESI is the environmental sustainability index, EF is the ecological footprint; and ECC is the ecological carrying capacity. in which ESI is the environmental sustainability index, EF is the ecological footprint; and ECC is the ecological carrying capacity. Table 3. Environmental Sustainability Index (ESI) levels. Level ESI Regional Ecological Sustainability Extent 1 >0.7 High sustainability 2 0.50~0.70 Low sustainability 3 0.30~0.50 Low unsustainability 4 <0.30 High unsustainability Resource: [58]. Table 3. Environmental Sustainability Index (ESI) levels. Table 3. Environmental Sustainability Index (ESI) levels. Table 3. Environmental Sustainability Index (ESI) levels. Table 2. Ecological Footprint Index (EFI) levels and conditions. Level EFI EFI Conditions 1 <0.5 Safe 2 0.5~0.8 Moderately safe 3 0.8~1.0 Threshold 4 >1.0 Unsafe Resource: [49]. Level EFI EFI Conditions 1 <0.5 Safe 2 0.5~0.8 Moderately safe 3 0.8~1.0 Threshold 4 >1.0 Unsafe Resource: [49]. (c) ESI (b) EFI The EFI compares the resources and energy expenditures to the region’s ECC to assess resource utilization and determine development sustainability. The EFI formula is as in Equation (7), and the EFI levels are as shown in Table 2 EFI = EF/ECC (7) EFI = EF/ECC (7) in which EFI is the ecological footprint index, EF is the ecological footprint, and ECC is the ecological carrying capacity. 6 of 11 Water 2017, 9, 197 3.1. Ecosystem Services Value Computation and Analysis Results 3.1. Ecosystem Services Value Computation and Analysis Results 3.1. Ecosystem Services Value Computation and Analysis Results Equation (2) was used to calculate the ecological service value of each ecosystem type in the Gaomei wetlands from 2008 to 2015, as shown in Table 4. The total value of the ecosystem services in the Gaomei wetlands increased from 59.24 million TWD to 98.10 million TWD over the 8 years. From 2008 to 2013, the total Gaomei wetlands ecosystem service value increased by 42.27 million TWD due to the increased ﬁsheries (including lakes, rivers, and wetlands) and agricultural land areas, both of which had large ecological service value coefﬁcients. The total Gaomei wetlands ecosystem service value increased from 2013 to 2015; however, during the same period, the Gaomei wetlands total ecosystem service value decreased by 3.41 million TWD because the ﬁsheries area, which had a large ecological service value coefﬁcient decreased rapidly and the increased agricultural land area was not sufﬁcient to compensate for the ﬁsheries reduced ecological service value. Since 2013, therefore, the Gaomei wetlands total ecosystem service value has been decreasing. From 2008 to 2015, the relative value of each Gaomei wetlands ecosystem type (Table 4) changed variably. The ﬁsheries ecosystem services function value increased then decreased, the forest ecosystem services function value increased, decreased, and then increased, and the grassland ecosystem services function value had an M-shaped pattern: increase–decrease–increase–decrease; the agricultural ecosystem services function value decreased and then increased, with the overall ecosystem services function value increasing. Taken together, the total ecosystem service value in increased by a net of 65.60%, with the ﬁsheries ecosystem service value increasing the most (81.09%), and that of the forest ecosystems decreasing the most (−84.38%) followed by the grassland ecosystems 7 of 11 Water 2017, 9, 197 (−33.39%). The ﬁsheries accounted for 39% of the total utilized land area in the study area; however, the ﬁsheries ecological service value accounted for 75.56% of the total ecosystem service value. Because the ﬁsheries in this area have a high ecosystem service value coefﬁcient, this land use type has a higher total ecosystem service value, highlighting the importance of the ﬁsheries in the Gaomei wetlands ecosystem. Table 4. Ecosystem service values from 2008 to 2015 for each Gaomei wetlands ecosystem type. 3.1. Ecosystem Services Value Computation and Analysis Results Year Fisheries Ecosystems Grassland Ecology System Forest Ecosystem Agricultural Ecosystem The Completion of Ecosystems Unutilized land and Six Other Ecosystem Units Total Ecosystem Services Values (106 TWD) 2008 40.93 6.17 0.32 10.68 0.01 1.13 59.24 2009 48.85 6.28 0.34 10.74 0.01 1.27 67.49 2010 53.12 6.35 0.34 10.82 0.01 1.35 71.99 2011 60.89 6.39 0.36 10.10 0.01 1.44 79.19 2012 65.11 6.62 0.55 12.08 0.01 1.39 85.76 2013 83.79 3.34 0.04 12.76 0.01 1.57 101.51 2014 78.18 4.37 0.05 14.56 0.01 1.44 98.61 2015 74.12 4.11 0.05 18.49 0.02 1.31 98.10 3.2. EF Computation and Analysis Results Table 4. Ecosystem service values from 2008 to 2015 for each Gaomei wetlands ecosystem type. 3.2. EF Computation and Analysis Results 3.2. EF Computation and Analysis Results Table 5 details the three Gaomei wetlands activity types and the EF computations. The EF gradually increased from 244.03 global hectares (gha) in 2008 to 380.98 gha in 2015. Of the three EF activity types, the TREF had the largest proportion at an average of 70.18%, followed by the ACTEF at an average of 24.97% and the FEF at an average of 4.85%. Based on these empirical results, the TREF grew from 140.47 gha in 2008 to 267.36 gha in 2015 due to a signiﬁcant growth in tourist numbers, which caused an increased demand for vehicles, further inﬂating liqueﬁed fuel demands. Table 5. Ecological Footprint (EF) for the three activities types and total Ecological Footprint (EF) (unit: gh ble 5. Ecological Footprint (EF) for the three activities types and total Ecological Footprint (EF) (unit: gha). Table 5. Ecological Footprint (EF) for the three activities types and total Ecological Footprint (EF) (unit: gha). Year TREF ACTEF FEF EF 2008 140.47 95.14 8.42 244.03 2009 183.70 95.14 13.52 292.36 2010 204.87 95.14 13.37 313.38 2011 184.57 95.14 13.77 293.48 2012 182.99 95.14 12.30 290.43 2013 220.37 95.14 15.54 331.05 2014 267.04 95.14 19.96 382.14 2015 267.36 95.14 18.48 380.98 Average proportion 70.18% 24.97% 4.85% 100.000% Notes: TREF: transport ecological footprint; ACTEF: activities footprint; FEF: food & ﬁber consumption ecological footprint; EF: total ecological footprint. As the Gaomei wetlands gross area did not change substantially between 2008 and 2015, the ACTEF was steady at 95.14 gha; however, the FEF increased from 8.42 gha in 2008 to 18.48 gha in 2015. 3.1. Ecosystem Services Value Computation and Analysis Results The FEF indicates the tourist dietary demands inside the Gaomei wetlands and includes grains, coarse cereals, vegetables, fruit, meat, and ﬁsh; therefore, the substantial growth in the FEF was linked to the growing tourist numbers. This analysis found that the main tourism ecological resource consumption came from the fossil energy used by the vehicles traveling between residences and destinations and the increase in the land accessible to tourists for leisure activities. 3.3. SEEI Computation and Analysis Results The SEEI indicated that the ED grew by about 106.54% from 2008 to 2015. In 2015, the EFI was rated Level 4 at 1.02, and the ESI was rated Level 3 at 0.49, indicating that the Gaomei wetlands were at an unsafe ecological security level within that time span. 8 of 11 Water 2017, 9, 197 (c) ESI Table 6 shows the computational results for the ESI. Between 2008 and 2013, the ESI remained at Level 2, signifying low sustainability. Yet from 2014 onward, the ESI has fallen to Level 3, signifying unsustainability. If this situation is not controlled and improved, the attainment of sustainable ecological development will be impossible. Table 6. Ecological Footprint (EF), ecological carrying capacity (ECC), ecological remainder (ER)/ecological deﬁcit (ED), ecological footprint index (EFI), and environmental sustainability index (ESI) (unit: gha). Year ECC EF ES/ED EFI ESI Index Level Representation Condition Index Level Representational State 2008 372.57 244.03 128.54 0.65 2 Moderately safe 0.60 2 Low sustainability 2009 372.57 292.36 80.18 0.78 2 Moderately safe 0.56 2 Low sustainability 2010 372.57 313.38 59.19 0.84 3 Threshold 0.54 2 Low sustainability 2011 372.57 293.48 79.09 0.79 2 Moderately safe 0.56 2 Low sustainability 2012 372.57 290.43 82.14 0.78 2 Moderately safe 0.56 2 Low sustainability 2013 372.57 331.05 41.52 0.89 3 Threshold 0.53 2 Low sustainability 2014 372.57 382.14 −9.57 1.03 4 Unsafe 0.49 3 Low unsustainability 2015 372.57 380.98 −8.41 1.02 4 Unsafe 0.49 3 Low unsustainability Table 6. Ecological Footprint (EF), ecological carrying capacity (ECC), ecological remainder (ER)/ecological deﬁcit (ED), ecological footprint index (EFI), and environmental sustainability index (ESI) (unit: gha). (a) ER/ED Table 6 gives the computation results for the ER/ED. The ER had a declining trend from 128.54 gha in 2008 to 41.52 gha in 2013. The Gaomei wetlands ecological deﬁcit (ED) decreased from −9.57 gha in 2014 to −8.41 gha in 2015, primarily due to the increased tourist numbers. (b) EFI This study utilized the EFI to evaluate the Gaomei wetlands ecological security, the outcomes of which are shown in Table 6. The EFI increased from 0.65 in 2008 to 1.02 in 2015, indicating an unsafe ecological security level, which is predicted to further increase because of the rising tourist demand for resources and services (e.g., bus services, road construction, and waste production). ( ) ESI References 1. 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Global positioning systems could be used in rental cars to monitor tourist activity, which can then be used to develop effective transportation systems aimed at decreasing overall fossil fuel use and minimizing the associated carbon footprints. Acknowledgments: I am thankful to the Ministry of Science and Technology (Republic of China, Taiwan) for ﬁnancially supporting this research project (grant number MOST 102-2410-H-040-010). Conﬂicts of Interest: The authors declare no conﬂict of interest. Conﬂicts of Interest: The authors declare no conﬂict of interest. 4. Conclusions This study employed the ecosystem service value, ecological capacity, EF, and sustainable ecological evaluation indicators (SEEIs) to assess the ecological security and the efﬁcient use of resources in the Gaomei wetlands. We came to the following conclusions: The total value of the ecosystem services in the Gaomei wetlands increased from 59.24 million TWD in 2008 to 98.10 million TWD in 2015. The EF gradually increased from 244.03 gha in 2008 to 380.98 gha in 2015. Of the three activity EFs, TREF had the biggest proportion (70.18%), with ACTEF (24.97%) and FEF (4.85%) following thereafter. The SEEI indicated that the ED grew by about 106.54% from 2008 to 2015. In 2015, the EFI was rated Level 4 at 1.02, and the ESI was rated Level 3 at 0.49. Therefore, as the Gaomei wetlands are predicted to become ecologically unsustainable over time, local, regional, and national governments need to implement regulations to strictly control the Gaomei wetlands land use. According to the empirical analysis results, the primary factors inﬂuencing various types of activity EFs are presented below. (a) Tourists had a negative effect on the overall EF from all activities. Therefore, when tourist numbers increased, the EF increased and there was a greater environmental impact. 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This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). © 2017 by the author. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
https://openalex.org/W4226224638	http://journal.unisa-bandung.ac.id/index.php/jka/article/download/284/156	Indonesian	null	HUBUNGAN PENGAWAS MENELAN OBAT (PMO) DENGAN KEPATUHAN MINUM OBAT PENDERITA TUBERKULOSIS	Jurnal Keperawatan 'Aisyiyah	2,021	cc-by	3,169	JKA.2021;8(2):133-139 JKA.2021;8(2):133-139 ARTIKEL PENELITIAN HUBUNGAN PENGAWAS MENELAN OBAT (PMO) DENGAN KEPATUHAN MINUM OBAT PENDERITA TUBERKULOSIS Yayat Hidayat¹, Hendara Gunawan² Universitas ‘Aisyiyah Bandung yayat.hidayat@unisa-bandung.ac.id RELATIONSHIP OF SWALLOWING DRUG SUPERVISORS WITH THE COMPLIANCE WITH THE DRUG OF TUBERCULOSIS PATIENTS RELATIONSHIP OF SWALLOWING DRUG SUPERVISORS WITH THE COMPLIANCE WITH THE DRUG OF TUBERCULOSIS PATIENTS ABSTRAK Penanggulangan Tuberkulosis oleh ‘Aisyiyah dilakukan mengingat kasus Tuberkulosis di Kabupaten Bandung termasuk daerah tertinggi di Jawa Barat mencapai 2.362 kasus. sedangkan kasus Tuberkulosis Multiple Drug Resistance (MDR) mencapai 35 kasus pada tahun 2013. Kegiatan yang dilaksanakan oleh TB Care ‘Aisyiyah antara lain berupa kegiatan pelatihan bagi Pengawas Menelan Obat (PMO), Kegiatan tersebut meliputi tugas PMO dalam mengawasi minum obat, mendampingi penderita dan memotivasi penderita TB agar minum obat secara rutin sampai sembuh.Tujuan dari penelitian ini untuk menganalisis hubungan PMO dengan kepatuhan minum obat penderita TB di Kabupaten Bandung. Lokasi penelitian di 5 tempat wilayah kerja puskesmas Cileunyi, Rancaekek, Cicalengka Kabupaten Bandung. Target penelitian ini adalah ditemukannya kepatuhan minum obat penderita TB yang mempunyai PMO hasil pelatihan organisasi ‘Aisyiyah Kabupaten Bandung. Penelitian ini menggunakan desain cross-sectional menghubungkan antara faktor PMO dengan tingkat kepatuhan minum obat penderita TB. Populasi pada penelitian ini seluruh pasien penderita TB paru yang berobat ke Puskesmas Rancaekek, Cicalengka, Cileunyi, berjumlah 205 orang adapun jumlah sampel 124 orang. Pengumpulan data dilakukan pada bulan Desember 2016 sampai dengan Maret tahun 2017. Hasil penelitian ini ada hubungan signifikan antara PMO dengan Kepatuhan minum obat penderita TB. Kata kunci: ‘Aisyiyah, kepatuhan, pengawas menelan obat PENDAHULUAN diawasi langsung oleh Pengawas Menelan Obat (PMO),(Kemenkes, 2011) Terdapat 5 komponen pada strategi DOTS antara lain komitmen politis, pemeriksaan dahak mikroskopis yang terjamin mutunya, pengobatan jangka pendek yang standar bagi semua kasus TB dengan tata laksana kasus yang tepat termasuk pengawasan langsung pengobatan, jaminan ketersediaan OAT yang bermutu serta sistem pencatatan dan pelaporan yang mampu memberikan penilaian terhadap hasil pengobatan penderita dan kinerja program secara keseluruhan. Pada tahun 2014 incidence absolute number dari 20 negara tertinggi TB, India berada pada urutan pertama dengan angka kejadian 22,7%, Indonesia berada pada urutan kedua dengan angka kejadian 10,3%, China berada pada urutan ketiga dengan angka kejadian 9,6%, Nigeria berada pada urutan ke empat dengan angka kejadian 5,9%, dan Pakistan berada pada urutan kelima dengan angka kejadian 5,2% dari total dunia (Kemenkes, 2012). Penyakit Tuberkulosis (TB) merupakan penyakit menular yang menjadi masalah kesehatan di Indonesia termasuk di Jawa Barat. Berdasarkan data dari profil kesehatan Indonesia tahun 2013, Provinsi Jawa Barat merupakan daerah yang mempunyai prevalensi TB paru tertinggi di Indonesia, yaitu sebesar 0,7% (Kesehatan, 2014). Kasus Basil Tahan Asam (BTA) positif di Jawa Barat pada tahun 2013 sejumlah 33.460. Di Jawa Barat terdapat lima Kabupaten/Kota yang mempunyai Kasus TB tertinggi, salah satunya adalah Kabupaten Bandung.(Jabar, 2013). Berdasarkan data dari Dinas Kesehatan Kabupaten Bandung, kasus BTA positif di Kabupaten Bandung pada tahun 2013 sejumlah 2.362 serta terdapat 35 orang yang termasuk kategori TB MDR (Multiple Drug Resistance)/penderita TB yang kebal terhadap obat anti Tuberkulosis (OAT) (Riantini, 2014). Analisis kinerja PMO dengan kesembuhan TB, bahwa 23 orang (100%) penderita TB yang sembuh memiliki kinerja PMO yang baik. Pasien TB dengan kinerja PMO yang baik lebih besar kemungkinan untuk sembuh. Keberhasilan pengobatan tuberkulosis paru ditentukan oleh kepatuhan dan keteraturan dalam berobat, pemeriksaan fisik, dan laboratorium. Oleh sebab itu, selama masa pengobatan diperlukan kerja sama yang baik serta berkesinambungan antara PMO dengan penderita dalam mematuhi peraturan tata cara minum obat dan kontrol kesehatan.(Ariani et al., 2015). Analisis kinerja PMO dengan kesembuhan TB, bahwa 23 orang (100%) penderita TB yang sembuh memiliki kinerja PMO yang baik. Pasien TB dengan kinerja PMO yang baik lebih besar kemungkinan untuk sembuh (Hayati, 2016). Penelitian (Yoisangadji, 2016) menyimpulkan bahwa terdapat hubungan anatara PMO dengan kepatuhan minum obat TB pada pasien TB di wilayah kerja Puskesmas Sario. Upaya untuk memberantas penyakit Tuberkulosis (TB) dilakukan diantaranya dengan menggunakan strategi DOTS (Directly Observed Treatment Shortcourse) yang direkomendasikan oleh World Health Organization (WHO) (Kemenkes RI, 2012). Abstract Tuberculosis control by ‘Aisyiyah carries out considering that Tuberculosis cases in Bandung Regency were among the highest areas in West Java, reaching 2,362 cases. At the same time, the Tuberculosis Multiple Drug Resistance (MDR) cases reached 35 cases in 2013. Activities carried out by TB Care ‘Aisyiyah include training activities for Drug Swallowing Supervisors (PMO). Motivate TB patients to take medication regularly until they recover. This study aimed to analyze the relationship between PMO and adherence to medication for TB patients in Bandung Regency. The research locations are in 5 working areas of the Cileunyi Public Health Center, Rancaekek, Cicalengka, Bandung Regency. This research aims to find adherence to medication for TB patients who have PMO due to the training of the organization ‘Aisyiyah, Bandung Regency. This study used a cross- sectional design linking the PMO factor with the level of adherence to medication for TB patients. The population in this study were all patients with pulmonary TB who treat at the Rancaekek Health Center, Cicalengka, Cileunyi, which amounted to 205 people. In comparison, the number of samples was 124 people. Data collection carries out from December 2016 to March 2017. This study showed a significant relationship between PMO and adherence to medication for TB patients. Keywords: ‘Aisyiyah, compliance, drug swallowing supervisor 133 Jurnal Keperawatan ‘Aisyiyah 134 PENDAHULUAN Upaya untuk menanggulangi penyakit TB di Indonesia dilakukan secara terpadu melalui Gerakan Terpadu Nasional Tuberkulosis (Gerdunas). Upaya ini melibatkan berbagai pihak terkait, yang salah satunya adalah dilibatkannya organisasi ‘Aisyiyah. Organisasi ‘Aisyiyah merupakan organisasi yang bertanggungjawab Ada lima komponen dalam strategi DOTS, yaitu Komitmen politis dari pemerintah untuk menjalankan program TB nasional, diagnosis TB melalui pemeriksaan dahak secara mikroskopis, Pengobatan TB dengan paduan Obat Anti Tuberkulosis (OAT) yang JKA \| Volume 8 \| Nomor 2 \| Desember 2021 Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan Minum Obat Penderita Tuberkulosis 135 135 Tantangan yang dihadapi oleh PMO khususnya di wilayah kerja TB ‘Aisyiyah Kabupaten Bandung semakin berat. Hal ini berdasarkan data dari Dinas Kesehatan Kabupaten Bandung per tri wulan II tahun 2015, Kecamatan Cileunyi dan Rancaekek merupakan daerah tertinggi cakupan penemuan kasus baru BTA positif. Selain itu, kasus TB di Kabupaten Bandung semakin kompleks dengan meningkatnya jumlah penderita TB MDR (Multiple Drug Resistance) yang kebal terhadap OAT biasa. Jumlah Penderita TB MDR per triwulan II tahun 2015 sebanyak 58 orang yang meningkat dari tahun 2013 sejumlah 18 orang dan tahun 2014 sejumlah 41 orang. Dari aspek finansial, jumlah dana yang harus dikeluarkan oleh pemerintah daerah untuk mengobati satu orang penderita TB MDR lebih kurang sejumlah 120 juta rupiah, sehingga apabila semakin banyak penderita TB MDR maka akan menghabiskan dana milyaran rupiah untuk pengobatan penderita TB MDR tersebut. untuk melaksanakan program Community TB Care, yaitu penanggulangan TB yang berbasis komunitas yang melibatkan seluruh masyarakat sehingga menumbuhkan kemandirian masyarakat dalam mengendalikan penyakit TB (Wai et al., 2018). Program Community TB Care ‘Aisyiyah bertujuan menggerakkan masyarakat untuk peduli terhadap penyakit TB dan dapat menggalang seluruh komponen masyarakat untuk berupaya memberantas penyakit TB. Kegiatan yang dilaksanakan adalah pemberdayaan anggota masyarakat diantaranya melalui kegiatan pelatihan Pengawas Menelan Obat (PMO). PMO merupakan salah satu kegiatan ‘Aisyiyah yang bertujuan menghasilkan anggota masyarakat yang mampu secara mandiri untuk mengawasi minum obat bagi penderita TB (Zulfikar, 2015). Penanggulangan TB oleh organisasi ‘Aisyiyah dilaksanakan di berbagai Provinsi dan Kabupaten/Kota di Indonesia termasuk di Kabupaten Bandung. Pimpinan Daerah ‘Aisyiyah (PDA) Kabupaten Bandung mendapat kepercayaan dari Pimpinan Pusat ‘Aisyiyah (PPA) untuk melaksanakan program TB di wilayah Kabupaten Bandung. Program TB yang dilaksanakan oleh PDA Kabupaten Bandung dimulai sejak bulan Januari 2014 yang dilaksanakan di wilayah kecamatan Cileunyi, Rancaekek, Cicalengka, Katapang dan Margahayu. JKA \| Volume 8 \| Nomor 2 \| Desember 2021 PENDAHULUAN Oleh karena itu, PMO merupakan salah satu upaya untuk mendampingi pengobatan penderita TB supaya penyakitnya sembuh, tidak menular ke orang lain dan dapat mencegah menjadi penderita TB MDR bahkan TB XDR (Extra Drug Resistance). Pelatihan PMO yang dilaksanakan oleh ‘Aisyiyah Kabupaten Bandung sudah dilaksanakan sejak bulan September 2014 di 5 Kecamatan, yaitu Cicalengka, Cileunyi, Rancaekek, Katapang dan Margahayu dengan jumlah PMO terlatih sebanyak 180 orang, akan tetapi hubungan PMO dengan kepatuhan minum obat penderita TB di 5 lokasi tersebut sampai saat ini belum pernah dilakukan penelitian. Program TB yang telah dilaksanakan oleh PDA Kabupaten Bandung antara lain berupa kegiatan pelatihan TB bagi PMO yang berasal dari keluarga penderita TB. Tugas kader yang menjadi PMO, yaitu melakukan pengawasan dan pendampingan minum obat bagi penderita TB yang sedang menjalani pengobatan, mengantar penderita TB memeriksakan dahak dan berobat ke unit pelayanan kesehatan (UPK) serta memastikan bahwa penderita yang didampingi pengobatannya sampai selesai dan dinyatakan sembuh oleh tenaga kesehatan. Tujuan Penelitian untuk menganalisis hubungan Pengawas Menelan Obat (PMO) dengan kepatuhan minum obat penderita TB di Kabupaten Bandung. Manfaat penelitian sebagai masukan pentingnya meningkatkan partisipasi, kemandirian dan pemberdayaan masyarakat JKA \| Volume 8 \| Nomor 2 \| Desember 2021 Jurnal Keperawatan ‘Aisyiyah 136 khususnya kader PMO dalam ikut menanggulangi penyakit TB. bawah ini. bawah ini. Tabel 1. Distribusi frekuensi (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung Kategori n % Tidak ada PMO 62 50,0 Ada PMO 62 50,0 Total 124 100 Tabel 1. Distribusi frekuensi (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung Tabel 1. Distribusi frekuensi (PMO) di METODOLOGI Penelitian ini menggunakan desain case control yang menghubungkan antara faktor PMO dengan tingkat kepatuhan minum obat penderita TB, yang artinya setiap subyek penelitian diobservasi hanya satu kali saja/ dan faktor risiko serta dampak diukur menurut keadaan atau status pada saat observasi. Populasi pada penelitian ini adalah seluruh penderita penderita TB paru yang berobat ke Tabel di atas memberikan gambaran bahwa jumlah responden tidak ada PMO sama banyaknya dengan jumlah responden yang ada PMO di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Puskesmas Rancaekek, Cicalengka, Cileunyi, Margahayu dan Katapang berjumlah 205 orang serta PMO yang telah dilatih oleh ‘Aisyiyah Kabupaten Bandung di 5 lokasi penelitian. Dengan jumlah sampel 62 responden. Pengumpulan data dilakukan pada bulan September tahun 2016 sampai dengan Juni tahun 2017 di Puskesmas Rancaekek, Cicalengka, Cileunyi, Margahayu dan Katapang Kabupaten Bandung. Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Hasil pengolahan data terhadap variabel Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung dapat digambarkan seperti tabel di bawah ini. Tabel 2. Distribusi frekuensi Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung Kategori n % Tinggi 85 68,5 Sedang 18 13,7 Rendah 21 17,7 Total 124 100 HASIL Tabel 2. Distribusi frekuensi Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Tabel 2. Distribusi frekuensi Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung Penelitian ini dilaksanakan di wilayah kerja Puskesmas Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung pada tanggal 26 Desember tahun 2016 sampai dengan tanggal 18 Maret 2017. Pengolahan data dilakukan pada tanggal 3 April sampai 8 April 2017. Hasil penelitian ini disajikan melalui dua tahap analisis, yaitu analisis univariat dan bivariat yang selengkapnya diuraikan seperti di bawah ini. Rancaekek, Cicalengka, Kabupaten Bandung Kategori n % Tinggi 85 68,5 Sedang 18 13,7 Rendah 21 17,7 Total 124 100 Tabel di atas memberikan gambaran bahwa responden yang memiliki kepatuhan yang tinggi minum OAT sebesar 68,5%, kepatuhan sedang 13,7%, dan kepatuhan rendah 17,7% untuk minum obat pada penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Gambaran Pengawas Menelan Obat (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Hasil pengolahan data terhadap variabel Pengawas Menelan Obat (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung dapat digambarkan seperti tabel di JKA \| Volume 8 \| Nomor 2 \| Desember 2021 Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan Minum Obat Penderita Tuberkulosis 137 137 Tabel 3. Analisis Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Pengawas Menelan Obat Kepatuhan chi- square p-value Tinggi % Sedang % Rendah % Total % Ada 48 38,7 10 8,1 4 3,2 62 50,0 8,028 0,018 Tidak Ada 37 29,8 8 6,5 17 13,7 62 50,0 Total 85 68,5 18 14,5 21 16,9 124 100 Tabel 3. Analisis Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Tabel 3. Analisis Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Merujuk pada tabel di atas dapat diinformasikan bahwa penderita TB yang ada PMO memiliki kepatuhan yang tinggi lebih besar persentasenya (38,7%) dibandingkan penderita TB yang tidak ada PMO (29,8%) walaupun jumlah mereka sama banyaknya, dan pada tingkat kepatuhan rendah persentase penderita TB yang tidak ada PMO (13,7%) lebih besar dibanding yang ada PMO (3,2%). Analisis chi Square diperoleh nilai probabilitas (p-value) 0,018 lebih kecil dari alpa 0,05. Hal ini berarti ada hubungan signifikan antara Pengawas Menelan Obat (PMO) dengan kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. HASIL Keberadaan PMO adalah penting setidaknya selama tahap pengobatan intensif (2 bulan pertama) untuk meyakinkan bahwa obat dimakan dengan kombinasi yang benar serta sesuai dengan jadwal yang ditentukan. Melalui pengawasan pengobatan secara langsung, pasien dapat dipantau tingkat kepatuhan menelan obat sehingga pengobatan dapat dilakukan secara tuntas sesuai dengan program pengobatan yang telah dijadwalkan. Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Kepatuhan minum obat merupakan kepatuhan pasien TB Paru untuk mematuhi prosedur pengobatan dan melakukan minum obat sesuai jadwal yang ditentukan oleh Puskesmas atau Tenaga kesehatan. Pengobatan pasien TB dilakukan dengan pemberian obat TB dengan komposisi 2HRZ/4H2R2: INH+Rifampisin+Pirazinamid: setiap hari selama 2 bulan pertama, kemudian INH+Rifampisin setiap hari atau 2 kali seminggu selama 4 bulan (ditambahkan Etambutol bila diduga ada resistensi terhadap INH). Jika tidak dilakukan sesuai jadwal maka ada risiko pasien untuk mengulang pengobatan bahkan resisten terhadap OAT. JKA \| Volume 8 \| Nomor 2 \| Desember 2021 SIMPULAN DAN SARAN Dari hasil analisis chi-square diperoleh nilai probabilitas (p-value) 0,018 lebih kecil dari alpa 0,05. Hal ini berarti ada hubungan signifikan antara Pengawas Menelan Obat (PMO) dengan Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Bagi organisasi ‘Aisyiyah perlunya menjalin kerjasama dengan pihak lain dalam penanggulangan penyakit TB yang dilakukan secara sabilulungan sehingga penanggulangan penyakit TB dapat secara komprehensif dan melibatkan banyak pihak.Bagi pemerintah daerah Kabupaten Bandung khususnya Dinas Kesehatan dan aparat kewilayahan kecamatan dan desa hendaknya membuat program pemberdayaan bagi mantan penderita TB sehingga dapat hidup lebih produktif secara ekonomi dan dapat diterima di lingkungan sosialnya dengan baik. Berdasarkan hasil penelitian ini terdapat hubungan yang positif dan signifikan antara peran Pengawas Menelan Obat (PMO) terhadap kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Pada aspek kepatuhan minum obat, walaupun jumlah responden sama, pada tingkat kepatuhan pasien yang memiliki PMO mempunyai persentase lebih tinggi dibandingan dengan yang tidak ada PMO, dengan perbandingan 38,7% berbanding 29,8%. Sedangkan pada tingkat kepatuhan rendah persentase penderita TB yang tidak ada PMO sebesar 13,7% lebih besar dibandingkan yang ada PMO yang sebesar 3,2%. PEMBAHASAN Gambaran Pengawas Menelan Obat (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Berdasarkan hasil pengolahan data terhadap variabel Pengawas Menelan Obat (PMO) di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. Jumlah responden yang dijadikan penelitian totalnya berjumlah 124 responden. Jumlah tersebut di bagi dua kelompok, 62 orang responden ada PMO dan 62 orang tidak ada PMO. Peran Pengawas Minum Obat (PMO) adalah peran pengawasan yang dilakukan oleh keluarga, masyarakat dan atau tenaga kesehatan dalam hal kepatuhan minum obat, berkunjung ke fasilitas pelayanan kesehatan dan konsultasi ke Hasil penelitian ini menunjukkan bahwa kepatuhan pasien minum OAT bervariasi. Pada tingkat kepatuhan yang tinggi, pasien yang memiliki PMO mempunyai persentase 38,7% JKA \| Volume 8 \| Nomor 2 \| Desember 2021 Jurnal Keperawatan ‘Aisyiyah 138 dibandingkan penderita TB yang tidak ada PMO sebesar 29,8% pada aspek kepatuhan minum obat, walaupun jumlah mereka sama banyaknya. Pada tingkat kepatuhan rendah persentase penderita TB yang tidak ada PMO sebesar 13,7% lebih besar dibandingkan yang ada PMO yang sebesar 3,2%. Hasil penelitian ini sejalan dengan penelitian yang dilakukan (Rahmi et al., 2017). Kepatuhan berobat penderita TB paru dipengaruhi oleh perilaku kesehatan (tingkat ilmu pengetahuan, sikap, dan peran petugas kesehatan) dan peran dari PMO. Peran PMO merupakan Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan minum obat penderita TB di Kecamatan Cileunyi, Rancaekek, Cicalengka, Kabupaten Bandung. dilakukan oleh (Yoisangadji, 2016) terdapat hubungan anatara PMO dengan kepatuhan minum obat TB pada pasien TB di wilayah kerja Puskesmas Sario. Kemudian penelitian yang dilakukan oleh (Dewanty et al., 2016) dihasilkan yang paling banyak PMO buruk responden tidak patuh sebanyak 7 orang (63,63%). Terlihat bahwa responden dengan PMO baik semuanya patuh, yaitu sebanyak 3 orang (27,28%). DAFTAR PUSTAKA Hasil penelitian ini sejalan dengan hasil penelitian lain yang dilakukan oleh Prabowo (2014) di Puskesmas Nogosari Boyolali dengan kesimpulan bahwa PMO berperan terhadap kepatuhan kunjungan penderita TB, yaitu sebesar 81,8% penderita TB mempunyai kunjungan berobat ke Puskesmas Nogosari Boyolali yang termasuk kategori patuh, sedangkan 18,2% mempunyai kunjungan yang termasuk kategori tidak patuh. (Prabowo & Wachidah Yuniartika, 2014). Sejalan juga dengan penelitian lain yang Ariani, N. W., Rattu, A. J. M., & Ratag, B. (2015). Faktor-faktor yang berhubungan dengan keteraturan minum obat penderita tuberkulosis paru di wilayah kerja puskesmas Modayag, kabupaten Bolaang Mongondow Timur. JIKMU, 5(2). Dewanty, L. I., Haryanti, T., & Kurniawan, T. P. (2016). Kepatuhan Berobat Penderita Tb Paru Di Puskesmas Nguntoronadi I Kabupaten Wonogiri. Jurnal Kesehatan, 9(1), 39. https://doi.org/10.23917/jurkes. JKA \| Volume 8 \| Nomor 2 \| Desember 2021 Hubungan Pengawas Menelan Obat (PMO) dengan Kepatuhan Minum Obat Penderita Tuberkulosis 139 v9i1.3406 v9i1.3406 Rahmi, N., Medison, I., & Suryadi, I. (2017). Hubungan Tingkat Kepatuhan Penderita Tuberkulosis Paru dengan Perilaku Kesehatan, Efek Samping OAT dan Peran PMO pada Pengobatan Fase Intensif di Puskesmas Seberang Padang September 2012 - Januari 2013. Jurnal Kesehatan Andalas, 6(2), 345. https:// doi.org/10.25077/jka.v6i2.702 Hayati, D. (2016). Jurnal Ilmu Keperawatan, Vol. IV No. 1 April 2016. Jurnal Ilmu Keperawatan, Vol. IV No. 1, IV(1), 10–18. Jabar, D. kesehatan. (2013). Profil Kesehatan Jawa Barat tahun 2013. Kemenkes. (2011). Pedoman Nasional Pengendalian Tuberkulosis. Kemenkes. (2012). Guidance for Return to Learn After a Concussion. In Pedoman: Vol. (Issue). WHO. 2015. Global Tuberculosis Report. World Health Organization. 20thedition Riantini. (2014). Analisa Situasi TB Kabupaten Bandung. Makalah disampaikan pada kegiatan Road Map TB Care ‘Aisyiyah Kabupaten Bandung, 22-23. Wai, P. P., Shewade, H. D., Kyaw, N. T. T., Thein, S., Thu, A. S., Kyaw, K. W. Y., Aye, N. N., Phyo, A. M., Maung, H. M. W., & Soe, K. T. (2018). Community-based MDR-TB care project improves treatment initiation in patients diagnosed with MDR-TB in Myanmar. PloS One, 13(3), e0194087. Kemenkes RI. (2012). Pedoman Pencegahan dan Pengendalian Tuberkulosis di Fasilitasa Kesehatan. Kesehatan, K. (2014). Profil Kesehatan Indonesia tahun 2013. PR. (2009). PR TB Care ‘Aisyiyah. Tuberkulosis dan Penanggulangannya. Modul I. Pelatihan penanggulangan Tuberkulosis bagi kader komunitas. Jakarta : PR TB ‘Aisyiyah.: Vol. I (I). Yoisangadji, M. &. (2016). Hubungan Antara Pengawas Menelan Obat (Pmo) Dan Peran Keluarga Dengan Kepatuhan Minum Obat Pada Pasien Tuberkulosis Di Wilayah Kerja Puskesmas Sario Kota Manado. Pharmacon, 5(2), 138–143. https://doi. org/10.35799/pha.5.2016.12181i Prabowo, R. D. R., & Wachidah Yuniartika, S. (2014). Hubungan Antara Peran Pengawas Minum Obat (Pmo) Dengan Kepatuhan Kunjungan Berobat Pada Pasien Tuberculosis Paru (Bb Paru) Di Puskesmas Nogosari Boyolali. Universitas Muhammadiyah Surakarta. Zulfikar. (2015). Eksistensi Program TB Care Aisyiyah Dalam pemberantasan Penyakit TB di Kab. Goa. JKA \| Volume 8 \| Nomor 2 \| Desember 2021
https://openalex.org/W3213077363	https://gfzpublic.gfz-potsdam.de/pubman/item/item_5008652_1/component/file_5008694/5008652.pdf	English	null	An interdisciplinary view of the long-term evolution of repository systems across scales: the iCROSS project	Safety of nuclear waste disposal	2,021	cc-by	1,693	safeND, 2d) Deep geological disposal – understanding processes in the multi-barrier system, BASE safeND, 2d) Deep geological disposal – understanding processes in the multi-barrier system, BASE safeND, 2d) Deep geological disp safeND, 2d) Deep geological disposal – underst Saf. Nucl. Waste Disposal, 1, 85–87, 2021 https://doi.org/10.5194/sand-1-85-2021 © Author(s) 2021. This work is distributed under the Creative Commons Attribution 4.0 License. Open Access Safety of Nuclear Waste Disposal afeND, 2d) Deep geological disposal – understanding processes in the multi-barrier system, BASE eND, 2d) Deep geological disposal – understanding processes p geological disposal – understanding processes in the multi An interdisciplinary view of the long-term evolution of repository systems across scales: the iCROSS project Dirk Bosbach1, Horst Geckeis2, Frank Heberling2, Olaf Kolditz3, Michael Kühn4, Katharina Müller5, Thorsten Stumpf5, and the iCROSS team+ 1Institut für Energie- und Klimaforschung – Nukleare Entsorgung und Reaktorsicherheit (IEK-6), Forschungszentrum Jülich (FZJ),Jülich, Germany 2Institute for Nuclear Waste Disposal (INE), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany 3Department of Environmental Informatics (ENVINF), Helmholtz Centre for Environmental Research (UFZ), Leipzig, Germany 4Deutsches GeoForschungsZentrum (GFZ), Potsdam, Germany 5Institut für Ressourcenökologie (IRE), Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden-Rossendorf, Germany +A full list appears in the team list section. Correspondence: Horst Geckeis (horst.geckeis@kit.edu) al disposal – understanding processes in the multi-barrier sys Leipzig, Germany 4Deutsches GeoForschungsZentrum (GFZ), Potsdam, Germany 5Institut für Ressourcenökologie (IRE), Helmholtz-Zentrum Dresden-Rossendorf (HZDR), Dresden-Rossendorf, Germany +A full list appears in the team list section. Correspondence: Horst Geckeis (horst.geckeis@kit.edu) g processes in the multi-barrier system, BASE Published: 10 November 2021 Abstract. The interdisciplinary project “Integrity of nuclear waste repository systems – Cross-scale system understanding and analysis (iCROSS)” combines research competencies of Helmholtz scientists related to the topics of nuclear, geosciences, biosciences and environmental simulations in collaborations overarching the re- search ﬁelds energy and earth and environment. The focus is to understand and analyze close-to-real long-term evolutionary pathways of radioactive waste repositories across nanoscales to repository scales. The project is subdivided into work packages dealing with laboratory studies, ﬁeld experiments in under- ground research laboratories (URLs), advanced modelling studies and the integration and alignment of data and information using virtual reality methods. In this sense, the project structure aims at a holistic view on relevant processes across scales in order to comprehensively simulate potential repository evolutions. Within the multi-barrier system of a repository for heat-generating radioactive waste, a number of complex reactions proceed, including dissolution, redox processes, biochemical reactions, gas evolution and solid/liquid interface and (co)precipitation reactions. At the same time, thermal and external mechanical stress has an im- pact on the conditions in a deep geological repository. All those processes are highly coupled, with multiple interdependencies on various scales and have a strong impact on radionuclide mobility and retention. In recent years, substantial progress was achieved in describing coupled thermal-hydro-mechanical-chemical-biological (THM/CB) processes in numerical simulations. A realistic and concise description of these coupled processes on different time and spatial scales is, at present, a largely unresolved scientiﬁc and computational challenge. The close interaction of experimental and simulation teams aims at a more accurate quantiﬁcation and assess- ment of processes and thus, the reduction of uncertainties and of conservative assumptions and eventually to a close-to-real perception of the repository evolution. p p p y One focus of iCROSS is directed to relevant processes in a clay rock repository. In this context, the iCROSS team became a full member of the international Mont Terri consortium and worked in close collaboration with international and German institutions in URL projects. Respective experiments speciﬁcally deal with coupled processes at the reactive interfaces in a repository near ﬁeld (e.g. the steel/bentonite and bentonite/concrete interfaces). Within iCROSS, the impact of secondary phase formation on radionuclide transport is investigated. At Mont Terri, experiments are in preparation to study radionuclide transport phenomena in clay rock formations within temperature gradients and in facies exhibiting signiﬁcant heterogeneities on different scales (nm to cm). Published by Copernicus Publications on behalf of the Federal Ofﬁce for the Safety of Nuclear Waste Management (BASE). Published: 10 November 2021 Published by Copernicus Publications on behalf of the Federal Ofﬁce for the Safety of Nuclear Waste Management (BASE). ed by Copernicus Publications on behalf of the Federal Ofﬁce for the Safety of Nuclear Waste Man 86 D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales Beside those studies, high resolution exploration methods for rock characterization are developed and tested and the effect of temperature and other boundary conditions on the strength, creep properties and healing of faults within Opalinus clay are quantiﬁed. Multiphysics models coupled to reactive transport simulation have been further developed and applied to laboratory and ﬁeld experiments. Results are digitally analyzed and illustrated in a visualization center, in order to enhance the comprehension of coupled processes in repository systems across scales. Beside those studies, high resolution exploration methods for rock characterization are developed and tested and the effect of temperature and other boundary conditions on the strength, creep properties and healing of faults within Opalinus clay are quantiﬁed. Multiphysics models coupled to reactive transport simulation have been further developed and applied to laboratory and ﬁeld experiments. Results are digitally analyzed and illustrated in a visualization center, in order to enhance the comprehension of coupled processes in repository systems across scales. The present contribution provides an overview on the project and reports selected results. The impact of considering complex coupled processes in repository subsystems for the assessment of the integrity of a given (generic) repository arrangement is discussed. Kurzfassung. Im interdisziplinären Projekt iCROSS („Integrität von Endlagern für nukleare Abfälle – Übergreifendes Systemverständnis und Analyse“) werden Forschungskompetenzen von Wissenschaftlern der Helmholtz-Institute mit den Themen Nuklearwissenschaften, Geowissenschaften, Biowissenschaften und Umweltsimulationen in einer die Forschungsbereiche Energie sowie Erde und Umwelt übergreifenden Zusam- menarbeit kombiniert. Der Fokus richtet sich darauf, realitätsnahe Langzeitentwicklungspfade von Endlagern für radioaktive Abfälle von der Nano- bis zur Endlagerskala zu verstehen und auszuwerten. Das Projekt ist unterteilt in Arbeitspakete zu Laborstudien, Feldexperimenten in unterirdischen Forschungsla- boren (URL), fortgeschrittenen Modellierungsstudien und der Integration sowie der Ausrichtung von Daten und Informationen unter Verwendung von Methoden der virtuellen Realität. In diesem Sinne zielt die Projektstruktur auf eine ganzheitliche Sicht auf relevante Prozesse über Skalen hinweg ab, um potenzielle Entwicklungen von Endlagern umfassend zu simulieren. Saf. Nucl. Waste Disposal, 1, 85–87, 2021 https://doi.org/10.5194/sand-1-85-2021 Published: 10 November 2021 The authors would like to thank all col- leagues of the iCROSS teams in the individal Helmholtz institutes for their contributions to the project. We gratefully acknowledge the help and support of all collaborating partners. Speciﬁcally we would like to thank the URL teams, notably the consortium of the Mt Terri URL, Switzerland, and our funding organisations. Financial support. This research has been supported by the Helmholtz Association (grant no. SO-093) and the Bundesminis- terium für Bildung und Forschung (grant no. 02 NUK 053 C). D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales Published: 10 November 2021 Innerhalb des Multi-Barrieren-Systems eines Endlagers für wärmeerzeugende radioaktive Abfälle laufen eine Reihe komplexer Reaktionen, einschließlich Zerfallsreaktionen, Redoxreaktionen, biochemischer Reaktionen, Gasentwicklung und Fest-Flüssig-Grenzﬂächen- sowie (Ko-)Präzipitationsreaktionen ab. Gleichzeitig haben thermische und externe mechanische Belastungen einen Einﬂuss auf die Bedingungen in einem geologischen Tiefenlager. Sämtliche dieser Prozesse sind eng miteinander verbunden – mit multiplen Interdependenzen auf verschiedenen Ebenen – und haben einen großen Einﬂuss auf die Radionuklidmobilität und -retention. In den ver- gangenen Jahren wurden wesentliche Fortschritte bei der Beschreibung gekoppelter thermo-hydromechanisch- chemisch-biologischer (THM/CB-)Prozesse in numerischen Simulationen erzielt. Eine realistische Beschrei- bung dieser gekoppelten Prozesse auf unterschiedlichen Zeit- und Raumskalen ist zum gegenwärtigen Zeit- punkt eine großenteils ungelöste wissenschaftliche und rechnerische Herausforderung. Die enge Interaktion von Experimentier- und Simulationsteams zielt auf eine genauere Quantiﬁzierung und Beurteilung von Prozessen ab und somit auf die Verminderung von Ungewissheiten und von konservativen Annahmen und schließlich auf eine realitätsnahe Wahrnehmung der Entwicklung des Endlagers. g g g Ein Fokus von iCROSS richtet sich auf relevante Prozesse in einem Tongestein-Endlager. In diesem Zusammenhang wurde das iCROSS-Team Vollmitglied des internationalen Mont-Terri-Konsortiums und ar- beitet in enger Kooperation mit internationalen und deutschen Institutionen in URL-Projekten zusammen. Die entsprechenden Experimente befassen sich insbesondere mit gekoppelten Prozessen an den reaktiven Grenzﬂächen im Nahfeld eines Endlagers (z. B. die Stahl-Bentonit- und Bentonit-Beton-Grenzﬂächen). Inner- halb von iCROSS werden die Auswirkungen der Sekundärphasenbildung auf den Radionuklidtransport unter- sucht. Im Mont-Terri-Labor beﬁnden sich Experimente in Vorbereitung, um Radionuklidtransportphänomene in Tongesteinsformationen innerhalb von Temperaturgradienten und angesichts erheblicher Heterogenitäten in un- terschiedlichen Maßstäben (nm bis cm) zu untersuchen. Neben diesen Untersuchungen werden hochauﬂösende Untersuchungsmethoden für die Gesteinscharakterisierung entwickelt und geprüft, und die Auswirkungen von Temperatur und anderen Randbedingungen auf die Festigkeit, Kriecheigenschaften und Heilung von Verwerfun- gen innerhalb von Opalinus-Ton werden quantiﬁziert. Multiphysikalische Modelle in Kombination mit reaktiver Transportsimulation werden weiterentwickelt worden und in Labor- sowie Feldexperimenten eingesetzt. Die Ergebnisse werden digital ausgewertet und in einem Visualisierungszentrum dargestellt, um das Verständnis gekoppelter Prozesse in Endlagersystemen skalenübergreifend zu verbessern. Der vorliegende Beitrag bietet einen Überblick über das Projekt und legt ausgewählte Ergebnisse dar. Die Auswirkungen der Betrachtung komplexer gekoppelter Prozesse in Endlager-Subsystemen auf die Beurteilung der Integrität einer gegebenen exemplarischen Endlageranordnung werden erörtert. Saf. Nucl. Waste Disposal, 1, 85–87, 2021 https://doi.org/10.5194/sand-1-85-2021 D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales 87 Team list. The iCROSS team consists of individal Helmholtz in- stitutes. Acknowledgements. D. Bosbach et al.: An interdisciplinary view of the long-term evolution of repository systems across scales 87 Financial support. This research has been supported by the Helmholtz Association (grant no. SO-093) and the Bundesminis- terium für Bildung und Forschung (grant no. 02 NUK 053 C). Team list. The iCROSS team consists of individal Helmholtz in- stitutes. Acknowledgements. The authors would like to thank all col- leagues of the iCROSS teams in the individal Helmholtz institutes for their contributions to the project. We gratefully acknowledge the help and support of all collaborating partners. Speciﬁcally we would like to thank the URL teams, notably the consortium of the Mt Terri URL, Switzerland, and our funding organisations. https://doi.org/10.5194/sand-1-85-2021 Saf. Nucl. Waste Disposal, 1, 85–87, 2021
https://openalex.org/W3046709342	https://www.researchsquare.com/article/rs-44741/latest.pdf	English	null	Characterization of volatile fatty-acid utilization in Escherichia coli aiming for robust valorisation of food residues	AMB express	2,020	cc-by	7,123	Characterization of volatile fatty-acid utilization in Escherichia coli aiming for robust valorisation of food residues Gustav Sjöberg KTH Royal Institute of Technology Martin Gustavsson KTH Royal Institure of Technology Antonius J.A. van Maris (  tonvm@kth.se ) KTH Royal Institute of Technology https://or Original article Version of Record: A version of this preprint was published at AMB Express on October 17th, 2020. See the published version at https://doi.org/10.1186/s13568-020-01121-4. Page 1/17 Page 1/17 Abstract Valorisation of food residues would greatly benefit from development of robust processes that create added value compared to current feed- and biogas applications. Recent advances in membrane- bioreactor-based open mixed microbial cultures, enable robust conversion of fluctuating streams of food residues to a mixture of volatile fatty acids (VFAs). In this study, such a mixed stream of VFAs was investigated as a substrate for Escherichia coli, a well-studied organism suitable for application in further conversion of the acids into compounds of higher value, and/or that are easier to separate from the aqueous medium. E. coli was cultured in batch on a VFA-rich anaerobic digest of food residues, tolerating up to 40 mM of total VFAs without any reduction in growth rate. In carbon-limited chemostats of E. coli W3110 ΔFadR on a simulated VFA mixture, the straight-chain VFAs (C2-C6) in the mixture were readily consumed simultaneously. At a dilution rate of 0.1 h− 1, mainly acetic-, propionic- and caproic acid were consumed, while consumption of all the provided acids were observed at 0.05 h− 1. Interestingly, also the branched isovaleric acid was consumed through a hitherto unknown mechanism. In total, up to 80% of the carbon from the supplied VFAs was consumed by the cells, and approximately 2.7% was excreted as nucleotide precursors in the medium. These results suggest that VFAs derived from food residues are a promising substrate for E. coli. Volatile fatty acids from food waste are a suitable carbon source for E. coli Consumption of all supplied volatile fatty acids was observed in chemostats Volatile fatty acids from food waste are a suitable carbon source for E. coli Consumption of all supplied volatile fatty acids was observed in chemostats Volatile fatty acids from food waste are a suitable carbon source for E. c Volatile fatty acids from food waste are a suitable carbon source for E. coli Introduction vinyl- and cellulose esters), pharmaceuticals, fragrances and flavours (i.e. various esters), food and feed supplements, crop protection and disinfecting products (Kubitschke et al. 2014). Recent advances in application of membrane bioreactors for anaerobic digestion have simplified the recovery of VFAs from the broth, by allowing continuous removal of a clear liquid fraction from the solids (Trad et al. 2015; Wainaina et al. 2019). However, the total VFA concentration in the effluent remains low, and thus distillation or extraction of the individual acids cannot compete with current, fossil-based production methods (Kleerebezem et al. 2015). As an alternative to purifying VFAs for direct use, they can be further valorised to a set of valuable products by microbial conversion (Kleerebezem et al. 2015). Compounds that can be readily produced include, but are not limited to: polymers, alcohols, hydroxyacids, lipids and esters (Chang et al. 2010; Lee et al. 2014). Such conversions allow VFAs to become a platform intermediate that eliminates the variation of food residues, while preserving most of the contained electrons of this complex substrate. Unlike the original food residues, the resulting VFA mix is composed of a fixed set of compounds with potential as a carbon- and energy source for pure cultures of microorganism able to produce value-added chemicals, as well as direct microbial conversion into value added compounds. ( ) concentration in the effluent remains low, and thus distillation or extraction of the individual acids cannot compete with current, fossil-based production methods (Kleerebezem et al. 2015). As an alternative to purifying VFAs for direct use, they can be further valorised to a set of valuable products by microbial conversion (Kleerebezem et al. 2015). Compounds that can be readily produced include, but are not limited to: polymers, alcohols, hydroxyacids, lipids and esters (Chang et al. 2010; Lee et al. 2014). Such conversions allow VFAs to become a platform intermediate that eliminates the variation of food residues, while preserving most of the contained electrons of this complex substrate. Unlike the original food residues, the resulting VFA mix is composed of a fixed set of compounds with potential as a carbon- and energy source for pure cultures of microorganism able to produce value-added chemicals, as well as direct microbial conversion into value added compounds. Escherichia coli is a well-established bacterial host and has been studied extensively for proof-of-concept production of numerous compounds (Calero and Nikel 2019). Introduction Approximately one third of the food produced for humans is lost as waste before consumption (FAO 2011). Together with actions to decrease these losses, valorisation of the inevitable remaining residues is preferred over dumping in landfills, where anaerobic bacteria decompose the waste into the potent greenhouse-gas methane. Similar natural consortia of bacteria can also convert a wide-range of different, fluctuating food residues to methane under more controlled conditions in a process called anaerobic digestion (Keller and Surette 2006; Brenner et al. 2008; Smid and Lacroix 2013). The resulting biogas is a drop-in replacement for natural gas, providing energy for propulsion, electricity or heat (Scarlat et al. 2018). However, the value of biogas is relatively low and is often subsidised to benefit from the reduction in waste volumes and greenhouse gas emissions in addition to the production of green energy and fuels (Bartolini et al. 2017). Like the open mixed cultures of anaerobic digestion, alternative processes for valorising food residues must be robust and able to cope with the fluctuations of the incoming residues, while yielding stable value-added product streams. This eliminates the use of pure cultures, since they cannot deal with the complex and fluctuating compositions of mixed food waste and are additionally economically undesirable and difficult to sterilize. Although production of methane is thermodynamically favoured, by Page 2/17 operating anaerobic digestion under conditions that are unfavourable to methanogens, such as reduced pH, reduced hydraulic retention time, or increased temperature (Lee et al. 2014), formation of volatile fatty acids (VFAs) is promoted instead. Volatile fatty acids (VFAs) are carboxylic acids with between two and six carbon atoms, produced as intermediates during digestion of organic material and have higher-value applications than the otherwise produced biogas. These applications include polymers (e.g. vinyl- and cellulose esters), pharmaceuticals, fragrances and flavours (i.e. various esters), food and feed supplements, crop protection and disinfecting products (Kubitschke et al. 2014). Recent advances in application of membrane bioreactors for anaerobic digestion have simplified the recovery of VFAs from the broth, by allowing continuous removal of a clear liquid fraction from the solids (Trad et al. 2015; Wainaina et al. 2019). However, the total VFA Volatile fatty acids (VFAs) are carboxylic acids with between two and six carbon atoms, produced as intermediates during digestion of organic material and have higher-value applications than the otherwise produced biogas. These applications include polymers (e.g. Introduction In addition to the wide variety of possible (engineered) products, E. coli consumes a wide range of substrates, including all the straight chain fatty acids from two to six carbons (Salanitro and Wegener 1971). However, anaerobic digestion produces a mixture of these VFAs and the simultaneous consumption of these has not been previously characterized in E. coli. Additionally, the limits of tolerance of E. coli to this mixture of weak organic acids, which can diffuse through the membrane and uncouple the membrane potential (Baronofsky et al. 1984; Gabba et al. 2020), is unknown. The aim of this study was to investigate the potential of Escherichia coli as a platform microorganism for valorisation of the relatively clean and defined VFA streams originating from membrane-bioreactor-based anaerobic digestion (Wainaina et al. 2019). Determination of the requirements for co-consumption of VFAs by E. coli is a first step towards engineering the subsequent metabolism for their valorisation. First, the limits of tolerance, (co)consumption and growth of E. coli on a VFA solution from an anaerobic digest of food residues was characterized in batch shake flask cultures. Subsequently, co-consumption of the VFAs was investigated in carbon-limited chemostat cultures. Strains To assess VFA consumption and tolerance of E. coli, the reference strain W3110 (ATCC® 27325™, F− lambda− IN(rrnD-rrnE)1 rph-1) and the W3110-derived W3110 ΔFadR were used. W3110 ΔFadR contains an in-frame deletion of the transcriptional regulator FadR, and was kindly provided by Prof. Sang Yup Lee. Both strains were stored at -80 °C in minimal salts medium with 5 g L− 1 glucose and 25% glycerol. Methods Page 3/17 Page 3/17 Page 3/17 Minimal salt media All experiments were carried out in minimal salts medium consisting of 5 g L− 1 (NH4)2SO4 (Merck, Darmstadt, Germany), 1.6 g L− 1 KH2PO4 (VWR International, Leuven, Belgium), 0.5 g L− 1 diammonium citrate (Merck) and 6.6 g L− 1 Na2HPO4·2H2O (VWR International), which was sterilized by autoclaving for 20 minutes at 121 °C. 1M MgSO4 and trace element stock solution (Sandén et al. 2003) were autoclaved separately (20 minutes at 121 °C) and added to a final amount of 1 mL per liter minimal salts medium each. Carbon sources were added as described in the following sections. Chemostat cultivations on defined medium with VFAs as carbon source A defined medium with the same distribution of VFAs as measured in the anaerobic digest was designed for chemostat experiments, thereby circumventing volumetric limitations of the available digest from Wainaina et al. (2019). Each carboxylic acid was added directly to 80 L of previously autoclaved minimal salts medium, to final concentrations of 860 mg L− 1 acetic acid, 170 mg L− 1 propionic acid, 640 mg L− 1 butyric acid, 580 mg L− 1 isovaleric acid, 100 mg L− 1 valeric acid and 2500 mg L− 1 caproic acid (≥ 99%, Sigma-Aldrich, St. Louis, USA). The minimal medium was set to pH 7.0 by addition of 1.59 g L− 1 of NaOH (Merck). Chemostats were run in a parallel system with 6 steam-sterilized stainless-steel stirred-tank bioreactors (GRETA, Belach Bioteknik, Skogås, Sweden). The connections to the medium tank were made with autoclaved silicon tubing through the integrated peristaltic pump heads. Each inlet pump was calibrated daily to the desired flow-rate using an inline burette, that could be filled by aspiration through a sterile air filter (Filtropur S, Sarstedt, Nümbrecht, Germany). The volume of each bioreactor was automatically maintained at 800 mL by activation of the outlet pumps by conductivity-based level sensors. Each outlet tube was attached to a sterile 20L plastic bottle for aseptic collection of broth (Nalgene, Thermo Fisher Scientific). The pH was maintained at 7.0 by automated titration with 4 M H2SO4. Starting at 500 rpm and 200 mL min− 1 in the batch phase, the stirring speed and air flow were increased as required to maintain a dissolved oxygen tension above 30% during the chemostat phase. The cultures were monitored by aseptically withdrawing 20 ml samples with a syringe for determination of cell dry weight (CDW), OD600 and supernatant composition. The outlet gas from each reactor was connected through a multiplexer (Belach Bioteknik) to a 1313 Fermentation Monitor (LumaSense Technologies, Santa Clara, USA) for determination of CO2 concentrations. Inoculum for all 6 bioreactors was prepared by transfer of a W3110 ΔFadR freezer stock to a sterile stainless-steel stirred-tank bioreactor (Belach Bioteknik) containing 5.0 L of minimal medium supplemented with 5 g L− 1 of glucose. The stirrer speed was set to 1000 rpm and 5.0 L min− 1 headspace airflow was applied. Shake flask cultivations with an anaerobic digest as carbon source Anaerobic digest was provided by Prof. Taherzadeh (Borås University, Sweden) in two plastic bottles containing 3 L of clarified food residue digest from a previous experiment (Wainaina et al. 2019). Upon arrival, the solution was thawed, and the liquid was decanted from a rust-brown flaky precipitate, which had also been observed by Wainaina et al. (2019) after storage of the solution at -20 °C (personal communication). The clarified solution was aliquoted in smaller glass bottles and refrozen until use. Upon rethawing, the aliquots were adjusted to pH 7.0 with 1M NaOH and then filter-sterilized through bottle-top 0.22 µm polyethersulfone filters (Corning, New York, USA). The sterile digest was diluted in autoclaved deionized water and a 10X concentrate of the minimal medium salts. For the control experiment without minimal medium, 10% anaerobic digest and 90% 88 mM MOPS (AppliChem, Darmstadt, Germany) were mixed (v/v). Cells were prepared by thawing glycerol stocks of the desired strains and inoculating to minimal salts medium, supplemented with 5 g L− 1 glucose (Thermo Fisher Scientific, Waltham, USA) from an autoclaved stock solution. After overnight incubation at 37 °C with 180 rpm shaking (Minitron HT Infors, Bottmingen-Basel, Switzerland), the cells had reached an optical density at 600 nm (OD600) between 1.0 and 2.0. Exponentially growing cells were harvested by centrifugation at 3000 g for 10 minutes (Avanti J- 20 XP, Beckman Coulter, Brea, USA) and resuspended in the various dilutions of anaerobic digest. Baffled shake flasks were filled to 10% of their maximum volume to ensure sufficient oxygen transfer, and Page 4/17 incubated at 37 °C with 180 rpm shaking. Samples were withdrawn regularly for determination of OD600 and analysis of the medium by high-performance liquid chromatography (HPLC). Chemostat cultivations on defined medium with VFAs as carbon source After overnight culture, the broth of the exponentially-growing culture (with an OD600 between 1.0–2.0) was centrifuged aseptically at 3000 g for 10 minutes (Sorvall BIOS 16, Thermo Fisher Scientific), then resuspended in the chemostat medium with VFA before inoculation by syringe to each of the 6 bioreactors. Analyses Page 5/17 The concentrations of phosphate and ammonia in the anaerobic digest were determined spectrophotometrically using a Cedex Bio Analyzer (Roche Diagnostics, Mannheim, Germany) according to the manufacturer’s instructions. OD600 was measured in a spectrophotometer (Genesys 20, Thermo Fisher Scientific) at 600 nm after dilutions to OD600 between 0.1 and 0.2 with a 9 g L− 1 NaCl solution. CDW was measured in triplicate for each sample point. 5 mL of medium was added to dried and pre- weighed glass tubes, weighed and then centrifuged at 2500 g in a tabletop centrifuge (Z206 A, Hermle, Gosheim, Germany) for 10 minutes, washed once with 5 mL of 9 g L− 1 NaCl, centrifuged again and dried overnight. The CDW (g L− 1) was determined by dividing the dry weight of the cells by the volume of the original broth sample. HPLC was used to analyze the composition of culture supernatants, using an Alliance 2695 system (Waters, Milford, MA, USA) equipped with a 2414 refractive index detector (Waters), a 2996 photodiode array detector (Waters), and a TCM column heater (Waters). VFAs were separated on an Aminex HPX-87H organic acid column (Bio-Rad, Hercules, CA, USA) with a mobile phase containing 0.2% phosphoric acid and 5% acetonitrile in MilliQ water, at a flow rate of 0.9 mL min− 1. The column was maintained at 85 °C and the acids were quantified with the photodiode detector at set at 210 nm (Bell et al. 1991). Orotic acid, uracil, and thymine were separated on the same column but at room temperature, with a flow of 0.5 ml min− 1, and with 0.008 N sulfuric acid as mobile phase. The UV spectrum, refractive index and retention times of the reference compounds were used for identification, and peaks at 210 nm were used for quantification. Tricarboxylic-acid-cycle intermediates and common E. coli fermentation products were quantified based on previously determined retention times and absorbance spectra. Preparative HPLC was performed under the same conditions, but with the maximum possible injection volume, 200 µL. Fractions were collected manually as they exited the photodiode detector. Each fraction of interest was dried under vacuum and dissolved in 1 mL D2O (Merck), before analysis by nuclear magnetic resonance (NMR). Spectra were obtained at 400 MHz with 128 scans on a Bruker Avance (Bruker Corporation, Billerica, USA). Amino acids were assayed using the AccQ-Tag method (Waters) with proprietary reagents, according to the manufacturer’s instructions. Analyses Nucleotides and nucleosides were analyzed according to the protocol developed by Childs et al. (Childs et al. 1996). Results To investigate the requirement for additional nutrients for cultivation of E. coli, the concentrations of VFAs, free phosphate, and free ammonia in the sample of clarified anaerobic digest of food residues were measured (Table 1). This identified that the low amounts of free phosphate and ammonia would be growth limiting compared to the VFA content. Thus, the digest was supplemented with minimal salts medium to ensure that the VFA content would be the limiting factor. Page 6/17 Page 6/17 Table 1 Measured concentrations of key components of the anaerobic digest used in shake flask batch cultures of E. coli. Component Concentration (g L− 1) Acetic acid 0.81 Propionic acid 0.13 Butyric acid 0.53 Isovaleric acid 0.49 Valeric acid 0.09 Caproic acid 2.01 Free phosphate 0.018 Free ammonia 0.123 Table 1 Tolerance to and consumption of VFAs from anaerobic digest by E. coli To assess its tolerance to the anaerobic digest, the VFA constituents of which are known growth inhibitors, E. coli W3110 was grown in shake flasks with dilutions of the digest (Fig. 1a). The strain grew on the anaerobic digest across the range of concentrations tested (Fig. 1a), indicating that E. coli is sufficiently robust towards the VFA concentrations in the current generation of membrane bioreactor anaerobic digest (Wainaina et al. 2019). As expected from the composition of the digest, W3110 did not show significant growth in a control experiment lacking minimal salts medium supplementation (10%+MOPS in Fig. 1a). After approximately 10 hours, growth started to decrease and significant amounts of VFAs remained unconsumed by W3110 (Fig. 1b and 1c). To de-repress pathways involved in fatty acid degradation and possibly improve VFA consumption, the fatty acid degradation transcriptional regulator (encoded by fadR) was deleted, and the resultant strain W3110 ΔFadR was compared to the W3110 wild type (Fig. 1). Over the first 10 hours, W3110 ΔFadR showed similar tolerance to the anaerobic digest (Fig. 1a) and achieved similar initial growth rates (Fig. S2). Interestingly, W3110 ΔFadR maintained growth for a longer time and reached a higher final OD600. This can in part be explained by a higher uptake of valeric acid and caproic acid (Fig. 1b and 1c), due to the lack of repression of fatty acid catabolism. However, the final OD600 of W3110 ΔFadR increased more than can solely be explained by the increased consumption of valeric- and caproic acid (Fig. 1a), which might be caused by consumption of a hitherto unidentified compound during the last 20 hours of cultivation. Although W3110 ΔFadR consumed a larger fraction of the VFAs than W3110, consumption of valeric- and caproic acid was not complete and W3110 did not consume butyric- and isovaleric acid. Page 7/17 Page 7/17 Chemostat culture of E. coli on VFAs Carbon-limited chemostat cultures are known to promote co-consumption of various carbon sources that in batch would either be consumed sequentially or not at all (Egli et al. 1993). To improve consumption of the VFAs present in the anaerobic digest (Table 1) and to characterize the co-consumption of VFAs in E. coli more thoroughly, the behaviour of W3110 ΔFadR was investigated in carbon (VFA)-limited chemostat cultures at dilution rates of (approximately) 0.05 and 0.10 h− 1. Since the amounts available of the original material were insufficient, the chemostat culture was performed on a defined medium with a VFA distribution identical to the anaerobic digest (Table 1). During the start-up batch phase of the culture, it was observed that the growth rate in the defined medium was 0.140 ± 0.005 h− 1, which is lower the initial growth rate of 0.2 h− 1 observed in the anaerobic digest (Fig. S1). The lower of the two dilution rates took more than the theoretical 5 volume changes to reach steady state, and the volume varied significantly due to foaming, which triggered the level sensor-activated outlet pump and caused a rather large standard deviation (15% relative standard deviation) on the dilution rate between replicates (Table 2). Page 8/17 Table 2 Summary of values from chemostat cultivations of W3110 ΔFadR on a defined minimal salts medium with a mix of VFAs as carbon source. The medium composition was designed from measurements of an anaerobic digest of food residues. Chemostat culture of E. coli on VFAs D (h− 1) 0.054 ± 0.008 0.094 ± 0.001 CDW (g L− 1) 2.06 ± 0.05 0.66 ± 0.05 YX/S (g g− 1) 0.53 ± 0.02 0.48 ± 0.04 qCO2 (mmol g− 1 h− 1) 2.2 ± 0.4 3.9 ± 0.3 qOrotic acid (mmol g− 1 h− 1) 0.026 ± 0.003 0.039 ± 0.003 Carbon balance 92% ± 7% 103% ± 6% mmol g− 1 h− 1 mmol DRa g− 1 h− 1 mmol g− 1 h− 1 mmol DRa g− 1 h− 1 qAcetic acid 0.38 ± 0.05 3.0 ± 0.4 2.1 ± 0.2 16 ± 1 qPropionic acid 0.051 ± 0.003 0.72 ± 0.04 0.24 ± 0.03 3.4 ± 0.4 qButyric acid 0.08 ± 0.02 1.5 ± 0.3 NDb NDb qIsovaleric acid 0.009 ± 0.002 0.25 ± 0.06 NDb NDb qValeric acid 0.025 ± 0.003 0.66 ± 0.08 NDb NDb qCaproic acid 0.57 ± 0.08 18 ± 2 0.46 ± 0.05 15 ± 1 qTotal VFA 4.8 ± 0.6 c 24 ± 3 7.6 ± 0.6 c 35 ± 4 Averages ± standard deviations. n = 6 (biological triplicates each sampled at two different timepoints separated by at least 1/D hours) Table 2 At a growth rate of 0.094 h− 1, about 90% of the consumed electrons and carbon came from acetic- and caproic acid, while the remainder originated from propionic acid. At this dilution rate, only 23% ± 1% of Page 9/17 Page 9/17 Page 9/17 the VFAs were consumed on a carbon basis. Whilst all the acetate was consumed, only 14% ± 1% of caproate and 73% ± 6% of propionate was consumed, whilst butyric-, isovaleric- and valeric acid were not consumed at all (Fig. 2). At 0.094 h-1, the consumption rate of VFAs was 7.6 ± 0.6 mmolcarbon g− 1 h− 1, which resulted in 0.66 ± 0.05 g CDW at a biomass yield on VFA of 0.48 ± 0.04 g g− 1. the VFAs were consumed on a carbon basis. Whilst all the acetate was consumed, only 14% ± 1% of caproate and 73% ± 6% of propionate was consumed, whilst butyric-, isovaleric- and valeric acid were not consumed at all (Fig. 2). At 0.094 h-1, the consumption rate of VFAs was 7.6 ± 0.6 mmolcarbon g− 1 h− 1, which resulted in 0.66 ± 0.05 g CDW at a biomass yield on VFA of 0.48 ± 0.04 g g− 1. Chemostat culture of E. coli on VFAs The biomass yield on consumed substrate (YX/S) at a dilution rate of 0.054 h− 1 was 0.53 ± 0.02 g g− 1, which was not significantly different from the higher dilution rate. However, the CDW was about threefold higher at 2.06 ± 0.05 g L− 1 for the dilution rate of 0.054 h− 1 (Table 2). This difference is mainly explained by a higher total carbon consumption due to improved co-utilization of butyric- (44% ± 9% consumed), valeric- (97% ± 0% consumed), and caproic acid (99% ± 1% consumed) at the lower dilution rate (Fig. 2). At this lower dilution rate, the specific consumption rates of acetic- and propionic acid were reduced, due to both the lower dilution rate as well as the co-consumption of additional carbon sources (Table 2). In these conditions, most of the electrons were derived from caproic acid, while the shorter chain acids together contributed less than 25% of the degree of reduction (DR) (Heijnen 1994). Despite the increased consumption of butyric acid at 0.054 h− 1, a substantial amount remained in the medium. In addition, at a growth rate of 0.054 h− 1 W3110 ΔFadR consumed isovaleric acid at a significant, albeit slow, rate (Table 2). However, the contribution of this compound to the total carbon consumed was minimal (< 1% of total carbon). Although the carbon balance only closed to 90% ± 7% at the lower dilution rate, supernatant samples tested negative for the presence of amino acids, commonly occurring fermentation products and citric acid cycle intermediates. Instead, a set of unknown peaks was detected by refractive index and UV absorbance on the carbohydrate- and organic acid column. The chromatography was repeated on a preparative scale for one of the samples and interesting fractions were collected. Based on the NMR spectra of one of the unknown fractions, it was possible to identify orotic acid as a by-product, contributing to approximately 2.7% of the carbon at both dilution rates, which improved the carbon balance to 92% ± 7%. Since orotic acid is an intermediate of nucleotide synthesis, other compounds belonging to these pathways were also sought. Thereby, trace levels of thymine and uracil were identified in the medium samples, though they did not contribute significantly to the carbon balance. Discussion The aim of this study was to evaluate the use of a clarified anaerobic digest rich in VFAs as a carbon- and energy source for E. coli. As a first step, it was established that the toxic effects of up to 40 mM mixed VFAs, as found in the anaerobic digest, were not significant at pH 7, which is in line with previous observations on the impact of rumen VFAs (Wolin 1969) or C6-C10 carboxylic acids on E. coli (Royce et al. 2013). Interestingly, the observation of a higher growth rate on the anaerobic digest (0.2 h− 1) compared to the growth on defined VFA medium in the batch phase preceding the chemostat culture (0.14 h− 1), is also in line a previously observed growth stimulation by rumen fluid (Wolin 1969). These observations Page 10/17 Page 10/17 suggest that both rumen fluid and the anaerobic digest contain unidentified growth-promoting compounds. suggest that both rumen fluid and the anaerobic digest contain unidentified growth-promoting compounds. Consumption of as many of the available VFAs as possible is another important aspect of valorisation of the VFAs from anaerobic digests. In the absence of long chain acyl-CoA (C14-C18), which is expected in the anaerobic digest, FadR represses the genes encoding the fatty acid degradation pathway (Overath et al. 1969; Cronan and Subrahmanyam 1998). By deleting FadR, the fatty acid degradation pathway, which is known to be able to beta-oxidize acids with a chain length as low as four carbon atoms (Binstock et al. 1977), was de-repressed and thereby significantly increased the fraction of VFAs that was consumed. Co- consumption of VFAs was further improved in carbon-limited chemostats, where a dilution rate of 0.054 h− 1 allowed at least partial consumption of all the acids. Given the long time needed to achieve steady state, it is likely that adaption contributed to the additional consumption of caproic acid, and to a lesser extent also isovaleric- and butyric acid, at the lower dilution rate, whilst this was not observed at 0.094 h− 1. The significantly higher caproic acid consumption rate at 0.054 h− 1 compared to 0.094 h− 1 (p = 0.01, n = 6), indicates that capacity of the uptake system was not the main limiting factor. Discussion This makes it likely that activation of the free fatty acid to the acyl-CoA was limiting the consumption of these VFAs at the higher dilution rates, as previously shown for low expression of acetyl-CoA- transferases with specificity towards butyric acid (Pauli and Overath 1972). The overall increased fraction of VFAs resulted in a higher CDW, which correlates well with previous chemostat cultures on mixed substrates (Egli et al. 1993). At the lowest dilution rate, 75% of VFAs expressed as degree of reduction came from the consumed caproic acid, whilst this was equally shared between acetic acid and caproic acid at the higher dilution rate. The Yx/s expressed in g biomass formed per g VFA consumed was higher at the lower dilution rate, which seems to contradict the impact of the maintenance requirement on the biomass yield (Pirt and Hinshelwood 1965). However, when corrected for the higher degree of reduction (and thereby energy density) of caproic acid, the Yx/s was 2.22 ± 0.07 g CDW molDR −1 at 0.054 h− 1 versus 2.7 ± 0.2 g CDW molDR −1 at 0.094 h− 1. This reflects the increased impact of maintenance at the lower dilution rate. The VFA limited chemostat also yielded two unexpected results not previously observed for E. coli: Consumption of isovaleric acid and secretion of orotic acid. The low, but statistically significant (p = 0.0002), isovaleric acid consumption exceeds quantities that at a pH of 7 can simply accumulate intracellularly or be adsorbed by cell constituents. One or more of the acetyl-CoA-transferases expressed in the W3110 ΔFadR strain under VFA-limited conditions might show promiscuity towards isovaleric acid to produce isovaleryl-CoA. However, no dedicated isovaleryl-CoA degradation pathways are known in E. coli, which would require further metabolism or conversion to a currently unidentified compound through other promiscuous enzyme activities, such as the previously shown reduction of isovaleryl-CoA to 3- methylcrotonyl-CoA by AidB (Rohankhedkar et al. 2006). Although excretion of a wide range of nucleobases by E. coli, such as thymidine, thymine, uracil, cytosine, and guanine, has previously been observed after selection in chemostat cultures (Tsen 1994), excretion of orotic acid was not previously observed. Since orotic acid itself, and the pyrimidines derived from it, can be valuable products, it may be Page 11/17 Page 11/17 interesting to investigate the metabolic pathway leading to its formation and further engineer E. coli to improve the flux towards the product. Discussion The ability to tolerate and grow in the anaerobic digest and to co-consume the VFAs, makes E. coli an interesting host for transforming VFAs to more valuable compounds and thereby valorise a diverse range of residue streams. Deletion of FadR increased the range of VFAs that were consumed in batch conditions and application of carbon-limited conditions with VFAs as the sole carbon- and energy source in chemostats showed that all substrates could (to some extent) be co-consumed. This makes it likely that the slow growth and low consumption of butyrate and isovalerate can be alleviated by laboratory evolution, for example in repeated batch- or chemostat cultivation on the substrate mix, or by engineering of the VFA consumption pathways. Page 12/17 Abbreviations • CDW • Cell dry weight • OD600 • Optical density at 600 nm • VFAs • Volatile fatty acids • HPLC • High performance liquid chromatography • NMR • Nuclear magnetic resonance • FadR • Fatty acid degradation transcriptional regulator (encoded by fadR) • YX/S • The yield of cells on substrate • DR • Degree of reduction Declarations Ethics approval and consent to participate Not applicable Consent for publication Not applicable Page 12/17 Abbreviations • CDW • Cell dry weight • OD600 • Optical density at 600 nm • VFAs • Volatile fatty acids • HPLC • High performance liquid chromatography • NMR • Nuclear magnetic resonance • FadR • Fatty acid degradation transcriptional regulator (encoded by fadR) • YX/S • The yield of cells on substrate • DR • Degree of reduction Declarations Ethics approval and consent to participate Not applicable Consent for publication Not applicable Author contributions The authors conceived the study and designed the experiments together. GS performed the experiments, processed the data and drafted the manuscript. All authors analyzed the data, revised the first draft and approved the final manuscript. Funding This work was funded by the Swedish research council Formas (grant numbers 211-2013-70 and 2014 − 1620). Acknowledgements We would like to thank Emil Sundäng Peters and Viktor Westerlund for their exploratory work that led up to the conception of this study, Maja Finnveden for her assistance with the NMR analysis, Prof. Sang Yup Lee for providing the E. coli strain W3110 ΔFadR, and Steven Wainaina and Prof. Mohammad Taherzadeh for providing the anaerobic digest used in the study. GS declares that he has no conflict of interest. AvM declares that he has no conflict of interest. Consent for publication Not applicable Page 12/17 Page 12/17 References Egli T, Lendenmann U, Snozzi M (1993) Kinetics of microbial growth with mixtures of carbon sources. 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Chang HN, Kim N-J, Kang J, Jeong CM (2010) Biomass-derived volatile fatty acid platform for fuels and chemicals. Biotechnol Bioprocess Eng 15:1–10. https://doi.org/10.1007/s12257-009-3070-8 7. Chang HN, Kim N-J, Kang J, Jeong CM (2010) Biomass-derived volatile fatty acid platform for fuels and chemicals. Biotechnol Bioprocess Eng 15:1–10. https://doi.org/10.1007/s12257-009-3070-8 8. Childs KF, Ning X-H, Bolling SF (1996) Simultaneous detection of nucleotides, nucleosides and oxidative metabolites in myocardial biopsies. J Chromatogr B Biomed Sci App 678:181–186. https://doi.org/10.1016/0378-4347(95)00474-2 8. Childs KF, Ning X-H, Bolling SF (1996) Simultaneous detection of nucleotides, nucleosides and oxidative metabolites in myocardial biopsies. J Chromatogr B Biomed Sci App 678:181–186. https://doi.org/10.1016/0378-4347(95)00474-2 9. Cronan JE, Subrahmanyam S (1998) FadR, transcriptional co-ordination of metabolic expediency. Mol Microbiol 29:937–943 9. Cronan JE, Subrahmanyam S (1998) FadR, transcriptional co-ordination of metabolic expediency. Mol Microbiol 29:937–943 10. Egli T, Lendenmann U, Snozzi M (1993) Kinetics of microbial growth with mixtures of carbon sources. Antonie Van Leeuwenhoek 63:289–298. https://doi.org/10.1007/BF00871224 10. References Pauli G, Overath P (1972) ato operon: a highly inducible system for acetoacetate and butyrate degradation in Escherichia coli. Eur J Biochem 29:553–562. https://doi.org/10.1111/j.1432- 1033.1972.tb02021.x 19. Pauli G, Overath P (1972) ato operon: a highly inducible system for acetoacetate and butyrate degradation in Escherichia coli. Eur J Biochem 29:553–562. https://doi.org/10.1111/j.1432- 1033.1972.tb02021.x 20. Pirt SJ, Hinshelwood CN (1965) The maintenance energy of bacteria in growing cultures. Proc R Soc Lond B Biol Sci 163:224–231. https://doi.org/10.1098/rspb.1965.0069 20. Pirt SJ, Hinshelwood CN (1965) The maintenance energy of bacteria in growing cultures. Proc R Soc Lond B Biol Sci 163:224–231. https://doi.org/10.1098/rspb.1965.0069 Page 14/17 21. Rohankhedkar MS, Mulrooney SB, Wedemeyer WJ, Hausinger RP (2006) The AidB component of the Escherichia coli adaptive response to alkylating agents is a flavin-containing, DNA-binding protein. J Bacteriol 188:223–230. https://doi.org/10.1128/JB.188.1.223-230.2006 22. Royce LA, Liu P, Stebbins MJ, Hanson BC, Jarboe LR (2013) The damaging effects of short chain fatty acids on Escherichia coli membranes. Appl Microbiol Biotechnol 97:8317–8327. https://doi.org/10.1007/s00253-013-5113-5 23. Salanitro JP, Wegener WS (1971) Growth of Escherichia coli on short-chain fatty acids: growth characteristics of mutants. J Bacteriol 108:885–892 24. Sandén AM, Prytz I, Tubulekas I, Förberg C, Le H, Hektor A, Neubauer P, Pragai Z, Harwood C, Ward A, Picon A, De Mattos JT, Postma P, Farewell A, Nyström T, Reeh S, Pedersen S, Larsson G (2003) Limiting factors in Escherichia coli fed-batch production of recombinant proteins. Biotechnol Bioeng 81:158–166. https://doi.org/10.1002/bit.10457 25. Scarlat N, Dallemand J-F, Fahl F (2018) Biogas: developments and perspectives in Europe. Renew Energy 129:457–472. https://doi.org/10.1016/j.renene.2018.03.006 26. Smid EJ, Lacroix C (2013) Microbe–microbe interactions in mixed culture food fermentations. Curr Opin Biotechnol 24:148–154. https://doi.org/10.1016/j.copbio.2012.11.007 27. Trad Z, Akimbomi J, Vial C, Larroche C, Taherzadeh MJ, Fontaine J-P (2015) Development of a submerged anaerobic membrane bioreactor for concurrent extraction of volatile fatty acids and biohydrogen production. Bioresour Technol 196:290–300. https://doi.org/10.1016/j.biortech.2015.07.095 28. Tsen S-D (1994) Chemostat selection of Escherichia coli mutants secreting thymidine, cytosine, uracil, guanine, and thymine. Appl Microbiol Biotechnol 41:233–238. https://doi.org/10.1007/BF00186965 28. Tsen S-D (1994) Chemostat selection of Escherichia coli mutants secreting thymidine, cytosine, uracil, guanine, and thymine. Appl Microbiol Biotechnol 41:233–238. https://doi.org/10.1007/BF00186965 29. Wainaina S, Parchami M, Mahboubi A, Horváth IS, Taherzadeh MJ (2019) Food waste-derived volatile fatty acids platform using an immersed membrane bioreactor. Bioresour Technol 274:329– 334. https://doi.org/10.1016/j.biortech.2018.11.104 29. Wainaina S, Parchami M, Mahboubi A, Horváth IS, Taherzadeh MJ (2019) Food waste-derived volatile fatty acids platform using an immersed membrane bioreactor. References Bioresour Technol 274:329– 334. https://doi.org/10.1016/j.biortech.2018.11.104 30. Wolin MJ (1969) Volatile fatty acids and the inhibition of Escherichia coli growth by rumen fluid. Appl Microbiol 17:5 30. Wolin MJ (1969) Volatile fatty acids and the inhibition of Escherichia coli growth by rumen fluid. Appl Microbiol 17:5 Figures Figures Page 15/17 Figure 1 Shake flask cultivation of E. coli W3110 (left) and W3110 ΔFadR (right) on a VFA-rich an food residues supplemented with minimal medium salts (MM) at pH 7.0, 37°C. To assess the strains, the anaerobic digest was supplied in concentration increments between 10-90 with water with constant MM concentration A replicate experiment was performed at 50 Page 16/17 Figure 1 Shake flask cultivation of E. coli W3110 (left) and W3110 ΔFadR (right) on a VFA-rich anaerobic digest of food residues supplemented with minimal medium salts (MM) at pH 7.0, 37°C. To assess the tolerance of the strains, the anaerobic digest was supplied in concentration increments between 10-90 % (v/v) diluted with water, with constant MM concentration. A replicate experiment was performed at 50% (v/v) digest concentration. a) Growth curves for the two strains at various dilutions, supplemented with minimal salts Figure 1 Figure 2 Residual substrate concentrations in aerobic chemostat cultures of W3110 ΔFadR in minimal salts medium with the indicated mix of VFAs (light grey) as carbon source at pH 7.0 and 37°C. The bars show average concentrations with standard deviations (n = 6, biological triplicates each sampled at two different timepoints separated by at least 1/D hours) at two different dilution rates, compared to the initial concentration in the medium. * below the detection limit (10 mg L-1) Figure 1 Shake flask cultivation of E. coli W3110 (left) and W3110 ΔFadR (right) on a VFA-rich anaerobic digest of food residues supplemented with minimal medium salts (MM) at pH 7.0, 37°C. To assess the tolerance of the strains, the anaerobic digest was supplied in concentration increments between 10-90 % (v/v) diluted with water, with constant MM concentration. A replicate experiment was performed at 50% (v/v) digest concentration. a) Growth curves for the two strains at various dilutions, supplemented with minimal salts Page 16/17 Page 16/17 medium (+MM) or MOPS buffer (+MOPS). b and c) Residual acid concentrations in the medium supplemented with 90% anaerobic digest. The concentrations from each separate culture are available in Fig. S2 Figure 2 Residual substrate concentrations in aerobic chemostat cultures of W3110 ΔFadR in minimal salts medium with the indicated mix of VFAs (light grey) as carbon source at pH 7.0 and 37°C. The bars show average concentrations with standard deviations (n = 6, biological triplicates each sampled at two different timepoints separated by at least 1/D hours) at two different dilution rates, compared to the initial concentration in the medium. * below the detection limit (10 mg L-1) medium (+MM) or MOPS buffer (+MOPS). b and c) Residual acid concentrations in the medium supplemented with 90% anaerobic digest. The concentrations from each separate culture are available in Fig. S2 medium (+MM) or MOPS buffer (+MOPS). b and c) Residual acid concentrations in the medium supplemented with 90% anaerobic digest. The concentrations from each separate culture are available in Fig. S2 medium (+MM) or MOPS buffer (+MOPS). b and c) Residual acid concentrations in the medium supplemented with 90% anaerobic digest. The concentrations from each separate culture are available in Fig. S2 Figure 2 Figure 2 20200602VFAPapersupplementary.pdf Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. Page 17/17 Page 17/17
https://openalex.org/W3129342818	https://www.researchsquare.com/article/rs-191703/latest.pdf	English	null	Total Anomalous Pulmonary Venous Connection Requiring Emergency Surgery: What Comes First?	Research Square (Research Square)	2,021	cc-by	5,639	Zhengxia Pan Chongqing Medical University Affiliated Children's Hospital ming xiang (  drxiangming@126.com ) Chongqing Medical University Affiliated Children's Hospital Total Anomalous Pulmonary Venous Connection Requiring Emergency Surgery: What Comes First? Chongqing Medical University Pediatric College: Chongqing Medical University Affiliated Children's Hospital Results The median weight, median age at surgery, mean cardiopulmonary bypass (CPB) duration and mean aortic cross-clamp (ACC) duration were not significantly different between the two groups. The median durations of ventilator support were 8.1±4.6 (2-13) days in the SEO group and 4.9±2.1 (2-18) days in the EO group, resulting in a significant difference (p=0.008). There were 12 operative mortalities (27.3%) in the SEO group and 2 operative mortalities (20%) in the EO group, resulting in no significant difference in mortality (p=0.636). Postoperative complications, such as low cardiac output and arrhythmia, were not significantly different between the two groups. The actuarial survival rates in the EO and SEO groups at 5 years were 87.5% and 89.9%, respectively. There was no difference in actuarial survival between the two groups at the latest follow-up (SEO group 89.9% versus EO group 87.5%, p=0.8115). Background To examine two different methods for treating patients with a total anomalous pulmonary venous connection (TAPVC) who need emergency surgery and to summarize the effects of the two strategies by retrospectively reviewing the data of 54 patients. Research article Research article Keywords: Total anomalous pulmonary venous connection, Surgery, Outcome Posted Date: February 9th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-191703/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at Journal of Cardiothoracic Surgery on August 2nd, 2021. See the published version at https://doi.org/10.1186/s13019-021-01559-y. Page 1/13 Page 1/13 Methods A retrospective review of 54 patients with TAPVC who underwent emergency operations between December 2010 and July 2019 at a single institution was conducted. All patients exhibited respiratory and hemodynamic instability that required mechanical ventilation and inotropic support. Forty-four patients received emergency operations within 24 to 72 hours due to stabilization of the patient’s condition. Stable hemodynamics were achieved, and a constant milieu interne was maintained before the operation. These patients comprised the subemergency operation (SEO) group. Rather than being subjected to efforts to obtain stable hemodynamics and maintain a constant milieu interne, ten patients received emergency operations immediately within 24 hours of diagnosis or an emergency operation is performed immediately due to uncorrectable acidosis or progressive cardiovascular collapse. These patients comprised the emergency operation (EO) group. The hospital course, operative data, and outpatient records were reviewed. Conclusion Rather than making efforts to achieve stable hemodynamics and maintain a constant milieu interne, an emergency operation should be performed immediately, and some efforts to address metabolic acidosis do not reduce mortality. Introduction Page 2/13 Page 2/13 The outcomes of the surgical repair of a total anomalous pulmonary venous connection (TAPVC) have historically improved, with the mortality rates reported in the literature ranging from 10–20%1, 2 or being below 7%3, 4. In contrast, some patients have a risk of progressive pulmonary hypertension (PH) due to pulmonary venous obstruction (PVO), and other patients present with increasing cyanosis, tachypnea, hepatomegaly, hypoxia, gasping, or poor feeding5 due to a delayed diagnosis. All these patients have a risk of progressive PH, suffer from severe respiratory distress and metabolic acidosis and require immediate intubation, nitric oxide and inotropic support; more importantly, an emergency operation is needed. However, there are two viewpoints pertaining to the encounter of this situation. In some patients, the specialist would make efforts to obtain stable hemodynamics, maintain a constant milieu interne, and then perform an emergency operation. This process can take several days. In other patients, an emergency operation is performed immediately instead of trying to obtain stable hemodynamics and maintain a constant milieu interne,what’s more, preoperative medical stabilization is impossible in such cases, even with unstable hemodynamics, an emergency operation is performed. Because of the rarity of reports on these patients who must undergo immediate emergency operations, we conducted a retrospective study to assess the outcomes of such patients. We aimed to determine whether pharmacological interventions to obtain stable hemodynamics and maintain a constant milieu interne are essential. Surgical Technique The surgery was performed using standard cardiopulmonary bypass (CPB). Ice slush was used for myocardial protection in all patients. A temporal reduction in bypass flow could be used to achieve satisfactory intraoperative visualization if needed. The operation was performed through a median sternotomy, there was minimal manipulation of the heart until CPB was established, and the ductus arteriosus was dissected and ligated following the bypass. In patients with supracardiac TAPVC, a long incision was made at the pulmonary venous confluence, and then an incision was made at the corresponding position on the left atrium. Direct side-to-side anastomosis was performed between these two chambers from outside with a running suture to ensure precise geometric alignment and avoid tension, torsion, and rotation; the anastomosis was made to be as large as possible14. This approach was described by Tucker15. The atrial septal defect (ASD) was directly closed with a patch. In patients with cardiac TAPVC, the coronary sinus was unroofed, and a wide tunnel was created between the left atrium and the coronary sinus16. In patients with infracardiac TAPVC, the heart was lifted, and a wide anastomosis was generated between the pulmonary venous chamber and the left atrium with the same procedure as that used for the supracardiac form14. Long-term Follow-up All surgical patients who were discharged alive from the hospital were required to return for outpatient follow-up visits after the initial operation and annually thereafter. Some patients were required to visit local hospitals for routine examinations. Any abnormal examination result or suspicious change in the cardiac condition prompted a return visit for further evaluation. During the follow-up period, echocardiography was performed routinely. Record Review and Definitions In this retrospective analysis, we reviewed the clinical records of patients with isolated TAPVC between December 2010 and July 2019. Patients with single ventricles, atrial isomerism and other complex congenital heart diseases were excluded. Patient characteristics, such as weight and age, pulmonary venous anatomy, demographics, preoperative data, operative data, postoperative data and follow-up data, were recorded. A total of 234 patients who were diagnosed with TAPVC underwent surgical repair at the Children’s Hospital of Chongqing Medical University. Fifty-four patients (23.1%) needed emergency operations for one or more of the following indications: (1) hemodynamic instability, (2) worsening metabolic acidosis, (3) severe progressive cyanosis, or (4) escalating intensive care support as described in other studies6, 7. We performed conventional hyperventilation and pharmacological interventions, such as treatment with prostaglandin E1, milrinone and dopamine, with the aim of controlling PH, maintaining stable hemodynamics and restoring homeostasis5, 8. Forty-four of these patients received emergency operations after their hemodynamics and milieu interne were stabilized, within 24 to 72 hours, and are referred to as the subemergency operation (SEO) group. Additionally, 10 patients received emergency operations within 24 hours after diagnosis due to uncorrectable acidosis or progressive cardiovascular collapse. We refer to these patients as the emergency operation (EO) group. Echocardiography combined with computed tomography angiography (CTA) was performed in all patients in the SEO group. CTA is an accurate, noninvasive, economical diagnostic modality for the Page 3/13 Page 3/13 preoperative evaluation of TAPVC3, 9, 10. Echocardiography was performed in all of the patients in the EO group, and 7 patients underwent CTA. None of the patients in either group underwent cardiac catheterization, given the invasiveness of the procedure. PVO was defined if Doppler examination revealed a pulmonary venous flow velocity > 2 m/s3, 11, 12. Hospital mortality was defined as mortality within 30 days of surgery or before discharge. All other deaths were considered late mortality3, 13. Statistical Analysis Quantitative data are expressed as the mean ± standard deviation as appropriate. All P values were two- sided, and P < 0.05 was considered statistically significant. Age, sex, and type were determined by using the chi-square test. Quantitative data (weight, bypass time, and clamp time) were analyzed by using a t test. Survival data are presented by means of the Kaplan-Meier method. Page 4/13 Page 4/13 Results There were 44 patients in the SEO group, of whom 31 were male and 13 were female. The median weight at surgery was 4.3 ± 1.4 kg (1.7–5.5 kg), and the median age at surgery was 72.4 ± 85.4 days (1-455 days). One patient was 15 months old and lived in a remote mountain area. He had respiratory and hemodynamic instability that required mechanical ventilation. Twenty patients were diagnosed with prerepair PVO; of these patients, 13 exhibited the supracardiac type, 6 exhibited the cardiac type, and 1 exhibited the mixed cardiac type. Ten patients were enrolled in the EO group, of whom 6 were male and 4 were female. The median weight at surgery was 4.3 ± 0.9 kg (2.8–4.9 kg), and the median age at surgery was 37.5 ± 25.5 days (6–90 days). Preoperative PVO occurred in 6 patients; of these patients, 2 exhibited the supracardiac type, 1 exhibited the cardiac type, and 3 exhibited the infracardiac type. Patient demographics are listed in Table 1. Table 1 Demographics and clinical characteristics of the study cohort Demographics SEO (n = 44) EO (n = 10) t/χ2 p Sex Male 31 6 0.413 0.521 Female 13 4 Age at presentation, mean ± SD (d) 72.4 ± 85.4 37.5 ± 25.5 1.273 0.209 Weight at presentation, mean ± SD (kg) 4.3 ± 1.4 4.3 ± 0.9 0.066 0.948 PVO 45.5% (20/44) 60% (6/10) 0.927 0.485 TAPVC type Supracardiac 23 3 4.123 0.248 Cardiac 15 4 Infracardiac 4 3 Mixed 2 0 Table 1 The mean CPB duration was 134.9 ± 42.9 (58–303) minutes in the SEO group and 133.0 ± 41.0 (66–193) minutes in the EO group (p = 0.901). The mean aortic cross-clamp (ACC) duration was 68.2 ± 25.1 (36– The mean CPB duration was 134.9 ± 42.9 (58–303) minutes in the SEO group and 133.0 ± 41.0 (66–193) The mean CPB duration was 134.9 ± 42.9 (58–303) minutes in the SEO group and 133.0 ± 41.0 (66–193) minutes in the EO group (p = 0.901). The mean aortic cross-clamp (ACC) duration was 68.2 ± 25.1 (36– 135) minutes in the SEO group and 54.7 ± 21.5 (range, 27–93) minutes in the EO group (p = 0.123). The median duration of ventilator support was 8.1 ± 4.6 (2–13) days in the SEO group and 4.9 ± 2.1 (2–18) days in the EO group (p = 0.008). The mean CPB duration was 134.9 ± 42.9 (58–303) minutes in the SEO group and 133.0 ± 41.0 (66–193) Results Patient demographics are listed in Table 2. minutes in the EO group (p = 0.901). The mean aortic cross-clamp (ACC) duration was 68.2 ± 25.1 (36– 135) minutes in the SEO group and 54.7 ± 21.5 (range, 27–93) minutes in the EO group (p = 0.123). The median duration of ventilator support was 8.1 ± 4.6 (2–13) days in the SEO group and 4.9 ± 2.1 (2–18) days in the EO group (p = 0.008). Patient demographics are listed in Table 2. Page 5/13 Table 2 Surgical data SEO (n = 44) EO (n = 10) t/χ2 p Mean aortic cross-clamp (ACC) duration (min) 68.2 ± 25.1 54.7 ± 21.5 1.569 0.123 Mean cardiopulmonary bypass duration (min) 134.9 ± 42.9 133.0 ± 41.0 0.125 0.901 Duration of ventilation (d)a 8.1 ± 4.6 4.9 ± 2.1 2.892 0.008 Mortality (n/%) 12 (27.3%) 2 (20%) 0.224 0.636 a: deaths were excluded Table 3 shows the comparison of means and χ2 test results for the complications encountered postoperatively in both groups. Delayed sternal closure was performed in 5 patients and 1 patient in the SEO group and EO group, respectively. Low cardiac output occurred in 9 patients and 2 patients. Arrhythmia occurred in 6 patients and 1 patient. One patient in the SEO group and no patient in the EO group developed PVO. Diaphragmatic paralysis was found in 1 patient in the SEO group and none in the EO group; peritoneal dialysis was performed in 8 patients in the SEO group and 3 patients in the EO group. There were no statistically significant differences between the two groups regarding the complications encountered postoperatively. Table 3 shows the comparison of means and χ2 test results for the complications encountered postoperatively in both groups. Delayed sternal closure was performed in 5 patients and 1 patient in the SEO group and EO group, respectively. Low cardiac output occurred in 9 patients and 2 patients. Arrhythmia occurred in 6 patients and 1 patient. One patient in the SEO group and no patient in the EO group developed PVO. Diaphragmatic paralysis was found in 1 patient in the SEO group and none in the EO group; peritoneal dialysis was performed in 8 patients in the SEO group and 3 patients in the EO group. There were no statistically significant differences between the two groups regarding the complications encountered postoperatively. Results Table 3 Postoperative complications in the two groups SEO (n = 44) EO (n = 10) t/χ2 p Low cardiac output 9 (20.5%) 2 (20%) 0.001 0.974 Peritoneal dialysis 8 (18.2%) 3 (30%) 0.702 0.402 Arrhythmia 6 (13.6%) 1 (10%) 0.095 0.757 Postoperative PVO 1 (2.3%) 0 0.232 0.630 Diaphragmatic paralysis 1(2.3%) 0 0.232 0.630 Delayed sternal closure 5 (11.4%) 1 (10%) 0.015 0.901 Table 3 There were 12 (27.3%) and 2 operative mortalities (20%) in the SEO group and EO group, respectively, but the difference was not significant (p = 0.636). Among the 14 patients who died within 30 days of surgery, the main causes of death were as follows: low output failure (n = 8), pulmonary hypertensive crisis (n = 3), multiple organ dysfunction syndrome (n = 1) and failure to wean from ventilation (n = 1). All patients were weaned off CPB with inotropic support, except for 1 patient in the SEO group. The median follow-up time for survivors was 5.9 years (0.4 to 9.2 years). In the EO group, there was one late mortality of a patient with infracardiac TAPVC 9 months after the operation because of PVO. The Page 6/13 Page 6/13 actuarial survival at 5 years was 87.5% (Fig. 1). In the SEO group, 2 patients were lost to follow-up. Three late deaths occurred after the operation, and all occurred during follow-up at 7 months, 9 months and 1.2 years due to PVO. The actuarial survival at 5 years was 89.9% in the SEO group (Fig. 1). There was no difference in actuarial survival between the two groups at the latest follow-up (SEO group 89.9% versus EO group 87.5%, p = 0.8115). actuarial survival at 5 years was 87.5% (Fig. 1). In the SEO group, 2 patients were lost to follow-up. Three late deaths occurred after the operation, and all occurred during follow-up at 7 months, 9 months and 1.2 years due to PVO. The actuarial survival at 5 years was 89.9% in the SEO group (Fig. 1). There was no difference in actuarial survival between the two groups at the latest follow-up (SEO group 89.9% versus EO group 87.5%, p = 0.8115). Discussion TAPVC is a frequently encountered problem in some heart centers. Some patients have PVO or a delayed diagnosis, and the condition can suddenly worsen, leading to critical illness with respiratory and hemodynamic instability, requiring mechanical ventilation, volume substitution, and inotropic support. Briefly, an emergency operation should be performed in such patients. In this case, someone would make efforts to obtain stable hemodynamics and maintain a constant milieu interne, and then, an emergency operation would be performed. In other cases, an emergency operation would be performed immediately. However, few studies have focused on these patients, and the more appropriate scheme for these patients remains controversial. Treatment of these patients has been a topic of interest and has attracted much attention, but there has been no consensus reached regarding the best strategy. Some reports show that an emergency operation should be performed immediately after a diagnosis is made. Medical efforts are ineffective at managing the hemodynamic and metabolic problems that subsequently occur. Mechanical ventilation, volume substitution and inotropic support do little to help maintain stable hemodynamics and a constant milieu interne. A megadose of vasoactive medications can even worsen cardiac function. Previous studies concluded that immediate emergency surgery was the best strategy, regardless of the high operative risk5, 17, 18. In contrast, stabilizing the patient’s condition before surgery is considered important for improving the surgical outcome. Some studies have reported the use of vasoactive medications such as dopamine, prostaglandin E1 and milrinone to stabilize hemodynamics and correct metabolic acidosis, and when the patient’s condition was stable, an urgent operation was performed. Although one study reported a reduction in mortality, there was insufficient evidence to support this conclusion17. In addition, other palliative surgical techniques, such as balloon dilatation, stenting and atrial septectomy, are alternative approaches to relieve cyanosis and improve the patient’s clinical state. These palliative techniques have been shown to be somewhat successful for decompressing pulmonary venous pressure with restricted ASD or PVO. These techniques are associated with some inherent complications, such as the need for reintervention (due to occlusion of the stent or the recurrence of PVO) and the need for anticoagulation. Research has shown that the mortality rate is 38% at 1 year and that the reintervention rate is 58% at 1 year2, 5, 19. Moreover, not all patients could tolerate these palliative measures, and patients needed to undergo surgery to correct the anomalous pulmonary veins5. Discussion Extracorporeal membrane oxygenation In contrast, stabilizing the patient’s condition before surgery is considered important for improving the surgical outcome. Some studies have reported the use of vasoactive medications such as dopamine, prostaglandin E1 and milrinone to stabilize hemodynamics and correct metabolic acidosis, and when the patient’s condition was stable, an urgent operation was performed. Although one study reported a reduction in mortality, there was insufficient evidence to support this conclusion17. In addition, other palliative surgical techniques, such as balloon dilatation, stenting and atrial septectomy, are alternative approaches to relieve cyanosis and improve the patient’s clinical state. These palliative techniques have been shown to be somewhat successful for decompressing pulmonary venous pressure with restricted ASD or PVO. These techniques are associated with some inherent complications, such as the need for reintervention (due to occlusion of the stent or the recurrence of PVO) and the need for anticoagulation. Research has shown that the mortality rate is 38% at 1 year and that the reintervention rate is 58% at 1 year2, 5, 19. Moreover, not all patients could tolerate these palliative measures, and patients needed to undergo surgery to correct the anomalous pulmonary veins5. Extracorporeal membrane oxygenation Page 7/13 Page 7/13 (ECMO) is also an alternative approach, but its cost and serious complications restrict its popularization2, 20. 20. A finding of our review is that the mortality rate was higher in the SEO group than in the EO group, though without significance. The reasons might be as follows. First, according to some clinical reports, there is evidence that the long-term application of cardiovascular drugs does not improve myocardial function or survival21–23. Indeed, some researchers have shown that the use of these drugs results in myocardial injury, the destruction of myocardial function via myocardial cellular apoptosis and a decrease in heart contractility, which substantially affects myocardial function21. Second, due to severe dyspnea and aggravated metabolic acidosis, these patients’ hearts contracted weakly, and their oxygenation continued to worsen unless the malformation was corrected. Additionally, their cardiac function worsened during the preoperative preparation period1. Another more important and reasonable finding is that the duration of ventilation was significantly longer in the SEO group. Longer postoperative mechanical ventilation results in a higher risk of ventilator- associated pneumonia and prolongs the length of stay. Additionally, studies have shown that prolonged postoperative mechanical ventilation is the most significant predictor of early mortality24. Discussion Long postoperative mechanical ventilation is usually attributed to the presence of chest infection or a worsened condition of heart failure. Therefore, the longer postoperative mechanical ventilation period in the SEO group might be explained as follows. First, an infusion of excess fluid, aiming at stabilizing hemodynamics and correcting metabolic acidosis before the operation, has been demonstrated to worsen mechanical ventilation-related complications and aggravate pneumonia in critically ill patients6, 24. Second, an infusion of excess fluid and long-term application of cardiovascular drugs can also weaken myocardial functioning and prolong mechanical ventilation, as discussed above. Our report indicates that early mortality among these critically ill patients (in both the SEO group and the EO group) is higher than the overall TAPVC mortality reported by others. It seems that those who undergo emergency operations have a higher overall mortality rate2. Clearly, patients with respiratory distress and metabolic acidosis or PVO are the most severely ill patients, and these factors have been shown to be risk factors for mortality in many studies. The urgency of surgery itself is also a risk factor for mortality1, and therefore, such patients are expected to have a higher mortality rate1, 4, 11. Moreover, in some patients, the hemodynamic status was still unstable after all of the measures mentioned above had been implemented, and an emergency operation became the only choice when the palliative techniques were abandoned, considering the aforementioned defects. Regarding the emergency operations themselves, the early mortality is similar to the result of other reports1, 3, 24. Postoperative complications, such as low cardiac output and arrhythmia, were not significantly different between the two groups. Long-term survival also did not differ between the two groups. Operative survivors in the two groups (among critically ill patients) appear to have similar long-term outcomes, such as overall actuarial survival. Zhao K et al. also reported a similar result1. Postoperative patients may have Page 8/13 Page 8/13 late pulmonary vein obstruction, as seen in other patients with TAPVC; the proportion of patients is similar to that with overall TAPVC postoperative PVO3. In other words, patients undergoing an emergency operation are unlikely to be more predisposed to stenosis. The statistical analyses in our series showed a good outcome, with freedom from medication and normal activity. late pulmonary vein obstruction, as seen in other patients with TAPVC; the proportion of patients is similar to that with overall TAPVC postoperative PVO3. Discussion In other words, patients undergoing an emergency operation are unlikely to be more predisposed to stenosis. The statistical analyses in our series showed a good outcome, with freedom from medication and normal activity. Considerable research has been performed to identify the risk factors for mortality among patients with TAPVC, while little research has been devoted to the subset of patients undergoing emergency surgery. Our study showed that the mortality rate was higher in the SEO group than in the EO group, although without significance. Moreover, the duration of ventilation was significantly longer in the SEO group than in the EO group, which was not previously reported. Therefore, emergency operations should be performed immediately to correct cardiac malformations rather than taking efforts to obtain stable hemodynamics and maintain a constant milieu interne. Medical treatments and other processes do not reduce mortality. There are multiple limitations of this study. This was a retrospective single-center study, and the sample size was small; thus, the results may not be applicable to the population as a whole. TAPVC often combined with various malformations. Although cases combined with malformations of high risk for death were excluded, other concurrent abnormalities could still affect survival outcomes. Conclusion Taken together, our experience with these critically ill patients is as follows. First, given the lower mortality rate (though without significance compared to the SEO group) and shorter duration of ventilation in the EO group, emergency operations should be performed without any delay. The evaluation of the clinical state before the operation should be accurate and rapid, and focus should not be placed on maintaining stable hemodynamics and a constant milieu interne. Second, for patients with respiratory and hemodynamic instability, mechanical ventilation and inotropic support should be applied quickly. Abbreviations TAPVC: total anomalous pulmonary venous connection; PVO: pulmonary venous obstruction; subemergency operation (SEO); emergency operation (EO); cardiopulmonary bypass (CPB) duration and mean aortic cross-clamp (ACC); pulmonary hypertension (PH) because of pulmonary venous obstruction (PVO); computed tomography angiography (CTA); atrial septal defect (ASD); extracorporeal membrane oxygenation (ECMO) Declarations Ethics approval and consent to participate: This research received ethical approval and consent. It did not require the approval of an ethics committee. Page 9/13 Page 9/13 Consent for publication: All patient information contained in this manuscript has been published with the consent of their parents. Availability of data and materials: The data and materials in the manuscript are available, and the original data for the relevant results are owned by myself; I can be contacted if needed. Competing interests: There are no conflicts of interest or legal economic disputes in this study. Funding: Not applicable. Funding: Not applicable. Authors' contributions: Linyun Xi, the first author, designed the work, acquired, analyzed, and interpreted the data, drafted the work and substantively revised it. Chun Wu, the second author, substantively revised it. Zhengxia Pan, the third author, provided some suggestions for this work. Ming Xiang, the corresponding author, designed the work, drafted the work, substantively revised it, and approved the submitted version. Acknowledgements: I would like to thank all the doctors and nurses of the Cardiothoracic Surgery Department of the Children’s Hospital of Chongqing Medical University for giving me so much advice and help. I appreciate my family for their companionship, support and care. Acknowledgements: I would like to thank all the doctors and nurses of the Cardiothoracic Surgery Department of the Children’s Hospital of Chongqing Medical University for giving me so much advice and References 1. Zhao K, Wang H, Wang Z, Zhu H, Fang M, Zhu X, Zhang N and Song H. Early-and intermediate-term results of surgical correction in 122 patients with total anomalous pulmonary venous connection and biventricular physiology. Journal of cardiothoracic surgery. 2015;10:172. 1. Zhao K, Wang H, Wang Z, Zhu H, Fang M, Zhu X, Zhang N and Song H. Early-and intermediate-term results of surgical correction in 122 patients with total anomalous pulmonary venous connection and biventricular physiology. 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https://openalex.org/W3111399829	https://hal.archives-ouvertes.fr/hal-03127528/file/v11p1207.pdf	English	null	Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): a systematic review of <i>in vitro</i>, <i>in vivo</i>, and clinical trials	Theranostics	2,021	cc-by	25,210	To cite this version: Youngjoo Han, Keum-Hwa Lee, Sojung Yoon, Seoung-Wan Nam, Seohyun Ryu, et al.. Treatment of se- vere acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): A systematic review of in vitro, in vivo, and clinical trials. Theranostics, 2021, 11 (3), pp.1207-1231. 10.7150/thno.48342. hal-03127528 Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): A systematic review of in vitro, in vivo, and clinical trials Youngjoo Han, Keum-Hwa Lee, Sojung Yoon, Seoung-Wan Nam, Seohyun Ryu, Dawon Seong, Jae-Seok Kim, Jun-Young Lee, Jaewon Yang, Jinhee Lee, et al. To cite this version: Youngjoo Han, Keum-Hwa Lee, Sojung Yoon, Seoung-Wan Nam, Seohyun Ryu, et al.. Treatment of se- vere acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): A systematic review of in vitro, in vivo, and clinical trials. Theranostics, 2021, 11 (3), pp.1207-1231. 10.7150/thno.48342. hal-03127528 HAL Id: hal-03127528 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. 1207 Theranostics 2021, Vol. 11, Issue 3 Ivyspring International Publisher Ivyspring International Publisher Received: 2020.05.18; Accepted: 2020.10.22; Published: 2021.01.01 Received: 2020.05.18; Accepted: 2020.10.22; Published: 2021.01.01 Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): a systematic review of in vitro, in vivo, and clinical trials Department of Psychosis Studies, Institute of Psychiatry, Psychology and Neuroscience, King’s Coll p y y y y gy g g 13. Centre for Psychiatric Research, Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden. p y y y y gy g g 13. Centre for Psychiatric Research, Department of Clinical Neuroscience, Karolinska Institutet, Stockholm p y y y y gy entre for Psychiatric Research, Department of Clinical Neuroscience, Karoli y p 14. The Cambridge Centre for Sport and Exercise Sciences, Anglia Ruskin University, Cambridge, UK. y 14. The Cambridge Centre for Sport and Exercise Sciences, Anglia Ruskin 15. School of Social Work, University of Southern California, CA, USA. y 16. Division of Urology, Brigham and Women’s Hospital and Harvard Medical School, Bosto y 16. Division of Urology, Brigham and Women’s Hospital and Harvard M 16. Division of Urology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA. gy, g p , , , 7. Department of Internal Medicine IV (Nephrology and Hypertension), Medical University Innsbruck, Innsbruck, Austria. gy g p 17. Department of Internal Medicine IV (Nephrology and Hypertension), Medical University Innsbruck, Innsbruck, gy g p 17. Department of Internal Medicine IV (Nephrology and Hypertension), 9. Department of Internal Medicine, St. Johann County Hospital, St. Johann in Tirol, Austria. 19. Department of Internal Medicine, St. Johann County Hospital, St. Johann in Tirol, Austria. 20. Department of Internal Medicine, University of Illinois College of Medicine at Peoria, Peoria, IL, USA. . Department of Internal Medicine, University of Illinois College of Medic p , y g , , , 21. Faculty of Medicine, University of Versailles Saint-Quentin-en-Yvelines, Montigny-le-Bretonneux, France. p y g 1. Faculty of Medicine, University of Versailles Saint-Quentin-en-Yvelines, Montigny-le-Bretonneux, France. 21. Faculty of Medicine, University of Versailles Saint-Quentin-en-Yvelines, Montigny-le-Bretonneux, France  Corresponding author: Dr. Jae Il Shin MD PhD, 50-1 Yonsei-ro, Seodaemun-gu, Department of Pediatrics, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. Tel: 82-2-2228-2050, Fax: 82-2-393-9118, E-mail: shinji@yuhs.ac.  Corresponding author: Dr. Jae Il Shin MD PhD, 50-1 Yonsei-ro, Seodaemun-gu, Department of Pediatrics, Yonsei University College of Medicine, Seoul 03722, Republic of Korea. Tel: 82-2-2228-2050, Fax: 82-2-393-9118, E-mail: shinji@yuhs.ac.  Corresponding author: Dr. Jae Il Shin MD PhD, 50-1 Yonsei-ro, Seodaemun-gu, Department of Pediatrics, Yonsei University College of M Korea. Tel: 82-2-2228-2050, Fax: 82-2-393-9118, E-mail: shinji@yuhs.ac. © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): a systematic review of in vitro, in vivo, and clinical trials See http://ivyspring.com/terms for full terms and conditions. © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativec ee http://ivyspring.com/terms for full terms and conditions. Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): a systematic review of in vitro, in vivo, and clinical trials Treatment of severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus disease 2019 (COVID-19): a systematic review of in vitro, in vivo, and clinical trials Young Joo Han1, Keum Hwa Lee2, Sojung Yoon3, Seoung Wan Nam4, Seohyun Ryu3, Dawon Seong3, Jae Seok Kim5, Jun Young Lee5, Jae Won Yang5, Jinhee Lee6, Ai Koyanagi7,8, Sung Hwi Hong9, Elena Dragioti10, Joaquim Radua11,12,13, Lee Smith14, Hans Oh15, Ramy Abou Ghayda9,16, Andreas Kronbichler17, Maria Effenberger18, Daniela Kresse19, Sara Denicolò17, Woosun Kang20, Louis Jacob7,21, Hanwul Shin5, and Jae Il Shin2 1. Department of Pediatrics, Samsung Changwon Hospital, Sungkyunkwan University School of Medicine, Changwon, Republic of Korea. 2. Department of Pediatrics, Yonsei University College of Medicine, Seoul, Republic of Korea. p y g 3. Yonsei University College of Medicine, Seoul, Republic of Korea. p gy y j g j p . Department of Nephrology, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea. p gy y j g j p 5. Department of Nephrology, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea. 6. Department of Psychiatry, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea. 7 Research and development unit Parc Sanitari Sant Joan de Déu/CIBERSAM Universitat de Barcelona F 6. Department of Psychiatry, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea. ́ ́ ́ 7. Research and development unit, Parc Sanitari Sant Joan de Déu/CIBERSAM, Universitat de Barcelona, Fundació Sant Joan de Déu, San 8. ICREA, Pg. Lluis Companys 23, 08010, Barcelona, Spain. 8. ICREA, Pg. Lluis Companys 23, 08010, Barcelona, Spain. g p y p 9. Department of Global Health and Population, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, USA. 9. Department of Global Health and Population, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, USA. 10. Pain and Rehabilitation Centre, and Department of Health, Medicine and Caring Sciences, Linköping University, Linköping, Sweden. 9. Department of Global Health and Population, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, USA. 10. Pain and Rehabilitation Centre, and Department of Health, Medicine and Caring Sciences, Linköping University, Linköping, Sweden g q g y ( ) g ( ), 12. Department of Psychosis Studies, Institute of Psychiatry, Psychology and Neuroscience, King’s College London, London, UK. g q g y ( ) 12. Introduction Coronavirus disease 2019 (COVID-19) refers to a respiratory syndrome caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS- CoV-2), an RNA virus belonging to the Coronaviridae family. Ever since the disease was first reported in Wuhan, China in December 2019, it has spread rapidly around the world. On October 28, 2020, a total of 43,766,712 SARS-CoV-2 cases were reported worldwide, of which 1,163,459 died [1]. Clinical manifestations range from being asymptomatic to pneumonia and acute respiratory distress syndrome (ARDS). Although estimations of case-fatality rate are different for COVID-19, there appears to be a high rate of a severe disease course or death, mainly in patients with advanced age or underlying diseases [2, 3]. Current case fatality rates are 2.2% in Africa, 3.2% in Americas, 2.5% in Eastern Mediterranean Region, 3.2% in Europe, 1.6% in South-East Asia, and 2.1% in Western Pacific Region [1], whereas the case fatality rate of SARS and Middle East respiratory syndrome (MERS), which are coronavirus respiratory syndromes similar to COVID-19 were 11% [4] and 34% [5], respectively. q )] ( g ) Moreover, in order to search for studies on COVID-19, a search was performed through the following search algorithm until the last update on May 7, 2020: ((wuhan[All Fields] and (“corona- virus”[MeSH Terms] or “coronavirus”[All Fields])) and 2019/12[PDAT]: 2030[PDAT]) or 2019-nCoV[All Fields] or 2019nCoV[All Fields] or COVID-19[All Fields] or SARS-CoV-2[All Fields]. In addition, a search for randomized controlled trials (RCTs) on COVID-19 was also performed using the following search terms until the last update on October 9, 2020: ((((wuhan[All Fields] and (“coronavirus”[MeSH Terms] or “coronavirus” [All Fields])) and 2019/12 [PDAT]: 2030[PDAT]) or 2019-nCoV[All Fields] or 2019nCoV [All Fields] or COVID-19[All Fields] or SARS-CoV-2[All Fields]) and (random [Title/Abstract] or randomization [Title/Abstract] or randomized [Title/Abstract] or randomized [Title/Abstract] or trial[Title]). To include a more sufficient amount of RCTs, a search for preprint RCTs through the database of Medrxiv.org was performed by conditions that include the following search terms in the titles until the last update on October 11, 2020: [“COVID” and (“random”, “controlled”, or “trial”)] or [“coronavirus” and (“random”, “controlled”, or “trial”)]or [“cov” and (“random”, “controlled”, or “trial”)]. Abstract Rationale: Coronavirus disease 2019 (COVID-19) has spread worldwide and poses a threat to humanity. However, no specific therapy has been established for this disease yet. We conducted a systematic review to highlight therapeutic agents that might be effective in treating COVID-19. Methods: We searched Medline, Medrxiv.org, and reference lists of relevant publications to identify articles of in vitro, in vivo, and clinical studies on treatments for severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and COVID-19 published in English until the last update on October 11, 2020. Results: We included 36 studies on SARS, 30 studies on MERS, and 10 meta-analyses on SARS and MERS in this study. Through 12,200 title and 830 full-text screenings for COVID-19, eight in vitro studies, 46 randomized controlled trials (RCTs) on 6,886 patients, and 29 meta-analyses were obtained and investigated. There was no therapeutic agent that consistently resulted in positive outcomes across SARS, MERS, and COVID-19. Remdesivir showed a therapeutic effect for COVID-19 in two RCTs involving the largest number of total participants (n = 1,461). Other therapies that showed an effect in at least two RCTs for COVID-19 were sofosbuvir/daclatasvir (n = 114), colchicine (n = 140), IFN-β1b (n = 193), and convalescent plasma therapy (n = 126). Conclusions: This review provides information to help establish treatment and research directions fo COVID-19 based on currently available evidence. Further RCTs are required. Key words: COVID-19, therapeutic agent, SARS, MERS, mortality, coronavirus http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1208 manner during this pandemic, we first searched the meta-analyses or systematic reviews on SARS and MERS from inception to March 31, 2020 using the following search terms (“severe acute respiratory syndrome”, “SARS”, “Middle East respiratory syndrome”, or “MERS”) and (“meta”[title] or “systematic” [title]). After reading the full-text of articles obtained as a result of this search, we also investigated the in vitro, in vivo, and human studies on therapeutics of SARS or MERS that were included in them. Next, we conducted an additional search using the following search terms for the parts that were considered to be necessary for replenishment: [(“severe acute respiratory syndrome” or “SARS”) and (“remdesivir”, “nelfinavir”, “interferon beta”, or “chloroquine”)] or [(“Middle East respiratory syndrome” or “MERS”) and (“remdesivir”, “lopinavir”, “ritonavir”, “interferon alpha”, “interferon beta”, “convalescent plasma”, “chloroquine”, or “corticosteroid”)] (Figure 1). Introduction A search for meta-analyses of treatment for COVID-19 was performed using the following search terms until the last update on October 11, 2020: ((((wuhan[All Fields] and (“coronavirus”[MeSH Terms] or “coronavirus”[All Fields])) and 2019/12[PDAT]: 2030[PDAT]) or 2019-nCoV[All Fields] or 2019nCoV[All Fields] or COVID-19[All Fields] or SARS-CoV-2[All Fields]) and (meta[Title]) (Figure 2). There are currently no specific established treatments for COVID-19. Since the outbreak of COVID-19, numerous studies have been conducted during the past months; however, it is difficult to extract information from these extensive studies, synthesize the results, and apply them in practice. In fact, it would be almost impossible for front-line medical practitioners to be able to absorb the considerable number of reports being released on a daily basis and immediately translate the findings into practice during this medical crisis. For this reason, we summarized the in vivo, in vitro, and clinical research results related to potential therapies of COVID-19 and further integrated the results with previously reported results from SARS and MERS. We aimed to provide useful information for the establishment of treatment and research directions for COVID-19. Literature search strategy and study selection We adhered to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. Two investigators (YJH and JIS) manually searched Medline for literature regarding therapeutics for SARS, MERS, and COVID-19. Only publications in English were included, with the exception of an individual study used within a meta-analysis. In order to complete this review in a timely http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1209 Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East respiratory syndrome; RCT: randomized controlled trial. Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with in vitro studies on MERS. Theranostics 2021, Vol. 11, Issue 3 1209 Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East respiratory syndrome; RCT: randomized controlled trial Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with in vitro studies on MERS 1209 Theranostics 2021, Vol. 11, Issue 3 Theranostics 2021, Vol. 11, Issue 3 1209 Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East respiratory syndrome; RCT: randomized controlled trial. Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with in vitro studies on MERS. Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East respiratory syndrome; RCT: randomized controlled trial. Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with in vitro studies on MERS. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East resp d controlled trial. Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle andomized controlled trial Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlap Figure 1. Flowchart of article selection process for SARS and MERS. SARS: severe acute respiratory syndrome; MERS: Middle East respiratory syndrome; RCT: randomized controlled trial. Literature search strategy and study selection Two overlapped with in vitro studies on SARS; †One overlapped with an in vitro study on SARS each; ‡One overlapped with in vitro studies on MERS. Figure 2. Flowchart of article selection process for COVID-19. COVID-19: coronavirus disease 2019; PCR: polymerase chain reaction; RCT: randomized controlled trial. Including non-RCTs on corticosteroid therapy for patients with various severity of COVID-19. †Studies on angiotensin-converting enzyme inhibitor or angiotensin receptor blocker. e selection process for COVID-19. COVID-19: coronavirus disease 2019; PCR: polymerase chain reaction; RCT: randomized controlled rticosteroid therapy for patients with various severity of COVID-19. †Studies on angiotensin-converting enzyme inhibitor or angiotensin Figure 2. Flowchart of article selection process for COVID-19. COVID-19: coronavirus disease 2019; PCR: polymerase chain reaction; RCT: randomized controlled trial. Including non-RCTs on corticosteroid therapy for patients with various severity of COVID-19. †Studies on angiotensin-converting enzyme inhibitor or angiotensin receptor blocker. were (1) studies on SARS-CoV, MERS-CoV, or SARS-CoV-2 and (2) studies in which inoculation of virus preceded administration of therapeutic agents. For human research, eligibility criteria were organized in accordance with the Participants, Interventions, Comparisons, and Outcomes (PICO) reporting structure. Data extraction Two investigators (YJH and JIS) collected information on the total number of patients and the number of patients in the intervention group, time range of enrollment or at the time of diagnosis or hospitalization, intervention and control therapy used in the study, and the outcome among the intervention and the control group. Eligibility criteria Two investigators (YJH and JIS) identified the eligible studies by screening the titles and abstracts independently. Any disagreement was resolved by discussion and consensus among review authors. For non-human research, eligibility criteria for inclusion http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1210 Classification of studies and interpretation of results In order to interpret the results of in vitro studies, 50% maximal effective concentration (EC50) less than 10 μM or selectivity index (SI) greater than 10 was set as a criterion for determining whether a particular drug has therapeutic potential against the virus of interest. Comparisons Control interventions relevant to the general treatment of respiratory infection (e.g., placebo or usual medications) or other therapeutic agents that could be candidates for the study intervention were included. Not effective: The results for any outcome did not show a significant difference between the treatment and the control group (P > 0.05). Possible harm: The treatment group did not show statistically superior results for major outcomes (P > 0.05), and the results for other outcomes were worse with a statistical significance (P < 0.05). Interventions We considered the pharmacological, immuno- logical, or miscellaneous therapies administered after the onset of infection. Multiple therapeutic agents in combination were also included. Types of respiratory support, mechanical ventilation (MV) strategy, extracorporeal therapy, and radiation therapy were not target interventions in this study. The exclusion criteria were studies on (1) immunization or chemoprophylaxis, (2) Chinese medicine, or (3) other topics, such as epidemiology, without dealing with therapeutic interventions. We also excluded non-RCTs that did not specify the number of patients in the intervention group. The results of RCTs and meta-analyses were categorized as follows, depending on whether the therapeutic agent was effective against COVID-19. Effective: The treatment group showed superior results for major outcomes (mortality, ICU admission, disease progression, discharge, clinical improvement, or improvement in the chest radiograph) with a statistical significance (P < 0.05). Possible effect: The major outcome of the treatment group was not significantly worse (P > 0.05), and the results for other outcomes other than the major outcome was superior in the treatment group with a statistical significance (P < 0.05). Study design We included studies on individuals with SARS, MERS, or COVID-19 who were diagnosed by validated methods using real time reverse transcription polymerase chain reaction (PCR) [6]. We excluded studies that were performed exclusively in children. According to the 7th edition of the Chinese clinical guidance for COVID-19 pneumonia, treatment with corticosteroids, tocilizumab, or convalescent plasma was recommended for patients with severe or progressive COVID-19 [7]. Therefore, when a non-RCT was included in a meta-analysis and targeted any of these treatment forms for patients with different severity of COVID-19, only the study analyzing the results of multivariate analysis conducted in the original research was included. In the case of meta-analyses on angiotensin-converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) for COVID-19, studies that did not include participants selectively according to the presence of hypertension were excluded because it was thought that a mixture of participants with and without hypertension would affect the treatment outcome. Because RCTs on SARS or MERS performed to date were not sufficient, any RCT, study in prospective or retrospective cohort design, case- control design, or case series published as an article in a scientific journal were eligible. In the case of non-RCTs, studies with a total of 10 or more patients were included, except for relatively rare treatment forms that had not been administered in dozens of patients to date. For COVID-19, only RCTs were eligible except of studies included in a meta-analysis. Systematic search results Through Medline search, a total of 10 meta-analyses on SARS (n = 5) [8-12], MERS (n = 3) [13-15], and both (n = 2) [16, 17] were obtained and investigated. After investigating the original texts of in vitro, in vivo, and clinical studies cited in these meta-analyses, an additional Medline search was performed when the clinical study obtained by the search seemed to be insufficient for therapeutic agents that showed positive results from in vitro or in vivo studies. Through this process, 36 and 30 eligible articles on SARS and MERS were obtained, respectively: 20 in vitro, five in vivo studies (two overlapping with in vitro studies on SARS), 13 human non-RCTs (one overlapping with an in vitro study on SARS), and one RCT on SARS; and 15 in vitro (one overlapping with an in vitro study on SARS), seven in vivo studies (one overlapping with in vitro studies on MERS), and 10 human non-RCTs on MERS. In addition, as a result of searching the database of Clinicaltrials.gov, we identified one RCT on SARS that was not completed after registration, and one on MERS. Of these, an RCT of lopinavir/ritonavir plus ribavirin in the treatment of SARS [18] had not yet started recruiting participants since it was registered in December 2007, and the current status was unknown; and another RCT of lopinavir/ritonavir and interferon (IFN)-β1b in the treatment of MERS [19] was completed on May 20, 2020 (Figure 1). ( ) [ ] ( ) The other two RCTs for COVID-19 were performed with different administration periods of remdesivir, five and ten days, respectively. Among them, the 5-day treatment group showed better clinical status distribution on the 7-category ordinal scale on day 11 (OR 1.65, 95% CI 1.09 to 2.48) in one RCT for moderate COVID-19 [23]. In another RCT for severe COVID-19, the incidence of serious adverse events was lower in the 5-day treatment group than in the 10-day treatment group (42 out of 200 [21%] vs. 68 out of 197 [35%]; difference 10.8%, 95% CI 2.4% to 19.2%) [24] (Table 3). In a meta-analysis involving one of these RCTs [24] and another unreported RCT [25], the OR for clinical recovery in the 5-day course of treatment was 1.33 (95% CI 1.01 to 1.76) compared to the 10-day course of treatment [26] (Table 4). A total of 12,200 articles on COVID-19 were identified through a Medline and Medrxiv.org search. Outcomes Studies reporting on mortality, intensive care unit (ICU) admission, disease progression, discharge rates, or improvement in the chest radiograph in the intervention/entire patient group or control group were included. Harmful: The treatment group showed statistically inferior results for the major outcome (P < 0.05). http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1211 (RR) for recovery 1.32, 95% CI 1.12 to 1.55; P < 0.0001} and the odds ratio (OR) for the improvement of the ordinal score on day 15 was 1.50 (95% CI 1.18 to 1.91, P = 0.001). There was no significant difference in the 14-day mortality rate between the two groups. However, when compared among the participants with a baseline ordinal score of 5 requiring oxygen supplementation, the 14-day mortality rate of the treatment group was significantly lower (4 out of 222 [2%] vs. 19 out of 199 [10%]; hazard ratio [HR] 0.22, 95% CI 0.08 to 0.58) [20]. In another RCT on severe COVID-19, the 28-day mortality rate did not differ between remdesivir-treated patients and controls [21] (Table 3). According to a meta-analysis for these two RCTs [20, 21], the RR for clinical recovery was 1.17 (95% CI 1.07 to 1.29) [22] (Table 4). Systematic search results After full-text screening of 830 articles, 83 eligible articles on COVID-19 were obtained: eight in vitro studies, 46 RCTs on 6,886 patients, and 29 meta-analyses (Figure 2). Sofosbuvir and daclatasvir A combination of sofosbuvir/daclatasvir showed an effect in two RCTs on COVID-19 which were conducted in Iran. In one RCT, the cumulative incidence of hospital discharge was higher (P = 0.041) and the duration of hospitalization was shorter (6 [interquartile range (IQR) 4–8] vs. 8 days [5–13]; P = 0.029) in the treatment compared to the control group [27]. In another RCT, the cumulative incidence of recovery was higher in the treatment compared to the control group (P = 0.033) [28] (Table 3). The research results for SARS and MERS for each therapeutic agent are described in Table 1, and the research results for COVID-19 are described in Table 2, 3 & Table 4, and Table S1. Favipiravir Remdesivir showed effects in multiple non-human studies on SARS or MERS (Table 1), and in one in vitro study on COVID-19 (Table 2). Four RCTs on remdesivir for COVID-19 have been published to date. One of them was a large-scale RCT, with 538 and 521 participants in the treatment and the control group, respectively, and remdesivir was administered to the treatment group for 10 days. The time to recovery was shorter in the treatment group compared to the control group {11 [95% confidence interval (CI) 9–12] vs. 15 [13–19] days; relative risk The results of two in vitro studies on favipiravir for COVID-19 were unfavorable [29, 30]. However, in a Russian RCT on favipiravir for moderate COVID-19, the rate of negative results of virus PCR on day 5 was higher in the treatment than in the control group (25 out of 40 [63%] vs. 6 out of 20 [30%]; P = 0.018) [31]. In another RCT on mild COVID-19, the hospital discharge rate of participants who received favipiravir from the first day of enrollment was higher http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1212 than that of participants who received favipiravir starting from one week after enrollment (HR 2.68, 95% CI 1.67 to 4.29) [32] (Table 3). treatment with umifenovir did not show better outcome than the control group in terms of clinical deterioration or viral clearance [34] (Table 3). On the other hand, a meta-analysis that included this RCT [34] and four observational studies on COVID-19 demonstrated that umifenovir treatment enhanced the rate of viral clearance on day 14 (RR 1.27, 95% CI 1.04 to 1.55; P = 0.02; I2 = 63%; n = 683) [35] (Table 4). Umifenovir Umifenovir showed an effect in an in vitro study on COVID-19 [33] (Table 2). In an RCT comparing a combination of umifenovir and lopinavir/ritonavir with standard treatment, the group receiving the Table 1. Summary of studies evaluating therapeutics for SARS and MERS Therapeutics SARS MERS In vitro In vivo Human In vitro In vivo Human Antiviral agents Ribavirin 4 studies [36, 130-132] 2 studies [133, 134] 4 studies [135-138] 1 study [139] Remdesivir 1 study [140] 1 study [140] 4 studies [38, 140-142] 2 studies [38, 143] Lopinavir 1 study [130] 2 studies [48, 50] 2 studies [38, 51] 1 study [95] Ritonavir 1 study [48] Oseltamivir 1 study [136] 1 study [144] Nelfinavir 1 study [48] 1 study [50] 1 study [50] Interferon IFN-α (8 studies) [50, 130, 132, 134, 145-148]; IFN-β (8 studies) [130, 132, 145, 147-151] IFN-α/IL-1β (1 study) [152] IFN-α B/D, rintatolimod† (1 study) [50]. IFN-α (1 study): more effective than corticosteroids [153]. IFN-α (1 study) [139]; IFN-β (2 studies) [154, 155]. IFN-β (2 studies) [39, 156]§ 1 study [14]‡ IFN-α-n3 (1 study) [50] Combination therapy based on antiviral agents or interferon Ribavirin/IFN IFN-α (1 study) [130]; IFN-β (2 studies) [130, 131]. IFN-α (1 study) [157] IFN-α (2 studies) CFR 6/20 (30%) vs. 17/24 (71%) (P = 0 01) [158] Table 1. Summary of studies evaluating therapeutics for SARS and MERS IFN-α (2 studies) CFR 6/20 (30%) vs. 17/24 (71%) (P = 0.01) [158] CFR 14/61 (23%) vs. 2/2 (100%) (P = 0.01) [159]. 4 studies [144, 160-162]: no difference in mortality. difference in mortality. Ribavirin/ lopinavir 1 study [36] Ribavirin plus L/r Registered RCT (not yet recruiting) [18] L/r 3 studies [36, 99, 130] 1 study [48] 2 studies: Rates of ARDS/death (2% vs. 29%, P = 0.001) [36] CFR 2% vs. 16% (P < 0.05) [37]. Umifenovir 1 study [38] 1 study [39] L/r plus IFN-β 2 studies [38, 39] Ongoing RCT [19] Ribavirin/ corticosteroids 1 study earlier administration [163] IFN-α/ corticosteroids 1 study [153] IFN-β/IFN-γ 1 study [164] Intranasal IFN-β/ HR2P-M2 1 study [156] Antibiotics Macrolide 1 study: mortality and viral clearance [165] 4-Aminoquinoline Chloroquine 3 studies [50, 166, 167] 1 study [50] 1 study [51] 1 study [52] Amodiaquine 1 study [50] 1 study [50] Corticosteroids 4 studies Higher dose [62, 63] High dose methylprednisolone [64] Methylprednisolone Inconclusive (2 studies); Delay in viral clearance (HR 0.35; 95% CI 0.17-0.72), not associated with 1 study: mortality and viral clearance [165] Inconclusive (2 studies); Delay in viral clearance (HR 0.35; 95% CI 0.17-0.72), not associated with http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1213 Therapeutics SARS MERS In vitro In vivo Human In vitro In vivo Human was better than 3 other groups (no steroid, hydrocortisone, or pulse therapy) [65]. mortality [61] CFR 6/13 (46%) vs. 2/19 (11%) (P = 0.04) in univariate analysis [144]. Inconclusive (2 studies) [59, 60] Early administration – higher plasma viral load, no difference in severity [59] Possible adverse effect (1 study): osteonecrosis [10]‡. Immunotherapy Convalescent plasma Inconclusive (1 study) CFR [0/1 (0%) vs. 2/28 (7%)] and [0/19 (0%) vs. 5/21 (24%) in 2 comparative studies, and 0/1, 0/1, 0/3, and 10/80 in 4 non-camparative studies [12]‡ 1 study [168] Inconclusive (1 study): 2 out of 3 cases with respiratory failure showed neutralizing activity [169]. Monoclonal antibody 201 [170] m336 [168], hMS-1 [171], 4C2h [172], HR2P-M2 [156]. Other drugs β-D-N4-hydroxycytidine [50]; calpain inhibitor VI [50] Camostat [173] Camostat [80], loperamide [51], chlorpromazine [51, 52, 174], imatinib [174, 175], saracatinib [80, 175], baricitinib [80], dasatinib [174], cyclosporine [176], EST [80], cathepsin L/K inhibitor [80], gemcitabine/toremifene/ triflupromazine [174], mycophenolic acid [155]. β-D-N4-hydroxy- cytidine [50]; calpain inhibitor VI [50] Toremifene [52] (Effective; bold Not effective). In the outcome description, the former is the data of the treatment group and the latter is the data of the control group. CFR: case-fatality ratio; CI: confidence interval; EST: (23,25)-trans-epoxysuccinyl-l-leucylamindo-3-methylbutane ethyl ester; HR: hazard ratio; ICU: intensive care unit; IFN: interferon; L/r: lopinavir/ritonavir; MERS: Middle East respiratory syndrome; OR: odds ratio; RCT: randomized controlled trial; RR: risk ratio; SARS: severe acute d 1213 Theranostics 2021, Vol. 11, Issue 3 Inconclusive (1 study): 2 out of 3 cases with respiratory failure showed neutralizing activity [169]. Umifenovir y y is the only published randomized controlled trial in this table. †A mismatched double-stranded RNA interferon inducer. ‡Meta-analysis. y y study is the only published randomized controlled trial in this table. †A mismatched double-stranded RNA interferon inducer. ‡Meta-anal nistration This study is the only published randomized controlled trial in this table. †A mismatched double-stranded RNA interferon inducer. ‡Meta-analysis. §Intranasal administration. Table 2. Therapeutic agents that showed effects against SARS-CoV-2 in in vitro studies Therapeutics First author Findings Conclusion Antiviral agents Umifenovir Wang [33] EC50 = 4.11 μM; CC50 = 31.79 μM; SI = 7.73 Potent Remdesivir Wang [29] EC50 = 0.77 μM; CC50 > 100 μM; SI > 129.87 Potent Choy [30] EC50 = 26.9 μM; CC50 > 100 μM Not potent Nelfinavir Musarrat [49] Complete inhibition of SARS CoV-2 mediated cell fusion at 10 μM Potent Antiparasitic agents Ivermectin Caly [177] 5000-fold reduction in viral RNA at 48h after a single administration (IC50 < 2mM) Potent Emetine Choy [30] EC50 = 0.5 μM; CC50 = 56.46 μM Potent 4-aminoquinoline (anti-malarial agents) Chloroquine Wang [29] EC50 = 1.13 μM; CC50 > 100 μM; SI > 88.5 Potent Yao [53] Incubation time may influence antiviral activity (24h EC50 = 23.9 μM; 48h EC50 = 5.47 μM). Potent Liu [54] EC50 = 2.71 (MOI = 0.01), 3.81 (0.02), 7.14 (0.2), 7.36 (0.8) μM; CC50 = 273.2 μM Potent Hydroxychloroquine Yao [53] 24h EC50 = 6.14 μM; 48h EC50 = 0.72 μM Potent Liu [54] EC50 = 4.51 (MOI = 0.01), 4.06 (0.02), 17.31 (0.2), 12.96 (0.8) μM; CC50 = 249.5 μM Potent Other agents Homoharrngtonine Choy [30] EC50 = 2.14 μM; CC50 = 59.75 Potent Nitazoxanide Wang [29] EC50 = 2.12 μM; CC50 > 35.53 μM; SI > 16.76 Potent Immunotherapy EK1C4 Xia [178] IC50 = 36.5 nM; CC50 > 5 μM; SI > 136 Potent CC50: 50% cytotoxic concentration; COVID-19: coronavirus disease 2019; EC50: 50% maximal effective concentration; MOI: multiplicity of infection; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; SI: selectivity index. Table 2. Therapeutic agents that showed effects against SARS-CoV-2 in in vitro studies CC50: 50% cytotoxic concentration; COVID-19: coronavirus disease 2019; EC50: 50% maximal effective concentration; MOI: multiplicity of infection; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; SI: selectivity index. http://www.thno.org 1214 Table 3. Summary of RCTs evaluating therapeutics for COVID-19 Table 3. Umifenovir Summary of RCTs evaluating therapeutics for COVID-19 Therapeutics (daily dosage mg) [Common treatment applied to all participants] First author Condition Region Period of enrollment No. of participants (treatment - control group) Outcome of patients or findings: treatment group vs. control group (number of participants or the median value [IQR]) Conclusion Antiviral agents Remdesivir 200 mg (day 1); 100 mg (day 2-10) vs. standard treatment Beigel [20] Not specified World- wide Feb 21- Apr 19 541 - 521 Improvement in the ordinal score on day 15: OR 1.50, 95% CI 1.18 to 1.91 (P = 0.001) 14-day mortality: 32 (6%) vs. 54 (10%) (HR 0.70, 95% CI 0.47 to 1.04) 14-day mortality in patients with a baseline ordinal score of 5 (requiring oxygen): 4/222 (2%) vs. 19/199 (10%) (HR 0.22, 95% CI 0.08 to 0.58) Time to recovery: 11 (95% CI 9–12) vs. 15 (13–19) days (RR for recovery 1.32, 95% CI 1.12 to 1.55; P < 0.0001). Effective A: 200 mg (day 1); 100 mg (day 2-10) B: 200 mg (day 1); 100 mg (day 2–5). C: Standard treatment. Spinner [23] Moderate The US, Europe, Asia Mar 15- Apr 18 197 (A) 199 (B) 200 (C) Better clinical status distribution on the 7-category ordinal scale on day 11: OR (B vs. C) 1.65 (95% CI 1.09 to 2.48) 28-day mortality: 3 (A, 2%); 2 (B, 1%); 4 (C, 2%). Effective (5-day treatment) 200 mg (day 1); 100 (day 2-10) vs. standard treatment Wang [21] Severe China Feb 6- Mar 12 158 - 78 28-day mortality: 22 (14%) vs. 10 (13%) Time to clinical improvement (a 2-point reduction on a 6-category ordinal scale, or discharge from hospital): 21 [13–28] vs. 23 [15–28] days (HR 1.23, 95% CI 0.87 to 1.75) Not effective A: 200 mg (day 1); 100 (day 2-5) B: 200 mg (day1); 100 (day 2-10) Goldman [24] Severe World- wide† Mar 6- Mar 26 200 (A) 197 (B) Clinical improvement of 2 points or more on a 7-category ordinal scale within 14 days: 129 (A, 64%) vs. 107 (B, 54%) (difference -6.5%, 95% CI -15.7% to 2.8%) 14-day mortality among patients receiving MV or ECMO: 10/25 (A, 40%) vs. 7/41 (B, 17%) Serious adverse event: 42 (A, 21%) vs. 68 (B, 35%) (difference 10.8%, 95% CI 2.4% to 19.2%). Favors 5-day treatment Sofosbuvir/daclatasvir 400 mg/60 mg for 14 days vs. Umifenovir standard treatment [hydroxychloroquine with or without lopinavir /ritonavir] Sadeghi [27] Moderate/ severe Iran Mar 26- Apr 26 33 - 33 Duration of hospitalization: 6 [4–8] vs. 8 days [5–13] (P = 0.029) The cumulative incidence of hospital discharge was higher in the treatment group (P = 0.041). Clinical recovery within 14 days: 29 (88%) vs. 22 (67%) (P = 0.076). Effective 400 mg/60 mg plus ribavirin (1,200) vs. hydroxychloroquine and lopinavir/ritonavir with/without ribavirin Abbaspour Kasgari [28] Moderate Iran Mar 20- Apr 8 24 - 24 ICU admission: 0 (0%) vs. 4 (17%) (P = 0.109) Hospital mortality 0 (0%) vs. 3 (13%) (P = 0.234). The cumulative incidence of recovery was higher in the treatment group (P = 0.033). Effective Favipiravir A: 3,200 mg (day 1); 1,200 (day 2–14); B: 3,600 day (day 1); 1,600 (day 2–14); C: Standard treatment. Ivashchenko [31] Moderate Russia Apr–May 20 (A) 20 (B) 20 (C) Discharge or achievement of score 2 on WHO-OSCI by day 15: 13 (A, 65%), 17 (B, 85%), and 17 (C, 85%) Viral clearance on day 5: 25/40 (A and B, 63%) vs. 6/20 (C, 30%) (P = 0.018) Possible effect 3,600 mg (day 1); 1,600 mg (day 2–10) vs. 3,600 mg (day 6); 1,600 mg (day 7–15) Doi [32] Mild Japan Mar 2- May18 36 - 33 Time to discharge from the hospital: 14.0 vs.21.5 days (HR 2.68, 95% CI 1.67 to 4.29) Viral clearance on day 6: 67% vs. 56% (adjusted HR 1.42, 95% CI 0.76 to 2.62) 69 out of 82 participants (84%) developed hyperuricemia. Favors early treatment Other antiviral agents Lopinavir (800)/ ritonavir (200) for 14 days vs. standard treatment Cao [40] Severe China Jan 18-Feb 3 99 - 100 28-day mortality: 19 (19%) vs. 25 (25%) (difference -5.8%; 95% CI -17.3% to 5.7%) Time to clinical improvement (a 2-point reduction on a 7-category ordinal scale or discharge from hospital): 16 [13–17] vs. 16 [15–18] days (HR 1.24, 95% CI 0.90 to 1.72) Hospital stay: 14 [12–17] vs. Umifenovir 16 [13–18] days (difference 1, 95% CI 0 to 2) Not effective A: Lopinavir (400) /ritonavir (100) for 7–14 days B: Umifenovir (600) for 7–14 days C: Standard treatment Li [34] Mild/ moderate China Feb 1–Mar 28 34 (A) 35 (B) 17 (C) Deterioration to severe/critical COVID-19 on day 7: 8/34 (A, 24%), 3/35 (B, 9%), and 2/17 (C, 12%) (P = 0.206) Time to viral clearance: 9.0 (A; SD 5.0), 9.1 (B; 4.4), and 9.3 (C; 5.2) days (P = 0.981) Viral clearance within 7 days: 12/34 (A, 35%), 13/35 (B, 37%), and 7/17 (C, 41% ) (P = 0.966). Not effective A: Ribavirin (2,000 mg l di 1 200 1 800 f Huang [41] Mild/ d t China Jan 29–Feb 25 33 (A) 36 (B) Deterioration to severe COVID-19: 1 (A, 3%), 2 (B, 6%), d 2 (C 6%) (P 0 58) Not effective Sofosbuvir/daclatasvir 400 mg/60 mg for 14 days vs. standard treatment [hydroxychloroquine with or without lopinavir /ritonavir] Sadeghi [27] Moderate/ severe Iran Mar 26- Apr 26 33 - 33 400 mg/60 mg plus ribavirin (1,200) vs. hydroxychloroquine and lopinavir/ritonavir with/without ribavirin Abbaspour Kasgari [28] Moderate Iran Mar 20- Apr 8 24 - 24 Favipiravir A: 3,200 mg (day 1); 1,200 (day 2–14); B: 3,600 day (day 1); 1,600 (day 2–14); C: Standard treatment. Ivashchenko [31] Moderate Russia Apr–May 20 (A) 20 (B) 20 (C) 3,600 mg (day 1); 1,600 mg (day 2–10) vs. 3,600 mg (day 6); 1,600 mg (day 7–15) Doi [32] Mild Japan Mar 2- May18 36 - 33 Duration of hospitalization: 6 [4–8] vs. 8 days [5–13] (P = 0.029) The cumulative incidence of hospital discharge was higher in the treatment group (P = 0.041). Clinical recovery within 14 days: 29 (88%) vs. 22 (67%) (P = 0.076). Effective ICU admission: 0 (0%) vs. 4 (17%) (P = 0.109) Hospital mortality 0 (0%) vs. 3 (13%) (P = 0.234). The cumulative incidence of recovery was higher in the treatment group (P = 0.033). Effective 28-day mortality: 19 (19%) vs. 25 (25%) (difference -5.8%; 95% CI -17.3% to 5.7%) Time to clinical improvement (a 2-point reduction on a 7-category ordinal scale or discharge from hospital): 16 [13–17] vs. 16 [15–18] days (HR 1.24, 95% CI 0.90 to 1.72) Hospital stay: 14 [12–17] vs. Umifenovir 16 [13–18] days (difference 1, 95% CI 0 to 2) Not effective Deterioration to severe/critical COVID-19 on day 7: 8/34 (A, 24%), 3/35 (B, 9%), and 2/17 (C, 12%) (P = 0.206) Time to viral clearance: 9.0 (A; SD 5.0), 9.1 (B; 4.4), and 9.3 (C; 5.2) days (P = 0.981) Viral clearance within 7 days: 12/34 (A, 35%), 13/35 (B, 37%), and 7/17 (C, 41% ) (P = 0.966). Not effective Deterioration to severe COVID-19: 1 (A, 3%), 2 (B, 6%), and 2 (C, 6%) (P = 0.58) Time to viral clearance: 13.0 [9.0–25.5] (A), 12.0 [7.0– 19.0] (B), and 15.0 [9.3–17.8] (C) days (P = 0.42) Viral clearance on day 14: 17/33 (A, 52%), 22/36 (B, 61%), and 15/32 (C, 47%). Not effective A: Lopinavir (400) /ritonavir (100) for 7–14 days B: Umifenovir (600) for 7–14 days C: Standard treatment Li [34] Mild/ moderate China Feb 1–Mar 28 34 (A) 35 (B) 17 (C) A: Ribavirin (2,000 mg loading; 1,200–1,800 mg for 14 days) B: Lopinavir (800)/ritonavir (200) C: Ribavirin plus lopinavir/ritonavir Huang [41] Mild/ moderate China Jan 29–Feb 25 33 (A) 36 (B) 32 (C) Azvudine (FNC) (5) vs. standard treatment Ren [45] Mild/ moderate China Feb 18–Feb 29 10 - 10 http://www.thno.org 1215 Theranostics 2021, Vol. 11, Issue 3 Number of hospitalized participants: 8 (6%) vs. 11 (7%) (RR 0.75, 95% CI 0.32 to 1.77); Time to the resolution of symptoms : 10 [4–18] vs. 12 [6-21] days (P = 0.38); Reduction in viral load on day 7: -3.49 (SD 0.20) vs. -3.37 (0.19) log10 copies/mL (difference -0.12, 95% CI -0.25 to 0.5). Not effective Alleviation of symptoms by day 28¶: 60% vs. 67% (difference -7%, 95% CI -41% to 28%) Viral clearance by day 28: 56 (75%) vs. 53 (71%) Adverse events: 21/70 (30%) vs. 7/80 (9%). Not effective Initiation of MV: 4 (4%) vs. 5 (5%) (P = 0.75) 28-day mortality: 6 (6%) vs. 5 (5%) (P = 0.77). Not effective Change in symptom severity score over 14 days: -2.21(SE 0.23) vs. -2.10 (0.23) (P = 0.51). Not effective 1,200 mg (day 1–3); 800 mg (day 4–14) vs. standard treatment Tang [180] Mild/ moderate China Feb 11–Feb 29 75 - 75 800 mg (day 1); 400mg (day 2–15) vs. standard treatment Abd-Elsalam b [181] Not specified Egypt Mar–Jun 97 - 97 1,400 mg (day 1); 600 mg (day 2–5) vs. Umifenovir standard treatment Skipper [182] Mild The US, Canada Mar 22– May 20 73 - 72 800 mg (day 1); 400 mg (day 2–5) vs. standard treatment Kamran [56]†† Mild Pakistan Apr 10– May 31 349 - 151 400 mg for 5 days vs. standard treatment Chen [55]†† Mild/ moderate China Feb 4- Feb 28 31 - 31 800 mg (day 1); 400 mg (day 2–7) vs. standard treatment Chen [183]†† Mild/ moderate Taiwan Apr 1- May 31 21 - 12 Disease progression‡‡: 11 (3%) vs. 5 (3%) (P = 0.865) Viral clearance within 7 days: 182 (52%) vs. 54 (36%) (P = 0.001). Possible effect Improvement of chest CT scans on day 6: 25 (81%) vs. 17 (55%) (P = 0.0476) Duration of fever: 2.2 (SD 0.4) vs. 3.2 (1.3) days (P = 0.0008). Effective Clinical recovery (3 consecutive negative results of viral PCR and resolution of major symptoms) within 14 days: 6/21 (29%) vs. 5/12 (42%) (P = 0.51) Time to viral clearance: 5 (95% CI 1 to 9) vs. 10 (2 to 12) days (P = 0.40) Viral clearance within 14 days: 17/21 (81%) vs. 9/12 (75%) (P = 0.36). Not effective Disease progression‡‡: 11 (3%) vs. 5 (3%) (P = 0.865) Viral clearance within 7 days: 182 (52%) vs. 54 (36%) (P = 0.001). Possible effect Improvement of chest CT scans on day 6: 25 (81%) vs. 17 (55%) (P = 0.0476) Duration of fever: 2.2 (SD 0.4) vs. 3.2 (1.3) days (P = 0.0008). Effective Clinical recovery (3 consecutive negative results of viral PCR and resolution of major symptoms) within 14 days: 6/21 (29%) vs. 5/12 (42%) (P = 0.51) Time to viral clearance: 5 (95% CI 1 to 9) vs. 10 (2 to 12) days (P = 0.40) Viral clearance within 14 days: 17/21 (81%) vs. 9/12 (75%) (P = 0.36). Not effective Azithromycin 500 mg for 10 days vs. standard treatment Furtado [58] Severe Brazil Mar 28- May 19 214 - 183 500 mg for 5 days vs. standard treatment [Lopinavir/ritonavir and hydroxychloroquine] Sekhavati [57] Not specified Iran Apr 24- May 8 56 - 55 Colchicine 2 mg (day 1)§§; 1 mg (till discharge or day 21) vs. standard treatment [Chloroquine or hydroxychloroquine and azithromycin]¶¶ Deftereos [70] Not specified Greece Apr 3- Apr 27 55 - 50 1.5 mg (day 1–5); 1 mg (day 6–10) vs. standard treatment [azithromycin, hydroxychloroquine, and unfractionated heparin]. Theranostics 2021, Vol. 11, Issue 3 Theranostics 2021, Vol. 11, Issue 3 weekly) vs. standard treatment [hydroxychloroquine, azithromycin] CM4620-IE (Auxora: calcium release-activated calcium channel inhibitor) (2.0 mg/kg/day continuous infusion on day 1; 1.6 mg/kg/day on day 2–3) vs. standard treatment. Miller [79] Severe/ critical The US Apr 8- May 13 20 – 10 IMV or death by day 30: 3/17 (18%) vs. 5/9 (56%) in participants with severe COVID-19 (HR 0.23, 95% CI 0.05 to 0.96; P < 0.05) The mean difference in 8-point ordinal scale between groups was statistically significant at day 6 and day 9– 12 (P < 0.05). Effective Ruxolitinib (Janus- associated kinase inhibitors) (10) vs. standard treatment Cao [81] Severe China Feb 9- Feb 28 20 - 21 Improvement of chest CT scans on day 14: 18 (90%) vs. 13 (62%) (P = 0.0495) 28-day mortality: 0 (0%) vs. 3 (14%) (P = 0.232) Time to clinical improvement (a 2-point reduction on a 7-category ordinal scale or discharge from hospital): 12 [10–19] vs. 15 [10–18] days (P = 0.147) (HR 1.669, 95% CI 0.836 to 3.335) Time to lymphocyte recovery: 5 [2–7] vs. 8 [2–11] days (P = 0.033). Effective Leflunomide (DHODH inhibitor) (100 day 1–3; 20 day 4–10) vs. standard treatment [Umifenovir]. Hu [82] Moderate China Feb 20- Feb 28 5 -5 Duration of viral shedding: 5 vs. 11 days (P = 0.046) The difference in the level of serum CRP measured before treatment and on day: 32 [5.6–not tested] vs.0 [0-9.1] mg/L (P = 0.047). Possible effect Immunotherapy Interferon IFN-β1a (12 million IU 3 times weekly for 2 weeks) vs. standard treatment [Hydroxychloroquine plus lopinavir/ritonavir or atazanavir/ritonavir]. Davoudi- Monfared [83] Severe Iran Feb 29- Apr 3 42 - 39 28-day mortality: 19% vs. 44% (P = 0.015) Rate of discharge from the hospital: 67% vs. 44% (OR 2.5, 95% CI 1.05 to 6.37) Early administration of IFN-β1a reduced mortality (OR 13.5, 95% CI 1.5 to 118). Time to clinical improvement: 9.7 ± 5.8 vs. 8.3± 4.9 days (P = 0.95). Effective IFN-β1b (3 doses of 8 million IU on alternate days) plus ribavirin (800) for 14 days vs. standard treatment [Lopinavir/ritonavir] Hung [44] Mild/ moderate Hong Kong Feb 10- Mar 20 86 - 41 Time to a NEWS2 of 0: 4 [3–8] vs. 8 [7–9] days (HR 3.92, 95% CI 1.66 to 9.23) Time to a SOFA score of 0: 3.0 [1.0–8.0] vs. Theranostics 2021, Vol. 11, Issue 3 8.0 [6.5–9.0] days (HR 1.89, 95% CI 1.03 to 3.49) Length of hospital stay: 9.0 [7.0–13.0] vs. 14.5 [9.3–16.0] days (HR 2.72, 95% CI 1.2 to 6.13) Time to viral clearance: 7 [5–11] vs. 12 [8–15] days (HR 4.37, 95% CI 1.86 to 10.24, P = 0.001). Effective IFN-β1b (250 mcg on alternate days for 2 weeks) vs. standard treatment Rahmani [84] Severe Iran Apr 20- May 20 33- 33 Discharge from hospital by day 14: 26 (79%) vs. 18 (55%) (OR 3.09, 95% CI 1.05 to 9.11, P = 0.03) ICU admission: 14 (42%) vs. 22 (67%) (P = 0.04) Time to clinical improvement (a 2-point reduction on a 6-category ordinal scale): 9 [6–10] vs. 11 [9–15] days (P = 0.002). Effective Inhaled IFN-κ (2) plus TFF2 (5) for 6 days vs. standard treatment Fu [85] Moderate China Mar 23- May 23 40 - 40 Time to improvement of chest CT: 6.2 (95% CI 5.1–7.3) vs. 8.8 (95% CI 7.6-10.0) days (P = 0.002) Time to viral clearance: 3.8 (95% CI 2.1–5.5) vs. 7.4 (95% CI 4.6–10.2) days (P = 0.031). Effective A: Novaferon (40 mcg) B: Novaferon and lopinavir (800)/ritonavir (200) C: Lopinavir/ritonavir. Zheng [86] Moderate/ severe China Feb 1- Feb 20 30 (A) 30 (B) 29 (C) Viral clearance on day 6: 15/30 (A, 50%; P = 0.04) or 18/30 (B, 60%; P = 0.0053) vs. 7/29 (C, 24%) Time to viral clearance: 6 (A, P = 0.417) or 6 (B, P = 0.036) vs. 9 (C) days. Possible effect Convalescent plasma 4-13 mL/kg vs. standard treatment Li [87] Severe/ life-threateni ng China Feb 14– Apr 1 52 - 51 Clinical improvement (a 2-point reduction on a 6-category ordinal scale or discharge from hospital) within 28 days: 27 (52%) vs. 22 (43%) (HR 1.40, 95% CI 0.79 to 2.49) Clinical improvement within 28 days for the participants with severe COVID-19: 21/23 (91%) vs. 15/22 (28%) (HR 2.15, 95% CI 1.07 to 4.32) 28-day mortality: 8 (16%) vs. 12 (24%) (OR 0.59, 0.22 to 1.59) Viral clearance within 72 hours: 41 (87%) vs. 15 (38%) (OR 11.39, 95% CI 3.91 to 33.18). Effective (severe COVID-19 subgroup) 200 mL (day1–2) vs. standard treatment Agarwal [89]†† Moderate India Apr 22- Jul 14 235 - 229 28-day mortality: 34 (15%) vs. 31 (14%) (adjusted OR 1.06, 95% CI 0.61 to 1.83) Disease progression (PaO2/FiO2 < 100): 44 (19%) vs. Umifenovir Lopes [71]†† Moderate/ severe Brazil Apr 11- Jul 06 17 - 18 Other agents Methylprednisolone (250 for 3 days) vs.standard treatment [Hydroxychloroquine, lopinavir, naproxen]. Edalatifard [66] Severe Iran Apr 20– Jun 20 34 - 28 Telmisartan (160) for 14 days vs. standard treatment Duarte [72]†† Not specified Argenti na May 14– Jul 30 41 - 41 Enoxaparin (0.75–2 mg/kg for 4–14 days) vs. enoxaparin (40 or 80) or unfractionated heparin (15,000–22,500 IU)††† Lemos [75] Severe and intubated Brazil Apr–Jul 10 - 10 Calcifediol (0.532 on day 1; 0.266 on day 3 and 7, then Entrenas Castillo [78] Not specified Spain Not specified 50 - 26 Azithromycin 500 mg for 10 days vs. standard treatment Furtado [58] Severe Brazil Mar 28- May 19 214 - 183 500 mg for 5 days vs. standard treatment [Lopinavir/ritonavir and hydroxychloroquine] Sekhavati [57] Not specified Iran Apr 24- May 8 56 - 55 Colchicine 2 mg (day 1)§§; 1 mg (till discharge or day 21) vs. standard treatment [Chloroquine or hydroxychloroquine and azithromycin]¶¶ Deftereos [70] Not specified Greece Apr 3- Apr 27 55 - 50 1.5 mg (day 1–5); 1 mg (day 6–10) vs. standard treatment [azithromycin, hydroxychloroquine, and unfractionated heparin]. Lopes [71]†† Moderate/ severe Brazil Apr 11- Jul 06 17 - 18 Worse clinical status on the 6-category ordinal scale on day 15: OR 1.36 (95% CI 0.94 to 1.97, P = 0.11) 28-day mortality: 90 (42%) vs. 73 (40%) (HR 1.08, 95% CI 0.79 to 1.47, P = 0.63). Not effective Length of hospital stay: 4.6 (SD 2.6) vs. 6.0 (SD 3.2) days (P = 0.02) Mortality: 0 (0%) vs. 1 (2%) (P = 0.495) ICU admission: 2 (4%) vs. 7 (13%) (P = 0.070). Effective http://www.thno.org 1216 Theranostics 2021, Vol. 11, Issue 3 Defined as a Borg score > 3, improved dyspnea, stopped fever for 72 hours, SO2 > 93%, tolerated oral regimen, normal urinary output, and reduced C-reactive protein level without any side effects. †††The dosage was determined according to age, body weight, and creatinine clearance. ‡‡‡The deferred treatment group received convalescent plasma only when a PaO2/FiO2 < 200 criterion was met during hospitalization or when the patient still required hospitalization for symptomatic COVID-19 more than 7 days after enrollment. §§§Progression to acute respiratory distress syndrome, sepsis, or septic shock. Table 4. Summary of meta-analyses evaluating therapeutics for COVID-19 http://www thno org Table 4. Summary of meta-analyses evaluating therapeutics for COVID-19 Comparisons First author No. of studies No. of participants Type of metrics Model Summary effect (95% CI) P I2 (P) Publication bias Conclusion Antiviral agents Remdesivir Mortality Misra [22] 2 [20, 21] 0 54 (54)/ 696 (696) 64 (64)/ 599 (599) RR Random 0.74 (0.40 to 1.37) NA 58% (0.12) NA Not effective Clinical recovery Misra [22] 2 [20, 21] 0 437 (437)/ 696 (696) 318 (318)/ 599 (599) RR Fixed 1.17 (1.07 to 1.29) NA 0% (0.70) NA Effective Clinical improvement (5 vs. Theranostics 2021, Vol. 11, Issue 3 Vilobelimab (anti-C5a antibody IFX-1) 800 mg (day 1, 2, 4, 8, and 15) vs. standard treatment Vlaar [92] Severe Nether-l ands Mar 31- Apr24 15 - 15 CIGB-325 (anti-CK2) 2.5 mg/kg (day 1–5) vs. standard treatment Cruz [93]†† Not specified Cuba Jun 1- Jun 16 10 - 10 CI: confidence interval; COVID-19: coronavirus disease 2019; CRP: C-reactive protein; CT: computed tomography; DHODH: dihydroorotate dehydrogenase; HR: hazard ratio; ICU: intensive care unit; IFN: interferon; IMV: invasive mechanical ventilation; IQR: interquartile range; IU: international unit; MV: mechanical ventilation; NEWS2: National Early Warning Score 2; OSCI: ordinal scale for clinical improvement; OR: odds ratio; PCR: polymerase chain reaction; RCT: randomized controlled trial; rhG-CSF: Recombinant human granulocyte colony-stimulating factor; RR: relative risk; SD: standard deviation; SE: standard error; SOFA: sequential organ failure assessment; WHO: World Health Organization. All of the presented studies were conducted in 2020. In the outcome description, the former is the data of the treatment group and the latter is the data of the control group. The United States, Denmark, the United Kingdom, Greece, Germany, South Korea, Mexico, Spain, Japan, and Singapore. †The United States, Italy, Spain, Germany, Hong Kong, Singapore, South Korea, and Taiwan. ‡750 mg for participants with a mild or ordinary condition or 1,000 mg for participants with a severe or critical condition. §Defined as normalization of body temperature, respiratory rate, oxygen saturation, cough, and absorption of pulmonary infection on chest CT. ¶Resolving from fever to an axillary temperature of 36.6°C or below, normalization of SpO2 (> 94% on room air), and disappearance of respiratory symptoms including nasal congestion, cough, sore throat, sputum production, and shortness of breath. The most common adverse event in the treatment group was diarrhea (7/70). ††Preprints from Medrxiv.org. ‡‡Defined as development of fever higher than 101 F for more than 72 hours, shortness of breath by minimal exertion (10-Step walk test), derangement of basic laboratory parameters (absolute lymphocyte count < 1,000 mm3 or raised serum C-reactive protein level), or appearance of infiltrates on chest radiograph during course of treatment. §§In the case of azithromycin coadministration, a single 1.0-mg loading dose of colchicine was administered. ¶¶Chloroquine or hydroxychloroquine was administered to 100% and 96% of participants in the treatment and the control group, respectively. Azithromycin was administered to 93% and 92% of participants in the treatment and the control group, respectively. Theranostics 2021, Vol. 11, Issue 3 41 (18%) (adjusted OR 1.09, 95% CI 0.67 to 1.77). Not effective 300 mL vs. standard treatment Gharbharan [184]†† Not specified Nether-l ands Apr 8- Jun 10 43 - 43 60-day mortality: 6 (14%) vs. 11 (26%) (OR 0.95, 95% CI 0.20 to 4.67) Improvement in WHO-OSCI on day 15: 25 (58%) vs. 25 (58%) (OR 1.30, 95% CI 0.52 to 3.32). Not effective 250–300 mL vs. standard Avendaño-So Not Spain Apr 4- 38 - 43 Initiation of MV or death by day 15: 0 (0%) vs. 6 (14%) Effective weekly) vs. standard treatment [hydroxychloroquine, azithromycin] CM4620-IE (Auxora: calcium release-activated calcium channel inhibitor) (2.0 mg/kg/day continuous infusion on day 1; 1.6 mg/kg/day on day 2–3) vs. standard treatment. Miller [79] Severe/ critical The US Apr 8- May 13 20 – 10 Ruxolitinib (Janus- associated kinase inhibitors) (10) vs. standard treatment Cao [81] Severe China Feb 9- Feb 28 20 - 21 Inhaled IFN-κ (2) plus TFF2 (5) for 6 days vs. standard treatment Fu [85] Moderate China Mar 23- May 23 40 - 40 A: Novaferon (40 mcg) B: Novaferon and lopinavir (800)/ritonavir (200) C: Lopinavir/ritonavir. Zheng [86] Moderate/ severe China Feb 1- Feb 20 30 (A) 30 (B) 29 (C) Convalescent plasma 4-13 mL/kg vs. standard treatment Li [87] Severe/ life-threateni ng China Feb 14– Apr 1 52 - 51 200 mL (day1–2) vs. standard treatment Agarwal [89]†† Moderate India Apr 22- Jul 14 235 - 229 300 mL vs. standard treatment Gharbharan [184]†† Not specified Nether-l ands Apr 8- Jun 10 43 - 43 250–300 mL vs. standard treatment Avendaño-So là [88]†† Not specified Spain Apr 4- Jul 10 38 - 43 200 mL (day1–2) vs. deferred treatment‡‡‡ Barcells [185]†† At risk for progression Chile May 10- Jul 18 28 - 30 http://www.thno.org 1217 Theranostics 2021, Vol. 11, Issue 3 Other immunotherapies rhG-CSF 5 mcg/kg (day 1–3) vs. standard treatment Cheng [90] Lympho- penia China Feb 18- Apr 10 100 - 100 21-day mortality: 2 (2%) vs. 10 (10%) (HR 0.19, 95% CI 0.04 to 0.88) Disease progression§§§: 2 (2%) vs. 15 (15%) (difference -13%, 95% CI -21.4% to -5.4%) Time to clinical improvement (a 1-point reduction on a 7-category ordinal scale or discharge from hospital): 12 [10–16] vs. 13 [11–17] (HR 1.28, 95% CI 0. 95-1.71, P = 0.06). Effective (lympho- penia) Intravenous immunoglobulin 0.5g/kg/day for 3 days plus methylprednisolone (40 mg once) vs. Theranostics 2021, Vol. 11, Issue 3 standard treatment Sakoulas [91]†† Moderate/ severe (except patients with MV) The US May 1- Jun 16 16 - 17 (Among subjects with alveolar-arterial oxygen gradient of >200 mmHg at enrollment) Initiation of MV within 30 days: 2/14 (14%) vs. 7/12 (58%) (P = 0.038), Length of hospital stay: 11 (range 5–22) vs. 19 (4–30) days (P = 0.013) Length of ICU stay: 2.5 (range 0–16) vs. 12.5 (1–29) days (P = 0.006) Difference in PaO2/FiO2 on day 7: +131 (+35 to +330) vs.+44.5 (-115 to +157) (P = 0.01). Effective Vilobelimab (anti-C5a antibody IFX-1) 800 mg (day 1, 2, 4, 8, and 15) vs. standard treatment Vlaar [92] Severe Nether-l ands Mar 31- Apr24 15 - 15 28-day mortality: 2 (13%) vs. 4 (27%) (adjusted HR 0.65, 95% CI 0.10 to 4.14) Difference in the change in PaO2/FiO2on day 5 (least squares mean): 17% (SD 63) vs. 41% (difference -24%, 95% CI -58% to 9%, P = 0.15). Not effective CIGB-325 (anti-CK2) 2.5 mg/kg (day 1–5) vs. standard treatment Cruz [93]†† Not specified Cuba Jun 1- Jun 16 10 - 10 Reduction in the number of pulmonary lesions on the chest CT: 5/6 (83%) vs. 3/7 (43%) (Bayesian P (difference > 0) = 0.951). Time to viral clearance: 11 (SD 8) vs. 12 (SD 6) days (P = 0.614). Effective Other immunotherapies rhG-CSF 5 mcg/kg (day 1–3) vs. standard treatment Cheng [90] Lympho- penia China Feb 18- Apr 10 100 - 100 Other immunotherapies rhG-CSF 5 mcg/kg (day 1–3) vs. standard treatment Cheng [90] p rhG-CSF 5 mcg/kg (day 1–3) vs. standard treatment Cheng [90] Intravenous immunoglobulin 0.5g/kg/day for 3 days plus methylprednisolone (40 mg once) vs. standard treatment Sakoulas [91]†† Moderate/ severe (except patients with MV) The US May 1- Jun 16 16 - 17 vs.+44.5 (-115 to +157) (P = Vilobelimab (anti-C5a antibody IFX-1) 800 mg (day 1, 2, 4, 8, and 15) vs. standard treatment Vlaar [92] Severe Nether-l ands Mar 31- Apr24 15 - 15 28-day mortality: 2 (13%) v 95% CI 0.10 to 4.14) Difference in the change in squares mean): 17% (SD 63 95% CI -58% to 9%, P = 0.15 CIGB-325 (anti-CK2) 2.5 mg/kg (day 1–5) vs. standard treatment Cruz [93]†† Not specified Cuba Jun 1- Jun 16 10 - 10 Reduction in the number o chest CT: 5/6 (83%) vs. 3/7 (difference > 0) = 0.951). Time to viral clearance: 11 0.614). Theranostics 2021, Vol. 11, Issue 3 10-days of treatment) Jiang [26] 2 [24, 25] 0 263 (263)/ 391 (391) 233 (233)/ 390 (390) OR Random 1.33 (1.01 to 1.76) NA NA NA Favors 5-day treatment Adverse events Misra [22] 2 [20, 21] 0 258 (258)/ 696 (696) 222 (222)/ 599 (599) RR Fixed 0.91 (0.79 to 1.05) NA 7% (0.30) NA Inconclusive Serious adverse events Juul [186] 2 [20, 21] 0 142 (142)/ 554 (554) 161 (161)/ 438 (438) RR Random 0.77 (0.63 to 0.94) 0.01 0.0% (0.66) NA Inconclusive Favipiravir Clinical improvement by day 14 Shrestha [187] 2 [31, 188] 1 73 (41)/ 84 (49) 49 (21)/75 (30) RR Fixed 1.29 (1.08 to 1.54) 0.005 16% (0.30) NA Effective Shrestha [187] 2 [31, 188] 0 41 (41)/ 49 (49) 21 (21)/30 (30) RR Fixed 1.12 (0.87 to 1.44) 0.37 0% (0.98) NA Not effective Viral clearance by day 14 Shrestha [187] 2 [31, 188] 1 77 (44)/ 84 (49) 61 (28)/75 (30) RR Random 1.06 (0.84 to 1.33) 0.65 67% (0.05) NA Inconclusive Shrestha [187] 2 [31, 188] 0 44 (44)/ 49 (49) 28 (28)/30 (30) RR Random 0.95 (0.74 to 1.22) 0.67 41% (0.19) NA Inconclusive Umifenovir Clinical recovery Misra [22] 1 [34] 1 51 (32)/ 69 (35) 40 (13)/65 (17) RR Fixed 1.08 (0.85 to 1.38) NA 0% (0.42) N Not effective http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 1218 Comparisons First author No. of studies No. Theranostics 2021, Vol. 11, Issue 3 11, Issue 3 Theranostics 2021, Vol. 11, Issue 3 1219 Comparisons First author No. of studies No. Theranostics 2021, Vol. 11, Issue 3 of participants Type of metrics Model Summary effect (95% CI) P I2 (P) Publication bias Conclusion Mortality Das [193] 0 4 NA (0)/ 1145 (0) NA (0)/1165 (0) OR Random 2.84 (2.19 to 3.69) <0.00 001 0% (0.43) Y Harmful Yang [191] 0 3 214 (0)/ 854 (0) 46 (0)/395 (0) OR Fixed 2.34 (1.63 to 3.36) <0.00 001 0% (0.85) N Harmful Deterioration† Yang [191] 0 3 101 (0)/ 840 (0) 25 (0)/414 (0) OR Random 4.97 (0.01 to 4781.7) 0.65 95% (<0.00001) N Not effective Wang [42] 0 2 115 (0)/ 328 (0) 833 (0)/3969 (0) RR Random 0.93 (0.17 to 5.09) NA 94.2% (<0.001) N Not effective Corticosteroids Mortality Lu [67] 0 4 94 (0)/329 (0) 58 (0)/408 (0) RR Random 2.00 (0.69 to 5.75) NA 90% (<0.001) NA Not effective Mortality (severe COVID-19 subgroup) Ye [68] 0 2 NA (0)/ 227 (0) NA (0)/104 (0) HR Random 2.30 (1.00 to 5.29) NA 0% (0.768) NA Not effective Time to viral clearance Sarkar [69] 0 2 82 69 MD Random 1.42 (-0.52 to 3.37) 0.15 0% (0.52) NA Not effective Renin-angiotensin-aldosterone system inhibitors for patients with hypertension Mortality (ACEI) Pranata [73] 0 3 29 (0)/110 (0) 87 (0)/326 (0) OR Random 0.68 (0.39 to 1.17) 0.16 0% (0.62) Y Not effective Mortality (ARB) Pranata [73] 0 3 29 (0)/158 (0) 87 (0)/326 (0) OR Random 0.51 (0.29 to 0.90) 0.02 22% (0.28) Y Effective Mortality (ACEI or ARB) Flacco [74] 0 4 NA (0)/ 921 (0) NA (0)/1491 (0) OR Random 0.88 (0.68 to 1.14) 0.33 24% (0.27) N Not effective Anticoagulants Mortality Lu [76] 0 5 536 (0 )/ 2886 (0) 947 (0)/5647 (0) RR Random 0.86 (0.69 to 1.09) 0.218 47.4% (0.107) NA Not effective Heparin - mortality (severe COVID- 19 subgroup) Abdel-Maboud [77] 0 2 50 (0)/126 (0) 115 (0)/368 (0) RR Random 1.09 (0.84 to 1.42) NA 0% (0.537) NA Not effective Convalescent plasma Mortality Talaie [198] 1 [87] 2 10 (8)/82 (52) 21 (12)/81 (51) RR Random (0.26 to 1.03) NA 0% (0.484) N Not effective Clinical improvement Talaie [198] 1 [87] 2 46 (27)/ 82 (52) 32 (22)/81 (51) RR Random 1.41 (1.01 to 1.98) NA 66.6% (0.050) Y Effective Viral clearance Sarkar [202] 1 [87] 2 54 (41)/ 68 (52) 18 (15)/76 (51) OR Random 11.29 (4.92 to 25.92) <0.00 001 0% (0.40) Y Possible effect Tocilizumab Mortality Lan [203]§ 0 7 39 (0)/241 (0) 85 (0)/352 (0) RR Random 0.61 (0.31 to 1.22) 0.16 68% (0.005) NA Not effective Mortality (lopinavir/riton avir subgroup)¶ Malgie [94] 0 2 7 (0)/94 (0) 22 (0)/56 (0) RD Random -0.31 (-0.57 to -0.05) NA NA Y Effective ICU admission and initiation of MV Lan [203]§ 0 5 47 (0)/134 (0) 44 (0)/279 (0) RR Random 1.51 (0.33 to 6.78) 0.59 86% (<0.00001) NA Not effective ACEI: angiotensin-converting enzyme inhibitor; ARB: angiotensin receptor blocker; ARDS: acute respiratory distress syndrome; CI: confidence interval; CNS: central nervous system; COVID-19: coronavirus disease 2019; ECG: electrocardiogram; HR: hazard ratio; ICU: intensive care unit; MD: mean difference; MV: mechanical ventilation; NA: not applicable; OR: odds ratio; RCT: randomized controlled trial; RD: risk difference; RR: relative risk. Theranostics 2021, Vol. 11, Issue 3 The RCTs included in the meta-analyses of this table were also included in our target RCTs and are presented in Table 3, except an RCT [25] with no peer-reviewed or preprint report released, an RCT [192] published in Chinese, and an RCT [188] in which a statistical analysis was not conducted. A 2-point reduction on a 7-category ordinal scale. †Progression to severe COVID-19. ‡An increase in severity compared to the baseline severity. §One participant in the treatment arm of one included study was diagnosed as suspected COVID-19 with a negative PCR result. ¶All participants received lopinavir plus ritonavir but did not receive corticosteroids. “l i i / it i l IFN ith ith t ACEI: angiotensin-converting enzyme inhibitor; ARB: angiotensin receptor blocker; ARDS: acute respiratory distress syndrome; CI: confidence interval; CNS: central nervous system; COVID-19: coronavirus disease 2019; ECG: electrocardiogram; HR: hazard ratio; ICU: intensive care unit; MD: mean difference; MV: mechanical ventilation; NA: not applicable; OR: odds ratio; RCT: randomized controlled trial; RD: risk difference; RR: relative risk. The RCTs included in the meta-analyses of this table were also included in our target RCTs and are presented in Table 3, except an RCT [25] with no peer-reviewed or preprint report released, an RCT [192] published in Chinese, and an RCT [188] in which a statistical analysis was not conducted. A 2-point reduction on a 7-category ordinal scale. †Progression to severe COVID-19. ‡An increase in severity compared to the baseline severity. §One participant in the treatment arm of one included study was diagnosed as suspected COVID-19 with a negative PCR result. ¶All participants received lopinavir plus ritonavir but did not receive corticosteroids. “lopinavir/ritonavir plus IFN-α with or without ribavirin” with “ribavirin plus IFN-α” [41], treatment with lopinavir/ritonavir did not show superior outcomes in terms of clinical deterioration or viral clearance (Table 3). In meta-analyses on COVID-19 involving two of these RCTs [34, 40], treatment with lopinavir/ritonavir was not associated with clinical recovery or viral clearance [22, 42, 43] (Table 4). Lopinavir/ritonavir In two human non-RCTs on SARS, the treatment group performed better with respect to the overall mortality rate or the incidence of ARDS [36, 37]. An ongoing RCT on MERS involved a combination of lopinavir/ritonavir and IFN-β [19], which has been shown to be effective in two in vivo studies on MERS [38, 39] (Table 1). In an RCT on COVID-19, treatment with lopinavir/ritonavir was not associated with a mortality rate reduction at day 28 (treatment group 19.2% vs. control group 25.0%; difference, -5.8%; 95% CI -17.3% to 5.7%) [40]. In the aforementioned RCT on COVID-19 comparing a combination of lopinavir/ ritonavir and umifenovir with standard treatment [34], and in another RCT on COVID-19 comparing ACEI: angiotensin-converting enzyme inhibitor; ARB: angiotensin receptor blocker; ARDS: acute respiratory distress syndrome; CI: confidence interval; CNS: central nervous system; COVID-19: coronavirus disease 2019; ECG: electrocardiogram; HR: hazard ratio; ICU: intensive care unit; MD: mean difference; MV: mechanical ventilation; NA: not applicable; OR: odds ratio; RCT: randomized controlled trial; RD: risk difference; RR: relative risk. The RCTs included in the meta-analyses of this table were also included in our target RCTs and are presented in Table 3, except an RCT [25] with no peer-reviewed or preprint report released, an RCT [192] published in Chinese, and an RCT [188] in which a statistical analysis was not conducted. A 2-point reduction on a 7-category ordinal scale. †Progression to severe COVID-19. ‡An increase in severity compared to the baseline severity. §One participant in the treatment arm of one included study was diagnosed as suspected COVID-19 with a negative PCR result. ¶All participants received lopinavir plus ritonavir but did not receive corticosteroids. Theranostics 2021, Vol. 11, Issue 3 of participants Type of metrics Model Summary effect (95% CI) P I2 (P) Publication bias Conclusion Viral clearance (on day 14) Huang [35] 1 [34] 4 122 (32)/ 140 (35) 174 (13)/ 247 (17) RR Random 1.27 (1.04 to 1.55) 0.02 63% (0.03) NA Possible effect Adverse events Misra [22] 1 [34] 1 8 (5)/69 (35) 4 (0)/65 (17) RR Fixed 1.80 (0.52 to 6.19) NA 10% (0.29) N Inconclusive Lopinavir/ritonavir Clinical recovery Misra [22] 2 [34, 40] 1 135 (107)/ 185 (133) 110 (83)/ 165 (117) RR Fixed 1.08 (0.94 to 1.24) NA 0% (0.70) N Not effective Viral clearance Wang [42] 2 [34, 40] 1 96 (61)/153 (93) 141 (53)/209 (88) RR Fixed 0.90 (0.76 to 1.07) 0.225 33.9% (0.220) N Inconclusive Liu [43] 2 [34, 40] 0 48 (48)/ 80 (80) 45 (45)/78 (78) RR Random 0.99 (0.76 to 1.29) 0.93 0% (0.74) NA Inconclusive Adverse events Misra [22] 2 [34, 40] 1 67 (58)/ 185 (133) 53 (49)/ 165 (117) RR Random 1.73 (0.57 to 5.26) NA 67% (0.05) N Inconclusive Increased serum creatinine Zhong [189] 1 [40] 1 4 (2)/147 (95) 7 (7)/147 (99) RR Random 0.86 (0.66 to 11.97) NA 61% (0.110) NA Inconclusive Hydroxychloroquine 28-day mortality Elsawah [190] 2 [179, 180] 0 0 (0)/ 239 (239) 0 (0) 264 (264) RD Fixed 0.00 (-0.01 to 0.01) 1.00 0% (1.00) NA Not effective Mortality Yang [191] 1 [192] 4 91 (0)/ 451 (15) 284 (0)/930 (15) OR Random 1.23 (0.38 to 3.97) 0.73 88% (<0.0001) N Not effective Das [193] 0 8 268 (0)/ 2009 (0) 533 (0)/3671 (0) OR Random 0.87 (0.46 to 1.64) 0.66 92% (<0.00001) Y Not effective Thoguluva Chandrasekar [194] 0 4 452 (0)/ 2111 (0) 125 (0)/1041 (0) OR Random 1.86 (1.38 to 2.50) <0.00 1 29% (0.234) NA Harmful Zang [195] 0 3 63 (0)/311 (0) 27 (0)/268 (0) RR Fixed 1.92 (1.26 to 2.93) 0.003 0% (0.508) NA Harmful Deterioration† Yang [191] 3 [55, 180, 192] 3 48 (2)/ 494 (116) 29 (4)/540 (126) OR Random 2.46 (0.42 to 14.45) 0.32 69% (0.007) N Not effective Liu [43] 3 [55, 180, 192] 0 2 (2)/ 115 (115) 4 (4)/125 (125) RR Random 0.96 (0.10 to 9.66) 0.98 41% (0.8) NA Not effective Wang [42] 2 [55, 192] 3 244 (1)/ 843 (46) 858 (4)/ 4112 (46) RR Random 1.05 (0.61 to 1.81) NA 62.5% (0.031) N Not effective Clinical progression within 5–7 days‡ Elsawah [190] 2 [55, 192] 2 11 (1)/89 (46) 6 (4)/83 (46) RD Fixed 0.06 (-0.03 to 0.15) 0.18 76% (0.006) NA Not effective Clinical progression within 28 days‡ Elsawah [190] 2 [179, 180] 0 9 (9)/ 206 (206) 11 (11)/ 234 (234) RD Fixed -0.00 (-0.04 to 0.04) 0.86 0% (0.33) NA Not effective Death or invasive MV Putman [196] 0 2 166 (0)/ 895 (0) 83 (0)/654 (0) HR Random 1.03 (0.82 to 1.29) 0.81 0% (0.75) NA Not effective Death or deterioration† Sarma [197] 2 [55, 192] 1 5 (1)/66 (46) 4 (4)/62 (46) OR Random 1.37 (0.09 to 21.97) 0.82 59% (0.09) NA Not effective Death or deterioration† (≤ 400 mg/day) Yang [191] 2 [55, 192] 2 64 (1)/365 (46) 270 (4)/787 (46) OR Random 0.64 (0.14 to 2.81) 0.55 84% (0.0002) N Not effective Death or deterioration† (> 400 mg/day) Yang [191] 1 [180] 1 5 (1)/90 (70) 0 (0)/96 (80) OR Fixed 6.17 (0.71 to 53.47) 0.10 0% (0.67) N Not effective Clinical recovery Misra [22] 2 [180, 192] 5 1026 (69)/ 1474 (90) 1054 (67)/ 1376 (90) RR Random 0.93 (0.84 to 1.04) NA 74% (<0.01) Y Not effective Talaie [198] 2 [55, 180] 0 70 (70)/ 106 (106) 67 (67 )/ 106 (106) RR Random 1.04 (0.85 to 1.28) NA 79.3% (0.028) NA Not effective Radiological improvement Ullah [199] 2 [55, 192] 1 40 (30)/ 56 (46) 33 (24)/58 (46) OR Random 1.98 (0.47 to 8.36) 0.36 54% (0.11) N Not effective Radiological progression Sarma [197] 2 [55, 192] 0 7 (7)/46 (46) 16 (16)/46 (46) OR Random 0.31 (0.11 to 0.90) 0.03 16% (0.27) NA Not effective Viral clearance Singh [200] 2 [180, 192] 1 80 (72)/ 99 (85) 81 (79)/111 (95) RR Random 1.05 (0.79 to 1.38) 0.744 62% (0.07) Y Inconclusive Liu [43] 2 [180, 192] 0 77 (77)/ 90 (90) 80 (80)/90 (90) RR Random 0.98 (0.89 to 1.07) 0.65 0% (0.54) NA Inconclusive Elavarasi [201] 0 3 217 (0)/ 240 (0) 152 (0)/203 (0) RR Random 1.21 (0.64 to 2.29) 0.56 87% (0.0006) NA Inconclusive Adverse events Wang [42] 3 [55, 180, 192] 1 35 (27)/ 200 (116) 10 (10)/223 (126) RR Fixed 3.62 (1.93 to 6.79) NA 17.6% (0.303) N Possible harm Zhong [189] 3 [55, 180, 192] 0 27 (27)/ 116 (116) 10 (10)/126 (126) RR Random 2.75 (1.42 to 5.33) NA 0% (0.442) NA Possible harm Adverse events (gastro- intestinal) Elsawah [190] 3 [179, 180, 192] 0 157 (157)/ 254 (254) 7 (7)/279 (279) RD Fixed 0.59 (0.55 to 0.64) <0.00 001 99% (<0.00001) NA Possible harm Adverse events (CNS) Elsawah [190] 3 [55, 179, 180] 0 65 (65)/ 270 (270) 3 (3)/295 (295) RD Fixed 0.23 (0.18 to 0.28) <0.00 001 99% (<0.00001) NA Possible harm Adverse events (neurological) Ullah [199] 2 [180, 192] 1 2 (2)/ 111 (101) 2 (0)/123 (111) OR Random 1.26 (0.20 to 7.98) 0.81 0% (0.37) N Inconclusive d l h [ ] [ ] ( )/ ( )/ ( ) d ( ) % ( ) l http://www.thno.org Theranostics 2021, Vol. Ribavirin Ribavirin has been investigated in previous studies on SARS and MERS, but the results were not consistent (Table 1). Although one RCT for a combination therapy of ribavirin and lopinavir/ ritonavir in SARS was registered [18], it seems unlikely that this trial can be finished, as SARS has not http://www.thno.org http://www.thno.org 1220 Theranostics 2021, Vol. 11, Issue 3 COVID-19 involving a total of 815 participants, treatment with hydroxychloroquine did not show better outcome compared to standard treatment (Table 3). In 15 meta-analyses on COVID-19, treatment with hydroxychloroquine showed no therapeutic effect and higher risk for adverse events. A combination of hydroxychloroquine and azithromycin was also evaluated in three meta-analyses on non-RCTs for COVID-19 and showed a harmful effect (Table 4). occurred for a while. In two in vitro studies on COVID-19, ribavirin did not show therapeutic effects [29, 30] (Table S1). In the aforementioned RCT on COVID-19, comparing “ribavirin plus interferon IFN-α with or without lopinavir/ritonavir” with “lopinavir/ritonavir plus IFN-α”, treatment with ribavirin did not show better outcome in terms of the clinical deterioration or viral clearance [41]. In another RCT on COVID-19, a combination of ribavirin and IFN-β1b was evaluated: In the treatment group, the time taken to achieve a National Early Warning Score 2 (NEWS2) (4 [IQR 3–8] vs. 8 [7–9] days; HR 3.92, 95% CI 1.66 to 9.23) or sequential organ failure assessment (SOFA) score of zero (3.0 [IQR 1.0–8.0] vs. 8.0 [6.5–9.0] days; HR 1.89, 95% CI 1.03 to 3.49), the hospitalization period (9.0 [IQR 7.0–13.0] vs. 14.5 [9.3–16.0] days; HR 2.72, 95% CI 1.2 to 6.13), and time to viral clearance (7 [IQR 5–11] vs. 12 [8–15] days; HR 4.37, 95% CI 1.86 to 10.24, P = 0.001) were shorter than those of the control group [44] (Table 3). Other antiviral agents In an RCT targeting SARS, early administration (within 7 days) of corticosteroids was associated with higher subsequent plasma viral concentrations in the second and third week of the illness [59]. However, in this study, the severity of disease did not differ between the early corticosteroid treatment group and the control group. In addition, there was no significant difference in the median time for the virus to become undetectable in plasma between the early corticosteroid treatment group and the control group (12 vs. 8 days, P = 0.106). Therefore, it is difficult to conclude that this study supports the risk of corticosteroid treatment. A non-RCT on SARS demonstrated that corticosteroid therapy was associated with higher risk for either ICU admission or mortality (OR 20.7, 95% CI 1.3-338.0) [60]. This study had several important limitations, including the following: (1) the 95% CI was extremely asymmetric; (2) there was no difference in mortality between the steroid-treated and non-treated groups in a simple univariate analysis, but corticosteroid therapy was included in the logistic regression; (3) the steroid-treated group had a more severe disease course, which indicates a case of confounding by indication; and (4) not all of the potential variables were adjusted, which could influence the results. In a small-scale RCT on mild to moderate COVID-19, azvudine (FNC) treatment showed better outcome with respect to radiological improvement (P = 0.0401) and viral clearance (P = 0.0011) [45]. In the other RCTs on COVID-19, triazavirin [46] or a combination of darunavir and cobicistat [47] did not show therapeutic effects (Table 3). Nelfinavir showed therapeutic effects in one [48] out of two in vitro studies on SARS (Table 1) and in an in vitro study on COVID-19 [49] (Table 2). Azithromycin In an RCT on azithromycin for COVID-19, the hospitalization period of the treatment group was shorter than that of the control group (4.6 [SD 2.6] vs. 6.0 [SD 3.2] days, P = 0.02) [57]. However, in another RCT on azithromycin involving 397 patients with severe COVID-19, azithromycin did not show any therapeutic effect [58] (Table 3). 4-Aminoquinoline In a study on SARS, chloroquine showed an effect in vitro but not in vivo, and these results were similar for amodiaquine [50]. The results of two in vitro studies of chloroquine for MERS conflicted with each other [51, 52]. Multiple in vitro studies on COVID-19 reported effects of chloroquine [29, 53, 54] and hydroxychloroquine [53, 54]. In a preprint RCT on COVID-19, hydroxychloroquine was administered with a daily dosage of 400 mg for five consecutive days and the treatment group showed higher rates of improvement in chest computed tomography (CT) scans on day 6 (25 out of 31 [81%] vs. 17 out of 31 [55%], P = 0.0476) and shorter duration of fever (2.2 [standard deviation (SD) 0.4] vs. 3.2 [1.3] days, P = 0.0008) [55]. In another preprint RCT conducted in Pakistan enrolling 500 patients with mild COVID-19, the proportion of patients with negative viral PCR results within seven days was higher in the hydroxychloroquine-treated group (182 out of 349 [52%] vs. 54 out of 151 [36%], P = 0.001) [56]. However, in the other four RCTs and one preprint RCT on In a non-RCT on MERS, corticosteroid therapy was not associated with 90-day mortality but associated with delay in viral clearance (adjusted HR 0.35, 95% CI 0.17-0.72) under a marginal structural model [61]. However, this study also shared many of the shortcomings mentioned above such as corticosteroids being used for patients with severe conditions, which can introduce severe levels of bias. http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 Theranostics 2021, Vol. 11, Issue 3 1221 In this retrospective study, at least a propensity-score matching analysis should have been considered. therapeutic anticoagulant therapy with enoxaparin and prophylactic anticoagulant therapy with enoxaparin or unfractionated heparin were compared. A greater proportion of participants in the therapeutic anticoagulant group were able to be weaned from MV successfully compared to the prophylactic anticoagulant group (8 out of 10 [80%] vs. 3 out of 10 [30%]; HR 4.0, 95% CI 1.04 to 15.05; P = 0.031) [75] (Table 3). In two meta-analyses including non-RCTs on COVID-19, anticoagulant therapy did not show a therapeutic effect [76, 77] (Table 4). Three non-RCTs of corticosteroid use in SARS showed effectiveness of high dose [62-64]. ACEI or ARB In a preprint RCT on 82 participants with COVID-19, telmisartan was administered to the treatment group with a daily dosage of 160 mg for 14 consecutive days. The treatment group had a shorter duration of hospital stay (9 vs. 15 days, P = 0.0124) and the HR for hospital discharge was 2.02 (95% CI 1.14 to 3.59) [72] (Table 3). In a meta-analysis that included three non-RCTs on COVID-19 with hypertension, ARB showed a survival benefit (OR for mortality 0.51, 95% CI 0.29 to 0.90, P = 0.02; I2 = 22%, n = 484) although there was a publication bias [73]. In another meta-analysis that included non-RCTs on COVID-19, the effect of ACEI or ARB therapy on COVID-19 with hypertension was not significant [74] (Table 4). Interferon Both IFN-α and IFN-β showed numerous positive results in non-human studies on SARS or MERS. However, a meta-analysis on MERS did not show that the interferon therapy was effective [14]. For COVID-19, five RCTs on interferon have been published to date. In an RCT evaluating IFN-β1a Calcifediol Calcifediol was studied in one RCT on COVID-19. In this RCT, a lower proportion of participants in the treatment group were admitted to the ICU compared to the control group (1 out of 50 [2%] vs. 13 out of 26 [50%]; adjusted OR 0.03, 95% CI 0.003 to 0.25) [78] (Table 3). Janus-associated kinase inhibitors In an in vitro study on MERS, baricitinib showed an effect [80] (Table 1). Ruxolitinib was evaluated in an RCT on severe COVID-19 and showed higher rates of improvement on chest CT scans on day 14 (18 out of 20 [90%] vs. 13 out of 21 [62%], P = 0.0495) and shorter time to lymphocyte recovery (5 [IQR 2–7] vs. 8 [2–11] days, P = 0.033) [81] (Table 3). CM4620-IE (AuxoraTM, calcium release- activated calcium channel inhibitor) In two RCTs on COVID-19, colchicine showed effects in major outcomes. Among them, in a Greek RCT, the treatment group had a higher cumulative event-free 10-day survival rate (97% vs. 83%, P = 0.03), a longer event-free survival period (21 [SD 0.31] vs. 19 [0.83] days, P = 0.03), and a lower incidence of deterioration within three weeks (2% vs. 14%; OR 0.11, 95% CI 0.01 to 0.96; P = 0.046) [70]. In another preprint RCT conducted in Brazil, the treatment group had a shorter duration of supplemental oxygen therapy (3.0 [IQR 1.5–6.5] vs. 7.0 [3.0–8.5] days, P = 0.02), a lower proportion of participants requiring supplemental oxygen on day 7 (6% vs. 39%, P = 0.01), a shorter length of hospital stay (6.0 [IQR 4.0–8.5] vs. 8.5 [5.5– 11.0] days, P = 0.03), and lower rate of hospitalization (53% vs. 78% on day 5; 6% vs. 17% on day 10; P = 0.01) [71]. In an RCT on severe or critical COVID-19, the proportion of patients who received invasive MV or died was lower in the Auxora-treated group than the control group (3 out of 17 [18%] vs. 5 out of 9 [6%]; HR 0.23, 95% CI 0.05 to 0.96; P < 0.05) and the mean difference in the 8-point ordinal scale was statistically significant on day 6 and day 9 to 12 (P < 0.05) [79] (Table 3). 4-Aminoquinoline One non-RCT on SARS demonstrated that the survival outcome of the group receiving methylprednisolone was superior compared to the group not receiving corticosteroids as well as the group receiving hydrocortisone or pulse therapy [65]. In an RCT on methylprednisolone treatment for severe COVID-19, the treatment group showed a lower mortality rate (2 out of 34 [6%] vs. 12 out of 28 [43%], P < 0.001) and a higher rate of clinical improvement (32 out of 34 [94%] vs. 16 out of 28 [57%], P = 0.001) compared to the control group [66]. Meta-analyses on corticosteroid therapy for COVID-19 included only non-RCTs and did not demonstrate any significant therapeutic effect of corticosteroids [67-69] (Table 4). Leflunomide (dihydroorotate dehydrogenase inhibitor) In a small-sized RCT on moderate COVID-19, the duration of viral shedding was shorter in the leflunomide-treated group compared with the control group (5 vs. 11 days, P = 0.046) [82] (Table 3). Discussion We summarized the results of studies conducted on SARS, MERS, and COVID-19 to date. Unfortunately, completed RCTs for the treatment of SARS and MERS were scarce. We assumed this was because SARS was a relatively short-lived epidemic that has not occurred since 2004, and the number of patients with MERS might have been insufficient for recruitment. In the case of COVID-19, numerous RCTs have been registered, and research results have been consistently reported despite the global pandemic and medical crisis. It was difficult to find an optimal therapeutic agent that consistently resulted in positive outcomes across SARS, MERS, and COVID-19. One of the possible reasons of this is that there might not be a universal “cure” to these viral diseases given the differences in presentation forms. Reduction of the viral load may not be the only aim when attempting to cure the disease. The subtle differences between these three coronaviruses, as well as the lack of objective information from clinical experiences of the preceding SARS and MERS epidemics, may also be Other immunotherapies In a small-scale RCT on severe COVID-19, vilobelimab (anti-C5a antibody IFX-1) treatment did not show a therapeutic effect [92]. In a small-scale RCT on CIGB (anti-CK2) for COVID-19, there was a reduction in the number of pulmonary lesions on chest CT in a greater proportion of participants in the treatment group compared to the control group (5 out of 6 [83%] vs.3 out of 7 [43%]; Bayesian P (difference > 0) = 0.951) [93]. Multiple observational clinical studies on tocilizumab (anti-interleukin [IL] -6 receptor antibody) for COVID-19 were investigated in two meta-analyses. Among them, a subgroup analysis, in which lopinavir and ritonavir were and corticosteroids were not administered to all participants, showed a lower mortality rate in the tocilizmub treatment group (risk difference -0.31, 95% CI -0.57 to -0.05) [94] (Table 4). Convalescent plasma A meta-analysis of convalescent plasma therapy in patients with SARS demonstrated that the absolute reduction in the risk of mortality was 7% and 23% in two studies, and the case fatality rate from four non- comparative studies varied from 0% to 12.5% [12]. In an RCT for severe or life-threatening COVID-19, the proportion of participants who recovered clinically within 28 days was higher in the treatment than in the control group in cases with severe COVID-19 (21 out of 23 [91%] vs. 15 out of 22 [28%]; HR 2.15, 95% CI 1.07 to 4.32) [87]. Four preprint RCTs on COVID-19 have investigated the effect of convalescent plasma. In one of these RCTs, a lower proportion of participants in the treatment group compared to the control group either required MV or died (0 out of 38 [0%] vs. 6 out of 43 [14%], P = 0.03) [88]. Another RCT on moderate COVID-19 involving more participants (n = 464) did not show the therapeutic effect of convalescent plasma [89] (Table 3). Anticoagulants In a small-scale RCT for severe COVID-19, In a small-scale RCT for severe COVID-19, http://www.thno.org 1222 Theranostics 2021, Vol. 11, Issue 3 treatment, the rate of hospital discharge was higher (67% vs. 44%; OR 2.5, 95% CI 1.05 to 6.37) and the 28-day mortality rate was lower (19% vs. 44%, P = 0.015) in the treatment group compared to the control group [83]. In the aforementioned RCT evaluating a combination of IFN-β1b and ribavirin, the treatment group performed better [44]. In another RCT on severe COVID-19, treatment with IFN-β1b showed better outcomes in terms of discharge from the hospital and admission to the ICU compared to standard treatment [84]. Inhaled IFN-κ plus TFF2 therapy was investigated in one RCT on moderate COVID-19. In this study, the chest CT findings of participants in the treatment group improved within a shorter time compared to the control group (6.2 [95% CI 5.1–7.3] vs. 8.8 [95% CI 7.6-10.0] days, P = 0.002) [85]. In an RCT evaluating Novaferon therapy, the group receiving the combination of Novaferon and lopinavir/ritonavir had a higher rate of viral clearance on day 6 (18 out of 30 [60%] vs. 7 out of 29 [24%], P = 0.0053) and a shorter median time to negative results of virus PCR (6 vs. 9 days, P = 0.036) compared with the group receiving lopinavir/ ritonavir [86] (Table 3). Intravenous immunoglobulin In a preprint RCT on COVID-19, the 3-day course of intravenous immunoglobulin therapy showed a lower rate of MV within 30 days (2 out of 14 [14%] vs. 7 out of 12 [58%], P = 0.038), shorter hospital stay (11 [range 5–22] vs. 19 [4–30] days, P = 0.013) or ICU stay (2.5 [range 0–16] vs. 12.5 [1–29] days, P = 0.006), and improvement in PaO2/FiO2 ratio on day 7 (difference +131 [+35 to +330] vs. +44.5 [-115 to +157], P = 0.01) among the participants who had an alveolar-arterial oxygen gradient greater than 200 mmHg at enrollment [91] (Table 3). other reasons. the RNA dependent RNA polymerase (RdRp) of RNA viruses [97]. Considering that favipiravir showed an effect in terms of viral clearance in an RCT on COVID-19 [31], a large-scale, well-designed RCT is further required. A number of RCTs on favipiravir are now underway. Synthesizing studies on COVID-19 highlighted two main goals in the treatment of COVID-19: (1) effective elimination of the virus and (2) immune regulation to interfere with the mechanisms of cytokine storm. Therefore, extensive further research on various antiviral agents and immunomodulators is expected to continue for a while. Umifenovir is a small indole-derivative molecule with broad-spectrum antiviral property, and it has been approved in Russia and China for the prophylaxis and treatment of influenza. It inhibits the virus from fusion to the target cell membrane and blocks viral entry into the cell [98]. Since a meta-analysis including one RCT and multiple observational studies on umifenovir for COVID-19 demonstrated significant results for viral clearance [35], well-designed RCTs on umifenovir still needs to be conducted. p Among the antiviral agents presented in our study, remdesivir has consistently shown potent effects in non-human studies on SARS, MERS, and COVID-19. Remdesivir is a novel broad-spectrum antiviral agent. When remdesivir is administered into the human body, it is metabolized to an active metabolite, which is an adenosine nucleoside triphosphate analogue. It interferes with the action of viral RNA polymerase and evades proofreading by viral exoribonuclease (ExoN), which interferes with RNA replication of the virus [95]. This agent showed an effect in the largest of the four RCTs on COVID-19 that included 1,062 participants. In this RCT, the 14-day mortality rate was significantly lower in the subgroup with requirement of only supplemental oxygen not MV, suggesting that there is a patient population with a certain level of severity of COVID-19 that could benefit from remdesivir treatment [20]. Therefore, it seems necessary to preferentially administer remdesivir to this specific group of patients. In addition, it should be considered in the clinical application that the outcome of 5-day regimen treatment was better than that of the 10-day regimen in two RCTs [23, 24]. On October 22, 2020, the Food and Drug Administration (FDA) approved the use of remdesivir for treatment of COVID-19 requiring hospitalization. Lopinavir/ritonavir combination therapy seems to have more therapeutic effect than monotherapy of each drug. other reasons. As protease inhibitors, lopinavir and ritonavir can inhibit the action of 3CLpro, coronavirus main protease, and interfere with the process of viral replication and release [99]. However, the clinical results of this combination therapy for COVID-19 were not meeting expectations compared to those of SARS and MERS. In a first RCT on lopinavir/ritonavir for COVID-19, the outcomes of the treatment group and the control group did not show a significant difference, but the 28-day mortality of the treatment group were slightly lower [40]; and in two other RCTs and four meta-analyses on COVID-19, treatment with lopinavir/ritonavir did not show an effect. Azvudine (FNC) is a novel nucleoside reverse transcriptase inhibitor targeting HCV and has been investigated for treatment of human immuno- deficiency virus (HIV) [100]. Azvudine showed promising effects in terms of radiological improvement and viral clearance in a small-scale RCT for mild to moderate COVID-19 [45]. RCTs involving more participants are required. A combination of sofosbuvir and daclatasvir consistently showed an effect on the major outcomes in two RCTs on COVID-19. Sofosbuvir is a nucleotide analog targeting the hepatitis C virus (HCV) polymerase, NS5B. This agent is capable of inhibition of positive-strand RNA viruses like coronavirus. Daclatasvir is an HCV NS5A antagonist and is known to penetrate lung tissue effectively. Daclatasvir has been shown to inhibit the production of SARS-CoV-2 particles in an in vitro study [96]. Those two RCTs investigating the combination of sofosbuvir and daclatasvir had limitations because they included a small number of participants and did not show significant results for mortality. Therefore, a large- scale RCT on this treatment is still required. The 4-aminoquinoline is mainly used as an anti-malaria agent, and hydroxychloroquine, one of its derivatives, is also used as an immunomodulatory agent. Chloroquine and hydroxychloroquine inhibits the pH-dependent steps of viral replication by increasing the pH of phagolysosome [101]. The study on SARS showed in vitro but not in vivo effects of chloroquine and amodiaquine [50], and combination therapy with other drugs that inhibit viral replication may be necessary. In our study, we investigated seven RCTs and 15 meta-analyses on hydroxychloroquine for COVID-19 and only one preprint small-scale RCT demonstrated an effect of hydroxychloroquine on the major outcomes [55]. The survival benefit of hydroxychloroquine treatment has not been Favipiravir is an antiviral agent that is drawing attention as a treatment option for COVID-19. Theranostics 2021, Vol. 11, Issue 3 1223 Recombinant human granulocyte colony-stimulating factor (rhG-CSF) In an RCT for COVID-19 with lymphopenia, participants in the treatment group who received rhG-CSF showed lower rates of 21-day mortality (2 out of 100 [2%] vs. 10 out of 100 [10%]; HR 0.19, 95% CI 0.04 to 0.88) and disease progression (2 out of 100 [2%] vs. 15 out of 100 [15%]; mean difference -13%, 95% CI -21.4% to -5.4%) [90] (Table 3). http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 other reasons. It is promising that the effects of colchicine treatment were revealed in terms of survival, clinical improvement, and duration of hospitalization in the two RCTs for COVID-19 [70, 71]. If a larger-scale follow-up RCT is conducted, the effect might be further supported. For severe inflammatory diseases caused by infection, corticosteroid therapy is a double-edged sword. Although a number of corticosteroid therapies have already been used in SARS, MERS, and COVID-19, these results are controversial and difficult to interpret. It is noteworthy that the study of SARS showed a difference in outcomes depending on the type or dosage of steroids [65]. In most observational studies on COVID-19, corticosteroid therapy was mainly administered in a group with severe clinical conditions according to the prevailing guidelines [7]. Because of this strong tendency, patients with COVID-19 who received corticosteroids had poor treatment outcomes, and objective validation of corticosteroid treatment has been highly required. pp Angiotensin-converting enzyme 2 (ACE2) is a transmembrane protein and the main entry point into cells for SARS-CoV-2. Theoretically, if the expression of ACE2 decreases, it will be a defense mechanism against the entry of the virus. On the other hand, ACE2 shows a protective action against virus-induce lung injury by converting angiotensin II to angiotensin-(1–7), which have a vasodilator effect [110, 111]. ACEI and ARB can induce up-regulation of ACE2 [112, 113], which might negatively affect the treatment of disease. Contrary, an in vivo study showed that SARS-CoV spike-mediated lung injury was attenuated by losartan [114]. For these conflicting evidences, there has been an interest in ACEI and ARB in relation to COVID-19. In one RCT for COVID-19 [72] and one meta-analysis including three non-RCTs on COVID-19 [73], participants who received ARB performed better in terms of discharge or survival. More RCTs on ACEI or ARB for COVID-19 in the group of patients with pre-existing hypertension or at risk for cardiovascular disease are still required. In response to this request, the results of the first RCT to investigate corticosteroid therapy in COVID-19 were recently reported, although it was excluded from our study because participants with negative SARS-CoV-2 PCR results were included. This RCT involved 2,104 and 4,321 participants in the treatment and control group, respectively. The 28-day mortality rate of the group receiving 10 days of dexamethasone treatment was lower than that of the control group (482 out of 2,104 [23%] vs. other reasons. The active metabolite of favipiravir competes with purine nucleosides and incorporates into viral RNA to interfere with viral replication, potentially inhibiting Theranostics 2021, Vol. 11, Issue 3 1224 demonstrated in any of these seven RCTs. The results of the latest large-scale RCT [102], which was excluded from our study since it included patients with suspected COVID-19, as well as multiple meta-analyses were consistent with this. In addition, the risk for adverse events of hydroxychloroquine treatment was higher compared to standard treatment according to the results of meta-analyses. Therefore, hydroxychloroquine seems to have no value as a therapeutic agent for COVID-19. (29%) in the treatment group and 283 out of 683 patients (41%) in the control group among patients requiring invasive MV (RR 0.64, 95% CI 0.51 to 0.81) (2) 298 out of 1,279 patients (23%) in the treatment group and 68 out of 2,604 patients (26%) in the control group among patients requiring only oxygen supplement (RR 0.82, 95% CI 0.72 to 0.94). There was no significant difference in the 28-day mortality rate among patients not receiving respiratory support [108]. These results strongly suggest that dexamethasone treatment is effective for a population of patients with COVID-19 requiring respiratory support. It is necessary to confirm these results among the participants who were diagnosed as COVID-19 through viral RNA detection. In another RCT for severe COVID-19, patients receiving methyl- prednisolone also showed a lower mortality rate compared to the control group [66]. It is worth noting that an RCT for chloroquine, excluded due to involvement of patients with suspected COVID-19 [103], suggested the risk of high-dose chloroquine treatment and this risk was associated with prolongation of the QT interval. Similarly, hydroxychloroquine treatment was also related to QT prolongation [104, 105]. Therefore, it is expected that if careful monitoring for prolongation of the QT interval is not accompanied, chloroquine or hydroxychloroquine treatment can be rather harmful. In this regard, the World Health Organization (WHO) recently decided to implement the temporary pause of the hydroxychloroquine arm within the Solidarity Trial, a large-scale study on four untested treatments for COVID-19 [106]. The FDA also withdrew emergency use authorization for chloroquine and hydroxychloroquine [107]. Colchicine, an anti-inflammatory agent which is mainly used for gout and rheumatoid arthritis, has been used for a long time. It inhibits microtubule polymerization and mitosis in the metaphase [109]. other reasons. In one RCT for severe COVID-19, convalescent plasma treatment showed an effect in terms of clinical improvement [87]. In contrast, the effect of convalescent plasma treatment was not demonstrated in another RCT on moderate COVID-19 involving larger number of participants [89]. The differences in severity of COVID-19 in these two RCTs may have contributed to this contradiction. Therefore, a large-scale RCT on convalescent plasma treatment targeting severe COVID-19 is required. CM4620-IE (AuxoraTM) is a selective small molecule inhibitor of calcium release-activated calcium (CRAC) channels. It was developed to prevent over-activation of CRAC channels that can lead to inflammatory diseases. It has been suggested that Auxora may protect against pulmonary endothelial damage and cytokine storm [119, 120]. This agent has been shown to be effective in terms of survival and clinical improvement in one small-scale RCT for severe or critical COVID-19 [79]. Ruxolitinib is a potent selective inhibitor of Janus-associated kinases 1 and 2 and has been used as a treatment for primary myelofibrosis, post- polycythemia vera or post-essential thrombocythemia myelofibrosis [121]. Ruxolitinib has a broad-spectrum of anti-inflammatory properties against cytokine storm mediated by IL-1, IL-6, IL-8, IL-12, tumor necrosis factor-α, IFN-γ, vascular endothelial growth factor, and granulocyte-macrophage colony- stimulating factor [122]. In an RCT on severe COVID-19, ruxolitinib showed higher rate of radiological improvement despite the small number of participants [81]. In this RCT, no one out of 20 patients with severe COVID-19 in the treatment group died within 28 days, whereas three out of 21 patients in the control group died. Further research results for severe COVID-19 at high risk for cytokine storm need to be supplemented. q Intravenous immunoglobulin therapy provides passive immunity and has the property to modulate immune function. High doses of intravenous immunoglobulin can produce anti-inflammatory and inflammatory-modulating effects on a variety of immune cells, which can intervene and modulate the mechanisms of cytokine storm, and have been administered to treat various diseases such as immune thrombocytopenia purpura or Kawasaki disease [126]. The only preprint small-scale RCT on intravenous immunoglobulin therapy for COVID-19 showed better clinical outcomes in the treatment group [91]. Leflunomide, an isoxazole derivative, inhibits the T cell proliferation by blocking dihydroorotate dehydrogenase. This agent has been used in the treatment of rheumatoid arthritis and psoriatic arthritis, and has been attempted to treat BK virus, cytomegalovirus, HIV, and ebolavirus [123]. other reasons. 1,110 out of 4,321 [26%]; RR 0.83, 95% CI 0.75 to 0.93; P < 0.001), and this trend was stronger among the subgroup with higher severity of COVID-19: (1) 95 out of 324 patients Routine administration of anticoagulants in sepsis or ARDS is not recommended currently. However, disseminated intravascular coagulation should still be the target of research to find treatments for sepsis or ARDS, because it is deeply involved in http://www.thno.org 1225 Theranostics 2021, Vol. 11, Issue 3 the pathogenesis and progress of these diseases. It has been reported that coagulopathy was associated with the prognosis of COVID-19 [115-117] and these results are consistent with what has been known in ARDS and sepsis. In a small-scale RCT of severe COVID-19, therapeutic anticoagulant therapy with enoxaparin showed a better outcome than prophylactic anticoagulant therapy [75]. More studies investigating the efficacy of augmenting these anticoagulation or thrombolytic treatments, while weighing the risk of hemorrhage, and narrowing the indications are required. a preprint observational study involving 27 participants, leflunomide showed effects of promoting viral clearance and increasing discharge rate [124]. Interferon showed effects in a number of in vitro studies on SARS and MERS. In addition, three RCTs on IFN-β for COVID-19 demonstrated favorable results in terms of survival, clinical improvement, discharge from hospital, and viral clearance [44, 83, 84]. Since interferon has been used as a combination therapy with antiviral agents in most cases, further research is needed to discover the antiviral agent that can show the greatest effect when administered in combination with interferon, as well as specific indications. Calcifediol is a main metabolite of vitamin D. Since lung epithelium expresses vitamin D receptors, administration of vitamin D may suppress the development of ARDS [118]. In a pilot RCT on COVID-19, treatment with a high dosage of calcifediol reduced the need for ICU admission [78]. Convalescent plasma contains pathogen-specific neutralizing antibodies that can neutralize viral particles, which provide passive immunity to the recipient. It is hypothesized that early convalescent plasma therapy enhances the patient's capability to clear the initial viral inoculum by neutralizing viral particles [125]. Convalescent plasma therapy has been applied to a wide range of infectious diseases such as diphtheria, pneumococcal pneumonia, hepatitis A and B, mumps, polio, measles, and rabies. The results of a meta-analysis on convalescent plasma treatment for SARS are relatively promising [12]. other reasons. The only RCT evaluating leflunomide for COVID-19 enrolled only 10 participants, but showed an effect of shortening the viral shedding period [82]. Similarly, in Our study has some limitations. RCTs for SARS and MERS were extremely rare. In the case of COVID-19, more RCTs were obtained. However, all except for eight RCTs included less than 100 participants per each arm. Although we updated the http://www.thno.org Theranostics 2021, Vol. 11, Issue 3 Theranostics 2021, Vol. 11, Issue 3 1226 latest search results for RCTs and meta-analyses on COVID-19, we did not include the latest search results for non-clinical studies on the three coronavirus diseases because of the vast amount of data. distress syndrome; CI: confidence interval; COVID-19: coronavirus disease 2019; CRAC: calcium release-activated calcium; CT: computed tomography; EC50 : 50% maximal effective concentration; ExoN: exoribonuclease; FDA: Food and Drug Administration; HCV: hepatitis C virus; HIV: human immunodeficiency virus; HR: hazard ratio; ICU: intensive care unit; IFN: interferon; IL: interleukin; IQR: interquartile range; MERS: Middle East respiratory syndrome; MV: mechanical ventilation; NEWS2: National Early Warning Score 2; OR: odds ratio; PCR: polymerase chain reaction; PICO: Participants, Interventions, Comparisons, and Outcomes; PRISMA: Preferred Reporting Items for Systematic Reviews and Meta-Analyses; RCT: randomized controlled trial; RdRp: RNA dependent RNA polymerase; rhG-CSF: recombinant human granulocyte colony-stimulating factor; RR: relative risk; SARS: severe acute respiratory syndrome; SARS-CoV-2: severe acute respiratory syndrome coronavirus 2; SD: standard deviation; SI: selectivity index; SOFA: sequential organ failure assessment; WHO: World Health Organization. In addition, our study also highlights that treatments with potential effects seen in in vitro studies have not translated in positive in vivo or clinical studies. The 4-aminoquinoline derivatives showed effects in a number of in vitro studies, but not in in vivo and clinical studies. On the other hand, favipiravir showed unfavorable results in an in vitro study, while it showed effects in a clinical study. This contradiction between in vitro and in vivo studies or between pre-clinical and clinical studies does not help in the current situation where a therapeutic agent for COVID-19 must be discovered in a short amount of time. Therefore, it is important to design in vivo or clinical studies after a thorough understanding of drug pharmacology and in-depth consideration of how to link in vitro antiviral activity and drug exposure at the putative target site of action. Fan et al. Author contributions In this summary report, we synthesized the results of previous studies on the treatment of SARS, MERS, and COVID-19. There was no therapeutic agent that consistently resulted in positive outcomes across SARS, MERS, and COVID-19. Remdesivir showed a therapeutic effect for COVID-19 in two RCTs involving the largest number of total participants (n = 1,461). Other therapies that showed an effect in at least two RCTs for COVID-19 were sofosbuvir/daclatasvir (n = 114), colchicine (n = 140), IFN-β1b (n = 193), and convalescent plasma therapy (n = 126). Further RCTs are required. YJH and JIS conceived and designed the Review. YJH and JIS selected the articles, extracted and analyzed the data, and wrote the first draft of the manuscript. YJH, KHL, SY, SWN, SR, DS, JSK, JYL, JWY, JHL, AKo, SHH, ED, JR, LS, HO, RAG, AKr, ME, DK, SD, WK, LJ, HS, and JIS interpreted the data and contributed to the writing of the final version of the manuscript. All authors critically revised the manuscript and agreed with the results and conclusions of this Review. Acknowledgements When a specific infection enters a pandemic state, group immunization through vaccine, rather than quarantine, is the most effective countermeasure. As of October 29, 2020, 201 candidate vaccines against SARS-CoV-2 are being developed. Among them, 45 have entered clinical trials, and none has been approved for use yet [129]. We appreciate Kyungsoo Park, Professor of Department of Pharmacology, Yonsei University College of Medicine, Seoul, Republic of Korea, and Jae Yong Chung, Professor of Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine, Seoul, Republic of Korea. They contributed to this study by giving valuable advice on interpreting in vitro studies. Supplementary Material Supplementary table S1. Supplementary table S1. http://www.thno.org/v11p1207s1.pdf Supplementary table S1. pp y http://www.thno.org/v11p1207s1.pdf other reasons. demonstrated that in vitro EC50/EC90 values for hydroxychloroquine should be compared to the in vivo free extracellular tissue concentration, which is similar to the free plasma hydroxychloroquine concentration [127]. Advances in cell modeling tools for biological research are expected to further enrich preclinical research design, and also help promote the development of new therapies [128]. References Momattin H, Al-Ali AY, Al-Tawfiq JA. A Systematic Review of therapeutic agents for the treatment of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV). Travel Med Infect Dis. 2019; 30: 9-18. g 37. Chan KS, Lai ST, Chu CM, Tsui E, Tam CY, Wong MM, et al. Treatment of severe acute respiratory syndrome with lopinavir/ritonavir: a multicentre retrospective matched cohort study. Hong Kong Med J. 2003; 9: 399-406. 14. 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https://openalex.org/W4311390627	https://iris.uniroma1.it/bitstream/11573/1677275/1/Basile_Treatment_2022.pdf	English	null	Treatment of Femoral Shaft Pseudarthrosis, Case Series and Medico-Legal Implications	Journal of clinical medicine	2,022	cc-by	7,349	Citation: Basile, G.; Fozzato, S.; Petrucci, Q.A.; Gallina, M.; Bianco Prevot, L.; Accetta, R.; Zaami, S. Treatment of Femoral Shaft Pseudarthrosis, Case Series and Medico-Legal Implications. J. Clin. Med. 2022, 11, 7407. https://doi.org/ 10.3390/jcm11247407 Academic Editor: Heinrich Resch Received: 30 October 2022 Accepted: 13 December 2022 Published: 14 December 2022 Citation: Basile, G.; Fozzato, S.; Petrucci, Q.A.; Gallina, M.; Bianco Prevot, L.; Accetta, R.; Zaami, S. Treatment of Femoral Shaft Pseudarthrosis, Case Series and Medico-Legal Implications. J. Clin. Med. 2022, 11, 7407. https://doi.org/ 10.3390/jcm11247407 Keywords: pseudarthrosis; non-union; femoral shaft fractures; medical professional liability; bone graft Journal of Clinical Medicine Journal of Clinical Medicine Journal of Clinical Medicine Journal of Clinical Medicine Giuseppe Basile 1,†, Stefania Fozzato 1,† , Quirino Alessandro Petrucci 2, Mario Gallina 1, Luca Bianco Prevot 1, Riccardo Accetta 1 and Simona Zaami 2,* Giuseppe Basile 1,†, Stefania Fozzato 1,† , Quirino Alessandro Petrucci 2, Mario Gallina 1, Luca Bianco Prevot 1, Riccardo Accetta 1 and Simona Zaami 2,* 1 IRCCS Orthopedic Institute Galeazzi, 20161 Milan, Italy 1 IRCCS Orthopedic Institute Galeazzi, 20161 Milan, Italy 2 Department of Anatomical, Histological, Forensic and Orthopedic Sciences, “Sapienza” University of Rome, 00161 Rome, Italy C d 2 Department of Anatomical, Histological, Forensic and Orthopedic Sciences, “Sapienza” University of Rome, 00161 Rome, Italy * Correspondence: simona.zaami@uniroma1.it † These authors contributed equally to this work. Abstract: Pseudarthrosis (PSA) is a possible complication of femoral shaft fracture treatment. It is often associated with reduced bone quality and can, therefore, adversely affect quality of life. Its treatment poses a major challenge for orthopaedic surgeons. Several authors have set forth different surgical approaches for the treatment of pseudarthrosis, such as internal ﬁxation with plate and screws, replacement of an intramedullary nail or prosthetic replacement. In cases associated with bone loss, osteopenia, or comminution of fracture fragments, autologous or homologous bone grafts may also be used. The chronic outcomes of the surgical treatment of femoral shaft pseudarthrosis, even when consolidation is achieved, are linked to disabling sequelae of clinical-functional relevance, deserving an adequate medico-legal evaluation. The purpose of this retrospective study is to analyse a clinical case series of patients treated for atrophic femoral shaft pseudarthrosis at the IRCCS Orthopaedic Institute Galeazzi, Milan, Italy, from 2014 to 2020 and their orthopaedic-traumatological and medico-legal implications. 1. Introduction In closed fractures, treated conservatively, a muscle ﬂap was sometimes interposed between the fragments, which constituted an insuperable barrier for the repairing tissue. Furthermore, it was believed that the non-consolidation was caused by an insufﬁcient period of immobilisation in the plaster cast and/or by premature weight-bearing loading and walking [15]. g In the past, femur PSA was believed to be mostly related to errors in the treatment method. Recent research has shown that non-union, both atrophic and hypertrophic, is related to a certain degree of vascularisation and mesenchymal stem cells (MSCs) with osteogenic capacity. The non-consolidation of a femur fracture causes pain, especially during loading, altered and excessive motility at the fracture focus, functional limitations of the contiguous joints and muscle hypotrophy. It can therefore cause serious damage to the “functioning” capabilities of the injured lower limb, with consequent qualitative and quantitative alter- ations of everyday life activities. The lack of bone continuity of the femoral shaft and the consequent preternatural motility with instability to biomechanical stress prevent lower limb loading and severely reduce active and passive mobility, as well as walking. Accord- ing to the disability model of the International Classiﬁcation of Functioning, Disability and Health (ICF), adopted by the World Health Organization (WHO), ﬁve levels of disability involving the lower limbs have been identiﬁed: level 0 interference with the function is not demonstrable; level 1 interference is associated with vigorous exercise; level 2 analgesic claudication is presented and limits the distance travelled or requires the constant and regular use of an orthosis or a cane; level 3 presence of analgesic claudication with constant use of two sticks associated with orthoses; level 4 is impossible to walk [16]. By adopting these criteria, femoral PSA causes level 3 or 4 disability, with severe or total functional impotence of the lower limb. To this damage are added the deformities caused by the frequent displacement of the fracture’s fragments, the dysmetria, the severe hypotonia from non-use, the joint stiffness of the knee and, to a lesser extent, of the hip [17]. In order to minimise complications, surgical and non-surgical treatment of femoral fracture must be adapted differently for each patient, considering the biological and biomechanical basis of bone healing. 1. Introduction Pseudarthrosis (PSA) has been deﬁned in different ways in the literature. It has been described as a lack of radiographic consolidation at six months after the traumatic event and is associated with the necessity for revision surgery. According to the Food and Drug Administration (FDA), the diagnosis of PSA is made after at least nine months from the trauma with no visible signs of progressive healing for three months [1,2]. A higher incidence of PSA has been observed in femoral shaft fractures in recent years. However, it is an infrequent complication compared with pseudarthrosis of the humeral shaft [1], whose incidence is approximately 1.9–10% of all fractures occurring annually [1] or compared with PSA in tibial fractures that are about 13.3% [3,4]. The increased incidence of PSA in femoral fractures, whose incidence is approximately 1.9–5% of all fractures occurring annually [1,5], is due to the increased survival of patients with multiple severe injuries and the broadening of indications for intramedullary nailing [6]. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional afﬁl- iations. Numerous techniques have been proposed for the treatment of femoral shaft pseu- darthrosis, including electromagnetic ﬁelds [7], low-intensity ultrasound [8], extracorporeal shock wave therapy (ESWT) [9], external ﬁxators [10] and osteosynthesis with plate and screws [11]. Moreover, several studies have reported positive results after the treatment of non-union of the femur with single or double plate ﬁxation combined with autologous bone graft [12,13]. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). Undoubtedly, the major severity of the conditions of road polytrauma patients and the increase in the average survival age have led to therapeutic problems due to the greater complexity of fractures and associated comorbidities such as obesity, diabetes, Chronic https://www.mdpi.com/journal/jcm J. Clin. Med. 2022, 11, 7407. https://doi.org/10.3390/jcm11247407 J. Clin. Med. 2022, 11, 7407 2 of 10 Obstructive Pulmonary Disease (COPD), correlated visceral injuries, and neurological diseases, including the pathological consequences of the recent severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) pandemic [14]. y p In the 1970–1980s, femoral shaft PSA was considered an exceptional complication, while delayed union was less rare, affecting open fractures, especially with bone loss and in- fection. 1. Introduction Over the past decade, the “diamond concept” for fracture healing has ascribed equal importance to mechanical stability and the biological environment, offering a new paradigm for complex fractures and the management of compromised joints [18,19]. This article has been conceived to elaborate on a series of clinical cases of atrophic pseu- darthrosis after femoral shaft fracture treated by our team and to highlight the possible medico-legal repercussions. In fact, the complexity of these injuries can severely impact the ergonomic efﬁciency of the entire limb, resulting in possible postural alterations, with all the relative repercussions on the overall compromised structure of the patient. 2. Materials and Methods This study is centred on 16 patients, both male and female, with atrophic PSA af- ter femoral shaft fracture, treated at the IRCCS Orthopaedic Institute Galeazzi, Milan, Italy, between 2014 to 2020. Patient charts, operation, and follow-up notes were retrospec- tively analysed. The indication for revision surgery was given in case of rupture of the ﬁxation tools, severe osteopenia, cortical bone defect of ≥2 cm at the site of PSA or in the presence of disabling clinical symptoms. Surgical treatment of atrophic PSA of the femoral stem requires complete removal of the interposed necrotic tissue, remodelling of bone fragments and the restoration of fracture stability. Plate osteosynthesis, applied with an axial compression mechanism, is J. Clin. Med. 2022, 11, 7407 3 of 10 3 of 10 the best choice for most surgeons. The application of cortico-spongy grafts can increase the likelihood of consolidation. The graft, harvested from the patients themselves (autologous) or from a donor (homologous), enhances the mechanical stability of the synthesis to ensure total immobility of the fragments; in some cases, two plates mounted in parallel can be used for the treatment of humeral shaft PSA, although it may determine increased soft tissue dissection and a higher risk of bleeding. Open reduction and internal ﬁxation with a long compression plate combined with homologous bone grafting is our treatment of choice, even in osteoporosis patients with reduced bone quality. However, in selected cases previously treated with an intramedullary nail, we have considered dynamisation of the nail when possible or replacement of the intramedullary nail after further reaming the femoral canal. Only in case of impossibility to perform a revision of the surgical synthesis has prosthetic replacement been considered. Healing after the second surgery was radiographically assessed through the presence of a callus in the PSA site with the restoration of bone continuity and through a physical and clinical examination to determine the solidity of consolidation. As preoperative antibiotic prophylaxis, cefazolin 2 g was administered intravenously, except for two patients who were given clindamycin 600 mg intravenously due to allergy concerns. Antibiotic prophylaxis was continued until the ﬁfth postoperative day. Given the attention to residual haemostasis at the end of the surgical revision, we never used surgical drains. The surgical wound dressing was renewed every two days. The rehabilitation of the lower limb muscles started with isometric exercises and passive mobilisation of the affected limb the day after the surgery. 2. Materials and Methods Some cases may even call for pharmacological interventions (e.g. the administration of anabolic steroids) [20,21]. ® An articulated knee brace with long splints (DonJoy X-Act ROM Knee Orthosis®, DJO global®, Lewisville, TX, USA) was placed after surgery in all the patients for thirty days. The 20% partial weight bearing started at four weeks after surgery. At eight weeks, in the presence of radiographic signs of healing, such as the appearance of normal osteo- reparative phenomena at the level of the pseudarthrosis focus, the patient was granted a tolerance load with two crutches. Until complete weight bearing was reached, the patient was advised to use vascular elastic stockings associated with the administration of low molecular weight heparin. All patients underwent periodic clinical and radiographic follow-ups. All data were collected through a retrospective review of patient medical records and radiographs, and a minimum follow-up of 6 months after deﬁnitive surgery was requested. Informed consent was obtained from all patients. The nature of the study did not require approval from the institute’s ethics committee. 3. Results All patients were treated by the same surgeon and the mean intervention time was 163 min (range 40–236). g Seven patients were treated with dedicated plate and screws (NCB® plate, Zimmer Inc., Warsaw, IN, USA) associated with homologous bone graft, using a standard lateral surgical approach to the femoral shaft. After the removal of the previous implants, the focus of pseudarthrosis was exposed, removing the ﬁbrous tissue between the bone fragments, and the bone stumps were reduced under ﬂuoroscopic control with a C-arm. A bone graft splint was applied as a medial buttress, and internal ﬁxation was performed with a plate and screw at the lateral side. The anatomical planes were then sutured before applying a sterile dressing (Figure 1). ( g ) Seven patients were treated with dedicated plate and screws (NCB® plate, Zimmer Inc., Warsaw, IN, USA) associated with homologous bone graft, using a standard lateral surgical approach to the femoral shaft. After the removal of the previous implants, the focus of pseudarthrosis was exposed, removing the fibrous tissue between the bone frag- ments, and the bone stumps were reduced under fluoroscopic control with a C-arm. A bone graft splint was applied as a medial buttress, and internal fixation was performed with a plate and screw at the lateral side. The anatomical planes were then sutured before applying a sterile dressing (Figure 1). (A) (B) (C) Figure 1. 46-year-old patient with a closed 32-B2 fracture. (A) Anteroposterior X-ray showing PSA at the middle third of femoral diaphysis, initially treated with an intramedullary nail, which appear broken at the fracture site. (B) Coronal CT section of the femur showing the PSA focus, the broken nail and posterior bone loss. (C) Anteroposterior X-ray of the femur after revision surgery with lat- eral plate and screws and medial bone graft splint. Figure 1. 46-year-old patient with a closed 32-B2 fracture. (A) Anteroposterior X-ray showing PSA at the middle third of femoral diaphysis, initially treated with an intramedullary nail, which appear broken at the fracture site. (B) Coronal CT section of the femur showing the PSA focus, the broken nail and posterior bone loss. (C) Anteroposterior X-ray of the femur after revision surgery with lateral plate and screws and medial bone graft splint. (B) (C) (B) (A) (C) (B) Figure 1. 46-year-old patient with a closed 32-B2 fracture. 3. Results The sample consisted of ten men and six women, with an average age of 50 years (range 12–82) at the time of trauma, which has been initially treated with an intramedullary nail or plate and screws. Nine cases involved the left side and seven the right side. In all cases, atrophic PSA resulted after a fracture caused by a high-energy trauma, mainly a car or motorcycle event. In our series, eight PSA occurred at the proximal third of the femoral shaft, four at the middle third and four at the distal third. Two patients were obese (body mass index ≥30), four were polytrauma patients with multiple district involvement, one patient had type 1 neuroﬁbromatosis, four patients had multiple comorbidities (diabetes, COPD, and arterial hypertension), three patients were smokers, while none had alcohol abuse issues. In one case, the femoral fracture was linked to a point exposure, classiﬁed as type 1 according to the Gustilo-Anderson classiﬁcation. Eight patients were treated by intramedullary nail, and eight patients by plate and screws. Two patients had undergone an intermediate surgery to dynamise the medullary nail, previously implanted, in order to address the consolidation delay. J. Clin. Med. 2022, 11, 7407 4 of 10 te and All these procedures were performed in other hospitals and came to our observation after ﬁnding the non-union of the fracture. nail, previously implanted, in order to address the consolidation delay. All these procedures were performed in other hospitals and came to our observation after finding the non-union of the fracture. g All patients considered underwent revision surgery with new internal ﬁxation. g All patients considered underwent revision surgery with new internal fixation. The time between the ﬁrst osteosynthesis and the revision surgery for PSA was, on average, 454 days (range, 302–694 days). p g y The time between the first osteosynthesis and the revision surgery for PSA was, on average, 454 days (range, 302–694 days). Seven patients experienced the rupture of the implants previously implanted, while in the remaining nine, the implants were still intact when removed by revision surgery. All patients were treated by the same surgeon and the mean intervention time was 163 min (range 40–236). g y ( g y ) Seven patients experienced the rupture of the implants previously implanted, while in the remaining nine, the implants were still intact when removed by revision surgery. 3. Results The presence of thin shoots of loose fibrous tissue in the interfragmentary space was associated with the loss of bone substance of the fracture stumps, which also appeared sclerotic. The mean follow-up was of 15.4 months (range 6–38). The demographic characteristics of the pa- tients considered and the different treatments applied in our study are summarised in Table 1. (A) (B) Figure 2. 80-year-old patient with 33-A2 femur fracture. (A) Anteroposterior X-ray of the femur showing a PSA is present in the distal third of the femoral diaphysis, associated with mono-com- partmental knee prothesis, broken plate, multiple Kirshner wires and metal cerclages. (B) Antero- posterior knee X-ray showing knee mega prosthesis implantation as revision surgery. Figure 2. 80-year-old patient with 33-A2 femur fracture. (A) Anteroposterior X-ray of the femur show- ing a PSA is present in the distal third of the femoral diaphysis, associated with mono-compartmental knee prothesis, broken plate, multiple Kirshner wires and metal cerclages. (B) Anteroposterior knee X-ray showing knee mega prosthesis implantation as revision surgery. (A) (B) Figure 2. 80-year-old patient with 33-A2 femur fracture. (A) Anteroposterior X-ray of the femur showing a PSA is present in the distal third of the femoral diaphysis, associated with mono-com- partmental knee prothesis, broken plate, multiple Kirshner wires and metal cerclages. (B) Antero- posterior knee X-ray showing knee mega prosthesis implantation as revision surgery. Figure 2. 80-year-old patient with 33-A2 femur fracture. (A) Anteroposterior X-ray of the femur show- ing a PSA is present in the distal third of the femoral diaphysis, associated with mono-compartmental knee prothesis, broken plate, multiple Kirshner wires and metal cerclages. (B) Anteroposterior knee X-ray showing knee mega prosthesis implantation as revision surgery. In all cases, during the revision surgery, a lower level of callus formation at the PSA site was found intraoperatively, compatible with atrophic pseudarthrosis, and probably due to excessive rigidity with a lack of stimulus for the callus formation. The presence of thin shoots of loose ﬁbrous tissue in the interfragmentary space was associated with the loss of bone substance of the fracture stumps, which also appeared sclerotic. The mean follow-up was of 15.4 months (range 6–38). The demographic characteristics of the patients considered and the different treatments applied in our study are summarised in Table 1. Recovery from pseudarthrosis was reported in 13 patients (81% of cases), including patients who had had total hip arthroplasty and mega prosthesis of the knee implants. 3. Results (A) Anteroposterior X-ray showing PSA at the middle third of femoral diaphysis, initially treated with an intramedullary nail, which appear broken at the fracture site. (B) Coronal CT section of the femur showing the PSA focus, the broken nail and posterior bone loss. (C) Anteroposterior X-ray of the femur after revision surgery with lat- eral plate and screws and medial bone graft splint. Figure 1. 46-year-old patient with a closed 32-B2 fracture. (A) Anteroposterior X-ray showing PSA at the middle third of femoral diaphysis, initially treated with an intramedullary nail, which appear broken at the fracture site. (B) Coronal CT section of the femur showing the PSA focus, the broken nail and posterior bone loss. (C) Anteroposterior X-ray of the femur after revision surgery with lateral plate and screws and medial bone graft splint. In three patients, the same surgical approach was implemented, but instead of a bone graft splint, an autologous cortico-cancellous bone graft taken from the iliac crest was placed medially at a non-union level, along with lateral plate and screws ﬁxation. In three patients, the same surgical approach was implemented, but instead of a bone graft splint, an autologous cortico-cancellous bone graft taken from the iliac crest was placed medially at a non-union level, along with lateral plate and screws ﬁxation. Two patients were treated by revisioning the intramedullary nail using an antegrade nail with a larger diameter (Gamma® 3 model, Stryker, Kalamazoo, MI, USA) after further reaming of the medullary canal. g y In two patients with PSA of the proximal third of the femoral diaphysis, an antero- lateral approach to the hip was performed, with a total hip replacement (Accolade® II J. Clin. Med. 2022, 11, 7407 5 of 10 , Ham 5 of 10 , Ham model, Stryker, Kalamazoo, MI, USA). In two patients, the onset of non-union in the dis- tal third of the femoral diaphysis was associated with signiﬁcant bone resorption and marked arthritic degeneration of the knee cartilage. In these cases, it was not possible to proceed with internal ﬁxation, but it was necessary to implant a knee mega prosthesis (LINK® MEGASYSTEM-C® Tumor and Revision System, Waldemar Link GmbH & Co. KG, Hamburg, Germany) (Figure 2). due to excessive rigidity with a lack of stimulus for the callus formation. 3. Results p p p y g p p In three patients, the normal reparative healing processes did not occur in the follow- up period. These patients had multiple comorbidities, such as type 2 diabetes mellitus, hypertension, and COPD. No patient had a history of infection at the fracture site, so all the PSA were considered non-septic. There were no postoperative infections after revision surgery in any patient, although some complications did occur. In the postoperative course, four patients were transfused with concentrated red blood cell units for post-surgical anaemia. A supra-fascial ﬂuid collection with no signs of infection was observed in one J. Clin. Med. 2022, 11, 7407 6 of 10 patient, while another one was left with a dystrophic scar, adhering to the underlying anatomical planes. patient, while another one was left with a dystrophic scar, adhering to the underlying anatomical planes. Table 1. Patient demographics and applied treatments. Table 1. Patient demographics and applied treatments. Patients with Femoral PSA (n = 16) Mean Age (years) 50 (12; 82) Side Sex Left 9 Right 7 Male 10 Open fractures (n) 1 Female 6 Fracture localization (n) High-energy trauma (n) 16 Proximal third 8 First implant rupture (n) 7 Middle third 4 Initial treatment (n) Distal third 4 Intramedullary nail 8 PSA treatment (n) Plate and screw 8 Plate + homologous bone graft 7 Smokers (n) 3 Plate + cortico-cancellous bone graft 3 Plate + screw 8 Intramedullary nail 2 Follow-up (months) 15.4 (range 6–38) Prosthetic implantation 4 Table 1. Patient demographics and applied treatments. Magnetotherapy was used by ﬁve patients to stimulate the bone healing of the PSA focus, leading to a progression of osteo-repairing phenomena. Recovery from pseudarthrosis occurred in all ﬁve patients (100%) initially treated with an intramedullary nail and subsequently treated with plate and screws and bone graft implant. g p Patients who achieved PSA consolidation after neo-osteosynthesis experienced mild thigh and knee pain, muscle hypotonia of varying degrees, reduced knee range of motion (ROM) with ﬂexion block between 90–110◦, and varying degrees of disability. The level of disability was grade 1 in seven cases and grade 2 in two cases. One patient treated with knee mega prosthesis presented decreased joint ROM, with a deﬁcit of 15◦both in ﬂexion and in extension compared with the contralateral knee, and chronic pain, which increased with load and active mobilisation. 3. Results The other patients treated with knee or hip resection prosthesis had a normal postoperative course, with normal or moderately reduced ROM, occasional pain, mild hypotonia, and 1–2 degree of disability. During the follow-up, none of the patients showed intolerance to the ﬁxation implants or their mobilisation and in no case was the implanted material removed. 4. Discussion Earlier studies have presented results comparable with our own. To treat femoral shaft PSA, we often use internal ﬁxation with plate and screws associated with the medial bone graft. This surgery allows for the removal of all the ﬁbrotic tissue interposed. It enables the establishment of a stable, but at the same time ﬂexible, system for adequate bone tissue formation. The use of autologous and homologous bone grafts constitutes a potentially invaluable tool; it makes it possible to rely on growth factors stimulating the formation of the reparative callus, thus obtaining a further stabilisation of the PSA focus, considering that patients with such a condition usually have poor bone quality. However, surgery must always be carefully planned based on fracture type, patient age, the site of the fracture and bone conditions in order to obtain good results both in terms of the healing process and quality of life. The examination of the proposed cases and the scientiﬁc literature allows us to exhibit some orthopaedic-traumatic and medico-legal remarks. The fracture healing process is inﬂuenced by many potentially interfering biological factors. For example, factors related to bone biology and genetic predisposition, patient- related factors such as lifestyle habits and comorbidities and some related to fracture characteristics such as topography or soft tissue injuries [29]. Patient age is very important in determining the recovery process: in children and young adults, the periosteum is rich in osteoblasts and is well vascularised, whereas elderly patients usually exhibit a partially ﬁbrous periosteum, associated with slower callus formation [30]. In patients with osteoporosis, delayed expression of estrogen receptors was detected during the healing process and is associated with reduced callus formation. Nikolaou et al. reported the average consolidation time of femoral shaft fractures treated with a medullary nail in patients with osteoporosis to be three weeks longer than in healthy patients [31]. Furthermore, fracture surgical ﬁxation procedures in patients with osteoporosis are known to incur higher rates of complications, such as loss of reduction, failure implantation, delayed union, or non-consolidation. The activity of osteoprogenitor cells is also inﬂuenced by genetic factors. In fact, the recruitment, differentiation, and proliferation of osteogenic cells during the initial phase of fracture healing are reduced due to the low production of growth factors, such as BMPs and the qualitative and quantitative deﬁcit of MSCs [32,33]. 4. Discussion Pseudarthrosis is a possible complication of femoral shaft fractures that require ad- equate therapeutic planning, which needs to be aimed towards two fundamental goals: mechanical stability to allow the early recovery of joint function and walking, and protec- tion of vascularity, to minimise trophic damages, secondary to excessive cruentation at the PSA focus. Deﬁnitive surgery must ensure adequate stability of the fracture site, as well as accurate bone alignment, to allow for optimal functionality of the operated limb. g p y p The ultimate objective is to reduce the risk of permanent disability, often severe in a patient affected by PSA, especially in young patients incurring such a complication after a high-energy trauma. About possible treatments for PSA, Kouzelis et al. have demonstrated that reaming the femoral canal has an important osteo-inductive biological effect due to reaming debris, which acts as an autologous endogenous graft [22]. Another possible treatment is an internal ﬁxation with plate and screws, a more invasive surgery which allows excision of the ﬁbrous tissue at the PSA focus, direct visualisation of the fracture site, and correction of rotation and mechanical axis [23]. Internal ﬁxation of the plate and screw, however, may not be enough if associated with osteopenia, comminute fracture, or loss of bone stock. High complication rates have been reported in such cases, including misalignment, reduced mobility and persistent J. Clin. Med. 2022, 11, 7407 7 of 10 7 of 10 non-union [24]. For this reason, the use of bone grafts is increasingly associated with internal ﬁxation with a plate to give more stability to the bone, stimulate the normal healing processes and offset bone loss. Chapman and Finkemeier proved the healing in eighteen patients suffering from supracondylar pseudarthrosis of the femur after ﬁxation with plate and screws and bone graft [25]. Wang and Weng treated 13 patients with removal of previously implanted ﬁxation media, internal ﬁxation, and autograft, reaching a 100% union rate [26]. Maimaitiyiming et al. used dual plate and bone grafting in 14 patients with PSA after femoral shaft fractures, achieving healing in all operated patients [27]. g g p p In other cases, as reported by Raju Vaishya et al., the unsuccessful internal ﬁxation of distal femoral shaft fractures, associated with bone loss and arthritis of the knee with or without ligament instability, can be treated with a cemented mega prosthesis, especially in elderly patients [28]. 4. Discussion Manigrasso and O’Connor, in an ex- perimental study on rats, have identiﬁed several genes, such as C57BL/6, DBA/2 and C3H9, capable of inﬂuencing the fracture consolidation process. In mice with C57BL/6 genes, the consolidation of the fracture was achieved in a shorter time [34]. Hofmann et al. have shown reduced cell viability and downregulated gene expression of signalling molecules (IGF-, TGF-β- and FGF) in osteoblasts of patients with hypertrophic bone non-union [35]. It has been reported that polymorphisms and genetic proﬁles of speciﬁc molecules affect both cell differentiation and the generation process of endochondral callus [36]. Such factors may indeed take on increasingly great relevance as personalized medicine based on the unique genomic traits of each individual patient makes further progress [37]. J. Clin. Med. 2022, 11, 7407 8 of 10 8 of 10 In addition, there is no discounting the role plaid by lifestyle habits such as smoking, alcohol and non-steroidal anti-inﬂammatory drugs (NSAIDs) abuse, stress, and poor eating habits, which constitute noteworthy risk factors for non-consolidation [38,39]. In addition, diabetes can impair the healing process of the fracture. Patients receiving adequate insulin therapy have a lower risk of delayed and non-consolidation [40,41]. Several risk factors are closely linked to the type of fracture, such as the mechanism of injury as high or low energy trauma, vascular damage, and impairment of soft tissues. The number, the comminution of the fragments and the extent of the interfragmentary gap affect the healing process [42]. g p In light of such ﬁndings, all factors related to the patient and the fracture’s type need to be thoroughly accounted for during the forensic evaluation of unsuccessful femoral shaft fracture treatment. Such factors often play a decisive role in the development of pseudarthrosis and can determine the negative outcomes of orthopaedic surgical treatments. At the same time, it will also be necessary to verify whether adequate biological therapies have been administered to stimulate the reparative process. Therefore, attention in the evaluation of any proﬁles of medical professional responsibility must be focused on the patients themselves, without being limited to an exclusive judgment on mechanical stability and correctness of internal ﬁxation. The latter is necessary but not enough for a positive outcome of the treatment. We must also always consider the biological environment of the fracture. 4. Discussion From 2020, moreover, the SarsCoV-2 pandemic has been disrupting the regular provision of clinical-instrumental controls, creating management difﬁculties not attributable to the individual surgeons, limited in part by the introduction of the applications of telemedicine which made it possible to overcome some organisational difﬁculties [42]. Conﬂicts of Interest: The authors declare no conﬂict of interest. Conﬂicts of Interest: The authors declare no conﬂict of interest. 5. Conclusions The literature herein analysed and the results obtained from the analysis of our sample of patients underline the importance of the anatomo-functional damage caused by a femoral shaft pseudarthrosis, which can determine a 3 or 4 disability level, with severe or total functional impotence on the injured lower limb. For the treatment of femoral pseudarthrosis, an accurate assessment of the patient’s general condition, age, type of fracture and bone quality is essential to choosing the most suitable treatment to create the best possible conditions conducive to bone healing. From a medico-legal perspective, it is apparent that chronic consolidation outcomes after revision surgery of femoral shaft PSA cannot be compared with the results after a consolidated femoral fracture with mild functional repercussions. The PSA of the femur recovered after a second osteosynthesis with bone grafts is burdened by a major anatomical and functional impairment. Such impairment is caused by the double surgical access, the second one wider and more complex; by a morphologically and biomechanically less valid callus than the ﬁrst intention consolidation one; by mild and unavoidable angular or rota- tional bone deformities; by greater residual hypotonia; by more signiﬁcant repercussions in joint functions linked to the long therapeutic path and the myotendinous scars, especially of the knee. The physical damage resulting from the outcomes of femoral shaft PSA consolidated after a revision osteosynthesis treatment and the bone grafts, will be more substantial than the permanent sequelae following a femoral diaphyseal fracture consolidated ab initio. The presence of any limitations at the hip and/or knee range of motion, deformity and/or dysmetria will require an adequate medico-legal evaluation. J. Clin. Med. 2022, 11, 7407 9 of 10 Author Contributions: Conceptualization, G.B., S.F., L.B.P., M.G., R.A. and S.Z.; methodology, G.B., S.F. and S.Z.; validation, G.B., S.F., Q.A.P., R.A., L.B.P. and S.Z.; investigation, G.B., S.F., L.B.P. and R.A.; data curation, G.B., S.F. and R.A.; writing—original draft preparation, S.F., G.B., L.B.P. and S.Z.; writing—review and editing, G.B., S.F. and R.A.; visualisation, G.B., S.F., Q.A.P., R.A. and S.Z.; supervision, G.B., S.F., Q.A.P., M.G. and S.Z. All authors have read and agreed to the published version of the manuscript. Funding: This research received no external funding. Funding: This research received no external funding. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable. Informed Consent Statement: Not applicable. 5. Conclusions Data Availability Statement: The data presented in this case report are available on request from the corresponding author. References 1. 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https://openalex.org/W4385388751	http://jtam.ulm.ac.id/index.php/mpj/article/download/1945/1019	Indonesian	null	EVALUASI PEMBELAJARAN PJOK PADA SAAT PANDEMI COVID-19 DI SEKOLAH MENENGAH PERTAMA NEGERI DI KECAMATAN BANJARMASIN TIMUR	Stabilitas	2,023	cc-by	2,387	Abstrak Penelitian yang penulis lakukan disini adalah menggunakan penelitian kuantitatif dalam metode ex post facto. Penelitian ini dilakukan di Sekolah Menengah Pertama Kecamatan Banjarmasin Timur dan informan dalam penelitian ini adalah guru Pendidikan Jasmani Olahraga dan Kesehatan. Penulis menggunakan pembagian kuesioner atau angket dan dokumen autentik dari guru yang digunakan saat penilaian. Di dalam pembahasannya digunakan deskriptif kuantitatif yaitu menemukan pengetahuan yang menggunakan data beruapa angka sebagai alat menganalisis. Dari hasil penelitian yang dilakukan penilaian secara daring pada mata pelajaran PJOK di Sekolah Menengah Pertama Negeri Kecamatan Banjarmasin Timur sudah cukup baik, namun belum sepenuhnya memenuhi kaidah penilaian sesuai buku panduan penilaian revisi 2018. Kata kunci: evaluasi pembelajaran PJOK. Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 EVALUASI PEMBELAJARAN PJOK PADA SAAT PANDEMI COVID-19 DI SEKOLAH MENENGAH PERTAMA NEGERI DI KECAMATAN BANJARMASIN TIMUR Masri, H. M. Mulhim dan Abd. Hamid Penddikan Jasmani JPOK FKIP Universitas Lambung Mangkurat, Banjarbaru masri.masri0309@gmail.com Keywords: PJOK Learning Evaluation. METODE Kuantitatif. pengumpulan data memakai teknik dokumentasi dan observasi terstruktur. Penelitian ini menggunakan jenis kuantitatif dengan metode pra-eksperimental. Instrumen Penelitian Metode dengan skala likert, Menurut Arikunto (2012) dalam Jannah et al (2017), “Hasil perhitungan ditafsirkan dengan menggunakan tafsiran persentase. Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 Subjek Penelitian Populasi dari penelitian ini guru SMP Negeri di Kecamatan Banjarmasin Timur. Sampel penelitian berjumlah 10 guru PJOK. Teknik pengambilan sampel merupakan Total Sampling (sampel jenuh). PENDAHULUAN Dalam (UU Nomor 20 Tahun 2003) Tujuan pendidikan nasional adalah untuk mengembangkan kemampuan dan membentuk watak serta peradaban bangsa yang bermartabat dalam rangka mencerdaskan kehidupan bangsa. Dengan tujuan pendidikan nasional. Tujuan PJOK bisa dapat senada dengan tujuan pendidikan nasional. Media pembelajaran Penjas yang dibuat berupa video dan youtube yang digunakan untuk menambah variasi pembelajaran daring agar tetap menarik bagi peserta didik. Pada media yang dibuat tersebut berupa materi-materi gerakan Penjas yang dipraktikkan Guru Penjas sendiri dibuat video dan youtube kemudian dishare melalui whatsapp ke peserta didik saat guru mengajar. menyikapi kondisi pandemi covid-19 yang belum kunjung berakhir dan sistem pembelajaran masih secara daring guru harus kreatif dalam mensiasati pembelajaran daring dengan memaksimalkan media. Evaluasi adalah suatu pemahaman atau penilaian pertumbuhan dan perkembangan peserta didik yang ditujukan pada nilai-nilai yang ditetapkan dalam kurikulum, oleh dengan itu tujuan penilaian adalah untuk memperoleh bahan atau informasi sebagai pembuktian tentang kemampuan atau keberhasilan peserta didik. Evaluasi pembelajaran bertujuan untuk menentukan kemajuan atau hasil belajar siswa yang selamjutnya sebagai laporan kepada orang tua peserta didik atau wali kelas, jaminan kenaikan kelas dan penentuan kelulusan peserta didik. Selain itu evaluasi pembelajaran berencana untuk menempatan peserta didik ke dalam situasi pendidikan dan pembelajaran yang tepat sesuai dengan tingkat kemampuan, minat dan berbagai karakteristik yang dimiliki. Pandemi covid-19 berdampak pada dunia pendidikan. Masalah yang muncul dalam pembelajaran PJOK disekolah di antaranya pembelajaran kurang sesuai RPP, dan orang tua kesulitan mendampingi kegiatan belajar anak, serta keterbatasan akses internet dan kemampuan aplikasi pada fitur-fitur online. Sehingga diperlukan evaluasi agar bisa terlaksana dengan baik. Dalam hal ini maka peneliti sangat tertarik untuk mengetahui evaluasi pembelajaran PJOK pada saat pandemi covid-19 di Sekolah Menengah Pertama di Kecamatan Banjarmasin Timur. Abstract The author is conducting this study utilizing quantitative research and the ex post facto method. Teachers of Physical Education, Sports, and Health were the informants in this study, which took place at a Junior High School in East Banjarmasin District. During the assessment, the author employs the distribution of questionnaires or questionnaires and authentic papers from teachers. Quantitative descriptive analysis is used in the discussion to find information that employs data in the form of numbers as an analytical tool. According to the findings of a study done at the State Junior High Schools of Banjarmasin Timur District, online evaluations of PJOK subjects are fairly good, but they do not fully meet the assessment standards according to the 2018 revised assessment guidebook. Keywords: PJOK Learning Evaluation. 140 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin HASIL PENELITIAN 1.Faktor Kognitif Dalam Evaluasi Secara Daring. No Norma Kategori Frekuensi Persentasi 1. ≥ 12 Sangat Baik 0 0% 141 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga Masri, H. M. Mulhim, Abd. Hamid JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Volume 4 No 2, Juni 2023 2.Faktor Afektif dalam Evaluasi Secara Daring. 3.Faktor Psikomotorik dalam Evaluasi Secara Daring. 2. 10-11 Baik 5 50% 3. 9-9 Cukup 3 30% 4. 8-8 Kurang 2 20% 5. ≤7 Sangat Kurang 0 0% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 13 Sangat Baik 1 10% 2. 10-12 Baik 3 30% 3. 8-9 Cukup 3 30% 4. 6-7 Kurang 3 30% 5. ≤5 Sangat Kurang 0 0% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 14 Sangat Baik 1 10% 2. 11-13 Baik 2 20% 3. 8-10 Cukup 5 50% 4. 5-7 Kurang 2 20% 5. ≤4 Sangat Kurang 0 0% Total 10 100% Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin 2.Faktor Afektif dalam Evaluasi Secara Daring. 3.Faktor Psikomotorik dalam Evaluasi Secara Daring. 2. 10-11 Baik 5 50% 3. 9-9 Cukup 3 30% 4. 8-8 Kurang 2 20% 5. ≤7 Sangat Kurang 0 0% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 13 Sangat Baik 1 10% 2. 10-12 Baik 3 30% 3. 8-9 Cukup 3 30% 4. 6-7 Kurang 3 30% 5. ≤5 Sangat Kurang 0 0% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 14 Sangat Baik 1 10% 2. 11-13 Baik 2 20% 3. 8-10 Cukup 5 50% 4. 5-7 Kurang 2 20% 5. ≤4 Sangat Kurang 0 0% Total 10 100% 2 F kt Af ktif d l E l i S D i 2. 10-11 Baik 5 50% 3. 9-9 Cukup 3 30% 4. 8-8 Kurang 2 20% 5. ≤7 Sangat Kurang 0 0% Total 10 100% 2.Faktor Afektif dalam Evaluasi Secara Daring. 2.Faktor Afektif dalam Evaluasi Secara Daring. No Norma Kategori Frekuensi Persentasi 1. ≥ 13 Sangat Baik 1 10% 2. 10-12 Baik 3 30% 3. 8-9 Cukup 3 30% 4. 6-7 Kurang 3 30% 5. ≤5 Sangat Kurang 0 0% Total 10 100% 3.Faktor Psikomotorik dalam Evaluasi Secara Daring. No Norma Kategori Frekuensi Persentasi 1. ≥ 14 Sangat Baik 1 10% 2. 11-13 Baik 2 20% 3. 8-10 Cukup 5 50% 4. HASIL PENELITIAN 5-7 Kurang 2 20% 5. ≤4 Sangat Kurang 0 0% Total 10 100% 142 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga Masri, H. M. Mulhim, Abd. Hamid JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Volume 4 No 2, Juni 2023 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin 4. Faktor Proses Evaluasi Secara Daring. 5.Faktor Kegiatan Pembelajaran Secara Daring. 6.Faktor Teknik Penilaian Secara Daring. PEMBAHASAN No Nor ma Kategori Frekuen si Persentasi 1. ≥ 36 Sangat Baik 0 0% 2. 31-35 Baik 4 40% 3. 27-30 Cukup 4 40% 4. 23-26 Kurang 1 10% 5. ≤22 Sangat Kurang 1 10% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 20 Sangat Baik 0 0% 2. 16-19 Baik 4 40% 3. 13-15 Cukup 4 40% 4. 9-12 Kurang 2 20% 5. ≤8 Sangat Kurang 0 0% Total 10 100% No Norma Kategori Frekuensi Persentasi 1. ≥ 26 Sangat Baik 0 0% 2. 22-25 Baik 3 30% 3. 18-21 Cukup 5 50% 4. 14-17 Kurang 1 10% 5. ≤13 Sangat Kurang 1 10% Total 10 100% 4. Faktor Proses Evaluasi Secara Daring. 5.Faktor Kegiatan Pembelajaran Secara Daring. No Nor ma Kategori Frekuen si Persentasi 1. ≥ 36 Sangat Baik 0 0% 2. 31-35 Baik 4 40% 3. 27-30 Cukup 4 40% 4. 23-26 Kurang 1 10% 5. ≤22 Sangat Kurang 1 10% Total 10 100% 4. Faktor Proses Evaluasi Secara Daring. 5.Faktor Kegiatan Pembelajaran Secara Daring. No Norma Kategori Frekuensi Persentasi 1. ≥ 20 Sangat Baik 0 0% 2. 16-19 Baik 4 40% 3. 13-15 Cukup 4 40% 4. 9-12 Kurang 2 20% 5. ≤8 Sangat Kurang 0 0% Total 10 100% 6.Faktor Teknik Penilaian Secara Daring. No Norma Kategori Frekuensi Persentasi 1. ≥ 26 Sangat Baik 0 0% 2. 22-25 Baik 3 30% 3. 18-21 Cukup 5 50% 4. 14-17 Kurang 1 10% 5. ≤13 Sangat Kurang 1 10% Total 10 100% PEMBAHASAN Berdasarkan kuesioner pelaksanaan pembelajaran daring ini baik rencana Pelaksanaan Pembelajaran (RPP), Media atau pelaksanaan yang dijalankan masuk kategori baik. Dilihat dari persentase hasil penelitian sebesar 40% dan termasuk dalam kategori baik. Berdsarakan dari dokumen yang di terima sesuai. Dimana, ditinjau dari perencanaan sekolah sudah menggunakan Rpp sesuai 143 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 pembelajaran daring, materi yang diajarkan sesuai dengan KI dan KD yang di pilih, dan materi pembelajaran sesuai dengan Rpp. Tjandra, D. S. (2020), mengatakan “bahwa guru hanya memfasilitasi dengan perpustakaan kelas, modul, buku teks, serta buku-buku pendukung, dan yang terpenting akses internet, serta menyediakan beberapa komputer untuk para siswa yang tidak membawa laptop.”. pembelajaran daring, materi yang diajarkan sesuai dengan KI dan KD yang di pilih, dan materi pembelajaran sesuai dengan Rpp. Tjandra, D. S. (2020), mengatakan “bahwa guru hanya memfasilitasi dengan perpustakaan kelas, modul, buku teks, serta buku-buku pendukung, dan yang terpenting akses internet, serta menyediakan beberapa komputer untuk para siswa yang tidak membawa laptop.”. Dari hasil perhitungan statistik berdasarkan kuesioner, menunjukkan bahwa instrumen yang digunakan pada ranah afektik, kognitif, dan psikomotorik memiliki persentasi rata-rata 30% yang berarti tidak mengalami kekurangan. Pada ranah afektif menunjukkan persentase 30% instrumen yang dikategorikan baik. Pada ranah Kognitif, penelitian menunjukkan persentase dengan angka 50% pada kategori baik. Pada ranah psikomotorik, penelitian menunjukkan persentase dengan 50% yaitu pada kategori cukup baik. Dapat diartikan bahwa instrumen penilaian secara daring yang digunakan guru Sekolah Menengah Pertama Negeri Di Kecamatan Banjarmasin Timur pada ranah afektif adalah baik, pada ranah kognitif adalah baik, dan pada ranah psikomotorik adalah cukup baik. Hal ini sejalan dengan buku Panduan Penilaian K-13 revisi 2018 yang menganjurkan pelaksanaan Penilaian Harian (PH) agar dapat melaksanakan dengan penilaian tiga ranah sekaligus yaitu ranah afektif, kognitif, dan psikomotorik. Pada teknik penilaian ini baik pada ranah afektif, kognitif, maupun psikomotorik dari hasil penelitian menunjukkan bahwa teknik penilaian yang digunakan adalah baik. Hal ini dibuktikan melalui hitungan statistik dari kuesioner dengan persentase sebesar 50% pada kategori cukup. Dapat diartikan bahwa teknik penilaian secara daring yang digunakan guru di Sekolah Menengah Pertama Negeri Di Kecamatan Banjarmasin Timur adalah cukup baik. Setiadi (2016) menyebutkan bahwa “Penggunaan teknik penilaian disesuaikan dengan kebutuhan-kebutuhan yang dapat menunjang program pengajaran seperti kompetensi dasar yang akan dicapai.”. PEMBAHASAN Pada proses penilaian yang dilakukan guru secara daring di SMPN di Kecamatan Banjarmasin Timur berjalan cukup baik. Hal ini dibuktikan melalui hitungan statistik berdasarkan kuesioner dengan persentase sebesar 40% yaitu termasuk dalam kategori dengan proses penilaian yang cukup baik. Berdasarkan dokumen yang diterima peneliti pada proses penilaian pada prosesnya yaitu di luar jam pelajaran karena guru menilai setelah peserta didik mengumpulkan tugasnya dan jangka waktu mengerjakan cukup lama. Untuk pelaksanaan penilaian ada sebagian guru melaksanakan penilaian pada ranah kognitif dan psikomotorik setiap kali pertemuan. SARAN Guru PJOK hendaknya menyesuaikan dan diharapkan membuat laporan hasil penilaian secara daring baik pada ranah afektif, kognitif, dan psikomotorik dalam penilaian harian (PH) dengan lebih baik lagi, karena penelitian ini sebatas mengetahui instrumen, teknik, dan proses penilaian secara daring di SMP. KESIMPULAN Penilaian secara daring pada mata pelajaran PJOK di Sekolah Menengah Pertama Negeri Kecamatam Banjarmasin Timur sudah cukup baik, namun belum sepenuhnya memenuhi kaidah penilaian sesuai buku panduan penilaian revisi 2018. Berdasarkan faktor perencanaan dan kegiatan pembelajaran yang dilaksanakan di SMP Negeri Kecamatan Banjarmasin Timur adalah baik, hal ini dibuktikan dengan kesesuaian data kuesioner dengan dokumen. Faktor instrumen yang digunakan berdasarkan data autentik instrumen penilaian pada ranah afektif tidak ada, pada ranah kognitif melalui penugasan dan tes tertulis, dan pada ranah psikomotor berupa video unjuk kerja dan produk. Pada faktor teknik penilaian berdasarkan data autentik teknik penilaian hanya terlaksana pada ranah kognitif dan psikomotor. Faktor proses penilaian berdasarkan data autentik, proses penilaian tersebut hanya terlaksana pada ranah kognitif dan psikomotorik. Pada pelaksanaannya tidak semua sekolah melaksanakan disetiap pertemuan dan penilaian hanya dilakukan dari guru ke murid di luar kegiatana PBM. 144 Stabilitas :Jurnal Pendidikan Jasmani dan Olahraga JPOK FKIP Universitas Lambung Mangkurat Banjarmasin Masri, H. M. Mulhim, Abd. Hamid Volume 4 No 2, Juni 2023 Pjok, K., Viii, K., & Aji, B. S. (2016). Pengembangan Instrumen Penilaian Pengetahuan Mata Pelajaran Pendidikan Jasmani Olahraga dan. April 2015, 267–283. DAFTAR PUSTAKA Asrul, Ananda, R., & Rosinta. (2014). Evaluasi Pembajalaran. In Ciptapustaka Media. Dewi, W. A. F. (2020). Dampak COVID-Dewi, W. A. F. (2020). Dampak COVID-19Dewi, W. A. F. (2020). Dampak COVID-Dewi, W. A. F. (2020). Dampak COVID-19 terhadap Implementasi Pembelajaran Daring di Sekolah Dasar. Edukatif : Jurnal Ilmu Pendidikan, 2(1), 55–61. https://doi.org/10.31. Edukatif : Jurnal Ilmu Pendidikan, 2(1), 55–61. Herlina, H., & Suherman, M. (2020). Potensi Pembelajaran Pendidikan Jasmani Olahraga Dan Kesehatan (Pjok) Di Tengah Pandemi Corona Virus Disease (Covid)-19 Di Sekolah Dasar. Tadulako Journal Sport Sciences And Physical Education, 8(1), 1–7. http://jurnal.untad.ac.id/jurnal/index.php/PJKR/article/view/16186 Junaedi, A., & Wisnu, H. (2016). Survei Tingkat Kemajuan Pendidikan Jasmani, Olahraga, Dan Kesehatan Di Sma, Smk, Dan Ma Negeri Se-Kabupaten Gresik. Jurnal Pendidikan Olahraga Dan Kesehatan, 3(3), 834–842. Lanziotti, V. S., De Souza, D. C., & Marques, E. T. A. (2020). Coronavirus Disease 2019: Understanding Immunopathogenesis Is the “holy Grail” to Explain Why Children Have Less Severe Acute Disease. Pediatric Critical Care Medicine, 1022–1023. https://doi.org/10.1097/PCC.0000000000002513 Mustafa, P. S., Winarno, M. E., & Supriyadi, S. (2019). Penilaian Pendidikan Jasmani, Olahraga, dan Kesehatan pada Sekolah Menengah Pertama Negeri Kota Malang. Jurnal Pendidikan: Teori, Penelitian, Dan Pengembangan, 4(10), 1364. https://doi.org/10.17977/jptpp.v4i10.12845 Nur, M., Pradipta, D. G., & Maliki, O. (2020). Efektivitas Pembelajaran Daring Mata Pelajaran Pjok Siswa Kelas Ix Di Mtsn 2 Semarang Selama Pandemi Covid–19. Seminar Nasional Keindonesiaan (FPIPSKR), 2(1), 108–125. http://conference.upgris.ac.id/index.php/snk/article/view/1139 Pjok, K., Viii, K., & Aji, B. S. (2016). Pengembangan Instrumen Penilaian Pengetahuan Mata Pelajaran Pendidikan Jasmani Olahraga dan. April 2015, 267–283. 145 145
https://openalex.org/W4251716614	https://www.qeios.com/read/R6V77I/pdf	English	null	Community Health Nurse Specialist	Definitions	2,020	cc-by	73	Qeios · Definition, February 7, 2020 Open Peer Review on Qeios Open Peer Review on Qeios Community Health Nurse Specialist National Cancer Institute Qeios ID: R6V77I · https://doi.org/10.32388/R6V77I Source National Cancer Institute. Community Health Nurse Specialist. NCI Thesaurus. Code C20834. A registered nurse prepared at the master's degree level or higher who specializes in exploring and meeting the needs of select populations in community contexts. (UMD, SON) Qeios ID: R6V77I · https://doi.org/10.32388/R6V77I 1/1
https://openalex.org/W4229524214	https://www.qeios.com/read/ZPVEK2/pdf	English	null	SPN wt Allele	Definitions	2,020	cc-by	80	Qeios · Definition, February 2, 2020 Open Peer Review on Qeios SPN wt Allele National Cancer Institute National Cancer Institute Qeios ID: ZPVEK2 · https://doi.org/10.32388/ZPVEK2 Open Peer Review on Qeios Source National Cancer Institute. SPN wt Allele. NCI Thesaurus. Code C101152. Human SPN wild-type allele is located in the vicinity of 16p11.2 and is approximately 8 kb in length. This allele, which encodes leukosialin protein, plays a role in both chemotaxis and leukocyte migration. Qeios ID: ZPVEK2 · https://doi.org/10.32388/ZPVEK2 1/1
https://openalex.org/W4391639903	https://periodicos.uem.br/ojs/index.php/ActaSciAnimSci/article/download/64312/751375156936	English	null	Diet supplementation formulated with Bacillus sp. SMIA-2 and its enzymes for Nile tilapia: zootechnical performance and effects on intestinal morphometry	Acta Scientiarum. Animal Sciences	2,023	cc-by	6,836	Acta Scientiarum http://periodicos.uem.br/ojs ISSN on-line: 1807-8672 Doi: 10.4025/actascianimsci.v46i1.64312 Acta Scientiarum http://periodicos.uem.br/ojs ISSN on-line: 1807-8672 Doi: 10.4025/actascianimsci.v46i1.64312 AQUACULTURE Diet supplementation formulated with Bacillus sp. SMIA-2 and its enzymes for Nile tilapia: zootechnical performance and effects on intestinal morphometry Priscilla Cortizo Costa Pierro2, Pedro Pierro Mendonça2, Paola de Oliveira Santos2, Thayna de Souza Pardo2, Thaianara Galdino do Nascimento Assis2, Samuel Oliveira da Silva2, David Carvalho dos Santos Ribeiro2 and Meire Lelis Leal Martins1* 1Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense, 28013-602, Campos dos Goytacazes, Rio de Janeiro, Brazil. 2Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, Campus de Alegre, Alegre, Espírito Santo, Brazil. *Author for correspondence. E- mail: meire@uenf.br ABSTRACT. The use of probiotics and exogenous enzymes in fish feed is a promising alternative to improve animal performance. This study evaluated the feasibility of applying Bacillus sp. SMIA-2 and its enzymes as supplements in the diet for juvenile tilapia. The effect of adding different concentrations of Bacillus sp. SMIA-2 and its enzymes in food on zootechnical development, intestinal morphometry of animals was analyzed. The bacteria could be recovered from the intestines of animals, demonstrating its ability to survive gastric and bile acids. The comparative study of SMIA-2 with commercial products showed a significant effect on individual food intake, final weight and weight gain in all treatments. Gut length, villus height and intestinal coefficient were an advantage of SMIA-2 compared to commercial products and the control group. Therefore, the inclusion of Bacillus sp. SMIA-2 and its enzymes in fish feed may represent a viable alternative to improve animal development and significantly increase intestinal villi, contributing to nutrient absorption and animal health. Keywords: aquaculture; Bacillus; enzymes; animal nutrition; Oreochromis niloticus. Received on July 11 2022. Accepted on February 28 2023. Received on July 11 2022. Accepted on February 28 2023. Received on July 11 2022. Accepted on February 28 2023. Material and methods The experiment was conducted in the Laboratory of Nutrition and Production of Ornamental Species (Laboratório de Nutrição e Produção de Espécies Ornamentais – LNPEO) in the Campus of Alegre, state of Espírito Santo, between March and May 2021. The study was approved by the Animal Research Ethics Committee of the Federal Institute of Espírito Santo (Instituto Federal do Espírito Santo), registered under the number 23149.001859/2021-56. The synbiotic product used in the experiments was supplied by the Food Technology Laboratory from the State University of Northern Rio de Janeiro (Universidade Estadual do Norte Fluminense Darcy Ribeiro - UENF). It contained 7.3 Log spores mL-1 of Bacillus sp. SMIA-2 and the following enzymes (U mL-1): 11.46 proteases, 1.10 avicelases (avicel hydrolyzing enzymes – insoluble cellulose), 0.13 carboxymethyl cellulose (carboxymethyl cellulose hydrolyzing enzymes - soluble cellulose), 0.30 xylanases, 0.29 amylases, and 0.23 polygalacturonases. Initially, an experiment was conducted to check the effect of adding different concentrations of Bacillus sp SMIA-2 in the feed on the zootechnical development and intestinal morphometry of tilapia juveniles. The experimental design adopted was the completely randomized design with five concentrations of SMIA-2 (g kg-1 feed): 0.0; 0.383 g; 0.765 g; 1.147 g; 1.531 g, and five replications of 15 juveniles in each treatment, resulting in 25 experimental units. The experimental units (EU) contained 15 juveniles and the aquarium, with a volume of 50 L. After, the zootechnical development of tilapia juveniles using feed supplemented with SMIA-2 and its enzymes was compared to two commercial products tested in the same conditions. A completely randomized design (CRD) with four treatments was adopted: Treatment 1 (T1): control feed; Treatment 2 (T2): feed with spores and enzymes of Bacillus sp. SMIA-2; Treatment 3 (T3): feed with a commercial product containing Bacillus licheniformis and enzymes (0.250 g kg-1); Treatment 4 (T4): feed including a commercial product containing Bacillus coagulans, Bacillus subtilis, Bacillus licheniformis, and enzymes (0.100 g kg-1). The doses of the commercial products followed the instructions of manufacturers. The inclusion dose of Bacillus sp. SMIA-2 was 0.765 g kg-1. Each treatment had six replications, with 15 tilapia juveniles each, resulting in 24 experimental units (EU) with a 50 L volume each. Each EU had water recirculation, individual water input and output, physical, chemical, and biological filtering, an overflow box with a water pump of 3/4 hp, and heaters with thermostats. Page 2 of 11 Pierro et al. This study evaluated the feasibility of using Bacillus sp. SMIA-2 and its enzymes as supplement in diets formulated for tilapia (Oreochromis niloticus). Introduction Tilapia is currently the main fish species farmed in Brazil (Figueiredo et al., 2022). In 2020, 486,155 tons of the species were produced, making the country the fourth largest tilapia producer globally (Peixe BR, 2021). Expansion of tilapia farming requires high amounts of sustainable ingredients for nutritionally balanced fish feed (Furuya, Cruz, & Gatlin III, 2023). Plant-based feed, such as flours, oilseeds, legumes, and cereal by- products, is the main ingredient in formulations. Nevertheless, there are limitations in their inclusion levels due to the presence of antinutritional factors, such as phytin, non‐starch polysaccharides, and protease inhibitors, which may impair the use of nutrients, fish development, and overall health (Castillo & Gatlin, 2015). Adding enzymes in feed formulation is an alternative to remove or reduce antinutritional factors, improving nutrient digestion and, consequently, the zootechnical performance of animals (Xu, Zheng, Dong, Ai, & Mai, 2022). Microbial enzymes, especially those produced by the genus Bacillus, have been widely explored for this aim. Bacillus SMIA-2, an aerobic, thermophilic, and spore-forming bacterium (Souza & Martins, 2001), can produce thermostable enzymes, such as proteases (Silva, Delatorre, & Martins, 2007), amylases (Carvalho, Corrêa, Silva, Viana, & Martins, 2008), pectinases (Andrade, Delatorre, Ladeira, & Martins, 2011), and cellulases (Ladeira, Cruz, Delatorre, Barbosa, & Leal Martins, 2015; Cruz, Moraes, Costa, Barbosa, & Martins, 2019). Bernardo et al. (2020) recently sequenced the SMIA-2 genome and detected no virulence gene. According to the authors, this strain is 100% similar to Bacillus licheniformis Gibson 46 (ATCC 14580T). Due to the spore-forming ability of the genus Bacillus, which resists gastric and biliary acids, it can survive in the gastrointestinal tract and remain in high concentration in the intestine (Lee & Kim, 2011). Therefore, Bacillus species are currently widely studied for the human, animal, and aquaculture nutrition as growth promoters and agents of competitive exclusion (Latorre et al., 2016). Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Bacillus as additives in fish diets Page 3 of 11 Page 3 of 11 After the adaptation period, tilapia juveniles were selected according to their initial biometrics. Afterwards, total length, standard length, and height were measured with a caliper and weighed on a digital scale with four decimal places and three precision numbers. All fish were sedated with 0.25 mL eugenol per liter of water. Fish had, on average, an initial weight of 1.14 g and a mean total length of 4.32 cm. After initial biometrics, fish were evenly distributed in the EUs. For the control and monitoring of water quality in the experimental systems, the following water quality parameters were measured daily: Dissolved oxygen, Higher water temperature, Lower water temperature, and the temperature at the time. Dissolved oxygen and temperature were measured with an oximeter and a digital thermometer. Values for pH and conductivity were measured with a digital pH meter and an electrical conductivity meter, respectively, and ammonium nitrite and nitrate with a digital microprocessor photocolorimeter every two days. All devices were calibrated before each measurement. Zootechnical performance For the final biometrics of the experiments, animals were euthanized following the Guidelines for Euthanasia Practices of the National Council for the Control of Animal Experimentation – CONCEA – for bony and cartilaginous fish, using eugenol at the dose of 5 mL/liter of water. Variables used to quantify the development of juveniles were weight, total length, standard length, height, weight gain, apparent food conversion, specific growth rate, condition factor, protein efficiency ratio, energy efficiency ratio, survival rate, and batch uniformity, as follows: Weight Gain: WG= Wf - Wi (g). Wf = average final weight (g), and Wi = average initial weight (g Weight Gain: WG= Wf - Wi (g). Wf = average final weight (g), and Wi = average initial weight (g). •Feed consumption: FC (g) in the period = (Feed supplied – Feed leftovers) / number of animals. •Feed Conversion ratio: FCR= FC / WG. Weight Gain: WG= Wf - Wi (g). Wf = average final weight (g), and Wi = average initial weight (g). •Feed consumption: FC (g) in the period = (Feed supplied – Feed leftovers) / number of animals. •Feed Conversion ratio: FCR= FC / WG. •Feed consumption: FC (g) in the period = (Feed supplied – Feed leftovers) / number of animals. •Feed consumption: FC (g) in the period = (Feed supplied – Feed leftovers) / number of animals. •Feed Conversion ratio: FCR= FC / WG. •Specific Growth rate: SGR= ((lnWf – lnWi)) /Tta) x 100. •Condition Factor: K= (W x 100) / L 3. W= weight (g), and L= length (cm). •Condition Factor: K= (W x 100) / L 3. W= weight (g), and L= length (cm). •Survival Rate: SR= (Nf / Ni) x100. Nf = final number of fish, and Ni= initial number of fish/experimental unit. •Protein efficiency ratio: PER= (WG/ (FC X %RP of the diet)) X 100. WG= weight gain, and FC = feed consumption. •Survival Rate: SR= (Nf / Ni) x100. Nf = final number of fish, and Ni= initial number of fish/experimental unit. •Protein efficiency ratio: PER= (WG/ (FC X %RP of the diet)) X 100. WG= weight gain, and FC = feed consumption. •Energy efficiency ratio: EER= (WG/ (FC X %RP of the diet)) X 100. WG= weight gain, and FC = feed consumption. •Energy efficiency ratio: EER= (WG/ (FC X %RP of the diet)) X 100. WG= weight gain, and FC = feed consumption. Histopathological analysis For villi analysis, intestinal fragments with around 3 cm length and 5 mm thickness were tak section in two different intestinal segments (duodenum and ileum). Intestinal samples were fixed in neutral buffered formalin at 10%, and stored until processing. Cleavage was conducted, and fragments were placed into histology cassettes for paraffin embedding. The material processing consists of the following steps: sample dehydration in increasing alcohol series (70, 85, 95%, absolute alcohol I, II, and III) during one hour for each sample. After dehydration, samples were cleared in xylol solutions I, II, and III, where they remained for an hour in each concentration. Then, they were immersed in paraffin I and II for an hour in each. Soon after, samples were embedded in paraffin mixed with beeswax, and stored in a freezer at -20°C. Blocks were cross sectioned with a microtome to 4 µm thickness. Slides were stained with hematoxylin and eosin and observed under a light microscope. Villi height was observed and measured with an ocular micrometer (Figure 1). Material and methods The experimental units were siphoned for partial water exchange (PWE) with approximately 20% of the useful olume every three days. Feed was supplied three times a day until satiation. The experimental feeds were formulated for tilapia juveniles, containing 36% crude protein (CP) and 3,100 Kcal digestible energy (DE). Each treatment received the same basal feed but added with Bacillus sp. SMIA-2 in different concentrations. Feed ingredients were ground and mixed in the proportions mentioned above (Table 1). After mixing, ingredients were moistened with water at 55ºC and mixed. Only after this process, the feed was pelleted. Later, ingredients were dried outdoors in the shade and duly stored in a refrigerator or freezer to be used during the experiment. The additive was included on the top of the feed. Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Table 1. Composition percentage and nutritional value of the basal experimental feed used in the fish diet. Ingredients Soybean meal 64.85 % Cornmeal 20.14 % Fish meal 10.00 % Wheat bran 3.00 % Soybean oil 2.00 % Nutritional value Crude protein 36% Digestible energy 3,101.2 Kcal Table 1. Composition percentage and nutritional value of the basal experimental feed used in the fish diet. Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Table 1. Composition percentage and nutritional value of the basal experimental feed used in the fish diet. Ingredients Soybean meal 64.85 % Cornmeal 20.14 % Fish meal 10.00 % Wheat bran 3.00 % Soybean oil 2.00 % Nutritional value Crude protein 36% Digestible energy 3,101.2 Kcal Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Effect of adding different concentrations of Bacillus sp SMIA-2 in the diet on the zootechnical development and intestinal morphometry of tilapia juveniles Mean values of quality parameters measured during the experimental period were temperature – 26.77°C; dissolved oxygen – 3.207 mg L-1; ammonium 0.062 mg L-1, and potential of hydrogen (pH) – 8.055. Water quality monitoring is essential for aquaculture productivity, even when farming more rustic species, such as tilapia. According to Freitas et al. (2022), tilapia presents thermal comfort at temperatures between 27 and 30°C. Higher temperatures (above 38°C) can lead to mortality due to the reduced levels of dissolved oxygen in water. When the temperature is low, there is lower resistance, and the animal is susceptible to illnesses, besides increasing cortisol levels (Falcon, Barros, Pezzato, Solarte, & Guimarães, 2008). Although the dissolved oxygen is below the recommended for most species, between 5 and 6 mg L-1, tilapia can tolerate low concentrations. Wambua, Home, Raude, and Ondimu. (2021) state that these animals can support oxygen levels between 0.4 and 0.7 mg L-1, but the development may be compromised. Although the dissolved oxygen is below the recommended for most species, between 5 and 6 mg L-1, tilapia can tolerate low concentrations. Wambua, Home, Raude, and Ondimu. (2021) state that these animals can support oxygen levels between 0.4 and 0.7 mg L-1, but the development may be compromised. High ammonia levels may affect osmoregulation, excretion, and oxygen transport; therefore, levels below 0.24 mg L-1 are recommended (Harsij, Kanani, & Adineh, 2020). Thus, the ammonia level found in our study is within the recommended range. High ammonia levels may affect osmoregulation, excretion, and oxygen transport; therefore, levels below 0.24 mg L-1 are recommended (Harsij, Kanani, & Adineh, 2020). Thus, the ammonia level found in our study is within the recommended range. The pH remained within the recommended range for the species, which are between 6.0 and 8.5 (Mili et al., 2023). The main reasons for pH alterations are breathing, fertilizing, liming, photosynthesis, and pollution. The pH value below 4.0 and above 9.0 stress animals, which can cause death (Makled, Hamdan, & El‐Sayed, 2019). Regarding the recovery of Bacillus sp SMIA-2 from the intestine (Figure 2), its presence was not detected in the treatment in which it was not added to the feed. This demonstrates that there was no contamination of aquaria or feed. Therefore, the bacterium was not present in the system. It was possible to recover the bacterium from the intestine in the other treatments. Pierro et al. using 0.1% peptone water (w/v), and 0.1 mL were seeded on the surface of Petri dishes containing tryptic soy agar (TSA). Dishes were incubated at 50°C for 48 hours for posterior counting of colonies. The results were expressed as the number of spores per mL. using 0.1% peptone water (w/v), and 0.1 mL were seeded on the surface of Petri dishes containing tryptic soy agar (TSA). Dishes were incubated at 50°C for 48 hours for posterior counting of colonies. The results were expressed as the number of spores per mL. Figure 1. Photomicrography of the intestine section of fish with the measurement of villus height. Figure 1. Photomicrography of the intestine section of fish with the measurement of villus height. Recovering Bacillus sp SMIA-2 from the intestine After euthanasia, fish were externally decontaminated with 70% alcohol by immersion in a glass container for five minutes. Intestines were removed, weighed, and macerated with a solution of 0.1% (w/v) sterile peptone water (Jatobá et al., 2008). Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 To counter SMIA-2 spores, samples were submitted to thermal shock (80ºC 10 min.-1) (Rabinovitch & Oliveira, 2015) in a water bath and immediately cooled in an ice bath. Then, serial dilutions were conducted Page 4 of 11 Pierro et al. Results and discussion Effect of adding different concentrations of Bacillus sp SMIA-2 in the diet on the zootechnical development and intestinal morphometry of tilapia juveniles Effect of adding different concentrations of Bacillus sp SMIA-2 in the diet on the zootechnical development and intestinal morphometry of tilapia juveniles The number of colony-forming units increased with increasing concentrations of SMIA-2 in the feed. Such results demonstrate their survival ability due to resistance of their spores to gastric and biliary acids. Therefore, they are viable as a probiotic. In tilapia post-larvae fed a feed containing between 5 and 10 g kg-1 Bacillus subtilis, between 4.74x105 CFU g-1 bacteria were recovered from the intestine (Tachibana et al., 2011). Acta Scientiarum Animal Sciences v 46 e64312 2024 In tilapia post-larvae fed a feed containing between 5 and 10 g kg-1 Bacillus subtilis, between 1.15x104 and 4.74x105 CFU g-1 bacteria were recovered from the intestine (Tachibana et al., 2011). Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Bacillus as additives in fish diets Page 5 of 1 Figure 2. Effect of different concentrations of Bacillus sp. SMIA-2 and its enzymes on the number of spores recovered from the intestines of tilapias. Page 5 of 11 Page 5 of 11 Page 5 of 11 Bacillus as additives in fish diets Figure 2. Effect of different concentrations of Bacillus sp. SMIA-2 and its enzymes on the number of spores recovered from the intestines of tilapias. Animal performance Among the variables statistically analyzed, those with significance were final weight (FW), weight gain (WG), food conversion ratio (FCR) (Table 2), individual feed consumption (IFC), survival rate (SR), specific growth rate (SGR), protein efficiency ratio (PER), viscera weight (VW), hepatosomatic index (HSI), and viscerosomatic index (VSI) (Table 3). Regarding final weight, weight gain, and food conversion, the animals had results statistically similar in all treatments. Table 2. Mean values of performance characteristics in tilapia juveniles fed different inclusion levels of SMIA-2. Inclusion levels of smia-2 (g kg-1 feed) Performance Characteristics FW (g) WG (g) FCR Control 19.656 18.952 1.237 0.383 23.859 23.156 1.121 0.765 19.380 18.678 1.339 1.147 20.305 19.601 1.186 1.53 20.687 19.984 1.209 Table 2. Mean values of performance characteristics in tilapia juveniles fed different inclusion levels of SMIA-2. At the end of the experimental period, the livers of the animals had a standard coloration and satisfying integrity. The increase in this organ size might be due to a low-protein but high-carbohydrate and high-fat diet, resulting in a yellowish-brown color. As the feed supplied to animals was the same in the five treatments, it is improbable that the liver increase is due to the feed quality. Thus, we suggest that the liver weight increase might be caused by the energy reserves of animals, which can be beneficial because the reserves have an essential role as a glucose source for animals in the pre- and postprandial period (Da Silveira, Logato, & Da Conceição Pontes, 2009). As Table 3 shows, the inclusion of 1.147 g kg-1 Bacillus sp. SMIA-2 in the feed promoted the best response regarding the variable LW. Regarding HPI, VW, and VSI, the concentration of SMIA-2 of 1.147 g kg-1 c presented a better response. Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Table 3. Mean values of liver weight, viscera weight, hepatosomatic index, and viscerosomatic index in tilapia juveniles fed different inclusion levels of Bacillus sp. SMIA-2. Inclusion levels OF SMIA-2 (g kg-1 of feed) Performance Characteristics LW (g) VW (g) HSI VSI Control 0.392ab 1.436abc 1.998ab 7.379a 0.383 0.362ab 1.666a 1.572ab 7.101a 0.765 0.242b 1.094c 1.253b 5.729a 1.147 0.408a 1.558ab 2.039a 7.701a 1.53 0.255ab 1.232bc 1.240b 5.969a Means followed by different letters (columns) statistically differ by Tukey’s test at 5% significance. Liver weight (LW), viscera weight (VW), hepatosomatic index (HSI), and viscerosomatic index (VSI). Histomorphometric analysis The statistical analysis evidenced that the villi height decreased with increasing levels of SMIA-2 in the feed, as illustrated in Figure 3. Figure 3. Intestinal villi height of tilapia according to different levels of SMIA-2 in the feed. Figure 3. Intestinal villi height of tilapia according to different levels of SMIA-2 in the feed. The feed without symbiotics (control) resulted in higher intestinal villi height. Such results are similar to those reported by Cechim et al. (2013), who investigated the addition of prebiotic mannan oligosaccharide (MOS) in the concentration of 4.0 g kg-1 to tilapia feed, and observed a reduction in intestinal villi height compared to the control. When environmental and sanitary conditions are favorable, the use of probiotics may have no effect (Dawood, Koshi, Abdel‐Daim, & Van Doan, 2019). As tilapia is rustic and easily adaptable, the action of additives may be masked. Animal performance alues of liver weight, viscera weight, hepatosomatic index, and viscerosomatic index in tilapia juveniles fed different inclusion levels of Bacillus sp. SMIA-2. Pierro et al. VSI is a factor that affects fish farming. An increase in fat deposition in the viscera can reduce the commercial value of fish (Wang et al., 2016). As tilapia is a species with high popularity and the second most farmed fish species globally (Islam, Rohani, & Shahjahan, 2021), the maximal use of the animal is desirable. According to Moraes et al. (2018), waste of fishing (head, entrails, skin, and dorsal spine) may represent 62.5% animal weight and cause environmental contamination. Thus, the lower the weight of viscera, the better the use by the animal. Page 7 of 11 Bacillus as additives in fish diets Bacillus as additives in fish diets ean values of performance characteristics in tilapia juveniles with the addition of different commercial symbiotics and Bacillus sp. SMIA-2. Table 4. Mean values of performance characteristics in tilapia juveniles with the addition of different commercial symbiotics and Bacillus sp. SMIA-2. Treatments Performance Characteristics IFC (g) FW (g) WG (g) FCR Control 26.801b 25.43b 24.29a 1.10a SMIA-2 35.617a 35.45a 34.31a 1.04a Commercial product 1 36.653a 35.39a 34.25a 1.07a Commercial product 2 36.315a 36.28a 35.15b 1.03a Means followed by different letters (columns) statistically differ by the Tukey test at 5% significance. Individual food consumption (IFC), final weight (FW), weight gain (WG), and feed conversion ratio (FCR). y different letters (columns) statistically differ by the Tukey test at 5% significance. Individual food consumption (IFC), final weight (FW), weight gain (WG), and feed conversion ratio (FCR). Table 5. Mean values of performance characteristics in tilapia juveniles fed different commercial symbiotics and Bacillus sp. SMIA-2. Treatments Performance Characteristics SR (%) SGR (%) PER (%) VW IL IC Control 64ab 6.716b 2.5109a 1.1416b 47.60b 5.8458b SMIA-2 42b 7.472a 2.6749a 2.0204ab 44.80b 5.6325b Commercial product 1 82a 7.457a 2.5909a 2.6609a 58.20a 7.3616a Commercial product 2 76a 7.529a 2.6881a 2.6798a 57.74a 7.2167a Means followed by different letters (columns) statistically differ by Tukey’s test at 5% significance. Survival rate (SR), specific growth rate (SGR), protein efficiency ratio (PER), viscera weight (VW), intestinal length (IL), and intestinal coefficient (IC). Table 5. Mean values of performance characteristics in tilapia juveniles fed different commercial symbiotics and alues of performance characteristics in tilapia juveniles fed different commercial symbiotics and Bacillus sp. SMIA-2. The specific growth rate (SGR) and the protein efficiency ratio (PER) presented similar results in the groups fed different symbiotics. In this sense, Bacillus sp. SMIA-2 was more similar to the commercial products as a growth-promoting additive than the control group. Marengoni et al. (2010) reported that SGR is directly proportional to the sanitary conditions of the medium. In this study, water quality indicators remained within the recommended range for the species. Therefore, they did not compromise the normal physiological activities of animals. Both in the treatments with commercial products and the treatments with Bacillus sp. SMIA-2, there was an efficient use of proteins by fish, suggesting that including symbiotics in feed influenced the animal metabolism. Page 7 of 11 Further, the major residues in the excretion of fish are phosphorus and nitrogen compounds, which in excessive quantities might degrade the farming environment, resulting in low water quality (Macedo & Sipaúba-Tavares, 2010). Thus, additives in the feed can increase digestibility and promote better use of nutrients, impeding the accumulation of nutrients in the water (Gomes et al., 2016). Although the viscera weight (liver, gallbladder, stomach, pancreas, and intestine) was higher with the inclusion of commercial products, its value was higher in the treatment with Bacillus sp. SMIA-2 than in the control group. Considering that the meat of fish is the main product of interest, fillet yield depends on many factors, including the percentage of residues. In this aspect, the lower the weight of entrails, fins, skin, and head, the higher the animal yield (De Moraes Gonçalves, De Almeida, & Santo Borges, 2003). Considering that tilapia is the most farmed fish species in Brazil, a lower viscera weight is desirable to produce fillets with a higher yield. Therefore, Bacillus sp. SMIA-2 has a slight advantage compared to the commercial products tested, considering that FW in the three treatments was statistically similar. Intestinal length (IL) and intestinal coefficient (IC) also presented an advantage for Bacillus sp. SMIA-2 compared to the commercial products and the control group. The IC of omnivorous animals ranges between 0.6 and 8.0, and animals with smaller intestines have a higher number of villi, increasing the surface, for example, with a higher development of pyloric caeca, expanding the absorption surface without increasing the IL (Ferreira et al., 2014). Thus, if it is intended to enhance the fillet yield of these animals, Bacillus sp. SMIA-2 has an advantage compared to the commercial products, although tests are still in an initial phase regarding aquatic animals. Effects of adding SMIA-2 and different commercial products in the diet on the zootechnical development of tilapia juveniles. Mean values of the quality parameters measured during the experimental period were temperature 26.94°C; dissolved oxygen – 3.87 mg L-1; ammonium 0.00205 mg L-1, and potential of hydrogen (pH) – 6.93. Regarding animal performance, the variables with statistical significance were final weight, weight gain, feed conversion ratio (Table 4), individual food consumption, survival rate, specific growth rate, protein efficiency ratio, viscera weight, intestine length, and intestinal coefficient (Table 5). According to the results, there was a significant effect on the individual food consumption, final weight, and weight gain when the symbiotics were added to the feed. In addition, feed conversion ratio did not present significant differences between the treatments, the groups with symbiotics showed a better response in these development characteristics. Such results corroborate Cornélio et al. (2013) and Pezzato, Menezes, Barros, Guimarães, and Schich (2006), which added probiotic bacteria to tilapia fish feed. Based on the principle that one of the action mechanisms of probiotics is the competition for absorption, its inclusion in the feed might have prevented the colonization by prejudicial bacteria, resulting in a better nutrient absorption efficiency. According to Marengoni et al. (2010), probiotics in the feed can decrease production costs because microorganisms can decompose macronutrients, transforming them into simpler compounds, which results in better food use. Although the use of bacteria from genus Bacillus in aquaculture can be related to good survival indexes, the survival rate with the use of Bacillus sp. SMIA-2 in this experiment was lower than the treatments with commercial symbiotics. To permit the symbiotics to express their beneficial effects, many factors must be considered, such as the presence of stressors, low water quality, high storage densities, or diseases (Silva, Salomao, Mareco, Dal Pai, & Santos al., 2021). Marengoni et al. (2010) highlight that it is not always possible to evidence the positive effects of additives as they may depend on the diet ingredients, the stress levels of animals, and each the characteristics of products. Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Conclusion The inclusion of Bacillus sp. SMIA-2 showed great potential as an additive for fish nutrition, even compared to the commercial products tested. It demonstrated the capacity to enhance the performance of animals and caused a significant increase in intestinal villi, contributing to better nutrient absorption and animal health. Pierro et al. Pierro et al. Figure 4. Intestinal villi height of fish according to the diet 1 (Control), 2 (SMIA-2), 3 (Commercial product 1), 4 (Commercial product 2). Figure 5. Photomicrography of the intestinal portion of a fish fed inclusion levels of Bacillus sp. SMIA-2 with the measurement of the villus height. Figure 5. Photomicrography of the intestinal portion of a fish fed inclusion levels of Bacillus sp. SMIA-2 with the measurement of the villus height. Such results are promising for fish nutrition, since there is an association between intestinal villi and the nutrition and health of animals. The inclusion of microorganisms in the diet can influence the intestinal microbiota of animals, acting in the villi necessary for intestinal functioning. Villi expand intestinal surface, enhancing water, ion, and nutrient absorption. Therefore, they have a key role in the development and health of animals, which may be altered according to the diet (de Souza & Ferreira, 2022). Intestinal histomorphometric analysis The height of intestinal villi of the fish was higher in the treatment with Bacillus sp SMIA-2 in the feed, as shown in Figures 4 and 5. Considering that the intestinal mucosa is more intact, the higher the villi and the nutrient absorption capacity (Zhaxi et al., 2020). Such results are promising for fish nutrition, because there is an association between intestinal villi importance and the nutrition and health of animals. Acta Scientiarum. Animal Sciences, v. 46, e64312, 2024 Page 8 of 11 Pierro et al. Figure 4. Intestinal villi height of fish according to the diet 1 (Control), 2 (SMIA-2), 3 (Commercial product 1), 4 (Commercial product 2). Page 8 of 11 References Andrade, M. V. V. D., Delatorre, A. B., Ladeira, S. A., & Martins, M. L. L. (2011). Production and partial characterization of alkaline polygalacturonase secreted by thermophilic Bacillus sp. SMIA-2 under submerged culture using pectin and corn steep liquor. 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https://openalex.org/W4289299504	https://zenodo.org/record/1481543/files/fraternail.pdf	English	null	Angular Momentum Accretion onto Disc Galaxies	Zenodo (CERN European Organization for Nuclear Research)	2,018	cc-by	3,199	Angular Momentum Accretion onto Disc Galaxies Filippo Fraternali1 and Gabriele Pezzulli2 1Kapteyn Astronomical Institute, University of Groningen, P.O. Box 800, 9700AV Groningen, The Netherlands, email: fraternali@astro.rug.nl 2Department of Physics, ETH Zurich, Wolfgang-Pauli-Strasse 27, 8093 Zurich, Switzerland email: gabriele.pezzulli@phys.ethz.ch Filippo Fraternali1 and Gabriele Pezzulli2 1Kapteyn Astronomical Institute, University of Groningen, P.O. Box 800, 9700AV Groningen, The Netherlands, email: fraternali@astro.rug.nl 2Department of Physics, ETH Zurich, Wolfgang-Pauli-Strasse 27, 8093 Zurich, Switzerland email: gabriele.pezzulli@phys.ethz.ch Abstract. Throughout the Hubble time, gas makes its way from the intergalactic medium into galaxies fuelling their star formation and promoting their growth. One of the key properties of the accreting gas is its angular momentum, which has profound implications for the evolution of, in particular, disc galaxies. Here, we discuss how to infer the angular momentum of the accreting gas using observations of present-day galaxy discs. We ﬁrst summarize evidence for ongoing inside-out growth of star forming discs. We then focus on the chemistry of the discs and show how the observed metallicity gradients can be explained if gas accretes onto a disc rotating with a velocity 20 −30% lower than the local circular speed. We also show that these gradients are incompatible with accretion occurring at the edge of the discs and ﬂowing radially inward. Finally, we investigate gas accretion from a hot corona with a cosmological angular momentum distribution and describe how simple models of rotating coronae guarantee the inside-out growth of disc galaxies. Keywords. Angular momentum, gas accretion, hot halo, metallicity gradient, corona Astronomy in Focus, Volume 1 XXXth IAU General Assembly, August 2018 Teresa Lago & Danail Obreschkow, ed. Astronomy in Focus, Volume 1 XXXth IAU General Assembly, August 2018 Teresa Lago & Danail Obreschkow, ed. c⃝2018 International Astronomical Union DOI: 10.5281/zenodo.1481543 1. Introduction At variance with massive quiescent ellipticals, which assembled most of their mass a long time ago and experienced, at some points in the past, an abrupt decline of their star formation rate, the majority of presently star forming galaxies have been undergoing, for most of the cosmic time, a rather constant or gently declining star formation history (e.g. Paciﬁci et al. 2016). This could in principle be explained either by a gradual consumption of a very large initial amount of cold gas, or by continuous accretion of new gas from the intergalactic medium. Both theory and observations strongly argue in favour of the second option, as i) gradual accretion is expected from the cosmological theory of structure formation (e.g. van den Bosch et al. 2014); ii) star forming galaxies have relatively short depletions times (Saintonge et al. 2011) and iii) preventing a huge initial reservoir of gas from very rapid exhaustion requires an implausibly low star formation eﬃciency, in stark contrast with observations (Kennicutt & Evans 2012; Fraternali & Tomassetti 2012). Observing gas accretion into galaxies directly has proven challenging (Sancisi et al. 2008; Rubin et al. 2012). In a galaxy like the present-day Milky Way gas accretion does not seem to take place in the form of cold gas clouds at high column densities, like the classical high-velocity clouds (Wakker & van Woerden 1997) as their estimated accretion rate is too low (Putman et al. 2012) and their origin may be, at least partially, from a galactic fountain rather than a genuine accretion (Fraternali et al. 2015, Fox et al. 2016). Lower column densities have been probed in absorption but the accretion rates are more uncertain (Lehner & Howk 2011; Tumlinson et al. 2017). A possibility is that 38 2 Filippo Fraternali & Gabriele Pezzulli Figure 1. Distribution of the observed speciﬁc radial growth rates (νR) of a sample of nearby galaxies (Pezzulli et al. 2015). Note how the vast majority are growing inside out still at z = 0. ◼ ◼ Filippo Fraternali & Gabriele Pezzulli Figure 1. Distribution of the observed speciﬁc radial growth rates (νR) of a sample of nearby galaxies (Pezzulli et al. 2015). Note how the vast majority are growing inside out still at z = 0. gas accretion takes place from the cooling of the hot gas, the galactic coronae that surround the Milky Way and similar galaxies (Miller & Bregman 2015). 1. Introduction The cooling can be stimulated by the mixing with the disc gas through fountain condensation (Armillotta et al. 2016; Fraternali 2017). Alternatively, it may come from cold cosmological ﬁlaments directly reaching the discs (e.g. Kereˇs et al. 2009). To distinguish between these scenarios it is crucial to estimate the properties of the accreting gas. A powerful tool to investigate the properties of the accreting gas is to infer them indirectly (backward approach) from observations of galaxy discs today. A simple example of this backward approach consists in the estimate of the accretion rates from the star formation histories (e.g. Fraternali & Tomassetti 2012). More elaborate estimates allow us to derive the angular momentum of the gas, the location where the accretion should take place and the properties of the medium from which the accretion originates. These topics are the focus of this proceeding. 3. Disentangling models of accretion We have seen that most spiral galaxies must have been (and probably are) gradually accreting angular momentum rich gas from the surrounding medium. The compelling question arises of what is the exact physical mechanism by which this happens. Two competing scenarios exist: cold mode accretion and hot mode accretion (e.g. Birnboim & Dekel 2003; Binney 2004) and diﬀerent modes can dominate at diﬀerent masses and redshift. In the former case, cold and angular momentum rich gas from intergalactic ﬁlaments joins the disc at large radii and then somehow drifts inwards to sustain star formation with the observed radial proﬁle throughout the disc. We can call this purely radial accretion. In the second scenario, instead, the gas accreting onto the halo does not join the main body of the galaxy directly, but it is rather stored (together with its angular momentum) into a hot CGM (corona) and then only gradually condenses on to the disc as a gentle ‘rain’, which may be modelled, at least at ﬁrst order, as mostly vertical accretion (perpendicular to the galaxy disc). ( ) With appropriate choices of the parameters, both scenarios can give rise to the same structural evolution of the disc, as constrained by observations of the star and gas content of galaxies as a function of galactocentric radius and time (Pezzulli & Fraternali 2016a). The two models however diﬀer enormously (and can thus be distinguished) in terms of chemical evolution. This is because vertical accretion of relatively metal-poor gas has a metal-dilution eﬀect, which goes in the direction of counter-acting metal enrichment by local star formation, whereas radial accretion implies that, before arriving at the position where it is ﬁnally locked in to stars, each gas element will have already traversed other regions of the galaxy, where it will have been chemically enriched by the stars being formed there. We emphasize that i) this observational test is better performed on gas- phase abundances of α elements (as this choice minimizes uncertainties due to stellar radial migration and time delays in chemical enrichment) and ii) this kind of comparison between models is only meaningful at ﬁxed structural evolution, as, otherwise, diﬀerences in other leading order eﬀects (gas fraction, star formation eﬃciency and so on) dominate over those due to diﬀerent geometry of accretion. With these speciﬁcations clariﬁed, the discriminating power of the method is remarkable. 2. Accretion of angular momentum on star forming galaxies A very important observational fact about the evolution of currently star forming spiral galaxies, is that they have been increasing in size while increasing in mass (inside-out growth, e.g. Larson 1976; Dale et al. 2016). This is most likely due to the fact that the gas that has been accreted most recently is more rich in angular momentum with the respect to the one which was accreted at earlier epochs, a very well established prediction of the cosmological theory of tidal torques (Peebles 1969). Crucially, observations indicate the radial growth – and therefore angular momentum accretion – is also a gentle process, which has been proceeding at a regular rate through- out galaxy evolution and is still ongoing today, as shown by studies of spatially resolved stellar populations (e.g. Williams et al. 2009; Gogarten et al. 2010) or recent star forma- tion (e.g. Mu˜noz-Mateos at al. 2007). Pezzulli et al. (2015) have proposed a quantitative analysis of the phenomenon. They have shown that, relative to the well known exponen- tial proﬁle of the stellar mass surface density of spiral galaxies, the radial proﬁle of the current star formation rate surface density shows a mild depletion in the inner regions and a slight enhancement in the outer ones, which agree both qualitatively and quanti- tatively with ongoing radial growth of stellar discs at a low but measurable rate. Figure 1 shows the distribution of the measured speciﬁc radial growth rate νR ≡˙R⋆/R⋆of the stellar scale-length R⋆of the sample of nearby spiral galaxies from that study. The vast 39 Angular Momentum Accretion onto Disc Galaxies majority of objects are currently growing, at a rate about equal to one third of their spe- ciﬁc star formation rate (sSFR, or νM ≡˙M⋆/M⋆≃0.1 Gyr−1 at z = 0, e.g. Speagle et al. 2014). Furthermore, the results were shown to agree quantitatively with expectations for gradual angular momentum assembly of galaxies evolving along the speciﬁc angular momentum versus stellar mass (Fall) relation (Fall & Romanowsky 2013). 3. Disentangling models of accretion This is illustrated in Figure 2, where the predictions are shown, for the abundance gradient of α-elements in the ISM of the Milky Way, for models with purely vertical, purely radial and mixed accretion. Details can be found in e.g. Pitts & Tayler (1989), Sch¨onrich & Binney (2009) and Pezzulli & Fraternali (2016a). The latter work proposes an analytic and general approach to the problem, which can be readily applied to any galaxy or structural evolution model. The clear result is that a combination of vertical and radial accretion is required to match the observed gradient (e.g. Genovali et al. 2015, marked here as a dashed line). This is actually not surprising, when angular momentum conservation is taken into account. A purely radial accretion, in fact, requires the angular momentum of the accreting gas to be transferred to some other not very well identiﬁed phase. On the other hand, purely vertical accretion is only possible if the material is accreted, at any radius, with exactly the angular momentum needed for local centrifugal balance, as any discrepancy would force the condensed gas to move radially within the disc after accretion (see also Mayor & 40 4 Filippo Fraternali & Gabriele Pezzulli Figure 2. Abundance proﬁles of gas phase α-elements, as a function of the galactocentric radius, for the Milky Way at z = 0 predicted by disc evolution models in which the gas accretes purely vertically, purely radially or through mixed (coupled) ﬂows. The observed proﬁle is shown by the dashed curve. The models are calculated as described in Pezzulli & Fraternali (2016b). ◼ ◼ Filippo Fraternali & Gabriele Pezzulli Filippo Fraternali & Gabriele Pezzulli Figure 2. Abundance proﬁles of gas phase α-elements, as a function of the galactocentric radius, for the Milky Way at z = 0 predicted by disc evolution models in which the gas accretes purely vertically, purely radially or through mixed (coupled) ﬂows. The observed proﬁle is shown by the dashed curve. The models are calculated as described in Pezzulli & Fraternali (2016b). Vigroux 1981; Bilitewski & Sch¨onrich 2012). We now discuss whether a coherent physical picture can naturally give account of the ﬁndings discussed so far. 4. A consistent picture The model which better reproduces the chemical evolution of the Milky Way (§3) requires gas accretion occurring with a speciﬁc angular momentum that is 75 ± 5% of that of the disc at each radius. This is very naturally expected for a hot mode accretion scenario (i.e. from the hot corona). Virtually every hydrodynamically consistent model of the hot CGM requires in fact the hot gas to be not in local centrifugal equilibrium, as the high temperatures will generally imply a signiﬁcant contribution of pressure support (in addition to rotation) against gravity. Note that the same model also predicts the presence of moderate radial ﬂows (a few km s−1 or less) within the disc: crucially, how- ever, this radial ﬂow is not due to equatorial accretion of cold ﬂows, but it is rather the natural consequence of the rotation lag of the accreting (hot) gas and angular momentum conservation. Two main questions arise to further test whether the model is viable in a cosmological context. First, if the disc accretes material with a local deﬁcit of angular momentum, can the accretion still globally provide enough angular momentum, as required to sustain the global radial growth of the disc (§2)? Second, is the implied rotation of the corona consistent with cosmological expectation from tidal torque theory? The answer to these questions requires building self-consistent models of the hydro- dynamical equilibrium of a hot rotating corona in a galaxy scale gravitational potential and with a given angular momentum distribution. Pezzulli, Fraternali & Binney (2017) described the solution to this problem and showed that a corona with a cosmologically motivated angular momentum distribution can naturally develop, in the proximity of the disc, rotation velocities close to the value required to match the chemical constraints. The model also predicts that the rotation velocity of the hot gas should drop signiﬁcantly when approaching the virial radius.The ﬁrst prediction is in excellent agreement with the recent observations by Hodges-Kluck et al. (2016); the second will require next-generation X-ray observations to be conﬁrmed or discarded. 4. A consistent picture Pezzulli, Fraternali & Binney (2017) also found that the speciﬁc angular momentum of 41 Angular Momentum Accretion onto Disc Galaxies the inner corona increases rather steeply with radius and becomes larger than the average angular momentum of the disc at a radius Rcrit slightly larger than the disc scale-length, but well within the range of direct contact between the galaxy and the hot halo. This is is suﬃcient to make the corona a plausible source of angular momentum growth, provided that the accretion of coronal gas is particularly eﬃcient at relatively large radii, as predicted for instance by models of fountain-driven condensation (e.g. Marasco et al. 2012; Fraternali et al. 2013), and/or that the accretion is inhibited or counter-acted in the very central regions by star formation or AGN feedback (as suggested for instance for the Milky Way by the discovery of the Fermi bubbles; Su et al. 2010). 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https://openalex.org/W3136398067	https://leopard.tu-braunschweig.de/servlets/MCRFileNodeServlet/dbbs_derivate_00048137/Borrero-de Acu%C3%B1a_fbioe-09-642023.pdf	English	null	Fed-Batch mcl- Polyhydroxyalkanoates Production in Pseudomonas putida KT2440 and ΔphaZ Mutant on Biodiesel-Derived Crude Glycerol	Frontiers in bioengineering and biotechnology	2,021	cc-by	8,184	BRIEF RESEARCH REPORT published: 16 March 2021 doi: 10.3389/fbioe.2021.642023 BRIEF RESEARCH REPORT published: 16 March 2021 doi: 10.3389/fbioe.2021.642023 Fed-Batch mcl- Polyhydroxyalkanoates Production in Pseudomonas putida KT2440 and 1phaZ Mutant on Biodiesel-Derived Crude Glycerol José Manuel Borrero-de Acuña1,2, Manfred Rohde3, Cesar Saldias4 and Ignacio Poblete-Castro5* 1 Institute for Microbiology, Technische Universität Braunschweig, Braunschweig, Germany, 2 Braunschweig Integrated Centre of Systems Biology (BRICS), Technische Universität Braunschweig, Braunschweig, Germany, 3 Central Facility of Microscopy, Helmholtz Centre for Infection Research, Braunschweig, Germany, 4 Departamento de Química Física, Facultad de Química y Farmacia, Pontiﬁcia Universidad Católica de Chile, Macul, Chile, 5 Biosystems Engineering Laboratory, Faculty of Life Sciences, Center for Bioinformatics and Integrative Biology, Universidad Andres Bello, Santiago, Chile Keywords: mcl-poly(3-hydroxyalkanoates), fed-batch, crude glycerol, pseudomonas putida, PHA depolymerase Received: 15 December 2020 Accepted: 18 February 2021 Published: 16 March 2021 Received: 15 December 2020 Accepted: 18 February 2021 Published: 16 March 2021 Edited by: Prasun Kumar, Chungbuk National University, South Korea Crude glycerol has emerged as a suitable feedstock for the biotechnological production of various industrial chemicals given its high surplus catalyzed by the biodiesel industry. Pseudomonas bacteria metabolize the polyol into several biopolymers, including alginate and medium-chain-length poly(3-hydroxyalkanoates) (mcl-PHAs). Although P. putida is a suited platform to derive these polyoxoesters from crude glycerol, the attained concentrations in batch and fed-batch cultures are still low. In this study, we employed P. putida KT2440 and the hyper-PHA producer 1phaZ mutant in two different fed-batch modes to synthesize mcl-PHAs from raw glycerol. Initially, the cells grew in a batch phase (µmax 0.21 h−1) for 22 h followed by a carbon-limiting exponential feeding, where the speciﬁc growth rate was set at 0.1 (h−1), resulting in a cell dry weight (CDW) of nearly 50 (g L−1) at 40 h cultivation. During the PHA production stage, we supplied the substrate at a constant rate of 50 (g h−1), where the KT2440 and the 1phaZ produced 9.7 and 12.7 gPHA L−1, respectively, after 60 h cultivation. We next evaluated the PHA production ability of the P. putida strains using a DO-stat approach under nitrogen depletion. Citric acid was the main by-product secreted by the cells, accumulating in the culture broth up to 48 (g L−1) under nitrogen limitation. The mutant 1phaZ amassed 38.9% of the CDW as mcl-PHA and exhibited a speciﬁc PHA volumetric productivity of 0.34 (g L−1 h−1), 48% higher than the parental KT2440 under the same growth conditions. The biosynthesized mcl-PHAs had average molecular weights ranging from 460 to 505 KDa and a polydispersity index (PDI) of 2.4–2.6. Here, we demonstrated that the DO-stat feeding approach in high cell density cultures enables the high yield production of mcl-PHA in P. putida strains using the industrial crude glycerol, where the fed-batch process selection is essential to exploit the superior biopolymer production hallmarks of engineered bacterial strains. Reviewed by: Suchada Chanprateep Napathorn, Chulalongkorn University, Thailand Victoria Eugenia Santos Mazorra, Complutense University of Madrid, Spain Correspondence: Ignacio Poblete-Castro ignacio.poblete@unab.cl Correspondence: Ignacio Poblete-Castro ignacio.poblete@unab.cl Specialty section: This article was submitted to Bioprocess Engineering, a section of the journal Frontiers in Bioengineering and Biotechnology INTRODUCTION A newly isolated soil strain from Thailand, Pseudomonas sp. ASC2, proved to synthesize 3.02 (g L−1) of mcl-PHAs in ﬂask experiments. In a high cell density fermentation, P. putida GO16 attained 19 (g L−1) biomass using the waste polyol, showing a speciﬁc PHA volumetric productivity of 0.13 (gPHA L−1 h−1) (Kenny et al., 2012). The worldwide manufacture of petrochemical plastics reaches over 359 million tons annually as these materials are essential in our current lifestyle (Poblete-Castro et al., 2020a; Tournier et al., 2020). Unfortunately, synthetic plastics are not prone to biodegradation and massively accumulate in the oceans and soil ecosystems (Zettler et al., 2013; Lau et al., 2020). Factors like temperature and radiation decompose the polymers in small particles (<5 mm), entering now into the food chain (Law and Thompson, 2014). This phenomenon is detrimental to preserving natural environments and human health (Smith et al., 2018), demanding actions to move toward a circular bioproduction economy. To this end, commercial substitutes of petrochemical plastics are the biodegradable poly(3-hydroxyalkanoates) (PHAs). These biopolymers possess physical and mechanical properties similar to oil-based plastics displaying thermal malleability and elasticity and having suitable breaking points for thermoforming (Laycock et al., 2013). The industrial sectors that exploit these biopolymers include food, textile, agriculture, biomedicine, and electronics (Raza et al., 2018). While the polymerization of PHAs in P. putida is well characterized and relies on the PHA synthase proteins PhaC1 and PhaC2, the not yet fully understood depolymerization process is a result in part of the enzymatic action of PHA depolymerase (PhaZ, PP_5004) (Arias et al., 2013). Diﬀerent metabolic stimuli govern gene regulatory crosstalk, with the polymerization vs. depolymerization of PHA kinetics still unclear (Karmann et al., 2017; Velázquez-Sánchez et al., 2020). The accumulating evidence suggests that the polymerization regulatory control depends on the type of carbon substrate (De Eugenio et al., 2010). Inactivation of the phaZ gene in P. putida KT2442 boosted mcl-PHA production of cells grown on fatty acids (Cai et al., 2009). Conversely, non-related PHA carbon sources, glucose or gluconate, did not yield higher PHA titers in this mutant. Glycerol-grown cells of a phaZ-lacking strain of KT2440 produced 36% more biopolymer than the wild type using crude glycerol (Poblete-Castro et al., 2014a). Using fatty acids as substrates, a phaZ minus strain of KT2440 formed more than 70 (g L−1) of unsaturated mcl-PHA in fed-batch cultures (Vo et al., 2015). RESULTS AND DISCUSSION p In the last decade, the biodiesel industry has generated large quantities of crude glycerol, an inevitable by-product resulting from the esteriﬁcation process of fatty acids (Garlapati et al., 2016). Given the high glycerol surplus, the product market price is continually dropping (Zhang et al., 2020), making it an attractive substrate to derive biochemicals (Kaur et al., 2020). Microbial catabolism of crude glycerol presents some challenges since the biodiesel by-product contains methanol, traces of diesel, and heavy metals (Mothes et al., 2007; Samul et al., 2014). Remarkably, Pseudomonas strains can endure these toxic materials (Poblete-Castro et al., 2017) and fuel glycerol metabolic products into PHA biosynthetic pathways enabling the synthesis of medium-chain length (mcl-PHAs) (Kenny et al., 2012; Pappalardo et al., 2014; Fu et al., 2015; Liu et al., 2018) and copolymers of short-co-medium-chain length polyesters (Orellana-Saez et al., 2019; Pacheco et al., 2019). Batch and fed- batch production of mcl-PHA is feasible using crude glycerol in Pseudomonas putida where the strain KT2440 formed 34% of the cell dry weight (CDW) as polyester with a ﬁnal product titer of 1.45 (g L−1) in 75 h (Poblete-Castro et al., 2014a). Citation: Borrero-de Acuña JM, Rohde M, Saldias C and Poblete-Castro I (2021) Fed-Batch mcl- Polyhydroxyalkanoates Production in Pseudomonas putida KT2440 and 1phaZ Mutant on Biodiesel-Derived Crude Glycerol. Front. Bioeng. Biotechnol. 9:642023. doi: 10.3389/fbioe.2021.642023 March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 1 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. INTRODUCTION These experiments laid the groundwork for challenging the production capacities of the phaZ-deﬁcient mutant in high cell density cultures. Here, we assessed diﬀerent feeding strategies for high titer PHA production in P. putida strains on industrial crude glycerol. The DO-stat fed-batch fermentation is best suited to bioconvert raw glycerol into the elastomer polyesters where the 1phaZ mutant and the parental KT2440 reached mcl-PHA speciﬁc volumetric productivities of 0.34 and 0.23 (g L−1 h−1), respectively. PHAs are naturally occurring inclusion bodies during nutrient imbalance formed in microbes’ cytosolic space and reported as energy reservoirs (Madison and Huisman, 1999) and essential elements to cope with diﬀerent stress agents (Obruca et al., 2017). A manifold of companies currently manufactures PHAs at a large scale (Borrero-de Acuña et al., 2017). However, industrial production of PHAs is costly, given high operational expenses because of the employed carbon substrate and downstream processing (Chen et al., 2020). Hence, to overcome these setbacks, waste materials arise as low-cost carbon substrates to sustain bacterial growth and renewable polyester production (Cesário et al., 2014; Nielsen et al., 2017; Borrero-de Acuña et al., 2019). Combining engineered microbial cell factories with high cell density fermentations is a robust approach to achieve elevated PHA productivities (Choi et al., 2020). Constant-Feeding PHA Synthesis Under Nitrogen Limitation P. putida KT2440 and a phaZ-deﬁcient mutant strain demonstrated in a previous study to synthesize eﬃciently mcl-PHAs from crude glycerol in batch cultures (Poblete- Castro et al., 2014a). We now challenged these natural polyester producers in the process of choice for the industrial production of PHAs, the fed-batch culture. Applying diﬀerent feeding strategies using industrial crude glycerol without any modiﬁcation (Cremer Oleo, GmbH, Germany), we developed a three-stage fermentation where the two initial phases aimed to form biomass and the third stage mcl-PHA under nitrogen depletion. The batch cultures started with a biomass concentration of 0.13 (g L−1) in a 4 L working volume bioreactor. The culture broth initially contained 20 (g L−1) crude glycerol and 1 (g L−1) of glucose to prevent a characteristic extended lag phase (Escapa et al., 2012; Beckers et al., 2016), usually taking more than 10 h of P. putida cells growing on glycerol as the sole C source (Poblete-Castro et al., 2020b). As the P. putida cells propagate, foam developed March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 2 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. FIGURE 1 \| Constant-feeding fed-batch mcl-PHA production. The fermentation process comprised three phases: (i) batch, (ii) exponential feeding, and (iii) constant feeding of crude glycerol (50 g h−1) under nitrogen depletion. Time proﬁle of (A) P. putida KT2440 and (B) phaZ-knockout mutant. The data represent the mean values and standard deviation from two independent experiments. FIGURE 1 \| Constant-feeding fed-batch mcl-PHA production. The fermentation process comprised three phases: (i) batch, (ii) exponential feeding, and (iii) constant feeding of crude glycerol (50 g h−1) under nitrogen depletion. Time proﬁle of (A) P. putida KT2440 and (B) phaZ-knockout mutant. The data represent the mean values and standard deviation from two independent experiments. mutant from ∼50 (g L−1) to 45.4 and 46.7 (g L−1), respectively, a common trend of cells enduring overﬂow metabolism (Xu et al., 1999; Poblete-Castro et al., 2014b). At the end of the fermentation process (60 h), the wild-type KT2440 synthesized 9.7 (gPHA L−1) with a biopolymer content of 21.4% of the CDW (Figure 1A and Table 1), while the 1phaZ deletion mutant achieved 12.7 (gPHA L−1), amassing 27.2% of the cell biomass as polyester (Figure 1B and Table 1). Constant-Feeding PHA Synthesis Under Nitrogen Limitation Finally, the monomeric composition of the generated biopolymers by constant-feeding strategy was dominated by 3-hydroxydecanoate with a 75.4 and 76.1% relative molar fraction in the wild-type and 1phaZ mutant, respectively (Table 1). The rest of the monomers had the following decreasing proportion within the polymeric chain in both tested strains: 3-hydroxyoctanoate, 3-hydroxydodecanoate, 3-hydroxy-5-cis-dodecanoate, 3-hydroxyhexanoate, and 3-hydroxytetradecanoate. in the bioreactor (at 3 h), which dispersed once we provided antifoam (200 µL L−1). After 22-h cultivation, P. putida KT2440 and 1phaZ mutant had similar maximum speciﬁc growth rates of 0.21 h−1 (Figure 1A), reaching a CDW of 11.5 g L−1 (Figures 1A,B). Then, we started feeding glycerol exponentially, setting the speciﬁc growth rate at nearly 50% of µmax (0.1 h−1) (Figure 1). During this phase, no glycerol accumulated in the culture broth or the cells secreted by-products due to the coupled catabolism and anabolism under carbon limiting conditions (Poblete-Castro et al., 2012; González-Cabaleiro et al., 2015). At 40 h cultivation, we no longer provided glycerol and ammonium; we instead provided a substrate pulse to attain 10 (g L−1) of glycerol within the bioreactor. This procedure triggered nitrogen limitation, and once glycerol was under the detection limit (after 1 h), we fed the substrate, but this time at a constant mass ﬂow rate of 50 (g h−1). During the biopolymer production phase, we upheld the airﬂow rate, providing only ﬁlter air instead. The DO evolution showed a marked increase as the cells accumulated higher amounts of PHAs through the process, showing a reduced O2 demand as the cells no longer duplicate (Figures 1A,B). The P. putida strains secreted citric acid as the main co-product as glycerol accumulated in the medium, showing a ﬁnal yield on the polyol of 0.18 ± 0.01 (g g−1) and 0.16 ± 0.02, for the wild-type and the mutant strain, respectively (Table 1 and Figure 1). Indeed, nitrogen depletion of glycerol-grown P. putida cells inhibits the TCA cycle enzymes like isocitrate dehydrogenase (ICD), slowing the carbon ﬂux through the oxidative route (Beckers et al., 2016). The biomass suﬀered a slight reduction in KT2440 and the phaZ-knockout PHA Production Using a Dissolved-Oxygen-Stat Feeding (DO-Stat) Under Nitrogen Limitation As a response to glycerol exhaustion, the DO saturation increased; thus, every time the dissolved oxygen exceeded a value of 70%, an automated addition of the substrate (20 g L−1 glycerol) occurred in an interval of 15 min (Figure 2). This feeding operation did not reduce the formed biomass, and the intracellular polyester synthesis boosted, maintaining a rapid pace of accumulation until the termination of the process (Figure 2). After 20 h of feeding driven by the DO response, the wild-type strain attained a biomass titer of 49.5 (gCDW L−1), of which 13.8 (g L−1) consisted of mcl-PHA (Figure 2A and Table 1). Similarly, the depolymerase-deﬁcient mutant achieved a biomass yield of 52.4 (gCDW L−1) but comprised an enhanced biopolymer yield of 20.4 (g L−1)—overall PHA content 38.9%wt (Figure 2B). The speciﬁc PHA volumetric productivity (0.34 g L−1 h−1) reached by the 1phaZ mutant is the highest reported today to derive mcl-PHA using industrial crude glycerol (Table 1). P. putida KT2440 and its phaZ knockout mutant strain secreted 48 (g L−1) of citric acid in the DO-stat stage (Figure 2). However, citrate yields on glycerol obtained in the fed-batch cultivations were lower (Table 1) than the values previously reported (0.5 g g−1) in batch cultures (Poblete-Castro et al., 2014a). The co- production of citrate not only is detrimental in terms of carbon loss for biopolymer synthesis but also impacts negatively to the PHA production process as production costs boost, and the addition of the base solution to maintain the optimal pH exerts a dilution eﬀect lowering the PHA volumetric productivities (Warnecke and Gill, 2005). It is clear that further metabolic engineering eﬀorts must avoid the entry of the carbon ﬂux into the TCA cycle during the biopolymer production phase to diminish organic acid formation in P. putida KT2440. There are several routes to achieve this, beginning with the overexpression of acetyl-CoA carboxylase (ACC), which converts acetyl-CoA into malonyl- CoA, where the latter is the main precursor for PHA synthesis of substrates metabolized through the central carbon metabolism (Rehm et al., 1998; Poblete-Castro et al., 2013). PHA Production Using a Dissolved-Oxygen-Stat Feeding (DO-Stat) Under Nitrogen Limitation Despite the sound PHA volumetric concentration obtained in this study operating the constant-feeding scheme, the P. putida strains did not amass the PHA content previously reported in batch cultures (Poblete-Castro et al., 2014a). Thus, to fully harness the biopolymer production capacities of the 1phaZ mutant and the wild type, a dissolved oxygen-stat (DO-stat) feeding approach was applied. This well-controlled strategy has proven to yield higher PHA amount and content in engineered March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 3 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. TABLE 1 \| Medium-chain length (mcl-PHA) production in Pseudomonas strains under various fermentation modes utilizing crude glycerol. Strain Production mode Biomass PHA PHA content YCIT/Gly Citrate yield YPHA/Gly PHA yield Speciﬁc PHA volumetric productivity Monomeric composition (%) References (g L−1) (g L−1) (%wt) (g g−1) (g g−1) (g L−1 h−1) C6 C8 C10 C12 C12:1 C14 P. putida KT2440 Constant feeding 45.3 9.7 21.4 0.18 0.08 0.16 1.2 15.3 75.4 5.7 2.4 0.3 This study 1phaZ Constant feeding 46.7 12.7 27.2 0.16 0.10 0.21 1.3 16.1 76.1 5.9 0.6 N.D. This study P. putida KT2440 DO-stat 49.5 13.8 27.9 0.36 0.11 0.23 0.8 15.8 73.3 5.6 4.5 0.4 This study 1phaZ DO-stat 52.4 20.4 38.9 0.32 0.13 0.34 1.2 16.7 76.2 5.4 0.5 N.D. This study P. putida GO16 Fed batch 19.0 6.3 33.2 N.D. N.S 0.13 3 18 35 13 15 7 Kenny et al., 2012 Pseudomonas sp. ASC2 Batch 10.7 3.0 28.2 N.D N.S. 0.04 N.D. 4.8 N.D. N.D. 95.2 N.D. Muangwong et al., 2016 P. putida strains on glucose and aromatics (Poblete-Castro et al., 2014b; Borrero-de Acuña et al., 2020). For biomass formation, we repeated the same growth conditions as those employed during the constant-feeding approach, where a mixture of pure oxygen and air was provided to avoid oxygen limitation. Nitrogen was always suﬃcient at this stage, reaching both strains biomass productions of nearly 50 (g L−1) (Figure 2). At this point, we provided a pulse of glycerol (10 g L−1) to enable the cells to consume the remaining ammonium, and to get a better DO response, we provided only ﬁlter air as carried out in the constant-feeding cultivations. Likewise, citrate began to accumulate in the culture broth due to the metabolic shift provoked by nitrogen limitation (Figure 2). PHA Production Using a Dissolved-Oxygen-Stat Feeding (DO-Stat) Under Nitrogen Limitation Another path is to block the entry of acetyl-CoA into the Krebs cycle by inactivating in vivo the citrate synthase enzyme using RNA interference or antisense RNA (Desai and Papoutsakis, 1999; Ko et al., 2020), as the genetic systems can be activated during the nitrogen-limiting phase, which may have no negative impacts on biomass formation in the initial nutrient-suﬃcient phase. Finally, inspection of the monomer composition of the biosynthesized PHA highly resembled the previous feeding regime’s values. The March 2021 \| Volume 9 \| Article 642023 4 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. FIGURE 2 \| DO-stat fed-batch mcl-PHA production. The culture comprised three phases: (i) batch, (ii) exponential feeding, and (iii) DO-stat response for substrate feeding under nitrogen limitation. Time proﬁle of (A) P. putida KT2440 and (B) phaZ-knockout mutant. The data represent the mean values and standard deviation from two independent experiments. FIGURE 2 \| DO-stat fed-batch mcl-PHA production. The culture comprised three phases: (i) batch, (ii) exponential feeding, and (iii) DO-stat response for substrate feeding under nitrogen limitation. Time proﬁle of (A) P. putida KT2440 and (B) phaZ-knockout mutant. The data represent the mean values and standard deviation from two independent experiments. most abundant hydroxy acid encountered in both strains was the C10 (Table 1). This monomer’s relative molar fraction in the parental strain was 73.3%, whereas the 1phaZ knockout mutant presented 76.2% (Table 1). granules than those cultivated on pure glycerol (Poblete-Castro et al., 2014a), which are more unevenly shaped (Figures 3A–D). The PHA morphology depends on various factors, including the level of expression of granular associate proteins like phasins and PHA polymerase and depolymerase (Jendrossek, 2009), which in turn rely on the culture conditions and the growth substrate (De Eugenio et al., 2010). Under stress, PHA-producing bacteria alter the PHA content and its morphology as a response mechanism improving the survival rate when cells thrive under high salt concentrations and low and elevated temperatures or encountering toxic compounds (Obruca et al., 2020). As raw glycerol contains methanol and heavy metal, this could explain the intracellular granule’s observed alterations. Visualization of the Biosynthesized mcl-PHA and Physical Properties putida KT2440 constant feeding, (B) phaZ-knockout mutant constant feeding, (C) P. putida KT2440 DO-stat, and (D) phaZ-knockout mutant DO-stat. (E) Thermal properties of the biosynthesized mcl-PHA: Transition glass temperature (Tg), melting temperature (Tm), and crystallization temperature (Tc). Molecular weight (Mw), average-number molecular weight (Mn), and polydispersity index (PDI). The values represent mean and standard deviation from two replicates. FIGURE 3 \| Transmission electron micrographs of P. putida cells in fed-batch fermentation taken at 60 h fermentation. (A) P. putida KT2440 constant feeding, (B) phaZ-knockout mutant constant feeding, (C) P. putida KT2440 DO-stat, and (D) phaZ-knockout mutant DO-stat. (E) Thermal properties of the biosynthesized mcl-PHA: Transition glass temperature (Tg), melting temperature (Tm), and crystallization temperature (Tc). Molecular weight (Mw), average-number molecular weight (Mn), and polydispersity index (PDI). The values represent mean and standard deviation from two replicates. FIGURE 3 \| Transmission electron micrographs of P. putida cells in fed-batch fermentation taken at 60 h fermentation. (A) P. putida KT2440 constant feeding, (B) phaZ-knockout mutant constant feeding, (C) P. putida KT2440 DO-stat, and (D) phaZ-knockout mutant DO-stat. (E) Thermal properties of the biosynthesized mcl-PHA: Transition glass temperature (Tg), melting temperature (Tm), and crystallization temperature (Tc). Molecular weight (Mw), average-number molecular weight (Mn), and polydispersity index (PDI). The values represent mean and standard deviation from two replicates. FIGURE 3 \| Transmission electron micrographs of P. putida cells in fed-batch fermentation taken at 60 h fermentation. (A) P. putida KT2440 constant feeding, (B) phaZ-knockout mutant constant feeding, (C) P. putida KT2440 DO-stat, and (D) phaZ-knockout mutant DO-stat. (E) Thermal properties of the biosynthesized mcl-PHA: Transition glass temperature (Tg), melting temperature (Tm), and crystallization temperature (Tc). Molecular weight (Mw), average-number molecular weight (Mn), and polydispersity index (PDI). The values represent mean and standard deviation from two replicates. lacking the phaZ gene presented the same molecular weights and thermal properties of the accumulated PHA as the parental strain (Solaiman et al., 2003). Further, given the high polydispersity index values (PDI > 2), the synthesized mcl-PHA on crude glycerol would also perform appropriately in extruders as the elastomers exhibit narrow average-number molar mass distribution (Mn, 190 KDa), where these values are relatively close to commercial polyesters (Debuissy et al., 2017). While extracting the PHA from the cells, we could appreciate the elastomer ﬁlm that remained after chloroform evaporation in a Petri dish for puriﬁcation. Film materials are paramount in the packaging and agroindustry sectors. P. putida Strains The wild-type P. putida KT2440 (DSM 6125) was obtained from the DSMZ collection, Germany, and the 1phaZ mutant was constructed in a previous study (Poblete-Castro et al., 2014a). These strains were used for the diﬀerent fed-batch fermentations. Visualization of the Biosynthesized mcl-PHA and Physical Properties A ﬁlmable mcl-PHA was synthesized by Pseudomonas mediterranea using raw glycerol (Pappalardo et al., 2014). The obtained mcl-PHA presented nearly the same proportion of C10 in the monomeric chain than the ﬁlmable biopolymer, and above all, both elastomers display Tm values higher than 50◦C and Mw of 400 KDa. by 1phaZ knockout mutant (0.34 g L−1 h−1) is a step further in the quest to derive mcl-PHA from the polyol of the biodiesel industry in a more cost-eﬀective fashion. There is still room for improvement concerning PHA production since the P. putida cells secreted high citric acid levels. Further metabolic engineering endeavors must reduce the carbon wastage and redirect carbon ﬂux toward polyester biosynthetic pathways to mining the cell factory’s PHA production performance. Visualization of the Biosynthesized mcl-PHA and Physical Properties y p Crude glycerol contains harmful compounds, including methanol, salts, and heavy metals (Mothes et al., 2007; Samul et al., 2014). These elements impair cell growth and evoke stress responses at the proteome and transcriptome levels (Manara et al., 2012; Fu et al., 2015; Bojanoviˇc et al., 2017). This toxic feedstock might also inﬂuence cell morphology and the polymerization process of the polyesters. Figure 3 depicts micrographs acquired by transmission electron microscopy taken at the maximum PHA formation point (60 h) in the constant feeding (3A, KT2440 and 3B, 1phaZ mutant) and DO-stat fermentations (3C, KT2440 and 3D, 1phaZ mutant). Neither the parental strain nor the phaZ-disrupted mutant displays signiﬁcant cellular morphological variance or altered PHA inclusion bodies (Figure 3). Conversely to a previous study on crude glycerol, the cells showed no aggregation (Poblete-Castro et al., 2014a), indicating robust growth and proper mixing. Impurities present in the substrate source as black dots were distinctive in both growing P. putida strains. Notably, intracellular PHA structures had no apparent alteration by deleting the polymer disassembling enzyme PhaZ (Figures 3B,D). As previously noted, P. putida strains grown on crude glycerol presented larger numbers of intracellular PHA We also unveiled the thermal properties of the biosynthesized mcl-PHA via diﬀerential scanning calorimetry (DSC) of puriﬁed biopolymer sampled at 60-h cultivation from the bioreactors. The obtained glass transition points (Tg) were very similar among the produced mcl-PHAs (−10.5 to −11.9◦C) showing a single Tg characteristic of a copolymer of medium-chain length polyesters (Cheema et al., 2012; Pacheco et al., 2019; Figure 3E). As distinctive elastomers, the melting endotherms were in the range of 56.7–59.3◦C (Chan Sin et al., 2010; Gumel et al., 2012). Most importantly, the molecular weight of the puriﬁed PHA was higher than 400 KDa (Figure 3E), a prerequisite for processing these kinds of macromolecules industrially (Mothes et al., 2007), especially as softener materials (Muangwong et al., 2016; Liu et al., 2018). Together, the PHA thermal and physical properties did not vary signiﬁcantly between the polyesters synthesized by wild-type and the phaZ minus mutant (Figure 3E). A Pseudomonas resinovorans strain March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 5 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. FIGURE 3 \| Transmission electron micrographs of P. putida cells in fed-batch fermentation taken at 60 h fermentation. (A) P. PHA Characterization and Quantiﬁcation PHA Characterization and Quantiﬁcation The polyesters were ﬁrstly methanolized in order to determine the PHA monomeric compositions and the intracellular PHA content. For this, 10 ml of culture was transferred to a falcon tube and cells were harvested at 9,000 × g for 10 min at 4◦C (Eppendorf 5810 R, Hamburg, Germany). Pellets were washed once with distilled water. The supernatants were discarded, and the pelleted cells were stored at −20◦C until needed. The methanolysis procedure was performed as previously speciﬁed (Borrero-de Acuña et al., 2014). Gas chromatography (GC) coupled with mass spectrometry (MS) was used to analyze the methyl esters of monomers. One milliliter of the organic phase was injected into a Varian GCMS system 450GC/240MS ion trap mass spectrometer (Varian Inc., Agilent Technologies) at a split ratio of 1:10. The software employed to process the resulting data was the MS Workstation 6.9.3 (Varian Inc., Agilent Technologies). The diﬀerent compounds, i.e., the methyl esters of 3-hydroxyexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecanoate, 3-hydroxy-5-cis-dodecanoate, and 3- hydroxytetradecanoate, were split by using a FactorFour VF- 5ms capillary column (30 m × 0.25 mm i.d. × 0.25 mm ﬁlm thickness), including calibration with commercial PHB (Sigma−Aldrich, MI, United States) and puriﬁed mcl−PHA from a previous work (Oliva-Arancibia et al., 2017). The carrier gas helium was set to a ﬂow rate of 0.9 ml min−1. The temperatures for the injector and transfer line were established at 275 and 300◦C, respectively. The oven temperature was stepwise programmed as follows: 40◦C for 2 min, rising progressively from 40 to 150◦C at a rate of 5◦C min−1 and ultimately increasing to 280◦C at a constant rate of 10◦C min−1. To capture positive ions, an electron ionization at 70 eV was settled, while the mass spectra were registered by scanning ions of m/z 50 to m/z 650. The PHA concentration was determined by the method described by Lageveen et al. (1988). The percentage of biopolymer in relation with the CDW in two biological replicates was averaged out to ascertain PHA content (wt%). The fed batches were seeded with the second pre-culture to attain an initial cell density of (OD600 0.26). The fed- batch reactor contained M9 medium supplemented with 20 g L−1 crude glycerol, 0.12 g L−1 MgSO4·7H2O, and 8 ml of the trace element solution. CONCLUSION Strains were stored in 25% glycerol at −80◦C as glycerol stocks. Cells were routinely streaked onto Luria–Bertani (LB) agar plates and grown overnight to isolate single colonies. For any shake ﬂask cultivation, P. putida strains were aerobically grown at 180 rpm and 30◦C. Single colonies were picked from the plate and transferred into a 50 mL shake ﬂask containing 10 mL of LB liquid medium. Deﬁned minimal medium (M9) containing 6 (g L−1) crude glycerol (Cremer Oleo, GmbH, Hamburg, Germany) was employed for subsequent pre-culture cultivation. The industrial glycerol contains 80% glycerol, 0.5% methanol, 10% ash, 3% organic matter, and 6.5% water. The M9 medium composition In this study, we demonstrated that the applied feeding strategy during mcl-PHA synthesis in P. putida strains using the crude glycerol as substrate inﬂuences the amount of formed polyester but not their physical properties. We proved that a DO-stat fed-batch process is a more suitable feeding strategy than the constant-feeding approach to metabolize the toxic crude glycerol resulting in nearly 50% more biopolymer at the end of the fermentation in the phaZ-deﬁcient mutant and the wild-type KT2440. The attained speciﬁc PHA volumetric productivity March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 6 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. consisted (per liter) of 12.8 g Na2HPO4·7H2O, 3 g KH2PO4, 4.7 g (NH4)2SO4, and 0.5 g NaCl. After autoclave sterilization, the medium was supplemented with ﬁltered trace elements [6.0 FeSO4·7H2O, 2.7 CaCO3, 2.0 ZnSO4·H2O, 1.16 MnSO4·H2O, 0.37 CoSO4·7H2O, 0.33 CuSO4·5H2O, and 0.08 H3BO3 (mg L−1)] and 0.12 g of MgSO4·7H2O. Ten milliliters of M9 medium in a 50 mL shake ﬂask was inoculated with the overnight LB- grown culture at an initial OD at 600 nm of 0.2 and incubated overnight. A second pre-culture was initiated by transferring a predetermined volume of the previous one into 300 mL of M9 medium containing 10 g L−1 crude glycerol in 1 L baﬄed Erlenmeyer ﬂasks and cultivated overnight. was dried to constant weight at 100◦C. A photometric test (LCK 303 kit, Hach Lange, Danaher, United States) served to measure oﬄine the ammonium levels in the supernatant. Supernatant samples were withdrawn and accordingly diluted to analyze the crude glycerol and organic acid (citrate, isocitrate, succinate, fumarate, malate, pyruvate, and oxaloacetate) concentrations by HPLC Agilent 1260 (Agilent, Krefeld, Germany). PHA Characterization and Quantiﬁcation Overall, 4 L of working volume was set up in a 15 L vessel (B10 stirred tank bioreactor, Biologische Verfahrenstechnik, Basel, Switzerland) to conduct the fermentation processes. The airﬂow rate was maintained at 10 L min−1 (air-to-pure oxygen ratio was set to 10:1) during the biomass production phase. In the PHA production phase, the airﬂow rate was kept, but pure oxygen was no longer needed, and the reactor was sparged only with compressed air, providing better DO response when the carbon substrate was supplied. The temperature was set at 30◦C and 12.5 (w/v) of NH4OH was added as required to stabilize pH to 6.8 ± 0.1 during the course of the biomass formation phase and as nitrogen supply to avoid N limitation. Then, in the PHA production phase, the base was replaced by NaOH 10% (w/v). When required, Tego Antifoam (Evonik, Germany) was supplemented to prevent foaming (200 µL L−1). The agitation speed was automatically adjusted to 800 rpm in order to keep the dissolved oxygen level above 20% air saturation. The feeding solution contained per liter: 780 g crude glycerol and 12 g MgSO4·7H2O. An exponential feeding strategy was applied during the biomass production phase, following an exponential function (Eq. 1). F(t) = µset(V0.X0)e(µ set.t)(S0.Yxs)−1 (1) (1) Hereby, F is the feed rate (L h−1), µset is the desired speciﬁc growth rate (set to 0.1 h−1), S0 is the substrate concentration of the feed medium (780 g L−1 crude glycerol), t is the time after feed start (h), Yxs is the biomass yield on crude glycerol taken from Poblete-Castro et al. (2014a), V0 is the initial volume of the culture (L), and X0 is the initial biomass level (g cells L−1). CONCLUSION The HPLC system was equipped with an 8-mm Rezex ROA-organic acid H column (Phenomenex, United States), which was operated at 65◦C. The mobile phase consisted of 0.013 N H2SO4 at a 0.5 ml min−1 ﬂow coupled with a RID detector system (Agilent serie1260). AUTHOR CONTRIBUTIONS The glass transition, crystallization, and melting temperatures (Tg, Tc, and Tm, respectively) of each sample were determined by a Mettler–Toledo DSC 821e. The following ﬁve cycles were performed: (i) a ﬁrst heating from −40 to 200◦C at 10◦C min−1, (ii) an isotherm for 3 min, (iii) a cooling from 200 to −40◦C at 10◦C min−1, (iv) an isotherm for 3 min, and (v) a second heating from −40 to 200◦C at 10◦C min−1. IP-C conceived and performed the fed-batch fermentation and analytics. CS performed the thermal analysis. MR carried out the TEM studies. JB-dA and IP-C interpreted the data and wrote the manuscript. All authors contributed to the article and approved the submitted version. DATA AVAILABILITY STATEMENT The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s. FUNDING IP-C acknowledges ﬁnancial support from the projects INACH RG_17_19 and ANID-PIA-Anillo INACH ACT192057. We acknowledge the German Research Foundation and the Open Access Publication Funds of the Technische Universität Braunschweig. The weight average molecular weights (Mw) and the respective polydispersity indices of the samples were determined by size exclusion chromatography (SEC) along with a static light scattering Dawn EOS in line with an Optilab DSP interferometric refractometer (both were obtained from Wyatt Transmission Electron Microscopy Prior to ﬁxation, the bacteria were cooled down to 4◦C. Next, 2% of glutaraldehyde and 5% of formaldehyde (5%) were added. Cells were subsequently washed with cacodylate buﬀer (0.01 mol L−1 cacodylate, 0.01 mol L−1 CaCl2, 0.01 mol L−1 MgCl2 6H2O, and 0.09 mol L−1 sucrose, pH 6/9) and stained for The optical density (OD600 nm) was registered over time in a spectrophotometer (Ultraspec 2000; Hitachi, Japan) to determine cell growth. Ten milliliters of cells was harvested at 9,000×g for 10 min at 4◦C and washed once with distilled water prior to CDW gravimetric quantiﬁcation in pre-weighed tubes. The cell pellet March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org 7 Fed-Batch mcl-Polyhydroxyalkanoates Production on Crude Glycerol Borrero-de Acuña et al. 1 h at room temperature with 1% aqueous osmium solution. Dehydration was achieved by adding acetone at increasing concentrations (10, 30, 50, 70, 90, and 100%) and incubating the samples for 30 min at each time. Solely the dehydration with 70% acetone containing 2% uranyl acetate was allowed overnight. The Spurr formula for hard resin was applied to inﬁltrate samples with an epoxy resin. A diamond knife was used to slice the samples into ultra-thin sections, which were further counterstained with a mixture of uranyl acetate and lead citrate. A TEM910 transmission electron microscope (Carl Zeiss, Oberkochen, Germany) was operated at an acceleration voltage of 80 kV to acquire images. 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March 2021 \| Volume 9 \| Article 642023 Frontiers in Bioengineering and Biotechnology \| www.frontiersin.org REFERENCES doi: 10.1007/s40572-018-0206-z Conﬂict of Interest: The authors declare that the research was conducted in the absence of any commercial or ﬁnancial relationships that could be construed as a potential conﬂict of interest. Conﬂict of Interest: The authors declare that the research was conducted in the absence of any commercial or ﬁnancial relationships that could be construed as a potential conﬂict of interest. Solaiman, D. K. Y., Ashby, R. D., and Foglia, T. A. (2003). Eﬀect of inactivation of poly(hydroxyalkanoates) depolymerase gene on the properties of poly(hydroxyalkanoates) in Pseudomonas resinovorans. Appl. Microbiol. Biotechnol. 62, 536–543. doi: 10.1007/s00253-003-1317-4 Copyright © 2021 Borrero-de Acuña, Rohde, Saldias and Poblete-Castro. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Copyright © 2021 Borrero-de Acuña, Rohde, Saldias and Poblete-Castro. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Tournier, V., Topham, C. M., Gilles, A., David, B., Folgoas, C., Moya-Leclair, E., et al. (2020). 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https://openalex.org/W2991637143	https://www.itm-conferences.org/articles/itmconf/pdf/2019/07/itmconf_crimico2019_15026.pdf	English	null	Universal mathematical model of SAR signals for natural surfaces	ITM web of conferences	2,019	cc-by	3,511	, (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 https://doi.org/10.1051/itmconf /20193015026 Universal mathematical model of SAR signals for natural surfaces Alexander Bokov, Yury Markov, Vladimir Vazhenin, and Sergey Margilevsky Ural Federal University, Yekaterinburg, Russian Federation Alexander Bokov, Yury Markov, Vladimir Vazhenin, and Sergey Margilevsky Ural Federal University, Yekaterinburg, Russian Federation Abstract. The implementation of the basic version of the universal mathematical model of the formation of reflected signals for various airborne radar systems, including the synthetic antenna aperture radar, when working on natural surfaces are discussed in the article. A number of parameters are required to study, which must be taken into account for the correct modeling of echo signals, which must then be processed to calculate radar images. A method for constructing such a mathematical model in MATLAB is presented with some modeling results for a homogeneous rough surface and two typical pulse and continuous modulation signals. But the facet parameter structure of the simulated surface can take into account the possibilities of representing various types of terrain and objects created by man. 1 Introduction To develop and improve Earth remote sensing methods, mathematical modeling of the processes of radiation, propagation, reflection, or, in other words, scattering, reception and processing of radar signals is required [1–3]. Modern computing resources make it possible to implement ever more detailed and accurate mathematical models, but in practice they are limited to design specialized models for a particular type of system, type of signal, type of surface, and method for processing and synthesizing radar images [4–6]. The article covers the topic of the implementation of the basic version of a universal mathematical model for a research software package designed to test and evaluate the quality indicators of the algorithms for the operation of radars with synthetic antenna aperture (SAR) in various application conditions. The software package is built from a set of modules combined into a system for modeling reflected signals for a given trajectory of movement, the orientation of the aircraft (as the radar platform) and the characteristics of the underlying surface. * Corresponding author: a.s.bokov@urfu.ru © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). 2 Surface modelling A mathematical model of the surface strongly depends on type of terrain, as well as on possibilities of accounting all the properties of physical process of electromagnetic waves reflection [2, 7]. For the mathematical description of the natural earth's surface, the facet surface model is justified and selected, which is the most universal for various types of * Corresponding author: a.s.bokov@urfu.ru , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 https://doi.org/10.1051/itmconf /20193015026 point, surface-distributed and volume-distributed reflectors of radar targets/terrain [1–5]. It is based on a geometric model of propagation and reflection of radio waves from the entire set of trained reflectors, which are presented in the form of conditional facets, i.e. simplified flat areas with dimensions less than the radar resolution. Each ith facet has a biased backscattering pattern. But the form of a backscattering pattern is equal for the same type of material (substance) of reflecting surface. point, surface-distributed and volume-distributed reflectors of radar targets/terrain [1–5]. It is based on a geometric model of propagation and reflection of radio waves from the entire set of trained reflectors, which are presented in the form of conditional facets, i.e. simplified flat areas with dimensions less than the radar resolution. Each ith facet has a biased backscattering pattern. But the form of a backscattering pattern is equal for the same type of material (substance) of reflecting surface. A typical geometric model of a radio altimeter operation with a radar carrier platform that moves over the facet field, which represents a set of simulated reflectors, is shown in Fig. 1. The typical forms of an altimeter antenna pattern, illuminated footprint and backscattering pattern of a rough surface are also presented there. In the practical case, when the carrier platform moves, the footprint spot also moves with the same speed and direction. Moreover, the radar antenna (and/or its main lobe) may be biased due to the aircraft evolutions or the antenna scanning system. Thus, the simulated surface should be expanded taking into account the entire flight path and angle evolutions of the antenna main lobe. Крупномасштабный рельеф отклоняет ДОР фацетов и влияет на их площадь H Ri Carrier platform Illuminated footprint Nadir point x Antenna pattern y z xmax  Each ith facet has a biased backscattering pattern Facets are the small flat rectangular or triangular areas covering the surface (footprint) Fig. 1. Radar scene with facet field for modeling reflectons from the underlying surface. Carrier platform Each ith facet has a biased backscattering pattern Antenna pattern Facets are the small flat rectangular or triangular areas covering the surface (footprint) Facets are the small flat rectangular or triangular areas covering the surface (footprint) Fig. 1. Radar scene with facet field for modeling reflectons from the underlying surface. * Corresponding author: a.s.bokov@urfu.ru The modeling method used in the work involves the joint use of the phenomenological approach and the totality of empirical data. In the framework of this approach, the facets of the irradiated surface are considered statistically independent with random reflection coefficients, the standard deviation of which depends on the reflector material type based on empirical data on surface elements. Note that, in particular, small inhomogeneities and roughnesses of terrain are excluded from the geometric model of the surface and are taken into account due to the use of diffuse backscattering rather than mirror backscattering. 2.1 Calculation of a set of facet parameters The simulation of the reflected signal begins with the sequential calculation of the slant range, Doppler frequency shift and power for each elementary reflector or surface facet. The slant range is the first base parameter for any radar system that is directly related to the resolution and use of the radar. It is the distance from the radar antenna to the target (or reflector or facet) position. At any time, it can simply be determined from the descriptive geometry of the model shown in Fig. 2. Thus, according with relative coordinates (xi, yi, zi) of the ith facet along each of the 3 axes, the current value of the slant range is determined as:   2 2 2 i i i i R x H y z     (1) (1) 2 2 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 https://doi.org/10.1051/itmconf /20193015026 where H is the radar antenna altitude relative the average surface level. y x z yi βi i  zi xi i Ri 0 ith facet Radar antenna  H Fig. 2. Geometry of the relative positions of the radar antenna and the sample facet. where H is the radar antenna altitude relative the average surface level. where H is the radar antenna altitude relative the average surface level. y Radar antenna Fig. 2. Geometry of the relative positions of the radar antenna and the sample facet. The Doppler frequency shift is the second base parameter for a radar system. Its use is extremely useful to increase radar application. Its definition is a key property for all imaging radars that calculates the images of the underlying surface using special signal processing [1, 7, 8]. Given the geometry of the model shown in Fig. 2, the Doppler frequency shift of the signal reflected from the facet is calculated by the expression: 2υ 2υcos(θ )cos(β ) λ λ i i i if   (2) (2) where i is the radial velocity of the radar platform in the direction of the ith facet; i and i are the angles for taking into account the individual direction of the facet (from the radar antenna, relative to the velocity vector , which is also shown for the ith facet in Fig. 2) in the horizontal and vertical planes. 2.2 Algorithm for modeling the reflected signal As already mentioned, the most common methods of digital modeling of radar signals reflected from complex surfaces and targets are based on facet models. In this case, the entire reflecting surface is divided into some elementary reflecting areas, i.e. facets, and then the signals reflected from each facet as from a point object are calculated. This approach allows one to include various inhomogeneities in the surface that directly affect the nature of the reflected signal. Obviously, it is possible to combine facets with various properties to create complex phono-target environments, and with the dynamics, for example, in a form of a combat area model [7–9]. However, at the first stages of the study of the characteristics of signal processing algorithms in the radar, it is often enough to be limited only to a certain area of space where the concentrated, surface- or volume-distributed target is located. In this case, it is convenient to use a model capable of presenting the background-target environment with sufficient accuracy and with a given detail. The proposed top-level of the algorithm for modeling the raw signal, i.e. signal reflected from the surface, is shown in Fig. 3. Calculation of the size and position of the center of the illuminated spot Input data entry Formation of facet fields, their coordinates and parameters Calculation of signal power reflected from one facet Building a signal model reflected from one facet Is it the last time step? Change the current coordinates of the aircraft in accordance with  Change the current value of the time variable yes no Is it the last facet? Summing a signal from the current facet with a common signal Resetting the current value of a time variable yes Change the current coordinates of the aircraft in accordance with  Raw signal output no Fig. 3. Algorithm for modeling the raw signal reflected from the surface. Is it the last facet? Change the current coordinates of the aircraft in accordance with  Fig. 3. Algorithm for modeling the raw signal reflected from the surface. 2.1 Calculation of a set of facet parameters where i is the radial velocity of the radar platform in the direction of the ith facet; i and i are the angles for taking into account the individual direction of the facet (from the radar antenna, relative to the velocity vector , which is also shown for the ith facet in Fig. 2) in the horizontal and vertical planes. The power of the signal reflected from the target is the third base parameter for a radar system. In the case of terrain modeling using the facet model, any facet is a small target. Thus, the reflected facet power is in accordance with the radar equation, which can be written as the following expression:    2 2 0 3 4 λ η (4π) ant i surf i i i i i G K G B S P P R  (3) (3) where P0 is the radar transmitter power;  is the carrier wavelength; Gant is the antenna gain; Ki is the coefficient of accounting of the antenna pattern; Gsurf is the reflection coefficient to account for the specific surface or target radar cross section (RCS); Bi is the coefficient of the backscattering pattern; Si is the facet area; i is the coefficient of accounting for losses in the distribution medium (for example, including shading of the rain drops or vegetation layer, if present). To summarize, we can say that the structure of the intrinsic and calculated parameters of the facet of the universal mathematical model should include the above ()i parameters in order to take into account the possibilities of representing various types of landscape and artificial objects. Some additional parameters for representing natural surfaces will be discussed below. 3 3 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 https://doi.org/10.1051/itmconf /20193015026 2.2 Algorithm for modeling the reflected signal Also, for adequate signal modeling, it is necessary to take into account the initial data of the trajectory of the aircraft, radar, such as the altitude and components of the aircraft’s flight speed vector, the angles of evolution of the aircraft (roll and pitch), the parameters of the radiation patterns of the receiving and transmitting elements of the antenna system (the width and shape of the radiation patterns), angles of deviation of the antenna axes from the vertical axis of the aircraft, the carrier frequency of the transmitter, the length of the probe pulse, the duty cycle of the pulses and the number of pulses emitted on the simulation path (for a pulse radar ), frequency deviation, modulation period and the number of modulation periods on the modeling path (for continuous-wave radars), signal sampling frequency. 4 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 https://doi.org/10.1051/itmconf /20193015026 3.1 Homogeneous rough surface model Due to the fact that in the general case, for a homogeneous rough surface, the position of all reflectors is random, such a model makes it possible to take into account any (typical) radar situation in a simplified form. Its use is effective in the statistical study of reflected signals with an unknown (arbitrary) type of surface. The coordinates of the facet arrangement in the plane are random variables uniformly distributed within the irradiation spot. The vertical coordinate has a normal distribution, the dispersion of this value is set in advance and is an indicator of large surface roughness. An example of such a facet field for nadir irradiation for a height of 100 m, the size of large relief irregularities is 2.5 m, the number of facets is 1000, and with an irradiation spot radius of 50 m, is shown in Fig. 4. -40 -20 0 20 40 -40 -20 0 20 40 -5 0 5 ( -40 -20 0 20 40 -40 -20 0 20 40 -5 0 5 (x, y, z), m Fig. 4. Facet field of modeling reflectors of a homogeneous rough underlying surface. (x, y, z), m Fig. 4. Facet field of modeling reflectors of a homogeneous rough underlying surface. 3 Some exaples of mathematical modeling The objects of research were the following types of terrain: 1) a homogeneous rough surface, 2) a sea surface, 3) a forest surface. For an accurate calculation of the energy distribution in the spectrum, a three- dimensional mathematical model of the “radar – underlying surface – radar” channel is necessary, taking into account both the principles of radar signal processing, as well as the parameters of the underlying surface, velocity and evolution of the aircraft [8, 9]. For the greatest versatility, modeling of reflective underlying surfaces is carried out by creating a facet field consisting of elementary point reflectors. The facet coordinates can be set in various ways: randomly, in a square or rectangular lattice, in a hexagonal or triangular lattice, along concentric circles, in a spiral, etc. The number of faces must match the radar resolution for the range and angular coordinates. 3.2 Sea model When studying the sea surface, it is necessary to take into account factors influencing the wave formation of the sea, such as: speed and direction of the wind, the length of the region of acceleration of waves, ocean movements, etc. The sea surface modeling algorithm can account the model of the energy spectrum of sea waves (capillary for ripples, TMA for wind waves), the parameters for each elementary wave in the wave composition (height and wavelength, direction of propagation, wave phase, etc.) which are determined from this spectrum [6]. But in simple case (howewer, useful to study) it can be a one harmonic wave which moves in the desired direction. Examples of a complex model and such a simplified facet field for a sea wave height of 7 m, the speed of the wave along the Z axis is 7 m/s, along the Y axis is 4 m/s, the number of facets is 2000, and with an irradiation spot radius of 50 m, is shown in Fig. 5. 5 5 5 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 , (2019) ITM Web of Conferences 30 15026 https://doi.org/10.1051/itmconf /20193015026 CriMiCo'2019 -50 0 50 -50 0 50 (x, y, z), m Fig. 5. Complex model and simplified facet field for modeling sea surface reflectors. Fig. 5. Complex model and simplified facet field for modeling sea surface reflectors. 4 Conclusion In this paper the algorithm that uses a phenomenological model to describe underlying surfaces was developed. The universality of the considered model of radar signals lies in the possibility of reducing the amount of calculations by using the model of point reflectors while maintaining the main features of the reflected signal formation as the sum of the reflected signals, taking into account the amplitude and phase ratios of the partial signals. Such a simplification is correct from the point of view of developing and debugging algorithms for joint work of the formation and processing models (determining the relative range/altitude, synthesizing radar images) of the signals reflected from various types of natural and humanmade objects when modeling and studying the characteristics, resolution, relative power budgets of various radar and altimeter systems, including SAR. It is worthwhile to note, that mathematical modeling of radar system operation is the main way to study common and state-of-the-art methods of radar signal processing, as well as to develop new, more efficient, reliable and accuracy methods of radiovision of various vehicles. Acknowledgments This research has been funded by a grant of the Ministry of Science and Higher Education of the Russian Federation (project No. 8.2538.2017/4.6). 3.3 Forest model 6 6 https://doi.org/10.1051/itmconf /20193015026 , (2019) ITM Web of Conferences 30 CriMiCo'2019 15026 3.3 Forest model It was experimentally established that in the centimetre wave band the signal from the canopy is comparable in power with the signal from the underlying soil (ground) [4–6, 9], and, with a resolution of 1 m or less, it is well localized, both in the presence of foliage and in the absence of it. Although, in a number of works, the forest surface is assigned to surface-distributed targets [2, 3], it is more correct to consider it layered or volume- distributed, because forest canopy as a radar target has a distribution both on the surface and in depth. The signal reflected from it is more “blurred” in range than the signal reflected from other types of underlying surfaces. Forest surface is a complex heterogeneous surface that can be divided with some simplification into three layers: soil, undergrowth, trees (trunk, branches, leaves). During the modeling process the first stage, the soil layer is set, which is a plane with a small topography, the boundaries and area of which depend on the aircraft height and antenna pattern for a given aircraft motion path. Then, the layered structure for the undergrowth and tree canopy is simulated. Examples of a complex model [9] and such a simplified forest facet field for the first tree level with height of trees of about of 30 m (number of facets is 220), with the small tree’s height of 10 m (number of facets is 100), with the number of facets is 1000 for the ground level with the size of large relief irregularities is 2.5 m,, is shown in Fig. 6. -50 0 50 -50 0 50 0 10 20 30 (x, y, z), m Fig. 6. Complex model and simplified facet field for modeling forest reflectors. -50 0 50 -50 0 50 0 10 20 30 (x, y, z), m (x, y, z), m Fig. 6. Complex model and simplified facet field for modeling forest reflectors. Unfortunately, it is difficult to implement fast and complete complex models for sea and forest surfaces. The choice of a model is largely determined by the availability of sufficient computing resources, that is why we are currently concentrating on some simplifications [9]. However, the proposed simplified facet models produce the desired level of quality for generated raw signals [10], because these signals include all the basic surface parameters that are needed for various radar systems, including SAR. References 1. B. R. Mahafza, Radar Systems Analysis and Design Using MATLAB (Chapman and Hall/CRC, 2013) 2. A. I. Baskakov, T. S. Zhutyayeva, Yu. I. Lukashenko, Locational Methods for Studying Objects and Media (in Russian, Academy, Moscow, 2011) 3. S. G. Zubkovich, Statistical characteristics of radio signals reflected from the Earth's surface (in Russian, Soviet radio, Moscow, 1968) 4. P. Liang, M. Moghaddam, L. E. Pierce, R. M. Lucas, IEEE Transactions on geoscience and remote sensing, 43, no. 11, 2612–2626 (2005). 5. F. T. Ulaby, M. C. Dobson, Handbook of radar scattering statistic for terrain (Artech house, USA, 1989) 6. A. K. Fung, K. S. Chen, Microwave Scattering and Emission Models for Users (Artech house, USA, 2010) 7. V. G. Kobernichenko, Radioelectronic Earth`s remote sensing systems (in Russian, UrFU, Yekaterinburg, 2016) 8. G. S. Kondratenkov, А. U. Frolov, Radiovision. Radar Earth`s remote sensing systems (in Russian, Radiotechnica, Moscow, 2005) 9. A. S. Bokov, A. E. Smertin, V. G. Vazhenin, CEUR-WS, 2274, 42–51 (2018). 10. Yu. V. Markov, A. S. Bokov, V. G. Vazhenin, S. A. Margilevsky, RSEMW-2019, 352–355 (2019) 7
https://openalex.org/W4387704720	https://www.cambridge.org/core/services/aop-cambridge-core/content/view/3C143E501E0824C8F9F0C40925965F43/S1537592723002827a.pdf/div-class-title-a-global-analysis-of-transgender-rights-introducing-the-trans-rights-indicator-project-trip-div.pdf	English	null	A Global Analysis of Transgender Rights: Introducing the Trans Rights Indicator Project (TRIP)	Perspectives on politics	2,023	cc-by	14,649	A Global Analysis of Transgender Rights: Introducing the Trans Rights Indicator Project (TRIP) Myles Williamson To what extent do countries protect the rights of transgender people? How does this diﬀer from legal protections countries oﬀer sexual orientation minorities? What conditions are beneﬁcial for advancing trans rights? Limitations in data availability and accessibility make answering these types of trans-speciﬁc questions diﬃcult. To address this shortcoming, I introduce a new dataset. The Trans Rights Indicator Project (TRIP) provides insight into the legal situations transgender people faced in 173 countries from 2000 to 2021. The dataset currently includes 14 indicators that capture the presence or absence of laws related to criminalization, legal gender recognition, and anti-discrimination protections. I then use this data to discuss the global status of transgender rights throughout the period and compare these trends to sexual orientation rights. Finally, I conclude with a preliminary analysis of three institutional and cultural factors that may help explain variation in transgender rights throughout the world. A et al. 2014; Longaker and Haider-Markel 2014). Doing so neglects the rights unique to transgender individuals and makes important questions about this group challenging to answer. A reignited wave of backlash continues to threaten the rights of transgender people throughout the world. In the United States, legislatures across the country have put forward hundreds of anti-trans bills, targeting rights such as access to gender-aﬃrming health- care and legal gender recognition (ACLU 2023). Similarly, countries like Hungary and Russia have adopted and expanded anti-LGBT1 propaganda laws to further restrict freedoms of expression for gender identity minorities (Reuters 2021, 2022). Despite the direct attacks that transgender people face, political science scholarship rarely examines these individuals and their rights on a global scale. Instead, studies tend to treat the LGBT community as a homogenous group, often using measures related to sexual orientation as a proxy for transgender rights (Lewis The tendency to conﬂate sexual orientation and gender identity in cross-national LGBT research arises from limitations in data availability and accessibility (Haider-Markel et al. 2019). To address this shortcom- ing, I introduce a new dataset that allows for more nuanced research on transgender rights separately from the broader LGBT community. The Trans Rights Indicator Project (TRIP) provides insight into the legal situations transgender people faced in 173 countries from 2000 to 2021. The dataset currently includes 14 indicators that capture the presence or absence of laws related to criminalization, legal gender recognition, and anti-discrimination protections. Article Article doi:10.1017/S1537592723002827 © The Author(s), 2023. Published by Cambridge University Press on behalf of the American Political Science Association. This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited. September 2024 \| Vol. 22/No. 3 799 A Global Analysis of Transgender Rights: Introducing the Trans Rights Indicator Project (TRIP) I draw from a variety of nongovernmental and international organizations, national law documents, research institutes, and other sources to code each variable. A list of permanent links to Supplemental Materials provided by the authors precedes the References section. Data replication sets are available in Harvard Dataverse at: https://doi.org/10.7910/DVN/FXXLTS Data replication sets are available in Harvard Dataverse at: https://doi.org/10.7910/DVN/FXXLTS In addition to introducing the new dataset, this article contributes to the literature on LGBT politics and human rights by addressing three questions. First, to what extent do countries protect the rights of transgender people, and how has this changed over time? I ﬁnd that while many countries have progressed over the last two decades, legal protections for transgender individuals remain limited throughout most of the world. This descriptive exercise Myles Williamson is a Doctoral Candidate in the Department of Political Science at the University of Alabama (eswilliamson@crimson.ua.edu). His research focuses on the intersection of LGBT politics and human rights in diﬀerent political regimes. He is the principal investigator for the Trans Rights Indicator Project (TRIP). doi:10.1017/S1537592723002827 doi:10.1017/S1537592723002827 doi:10.1017/S1537592723002827 © The Author(s), 2023. Published by Cambridge University Press on behalf of the American Political Science Association. This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0), which permits unrestricted re-use, distribution and reproduction, provided the original article is properly cited. September 2024 \| Vol. 22/No. 3 799 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Article \| A Global Analysis of Transgender Rights identity and assigned sex diﬀer, and “cisgender” for those whose gender identity and assigned sex correspond.5 not only provides important context on the global state of trans rights, but also enables policymakers and advocacy organizations to identify the areas where these minorities may be most vulnerable to abuse from the state and society.2 Importantly, notions of gender identity and the words to describe it remain ever-evolving and may vary consid- erably across cultures. Recognizing this diversity, trans- gender in this context is not a deﬁnitive label indicative of one speciﬁc gender identity. Instead, the term serves as a descriptor for individuals with any gender identity that diverges from their birth-assigned sex. Thus, this article deﬁnes transgender in a way that is narrow enough to adequately capture the intended focal point while still being broad enough to apply in a cross-national context. Though this terminology reﬂects just one possible under- standing of gender identity, it still provides an important lens for studying gender globally. y Second, how do the legal rights that countries provide sexual orientation and gender identity minorities diﬀer from one another? Comparisons between the TRIP indi- cators and data on LGB rights demonstrate that countries do not inherently treat sexual- and gender identity minor- ities equally. Therefore, conﬂating measures of the two groups may skew our understanding of the actual rights that countries provide to either. Even when countries appear to oﬀer similar levels of legal rights across the LGBT community, the comparisons emphasize the dif- ferences in policy concerns between the groups. The rights speciﬁc to gender identity minorities deserve scholarly attention separately from the broader community. 800 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press doi:10.1017/S1537592723002827 Finally, sexual orientation refers to an individual’s “inherent or immutable enduring emotional, romantic, or sexual attraction to other people” of the same or diﬀerent genders (Human Rights Campaign 2021).6 While the language that describes someone’s sexual orien- tation may shift depending on a person’s gender identiﬁ- cation, sexual orientation does not dictate gender identity and vice versa (Stryker 2008). For example, a person who identiﬁes as a man and is attracted to men shares the same gender identity as someone who identiﬁes as a man but is attracted to women, despite having diﬀerent sexual orientations. Finally, what conditions are favorable for advancing transgender rights? In short, I ﬁnd that democracy and economic development are positively associated with trans rights, whereas the percentage of religious adherents in society appears to be insigniﬁcant. Though additional research is necessary to further investigate these relation- ships, this preliminary analysis provides an initial look into what makes countries more or less likely to protect the rights of transgender people—a task that becomes increas- ingly important as attacks on gender diversity spread across the globe. Differentiating Gender Identity and Sexual Orientation For one, data collection requires ﬁrst knowing what rights are relevant to the trans community. Transgender people constitute a highly marginalized and relatively small por- tion of the population. This limits the visibility of these individuals and their legal needs, making it more challeng- ing to identify which rights are important for data collec- tion. Additionally, this marginalization limits the reliability of data on de facto measures of rights. For example, reports of violence or discrimination against trans individuals likely undercount the actual rights vio- lations that occur, especially in countries that oﬀer no other protections to trans people. compares political power distribution speciﬁcally based on sexual orientation not (trans)gender identity (Coppedge et al. 2022). No measures appear to represent speciﬁc dimensions or indicators of transgender rights. Alternatively, F&M Global Barometers oﬀer several trans-speciﬁc measures with their “Global Barometer of Transgender Rights” (Dicklitch-Nelson and Rahman 2022). The GBTR includes 12 variables explicitly mea- suring trans rights and ﬁve variables capturing LGBT advocacy protections for 204 countries.9 However, the data currently only spans from 2011 to 2019, and the raw data ﬁles are not (yet) accessible. Additionally, ﬁve of the twelve trans-speciﬁc variables are de facto measures of societal persecution. These measures can be beneﬁcial for gaining insight into the realities within a country but are less reliable than de jure indicators due to possible underreporting of these rights violations. Even when only focusing on legal rights, collecting this type of data on a global scale over several years is an intensive and time-consuming process. Transgender rights span across a wide variety of legal areas, which requires searching through a myriad of laws, policies, and other sources to determine if a country provides a speciﬁc right. This process also requires locating and interpreting addi- tional information about the legal situation that may not be apparent by the law alone. For example, in the United States, Title VII of the Civil Rights Act of 1964 prohibits employment discrimination based on sex (among other factors) but does not explicitly mention (trans)gender identity or expression. However, in 2020, the Supreme Court’s decision in Bostock v. Clayton County held that “it is impossible to discriminate against a person for being homosexual or transgender without discriminating against that individual based on sex.”7 Therefore, the binding decision extends the legal right to transgender people, even though the original text of the law does not explicitly do so. Differentiating Gender Identity and Sexual Orientation g While existing scholarship provides valuable insight into our understanding of LGBT politics more broadly, few studies address transgender rights speciﬁcally. Instead, most of what we know about trans rights builds on the assumption that they resemble lesbian, gay, and bisexual rights (Haider-Markel et al. 2019; Tadlock and Taylor 2017). Despite conceptual diﬀerences, research often conﬂates sexual orientation and gender identity under the umbrella acronym without adequately examining the latter (Paternotte 2018). Meanwhile, political science studies of gender diverse minorities are often limited to single or regional case studies, largely focusing on Western countries (Dicklitch-Nelson and Rahman 2022; Haider- Markel et al. 2019). Though often discussed collectively, sex, gender identity, and sexual orientation are distinct concepts referring to diﬀerent aspects of a person’s being (Meyerowitz 2002). Given their conceptual complexity, this section provides a deﬁnition of each term as it is utilized within the context of this article. To start, sex refers to an individual’s “repro- ductive capacity or potential” as exhibited by the physical and chromosomal characteristics of the body (Stryker 2008, 8). The determination of sex typically occurs at birth based on observable features related to the reproduc- tive system. Contemporary sex classiﬁcations often include categories such as male, female, and intersex.3 g While sex concerns the physical body, gender identity refers to an individual’s “innermost concept of self” regarding being male, female, both, or neither (Human Rights Campaign 2021).4 In other words, a person’s gender identity encapsulates their internal sense of self regardless of physical or sex-based characteristics (Stryker 2008). Given this distinction, an individual may possess a gender identity that diverges from their birth-assigned sex. For example, a person’s gender identity may be male despite being assigned female at birth. This article uses “transgender” (trans) to describe individuals whose gender Though sexual orientation and gender identity minor- ities may coalesce around the LGBT identity and share similar experiences, policy concerns speciﬁc to transgender people can diﬀer signiﬁcantly from those of the LGB community (Schwenke 2021; Thiel 2014). For example, laws allowing gender marker changes on identity docu- ments may be important for some trans individuals. However, these laws are likely irrelevant for cisgender LGB people. Thus, “LGBT” research only using measures related to sexual orientation rights misses important trans- speciﬁc policies. The underrepresentation of transgender rights in cross- national research stems from limitations in available data. Several factors make collecting this type of data diﬃcult. Differentiating Gender Identity and Sexual Orientation Finally, ILGA World and its regional branch, ILGA Europe, provide perhaps the most well-known data on transgender rights. ILGA World has published three editions of its Trans Legal Mapping Report since 2016 (Chiam, Duﬀy, and Gonzales Gil 2016, 2017; Chiam et al. 2020). The ﬁrst two editions provide data on the legality of name and gender marker changes, while the third edition also includes information on criminalization in select countries. ILGA Europe’s “Rainbow Europe” map documents an extensive list of various sexual orientation and gender identity rights.10 However, the spatial dimension is limited to European countries. While citing some ILGA data, TRIP also includes indicators, countries, and years not covered by these sources and provides the data in a country-year format to allow for quantitative assessments. In sum, diﬀerences among the LGBT community warrant disaggregated data. Even if countries treat these groups similarly, transgender people are a separate minor- ity with their own policy concerns that deserve individu- alized consideration. While a few studies incorporate related measures, work that addresses trans rights in greater depth independently of the broader LGBT community remains limited. To help ﬁll this gap, I present a new dataset tracking transgender rights worldwide. The fol- lowing section introduces this dataset and details the data collection and coding processes. Despite data challenges, a few notable empirical studies include some cross-national considerations of transgender rights.8 For example, Badgett et al. (2014) supplement their LGB index with data from Transgender Europe. They include 16 variables related to legal document changes, legal protection, criminalization or state- sponsored discrimination, and healthcare. Unfortunately, they only have trans-speciﬁc data for 2012 and only for 18 of the 39 countries in their analysis. Lee and Ostergard (2017) also incorporate some measures of trans rights in their LGBT discrimination index. However, this is pri- marily limited to variables capturing intolerance towards the broader “LGBT” community, making separate ana- lyses of trans rights impossible. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press The Trans Rights Indicator Project Methodology The Trans Rights Indicator Project (TRIP) provides country-year data on legal rights protections relevant to transgender minorities (Williamson 2023). The data includes 14 trans-speciﬁc variables for 173 countries from 2000 to 2021. TRIP is among the ﬁrst to explicitly focus on trans rights and the only public dataset covering trans rights with wide spatial and temporal coverage (to my knowledge). The dataset excludes de facto societal Varieties of Democracy (V-Dem) has a measure that contrasts the distribution of political power between “het- erosexuals and lesbian, gay, bisexual, and transgender (LGBT) members of the polity” (Coppedge et al. 2022, 208). Like Lee and Ostergard (2017) though, the measure does not have separate values for sexual orientation versus gender identity minorities. Additionally, the variable September 2024 \| Vol. 22/No. 3 801 Article \| A Global Analysis of Transgender Rights measures, given information availability and accuracy concerns discussed in the previous section. While laws may not always represent lived experience, the legal envi- ronment still provides valuable insight into the climate towards transgender people in a country (Htun and Weldon 2012; Velasco 2018). Direct criminalization refers to laws that explicitly criminalize individuals based on gender identity or expres- sion when it diverges from the traditional expectations of birth-assigned sex. These laws typically include provisions against “cross-dressing” or “disguising” as a diﬀerent sex/ gender.11 This indicator equals one if criminalizing laws exist at the subnational or national level and equals zero otherwise. Indirect criminalization occurs when countries arbitrarily use laws unrelated to gender identity or expres- sion to target transgender people. For example, security forces may use laws concerning “public order,” “vagrancy,” “morality,” “decency,” and other vague oﬀenses to indirectly criminalize these individuals (Chiam et al. 2020; Egerton-Peters et al. 2018). Laws only qualify as indirect criminalization if at least one credible source states that countries used them to target transgender people. I also include laws policing consensual same-sex sexual activity as indirect criminalization due to the potential conﬂation of sexual and gender identities. Developing and Measuring Indicators of Trans Rights This project rests on the idea that transgender rights are those that allow trans individuals to exist and participate in government and society as their gender in the same manner guaranteed to all other individuals. To operatio- nalize these rights, TRIP builds from ideals set forth by international human rights law, namely the Yogyakarta Principles. The Trans Rights Indicator Project Methodology Table 1 TRIP indicators Criminalization (national and subnational) Direct criminalization Indirect criminalization Legal Recognition (national) Legal gender marker change (GMC) Presence of prohibitive requirements for GMC - Physiological requirements - Psychological requirements - Divorce/single-status requirements Nonbinary/third gender marker options Legal Protections (national) General (non-sector speciﬁc) anti-discrimination protections Constitutional anti-discrimination protections Employment anti-discrimination protections Education anti-discrimination protections Healthcare anti-discrimination protections Housing anti-discrimination protections Table 1 TRIP indicators Criminalization (national and subnational) Direct criminalization Indirect criminalization Legal Recognition (national) Legal gender marker change (GMC) Presence of prohibitive requirements for GMC - Physiological requirements - Psychological requirements - Divorce/single-status requirements Nonbinary/third gender marker options Legal Protections (national) General (non-sector speciﬁc) anti-discrimination protections Constitutional anti-discrimination protections Employment anti-discrimination protections Education anti-discrimination protections Healthcare anti-discrimination protections Housing anti-discrimination protections Table 1 TRIP indicators Criminalization (national and subnational) Direct criminalization Indirect criminalization Legal Recognition (national) Legal gender marker change (GMC) Presence of prohibitive requirements for GMC - Physiological requirements - Psychological requirements - Divorce/single-status requirements Nonbinary/third gender marker options Legal Protections (national) General (non-sector speciﬁc) anti-discrimination protections Constitutional anti-discrimination protections Employment anti-discrimination protections Education anti-discrimination protections Healthcare anti-discrimination protections Housing anti-discrimination protections The Trans Rights Indicator Project Methodology These principles outline countries’ obligations to protect and promote the well-being of sexual orientation and gender identity minorities (ICJ 2007, 2017). Drawing from these principles, I organize indica- tors into three categories: criminalization, recognition, and protection (table 1). Indirect criminalization is “present” for all years the law is in force. In a few instances, sources report arbitrary arrests and criminalization but do not cite speciﬁc laws or charges. In these cases, criminalization is only “present” for the year the incidents occur, according to the sources. This indicator equals one when indirect criminalization is “present” at the subnational or national level. The indica- tor may undercount instances of this type of criminaliza- tion, particularly in countries with no LGBT advocacy organizations that track these incidents. However, I still include this measure due to the widespread occurrence of indirect criminalization. Criminalization threatens the realization of all other rights related to an individual. Yogyakarta Principle 33 states that everyone has “the right to be free from criminalization and any form of sanction arising directly or indirectly from that person’s actual or perceived sexual orientation, gender identity, gender expression or sex characteristics” (ICJ 2017, 11). For transgender people, the threat of criminalization is two-fold. These individuals are vulnerable to laws that directly police gender identity/ expression and unrelated laws like those concerning sexu- ality or morality (Egerton-Peters et al. 2018). Therefore, TRIP includes two separate indicators to measure direct and indirect criminalization. The second category of indicators concerns the legal recognition of transgender individuals. Yogyakarta Prin- ciples 3, 31, and 32 espouse individuals’ rights to legal recognition based on self-determined gender identity (ICJ 2007; 2017). Oﬃcial identity documents serve as critical tools that often determine individuals’ legal and social status (Holzer 2022). Having a legal gender marker that is incongruent with someone’s identity or expression may make that person more susceptible to discrimination or other abuses. Table 1 TRIP indicators For example, 32% of trans respondents in the United States and an average of 25% across the European Union experienced discrimination after showing an identity doc- ument not matching their gender presentation (James et al. 2016).12 Legal gender incongruencies can also make trans individuals more vulnerable to arrest and discrimi- nation by government oﬃcials, as evidenced by the cases of Panama and Peru following the COVID-19 sex-based lockdowns (Edgell et al. 2021). Police oﬃcers arrested trans individuals for appearing in public on the “wrong” day when their perceived identity did not match their legal gender marker (Cabrera 2022). To capture legal recognition, TRIP contains indicators covering the legal right to gender marker changes, 802 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press prohibitive requirements for those changes, and oﬃcial recognition of nonbinary/third genders. The ﬁrst indicator measures whether a nationally applicable law or policy allows gender marker changes on identity documents beyond travel documents alone. The indicator equals one if national law allows these changes. Countries receive a zero if national law does not specify this right (e.g., the United States) or if gender marker changes are prohibited (e.g., Hungary). For this variable, I do not count subna- tional laws or established practices that countries have not codiﬁed into national law. 2 demands that countries protect the right of equality and non-discrimination for individuals of all sexual orientations and gender identities (ICJ 2007). Anti- discrimination protections are critical for protecting mar- ginalized communities and allowing these individuals to participate more fully in government and society (Fields and Wotipka 2022). To measure the extent of legal pro- tections a country oﬀers, TRIP includes six indicators for the following types of national anti-discrimination laws: general (non-sector speciﬁc), constitutional, employment, education, healthcare, and housing.13 When countries allow gender marker changes, they often impose prohibitive requirements that individuals must meet to amend their documents. I include one broad indicator that captures whether the country has any prohibitive requirements and three speciﬁc indicators based on the type. These include physiological, psycho- logical, and divorce/single-status prerequisites. Physiolog- ical requirements refer to any medical interventions that an individual must undergo to alter their body or sex charac- teristics. This may include physical interventions such as hormone therapy, surgery, or sterilization. Psychological prerequisites stipulate that individuals receive a diagnosis or documented treatment by a psychiatrist, psychologist, or other mental health medical professional. Table 1 TRIP indicators Finally, divorce/single-status provisions require individuals to be divorced or otherwise unmarried to qualify for a legal gender marker change. This enables countries to allow legal gender marker changes without recognizing same-sex marriages. General anti-discrimination laws and policies are those that are not entirely speciﬁc to one sector. While they might also contain sector-speciﬁc provisions, these laws provide broad-based protections from discrimination against certain characteristics of individuals or groups. For example, Montenegro’s 2010 Law on Prohibition of Discrimination prohibits “any form of discrimination, on any ground” and explicitly states that “any diﬀerentiation, unequal treatment or bringing a person in an unequal position based on gender identity or sexual orientation, shall be deemed to be discrimination.”14 Constitutional protections refer to anti-discrimination provisions that are speciﬁcally embedded within a coun- try’s constitution. For instance, Article 14 of Bolivia’s 2009 constitution explicitly prohibits discrimination based on several grounds, including gender identity and sexual orientation.15 Though constitutional protections do not guarantee that countries will adopt other anti- discrimination policies, these provisions help establish a more inclusive legal foundation. Lastly, indicators for employment, education, healthcare, and housing capture the presence of anti-discrimination laws and policies rel- evant to those areas. These types of laws can vary in the exact protections provided from country to country, but at their core must include some provision protecting trans- gender individuals from discrimination or unfair treat- ment within those respective sectors. Indicators for each prohibitive requirement equals one if present at the national level and zero if not. The broad indicator equals one if a country has any physiological, psychological, or divorce/single-status requirements in place. Importantly, a value of zero for each prohibitive requirement in the TRIP data does not guarantee a country has a self-determination model for gender recog- nition. Countries may have other requirements beyond those captured in the data (e.g., judicial procedures). Indicators for each of the anti-discrimination protec- tions equal one only when national laws or policies exist. Additionally, the provisions must explicitly protect indi- viduals on the grounds of gender identity, gender expres- sion, or gender “reassignment.”16 Implicit inclusion on the grounds of sex does not count unless a binding and nationally applicable court ruling or policy speciﬁes that “sex” includes gender identity, expression, or “reassignment.” p g j p The last indicator in this category measures whether countries have national recognition of nonbinary or third gender markers on identity documents. Table 1 TRIP indicators I use the terms “nonbinary” or “third gender” to mean any gender desig- nation outside of the traditionally issued binary markers in each country (e.g., “male” or “female”). Examples include the “Other” (“Anya”) designation in Nepal and the “X” marker in Malta. In some cases, these markers are available but reserved for individuals with speciﬁc identities, as in the case of the “Hijra” designation in Bangladesh. These individuals may or may not self-identify as transgender, but I include these cases since an alternate marker is available. This indicator equals one if countries recognize nonbinary/third gender markers and zero if not. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press TRIP Scores As the earlier example of the United States’ employment discrim- ination law demonstrates, additional policies or court decisions may extend a speciﬁc right to transgender people despite the text of the original legal provision suggesting otherwise. I also deferred to secondary sources in cases where the full text of the law was inaccessible or unavail- able in an English translation. 13 table 2, I calculate the composite score by summing the number of indicators a country performs well on each year (Dicklitch-Nelson and Rahman 2022; Velasco 2018). For example, a raw score of four means that values equal one on four indicators for a given country that year. Since higher scores represent greater rights protections, I inverted negative indicators related to criminalization and prohibitive requirements for gender recognition. Positive-framed indicators remained the same since their values already equal one when the country provides the respective right. One concern when using a wide variety of secondary sources is the consistency of the level of measurement, as sources have diﬀerent parameters for counting a right as “present.” For example, some sources count sub-national and national laws, while others may only discuss measures at the national level. This was even more complicated when sources did not clearly state this information (e.g., U.S. Department of State Human Rights Reports). Due to these potential concerns, I used two or more sources to code each variable, when possible, to make sure the data accurately reﬂects the parameters set out in the TRIP codebook. The TRIP score is useful for gaining a broad under- standing of how trans-friendly a country’s laws may be at the national level and for comparing these rights with other measures of human rights. However, the richness of the data lies within the individual indicators. Since coun- tries can theoretically arrive at similar scores through diﬀerent means, each of the indicators oﬀers a more detailed look into the diﬀerent domestic legal situations. The structure of the TRIP dataset allows for researchers to analyze the indicators individually or reweighted in various manners. Given the scope of this dataset, I used multiple words and phrases to locate and search through electronic sources. The term “transgender” is not universal across time and space (Beemyn 2014; Thiel 2014). TRIP Scores In addition to the individual indicators, TRIP presents a yearly score for each country to represent the total domestic legal rights available for transgender people. Possible scores range from 0 to 13, with higher scores indicating more rights. Using the indicators listed in The ﬁnal category of rights contains variables concern- ing legal discrimination protections. Yogyakarta Principle September 2024 \| Vol. 22/No. 3 803 Article \| A Global Analysis of Transgender Rights organizations such as Amnesty International, Human Dignity Trust, Human Rights Watch, ILGA World and its regional branches, and Transgender Europe. I also consulted the list of member organizations provided by ILGA World to gather information from domestic LGBT or trans-speciﬁc organizations for each country. Table 2 TRIP indicators for score calculations Indicators Score No direct criminalization 1 No indirect criminalization 1 Legal gender marker changes (GMC) possible 1 No physiological requirements for GMC 1 No psychological requirements for GMC 1 No divorce/single-status requirements for GMC 1 Nonbinary/third gender marker options available 1 General anti-discrimination protections present 1 Constitutional anti-discrimination protections present 1 Employment anti-discrimination protections present 1 Education anti-discrimination protections present 1 Healthcare anti-discrimination protections present 1 Housing anti-discrimination protections present 1 Total 13 Table 2 TRIP indicators for score calculations Indicators Score No direct criminalization 1 No indirect criminalization 1 Legal gender marker changes (GMC) possible 1 No physiological requirements for GMC 1 No psychological requirements for GMC 1 No divorce/single-status requirements for GMC 1 Nonbinary/third gender marker options available 1 General anti-discrimination protections present 1 Constitutional anti-discrimination protections present 1 Employment anti-discrimination protections present 1 Education anti-discrimination protections present 1 Healthcare anti-discrimination protections present 1 Housing anti-discrimination protections present 1 Total 13 Beyond NGOs, I searched through reports from inter- national organizations such as the Council of Europe, the European Union, the Organization for Economic Co-operation and Development, and the United Nations. In addition to these sources, I accessed individual govern- ment websites to locate relevant laws and reports. I also reviewed relevant human rights reports from the U.S. Department of State and the Williams Institute. Finally, I consulted news articles to supplement the infor- mation from other sources as needed. The relevant laws and policies served as the ultimate deciding factor for coding each indicator. However, I utilized secondary sources to help identify the presence or absence of any legal provisions and to gain additional context on the applicability of those provisions. TRIP Scores For instance, sources commonly used the word “transexual” in earlier years covered by the data.18 I also considered the possibil- ity of sources using words with diﬀerent connotations to describe individuals who do not identify with their birth- assigned sex. A source may use words like “transvestite” or “cross-dresser,” though it is describing a person this project refers to as “transgender.” In these cases, I relied on the surrounding context provided by the source to determine its relevancy. Regional and cultural diﬀerences further contribute to the variety of words used to describe these individuals. The complete list of search terms I used 804 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Collecting the Data h b For each observation, I hand-coded information from multiple organizations, government websites, national law documents, research institutes, and news sources.17 I began with reports published by nongovernmental related to “transgender” can be found in the codebook for the dataset. considerable improvements. Conversely, indirect crimi- nalization, nonbinary recognition, and constitutional pro- tections constitute the smallest positive changes. Direct criminalization is the only indicator in which countries performed worse in the ﬁnal year. The following sub- sections further expand on these changes to provide more context on the global status of transgender rights. The Global Status of Transgender Rights, 2000–2021 Based on the TRIP data, transgender rights at the turn of the century were scarce. In 2000, the mean TRIP score across the globe was approximately 11%, and no country performed well on more than four indicators. The most protective countries were those where both forms of criminalization were absent and gender marker changes were possible. However, all countries with legal gender recognition only allowed binary marker changes, which were contingent on individuals fulﬁlling at least one type of prohibitive requirement. Additionally, trans-inclusive anti-discrimination protections were nonexistent globally. While still largely hostile, the legal situation improved in many countries by the ﬁnal year in the data. The average global TRIP score roughly doubled by 2021 and mean scores for every region except for the Middle East and North Africa also increased over the period (ﬁgure 1). Criminalization l Direct criminalization of transgender people increased over the last two decades but remains relatively uncom- mon. In 2000, eight countries explicitly criminalized people based on gender identity or expression. These laws existed at the national level in Guyana, Jordan, Lebanon, Malawi, Saudi Arabia, and the United Arab Emirates, and sub-nationally in Nigeria and Malaysia.19 Most laws explicitly targeted male-assigned individuals who dressed or “disguised” themselves as women, but some countries enacted more expansive provisions. For example, Guyana and Saudi Arabia criminalized “cross-dressing” regardless of birth-assigned sex, and some states in Malaysia adopted laws prohibiting female-assigned individuals from “posing” as men. Five additional countries implemented criminal provisions by 2021. The Gambia, Indonesia, Oman, and South Sudan incorporated laws targeting male-assigned individuals, while Kuwait’s law applied regardless of birth-assigned sex.20 p g Breaking down the TRIP score, ﬁgure 2 displays the proportion of countries for which each binary indicator equals one (as speciﬁed in table 2) in 2000 versus 2021. As this ﬁgure illustrates, advancements in transgender rights between these years are primarily a result of improvements in indicators pertaining to legal gender recognition and some anti-discrimination protections. More speciﬁcally, the proportion of countries with national laws allowing gender marker changes and those protecting against employment discrimination represent the most g g While uncommon, these laws appear to be resilient once in place. Decriminalization only occurred in one country throughout the period covered by the data. In Figure 1 Mean global and regional TRIP scores, 2000–2021 Figure 1 Mean global and regional TRIP scores, 2000–2021 .1 .2 .3 .4 .5 .6 TRIP Score 2000 2005 2010 2015 2020 Year Global E. Europe & C. Asia L. Amer. & Caribbean M. East & N. Africa Sub-Saharan Africa Asia & Pacific W. Europe & N. Amer. Figure 1 Mean global and regional TRIP scores, 2000–2021 September 2024 \| Vol. 22/No. 3 805 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Article \| A Global Analysis of Transgender Rights identity documents (ﬁgure 3). Most of these countries were in Europe. In 1972, Sweden became the ﬁrst European country to adopt a formal and nationally appli- cable law allowing this change.22 Germany, Italy, and the Netherlands followed suit in the 1980s, while Switzerland and Denmark adopted laws in 1993 and 2000, respec- tively. Figure 2 TRIP score indicators in 2000 versus 2021 Figure 2 TRIP score indicators in 2000 versus 2021 No direct crim. No indirect crim. GMC possible No physiological req. No psychological req. No divorce req. Nonbinary marker ADP - General ADP - Constitution ADP - Employment ADP - Education ADP - Healthcare ADP - Housing 0 .2 .4 .6 .8 1 Proportion of Countries 2000 2021 Note: Markers indicate the proportion of countries for which each binary indicator equals one in the respective year. g y The legal situation improved in most regions over the next 20 years. By 2021, 60 countries allowed legal gender marker changes (ﬁgure 4). Western Europe remained the front-runner in this category, with 19 of the recognizing countries in this region. New Zealand was the only other Western country with national recognition. While some states and provinces in the United States, Australia, and Canada allow gender marker changes, no nationally applicable laws or policies guarantee this right outside of travel documents (e.g., passports). Beyond the West, ten countries in Latin America, eight in Asia, and two in Africa adopted laws allowing gender marker changes by 2021. Note: Markers indicate the proportion of countries for which each binary indicator equals one in the respective year. Initially, all countries with legal recognition had at least one prohibitive requirement. This changed follow- ing the passage of the 2009 law in Uruguay, which did not include any physiological, psychological, or divorce/ single-status prerequisites.23 The law did however require people to undergo a review process with an expert team for at least two years. In 2012, Argentina became the ﬁrst country with legal gender recognition based entirely on self-determination.24 Over the next nine years, 14 additional countries allowed gender marker changes without any prohibitive requirements captured in the TRIP data.25 2018, the Caribbean Court of Justice ruled that section 153(1)(vlxii) of Guyana’s Summary Jurisdiction (Oﬀenses) Act was unconstitutional.21 This ruling nulli- ﬁed the criminalizing provision, even though parliament did not oﬃcially amend the law until 2021. Despite this progress, transgender people in Guyana remain at risk, especially given their lack of legal recognition and other protections. Indirect criminalization is far more widespread. In 2000, transgender people were susceptible to this type of criminalization in at least 112 countries. While this num- ber decreased slightly throughout the period, indirect criminalization remained present in 104 countries as of 2021. 806 Perspectives on Politics Figure 2 TRIP score indicators in 2000 versus 2021 The greater prevalence of indirect rather than direct criminalization in the dataset is in part attributable to the vast array of laws that countries can use for this purpose. Direct criminalization requires a speciﬁc type of law—one that explicitly criminalizes non-cisnormative identity or expression. However, indirect criminalization can occur through the presence and arbitrary application of a variety of laws policing other behaviors. For example, numerous countries target trans people through vague laws prohibit- ing “debauchery,” “hooliganism,” or “unnatural behavior” (Botha 2021; Egerton-Peters et al. 2018). Third-gender recognition continues to be rare. As of 2021, eight countries allowed individuals to select a non- binary gender on identity documents beyond passports.26 Interestingly, all four countries in Asia recognized non- binary markers following Supreme Court decisions. In contrast, the four countries in Europe and South America enacted laws independently of the courts. Further, the Asian countries were the ﬁrst to make these gender marker options available. While Uruguay and Argentina were early adopters of simpliﬁed gender recognition processes, they did not recognize nonbinary/third genders until 2018 and 2021, respectively. Criminalization l Several Eastern European and Central Asian coun- tries added legal gender recognition laws in the ﬁrst 10 years following the collapse of the Soviet Union. Beyond these regions, countries including Iran, Israel, Namibia, New Zealand, and Turkey had laws in place by 2000. No countries in the Americas had de jure gender recognition that year. Figure 2 TRIP score indicators in 2000 versus 2021 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press September 2024 \| Vol. 22/No. 3 807 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Comparing Transgender and LGB Rights in 2020 This section compares the rights countries oﬀered to sexual orientation and transgender minorities in 2020. To do so, I calculate LGB scores for each country using 11 indicators provided by ILGA World’s State-Sponsored Homophobia Report (Mendos et al. 2020). Following the TRIP method- ology, indicators equal one when a country protects a right on the national level.27 For example, the constitutional anti- discrimination indicator equals one if a country’s constitu- tion explicitly protects individuals based on sexual orienta- tion. I then added these values to create a composite score ranging from 0 to 11 (table 3). Comparing the two index scores within single countries also reveals important diﬀerences in the extent of legal rights available. In many cases, individual countries appeared to score similarly across the indexes in 2020 (ﬁgure 6). Countries less protective of one group tended to be less protective of the other. However, a few countries deviate considerably from this trend. As ﬁgure 5 high- lights, some cases like Pakistan, India, Austria, and South Africa perform disproportionately better on one of the indexes. These deviations emphasize the issues that may arise when measuring LGBT rights without incorpo- rating measures relevant to each group in the acronym. Before comparing the data, it is essential to note that the indicators in the TRIP and LGB indexes do not correspond with one another exactly. For example, the TRIP indicator capturing legal gender recognition is not directly compara- ble to any indicator in the LGB index. While this reinforces the point that separate data for LGB and trans rights is necessary, these diﬀerences make one-to-one comparisons diﬃcult. I convert countries’ raw scores on both indexes to a percentage to help account for any diﬀerences. Pakistan had the largest diﬀerence in scores on the two indexes. Following independence, the country retained the colonial provision criminalizing same-sex sexual activity (Mendos et al. 2020). Section 377 of the Penal Code continued to be active as of 2020, leaving sexual orientation minorities in danger of arrest (Human Dignity Trust 2022). This criminalization, paired with no legal recognition or protections, resulted in Pakistan scoring 0% on the LGB index during that year. At the start of the TRIP data, Pakistan performed poorly on indicators of trans rights as well. While there was no direct criminalization, the country did not oﬀer any trans-speciﬁc rights. TRIP versus LGB Scores In 2020, the average global score was 24.1% on the LGB index and 23.3% on the TRIP index. Though global performances nearly mirror one another, regional averages diﬀered more between the indexes (ﬁgure 5). These diﬀerences were most pronounced in regions with greater sexual orientation rights. For example, the average LGB score was over 13% higher than TRIP in Western Europe and North America. However, most regions scored higher on the TRIP index. Of these regions, Asia and the Paciﬁc exhibited the greatest diﬀerence, as TRIP was 9.1% higher than the LGB score. Sub-Saharan Africa scored the most similarly on both indexes, with a less than 1% diﬀerence. Anti-Discrimination Protections In 2000, 18 countries had national laws allowing individ- uals to change their gender markers on legal At the turn of the century, no countries oﬀered national anti-discrimination laws explicitly protecting gender https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Figure 3 National laws allowing gender marker change, 2000 Figure 4 National laws allowing legal gender marker change, 2021 Figure 3 National laws allowing gender marker change, 2000 Figure 3 Figure 3 National laws allowing gender marker change, 2000 Figure 3 National laws allowing gender marker change, 2000 gu e 3 National laws allowing gender marker change, 2000 Figure 4 National laws allowing legal gender marker change, 2021 Figure 4 National laws allowing legal gender marker change, 2021 g National laws allowing legal gender marker change, 2021 g National laws allowing legal gender marker change, 2021 identity, expression, or “reassignment.” The situation slowly improved over the period, with 39 diﬀerent coun- tries having at least one type of inclusive anti- discrimination law by 2021. In terms of the individual sectors, employment protections were the most common type. Thirty-ﬁve countries had these laws in place by the end of the period. Protections concerning the other spe- ciﬁc sectors were not as common but do not trail far behind. In 2021, 30 countries prohibited discrimination in education and 29 did so in healthcare and housing. All countries with one of these types of provisions during that year also provided employment protections. Constitutional protections are the least common type across the globe. In 2008, Ecuador became the ﬁrst country to incorporate “gender identity” as a protected class within a constitution. Bolivia, Cuba, Fiji, and Malta September 2024 \| Vol. 22/No. 3 807 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Article \| A Global Analysis of Transgender Rights have since followed suit, and remain as the only other countries to include explicit constitutional protections as of 2021. Interestingly, in every case except for Malta, the incorporation of gender identity as a protected class occurred with the adoption of entirely new constitutions. Malta is the only country that amended an existing constitution to add these types of protections. Comparing Transgender and LGB Rights in 2020 Further, trans indi- viduals were subject to arbitrary arrest through laws policing sexual activity and public obscenity (Botha 2021).28 The country only scored a 7.69% on the TRIP index in 2000. Table 3 LGB indicators for score calculations Indicators Score No direct criminalization (consensual same- sex sexual activity) 1 Same-sex marriage legal 1 Civil unions legal 1 Joint adoption for same-sex families legal 1 Second parent adoption for same-sex families legal 1 Constitutional anti-discrimination protections (sexual orientation) 1 Broad anti-discrimination protections (sexual orientation) 1 Employment anti-discrimination protections (sexual orientation) 1 Prohibition of hate crimes (sexual orientation) 1 Prohibition of incitement to hatred (sexual orientation) 1 Ban on conversion therapies (sexual orientation) 1 Total 11 Table 3 LGB indicators for score calculations Indicators Score No direct criminalization (consensual same- sex sexual activity) 1 Same-sex marriage legal 1 Civil unions legal 1 Joint adoption for same-sex families legal 1 Second parent adoption for same-sex families legal 1 Constitutional anti-discrimination protections (sexual orientation) 1 Broad anti-discrimination protections (sexual orientation) 1 Employment anti-discrimination protections (sexual orientation) 1 Prohibition of hate crimes (sexual orientation) 1 Prohibition of incitement to hatred (sexual orientation) 1 Ban on conversion therapies (sexual orientation) 1 Total 11 Table 3 LGB indicators for score calculations Indicators S The legal situation began to shift following an incident in the city of Rawalpindi. In January 2009, Rawalpindi police raided a wedding ceremony and arrested several gender diverse individuals (Pamment 2019; Redding 2018). The arrests sparked protests and ultimately culmi- nated in activists ﬁling a legal petition with the Supreme Court.29 Favorable decisions by the court, later coupled with the passage of laws such as the Transgender Persons (Protection of Rights) Act of 2018, granted the transgen- der community several legal rights. By 2020, Pakistan scored 84.62% on the TRIP index due to oﬀering com- prehensive anti-discrimination protections, nonbinary markers, and gender marker changes without prohibitive requirements. 808 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press The evolution of LGBT rights followed a similar path in India, resulting in considerably higher scores on the TRIP index. Like in Pakistan, a combination of Supreme Court cases and subsequent laws advanced the legal rights of transgender people. 30 As of 2020, India oﬀered some anti- discrimination protections and allowed gender marker changes once individuals satisﬁed the p ments. The country scored 61.54% on that year. Sexual orientation minoritie legal protections. Comparing Transgender and LGB Rights in 2020 While India decrimi same-sex sexual activity in 2018, the c protections for or, recognition of, Figure 5 TRIP and LGB index averages by region, 2020 0 .2 .4 .6 .8 Asia and Pacific W. Europe and N. America Sub-Saharan Africa Middle East and N. Africa L. American and the Caribbean E. Europe and C. Asia TRIP Score LGB Score Figure 6 TRIP versus LGB index scores, 2020 0 .2 .4 .6 .8 1 TRIP Score 0 .2 .4 .6 .8 1 LGB Score Note: Regression line with 90% conﬁdence interval. P-value < 0.001. R-squared = 0.5952. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press September 2024 \| Vol. 22/No. 3 809 Figure 5 TRIP and LGB index averages by region, 2020 Figure 5 TRIP and LGB index averages by region, 2020 0 .2 .4 .6 .8 Asia and Pacific W. Europe and N. America Sub-Saharan Africa Middle East and N. Africa L. American and the Caribbean E. Europe and C. Asia TRIP Score LGB Score Asia and Pacific Figure 6 TRIP versus LGB index scores, 2020 Figure 6 TRIP versus LGB index scores, 2020 TRIP versus LGB index scores, 2020 TRIP versus LGB index scores, 2020 0 .2 .4 .6 .8 1 TRIP Score 0 .2 .4 .6 .8 1 LGB Score Note: Regression line with 90% conﬁdence interval. P-value < 0.001. R-squared = 0.5952. 0 .2 .4 .6 .8 1 TRIP Score 0 .2 .4 .6 .8 1 LGB Score Note: Regression line with 90% conﬁdence interval. P-value < 0.001. R-squared = 0.5952. Note: Regression line with 90% conﬁdence interval. P-value < 0.001. R-squared = 0.5952. changes once individuals satisﬁed the prohibitive require- ments. The country scored 61.54% on the TRIP index in that year. Sexual orientation minorities enjoyed far fewer legal protections. While India decriminalized consensual same-sex sexual activity in 2018, the country still had no protections for or, recognition of, these individuals The evolution of LGBT rights followed a similar path in India, resulting in considerably higher scores on the TRIP index. Like in Pakistan, a combination of Supreme Court cases and subsequent laws advanced the legal rights of transgender people. 30 As of 2020, India oﬀered some anti- discrimination protections and allowed gender marker September 2024 \| Vol. 22/No. 3 809 https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Article \| A Global Analysis of Transgender Rights (Mendos et al. 2020). Overall, India scored 9.09% on the LGB index, 52.45% less than the country’s TRIP score. in the TRIP dataset, constitutions in Kosovo, Portugal, South Africa, and Sweden prohibit discrimination against sexual orientation but not gender identity. On the other end of the spectrum, Austria and South Africa are two cases where sexual orientation minor- ities had far greater legal rights than transgender individ- uals. Both countries scored 81.8% on the LGB index but only 23.1% on the TRIP index in 2020. Austria scored well on all LGB indicators except constitutional protec- tions and conversion therapy bans. Conversely, the coun- try performed poorly on all but three of the TRIP indicators. Austria did not directly or indirectly criminalize transgender people, but no anti-discrimination laws pro- tected these minorities. Figure 5 TRIP and LGB index averages by region, 2020 Further, individuals could change their legal gender markers only after meeting all prohibi- tive requirements.31 Austria also did not allow non binary gender markers. The measure of direct criminalization also demonstrates the wide gap in laws regarding these two groups.34 As of 2020, laws explicitly criminalizing consensual same-sex sexual activity existed in 59 of the 173 countries. On the other hand, only 12 countries speciﬁcally outlawed diverse gender identities or expressions. Though transgender peo- ple are vulnerable to indirect criminalization in countries prohibiting same-sex sexual activity, there are considerable diﬀerences in the formal laws explicitly policing groups within the LGBT community. However, the increase in direct criminalization captured by the TRIP data could indicate that this trend is slowly shifting. Hostile states may become more likely to target trans individuals directly as political debates surrounding their rights gain traction and visibility. Though South Africa did not ban conversion therapy or explicitly protect sexual orientation minorities against hate crimes, the country is one of the few to oﬀer constitutional protections to LGB individuals (Mendos et al. 2020). Altogether, South Africa scored positively on nine LGB indicators but provided few rights for trans people. No laws directly criminalized diverse gender identities or expres- sions, but trans individuals faced indirect criminalization through laws policing oﬀenses like public indecency and sex work.32 Further, South Africa allowed binary gender marker changes but conditioned these changes on physi- ological and psychological requirements.33 Finally, there were no national anti-discrimination protections for trans people. Potential Correlates of Transgender Rights This section uses the TRIP data to provide a preliminary look into the factors that may help explain variation in transgender rights throughout the world. Importantly, the analyses in this section are intended only as a ﬁrst step, rather than the ﬁnal word on the matter. Future research should build from these initial ﬁndings to further theorize about and empirically investigate why some countries oﬀer more rights than others. Since previous data limitations constrained trans-speciﬁc research, I draw from existing studies related to sexual orientation minorities to identify three potential correlates of transgender rights. These include regime type, economic development, and religion. Taken together, the four cases demonstrate the issues of using rights measures for one group as a proxy for the other. Countries do not inherently treat sexual orientation and transgender rights equally and conﬂating the two measures may misrepresent the actual domestic legal situation. Though the extent varies, score discrepancies are not unique to the four cases. Nearly 36% of all countries had a 10% or greater diﬀerence in their LGB and TRIP scores for 2020. More speciﬁcally, TRIP scores were higher by 10% in 24 countries, while LGB scores were higher in 39 countries during that year. g yp p g In terms of regime type, research suggests a positive relationship between democracy and sexual orientation rights (Dicklitch-Nelson et al. 2019; Encarnacion 2014; Sommer and Asal 2014). While democratic elections do not guarantee equal rights for all, certain features of liberal democracy help create conditions that are favorable to advancing these rights. For example, the greater respect for freedoms of assembly and expression in more democratic countries allows minorities, like LGBT people, to live more openly and collectively advocate for their rights to an extent unparalleled in nondemocracies (Encarnacion 2014). However, these freedoms may also empower polit- ical opponents who seek to restrict these rights, thus limiting the beneﬁts democracy provides. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press 810 Perspectives on Politics TRIP versus LGB Indicators This variable focuses more narrowly on the insti- tutional aspects of democracy and includes measures related to the competitiveness and openness of executive recruitment, constraints on the executive, and competi- tiveness of political participation. For easier comparisons with other estimates, I rescaled the variable so that values range between 0 and 1. As an additional robustness check, table A3 in the online appendix presents models with regime type measures from Regimes of the World (Luhrmann, Tannenberg, and Lindberg 2018) and Boix, Miller, and Rosato (2013, 2018).36 As a control, I include regional dummy variables, with Western countries serving as the base for comparison.38 This helps account for diﬀerences between countries that may stem from the inﬂuence of regional conditions or norms (Asal, Sommer, and Harwood 2012; Dicklitch- Nelson et al. 2019; Frank, Camp, and Boutcher 2010). Regional classiﬁcations are based on the politico- geographic regions provided by V-Dem (Coppedge et al. 2022). The TRIP scores (as percentages) serve as the dependent variable in all the models. Table 4 presents descriptive statistics for the key variables. Greater economic development has also been linked to more progressive sexual orientation rights (Badgett et al. 2014; Sommer and Asal 2014). Theoretically, as countries become more economically developed, they can turn their attention to “post-materialist” concerns such as expanding minority rights (Corrales 2017; Inglehart and Baker 2000). Higher levels of economic development also encourage more social tolerance towards minority groups, potentially providing more favorable conditions for rights advancements (Adamczyck and Pitt 2009). I use GDP per capita data from the World Bank (2022) to measure the level of economic development for each country-year. Following previous research, the models include the logged values of this measure given the skewed distribu- tion (Asal, Sommer, and Harwood 2012; Frank, Camp, and Boutcher 2010; Velasco 2018, 2020). As standard deviations in table 4 demonstrate, within- country variance is low for most variables due to their slow- moving nature. This has implications for which estimation strategy is most appropriate to use. For example, ﬁxed-eﬀects and random-eﬀects may produce unreliable estimates since these models take into account within-country variance (Beck 2001; Hill et al. 2020; Plümper and Troeger 2007). Therefore, I instead run one OLS model with country- clustered standard errors and another with panel-corrected standard errors (PCSE) to analyze the variation in transgen- der rights protections. TRIP versus LGB Indicators While the indexes do not overlap completely, they share a few comparable indicators that demonstrate the diver- gences in rights for the two groups. For example, nearly twice as many countries have anti-discrimination protec- tions for sexual orientation minorities. In 2020, 73 coun- tries had at least one type of anti-discrimination law covering sexual orientation, compared to just 39 countries with a similar law for gender identity minorities. Further, all countries with constitutional protections for transgen- der people also include provisions for sexual orientation. However, the same is not true in reverse. Of the countries I use two separate democracy measures to capture regime type, given the extensive and continued scholarly debate about how the concept should be measured (e.g., Alvarez et al. 1996; Boix, Miller, and Rosato 2013; Lindberg et al. 2014; Munck and Verkuilen 2002; Paxton 2000). For the ﬁrst measure, I use the electoral democracy index (EDI) from V-Dem (Coppedge et al. 2022). This index includes indicators concerning the freeness and fairness of elections, 810 Perspectives on Politics only provides estimates for every 10 years, I used the average change between decades to impute annual values. For a robustness check, tables A4 and A5 in the online appendix include models using data from the Religious Characteristics of States (RCS) Dataset (Brown and James 2017). Unlike Pew, the RCS provides annual-country year data without requiring additional calculations. However, I do not use this data for the main models because the dataset ends in 2015, which drops observations from the last six years of the TRIP data. The models utilizing RCS data include one variable measuring the overall percentage of the population that is religious (like Pew), and addi- tional variables disaggregating the percentage of adherents by Christianity, Islam, and non-religious in case diﬀer- ences persist between speciﬁc religions.37 suﬀrage, freedom of association, and freedom of expression. The EDI works well for this analysis because it provides a ﬁne-grained measure of democracy without including mea- sures related to transgender rights.35 Values for this variable range from 0 to 1, with higher values indicating greater levels of democracy. For the second measure, I use the revised combined Polity score (Polity2) from Polity V (Marshall and Jaggers 2020). https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press September 2024 \| Vol. 22/No. 3 811 TRIP versus LGB Indicators Mean Overall Between Within TRIP score 0 0.923 0.159 0.157 0.122 0.099 Electoral democracy 0.015 0.924 0.521 0.263 0.254 0.070 Polity2 (rescaled) 0 1 0.684 0.317 0.303 0.093 GDP per capita (logged) 4.603 11.803 8.275 1.558 1.489 0.479 Religiosity (%) 0.215 1 0.912 0.138 0.138 0.006 Table 5 Regressions of TRIP scores (1) (2) (3) (4) Models OLS OLS + PCSE OLS OLS + PCSE Electoral democracy 0.00815* 0.00815 (0.00244) (0.00334) Polity2 (rescaled) 0.00600* 0.00600* (0.00191) (0.00172) GDP per capita (log) 0.00120** 0.00120* 0.00187* 0.00187 (0.000504) (0.000701) (0.000607) (0.000770) Religiosity (%) −0.00318 −0.00318 −0.00588 −0.00588 (0.00675) (0.00953) (0.00736) (0.0105) Lagged TRIP score 0.989* 0.989* 0.989* 0.989* (0.00575) (0.0197) (0.00713) (0.0225) Regions Asia & Paciﬁc −0.01000* −0.01000 −0.00938 −0.00938 (0.00376) (0.00437) (0.00411) (0.00465) E. Europe & C. Asia −0.00511 −0.00511 −0.00442 −0.00442 (0.00359) (0.00448) (0.00393) (0.00480) L. America & the Caribbean −0.00852** −0.00852* −0.00678 −0.00678 (0.00379) (0.00462) (0.00414) (0.00493) Middle East & N. Africa −0.0146* −0.0146* −0.0142* −0.0142* (0.00339) (0.00488) (0.00355) (0.00527) Sub-Saharan Africa −0.0127* −0.0127 −0.0114* −0.0114 (0.00332) (0.00515) (0.00361) (0.00556) W. Europe & N. America Base Base Base Base Constant 0.00540 0.00540 0.00180 0.00180 (0.00857) (0.0139) (0.00956) (0.0152) Observations 3548 3548 3033 3033 Countries 172 172 164 164 R-Squared 0.927 0.927 0.907 0.907 Note: Models 1 and 3 are estimated using pooled OLS with country-clustered standard errors shown in parentheses. Models 2 and 4 are estimated using OLS with panel corrected standard errors shown in parentheses. * p < 0.1, Table 4 Descriptive statistics Standard Deviation Variable Min. Max. Mean Overall Between Within TRIP score 0 0.923 0.159 0.157 0.122 0.099 Electoral democracy 0.015 0.924 0.521 0.263 0.254 0.070 Polity2 (rescaled) 0 1 0.684 0.317 0.303 0.093 GDP per capita (logged) 4.603 11.803 8.275 1.558 1.489 0.479 Religiosity (%) 0.215 1 0.912 0.138 0.138 0.006 Table 5 Regressions of TRIP scores (1) (2) (3) (4) Models OLS OLS + PCSE OLS OLS + PCSE Electoral democracy 0.00815* 0.00815 (0.00244) (0.00334) Polity2 (rescaled) 0.00600* 0.00600* (0.00191) (0.00172) GDP per capita (log) 0.00120** 0.00120* 0.00187* 0.00187 (0.000504) (0.000701) (0.000607) (0.000770) Religiosity (%) −0.00318 −0.00318 −0.00588 −0.00588 (0.00675) (0.00953) (0.00736) (0.0105) Lagged TRIP score 0.989* 0.989* 0.989* 0.989* (0.00575) (0.0197) (0.00713) (0.0225) Regions Asia & Paciﬁc −0.01000* −0.01000 −0.00938 −0.00938 (0.00376) (0.00437) (0.00411) (0.00465) E. Europe & C. Asia −0.00511 −0.00511 −0.00442 −0.00442 (0.00359) (0.00448) (0.00393) (0.00480) L. TRIP versus LGB Indicators The PCSE models serve as a robust- ness check against potential heteroskedasticity that may bias results when using cross-national time series data (Beck and Katz 1995, 1996).39 All models include one-year lagged independent variables and a lagged dependent variable on the right-hand side to account for a country’s TRIP score in the previous year. Unlike democracy and economic development, existing studies suggest that religion impedes the advancement of sexual orientation rights (Corrales 2017; Dicklitch-Nelson et al. 2019). This negative relationship is perhaps unsur- prising since current interpretations of foundational texts across several major religions condemn same-sex sexual activity (Asal and Sommer 2017; Plante 2022). For exam- ple, in Christianity, passages in Leviticus (18:22 and 20:13) deem sexual relations between men as an “abomination” punishable by death (Plante 2022; Sanders 2009). Therefore, as traditional religious ideals concerning sexuality and gender identity exert a stronger inﬂuence over society and government, countries may be less likely to protect the rights of an unfavorable minority (Asal, Sommer, and Harwood 2012; Corrales 2017; Kollman 2007). Table 5 presents results from the main models in the analysis. In all models, both measures of democracy (EDI and Polity2) and GDP per capita share a positive and statistically signiﬁcant relationship with the TRIP score. In other words, greater levels of democracy and economic development appear to be associated with an increase in a country’s transgender rights. However, coeﬃcients for these variables are incredibly small, suggesting that the magnitude of this relationship is not substantial despite being signiﬁcant. Figure 7 demonstrates how an increase from the minimum to the maximum values for democracy or GDP per capita (logged) correspond with a less than one To measure religion, I use estimates from the Pew Research Center (2015) to calculate the percentage of a country’s population that identiﬁes as religious as opposed to unaﬃliated (Dicklitch-Nelson et al. 2019). Since Pew September 2024 \| Vol. 22/No. 3 811 Article \| A Global Analysis of Transgender Rights Table 4 Descriptive statistics Standard Deviation Variable Min. Max. 812 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press TRIP versus LGB Indicators The 812 Perspectives on Politics https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Figure 7 Predicted margins with 95% conﬁdence intervals .14 .15 .16 .17 TRIP Score 0 .1 .2 .3 .4 .5 .6 .7 .8 .9 1 Electoral democracy Pooled OLS - EDI .14 .15 .16 .17 TRIP Score 0 .1 .2 .3 .4 .5 .6 .7 .8 .9 1 Electoral democracy Panel Corrected SE - EDI .14 .15 .16 .17 TRIP Score 0 1 2 3 4 5 6 7 8 9 10 11 12 GDP per capita (log) Pooled OLS - GDP per capita .14 .15 .16 .17 TRIP Score 0 1 2 3 4 5 6 7 8 9 10 11 12 GDP per capita (log) Panel Corrected SE - GDP per capita onﬁdence intervals 5 .6 .7 .8 .9 1 emocracy S - EDI .14 .15 .16 .17 TRIP Score 0 .1 .2 .3 .4 .5 .6 .7 .8 .9 1 Electoral democracy Panel Corrected SE - EDI 6 7 8 9 10 11 12 apita (log) P per capita .14 .15 .16 .17 TRIP Score 0 1 2 3 4 5 6 7 8 9 10 11 12 GDP per capita (log) Panel Corrected SE - GDP per capita re 7 icted margins with 95% conﬁdence intervals .14 .15 .16 .17 TRIP Score 0 .1 .2 .3 .4 .5 .6 .7 .8 .9 1 Electoral democracy Pooled OLS - EDI g Predicted margins with 95% conﬁdence intervals .14 .15 .16 .17 TRIP Score 0 .1 .2 .3 .4 .5 .6 .7 .8 .9 1 Electoral democracy Panel Corrected SE - EDI Pooled OLS - EDI Panel Corrected SE - EDI Panel Corrected SE - EDI Electoral democracy Electoral democracy .14 .15 .16 .17 TRIP Score 0 1 2 3 4 5 6 7 8 9 10 11 12 GDP per capita (log) Panel Corrected SE - GDP per capita .14 .15 .16 .17 TRIP Score 0 1 2 3 4 5 6 7 8 9 10 11 12 GDP per capita (log) Pooled OLS - GDP per capita Panel Corrected SE - GDP per capita Pooled OLS - GDP per capita GDP per capita (log) GDP per capita (log) In the robustness models shown in the online appendix, the overall percentage of the population that is religious remains insigniﬁcant even when using the RCS data (table A4). TRIP versus LGB Indicators America & the Caribbean −0.00852** −0.00852* −0.00678 −0.00678 (0.00379) (0.00462) (0.00414) (0.00493) Middle East & N. Africa −0.0146* −0.0146* −0.0142* −0.0142* (0.00339) (0.00488) (0.00355) (0.00527) Sub-Saharan Africa −0.0127* −0.0127 −0.0114* −0.0114 (0.00332) (0.00515) (0.00361) (0.00556) W. Europe & N. America Base Base Base Base Constant 0.00540 0.00540 0.00180 0.00180 (0.00857) (0.0139) (0.00956) (0.0152) Observations 3548 3548 3033 3033 Countries 172 172 164 164 R-Squared 0.927 0.927 0.907 0.907 Note: Models 1 and 3 are estimated using pooled OLS with country-clustered standard errors shown in parentheses. Models 2 and 4 are estimated using OLS with panel corrected standard errors shown in parentheses. * p < 0.1, p < 0.05, * p < 0.01 Table 5 Regressions of TRIP scores 3 are estimated using pooled OLS with country-clustered standard errors shown in parentheses. Models 2 and 4 are S with panel corrected standard errors shown in parentheses. percentage point increase in TRIP scores. These ﬁndings also persist across the robustness models in the online appendix (table A3). measure in this analysis does not account for the extent to which religious beliefs inﬂuence individuals and their policy preferences, nor does it consider the role that religion plays in government. Though beyond the scope of this article, future research should carefully consider these additional ways religion and trans rights may be connected and how this diﬀers from the impact religion has on sexual orientation rights. While in the expected direction, coeﬃcients for the percentage of the population that is religious are not statistically signiﬁcant in any model in table 5. However, this null result does not necessarily imply the absence of a relationship between religion and transgender rights. TRIP versus LGB Indicators This ﬁnding also persists when distinguishing between speciﬁc religions. As table A5 shows, the variables measuring the percentage of the population that is Chris- tian, Muslim, or not religious are all insigniﬁcant. On the other hand, the EDI and GDP per capita variables con- tinue to be signiﬁcant and positive regardless of the religious measures. example, democracy and economic development were signiﬁcant throughout this analysis, as the literature on sexual orientation rights suggests. However, the magni- tude of their eﬀects is modest, meaning increases in these variables only correlate with minor rights improvements. This ﬁnding helps make sense of the incredible advance- ments in countries that have historically had low levels of democracy (e.g., Pakistan) or economic development (e.g., Argentina) but also raises questions about what factors are more pertinent to understanding variation in trans rights protections. Future research should build from these preliminary ﬁndings to further examine the role of these and other institutional and cultural factors in promoting transgender rights. Turning to the region dummies, coeﬃcients for Asia and the Paciﬁc, the Middle East and North Africa, and Sub-Saharan Africa were negative and statistically signiﬁ- cant in every model in table 5. Thus, countries in those regions are less protective of trans rights than countries in the Western Europe and North America region. Results for Latin America and the Caribbean were less consistent, as the signiﬁcance disappears in Models 3 and 4 where Polity2 served as the democracy measure. Finally, coeﬃ- cients for Eastern Europe and Central Asia were not signiﬁcantly diﬀerent from the West in any of the models. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press Conclusion Transgender rights are an important, but often neglected, component of cross-national LGBT studies in political science. Therefore, I introduced a new dataset that is among the ﬁrst to provide a collection of trans-speciﬁc measures on a global scale. The Trans Rights Indicator Project (TRIP) enables researchers to conduct more in-depth analyses of transgender rights separately from the broader LGBT community. TRIP also complements Though additional research is needed to investigate these relationships more adequately, the overall ﬁndings highlight potentially interesting diﬀerences between cor- relates of sexual orientation and transgender rights. For September 2024 \| Vol. 22/No. 3 813 Article \| A Global Analysis of Transgender Rights existing data sources that center around sexual orientation rights, allowing researchers to study the diﬀerences between these two groups. Overall, this article sheds light on an important but otherwise understudied topic in political science. Future work can utilize the TRIP data to conduct much-needed research on some of the most vulnerable minorities in the world. Beyond research, TRIP provides essential informa- tion to advocacy organizations and policymakers. Trans rights advocates can use this data to identify where trans minorities may be the most at risk and allocate resources accordingly. Finally, policymakers can beneﬁt from under- standing the diﬀerences between gender identity and sexual orientation rights to ensure that LGBT-based pol- icies adequately include transgender individuals. After introducing the data, I outlined the global evolu- tion of transgender rights from 2000 to 2021. Though many countries appeared to be improving, the average global TRIP score remained low at only 23.5% by the ﬁnal year covered in the data. The next section compared the TRIP and LGB indexes, revealing that countries are not always uniform in the rights they provide to sexual orientation and transgender minorities. Cases like Pakistan illustrate how conﬂating measures of LGBT rights can lead to a substantial misunderstanding of the actual legal rights that countries oﬀer. I concluded with an initial look into the eﬀects of regime type, economic development, and religion on countries’ trans rights pro- tections. The results suggest that greater levels of democ- racy and economic development are marginally associated with more rights, while the percentage of religious adher- ents in a country has no signiﬁcant impact. Acknowledgments The author would like to thank Amanda B. Edgell, Chris- topher W. Hale, Elif Kalaycioglu, Waleed Hazbun, Victor Asal, and the anonymous reviewers for their valuable feedback and support. While this article presents several preliminary ﬁndings, additional research is necessary to understand the com- plex dynamics that shape transgender rights globally. This research is especially critical as pushback against these rights spreads. Future research can use the TRIP data and build from this article to further investigate the causes and consequences of this resistance. Moreover, scholars should carefully consider the potential interplay between previous LGB(T) rights advancements and the subsequent backlash against trans rights. For example, the number of anti-trans bills in the United States nearly tripled the year after the Supreme Court ruling legalized same-sex marriage and roughly doubled following the Bostock decision that expanded anti-discrimination rights to LGBT individuals (Trans Legislation Tracker 2023). Future work should examine these connections more closely to determine whether and how certain advancements might inadvertently trigger a negative response to transgender rights. Supplementary Material To view supplementary material for this article, please visit http://doi.org/10.1017/S1537592723002827. https://doi.org/10.1017/S1537592723002827 Published online by Cambridge University Press 814 Perspectives on Politics Notes to rights (e.g., public opinion surveys) or data sources limited to a single country. 23 Law No. 18.620 Right to Gender Identity and to Change Name and Sex on Identiﬁcation Documents. 9 The ﬁve “LGBT” advocacy rights measures are not necessarily trans-speciﬁc as the variables appear to be duplicated in the gay rights and trans rights barome- ters. g 24 Identidad De Genero Ley 26.743. 25 Additional countries include Belgium, Brazil, Colombia, Costa Rica, Denmark, Ecuador, France, Iceland, Ireland, Luxembourg, Malta, Norway, Pakistan, and Portugal. 10 See ILGA Europe’s “Rainbow Map” at rainbow- europe.org. 11 See the “The Global Status of Transgender Rights, 2000–2021” section for speciﬁc examples of direct criminalization. 26 Latin America: Argentina and Uruguay. Asia: Bangladesh, India, Nepal, and Pakistan. Europe: Ice- land and Malta. 12 EU data from the European Union Agency for Fun- damental Rights (FRA) is available at: https://fra. europa.eu/en/data-and-maps/2020/lgbti-survey-data- explorer. For information on the U.S. survey, see James et al. (2016). 27 Following TRIP, criminalization accounts for national and subnational laws. 28 Penal Code (Act XLV of 1860), Section 377 and Section 294. 13 Yogyakarta Principles 12, 15, 16, and 17 refer to non- discrimination for employment, housing, education, and healthcare, respectively (ICJ 2007). 29 Khaki v. Rawalpindi 43/2009 (Supreme Court case). 30 NALSA v. Union of India 2014 (Supreme Court case); Transgender Persons Act (2019); Rules for Transgender Persons Act (2020). 14 Full text of the law available from Rainbow Europe is available at https://rainbow-europe.org/#8650/0/0. 31 The Civil Status Act (as amended in 2016). 15 The full text of Bolivia’s (2009) constitution is avail- able from the Constitute Project, https://www. constituteproject.org/constitution/Bolivia_2009? lang=en. 32 Sexual Oﬀences Act 23 of 1957, Sections 12A and 19-20. 33 Alteration of Sex Description and Sex Status Act 49 of 2003. 16 Despite the medicalization the term imposes, I include gender “reassignment” given the possible usage of the word in laws. 34 Comparisons of indirect criminalization are not pos- sible since the LGB index does not include this indicator, and the TRIP indicator includes laws criminalizing same-sex sexual activity. 17 Organizations are listed in alphabetical order. 17 Organizations are listed in alphabetical order. g 35 For example, Freedom House (2022) includes con- siderations of LGBT groups in their political rights and civil liberties scores. Notes g p 18 “Transsexual” traditionally refers to someone “who sought medical interventions to change their physical bodies (that is, their ‘sex’),” whether that be through surgery, hormone replacement, or other means (Stryker 2008, 18). “Transvestite” and “cross-dresser” evolved to carry similar meanings, typically describing “people who wear gender-atypical clothing but do not engage in other kinds of bodily modiﬁcation” (Stryker 2008, 17). 36 Regimes of the World classiﬁes regimes as closed autocracies, electoral autocracies, electoral democra- cies, or liberal democracies. Boix, Miller, and Rosato (2013) provide a binary classiﬁcation distinguishing between democratic and non-democratic regimes. 37 Since LGB-rights literature primarily focuses on Christianity and Islam, I include these measures in the robustness models. Future research should extend these analyses to incorporate additional religions. 19 Guyana: Summary Jurisdiction (Oﬀenses) Act, Section 153(1)(xlvii); Jordan: Penal Code, Article 307; Lebanon: Penal Code, Article 521; Malawi: Penal Code, Section 180(g); Malaysia: state-enacted Islamic laws (varies by state); Nigeria: Sharia Penal Code (various states), Section 376(2e-f); Kano State Prostitution and Other Immoral Acts (Prohibition) Law, Section 9; Saudi Arabia: national implementa- tion of Sharia law; United Arab Emirates: Penal Code, Article 359. 38 “Western” countries are those V-Dem codes as West- ern Europe and North America, which includes Australia and New Zealand. 38 “Western” countries are those V-Dem codes as West- ern Europe and North America, which includes Australia and New Zealand. 39 Tests for serial correlation and multicollinearity determine that neither issue is present in the OLS model. Results from these tests are available in tables A1 and A2 in the online appendix. 20 Indonesia: see Chiam et al. (2020) and Egerton-Peters et al. (2018); Kuwait: Penal Code, Article 198; Oman: Penal Code, Article 266; South Sudan: Penal Code, Section 379; The Gambia: Criminal Code, Section 167. Notes 1 LGBT stands for lesbian, gay, bisexual, and trans- gender. 2 While an essential methodological task on its own, these “merely descriptive” analyses also lay the groundwork for future studies to investigate causal relationships concerning transgender rights (Gerring 2012). 3 People who are intersex are born with some combi- nation of biological sex characteristics that do not fall exclusively in the category of “male” or “female” (Stryker 2008). y 4 Scholars and medical practitioners alike have long debated the distinction between sex and gender iden- tity, but those arguments fall beyond the scope of this article. For further discussion of the conceptual dif- ferences and evolution of the terminology, see Beemyn (2014), Hines (2020), Meyerowitz (2002), and Stry- ker (2008). p g g One limitation of the TRIP dataset is that it currently only includes legal indicators. As discussed, measures capturing societal violence and discrimination would allow for a more complete understanding of transgender rights, but this data is limited and suﬀers from underreporting. Further, laws still provide crucial insight into the condi- tions individuals may face within a country (Htun and Weldon 2012; Velasco 2018). In the future, I plan to build on the TRIP dataset by using expert surveys to capture de facto measures. Additionally, I plan to expand on the current set of legal measures to better capture the wide range of rights relevant to trans individuals. For example, future versions of the dataset will ideally include indicators pertaining to parental rights (e.g., adoption, surrogacy), eligibility for military service, and trans- speciﬁc hate crime protections. 5 “Transgender” also includes non-binary or third- gender identities (i.e., gender identities that fall out- side of or between male and female). 5 “Transgender” also includes non-binary or third- gender identities (i.e., gender identities that fall out- side of or between male and female). 6 As with gender identity, language describing sexual orientation is diverse, and examples within this article may be representative of only one understanding of these concepts. p 7 Bostock v. Clayton County, Georgia, 590 U.S., 140 S. 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https://openalex.org/W2782376463	https://europepmc.org/articles/pmc5800933?pdf=render	English	null	Association between PIK3CA alteration and prognosis of gastric cancer patients: a meta-analysis	Oncotarget	2,018	cc-by	4,915	ABSTRACT Background: Increasing evidence suggests that dysregulation of phosphatidylinositol-4, 5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) plays an important role in carcinogenesis. However, the relationship between PIK3CA expression and gastric cancer (GC) prognosis remains controversial. Methods: We searchedPubMed, Embase, Web of Science, and the Cochrane Library databases for relevant studies up to June 30, 2017. Primary outcomes were hazard ratio (HR), odds ratio (OR), and 95% confidence intervals (CI) for association with overall survival and clinicopathological features. Results: Eleven studies comprising 2481 GC patients were analyzed. Pooled analysis showed that PIK3CA upregulation was significantly associated with worse overall survival (HR = 1.79, 95% CI 1.42–2.27, p< 0.001) at the protein (HR = 1.94, 95% CI 1.52–2.47, p< 0.001) but not the gene (HR = 1.57, 95% CI 0.92–2.69, p= 0.097) level. PIK3CA gene mutation did not correlate with overall survival (HR = 1.05, 95% CI 0.83–1.34, p= 0.666) but was significantly associated with poor tumor differentiation (OR = 0.37, 95% CI 0.17–0.76, p= 0.011). Conclusion: High PIK3CA protein expression predicted poor prognosis in GC, whereas PIK3CA gene amplification or mutation did not. Moreover, PIK3CA mutation was an indicator of poorly differentiated tumors. Association between PIK3CA alteration and prognosis of gastric cancer patients: a meta-analysis y Hua Li1,, Shubo Chen2,, Hui Li3,, Jianxin Cui4,, Yunhe Gao4, Dianchao Wu1, Shangfeng Luan1, Yan Qin1, Tongshan Zhai1, Dengxiang Liu5 and Zhibin Huo1 1Department of Surgical Oncology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 2Department of Surgical Urology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 3Department of General Surgery, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 4Department of General Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, China 5Institute of Cancer Control, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China These authors have contributed equally to this work Published: January 02, 2018 Published: January 02, 2018 Copyright: Li et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Copyright: Li et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. www.impactjournals.com/oncotarget/ www.impactjournals.com/oncotarget/ INTRODUCTION [2]. Therefore, understanding the biological alterations associated with GC, especially those leading to the dysregulation of signaling pathways, might help to predict patient prognosis and identify novel therapeutic targets. Gastric cancer (GC) is the fourth most common malignant neoplasm worldwide, and it was estimated to be the second leading cause of cancer-related death in 2014 [1]. Despite recent breakthroughs in the diagnosis and treatment of GC, its prognosis remains unfavorable The phosphatidylinositol 3-kinase (PI3K) pathway is one of the most commonly activated and altered signaling pathways in cancer, including GC [3–5]. Since Association between PIK3CA alteration and prognosis of gastric cancer patients: a meta-analysis y cancer patients: a meta analysis Hua Li1,, Shubo Chen2,, Hui Li3,, Jianxin Cui4,, Yunhe Gao4, Dianchao Wu1, Shangfeng Luan1, Yan Qin1, Tongshan Zhai1, Dengxiang Liu5 and Zhibin Huo1 1Department of Surgical Oncology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 2Department of Surgical Urology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 3Department of General Surgery, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 4Department of General Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, China 5Institute of Cancer Control, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China These authors have contributed equally to this work Correspondence to: Zhibin Huo, email: huozhibinxt@126.com Dengxiang Liu, email: rmyy666@163.com Keywords: PIK3CA; gastric cancer; prognosis; alteration; meta-analysis Received: August 09, 2017 Accepted: December 26, 2017 Published: January 02, 2018 Copyright: Li et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Hua Li1,, Shubo Chen2,, Hui Li3,, Jianxin Cui4,, Yunhe Gao4, Dianchao Wu1, Shangfeng Luan1, Yan Qin1, Tongshan Zhai1, Dengxiang Liu5 and Zhibin Huo1 1Department of Surgical Oncology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 2Department of Surgical Urology, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 3Department of General Surgery, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China 4Department of General Surgery, Chinese People’s Liberation Army General Hospital, Beijing 100853, China 5Institute of Cancer Control, Affiliated Xing Tai People Hospital of Hebei Medial University, Xingtai 054001, China *These authors have contributed equally to this work Correspondence to: Zhibin Huo, email: huozhibinxt@126.com Dengxiang Liu, email: rmyy666@163.com Keywords: PIK3CA; gastric cancer; prognosis; alteration; meta-analysis Received: August 09, 2017 Accepted: December 26, 2017 Published: January 02, 2018 Copyright: Li et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. RESULTS the PI3K pathway plays an essential role in several cellular processes, including cell growth, metabolism, and survival, it is not surprising that its dysregulation is also of critical importance to pathological processes such as the development, progression, and metastasis of cancer [6, 7]. A key step in the PI3K/AKT pathway is the generation of phosphatidylinositol-3, 4, 5-trisphosphate (PIP3) by PI3K. The PI3K family exists as three subfamilies, one of which is composed of a p110 catalytic subunit coupled to a regulatory subunit. p100α, the gene product of phosphatidylinositol-4, 5-bisphosphate3-kinase, catalytic subunit alpha (PIK3CA), is of particular importance in signaling through the canonical PI3K pathway [8, 9]. Recently, large-scale next-generation sequencing studies have identified mutations in PIK3CA in Epstein–Barr virus (EBV)-associated and microsatellite instability (MSI)- associated GCs [10]. However, the prognostic value of PIK3CA alterations at both the protein and transcriptional levels in GC patients remains unclear and controversial. www.impactjournals.com/oncotarget www.impactjournals.com/oncotarget Oncotarget 7651 Eligible studies and general characteristics A total of 582 studies were retrieved from a search of PubMed, Embase, Web of Science, and Cochrane Library online databases using established strategies (see Materials and Methods). After a review of the titles and abstracts, 167 duplicates and 54 non-English language studies were excluded. Of the remaining 361 studies, 235 were excluded because they did not meet the eligibility criteria of our analysis. Another 44 studies did not report patient survival, 51 focused on components of the PI3K/ AKT pathway other than PIK3CA, and 19 did not examine PIK3CA expression in cancer tissues. Finally, a total of 11 studies were included in our meta-analysis (Figure 1). The general characteristics of the included studies are presented in Table 1. The 11 studies comprised 2481 patients for pooling analysis (median 208, range 79–568). Two studies were from Europe (UK and Italy) and the remaining nine were from East Asia (four from China, two from Japan, two from Korea, and one from Singapore). The various techniques used for the qualitative and Therefore, we conducted a meta-analysis of published studies to elucidate the precise relationship between PIK3CA dysregulation and the prognosis and other clinicopathological features of GC patients. Figure 1: Flow chart of study selection process. Figure 1: Flow chart of study selection process. www.impactjournals.com/oncotarget Oncotarget 7652 Table 1: Major characteristics of the selected studies Table 1: Major characteristics of the selected studies No. Eligible studies and general characteristics FC: figure calculation; HR: hazard ratio; IHC: immunohistochemistry; M/F: male to female ratio; M: multivariate analysis; NOS: Newcastle–Ottawa Scale; NR: not reported; qPCR: quantitative polymerase chain reaction; U: univariate analysis; R0: radical resection; R1: palliative surgery. 1.94, 95% CI 1.52–2.47, p< 0.001) but not PIK3CA gene amplification (HR = 1.57, 95% CI 0.92–2.69, p= 0.097) (Figure 2). quantitative detection of PIK3CA included polymerase chain reaction (PCR) followed by direct sequencing or pyrosequencing of amplicons, quantitative real-time PCR (qPCR), and immunohistochemistry (IHC) to detect protein. All studies were scored above 5 stars except Y Sukawa’s and M Liang’s reports according the Newcastle– Ottawa Scale (NOS) [22], which indicated a relatively high risk of bias [23, 24] (Supplementary Table 1). ( g ) When it refers to the different treatment settings, subgroup analysis showed that in the radical resection and palliative surgery mixed group [12, 17, 19], the association between PIK3CA overexpression and overall survival remains significant (HR = 2.13, 95% CI 1.67–2.72, p < 0.001), while in the radical resection-only group [15, 16, 20], it was not significant (HR = 1.25, 95% CI 0.86-1.83, p=0.246), which might indicate a more predominant role of PIK3CA overexpression predication on survival in those patients with late-stage diseases. Association between PIK3CA overexpression in GC tissue and patient prognosis Six studies provided sufficient data to analyze the relationship between overall survival (OS) and PIK3CA expression level. The pooled analysis showed that high PIK3CA expression was associated with worse OS (hazard ratio [HR] =1.79, 95% confidence intervals [CI] 1.42– 2.27, p < 0.001). Although no significant heterogeneity was observed (inconsistency index [I2] = 17.9%, p= 0.297), judging the presence of potential bias sources, a random-effects model was applied to make the conclusion more conservative and reliable [25] (Figure 2). Furthermore, the associations remained significant in both the univariate (HR=1.74, 95%CI 1.31-2.33, p < 0.001) and multivariate (HR=1.89, 95%CI 1.01-3.53, p=0.046) analysis models (Table 2, Left section). Eligible studies and general characteristics First author Year Number Sex(M/F) Age Region TNM Stagea Alteration type Detection method Treatment Follow up HR estimate NOS score 1 S Barbi [11] 2010 264 175/89 67.4(median) Italy I-IV Gene mutation(exon 9,20) Direct sequencing R0 Only NR Provided(M) 7 2 J Shi [12] 2012 131 102/29 NR China I-IV Gene mutation(exon 9,20) and amplification Direct sequencing and qPCR R0/R1 NR Provided(M) 7 3 Y Sukawa [13] 2012 231 157/74 71(25-91) Japan IB-IV Gene mutation(exon 9,20) Pyrosequence NR NR FC 5 4 AFC Okines [14] 2013 337(303)b 263/74 63(median) UK I-IIIC Gene mutation(exon 9,20) Direct sequencing R0 Only/ R0+Chemotherapy NR FC 9 5 ML Chong [15] 2013 79 64/15 53% (≥69) Singapore I-IV Gene amplification qPCR R0 Only 15(1- 131) Provided(U) 8 6 H Lee [16] 2015 110 83/27 50% (≥60) Korea I-III Gene mutation (exon9,20) and Amplification Direct sequence and qPCR R0 Only 82.2(3.7- 158.8) FC 7 7 M Liang [17] 2015 107 NR NR China I-IV Protein overexpression IHC R0/R1 NR FC 5 8 K Harada [18] 2016 208 148/60 NR Japan I-IV Gene mutation(exon 9,20) Pyrosequencing R0 Only 60 FC 7 9 M Dong [19] 2016 568 396/172 52.3% (≥60) China I-IV Protein overexpression IHC R0/R1 37.2(0- 93.8) FC 7 10 SH Jang [20] 2016 178 127/51 68%(≥55) Korea I-IV Protein overexpression IHC R0+Chemotherapy 58(0-95) Provided(M) 7 11 JW Kim [21] 2017 302 202/100 20.2%(≥70) Korea I-IV Gene mutation qPCR(exon 1,4,7,9,20) R0/R1 NR FC 7 aAmerican Joint Committee on Cancer 7th edition TNM stage system. b The actual number of patients analyzing PIK3CA status and survival was 303. FC: figure calculation; HR: hazard ratio; IHC: immunohistochemistry; M/F: male to female ratio; M: multivariate analysis; NOS: Newcastle–Ottawa Scale; NR: not reported; qPCR: quantitative polymerase chain reaction; U: univariate analysis; R0: radical resection; R1: palliative surgery. aAmerican Joint Committee on Cancer 7th edition TNM stage system. b The actual number of patients analyzing PIK3CA status and survival was 303. FC: figure calculation; HR: hazard ratio; IHC: immunohistochemistry; M/F: male to female ratio; M: multivariate analysis; NOS: Newcastle–Ottawa Scale; NR: not reported; qPCR: quantitative polymerase chain reaction; U: univariate analysis; R0: radical resection; R1: palliative surgery. aAmerican Joint Committee on Cancer 7th edition TNM stage system. b The actual number of patients analyzing PIK3CA status and survival was 303. DISCUSSION The PI3K/AKT pathway is an important therapeutic target because its components, including PIK3CA, are frequently activated in cancer cells; indeed, several PI3K pathway inhibitors are currently in clinical trials [26, 27]. Therefore, identifying patients who might benefit from targeted therapy is of great importance [28, 29]. An increasing number of studies are focused on PIK3CA dysregulation and its potential impact in GC, one example being the MAGIC trial [30], highlighting the urgent need to elucidate more precisely the contribution of PIK3CA alterations to GC. To the best of our knowledge, this meta-analysis of 11 studies comprising 2481 GC patients represents the first comprehensive systematic evaluation of the prognostic value of PIK3CA alterations detected at the protein and gene levels. The results suggest that PIK3CA could represent a useful biomarker for predicting patient survival. Association between PIK3CA dysregulation and clinicopathological features of GC patients The associations between PIK3CA dysregulation (overexpression and mutation) and major clinicopathological features are shown in Table 3. Of the characteristics analyzed, the only significant association was between the presence of mutated PIK3CA and tumor differentiation grade (well/moderate vs. poor/ undifferentiated, odds ratio [OR] 0.37, 95% CI 0.17–0.79, p= 0.011). Association between PIK3CA mutation in GC tissue and patient prognosis A total of seven studies were pooled for the analysis of the effect of PIK3CA mutation on GC patient prognosis. The results showed that the presence of mutations (mainly in exons 9 and 20) did not correlate significantly with OS (HR = 1.05, 95% CI 0.83–1.34, p = 0.666) (Figure 3). We then performed a subgroup analysis of studies that examined overexpression at the gene and protein level. The results showed that OS was significantly associated with PIK3CA protein overexpression (HR = www.impactjournals.com/oncotarget Oncotarget 7653 mutation studies, p= 0.881 for Begg’s test and p = 0.22 for Egger’s test. A subgroup analysis was also conducted due to the potential existence of study bias (Table 2, Right section). However, no obvious heterogeneity was observed and the association remains insignificant either in the subgroup of different statistical analysis or HR evaluation methods, which indicated a stable meta-analysis result within these subgroups. Publication bias Potential publication bias was assessed in the association of OS with PIK3CA alteration type. As shown in Figure 4, no significant bias was detected in the included studies. For the overexpression studies, p= 0.260 for Begg’s test and p= 0.271 for Egger’s test, and for the In theory, alterations in PIK3CA activity or expression could lead to downstream activation of PI3K/ AKT pathway and promote the growth and invasion of GC cells [7, 31]. Our pooled analysis showed that Figure 2: Forest plot of hazard ratios for overall survival of gastric cancer patients and PIK3CA overexpression stratified by PIK3CA alteration measurement. Figure 2: Forest plot of hazard ratios for overall survival of gastric cancer patients and PIK3CA overexpression stratified by PIK3CA alteration measurement. www.impactjournals.com/oncotarget Oncotarget 7654 Table 2: Subgroup analysis of PIK3CA status and patients’ survival Subgroup PIK3CA overexpression PIK3CA mutation Studies (Patients) HR (95%CI) Heterogeneity Studies (Patients) HR (95%CI) Heterogeneity I2 (%) P I2 (%) P Method IHC 3(853) 1.94(1.52-2.47) 0 0.507 / / / / qPCR and direct sequence 3(320) 1.57(0.92-2.69) 49.8 0.137 5(1110) 1.04(0.79-1.38) 0.0 0.840 Pyrosequence / / / / 2(439) 1.10(0.68-1.78) 0.0 0.476 Treatment R0 3(367) 1.25(0.86-1.83) 0 0.850 4(885) 0.99(0.71-1.38) 0.0 0.889 R0/R1 3(806) 2.13(1.67-2.72) 0 0.796 2(433) 1.07(0.69-1.65) 0.0 0.351 NR / / / / 1(231) 1.27(0.68-2.37) / / Race Mongolian 6(1173) 1.79(1.42-2.27) 17.9 0.297 5(982) 1.08(0.79-1.47) 0 0.845 Caucasian / / / / 2(567) 1.02(0.69-1.51) 0 0.466 Analysis method Multivariate 2(309) 1.89(1.01-3.53) 49.4 0.160 2(395) 1.17(0.78-1.77) 0 0.453 Univariate 4(864) 1.74(1.31-2.33) 26.5 0.253 5(1154) 1.00(0.74-1.35) 0 0.906 HR evaluation Provided 3(388) 1.72(1.11-2.64) 23.1 0.272 2(395) 1.17(0.78-1.77) 0 0.453 Calculated 3(785) 1.80(1.28-2.53) 40.5 0.186 5(1154) 1.00(0.74-1.35) 0 0.906 HR: hazard ratio; R0: radical resection; R1: palliative surgery; NR: not reported; CI: confidence interval; I2: inconsistency index. Table 2: Subgroup analysis of PIK3CA status and patients’ survival Figure 3: Forest plot of hazard ratios for overall survival of gastric cancer patients with PIK3CA gene mutations. Figure 3: Forest plot of hazard ratios for overall survival of gastric cancer patients with PIK3CA gene mutations. www.impactjournals.com/oncotarget Oncotarget 7655 Table 3: Meta-analysis of the association between PIK3CA alterations and clinicopathological features of gastric cancer patients PIK3CA overexpression Stratification No. of studies No. Publication bias of patients Pooled OR (95%CI) Heterogeneity I2(%) P-value Gender (M/F) 3 877 1.03 (0.77-1.39) 0 0.380 Lymph node metastasis (N0/N1-3) 3 877 2.23 (0.53-9.36) 91.1 <0.001 Distant metastasis (M1/M0) 2 1326 3.11 (0.35-27.42) 79 0.029 TNM stage (I+II/III+IV)a 3 877 0.52 (0.09-2.96) 95 <0.001 Differentiation (Well+Moderate/Poor) 2 308 1.31 (0.20-8.74) 88 0.004 Tumor size (<5cm vs ≥5cm) 2 699 4.43 (0.55-35.43) 85.8 0.008 Lauren classification (Intestinal/diffuse) 2 661 0.97 (0.16-6.04) 95.8 <0.001 PIK3CA mutation Stratification No. of studies No. of patients Pooled OR (95%CI) Heterogeneity I2(%) P-value Gender (M/F) 6 1200 0.93 (0.64-1.36) 0 0.822 Lymph node metastasis (N0/N1-3) 6 1183 0.95 (0.63-1.42) 0 0.762 TNM stage (I+II/III+IV)a 3 614 0.66 (0.39-1.12) 0 0.511 Differentiation (Well+Moderate/Poor) 2 386 0.37 (0.17-0.79) 0 0.522 Lauren classification (Intestinal/diffuse) 5 1009 0.76 (0.41-1.43) 52.7 0.076 Microsatellite instability (MSI/MSS) 2 566 0.68 (0.15-3.06) 82.9 0.016 aAmerican Joint Committee on Cancer 7th edition TNM stage system. CI: confidence interval; I2: inconsistency index; MSI: microsatellite instability. MSS: microsatellite stability; OR: odds ratio. PIK3CA dysregulation is associated with poorer tumor differentiation and worse OS among GC patients, indicating that PIK3CA is a prognostic biomarker for GC patients. However, the significant association between PIK3CA mutation and tumor differentiation should be interpreted with caution because of the small sample size (two studies comprising 386 patients) and need more studies to confirm. Furthermore, our results showed no significant association between PIK3CA mutational status and patient prognosis. This might be attributed to the fact that gene mutation of a single component does not necessarily reflect the status of an entire signaling pathway in terms of dysregulation and functional impact. Therefore, our results suggest that the detection of gene Figure 4: Funnel plot evaluation of potential publication bias on the effect of PIK3CA overexpression (A) and mutation (B) on overall survival of gastric cancer patients. Figure 4: Funnel plot evaluation of potential publication bias on the effect of PIK3CA overexpression (A) and mutation (B) on overall survival of gastric cancer patients. www.impactjournals.com/oncotarget www.impactjournals.com/oncotarget Oncotarget 7656 amplification and/or protein overexpression is a more direct and promising strategy to predict GC patient prognosis than gene mutation analysis. neoplasm” and “PIK3CA” or “phosphatidylinositol-4, 5-bisphosphate 3-kinase, catalytic subunit alpha” and “prognosis” or “survival.” Article language was limited to English. The reference section of all relevant articles was examined to identify additional related studies. Publication bias Two researchers (HL and YG) independently assessed the eligibility of the identified studies, and disagreements were resolved by discussion or consultation with a third researcher. The most recent NCCN guidelines for gastric cancer have described MSI and EBV status as potential molecular biomarkers for targeting treatment strategies in GC patients [32]. Although mutant PIK3CA is more common in MSI than in other GC types [10], our analysis failed to confirm this, possibly because of the small number of studies that analyzed MSI association (two studies comprising 566 patients). There were also insufficient data to analyze the association between PIK3CA mutation and EBV positivity, so further studies are necessary to evaluate this relationship. Data extraction The following items were extracted from the included studies: study title, name of the first author, publication year, region, PIK3CA alteration type, detection method, and clinicopathological characteristics (i.e., gender, TNM stage, and tumor differentiation). The HRs, ORs, and 95% CIs for the association of PIK3CA status with OS and clinicopathological features were collected from the studies. If HRs or ORs were not directly reported, survival data were calculated using the methods described by Parmar et al. [33] and Tierney et al. [34]. In conclusion, we found that overexpression of PIK3CA protein, but not the presence of PIK3CA gene amplification or mutations, predicted worse prognosis for GC patients. However, our findings, as well as other important variables such as the association of PIK3CA alterations with GC molecular subtypes, require further verification in more rigorous studies with consistent and standardized methodology. Inclusion criteria Studies were selected according to the following inclusion criteria: (1) inclusion of pathologically confirmed GC patients; (2) focus on PIK3CA alterations, at either the protein or gene level; and (3) inclusion of survival data according to the intra-tumor PIK3CA alteration status. Several limitations to our study should be noted. First, although we observed no obvious heterogeneity (Figures 2 and 3), the studies used different designs, PCR primers, and IHC antibodies, which might have contributed to heterogeneity. Therefore, a random-effects model was applied to make our results more conservative. Second, we obtained insufficient data to analyze the relationship between PIK3CA and disease-free survival, which reduces the predictive value of PIK3CA status on the outcomes of GC patients. Third, the included studies did not investigate the relationship between PIK3CA gene amplification and protein overexpression, which might be critical for selecting the appropriate method of analysis for different GC patients. Furthermore, association analysis of some clinicopathological features (Table 3) included only two to three studies, and therefore should be interpreted with caution; additional studies are required to confirm the conclusion. It is also noticeable that the patients from the MAGIC trial of AFC Okines’ study included a small portion of esophageal cancer patients (38 out of 337), which might add to the bias of our results. Finally, we limited the analysis to English language publications, which might have led to selection bias in our meta- analysis. Quality assessment The quality of the included studies was assessed independently by two researchers using the Newcastle– Ottawa Scale [22]. This scale is an eight-item instrument that assesses patient population and selection, study comparability, and outcomes. We considered a study high- quality if it was awarded six or more stars [23]. Exclusion criteria The exclusion criteria were (1) studies published in languages other than English, (2) replicate or overlapping publications, (3) analysis of only cancer cell lines, and (4) studies with a small sample size (≤20 patients). Literature search Meta-analysis was performed using Stata version 12.0 software (StataCorp, College Station, TX, USA). A significant heterogeneity was observed when P < 0.05 or I2 > 50%, and a random-effects model was then applied. Otherwise, a fixed-effects model was used. The search was conducted by consulting PubMed, Embase, Web of Science, and Cochrane Library online databases from inception to June 30, 2017. The search terms were “gastric/stomach cancer” or “gastric/stomach www.impactjournals.com/oncotarget FUNDING 14. Okines AF, de Castro DG, Cunningham D, Chau I, Langley RE, Thompson LC, Stenning SP, Saffery C, Barbachano Y, Coxon F, Middleton G, Ferry D, Crosby T, et al. Biomarker analysis in oesophagogastric cancer: results from the REAL3 and TransMAGIC trials. Eur J Cancer. 2013; 49:2116-25. This work was supported by Municipal Science funds of Xingtai (No. 2015ZC202). ETHICAL APPROVAL AND INFORMED CONSENT 13. Sukawa Y, Yamamoto H, Nosho K, Kunimoto H, Suzuki H, Adachi Y, Nakazawa M, Nobuoka T, Kawayama M, Mikami M, Matsuno T, Hasegawa T, Hirata K. Alterations in the human epidermal growth factor receptor 2-phosphatidylinositol 3-kinase-v-Akt pathway in gastric cancer. World J Gastroenterol. 2012; 18:6577-86. No studies with human participants were performed by any of the authors of this study. All the analyses were based on previously published studies. www.impactjournals.com/oncotarget Oncotarget 7657 Author contributions 7. Matsuoka T, Yashiro M. The Role of PI3K/Akt/mTOR Signaling in Gastric Carcinoma. Cancers (Basel). 2014; 6:1441-63. Z.H and D.L. design this study. J.C and Y.G. searched, evaluated and collected the studies. D.W. and S.L extracted the data. H.L and S.C performed the meta- analysis. Hui L. and Y.Q. helped the whole process. Y.Q and T.Z. contributed to the study enrollment and revision. H.L and Y.G wrote the manuscript. All authors reviewed this manuscript. 8. Cantley LC. The phosphoinositide 3-kinase pathway. Science. 2002; 296:1655-7. 9. Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H, Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler KW, et al. High frequency of mutations of the PIK3CA gene in human cancers. Science. 2004; 304:554. ACKNOWLEDGMENTS 10. The Cancer Genome Atlas Research Network. Comprehensive molecular characterization of gastric adenocarcinoma. Nature. 2014; 513:202-9. We thank Anne M. 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https://openalex.org/W3123834268	https://discovery.ucl.ac.uk/10120681/1/ijms-22-01130-v2.pdf	English	null	Novel Biallelic Variants and Phenotypic Features in Patients with SLC38A8-Related Foveal Hypoplasia	International journal of molecular sciences	2,021	cc-by	9,519	Novel Biallelic Variants and Phenotypic Features in Patients with SLC38A8-Related Foveal Hypoplasia 1,2 , Vijay K. Tailor 1,3, Hwei Wuen Chan 1,2,4 , Maria Theodorou 1, Andrew R. Webster 1,2 osajee 1,2,5,6,* Elena R. Schiff 1,2 , Vijay K. Tailor 1,3, Hwei Wuen Chan 1,2,4 , Maria Theodorou 1, Andrew and Mariya Moosajee 1,2,5,6,* 1 Moorﬁelds Eye Hospital NHS Foundation Trust, London EC1V 2PD, UK; e.schiff@ucl.ac.uk (E.R.S.); vijay.tailor.09@ucl.ac.uk (V.K.T.); hwei_wuen_chan@nuhs.edu.sg (H.W.C.); mtheodorou@nhs.net (M.T.); Andrew.webster@ucl.ac.uk (A.R.W.) 1 Moorﬁelds Eye Hospital NHS Foundation Trust, London EC1V 2PD, UK; e.schiff@ucl.ac.uk (E.R.S.); vijay.tailor.09@ucl.ac.uk (V.K.T.); hwei_wuen_chan@nuhs.edu.sg (H.W.C.); mtheodorou@nhs.net (M.T.); Andrew.webster@ucl.ac.uk (A.R.W.) 1 Moorﬁelds Eye Hospital NHS Foundation Trust, London EC1V 2PD, UK; e.schiff@ucl.ac.uk (E.R.S.); vijay.tailor.09@ucl.ac.uk (V.K.T.); hwei_wuen_chan@nuhs.edu.sg (H.W.C.); mtheodorou@nhs.net (M. Andrew.webster@ucl.ac.uk (A.R.W.) 2 Institute of Ophthalmology, University College London, London EC1V 9EL, UK 3 Department of Experimental Psychology, University College London, London WC1H 0AP, UK 4 Department of Ophthalmology, National University Hospital, Singapore S118177, Singapore 5 Great Ormond Street Hospital for Children NHS Foundation Trust, London WC1N 3JH, UK 6 The Francis Crick Institute, London NW1 1AT, UK * Correspondence: m.moosajee@ucl.ac.uk 2 Institute of Ophthalmology, University College London, London EC1V 9EL, UK 3 Department of Experimental Psychology, University College London, London WC1H 0AP, UK 4 Department of Ophthalmology, National University Hospital, Singapore S118177, Singapore 5 Great Ormond Street Hospital for Children NHS Foundation Trust, London WC1N 3JH, UK 6 The Francis Crick Institute London NW1 1AT UK Institute of Ophthalmology, University College London, London EC1V 9EL, UK 3 Department of Experimental Psychology, University College London, London WC1H 0AP, UK 4 Department of Ophthalmology, National University Hospital, Singapore S118177, Singapore 5 Great Ormond Street Hospital for Children NHS Foundation Trust, London WC1N 3JH, UK 6 6 The Francis Crick Institute, London NW1 1AT, UK * Correspondence: m.moosajee@ucl.ac.uk Abstract: Biallelic pathogenic variants in solute carrier family 38 member 8, SLC38A8, cause a pan-ocular autosomal recessive condition known as foveal hypoplasia 2, FVH2, characterised by foveal hypoplasia, nystagmus and optic nerve chiasmal misrouting. Patients are often clinically diagnosed with ocular albinism, but foveal hypoplasia can occur in several other ocular disorders. Here we describe nine patients from seven families who had molecularly conﬁrmed biallelic recessive variants in SLC38A8 identiﬁed through whole genome sequencing or targeted gene panel testing. We identiﬁed four novel sequence variants (p.(Tyr88), p.(Trp145), p.(Glu233Gly) and c.632+1G>A). All patients presented with foveal hypoplasia, nystagmus and reduced visual acuity; however, one patient did not exhibit any signs of chiasmal misrouting, and three patients had features of anterior segment dysgenesis. Citation: Schiff, E.R.; Tailor, V.K.; Chan, H.W.; Theodorou, M.; Webster, A.R.; Moosajee, M. Novel Biallelic Variants and Phenotypic Features in Patients with SLC38A8-Related Foveal Hypoplasia. Int. J. Mol. Sci. 2021, 22, 1130. https://doi.org/ 10.3390/ijms22031130 Keywords: foveal hypoplasia; SLC38A8; nystagmus; chiasmal misrouting; anterior segment dysgenesis Academic Editor: Tomasz ˙Zarnowski Received: 31 December 2020 Accepted: 21 January 2021 Published: 24 January 2021 Citation: Schiff, E.R.; Tailor, V.K.; Chan, H.W.; Theodorou, M.; Webster, A.R.; Moosajee, M. Novel Biallelic Variants and Phenotypic Features in Patients with SLC38A8-Related Foveal Hypoplasia. Int. J. Mol. Sci. 2021, 22, 1130. https://doi.org/ 10.3390/ijms22031130 Novel Biallelic Variants and Phenotypic Features in Patients with SLC38A8-Related Foveal Hypoplasia We highlight these ﬁndings in the context of 30 other families reported to date. This study reinforces the importance of obtaining a molecular diagnosis in patients whose phenotype overlap with other inherited ocular conditions, in order to support genetic counselling, clinical prognosis and family planning. We expand the spectrum of SLC38A8 mutations which will be relevant for treatment through future genetic-based therapies. International Journal of Molecular Sciences International Journal of Molecular Sciences International Journal of Molecular Sciences International Journal of Molecular Sciences 1. Introduction The human fovea is located at the most central part of the macula, a 1.5 mm wide area most densely packed with cone photoreceptors. Its anatomy is distinct from other retinal regions, having a single layer of cones in the neurosensory retina with the inner nuclear and ganglion cell layer displaced radially outwards forming the foveal pit. Foveal hypoplasia (FVH) is the absence of this normal foveal depression, and the resultant continuation of all retinal layers and retinal vasculature through the foveal region. Grading systems describe the extent of FVH [1,2], and this can occur in isolation or as a feature of several ocular dis- orders including albinism, aniridia, retinopathy of prematurity, and microphthalmia. More recent associations, identiﬁed through high-resolution spectral domain optical coherence tomography (OCT) imaging, are achromatopsia, optic nerve hypoplasia, familial exudative vitreoretinopathy and Stickler syndrome [3]. For most—but not all—of these conditions, FVH is accompanied by nystagmus and decreased visual acuity, but this is dependent on the grade and severity. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional afﬁl- iations. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). When FVH occurs with optic nerve misrouting, it is usually an indication of albinism. Recessive mutations in the SLC38A8 (solute carrier family 38 member 8) gene on chro- mosome 16q23.3 have been shown, however, to cause FVH and optic nerve misrouting https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, 1130. https://doi.org/10.3390/ijms22031130 Int. J. Mol. Sci. 2021, 22, 1130 2 of 12 independent of the pigmentation defects associated with oculocutaneous albinism or oc- ular albinism [4,5]. Additionally known as foveal hypoplasia 2 with or without optic nerve misrouting and/or anterior segment dysgenesis (FVH2, OMIM #609218), SLC38A8 is a pan-ocular autosomal recessive condition, additionally labelled with the acronym FHONDA (foveal hypoplasia, optic nerve decussation and anterior segment dysgenesis without albinism) [6]. independent of the pigmentation defects associated with oculocutaneous albinism or oc- ular albinism [4,5]. Additionally known as foveal hypoplasia 2 with or without optic nerve misrouting and/or anterior segment dysgenesis (FVH2, OMIM #609218), SLC38A8 is a pan-ocular autosomal recessive condition, additionally labelled with the acronym FHONDA (foveal hypoplasia, optic nerve decussation and anterior segment dysgenesis without albinism) [6]. 1. Introduction SLC38A8 has 10 exons and encodes an amino acid transporter protein, which is one of 11 members of the SLC38 sodium-coupled neutral amino acid transporter (SNAT) family. The 435-amino acid protein SNAT8, has eleven transmembrane domains, an extracellular N-terminus and an intracellular C-terminal tail. The functional characterisation of SNAT8 showed that it is ubiquitously expressed in neurons and has a broad substrate proﬁle with high preference for transporting l-glutamine, l-alanine, l-arginine, l-histidine and l-aspartate. As a glutamine transporter, a key role for regulating levels of glutamate and gamma aminobutyric acid (GABA) in the brain has been suggested [7]. GABA/GABAergic dysregulation has a signiﬁcant impact on brain function from developmental to neurode- generative changes as seen in Alzheimer’s disease. It remains unknown whether this disruption in early development impacts on axonal projection, hence contributing to- wards chiasmal misrouting. Using human eye tissues, SLC38A8 was shown to be located throughout the neural retina, speciﬁcally in the inner and outer plexiform layers and the photoreceptor layer [4]. To date, 33 unique disease-causing mutations have been reported in 30 families of multiple ethnicities (Summarised in Table S1) [4,5,8–12], with an additional four only reported in ClinVar (total = 37). Almost half (n = 16) are missense mutations with the remainder being frameshift (n = 6), nonsense (n = 4), splice variant (n = 4) and deletion mutations (n = 7). In this study we detail the clinical and molecular ﬁndings in seven families with biallelic SLC38A8 mutations causing foveal hypoplasia, identiﬁed through whole genome sequencing (WGS), and through targeted gene panel testing, thus expanding the phenotypic and genotypic spectrum of SLC38A8-oculopathy. 2.1. Molecular Variant Findings Nine patients (6 females and 3 males) from 7 families with FVH and nystagmus were recruited into this study. Four families were of South Asian ethnicity with reported consanguinity, and three were Caucasian and non-consanguineous. The pedigrees of each family are shown in Figure 1 and the molecular ﬁndings summarised in Table 1. We identiﬁed 4 novel variants in SLC38A8 accounting for families 1–5; two nonsense (c.264C>G p.[Tyr88] and c.435G>A p.[Trp145]), one missense (c.698A>G, p.[Glu233Gly]) and one splice site (c.632+1G>A). A known missense heterozygous variant was found in family 6 (c.923C>G, p.[Thr308Ser]) together with a heterozygous deletion of exons 7 and 8 and a known homozygous missense was found in family 7 (c.848A>C, p.[Asp283Ala]). There are 37 reported mutations to date, and along with our novel variants these span all 10 exons (Figure 2A). The genotype of families 1, 3, 4 and 5 were reported in our previous study reporting the genetic outcomes of a large cohort of non-retinal developmental eye disease patients recruited into the 100,000 Genomes Project, but no detailed phenotype data was provided [13]. Variants were shown to be in trans in ﬁve of seven families where parents, all unaffected, were available for segregation analysis. Probands from families three and four were analysed as singletons. 3 of 12 of 13 Int. J. Mol. Sci. 2021, 22, 1130 nt. J. Mol. Sci. 2021, 22, x FOR PE Figure 1. Pedigrees of families with molecularly confirmed SLC38A8 biallelic variants, and corre- sponding retinal imaging. (A) Family 1 are consanguineous and (B) proband 1-1 with left ultra- widefield (UWF) colour fundus imaging; (C) Family 2 are consanguineous and (D) shows proband 2-1 right UWF colour fundus image, corresponding UWF fundus autofluorescence (FAF) with no defined fovea (usually depicted by a hypoautoautofluorescent spot in healthy individuals) and spectral-domain OCT (SD-OCT) demonstrating foveal hypoplasia through the centre of the right macula. Sibling 2-2 right UWF colour fundus and SD-OCT with foveal hypoplasia; (E) Family 4 are consanguineous and (F) proband 4-1 left UWF colour fundus with close up of the macula showing the concentric ring sign within the black dashed circle, which is seen in patients with foveal hypo- plasia; (G) Family 5 and (H) proband 5-1 showing left macula SD-OCT with foveal hypoplasia; (I) Family 6 and (J) proband 6-1 with left UWF colour fundus, UWF FAF with no foveal definition and SD-OCT with foveal hypoplasia. 2.1. Molecular Variant Findings Imaging was not available in family 3 and 7. Figure 1. Pedigrees of families with molecularly conﬁrmed SLC38A8 biallelic variants, and corre sponding retinal imaging. (A) Family 1 are consanguineous and (B) proband 1-1 with left ultra wideﬁeld (UWF) colour fundus imaging; (C) Family 2 are consanguineous and (D) shows proband 2-1 right UWF colour fundus image, corresponding UWF fundus autoﬂuorescence (FAF) with no deﬁned fovea (usually depicted by a hypoautoautoﬂuorescent spot in healthy individuals) and spectral-domain OCT (SD-OCT) demonstrating foveal hypoplasia through the centre of the right macula. Sibling 2-2 right UWF colour fundus and SD-OCT with foveal hypoplasia; (E) Family 4 are consanguineous and (F) proband 4-1 left UWF colour fundus with close up of the macula showing the concentric ring sign within the black dashed circle, which is seen in patients with foveal hypoplasia (G) Family 5 and (H) proband 5-1 showing left macula SD-OCT with foveal hypoplasia; (I) Family 6 and (J) proband 6-1 with left UWF colour fundus, UWF FAF with no foveal deﬁnition and SD-OCT with foveal hypoplasia. Imaging was not available in family 3 and 7. igure 1. Pedigrees of families with molecularly confirmed SLC38A8 biallelic variants, and corre- ponding retinal imaging. (A) Family 1 are consanguineous and (B) proband 1-1 with left ultra- widefield (UWF) colour fundus imaging; (C) Family 2 are consanguineous and (D) shows proband -1 right UWF colour fundus image, corresponding UWF fundus autofluorescence (FAF) with no efined fovea (usually depicted by a hypoautoautofluorescent spot in healthy individuals) and pectral-domain OCT (SD-OCT) demonstrating foveal hypoplasia through the centre of the right macula. Sibling 2-2 right UWF colour fundus and SD-OCT with foveal hypoplasia; (E) Family 4 are onsanguineous and (F) proband 4-1 left UWF colour fundus with close up of the macula showing he concentric ring sign within the black dashed circle, which is seen in patients with foveal hypo- lasia; (G) Family 5 and (H) proband 5-1 showing left macula SD-OCT with foveal hypoplasia; (I) amily 6 and (J) proband 6-1 with left UWF colour fundus, UWF FAF with no foveal definition and D-OCT with foveal hypoplasia. Imaging was not available in family 3 and 7. Figure 1. Pedigrees of families with molecularly conﬁrmed SLC38A8 biallelic variants, and corre- sponding retinal imaging. 2.1. Molecular Variant Findings It is also absent from gnomAD, though a different amino acid missense at this position, p.(Thr308Ala), has also been reported [9]. It is the only mutated amino acid in the eighth transmembrane domain. It is therefore likely that these missense mutations occur in locations that are critical for the normal function of the protein. A deletion spanning exons 7 and 8 was found in trans with c.923C>G, p.(Thr308Ser) in family 6. An exon 7-8 deletion has also been described in one patient of French origin in trans with c.697G>A, p.(Glu233Lys) (see Table S1). Table 1. SLC38A8 variants identiﬁed in our cohort using reference sequence NM_001080442.2. Abbrevia genome aggregation database; TM, transmembrane; HOM, homozygous; HET, heterozygous. Table 1. SLC38A8 variants identiﬁed in our cohort using reference sequence NM_001080442.2. Abbreviations: gnomAD, genome aggregation database; TM, transmembrane; HOM, homozygous; HET, heterozygous. Table 1. SLC38A8 variants identiﬁed in our cohort using reference sequence NM_001080442.2. Abbreviations: gnomAD, genome aggregation database; TM, transmembrane; HOM, homozygous; HET, heterozygous. Family ID Ethnicity Consanguinity Variants Exon Consequence Protein Domain Zygosity Reported gnomAD 1 (26320) South Asian Bangladesh Yes c.264C>G p.(Tyr88) 2 Nonsense Intracellular between TM domains 1 and 2 HOM Novel 0.0002614 in South Asians only 2 (23089) South Asian Indian Yes c.264C>G p.(Tyr88) 2 Nonsense Intracellular between TM domains 1 and 2 HOM Novel 0.0002614 in South Asians only 3 (26350) South Asian Pakistan Yes c.264C>G p.(Tyr88) 2 Nonsense Intracellular between TM domains 1 and 2 HOM Novel 0.0002614 in South Asians only 4 (26364) South Asian Sri-Lanka Yes c.698A>G p.Glu233Gly 6 Missense TM6 HOM Novel Absent 5 (26237) Caucasian British No c.435G>A p.Trp145 3 Nonsense Extracellular between TM domains 3 and 4 HET Novel Absent c.632+1G>A IVS4 Splice donor Extracellular between TM domains 5 and 6 HET Novel Absent 6 (16812) Caucasian Spanish- British No c.923C>G p.Thr308Ser Del Ex7-8 7 Missense splice site impact Deletion TM8 TM7-TM10 HET HET [9] [9] Absent Absent 7 (28327) Caucasian Ashkenazi Jewish No c.848A>C p.(Asp283Ala) 7 Missense Extracellular between TM domains 7 and 8 HOM [8,9] 0.0002653 in gnomAD 0.006178 in AJs The homozygous missense variant c.848A>C, p.(Asp283Ala) found in family 7 has previously been reported in both a homozygous [8,14] and compound heterozygous state with the pathogenic variant c.527C>G, p.(Thr176Arg) [9]. 2.1. Molecular Variant Findings (A) Family 1 are consanguineous and (B) proband 1-1 with left ultra- wideﬁeld (UWF) colour fundus imaging; (C) Family 2 are consanguineous and (D) shows proband 2-1 right UWF colour fundus image, corresponding UWF fundus autoﬂuorescence (FAF) with no deﬁned fovea (usually depicted by a hypoautoautoﬂuorescent spot in healthy individuals) and spectral-domain OCT (SD-OCT) demonstrating foveal hypoplasia through the centre of the right macula. Sibling 2-2 right UWF colour fundus and SD-OCT with foveal hypoplasia; (E) Family 4 are consanguineous and (F) proband 4-1 left UWF colour fundus with close up of the macula showing the concentric ring sign within the black dashed circle, which is seen in patients with foveal hypoplasia; (G) Family 5 and (H) proband 5-1 showing left macula SD-OCT with foveal hypoplasia; (I) Family 6 and (J) proband 6-1 with left UWF colour fundus, UWF FAF with no foveal deﬁnition and SD-OCT with foveal hypoplasia. Imaging was not available in family 3 and 7. The nonsense mutation c.264C>G, p.(Tyr88), observed homozygously in families 1, 2 and 3, lies between transmembrane domains 2–3, in a region deplete of reported variants (Figure 2A). It is likely that transcripts encoding nonsense and frameshift variants will be removed by nonsense-mediated decay or alternatively result in a truncated protein. In family 5, the novel nonsense mutation c.435G>A, p.(Trp145) lies between transmembrane domains 3-4, in proximity to another nonsense mutation, p.(Gln149Ter), listed in ClinVar (rs14689932). This may be a region susceptible to protein function disruption. The novel Int. J. Mol. Sci. 2021, 22, 1130 4 of 12 splice variant c.632+1G>A observed in trans with p.(Trp145) in family 5 alters the con- served canonical donor splice site at the exon4-intron4 junction, and a different splice mu- tation at this site, c.632+2T>G, was recently described [12]. The novel missense c.698A>G, p.(Glu233Gly) in family 4 is predicted to be probably damaging/damaging/disease-causing by Polyphen2, SIFT and Mutation Taster, respectively. It involves a highly conserved residue within a conserved block (Figure 2B) and is absent from gnomAD. It occurs in the sixth transmembrane domain which harbours 5 other missense variants and is the most frequently affected domain. The previously reported [9] missense c.923C>G, p.(Thr308Ser) heterozygous variant in family 6 is similarly predicted to be likely pathogenic and is also positioned in a highly conserved amino acid residue block (Figure 2B). 2.1. Molecular Variant Findings It occurs in the extracellular do- main between the 7th and 8th transmembrane domains and structural modelling predicted a destabilizing effect on the protein by altering the electrostatic potential within the channel pore [8]. Family 7 were of Ashkenazi Jewish descent and this variant is relatively common in this population (0.006178 compared to 0.0002653 in gnomAD overall). The homozygous missense variant c.848A>C, p.(Asp283Ala) found in family 7 has previously been reported in both a homozygous [8,14] and compound heterozygous state with the pathogenic variant c.527C>G, p.(Thr176Arg) [9]. It occurs in the extracellular do- main between the 7th and 8th transmembrane domains and structural modelling predicted a destabilizing effect on the protein by altering the electrostatic potential within the channel pore [8]. Family 7 were of Ashkenazi Jewish descent and this variant is relatively common in this population (0.006178 compared to 0.0002653 in gnomAD overall). The homozygous missense variant c.848A>C, p.(Asp283Ala) found in family 7 has previously been reported in both a homozygous [8,14] and compound heterozygous state with the pathogenic variant c.527C>G, p.(Thr176Arg) [9]. It occurs in the extracellular do- main between the 7th and 8th transmembrane domains and structural modelling predicted a destabilizing effect on the protein by altering the electrostatic potential within the channel pore [8]. Family 7 were of Ashkenazi Jewish descent and this variant is relatively common in this population (0.006178 compared to 0.0002653 in gnomAD overall). The homozygous missense variant c.848A>C, p.(Asp283Ala) found in family 7 has previously been reported in both a homozygous [8,14] and compound heterozygous state with the pathogenic variant c.527C>G, p.(Thr176Arg) [9]. It occurs in the extracellular do- main between the 7th and 8th transmembrane domains and structural modelling predicted a destabilizing effect on the protein by altering the electrostatic potential within the channel pore [8]. Family 7 were of Ashkenazi Jewish descent and this variant is relatively common in this population (0.006178 compared to 0.0002653 in gnomAD overall). p y p yg p yg with the pathogenic variant c.527C>G, p.(Thr176Arg) [9]. It occurs in the extracellular do- main between the 7th and 8th transmembrane domains and structural modelling predicted a destabilizing effect on the protein by altering the electrostatic potential within the channel pore [8]. Family 7 were of Ashkenazi Jewish descent and this variant is relatively common in this population (0.006178 compared to 0.0002653 in gnomAD overall). 5 of 12 nomAD 6178 in Int. J. Mol. Sci. 2.1. Molecular Variant Findings 2021, 22, 1130 7 (28327) Caucasian Ashkenazi Figure 2. SLC38A8 variants and corresponding protein alignment of two missense mutations. (A) Schematic representation of SLC38A8 gene transcript showing localisation of mutations reported to date (Table S1) above the (B) Alignment (A) SLC38A8 gene Del Ex7-8 Figure 2. SLC38A8 variants and corresponding protein alignment of two missense mutations. (A) Schematic representation of SLC38A8 gene transcript showing localisation of mutations reported to date (Table S1) above the transcript and mutations described in this study below, novel mutations in bold. Red vertical lines represent exon boundaries; shaded areas represent the eleven transmembrane domains, TM1-TM11; (B) Multiple organism sequence alignment of regions spanning the p.Glu233Gly and p.Thr308Ser amino acids of the SLC38A8 protein, showing their strong evolutionary conservation (highly conserved in darker grey moderately conserved in lighter grey). Alignment was performed using https://www.uniprot.org/ align/. Protein sequences used for alignment are S38A8 in the human, H2RA07 in the chimpanzee, F6Q2Q9 in the rhesus monkey, M3WFB0 in the cat, S38A8 in the mouse, F1NJ98 in the chicken, S38A8 in the frog and F1QK49 in the zebraﬁsh. (A) SLC38A8 gene Del Ex7-8 (A) SLC38A8 gene (B) Alignment Figure 2. SLC38A8 variants and corresponding protein alignment of two missense mutations. (A) Schematic representation of SLC38A8 gene transcript showing localisation of mutations reported to date (Table S1) above the Figure 2. SLC38A8 variants and corresponding protein alignment of two missense mutations. (A) Schematic representation of SLC38A8 gene transcript showing localisation of mutations reported to date (Table S1) above the transcript and mutations described in this study below, novel mutations in bold. Red vertical lines represent exon boundaries; shaded areas represent the eleven transmembrane domains, TM1-TM11; (B) Multiple organism sequence alignment of regions spanning the p.Glu233Gly and p.Thr308Ser amino acids of the SLC38A8 protein, showing their strong evolutionary conservation (highly conserved in darker grey moderately conserved in lighter grey). Alignment was performed using https://www.uniprot.org/ align/. Protein sequences used for alignment are S38A8 in the human, H2RA07 in the chimpanzee, F6Q2Q9 in the rhesus monkey, M3WFB0 in the cat, S38A8 in the mouse, F1NJ98 in the chicken, S38A8 in the frog and F1QK49 in the zebraﬁsh. 2.2. Clinical Findings The ophthalmic and ocular ﬁndings of patients with SLC38A8 variants are summarised in Table 2 and Figure 1. The mean ± SD BCVA was 0.73 ± 0.15 LogMAR. All nine patients had FVH and eight out of nine patients demonstrated a horizontal nystagmus waveform type (7 with pendular nystagmus and 1 had a jerk with extended foveation), and one patient had a rotary nystagmus. FVH was recorded through either ophthalmic examination or retinal imaging using fundus autoﬂuorescence (FAF) or SD-OCT. FAF revealed an absence of the foveal hypoautoﬂuorescent spot, hence no deﬁned fovea as shown in Figure 1D and J. SD-OCT was challenging due to lack of patient ﬁxation resulting from the FVH and nystagmus, however using the best single line scans, grade 4 FVH was seen in six families, graded according to Thomas et al. [8] Grade 4 FVH includes absence of the following features: (i) plexiform layers, (ii) foveal pit, (iii) outer segment lengthening and (iv) outer nuclear layer widening (Figure 1D,F,H,J). Close inspection of the UWF colour fundus images revealed the concentric macular ring sign in four patients; 1-1, 2-1, 4-1 (shown in Figure 1F) and 6-1. Its appearance is reliant on the quality of the images, which as with OCT is confounded by age and ﬁxation. This is a documented sign of FVH which has been postulated to arise from the radially symmetric orientation of the axon photoreceptors (Henle ﬁbre layer) and possibly also the retinal nerve ﬁbre layer surrounding the fovea [15,16]. Int. J. Mol. Sci. 2021, 22, 1130 6 of 12 Table 2. Clinical ﬁndings in patients with SLC38A8 variants. Best corrected visual acuity (BCVA) recorded in logMAR (OD, right eye, and OS, left eye). 2.2. Clinical Findings Six patients who underwent electrophysiology with pattern appearance visual evoked potentials (VEP) had evidence of intracranial misrouting with contralateral predominance (crossed asymmetry). Three patients (2-2, 4-1 and 6-1) had variable degrees of anterior segment dysgenesis, two had bilateral posterior embryotoxon, 4-1 also had bilateral iris adhesions to the cornea and bilateral blue dot cataracts, and 6-1 had bilateral shallow an- terior chamber. In family 2 of Indian Gujarati background both the affected siblings were initially In family 2, of Indian Gujarati background, both the affected siblings were initially reported as having paler skin than their three unaffected siblings and parents, however, over time their skin pigmentation was similar to that of the rest of the family. Their hair and eyes were no different to that of their parents. In family 5, of Caucasian ethnicity, it was noted that the two affected children had fair hair and complexion, but this was in common with both their parents and many fair-haired and blue-eyed relatives. The remaining patients had normal hair and skin pigment compared to relatives and no ocular hypopigmentation. In family 2, of Indian Gujarati background, both the affected siblings were initially reported as having paler skin than their three unaffected siblings and parents, however, over time their skin pigmentation was similar to that of the rest of the family. Their hair and eyes were no different to that of their parents. In family 5, of Caucasian ethnicity, it was noted that the two affected children had fair hair and complexion, but this was in common with both their parents and many fair-haired and blue-eyed relatives. The re- maining patients had normal hair and skin pigment compared to relatives and no ocular hypopigmentation. All previously documented cases in the literature (30 families) were analysed for the All previously documented cases in the literature (30 families) were analysed for the prevalence of SLC38A8 phenotypic features including our cohort (seven families). Of the 58 affected individuals in 37 families, 100% (n = 58) had foveal hypoplasia and nystagmus, while 97% (29 out of the 30 who underwent VEP testing) showed chiasmal misrouting. Anterior segment dysgenesis was documented in 22% (13/58) of patients with 21% (n = 12) having posterior embryotoxon and 5% (n = 3) showing Axenfeld anomaly (deﬁned as posterior embryotoxon with peripheral anterior iris adhesions). 2.2. Clinical Findings Family ID/Patient Ethnicity Age (Years) Gender BCVA OD BCVA OS Refraction Strabismus Nystagmus Anterior Segment Foveal Hypoplasia Electrodiagnostic Testing 1-1 South Asian Bangladesh 3 Male Objects to occlusion Objects to occlusion R: +3.50/−2.50 × 180 L: +3.50/−3.50 × 180 Left Exotropia Pendular horizontal Normal Present No evidence of chiasmal misrouting 2-1 South Asian Indian 32 Female 0.6 0.6 R: −1.75/−3.50 × 180 L: −7.00/−2.50 × 170 Left Esotropia Pendular horizontal Normal Grade 4 Evidence of chiasmal misrouting 2-2 16 Male 0.82 0.78 R: −0.75/−3.50 × 10 L: +1.50/−4.00 × 175 Right Exotropia Pendular horizontal Posterior embryotoxon Grade 4 Evidence of chiasmal misrouting 3-1 South Asian Pakistan 21 Female 0.7 0.9 R: +0.75/−2.50 × 30 L: +0.75/−2.50 × 75 Left Exotropia Pendular horizontal Normal Present Not undertaken 4-1 South Asian Sri-Lanka 39 Female 0.6 0.6 Not available Left Exotropia Pendular horizontal Bilateral Posterior embryotoxon Bilateral peripheral iris adhesions to the cornea Bilateral blue dot cataract Grade 4 Evidence of chiasmal misrouting 5-1 Caucasian British 5 Female 0.6 0.6 R: +3.50DS L: +3.50DS No deviation Jerk with extended foveation Normal Grade 4 Evidence of chiasmal misrouting 5-2 2 Male Objects to occlusion Objects to occlusion R: +5.00/−0.75 × 180 L: +5.00/−0.75 × 180 No deviation Pendular horizontal Normal Present Not undertaken 6-1 Caucasian Spanish- British 36 Female 0.9 1 Not available Left Esotropia Rotary Bilateral shallow anterior chamber Grade 4 Evidence of chiasmal misrouting 7-1 Caucasian Ashkenazi Jewish 16 Female 0.42 0.4 R: +2.50/−1.50 × 165 L: +1.50/−1.50 × 180 Right Exotropia Pendular horizontal Peripheral iris transillumination Grade 4 Evidence of chiasmal misrouting Table 2. Clinical ﬁndings in patients with SLC38A8 variants. Best corrected visual acuity (BCVA) re ariants. Best corrected visual acuity (BCVA) recorded in logMAR (OD, right eye, and OS, left eye). Int. J. Mol. Sci. 2021, 22, 1130 7 of 12 Seven patients had a strabismus with exotropia in ﬁve patients and esotropia in two. Six patients who underwent electrophysiology with pattern appearance visual evoked potentials (VEP) had evidence of intracranial misrouting with contralateral predominance (crossed asymmetry). Three patients (2-2, 4-1 and 6-1) had variable degrees of anterior segment dysgenesis, two had bilateral posterior embryotoxon, 4-1 also had bilateral iris adhesions to the cornea and bilateral blue dot cataracts, and 6-1 had bilateral shallow anterior chamber. Seve patie ts ad a st abis us wit e ot opia i ive patie ts a d esot opia i two. 2.2. Clinical Findings Strabismus was observed in 54% (20/37) and iris transillumination in 7% (4/58) (see Figure 3 and Table S2). All previously documented cases in the literature (30 families) were analysed for the prevalence of SLC38A8 phenotypic features including our cohort (seven families). Of the 58 affected individuals in 37 families, 100% (n = 58) had foveal hypoplasia and nystagmus, while 97% (29 out of the 30 who underwent VEP testing) showed chiasmal misrouting. Anterior segment dysgenesis was documented in 22% (13/58) of patients with 21% (n = 12) having posterior embryotoxon and 5% (n = 3) showing Axenfeld anomaly (defined as posterior embryotoxon with peripheral anterior iris adhesions). Strabismus was observed in 54% (20/37) and iris transillumination in 7% (4/58) (see Figure 3 and Table S2). Figure 3. Bubble plot representing the relative frequency of phenotypic features in 58 reported cases (including our cohort) in 37 families. Anterior segment dysgenesis encompasses all recorded fea- tures including posterior embryotoxon, Axenfeld anomaly, blue-dot cataract, shallow anterior seg- ments and iris transillumination. Foveal hypoplasia Nystagmus Chiasmal misrouting ASD Strabismus Iris transillumination 100% 100% 22% 54% 97% 7% SLC38A8 phenotype Figure 3. Bubble plot representing the relative frequency of phenotypic features in 58 reported cases (including our cohort) in 37 families. Anterior segment dysgenesis encompasses all recorded features including posterior embryotoxon, Axenfeld anomaly, blue-dot cataract, shallow anterior segments and iris transillumination. Foveal hypoplasia Nystagmus Chiasmal misrouting ASD Strabismus Iris transillumination 100% 100% 22% 54% 97% 7% SLC38A8 phenotype Foveal hypoplasia Chiasmal misrouting Chiasmal misrouting Strabismus Figure 3. Bubble plot representing the relative frequency of phenotypic features in 58 reported cases including our cohort) in 37 families. Anterior segment dysgenesis encompasses all recorded fea- ures including posterior embryotoxon, Axenfeld anomaly, blue-dot cataract, shallow anterior seg- ments and iris transillumination. Figure 3. Bubble plot representing the relative frequency of phenotypic features in 58 reported cases (including our cohort) in 37 families. Anterior segment dysgenesis encompasses all recorded features including posterior embryotoxon, Axenfeld anomaly, blue-dot cataract, shallow anterior segments and iris transillumination. 3. Discussion 3. Discussion i u io This study reports four novel variants in SLC38A8 amongst 7 families, with the asso- ciated clinical features, and compares these to previously described molecularly con- firmed cases. The homozygous nonsense mutation c.264C>G, p.(Tyr88) was found in three unrelated families from different regions of South Asia. This variant was rare in gnomAD, but found with an allele frequency of 0.0002614 only in the South Asian popu- lation suggesting that this represents a possible founder mutation This study reports four novel variants in SLC38A8 amongst 7 families, with the associ- ated clinical features, and compares these to previously described molecularly conﬁrmed cases. The homozygous nonsense mutation c.264C>G, p.(Tyr88*) was found in three unre- lated families from different regions of South Asia. This variant was rare in gnomAD, but found with an allele frequency of 0.0002614 only in the South Asian population suggesting that this represents a possible founder mutation. ation suggesting that this represents a possible founder mutation. Foveal hypoplasia and nystagmus were present in all patients, with 83.3% (5/6 pa- tients) showing evidence of intracranial misrouting. Prior to 2013, visual pathway develop- mental defects of foveal hypoplasia and intracranial misrouting were considered pathog- Int. J. Mol. Sci. 2021, 22, 1130 8 of 12 nomonic of albinism, and the associated defects in the melanin-biosynthesis pathway [4,6]. It has since been suggested that SLC38A8-related features occur in a melanin-independent manner with no effect on pigmentation. In our cohort two families were described as having paler skin (Family 2 and 5) and fair hair (Family 5), hence were clinically diagnosed with oculocutaneous/ocular albinism prior to genetic testing. Over time, the affected individuals in both families reached similar levels of pigmentation to their parents and siblings. In all the other reported families in the literature, there were no signs of cutaneous hypopigmentation. In addition, more variability in grade of foveal hypoplasia is seen in patients with albinism and PAX6 variants compared to SLC38A8 patients, who have a more severe retinal phenotype indicating an earlier arrest in foveal development [12]. Anterior segment dysgenesis (ASD) encompasses a spectrum of clinical features originating from maldevelopment of the anterior segment, which can involve all or any part of the cornea, anterior chamber, trabecular meshwork, iris, ciliary body, and lens. Our cohort, together with published cases report ASD features ranging from posterior embryotoxon alone to Axenfeld anomaly, blue-dot cataract and shallow anterior chambers. 3. Discussion 3. Discussion The prevalence of posterior embryotoxon in the younger (≤20 years of age) population has been reported to be 22.5% [17]. Analysis of all SLC38A8 reported cases reveals a similar prevalence of 22% (13/58), but in our cohort alone, this increases to 33% (3/9). The SLC38A8 variants in all patients with ASD were not localised to the same region/domain, so there was no genotype-phenotype correlation. Hence, it remains unclear if this is a variable feature of the disorder or a coincidental ﬁnding. Human gene expression datasets show that SLC38A8 transcript (ENST00000299709.7) is differentially expressed across the eye including the foetal retina (7.75 log2(TPM + 1)), foetal RPE (5.56 log2(TPM + 1)), cornea endothelium (1.03 log2(TPM + 1)) and stem cell derived lens (2.92 log2(TPM + 1)) [18]. This pan-ocular expression is similar to PAX6, which also plays a signiﬁcant role in foveal and anterior segment development; hence, further deeper phenotyping may be required with the use of anterior segment OCT to carefully capture the morphology of these affected eyes. g y p p gy y Isolated foveal hypoplasia without optic nerve misrouting or anterior segment dysge- nesis was found in one family (family 1). Differential diagnoses includes autosomal domi- nant foveal hypoplasia-1 (FVH1, OMIM #136520), also referred to as foveal hypoplasia-1 with or without anterior segment anomalies and/or cataract, which is caused by het- erozygous missense variants in the PAX6 gene on chromosome 11p13 [19,20]. Hemizygous mutations in GPR143 on chromosome Xp22, commonly cause ocular albinism type I (OMIM 300500); however, in a smaller number of cases, it has been reported to cause isolated con- genital nystagmus-6 (OMIM 300814). In a six-generation Chinese family with a c.266C>T, p.(Ser89Phe) variant, affected members displayed nystagmus and foveal hypoplasia, with- out any typical signs of ocular albinism such as iris transillumination defects, fundus hypopigmentation and intracranial misrouting [21]. In a Korean family with a GPR143 c.623C>A, p.(Ala208Glu) variant, one affected male displayed typical ocular albinism signs with severe hypopigmentation with clearly visible choroid vessels in the entire retina and eye movement recording showed a horizontal, conjugate pendular nystagmus. His affected sibling had isolated foveal hypoplasia with no nystagmus nor iris or fundus hypopigmen- tation [22]. Electrophysiology was not reported in this study. Care should be taken as iris and fundus hypopigmentation may not always be apparent in Asians. 3. Discussion 3. Discussion All but one of the patients in this study were solved through whole genome sequenc- ing facilitated by the 100,000 Genomes Project, highlighting its utility to determine a molecular diagnosis, especially where the phenotypic overlap with albinism, aniridia and other ocular conditions can lead to potential misdiagnosis. Recognising the phenotype of SLC38A8-oculopathy (and pattern of inheritance) will enable the clinical genetics team to apply supporting evidence for pathogenicity (PP4) when required to conﬁrm a molecular diagnosis of a SLC38A8 variant of uncertain signiﬁcance [23]. In addition, reaching an accurate diagnosis is especially important for informed genetic counselling and family planning advice, given the different inheritance patterns related to the other disease phe- nocopies. There are emerging gene-based applications for treating albinism [24] such as Int. J. Mol. Sci. 2021, 22, 1130 9 of 12 9 of 12 adeno-associated virus (AAV)-based Oa1 gene therapy which has been demonstrated in knockout mice models for ocular albinism [25] and with Tyr oculocutaneous albinism type 1 [26]. Successful CRISPR/Cas9 gene editing of Tyr has also been demonstrated for albinism in rabbits [27]. These pigment targeted gene-based therapies will be relevant for future therapies of patients with albinism, for example, through the restoration of tyrosinase production, and will depend on accurate genetic diagnosis. y p p g g SLC38A8-oculopathy may well be under diagnosed because of its subtle presentation, easily confused with albinism. Current molecular diagnostic rates for albinism and nystag- mus using WGS are approximately 28% [13]. Until recently, SLC38A8 was not included in the albinism and nystagmus targeted gene panels. As found with patients presenting with nystagmus [28], more comprehensive gene panels offered as an initial assessment will limit such diagnostic delays. Detailed phenotyping of patients will also help to characterise the pan-ocular features associated with this condition. 4.1. Patients and Genetic Analysis All patients attended the ocular genetics service at MEH. Patients and relatives gave written informed consent for participation in this study through either the Genetic Study of Inherited Eye Disease (REC reference 12/LO/0141) or Genomics England 100,000 Genomes project (REC reference 14/EE/1112). Families 1-6 comprising of probands and one or two unaffected parents where available) had undergone WGS analysis through participation in the 100,000 Genomes Project [29]. Detailed by Taylor et al. [30], genomic DNA was processed using an Illumina TruSeq DNA PCR-Free Sample Preparation kit (Illumina Inc., San Diego, CA, USA) and sequenced using an Illumina HiSeq X Ten high-throughput sequencing platform, generating minimum coverage of 15 X for >97% of the callable auto- somal genome. Readings were aligned to either build GRCh37 or GRCH38 of the human genome using an Isaac aligner (Illumina Inc., San Diego, CA, USA). Single-nucleotide variants (SNVs) and indels (insertions or deletions) were identiﬁed using Platypus soft- ware (version 0.8.1; and annotated using Cellbase software (https://github.com/opencb/ cellbase). Variant ﬁltering was performed using minor allele frequency (MAF) < 0.001 in publicly available and in-house data sets, predicted protein effect, and familial seg- regation. Surviving variants were prioritized using the Retinal disorders version 2.120 (https://panelapp.genomicsengland.co.uk/panels/307/) and the Albinism or congenital nystagmus version 1.10 (https://panelapp.genomicsengland.co.uk/panels/511/) virtual gene panels. g p The proband from family 7 underwent targeted albinism and nystagmus gene panel testing through the Rare & Inherited Disease Genomic Laboratory at Great Ormond Street Hospital (London, UK). Mutation screening was carried out by next generation sequencing with library preparation using the Agilent focused clinical exome +1 kit followed by sequencing on the Illumina platforms. Data were analysed using an in-house pipeline and virtual gene panels including SLC38A8, with all mutations conﬁrmed by Sanger sequencing. k l h f h l d h d g p The proband from family 7 underwent targeted albinism and nystagmus gene panel testing through the Rare & Inherited Disease Genomic Laboratory at Great Ormond Street Hospital (London, UK). Mutation screening was carried out by next generation sequencing with library preparation using the Agilent focused clinical exome +1 kit followed by sequencing on the Illumina platforms. Data were analysed using an in-house pipeline and virtual gene panels including SLC38A8, with all mutations conﬁrmed by Sanger sequencing. 4.2. Clinical Assessment 4.2. Clinical Assessment Ophthalmic examination included full orthoptic assessment, refraction, best corrected visual acuity (VA) measured using LogMAR or Cardiff cards for preverbal children up to 36 months of age; slit lamp examination, and fundus examination were recorded with anterior segment and ultra-wideﬁeld (UWF) fundus colour imaging using the Haag-Streit slit lamp camera (Haag-Streit Holdings AG, Köniz, Switzerland) and Optos® California (Optos plc), respectively. Investigations included spectral domain optical coherence tomography (SD-OCT), fundus autoﬂuorescence, eye movement recordings (EMR) to assess nystag- mus waveforms, electrophysiology to detect chiasmal misrouting of retinal ganglion cells through visual evoked potentials (VEP) against ISCEV standards, and either multichannel ﬂash VEP alone (for younger paediatric patients) or a combination of both ﬂash and pattern VEP. Due to the nystagmus and foveal hypoplasia seen in all patients, retinal imaging was challenging due to ﬁxation difﬁculties and compliance in young children. Multiple line scan SD-OCT was attempted, but reverted to multiple single line scans where not possible to capture the foveal pit. Supplementary Materials: The following are available online at https://www.mdpi.com/1422-0 067/22/3/1130/s1, Table S1: Published variants in SLC38A8, Table S2: Percentage of phenotypic features seen in previously documented cases. Author Contributions: Conceptualization, M.M. and E.R.S.; clinical data, M.M., A.R.W., V.K.T., H.W.C. and M.T.; analysis and interpretation, M.M., E.R.S., V.K.T. and A.R.W.; writing—original draft preparation, E.R.S., V.K.T.; writing—review and editing, M.M., A.R.W. All authors have read and agreed to the published version of the manuscript. Funding: The research was supported by the Wellcome Trust (Grant No. 205174/Z/16/Z). We gratefully acknowledge the support of the National Institute for Health Research (NIHR) Biomedical Research Centre based at Moorﬁelds Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology. The views expressed are those of the authors and not the funding organisations. Institutional Review Board Statement: This study had relevant local and national research ethics committee approvals (Moorﬁelds Eye Hospital NHS Foundation Trust [MEH] and the Northwest London Research Ethics Committee), and adhered to the tenets of the Declaration of Helsinki. Patients and relatives gave written informed consent for genetic testing through either the Genetic Study of Inherited Eye Disease (REC reference 12/LO/0141) or Genomics England 100,000 Genomes project (REC reference 14/EE/1112). Informed Consent Statement: Informed consent was obtained from all subjects involved in the study. 4.1. Patients and Genetic Analysis Likely pathogenicity of the novel missense variant was assessed using the predictive algorithms PolyPhen-2 [31], SIFT [32] and Mutation Taster [33].The genetic results were reviewed by a multidisciplinary team (including molecular biologists, clinical geneticists, as well as the ophthalmology specialist managing the family), to conﬁrm variant pathogenic- The proband from family 7 underwent targeted albinism and nystagmus gene panel testing through the Rare & Inherited Disease Genomic Laboratory at Great Ormond Street Hospital (London, UK). Mutation screening was carried out by next generation sequencing with library preparation using the Agilent focused clinical exome +1 kit followed by sequencing on the Illumina platforms. Data were analysed using an in-house pipeline and virtual gene panels including SLC38A8, with all mutations conﬁrmed by Sanger sequencing. Likely pathogenicity of the novel missense variant was assessed using the predictive algorithms PolyPhen-2 [31], SIFT [32] and Mutation Taster [33].The genetic results were reviewed by a multidisciplinary team (including molecular biologists, clinical geneticists, as well as the ophthalmology specialist managing the family), to conﬁrm variant pathogenic- ity, prevalence in publicly available genome databases, the clinical phenotype and mode of inheritance, before the molecular diagnosis was established. The datasets (variants) gener- ated for this study were submitted to ClinVar (https://www.ncbi.nlm.nih.gov/clinvar/). We describe nine patients from seven unrelated families who were clinically diagnosed with bilateral foveal hypoplasia and nystagmus, and molecularly conﬁrmed biallelic mutations in SLC38A8. Likely pathogenicity of the novel missense variant was assessed using the predictive algorithms PolyPhen-2 [31], SIFT [32] and Mutation Taster [33].The genetic results were reviewed by a multidisciplinary team (including molecular biologists, clinical geneticists, as well as the ophthalmology specialist managing the family), to conﬁrm variant pathogenic- ity, prevalence in publicly available genome databases, the clinical phenotype and mode of inheritance, before the molecular diagnosis was established. The datasets (variants) gener- ated for this study were submitted to ClinVar (https://www.ncbi.nlm.nih.gov/clinvar/). We describe nine patients from seven unrelated families who were clinically diagnosed with bilateral foveal hypoplasia and nystagmus, and molecularly conﬁrmed biallelic mutations in SLC38A8. 10 of 12 Int. J. Mol. Sci. 2021, 22, 1130 10 of 12 4.2. Clinical Assessment Data Availability Statement: The variants reported in this paper have been deposited into the ClinVar database (https://www.ncbi.nlm.nih.gov/clinvar/) at the National Center for Biotech- nology Information under accession numbers RCV001270481, SCV001450776, SCV001450777 and SCV001450778. Acknowledgments: We are grateful to patients and relatives for taking part in this research. This research was made possible through access to the data and ﬁndings generated by the 100,000 Genomes Project; http://www.genomicsengland.co.uk. Conﬂicts of Interest: The authors declare no conﬂict of interest. Conﬂicts of Interest: The authors declare no conﬂict of interest. 1. Thomas, M.G.; Kumar, A.; Mohammad, S.; Proudlock, F.A.; Engle, E.C.; Andrews, C.; Chan, W.M.; Thomas, S.; Gottlob, I. Structural grading of foveal hypoplasia using spectral-domain optical coherence tomography: A predictor of visual acuity? Ophthalmology 2011, 118, 1653–1660. [CrossRef] [PubMed] 2. 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https://openalex.org/W4362594316	https://link.springer.com/content/pdf/10.1007/s11060-023-04333-3.pdf	English	null	Sonodynamic therapy and Magnetic Resonance-guided Focused Ultrasound: new therapeutic strategy in Glioblastoma	Research Square (Research Square)	2,023	cc-by	12,690	1 Department of Biomedicine Neurosciences and Advanced Diagnostics, School of Medicine, Neurosurgical Clinic, AOUP “Paolo Giaccone”, Post Graduate Residency Program in NeurologiSurgery, University of Palermo, 90127 Palermo, Italy Abstract Glioblastoma (GB) is one of the most aggressive and difficult-to-treat brain tumors, with a poor prognosis and limited treat- ment options. In recent years, sonodynamic therapy (SDT) and magnetic resonance focused ultrasound (MRgFUS) have emerged as promising approaches for the treatment of GB. SDT uses ultrasound waves in combination with a sonosensitizer to selectively damage cancer cells, while MRgFUS delivers high-intensity ultrasound waves to precisely target tumor tis- sue and disrupt the blood–brain barrier to enhance drug delivery. In this review, we explore the potential of SDT as a novel therapeutic strategy for GB. We discuss the principles of SDT, its mechanisms of action, and the preclinical and clinical studies that have investigated its use in Gliomas. We also highlight the challenges, the limitations, and the future perspectives of SDT. Overall, SDT and MRgFUS hold promise as novel and potentially complementary treatment modalities for GB. Further research is needed to optimize their parameters and determine their safety and efficacy in humans, but their potential for selective and targeted tumor destruction makes them an exciting area of investigation in the field of brain cancer therapy. Keywords Foscused Ultrasound · Gliomas · Glioblastoma · Sonodynamic Therapy · MRgFUS * Lapo Bonosi lapo.bonosi@gmail.com Sonodynamic therapy and magnetic resonance‑guided focused ultrasound: new therapeutic strategy in glioblastoma Received: 2 April 2023 / Accepted: 3 May 2023 / Published online: 14 May 2023 © The Author(s) 2023 2 IRCCS Centro Neurolesi Bonino-Pulejo, Messina, Italy RESEARCH RESEARCH RESEARCH Journal of Neuro-Oncology (2023) 163:219–238 https://doi.org/10.1007/s11060-023-04333-3 Journal of Neuro-Oncology (2023) 163:219–238 https://doi.org/10.1007/s11060-023-04333-3 Introduction management of GB has not changed since Roger Stupp published his work [4]. Gliomas represent about 25% of all primary brain tumors, encompassing malignant and not malignant subtypes [1]. IDH-wildtype glioblastoma (2022 WHO grade 4) exhibited the highest age-adjusted incidence rates and is considered the most aggressive variant, characterized by an extremely aggressive biological behavior resulting in a poor outcome [2]. Despite the enormous progress in biotechnology and medicine field, life expectancy of glioblastoma (GB) patients has improved only slightly over the last 30 years. Recently, supramarginal resection or, where possible, excision of the hyperintense area in FLAIR sequences in MRI (so-called Flair-ectomy) has been shown to be associ- ated with improvement in both overall survival (OS) and progression free survivor (PFS), although not always execut- able, especially in lesions involving eloquent brain areas [5, 6]. Other therapeutic strategies tested included anti-angio- genic therapy and immunotherapy, though they did not show significant improvement in OS [7]. Some authors also advo- cated the importance of palliative care to increase the quality of life in patients affected from this tremendous tumor [8, 9]. As a matter of fact, if untreated, GB’s median sur- vival means is 3 months [3]. Interestingly, the standard f In the era of genomic and molecular genetics, in which it is possible to investigate all the potential metabolic land- scape of the disease, new treatment strategy is going to rely more on biochemical and immunological treatments [10–12]. Various new treatments have been proposed over the years, regarding use of CAR T cells [13], molecular agents enhancing the effect of radiotherapy (RT) [14], up to the application of high and low intensity focus ultrasound [15]. (0121 3456789) 3 220 Journal of Neuro-Oncology (2023) 163:219–238 In this context, sonodynamic therapy (SDT) seems to become a promising treatment, offering the possibility of non-invasively eradicate solid tumor in a site-directed manner, employing compounds that become cytotoxic after being exposed to low intensity ultrasound [16]. The importance of this kind of treatment could gain an added value considering the possibility of deep lesion-targeting thanks to the significant depth that low-intensity ultra- sound penetrates tissue, not damaging surround brain parenchyma and the chance of aiming directly to cancer stem cells (found to be vital cells for proliferation, dif- ferentiation, and treatment resistance of the GB) [17–20] future clinical trials, exploring the limitations and poten- tial of such treatment. Fig. 1 PRISMA Flow Diagram Search of the literature Preferred Reporting Items for Systematic reviews and Meta-analyses guidelines (PRISMA) were followed to conduct and report this systematic literature review [22] (Fig. 1). Remarkably, the possibility of impairing the brain blood barrier (BBB) is another weapon in neurosurgical arma- mentarium, making easier the access of chemical agents [20, 21]. We performed a broad systematic literature search in different online scientific libraries (Pubmed/MEDLINE, Cochrane Library and ClinicalTrial.Gov) for all studies investigating the efficacy and feasibility of SDT in GB treatment. The protocol of this review has been prospec- tively registered in Open Science Framework and it is pub- licly available online at https://doi.org/10.17605/OSF.IO/ FW4QS. Our review aims to provide a current view of the use of focused ultrasound, and particularly SDT, in the man- agement of GB, starting with their mechanism of action in vitro and in animal models and ending with current or Fig. 1 PRISMA Flow Diagram Total Records identified from: PubMed Database ® (n = 373) Records removed before screening: Duplicate records removed (n = 128) Records marked as ineligible by automation tools (n = 0) Records removed for other reasons (n = 0) Records screened (n = 245) Records excluded (n = 157) For title (n = 104) For Abstract (n = 53) Reports sought for retrieval (n = 88) Reports not retrieved (n = 0) Reports assessed for eligibility (n = 88) Reports excluded (n = 65) Focus on BBB disruption (n = 39) Systematic Review or Meta- analysis (n = 16) Other reasons (n = 10) Studies included in review (n = 23) Reports of included studies (n = 23) Identification Screening Included Identification of studies via databases and registers Fig. Results The studies included in our paper were both studies in vitro and in vivo using animal models, and ongoing clinical trials. The proof-of-concept of our research was trying to under- stand the effect of SDT on Glioma/GB cell line and then the feasibility, efficacy, and safety of its application firstly in animal model and then in the clinical practice, evaluating the ongoing clinical trials. Data extraction According to the criteria above, all articles were identi- fied by two reviewers (K.G. and U.E.B.). In case of a dis- crepancy, a third author (L.B.) arbitrated until a consensus among the authors was reached. To avoid the potential omission of relevant studies we also manually screened reference lists of articles included and previous systematic reviews and meta-analyses regard- ing the topic. Duplicate articles were eliminated using Microsoft Excel 16.37 (Redmond, WA, USA). The extracted data included the following: publication’s year, author, study design, patients’ number, patients’ mean age and gender, type of cells or animals, aim of the study and results of the study. Search of the literature 1 PRISMA Flow Diagram Identification of studies via databases and registers Identification of studies via databases and registers Records removed before screening: Duplicate records removed (n = 128) Records marked as ineligible by automation tools (n = 0) Records removed for other reasons (n = 0) Identification Total Records identified from: PubMed Database ® (n = 373) Reports sought for retrieval (n = 88) Reports not retrieved (n = 0) Reports excluded (n = 65) Focus on BBB disruption (n = 39) Systematic Review or Meta- analysis (n = 16) Other reasons (n = 10) Reports assessed for eligibility (n = 88) Studies included in review (n = 23) Reports of included studies (n = 23) 1 3 221 Journal of Neuro-Oncology (2023) 163:219–238 We searched for studies published up to the 15th of September 2022 without backward limits, using the fol- lowing MeSH terms “Glioblastoma” AND “Focused Ultra- sound”, “Glioblastoma” AND “FUS”, “Glioblastoma” AND “MRgFUS”, “Glioblastoma” AND “Sonodynamic therapy“, “Glioblastoma” AND “High intensity Focused Ultrasound”, “Glioblastoma” AND “HIFU”, “Glioblas- toma” AND “Sonodynamic” AND “therapy”, “Gliomas” AND “Sonodynamic therapy“, “Glioblastoma” AND “focused ultrasound” AND “sonosensitizer”, “Glioma” AND “focused ultrasound” AND “sonosensitizer”. • Editorials, books, systematic reviews, and meta-analysis. • Patients age < 18. • Editorials, books, systematic reviews, and meta-analysis. • Patients age < 18. • Patients treated with focused ultrasound used to perform a disruption in brain-blood-barrier (BBB). • Studies evaluating focused ultrasound therapy but not focusing on SDT. • Studies evaluating focused ultrasound therapy but not focusing on SDT. Data selection Our initial research carried out through Pubmed identified a total of 373 articles. We excluded 128 duplicated articles, then we performed a further screening based upon title and abstract reading, eliminating 157 articles. The research strategy initially relied on title and abstract analysis. The article’s full text was retrieved for further investigation if the title and abstract met the inclusion crite- ria. The data collection process was conducted without using any automated tools. The research was conducted by two different authors separately (U.E.B and K.G) and eventu- ally refined by a third author (L.B). No ethical approval was required for this study. Finally, after a full text reading and a detailed examina- tion, 65 articles were excluded, because either they were focusing only on the effect of focused ultrasound on BBB disruption (39 articles) or they were reviewing previous sci- entific works (16 articles) or lastly because they were not inherent with the purpose of this review, including finally 23 studies in our systematic review, according to PRISMA flow diagram inclusion criteria. l The characteristics of included articles are the following ones: publication’s year, author, study design, type of cells and animal model (respectively for the ‘in vitro’ and for the ‘in vivo’ studies), aim of the study and results of the study. Eligibility criteria The articles were selected according to the following inclu- sion criteria: The articles were eventually divided in three tables, including studies ‘in vitro’ and studies ‘in vivo’ on animal models (Tables 1, 2). • Full article in English. Using “Sonodynamic Therapy“ AND “Glioblastoma” and “Focused Ultrasound” AND “Glioblastoma” as MeSH terms, another research on ClinicalTrial.Gov was performed, identifying a total of 11 trials. After exclusion criteria were applied, only four trials were included in the review. • Studies in a preclinical phase (‘in vitro’ and ‘in vivo’ study). • Case report, case series, retrospective study, prospective study and clinical trials. • Patients age ≥ 18.f • Patients age ≥ 18.f The characteristics of included trials are the following ones: title of the trial, identifier, status, interventions charac- teristics, aim of the study and locations of the trial (Table 3). • Patients affected by glioblastoma treated with SDT. Exclusion criteria: Exclusion criteria: • Articles not in English. 1 3 Journal of Neuro-Oncology (2023) 163:219–238 222 1 3 Table 1 Summary of ‘in vitro’ studies included in the review References Study design Pub- blication year Type of cells Aim Parameters Results Outcome Hayashi et al. [23] In vitro 2009 U105MG and U251MG glioma cells To investigate the effi- cacy of low-intensity ultrasound on malignant astrocytic tumor cells using Photofrin as a photosensitizer Intensity: 0.3W/cm2 Photofrin-SDT enhanced US cell killing in LRP/ α2-MR-expressing glioma cells higher cell killing was observed following SDT: 52.7 ± 17.5%,13.0 ± 4.6%, and 3.9 ± 0.9% for 5, 15 and 30 s, respectively in comparison to US alone treatment Frequency: 0.95 MHz Duration of 5, 15, or 30 s Xu et. al. [24] In vitro 2012 U251 glioma stem-like cells To compare the suscepti- bility of GSCs and U251 to SDT using Photofrin as a sonosensitizer Frequency: 1.0 MHz Cell viability and apoptosis assays showed that SDT damaged both GSCs and U251 glioma cells, but GSCs were significantly less susceptible in the SDT groups, GSCs had a significantly lower apoptosis rate (p. < 0.01) than the U251 glioma cells (63.43 ± 8.07% vs. 38.85 ± 10.24%) Intensity:0.5W/cm2 Timing: 1 min Hao et al. Eligibility criteria [25] In vitro 2014 C6 glioma cells To investigate the role of intracellular calcium overload in the in vitro apoptosis of C6 glioma cells mediated by low level ultrasound and hematoporphyrin mono- methyl ether (HMME) therapy Frequency: 0.5 MHz The combined use of low-level ultrasound and HMME improved the apoptotic rate of C6 glioma cells mediated by ultrasound alone; it was showed that Ca2 overload plays a role in the SDT- induced apoptosis The SDT treatment in calcium-supplemented medium showed signifi- cant increase in apoptosis (49.4 ± 2.6%) in com- parison to control group (apoptotic rate < 5%) Intensity: 1.0 W/cm2 Timing: 1 min Gonzales et al. [26] In vitro 2016 F9 glioma cells To evaluate the efficacy of FUS activation of the sonosensitizer AIPcS2a(Aluminum phthalocyanine disul- fonate) together with Bleomycin Frequency: 1 MHz FUS activation of the sonosensitizer AlPcS2a together with BLM significantly inhibits the ability of treated glioma cells to grow as three-dimensional tumor spheroids SDT at an intensity of 0.6 W/cm2 causes a reduc- tion in spheroid volume by about 30%. In contrast, FUS alone at the same intensity leads to a reduc- tion in spheroid volume of less then 10% Intensity: 0.2, 0.4, or 0.6 W/cm2 Timing: 3 min Chen et al. [27] In vitro 2017 C6 glioma cells To evaluate the effects of combination of applica- tion of temozolomide (TMZ) together with SDT Frequency:1 MHz The combination between TMZ together with SDT enhanced the expression of mitochondrial pathway apoptosis proteins, as well as suppressed MMP-2 to weaken the migration ability in TMZ-resistant C6 cell line and by decreasing NHE1 protein expression The survival rates are 96.8 ± 7.9%, 94.5 ± 6.6%, 61.1 ± 13.3%,30.1 ± 4.4%. 28.5 ± 2.9%,26.3 ± 4.1%, 25.4 ± 4.2%, 19.3 ± 3.8% after 1, 2, 4, 6, 8, 12, 18, 24 h insonication respec- tively. Cell viability in SDT group was < 60% vs. 100% in control group Intensity: 1 W/cm2 Timing: N/A 1 Table 1 Summary of ‘in vitro’ studies included in the review Journal of Neuro-Oncology (2023) 163:219–238 223 Table 1 (continued) References Study design Pub- blication year Type of cells Aim Parameters Results Outcome Sun et al. Eligibility criteria [28] In vitro 2018 U373 human glioma cells and NIH 3T3 murine fibroblastic cells To evaluate the killing effect of SDT in human glioma cells by using sinoporphyrin sodium used as sonosensitizer Frequency:1.0 MHz The combined use of SDT with DVDMS reduced human glioma cells viability and apoptosis; no significant changes were observed in the viability of normal cells The cell viability was measured as 87.27%, 90.81% and 37.76% in the 2 microM DVDMS alone, 0.45 W/cm2 ultrasound and SDTgroup, respec- tively Intensity: 0.45 W/cm2 Timing: 1 min Dai et al. [29] In vitro 2019 C6 glioma cells To investigate the apoptotic effect and mechanisms of SDT mediated by hematoporphyrin mono- methyl ether (HMME) on C6 glioma cells in vitro Frequency:1 MHz SDT and HMME combina- tion increased the mito- chondrial signal pathway apoptosis; moreover, it was proved an increased production of ROS and a reduction of MMP The survival rates are 97.4 ± 2.4%, 83.1 ± 5.2%, 63.8 ± 4.2%, 33.8 ± 3.7%, and 27.4 ± 3.1% after 0, 30, 60, 90, and 120 s insonation, respectively. Apoptotic rate was > 35% in SDT group vs < 5% in control group Intensity: 1 W/cm2 Timing: varied from 0 to 120 s (interval of 30 s) Sheehan et al. [30] In vitro 2020 C6 glioma cells and U87 human GB cells To study the effects SDT using 5-Aminolevulinic acid hydrochloride (5-ALA) and high frequency focused ultra- sound (FUS) on 2 GB cell lines Frequency: 1 MHz SDT resulted in appreci- able GB cell death as compared to 5-ALA or FUS alone reductions in cell viability for 5-ALA, FUS, and SDT groups of 5%, 16%, and 47%, respectively com- pared to control Intensity: 10 W/cm2 Timing: 3 min Shono et al. [31] In vitro 2021 Mouse glioma stem cells To assess the anti-tumor effects of SDT with a Cox2 inhibitor in mouse glioma stem cells Frequency: 1 MHz The combination therapy with SDT and celecoxib resulted in enhanced anti- tumor efficacy among GSCs and a mouse GSC- bearing glioma model Tumor volume reduction at 14 days after SDT was > 70% vs < 40% in control group. Kaplan– Meier survival rate and survival period prolonged by the combination therapy with SDT and celecoxib Intensity: 2 W/cm2 Timing: 2 min Shen et al. Eligibility criteria [32] In vitro 2021 U87MG human GB cells To investigate the anti- tumor effect of SDT combined DVDMS on human GB Frequency: 1 Hz The DVDMS-mediated SDT could induce great cytotoxicity in GB cells via the production of ROS The apoptosis and necro- sis ratio of SDT group: 29.17 ± 3.45%. Survival cell rate was < 40% in SDT group vs > 95% in control group Intensity: 0.32W/cm2 Timing: 3-min Journal of Neuro-Oncology (2023) 163:219–238 224 1 3 Table 2 Summary of in vivo studies included in the review References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Nonaka et al. [33] In vivo 2009 C6 glioma cells Wistar rats To determine the optimal focused ultrasound acoustic condi- tions using the photosensitizer Rose Bengal (disodium tetra- iodo tetrachloro fluorescein) for the ablation of experimental intracranial glioma in rats Frequency: 1 MHz The combination of SDT and Rose Bengal showed a selective antitumor effect against cerebral glioma while sparing normal brain tissues The areas of lesions in animals with and without Rose Bengal at an inten- sity of 110 W/cm2 for 5 min were 1.84 ± 0.18 and 4.47 ± 0.70 mm2, respectively (p < 0.05) Intensity: 25 W/ cm2 Timing: 5 min Ohmura et al. [34] In vivo 2011 C6 glioma cells Wistar rats To investigate the antitumour effect of SDT in experi- mental rat glioma by using focused ultrasound in combination with 5-ALA used as a sonosensitizer Frequency: 1.04 MHz SDT with 5-ALA and focused ultrasound achieved selec- tive antitumour effect against deep-seated experimental glioma The largest tumour areas found were 29.94 ± 10.39, 30.81 ± 9.65, 32.98 ± 7.21 and 18.32 ± 5.69 mm2 in the rats undergoing sham operation, ultrasound irradiation alone, 5-ALA alone and SDT, respec- tively Intensity:10,15, 20 or 25 W/cm2 Timing: 5 min Jeong et al. Eligibility criteria [39] In vitro and in vivo 2018 U87 glioma cells and U251 glioma stemlike cells Immunodeficient mice To investigate the antitumor activity of SDT combined with a sonosensitizer, 5-aminolevulinic acid (5-ALA), on malignant gliomas Frequency: 3-MHz 5-ALA-SDT inhibited cell growth, changed cell morphol- ogy, and induced apoptosis; in vivo 5-ALA-SDT with HIFU pro- longed survival of the tumor- bearing mice Authors demonstrated that the mice treated with 5-ALA-SDT by using HIFU survived much longer than either the untreated mice (control) or the mice treated with HIFU alone Intensity: 2 W/cm2 Timing: 3 min Pi et al. [40] In vitro and in vivo 2019 U87 MG-Red- FLuc human GB cells Male Balb/c nude mice To study the anti-tumor effect of sinopor- phyrin sodium (DVDMS)- mediated SDT on nude mice bear- ing intracranial U87 MG-Red- FLuc human GB Frequency: 0.996 MHz DVDMS-mediated STD leads to an anti-proliferation effect and cell apoptosis induc- tion the bioluminescence photon emission value of SDT w/DVDMS + FUSiBBBo group (1.59 ± 0.68 × 107 photons/s) was sig- nificantly lower than the control group (115.1 ± 97.6 × 107 photons/s) on the 23rd day after tumor implanta- tion Intensity: 1.7 W Timing: 1 min Table 2 (continued) References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Ju et al. [38] In vitro and in vivo 2016 SNB19 and U87MG glioma cells BALB/c nude mice To investigate whether hyper- thermotherapy (HT) could improve the effi- cacy of SDT in Xenograft tumor in nude mice model Frequency: 1 MHz The combination of SDT, HT and sonosensi- tizer produced a reduction of cell viability, increased apop- tosis through increased ROS production, loss in MMP, higher protein levels of Bax and cleaved caspase-3, 8, and 9 and sig- nificantly lower protein level of bcl-2 the tumor volume in HT plus SDT group is much smaller compared with SDT group (P < 0.05) Intensity: 2.0 W/ cm2 Timing: 10 min Suehiro et al. Eligibility criteria [35] In vivo 2012 C6 glioma cells Sprague Dawley Rats To evaluate the sonodynamically induced selective antitumor effects of 5-aminole- vulinic acid (5-ALA) on a C6 glioma that was implanted in a rat brain Frequency: 1.0 MHz SDT with 5-ALA and focused ultrasound irradiation was effective for the treatment of intracranial gliomas in rats, by a selective tumor destruc- tion and tumor growth inhibition through nonther- mal effects of weak ultrasound irradiation in a fractionated manner a 67% or 53% reduction in tumor volume occurred in the groups receiving 5-ALA-SDT ultrasound irradiation either 6 h or 3 h after the 5-ALA administration, respec- tively Intensity: 2.65 W/ cm2 Timing: 20 min 225 Journal of Neuro-Oncology (2023) 163:219–238 Table 2 (continued) References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Tserkoorvsk et al. [36] In vivo 2012 C6 glioma cells Rat To investigate the effect of SDT using Photolon as a sonosensi- tizer for the abla- tion of glioma C6 tumor model in rats Frequency: 1 MHz The combination between ultra- sounds and sono- sensitizer agent leads to strong damage of tumor tissue because of developing induced chemi- cal reactions and cavitation effect implementation in the tumor cell Photolon + ultrasound 0.7 W/cm2 + Photoirradiation 100 J/cm2 caused an area of necrosis of 100% Intensity: 0.7 W/ cm2 Timing: 10 min Song et al. [37] In vitro and in vivo 2014 C6 glioma cells Wistar rats To investigate the effect of hematoporphyrin monomethyl ether (HMME)- mediated SDT on C6 gliomas implanted in rat brains Frequency: 1 MHz SDT treatment could inhibit the expansion of intracranial gliomas related to mechani- cal injury and the induction of apoptosis, mediated by the increased levels of Cyto-C, microvessels dis- truction, inhib- ited angiogenesis and increased expression of HIF1 Tumor volume: 50 mm3 (pre-op); 15mm3 vs 100 mm3 (1-week post-op, SDT group vs control group, respectively); 55 mm3 vs 140 mm3 (2 weeks post-op, SDT group vs control group, respectively) Intensity: 0.5 W/ cm2 Survival time: > 50 days (SDT group) vs < 30 days (control group) Timing: 120 s Song et al. [37] In vitro and in vivo 2014 1 3 Journal of Neuro-Oncology (2023) 163:219–238 226 Journal of Neuro-Oncology (2 protein level of bcl-2 Suehiro et al. Eligibility criteria [39] In vitro and in vivo 2018 U87 glioma cells and U251 glioma stemlike cells Immunodeficient mice To investigate the antitumor activity of SDT combined with a sonosensitizer, 5-aminolevulinic acid (5-ALA), on malignant gliomas Frequency: 3-MHz 5-ALA-SDT inhibited cell growth, changed cell morphol- ogy, and induced apoptosis; in vivo 5-ALA-SDT with HIFU pro- longed survival of the tumor- bearing mice Authors demonstrated that the mice treated with 5-ALA-SDT by using HIFU survived much longer than either the untreated mice (control) or the mice treated with HIFU alone Intensity: 2 W/cm2 Timing: 3 min Pi et al. [40] In vitro and in vivo 2019 U87 MG-Red- FLuc human GB cells Male Balb/c nude mice To study the anti-tumor effect of sinopor- phyrin sodium (DVDMS)- mediated SDT on nude mice bear- ing intracranial U87 MG-Red- FLuc human GB Frequency: 0.996 MHz DVDMS-mediated STD leads to an anti-proliferation effect and cell apoptosis induc- tion the bioluminescence photon emission value of SDT w/DVDMS + FUSiBBBo group (1.59 ± 0.68 × 107 photons/s) was sig- nificantly lower than the control group (115.1 ± 97.6 × 107 photons/s) on the 23rd day after tumor implanta- tion Intensity: 1.7 W Timing: 1 min 1 3 ( ) References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Ju et al. [38] In vitro and in vivo 2016 SNB19 and U87MG glioma cells BALB/c nude mice To investigate whether hyper- thermotherapy (HT) could improve the effi- cacy of SDT in Xenograft tumor in nude mice model Frequency: 1 MHz The combination of SDT, HT and sonosensi- tizer produced a reduction of cell viability, increased apop- tosis through increased ROS production, loss in MMP, higher protein levels of the tumor volume in HT plus SDT group is much smaller compared with SDT group (P < 0.05) Intensity: 2.0 W/ cm2 Timing: 10 min 3 227 Journal of Neuro-Oncology (2023) 163:219–238 Table 2 (continued) References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Sun et al. Eligibility criteria [41] In vitro and in vivo 2019 C6 glioma cells Balb/c mice To evaluate the efficacy of nanosensitizer- DVDMS (sinoporphyrin sodium) loaded iRGD modi- fied liposomes (iRGD-Lipo- DVDMS) as a potential drug delivery system, combined with low-intensity focused ultra- sound (FUS) for glioma SDT Frequency: 1 MHz The combination of DVDMS- SDT produced an anti-tumoral effect through increased cell apoptosis the median survival time of the glioma bearing mice treated with iRGD-Lipo- DVDMS-SDT (40 days) was obviously longer than that of those treated with only saline (15 days), free DVDMS (19 days), or LipoDVDMS-SDT (24 days) Intensity: 0.4, 0.6 and 0.8 W/cm2 Timing:1 min Yoshida et al. [42] In vitro and in vivo 2019 F98 glioma cells Female Fisher rats To investigate the efficacy of 220- kHz TcMRgFUS combined with 5-aminolevulinic acid (5-ALA) on malignant glioma in vitro and in vivo Frequency: 220 kHz FUS/5-ALA com- bination reduced cell viability by inducing apopto- sis and sup- pressed tumor proliferation and invasion as well as angiogenesis in vivo, while causing minimal damage to nor- mal brain tissue Viability was decreased by the combination of FUS and 5-ALA at a total energy 3000 J relative to irradiated cells in the other groups (p < 0.01), reflecting a total energy dependence for the cyto- toxic effects of SDT Intensity: 10-20W Timing: 120–240 s An et al. [43] In vitro and in vivo 2020 U-118 MG and U-87 MG human glioma cells BALB/c nude mice To explore the antitumor effects of sinopor- phyrin sodium (DVDMS)-medi- ated photody- namic therapy (PDT) and SDT in glioma Fre- quency:1.0 MHz DVDMS-mediated PDT and SDT inhibit the prolif- eration of glioma cells and induce apoptosis, inhibit tumor develop- ment in U-118 MG xenograft models, sup- pressed PCNA and Bcl-xL levels the fluorescence intensity of DVDMS was lower in the PDT and SDT groups compared with the DVDMS group, while tumor cell proliferation and weight were lower in the PDT and SDT groups than in the control group Intensity: 500 mW/ cm2 Timing:1 min Yoshida et al. [42] In vitro and in vivo 2019 1 Journal of Neuro-Oncology (2023) 163:219–238 228 Study characteristics ( ) References Type of study Publication year Type of cells Animal model Aim Parameters Results Outcome Prada et al. [44] In vivo 2020 C6 glioma cells Sprague–Dawley rats To demonstrate the efficacy of SDT with fluorescein (FL) and low- intensity focused ultrasound in inhibiting the growth of ectopic gliomas implanted in the rat's subcutane- ous tissue Frequency: 350 kHz SDT signifi- cantly inhibited outgrowth of ectopic C6 gliomas across FUS exposure conditions SDT significantly restricted tumor outgrowth across all three FUS exposure conditions compared to fuorescein alone (p = 0.001, p < 0.0001, and p < 0.0001for FUS1x, 1.5 × , and 2 × + Fluo, respectively) Intensity: 2–3 W/ cm2, or 3–4.5 W/ cm2, or 4–6 W/ cm2 Timing: 10 ms Raspagliesi et al. [45] In vivo 2021 C6 glioma cells Pigs To demonstrate the safety of low- intensity ultra- sonic irradiation of healthy brain tissue after the somministration of a sonosensitizer Frequency: Low frequency SDT and the use of Na-FL or 5-ALA as sonosesitizer are safe towards healthy brain tissue Upon histopathological examination, the targeted regions did not exhibit any notable pathological abnormalities. Both neu- rons and glial cells were found to be intact, with no significant cellular alterations observed Intensity: 2–3 W/ cm2 Timing: 20 min In vitro' and in vivo' studies Tumor cell lines used in the listed studies were both murines and humans: rat C6 glioma cells were the most used ones, followed by human U87 GB cells and other cell lines such as human glioma cells U373, U105MG and U251MG. ‘In vivo’ studies used as animal models mainly murines, both mice and rats; only one study employed a porcine model.f Both ‘in vitro’ and ‘in vivo’ studies used different types of sonosensitizers, such as 5-Aminolevulinic acid hydrochlo- ride (5-ALA), sinoporphyrin sodium (DVDMS), hemato- porphyrin monomethyl ether (HMME), temozolomide (TMZ), photofrin, fluorescein (FL) and disodium tetraiodo tetrachloro fluorescein (Rose Bengal), as a way of increas- ing the tumoral cells’ vulnerability to focused ultrasounds’ exposition.f ‘In vitro’ studies focused their attention on the effect that ultrasound, with or without the use of a sonosensitizer, pro- voked on tumoral cell lines, in term of apoptotic rate and intracellular level of reactive oxygen species (ROS) post- exposition; moreover, the minimal intensity of ultrasound in order to produce an apoptotic effect on tumoral cell was also investigated [23]. Some studies also tried to quantify the anti-tumoral effect of ultrasound with or without the use of a sonosensitizer [26].fi of a sonosensitizer Moreover, Gonzales et. al. proved the increased efficacy of STD in combination with bleomycin, in its inhibition of tumoral growth. ‘In vivo’ studies used an animal model to verify the fea- sibility of this technique: more than investigating just the anti-tumoral effect of STD, these studies prove the safety of SDT towards healthy brain tissue [33, 45]. Other information obtained from ‘in vivo’ studies regard the efficacy of focused ultrasound used in combination of a sonosensitizer in inducing tumor growth inhibition and the underlying physiopathological mechanism, described thanks to post-autoptic histology. STD therapy was able to induce an increased apoptotic rate, through an increased ROS pro- duction, reduced production anti-apoptotic/pro-angiogenic factors and microvessel destruction [37]. Many of the ‘in vivo’ studies were coupled to in vitro experiments where the same method was tested, report- ing analogies and differences in both results; some studies, instead, were performed directly ‘in vivo’, on animal mod- els. The studies reviewed used similar sonication parameters regarding intensity of sonication performed (range from 0.2 to 25 W/cm2). The frequency used in the studies ranged from 0.5 to 3 MHz. The maximum value of the duration was 20 min. All the studies included, especially ‘in vivo’ stud- ies, have demonstrated that SDT are effective in reducing 3 Journal of Neuro-Oncology (2023) 163:219–238 229 Table 3 Ongoing clinical trials on SDT in Glioblastoma Title Identifier Status Interventions Aim Locations Sonodyamic therapy with exablate system in glioblastoma patients (sonic ALA) NCT04845919 not yet recruiting 5-Aminolevulinic acid [5-ALA] To evaluate the safety and feasibility of SDT with 5-aminolevulinic acid in patients with newly diagnosed cerebral GB using the ExAblate Model 4000 Type-2 “Neuro- System” Fondazione I.R.C.C.S. Istituto Neuro- logico Carlo Besta, Milano Study of Sonodynamic Therapy Using SONALA-001 and Exablate 4000 Type 2 in Recurrent GB NCT05370508 Recruiting SONALA-001 (ALA) and exablate device To evaluate the safety, dose-limiting toxicities, recommended Phase 2 schedule, and preliminary efficacy of SDT using SONALA-001 and Exablate Type-2 device in subjects with recurrent or progressive GB Ivy Brain Tumor Center, Ari- zona and MD Anderson Cancer Center,Houston Study to Evaluate 5-ALA Combined with CV01 Delivery of Ultrasound in Recurrent High-Grade Glioma (HGG) NCT05362409 Recruiting 5-Aminolevulinic acid [5-ALA] To evaluate the Safety and Toler- ability of 5-aminolevulinic Acid (5-ALA) Combined with CV01 Delivery of Ultrasound for SDT in Patients With recurrent HGG Northwell. of a sonosensitizer United States, New York Study of Sonodynamic Therapy in participants with recurrent high- grade glioma (HGG) NCT04559685 Recruiting SONALA-001 (ALA) To evaluate the ascending energy doses of SDT utilizing the Mag- netic Resonance focused Ultra- sound (MRgFUS) combined with intravenous 5-ALA and efficacy in patients with recurrent HGG St. Joseph's Hospital and Medical Center 1 3 Journal of Neuro-Oncology (2023) 163:219–238 230 therapy, in combination with chemotherapy or radiation therapy. The main points of values of FUS are the non- invasiveness, incision-free, controllability via real-time MR guidance and the capacity to activate the immune sys- tem [54]. The first non-invasive thermal ablation of a brain tumor in human was realized by Coluccia et al. [55] in their ongoing clinical phase I study in 2014, when they firstly employed Magnetic resonance-guided focused ultrasound surgery (MRgFUS) for safe thermal ablation of a centrally located recurrent GB. This is possible thanks to recent advances in magnetic resonance imaging, which allow safe and precise thermal ablation of neoplastic tissue. Moreover, the opportunity to create an MRI-derived temperature map- ping of the targeted tissue allow a non-invasive monitoring of the ablating procedure. In more recent years, knowledge about the different mechanisms of action of ultrasound at various intensities and frequencies, used alone or in combi- nation with other substances, has been expanded, exploring new potentials, and developing new therapeutic strategies, including precisely SDT [56–61]. tumor volume, because of its high selectivity, low toxicity, and deep penetration, focusing on both the ability to reduce tumor growth and placing emphasis on the survival of tumor cells after the treatment (Tables 1, 2). Sonodynamic therapy Finally, we report a phase 0 single-center open label study whose intention is to appraise the ascending energy doses of SDT utilizing the MRgFUS combined with intravenous 5-ALA and its efficacy in patients with recurrent HGG. SDT has been developed as a promising tool in brain tumor treatment. SDT takes its cue from photodynamic therapy (PDT), in which a light-activated photosensitizer can cause the generation of ROS, which in turn would mediate a cyto- toxic effect on neoplastic cells. However, the main limitation of PDT is the range of action, which is limited to superficial lesions due to the poor penetration of laser light into brain tissue [42, 62]. This obstacle is overcome using low-intensity ultrasound, which has a greater penetrative capacity [63]. fi Studies characteristics and aim were summarized in Table 3. Clinical trials We identified four clinical trials about SDT in GB treatment. Between them, it is listed a non-randomized, single-arm study whose purpose is to evaluate the safety and feasibility of SDT with 5-aminolevulinic acid in patients with newly diagnosed cerebral GBs using the ExAblate Model 4000 Type-2 Neuro System. Another clinical trial, non-randomized, tried to assess the safety, dose limiting toxicities, and preliminary efficacy of SDT using SONALA-001 and Exablate Type-2 device in subjects with recurrent or progressive GB. Additionally, a phase 1 multi-center trial started to under- stand the safety and tolerability of 5-aminolevulinic acid (5-ALA) combined with CV01 delivery of ultrasound for SDT in patients with recurrent high-grade glioma. 1 3 Discussion Conventional therapeutic options in the treatment of solid brain tumors, and GBs in particular, are based on the assumption that these cancerous lesions have relatively homogeneous spatiotemporal characteristics. However, recent advances in the molecular, genetic, and epigenetic fields have shown how this does not reflect the facts at all, underscoring the inherent limitations of radiation and chem- otherapy [46–48]. Moreover, the notion that GBs do not represent a focal pathological entity, but rather a pathology spread throughout the entire brain, makes clear the inherent limitations of surgery, although it still represents the thera- peutic mainstay toward these tumors [49, 50]. As just mentioned, SDT involves the application of focused ultrasound with a substance that sensitizes cells to the destroying effects of sound, called sonosensitizer. It includes both ultrasonication, via non-invasive low-intensity ultrasound penetrating soft tissues and focus on a specific site, and sonosensitizers, which embrace non-toxic chemi- cal agents such as 5-ALA, ATX-70, Hypocrellin, Rose Ben- gal and many others [64–66]; some of these compounds are commonly used in glioma surgery to intraoperatively visualize the tumor and can be employed to induce cyto- toxic effects to neoplastic cells when subjected to a specific acoustic field [31, 39, 40]. The advantage of this technique is to minimize adverse events and maximizing on-target responses. Furthermore, the use of chemical agents that are non-toxic in the absence of a specific stimulus distinguishes the definition of SDT from the broader meaning of FUS employed to enhance the effects of an already toxic com- pound [67, 68]. Hence the need to develop new therapeutic strategies capable of eradicating the underlying pathology, possibly in the least invasive way, and increasing OS and PFS while safeguarding patients' quality of life and neurological status [51, 52]. In this context, the use of ultrasound for therapeutic pur- poses (the so-called Theranostics) appears to offer interest- ing potential and promising results and uses [53]. Sheehan et colleagues [69] employed the SDT on two cellular lines, rat C6 and human U87 GB cells, and found that two innocuous agents, which are FUS and 5-ALA, As a matter of fact, focused ultrasound can be employed either to destroy cancerous cells by heating or as an adjuvant 1 3 Journal of Neuro-Oncology (2023) 163:219–238 231 can lead to cell death by the transformation of 5-ALA to PPIX in malignant glioma cells, where it generates reactive oxygen species responsible of cellular apoptosis. Mechanism of action of sonodynamic therapy The effects of thermal ablation on tumoral cells are still not completely clear. Is has been demonstrated that hyperther- mia (HT) can enhance the 5-ALA-SDT induced cell apop- tosis partly by activating caspases and by modulating Bcl-2 family members. Moreover, HT is responsible of increasing ROS production and reducing metalloproteases (MMPs) induced by 5-ALA-SDT in human glioma cells [38, 80] (Figs. 2, 3). In 2019, Abdolhosseinzadeh et al. [70] investigated the effects of focused acoustic waves in a focal area through some accurate simulations. Results obtained in 2D, and 3D models showed that ultrasound waves could be used in the form of pulse waves with different time periods to provoke a focused thermal lesion on neoplastic tissue [71, 72]. Never- theless, ‘in vivo’ it is necessary to overcome the blood brain barrier (BBB), which represents a real obstacle to sonosen- sitizers. To this aim, low-intensity focused ultrasound can be combined to microbubbles, which proved to increase the permeability of the BBB, allowing the treatment of intrac- ranial GB in mice. These results can suggest the use of SDT with sonosensitizers in human GB [73–75]. HT can also regulate some molecular aspects of the immune response, such as Fas gene and its ligand FasL, and act as an immunomodulator in cancer therapy [81, 82]. In more details, it seems that the increasing in local temperature may act as a natural trigger or danger signal to the immune system. Hyperthermia can therefore enhance the expression of FAS-ligand mRNA, which has a role in functional maturation of dendritic cells together with secre- tion of proinflammatory cytokines, which in turn activate T lymphocytes and induce a polarization toward a Th1 phenotype. Moreover, high temperature may promote the action of a particular set of protein, the HSPs, which may act in protecting cells from dangerous stress by regulating cell homeostasis [83] and may also affect the stability of cellular membranes by inducing structural changing that intervene in signaling events and cell migration in immune response [84, 85]. Discussion ‘In vivo’ studies have proved that focused ultrasound in combination with the systemic administration of 5-ALA is effective in treating intracranial gliomas in rats, not demonstrated by the complete tumor resection but by the reduction in tumor size from the initial tumor volume [34, 35, 37, 39]. In this regard, Nonaka et al. [33] pinpointed the optimal focused ultrasound acoustic energy and duration for the ablation of brain tumor in rats, without damaging normal brain tissue; in their experience, this selective anti-tumor effect was pro- duced by weaker focused ultrasound intensity at 25 W/cm2 at 1 MHz for 5 min. can lead to cell death by the transformation of 5-ALA to PPIX in malignant glioma cells, where it generates reactive oxygen species responsible of cellular apoptosis. ‘In vivo’ studies have proved that focused ultrasound in combination with the systemic administration of 5-ALA is effective in treating intracranial gliomas in rats, not demonstrated by the complete tumor resection but by the reduction in tumor size from the initial tumor volume [34, 35, 37, 39]. In this regard, Nonaka et al. [33] pinpointed the optimal focused ultrasound acoustic energy and duration for the ablation of brain tumor in rats, without damaging normal brain tissue; in their experience, this selective anti-tumor effect was pro- duced by weaker focused ultrasound intensity at 25 W/cm2 at 1 MHz for 5 min. presence of a lesion where the latter can reach a significant concentration. This concept led to the non-invasive effect on SDT on normal brain tissue, since, even if the sonosensitizer has been collected in healthy tissue, it would be inconse- quential [45]. Thus, it seems clear that the choice of sonosensitizer is also crucial. Ideally, the perfect sonosensitizer should exhibit high affinity for tumor cells and slow clearance from the neoplasm, while sparing healthy brain parenchyma [77–79]. Ultrasound parameters affecting SDT results Interestingly, SDT can also be used in combination with other agents commonly used to treat GB to enhance their action, such as the temozolomide (TMZ). Resistance of high-grade glioma cells to TMZ is related to high level expression of NHE-1 protein, which enhance cells invasion to normal brain tissue. In their article, Chen et al. [27] dem- onstrated that SDT can suppress NHE-1 expression, thus allowing the cytotoxic effect of TMZ in vitro. Although the exact mechanism of action of SDT is not yet fully understood, it is assumed that the biological effects of this technique are strongly correlated with the phenomenon of acoustic cavitation (stable vs. inertial cavitation) derived from the interaction between ultrasound and the propagation medium, ultimately resulting in apoptosis of the affected cells. In addition to the mechanical effect of ultrasound, the action of SDT is also based on the sonochemical effect related to the formation of various species of free radicals and the different decomposition kinetics of sonosensitizers [63, 95, 96]. These various mechanisms of action in turn are closely related to the ultrasound parameters used and to other factors associated to the experimental setting (see Table 4). f Another target to take advantage of to enhance the anti- tumor efficacy of TMZ is the p-glycoprotein, referred to as multidrug resistance receptor (MDR1), a transmembrane protein which act as an efflux pump and confer multidrug resistance in brain tumor. Consequently, high expression of MDR1 is present in resistant GB and the downregulation of MDR1 via Akt/NF-kB pathway can improve the antitumor effect of temozolomide in GB cells. Shono et al. [31] dem- onstrated that the elevation of cellular PpIX using celecoxib is related to a down regulation of Akt/NF-kB/MDR1 path- way, thus enhancing the anti-tumor efficacy of SDT. Some authors advocated that the SDT mediated by hematopor- phyrin monomethyl ether (HMME) can induce apoptosis on C6 glioma cells in vitro and suggest that the mitochon- drial signal pathway may play a pivotal role, because of the observed production of ROS, loss of MMP and Bcl-2 and protein expression in caspace-9, caspase-3 and Bax [25, 29]. For instance, it has been demonstrated that most sonosen- sitizers respond to US frequency ranging from 0.2 to 3 MHz [42] and that a decrease in frequency is correlated to an increase in ultrasound toxicity [25]. Sonosensitizers As previously mentioned, the sonosensitizers used in SDT are harmless molecules that when subjected to an acoustic field mediate a cytotoxic effect. Many of these molecules are the same as those used in photodynamic therapy and are agents based on porphyrin or related molecules (protopor- phyrin IX, hematoporphyrin, etc.). In fact, there is evidence that such molecules, when exposed to the action of ultra- sound, result in the production of reactive oxygen species (ROS). In their ‘in vitro’ study, Shen et al. [32] employed as a sonosensitizer the sinoporphyrin sodium, purified from photofrin II, which showed great antitumor effect on human GB cell lines; particularly, this sonosensitizer can easily enter in cancer cells and accumulate into the mitochondria, where it gives raise to cytotoxity through the production of ROS. The link between ultrasound exposure, presence of sono- sensitizer and generation of ROS appear clear, and there is a consensus regarding their involvement in mediating the cytotoxic effect on cancer cells [86–88]. Nonetheless, other mechanisms have been proposed to elucidate the SDT- mediated cytotoxic effect. These include sonoluminescence, namely the emission of light from cavitation bubbles, which would appear to play a role both in the activation of certain sonosensitizers and in mediating antitumor effects [89, 90], and sonomechanical mechanisms that would mediate dam- age by inducing changes at the level of cell membranes, such as a reduction in membrane fluidity and an increase in lipid peroxidation [91, 92]. Noteworthy are the various cytotoxic actions and implicated mechanisms that characterize the dif- ferent sonosensitizers, although further studies on this are needed [93, 94]. However, although these agents are preferentially picked up by the tumor, they exhibit marked hydrophobicity, and their distribution appears to be ubiquitous [76]. Despite this apparently drawback, it has been postulated by Raspa- gliesi and colleagues that three contemporary events must occur to determine a cytotoxic effect: the administration of ultrasound, the administration of a sonosensitizer, and the 1 3 3 232 Journal of Neuro-Oncology (2023) 163:219–238 Ultrasound parameters affecting SDT results However recent works have pointed out apoptotic cell ratio was primarily affected by sonosensitizer concentration and then by other variables such as US frequency, irradiation time and intensity [97, 98]. US intensity usually ranges from 0.5 to 10W/cm2 and can be applied in a continuous or pulsatile mode [96]. Regarding this parameter, many studies have noted an intensity-depend- ent reduction in cell viability of various cancer types [99, ROS ROS ROS Fig. 2 Sonodynamic therapy could be effective in glioma cells-death though transformation of 5-ALA (green dots) to PPIX (red dots). The pro- cess results in the production of reactive oxygen species (ROS), leading to cells death Fig. 2 Sonodynamic therapy could be effective in glioma cells-death though transformation of 5-ALA (green dots) to PPIX (red dots). The pro- cess results in the production of reactive oxygen species (ROS), leading to cells death 3 233 Journal of Neuro-Oncology (2023) 163:219–238 Fig. 3 Representation of 3 mechanisms of sonodynamic therapy: A 5 ALA SDT could reduce level of Bcl2, thus activating apoptosis via caspase 9 pathway. B 5 ALA SDT increase levels of ROS, thus inducing cell death. C heating itself could increase level of Heat shock protein 70, thus inducing immune response through structural changing in cell membrane BCL- 2 CELL DEATH Caspase 9 ROS ROS ROS ROS 5 ALA - SDT 5 ALA - SDT Heang Hsp70 Immune response ultrasound; 5-ALA will be then re-administered every 4 weeks prior to CV01. The primary outcome is to evalu- ate the incidence of adverse events and to determine the Maximum Tolerable Duration in the first 12 months. Sec- ondary outcome is represented by the assessment of Overall 100]. Nejad et al. [101] have shown in a model of human oral squamous cell line HSC-2 how irradiating cells with 3.5 MHz US at 20, 32, 55, and 73 W/cm2 was associated to a cell survival rate of 97, 81, 62 and 40%, respectively. Irradiation time and duty cycle also influenced SDT results, with greater citotoxity at greater irradiation time and duty cycle [102]. Besides US parameters, many other fac- tors may influence SDT response ‘in vitro’ studies. Irradia- tion uniformity and intensity distribution, for example, are related to the distance between cells and US apparatus, as well as the characteristics of the coupling media and culture medium also seem to profoundly impact the outcomes of SDT. Ultrasound parameters affecting SDT results Other two crucial factors that affect the results of SDT are the type of irradiated sample and the sonosensitizer con- centration, the latter closely related to the apoptotic effect, as shown by Zhang et al. [103–105]. Table 4 ‘In vitro’ factors and variables potentially influencing SDT outcomes Table 4 ‘In vitro’ factors and variables potentially influencing SDT outcomes Factors Variables Ultrasound Parameters • Frequency • Intensity • Irradiation time and duty cycle Spatial Configuration • Cell-to-transducer distance • Ultrasound beam/culture vessel ratio Coupling Media • Volume • Composition • Viscosity • T° • Acoustic Propagation Culture Vessels • Geometry • Absorbers • Material Type Culture Medium • Composition • Volume • Acoustic Properties Irradiation sample type • Adhering cells • Suspension cells • Cell motion Fig. 3 Representation of 3 mechanisms of sonodynamic therapy: A 5 ALA SDT could reduce level of Bcl2, thus activating apoptosis via caspase 9 pathway. B 5 ALA SDT increase levels of ROS, thus inducing cell death. C heating itself could increase level of Heat shock protein 70, thus inducing immune response through structural changing in cell membrane Ongoing clinical trials Evidence regarding the potential application of SDT in high- grade gliomas are mostly taken from pre-clinical ‘in vitro’ and ‘in vivo’ studies. Currently, there are four ongoing trials concerning the role of SDT in high grade gliomas registered on “Clinicaltrial.gov”, of which three are recruiting.i The aim of the first trial is to evaluate the safety and tol- erability of 5-ALA combined with CV01 delivery of ultra- sound in patients affected by recurrent high-grade gliomas. (ClinicalTrials.gov Identifier: NCT05362409). This ongoing phase 1 trial is recruiting 33 patients, to which 5-ALA will be administered as sonosensitizer prior to CVo1-delivered ultrasound, which will deliver non-ablative, low-intensity 1 3 Journal of Neuro-Oncology (2023) 163:219–238 234 valuable option in patients with GB, due to the opportunity to induce toxicity only in a precise localization while mini- mizing harm in normal areas. Indeed, thanks to the devel- opment of increasingly sophisticated and accurate software is possible to target tumor volume precisely. In addition, nanotechnology-based drug delivery systems have been developed to enhance the selective accumulation of the sonosensitizer in tumor cells. Actually, SDTseems to be more effective in treating GB than low-grade glioma (LGG). This is because GB cells are more susceptible to the effects of SDT due to their higher rate of metabolism and greater degree of angiogenesis compared to LGG cells [38, 106, 107]. However, more research is needed to confirm these findings and determine the optimal parameters for SDT in the treatment of different types of brain tumors. Further- more, the effectiveness of SDT may also depend on other factors such as tumor size, location, genetic characteristics, and vascular pattern. For instance, brain tumors located near the skull base may be more amenable to SDT. The size of the tumor can also affect the effectiveness of the procedure. Larger tumors may be more difficult to treat with SDT, as the ultrasound waves may not be able to penetrate deep enough into the tumor to effectively kill the cancer cells. Response Rate, Duration of Response, OS and PFS in the first 12 months. i The second recruiting trial (ClinicalTrials.gov Identifier: NCT05370508) aims to evaluate the safety and preliminary efficacy of SDT by using SONALA-001 as sonosensitizer and Exablate 4000 Type-2 MR-guided focused ultrasound as device in people affected by recurrent or progressive GB. Ongoing clinical trials The primary outcomes are represented by the evaluation of the safety of SDT in the first 12 months, the maximum tolerable duration in the first 29 days, the determination of recommended phase 2 schedule and the assessment of pro- gression free survival in the first 6 months. i The third prospective, non-randomized, single-arm, not yet recruiting study (ClinicalTrials.gov Identifier: NCT04845919) aims to evaluate the safety and feasibility of SDT by using 5-ALA and Exablate 4000 Type-2 MRg- FUS in patients newly diagnosed with GB. Patients screened will undergo SDT treatment, will perform a strict neuro- radiological follow-up after the procedure (minimum 2 MRI) and will undergo tumor resection 14–21 days after SDT. The primary outcome is represented by the early identification of hemorrhage, oedema, or other damages in the first 10 days; secondary outcomes are represented by the evaluation of the rate of neurological deficits and the radiological response to treatment in the first 10 days after the procedure.i f Limitation on the clinical application rely on the fact that SDT represents a novel technique that needs to be fur- ther investigated. First, the role of sonosensitizers should be deepened: many sensitizers are employed in both PDT and SDT and residuals can accumulate in areas other than tumors, thus leading to hypersensitivity to light. Strategies to overcome this limitation are therefore needed, such as the opportunity to employ microbubbles to carry sonosensitizer or to employ new sensitizers specific for the SDT. Moreover, attention should be payed to the phenomenon of cavitation, which enables the sonochemical reactions to occur. Initiation of cavitation can be difficult, because of the high pressure required. Authors suggested some strategies to facilitate the cavitation, such as the application of standing waves rather than progressive or the dual frequency sonication [96]. i The last nRCT (identifier: NCT 04559685) is a Phase 0 single center, first in human, open-label study of ascend- ing energy doses of SDT utilizing the MRgFUS combined with intravenous ALA to assess safety and efficacy in up to 30 participants with recurrent HGG. The primary out- comes are to assess the biological changes associated with the SDT, analyzing the percentage of Cleaved Caspase-3, MIB-1 level and GammaH2Ax of the surgical specimen. Secondary outcomes include the evaluation of radiographic evidence of tumor physiological imaging changes and the assessment of performance, safety and tolerability of the MRgFUS and SDT. Ongoing clinical trials Other major concerns are that US procedures require long treatment sessions, therefore confining its application to small volumes, and the lack of in-vivo studies; in this regard efforts have been made and several ongoing clinical and pilot trials aim to better define the real clinical employ- ment of SDT in patients affected by GB [108]. Conclusions: challenges, limits, and future directions This review explored the current literature regarding the role of SDT in glioma treatment, and particularly in GB, con- sidering the evidence from ‘in vitro’ and ‘in vivo’ studies, and the ongoing clinical trials on its clinical human applica- tion. We also focused on the possible mechanisms of action underlying SDT and the role of different sonosensitizers. The study of the latter seems to enshrine the marriage of SDT and nanomedicine, paving the way for future research and new possibilities. f Challenges are also represented by the correct applica- tion into the neurooncological field of devices currently employed for other neurological disorders; an example is given by the essential tremor which benefit from the MRg- FUS performed with the ExAblate Neuro 4000. It is there- fore necessary to adapt and modify some characteristic such as the frequencies employed, which are lower in the setting of a tumor compared to the treatment of essential tremor (220 KHz vs 650 KHz) [109].At present, SDT is not Based on the studies that have been discussed on this paper and the current ongoing trials, SDT could be a 1 3 3 Journal of Neuro-Oncology (2023) 163:219–238 235 indicated as a first-line treatment for GB or any other type of brain cancer. the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. However, several scenarios are still open: SDT may have potential as an adjunctive treatment to enhance the effective- ness of standard therapies, such as chemotherapy via chemo- sensitization effect, or as a salvage option for patients who have failed other treatments, alone or in combination to other techniques to enhance its effect (e.g. hyperthermotherapy). In some cases, it may also be considered as a primary treat- ment strategy for patients who are not suitable candidates for surgery or who have recurrent tumors that are difficult to treat with other modalities.i References McMahon DJ, Gleeson JP, O’Reilly S, Bambury RM (2022) Management of newly diagnosed glioblastoma multiforme: cur- rent state of the art and emerging therapeutic approaches. Med Oncol 39:129. https://doi.org/10.1007/s12032-022-01708-w 7. McMahon DJ, Gleeson JP, O’Reilly S, Bambury RM (2022) Management of newly diagnosed glioblastoma multiforme: cur- rent state of the art and emerging therapeutic approaches. 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https://openalex.org/W3210468704	https://zenodo.org/records/3533790/files/63.a-energy%20drinks-converted.pdf	English	null	TRENDS OF ENERGY DRINKS, SOFT DRINKS AND BETEL NUTS CONSUMPTION IN MEDICAL UNIVERSITY STUDENTS: A USE OR AN ABUSE?	Zenodo (CERN European Organization for Nuclear Research)	2,019	cc-by	3,325	Nasreen Qazi et al Nasreen Qazi et al IAJPS 2019, 06 (11), 14392-14397 Nasreen Qa IAJPS 2019, 06 (11), 14392-14397 ISSN 2349-7750 Available online at: http://www.iajps.com Research Article TRENDS OF ENERGY DRINKS, SOFT DRINKS AND BETEL NUTS CONSUMPTION IN MEDICAL UNIVERSITY STUDENTS: A USE OR AN ABUSE? 1MBBS, Ph. D (Pharmacology), Professor and Chairperson, Department Of Pharmacology and Therapeutics, Liaquat University of Medical and Health Sciences, Jamshoro., 2MBBS, M. Phil (Pharmacology), Lecturer, Department Of Pharmacology and Therapeutics, LUMHS Jamshoro., 34th MBBS Student, Agha Khan University, Karachi, Pakistan., 4MBBS, MCPS (Family Medicine),M. Phil(Pharmacology), Consultant Family Physician, Assistant Professor Department of Pharmacology, Muhammad Medical College, Mirpurkhas, Sindh, Pakistan., 5MBBS, M. Phil (Biochemistry), Senior Lecturer, Department of Biochemistry, Isra University, Hyderabad, Pakistan., 6MBBS, FCPS (Medicine), Assistant Professor, Department of Medicine, Liaquat University of Medical and Health Sciences, Jamshoro. Article Received: September 2019 Accepted: October 2019 Published: November 2019 Article Received: September 2019 Accepted: October 2019 Published: November 2019 Abstract: Energy drinks, Soft Drinks consumption and Betel Nut chewing practices in students is a serious problem in developing countries. The aim of the present study was to determine the prevalence of energy drinks, soft drinks and betel nut consumption and their adverse effects in students of medical university. A questionnaire based cross sectional study was conducted on 316 students baes on questionnaire. We found 221 (69.94%) students consume energy drinks. Majority of them use energy drinks twice a week and most common brand is Sting. Students said that their energy level increased after taking energy drinks. 280 (88.6%) students consume soft drinks. Majority of them consume soft drink more than twice a week and were aware of the side effects. Out of 316 students, 136(43%) students chew betel nuts. Many of them consume betel nuts twice daily. Energy drinks, Soft Drinks consumption and Betel Nut chewing practices in students is a serious problem in developing countries. The aim of the present study was to determine the prevalence of energy drinks, soft drinks and betel nut consumption and their adverse effects in students of medical university. A questionnaire based cross sectional study was conducted on 316 students baes on questionnaire. We found 221 (69.94%) students consume energy drinks. Majority of them use energy drinks twice a week and most common brand is Sting. Students said that their energy level increased after taking energy drinks. 280 (88.6%) students consume soft drinks. Majority of them consume soft drink more than twice a week and were aware of the side effects. Out of 316 students, 136(43%) students chew betel nuts. Many of them consume betel nuts twice daily. Conclusion: The consumption of energy drinks, soft drinks and chewing betel nuts is a serious health problem and it is necessary to give awareness about the adverse effects of energy drinks, soft drinks and betel nuts to students. Keywords: Energy Drinks, Betel Nuts, Soft Drinks, Medical Students, Adverse Effects, Addiction. ( ) y f y Conclusion: The consumption of energy drinks, soft drinks and chewing betel nuts is a serious health problem and it is necessary to give awareness about the adverse effects of energy drinks, soft drinks and betel nuts to students. Keywords: Energy Drinks, Betel Nuts, Soft Drinks, Medical Students, Adverse Effects, Addiction. p g Prof. Dr. INTRODUCTION: MATERIALS AND METHODS: Energy drinks and soft drinks are the drinks which contains caffeine, taurine and glucose. It is considered that they increase the mental and physical energy levels. Energy drinks are different from sport drinks, sport drinks maintains the electrolytes balance and provide hydration during strenuous physical activity [1]. There are few concerns about safety profile of the usage of energy drinks and also they state that there are behavioral and physiological advantages in consumption of energy drinks [2]. Because of the increased concentration of caffeine in energy drinks, there are claims of certain side effects of these energy drinks on different body systems [3]. Most of the energy drinks contains high amount of glucose, taurine, riboflavin, pyridoxine, nicotinamide, vitamins B and many other stimulating herbal contents like guarana, ginseng and gingo biloba in addition of caffeine [4].Many energy drinks have approximately 80mg of caffeine which is equal to the amount of caffeine in 8 ounce of coffee cup [5]. Daily intake of 400mg caffeine is linked with many side effects like increase in anxiety, agitation, inability to sleep, ill temper, increased sweating, dehydration, nervousness, raised blood pressure and tachycardia [6].Energy drinks aims to target youngsters of age ranging from 18 to 35 years. Earlier researches have stated that students of universities and colleges are unaware of the harmful effects of energy drinks but they are not resistant to the harmful effects [7]. The consumption of soft drinks has drastically increased throughout the world in recent past [8]. The sugar sweetened beverages increase the risk of Weight Gain, Obesity, Type 2 Diabetes Mellitus, and disorders of cardiovascular system [9].These soft drinks are loaded with additional sugar Betel nut chewing is another injurious practice in university students. Betel nut chewing is one of the top five oral injurious habits across the globe and two hundred to six hundred million people chew betel nuts worldwide. It is the most common oral practice in India, srilanka, Taiwan and many more south Asian countries [10].The side effects of chewing betel nuts are mouth and pharyngeal cancer, oral premalignant lesions, conditions like oral leukoplakia and sub mucous fibrosis, diseases of gums and addictions. Abstract: Nasreen Qazi, Chairperson, Department of Pharmacology and therapeutics, LUMHS Email address: nk200684@gmail.com Please cite this article in press Nasreen Qazi et al., Trends Of Energy Drinks, Soft Drinks And Betel Nuts Consumption In Medical University Students: A Use Or An Abuse?., Indo Am. J. P. Sci, 2019; 06(11). Please cite this article in press Nasreen Qazi et al., Trends Of Energy Drinks, Soft Drinks And Betel Nuts Consumption In Medical University Students: A Use Or An Abuse?., Indo Am. J. P. Sci, 2019; 06(11). Page 14392 w w w . i a j p s . c o m w w w . i a j p s . c o m ISSN 2349-7750 Nasreen Qazi et al Nasreen Qazi et al IAJPS 2019, 06 (11), 14392-14397 INTRODUCTION: Chewing betel nuts is the fourth most prevalent addictive practice after tobacco, alcohol and caffeine across the globe [11].This study was designed to find out the consumption rate of energy drinks, soft drink and betel nuts and occurrence of their adverse effects among medical university students and whether they are aware about the side effects of energy drinks, soft drink and betel nuts. A cross sectional study was conducted in undergraduate students of MBBS by the Department of pharmacology in LUMHS Jamshoro in December 2017. Permission of data collection was sought from chair person of pharmacology department. Data was collected from 316 MBBS students. Sample was taken by simple random sampling technique. Sample size was calculated by using RAO SOFT sample size calculator. Oral informed consent of the student was taken. Data was collected by filling the questionnaire. Student were explained about how to fill up the questionnaire, Students were assured that all information would be kept confidential. Students were given a period of fifteen minutes to fill up the forms. Nasreen Qazi et al Nasreen Qazi et al Nasreen Qazi et al ISSN 2349-7750 ISSN 2349-7750 IAJPS 2019, 06 (11), 14392-14397 RESULTS: The data was collected from 316 MBBS students, selected by simple random sampling technique. Out of 316 students 198 (62.66%) students were male and 118 (37.34%) students were female (Graph: 1 showing Gender Distribution).From the 316 MBBS students who participated in our study 221 (69.94%) students were using energy drinks while 95 (30.06%) students replied that they don’t use energy drinks. Majority of students 140 (63.3%) take energy drink’s brand “Sting” and 81 (36.7%) students consume energy drink’s brand “Red Bul”. 75 (33.9%) students consumed energy drinks once a week 79 (35.7%) students consumed energy drinks twice a weeks and 67 (30.4%) students used energy drinks daily. 110 (49.8%) students said that they feel their energy level increased after taking energy drinks, 60 (27.1%) students experienced increased alertness 30 (13.6%) students experienced increased in heart rate 10 (45.5%) students said that their blood pressure increased after taking energy drinks and 11 (5%) students have said that they don’t experience any effect after taking energy drinks. When students were asked why they take energy drinks, most of them 118 (53.4%) said that they take energy drinks to increase the mental performance and 103 (46.6%) said that they take energy drinks just for fun. From 316 students 280 (88.6%) students said that they take soft drinks. 36 (11.4%) students said that they don’t use soft drinks. Majority of the students 170 (60.7%) said that they consumed brand “Pepsi”, 81 (28.9%) students consumed “7-Up” and 29 (10.4%) students consumed “Miranda”. 38 (13.6%) students take soft drinks once a week. 121 (43.2%) students take soft drinks twice a weeks and 121 (43.2) students take soft drinks daily. 131 (40.4%) students take soft drinks for rehydration. 52 (18.6%) students take soft drinks to reduce fatigue and 115 (41 %) students take soft drinks for taste. Out of 316 students 249 (78.8%) students were aware of Page 14393 w w w . i a j p s . c o m IAJPS 2019, 06 (11), 14392-14397 that they consumed betel nuts twice daily and 45 (33.1%) students said that they consumed betel nuts more than twice daily. 278 (88%) students were aware of the side effects of betel nuts 38 (12%) students said that they are not aware of the side effects of betel nuts. the side effects of soft drinks and 67 (21.2%) students reported that they are unaware of the side effects of the soft drinks.Out of 316 students 136 (43%) students take betel nuts and 180 (57%) said that they don’t consume betel nuts. 37 (25%) students said that they consumed betel nuts daily. 57 (41.9%) students said Figure.1: Gender Distribution represented by pie chart. 62.64% 37.36% GenderDistribution Males Females Figure.1: Gender Distribution represented by pie chart. 62.64% 37.36% GenderDistribution Males Females Figure.1: Gender Distribution represented by pie chart. Table-1: Frequency and percentage of Betel nuts Consumption Variables Frequency Percent Students taking Betel Nuts Yes 136 43 No 180 57 Consumption of Betel Nuts / Day once daily 34 25 twice Daily 57 41.9 More than twice daily 45 33.1 Awareness about the Side Effects of Betel Nuts Yes 278 88 No 38 12 62.64% 37.36% GenderDistribution Males Females GenderDistribution Figure.1: Gender Distribution represented by pie chart. Figure.1: Gender Distribution represented by pie chart. Table-1: Frequency and percentage of Betel nuts Consumption Variables Frequency Percent Students taking Betel Nuts Yes 136 43 No 180 57 Consumption of Betel Nuts / Day once daily 34 25 twice Daily 57 41.9 More than twice daily 45 33.1 Awareness about the Side Effects of Betel Nuts Yes 278 88 No 38 12 Page 14394 w w w . i a j p s . c o m Nasreen Qazi et al Nasreen Qazi et al ISSN 2349-7750 J , ( ), Q i a j p s . c o m Page 143 Table: 1. Frequency and percentage of Energy and Soft Drinks Consumption. DISCUSSION: there is a need to educate the undergraduate medical students about the potential adverse effects of energy drinks, soft drinks and betel nuts. They consumption of energy drinks and soft drinks is increasing in young generation specifically in college and university students. Majority of the students are using energy drinks and soft drinks now a days and the main reason for the increase consumption of these caffeinated drinks is their easy availability in educational institutes and because these drinks are marketed as they are very beneficial for one’s health and mental performance. This study was conducted in December 2017 to assess the frequency of consumption of soft drinks, energy drinks and betel nuts. Which brand of these drinks are in common use, their effects and whether students are aware of the adverse effects of these drinks and betel nuts. Approximately more than half of the students, who took part in our study, were consuming soft drinks. A previous study Denney-Wilson et al was in agreement with our study that the majority of the students consumed soft drinks [12]. This study also stated that one of the main reasons of increased consumption of soft drinks is because of their taste. TERRY-McElrath et al previously stated that approximately half of the students of their study sample consumed soft drinks daily which was in contrast to our study [13]. In our study approximately one third of the students consumed soft drinks daily. A previous study was also in agreement with our study that students take caffeinated drinks for rehydration. However usage of these drinks for rehydration after physical activity is dangerous for health [14]. In our study total 221(69.94%) students from 316 students consume energy drinks. Majority of them consume energy drinks to increase energy level and to increase alertness. A previous study was in agreement to our study [14]. Another previous study was in contrast to our study. They have lower proportion of students who consume energy drinks than our study [15]. In our study majority of students 140 (63.3%), consumed energy drink’s brand “sting” because of its taste, easy availability and low price which in accordance with Saeed Ahmed et al [16]. Previous studies Aljaloud SO et al[17]and usman et al[18] were in contrast to our study that majority of their students consume energy drink’s brand “red bul”. DISCUSSION: A previous study stated that majority of students consume energy drinks for enjoyment and to increase mental performance [19]. A previous study also stated that very few students were aware of the potential adverse effects of betel nut chewing which is in contrast to our study [20]. Our study is stating that majority of students are aware of the side effects of betel nut consumption. Another study was in agreement to our study that many students of their sample size were aware of the side effects of betel nut consumption [21]. In our opinion CONCLUSION: Majority of undergraduate medical students consume energy drinks and soft drinks and many of the students chew betel nuts despite knowing of their side effects, they still consume them. Students believe that energy drinks and soft drinks increase their mental alertness and give them energy. IAJPS 2019, 06 (11), 14392-14397 n=316 VARIABLES ENERGY DRINKS SOFT DRINKS STUDENTS USING DRINKS Yes 221(69.94%) 280(88.6%) No 95(30.06%) 36(11.4%) BRAND OF DRINKS Sting 140(63.3%) Pepsi 170(60.7%) Red Bull 81(36.7%) 7-up 81(28.9%) --------------------------------- ------------------------ Marinda 29(10.4%) CONSUMPTION OF DRINKS PER WEEK Once a week 75(33.9%) 38(13.6%) Twice a week 79(35.7%) 121(43.2%) More than twice a week 67(30.4%) 121(43.2%) AWARENESS ABOUT SIDE EFFECTS OF DRINKS Yes 305(95%) 249(78.8%) No 11(5%) 67(21.2%) REASONS TO TAKE ENERGY DRINKS To increase mental alertness 118(5.4%) 52(18.6%) For Fun 103(46.6%) 115(41%) Rehydration --------------- 131(40.4%) Fig.2: Bar charts of frequency of consumption of energy drinks, soft drinks and betel nuts 0 50 100 150 200 250 300 Energy Drinks Soft Drinks Betal Nuts 221 280 136 95 36 180 Yes No Table: 1. Frequency and percentage of Energy and Soft Drinks Consumption. n=316 VARIABLES ENERGY DRINKS SOFT DRINKS STUDENTS USING DRINKS Yes 221(69.94%) 280(88.6%) No 95(30.06%) 36(11.4%) BRAND OF DRINKS Sting 140(63.3%) Pepsi 170(60.7%) Red Bull 81(36.7%) 7-up 81(28.9%) --------------------------------- ------------------------ Marinda 29(10.4%) CONSUMPTION OF DRINKS PER WEEK Once a week 75(33.9%) 38(13.6%) Twice a week 79(35.7%) 121(43.2%) More than twice a week 67(30.4%) 121(43.2%) AWARENESS ABOUT SIDE EFFECTS OF DRINKS Yes 305(95%) 249(78.8%) No 11(5%) 67(21.2%) REASONS TO TAKE ENERGY DRINKS To increase mental alertness 118(5.4%) 52(18.6%) For Fun 103(46.6%) 115(41%) Rehydration --------------- 131(40.4%) Fig.2: Bar charts of frequency of consumption of energy drinks, soft drinks and betel nuts 0 50 100 150 200 250 300 Energy Drinks Soft Drinks Betal Nuts 221 280 136 95 36 180 Yes No Table: 1. Frequency and percentage of Energy and Soft Drinks Consumption. n=316 Fig.2: Bar charts of frequency of consumption of energy drinks, soft drinks and betel nuts 0 50 100 150 200 250 300 Energy Drinks Soft Drinks Betal Nuts 221 280 136 95 36 180 Yes No Fig.2: Bar charts of frequency of consumption of energy drinks, soft drinks and betel nuts Page 14395 w w w . i a j p s . c o m IAJPS 2019, 06 (11), 14392-14397 Nasreen Qazi et al ISSN 2349-7750 ISSN 2349-7750 REFERENCES: REFERENCES: 1. Andrew P Smith, Gareth Richards92018). Energy drinks, caffeine, junk food, breakfast, depression and academic attainment of secondary school students. Journal of Psychopharmacology.32(8) 893– 899. 2. Alsunni AA, Badar A(2011). Energy drinks consumption pattern, perceived benefits and associated adverse effects amongst students of university of Dammam, Saudi Arabia. J Ayub MedColl Abbottabad 23:3‑9. 3. Arria AM, Bugbee BA, Caldeira KM, Vincent KB(2014). 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https://openalex.org/W3171147107	https://www.medrxiv.org/content/medrxiv/early/2021/06/04/2021.06.01.21258117.full.pdf	English	null	<i>Plasmodium falciparum hrp2</i> and <i>hrp3</i> gene deletion status in Africa and South America by highly sensitive and specific digital PCR	medRxiv (Cold Spring Harbor Laboratory)	2,021	cc-by	20,610	. CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: nt . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Plasmodium falciparum hrp2 and hrp3 gene deletion status in Africa and South America by 1 highly sensitive and specific digital PCR 2 3 Short title: hrp2/hrp3 deletion typing by digital PCR 4 5 Claudia A. Vera-Arias1, Aurel Holzschuh1,2, Colins O. Oduma3,4, Kingsley Badu5, Mutala Abdul-Hakim5, 6 Joshua Yukich6, Manuel W. Hetzel2,7, Bakar S. Fakih2,7,8, Abdullah Ali9, Marcelo U. Ferreira10, Simone 7 Ladeia-Andrade11, Fabián E. Sáenz12, Yaw Afrane13, Endalew Zemene14, Delenasaw Yewhalaw14, James W. 8 Kazura15, Guiyun Yan16, Cristian Koepfli1 9 10 1 University of Notre Dame, Notre Dame, IN, United States 11 2 Swiss Tropical and Public Health Institute, Switzerland 12 3 Kenya Medical Research Institute-Centre for Global Health Research, Kisumu, Kenya 13 4 Department of Biochemistry and Molecular Biology, Egerton University, Nakuru, Kenya 14 5 Kwame Nkrumah University of Science and Technology, Ghana 15 6 Tulane University, USA 16 7 University of Basel, Basel, Switzerland 17 8 Ifakara Health Institute, Dar es Salaam, United Republic of Tanzania 18 9 Zanzibar Malaria Elimination Programme, Zanzibar, United Republic of Tanzania 19 10 University of Sao Paulo, Brazil 20 11 Laboratory of Parasitic Diseases, Fiocruz, Rio de Janeiro, Brazil 21 12 Centro de Investigación para la Salud en América Latina, Facultad de Ciencias Exactas y Naturales, 22 Pontificia Universidad Católica del Ecuador, Quito, Ecuador 23 Plasmodium falciparum hrp2 and hrp3 gene deletion status in Africa and South America by 1 highly sensitive and specific digital PCR 2 13 Department of Medical Microbiology, University of Ghana Medical School, Accra, Ghana 24 NOTE: This preprint reports new research that has not been certified by peer review and should not be used to guide clinical practice. . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint 14 Tropical and Infectious Diseases Research Center, Jimma University, Ethiopia 25 15 Case Western Reserve University, Cleveland, USA 26 16 Program in Public Health, University of California, Irvine, CA 92617, USA 27 28 * Corresponding author: ckoepfli@nd.edu 29 30 31 32 32 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. h was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: iv preprint Abstract 33 34 Background: The most commonly used Plasmodium falciparum rapid diagnostic tests target the Histidine- 35 Rich Proteins 2 and 3 (HRP2, HRP3). An increasing number of countries report parasites that carry hrp2 36 and/or hrp3 gene deletions, resulting in false negative test results. Molecular surveillance of hrp2 and 37 hrp3 deletions is crucial but adequate protocols have been lacking. Abstract 33 Methods and Findings: We have developed novel assays for deletion typing based on droplet digital PCR 39 (ddPCR), targeting hrp2 exon1, hrp2 exon 2, and hrp3. In the ddPCR assay, hrp2 or hrp3 and a control gene 40 were quantified with very high accuracy in a single tube. The theoretical limit of detection of the ddPCR 41 assay was 0.33 parasites/uL, and thus well suited for typing of low-density asymptomatic infections. The 42 deletion was reliably detected in mixed infections with wild-type and hrp2-deleted parasites when the 43 proportion of parasites carrying the deletion was >40%. For a side-by-side comparison with the 44 conventional nested PCR (nPCR) assay, 248 samples from asymptomatic individuals from western Kenya 45 were screened in triplicate by ddPCR and nPCR. No deletions were observed by ddPCR, while by nPCR no 46 band for hrp2 was observed in 8% of samples. The ddPCR assay was applied to screen 830 samples from 47 six countries in Africa and South America. No or very few deletions were observed in Kenya (n=241), 48 Zanzibar/Tanzania (n=91), and Ghana (n=223). In southwestern Ethiopia, 1/47 (2.1%) samples carried hrp2 49 deletion, and 35/47 (74.5%) hrp3 deletions. In Brazil, 87/187 (46.5%) samples carried hrp2 deletions, and 50 116/187 (62%) hrp3 deletions. In Ecuador, no hrp2 deletions were observed, but 22/41 (53.7%) samples 51 carried hrp3 deletions. 52 Methods and Findings: We have developed novel assays for deletion typing based on droplet digital PCR 39 (ddPCR), targeting hrp2 exon1, hrp2 exon 2, and hrp3. In the ddPCR assay, hrp2 or hrp3 and a control gene 40 were quantified with very high accuracy in a single tube. The theoretical limit of detection of the ddPCR 41 assay was 0.33 parasites/uL, and thus well suited for typing of low-density asymptomatic infections. The 42 deletion was reliably detected in mixed infections with wild-type and hrp2-deleted parasites when the 43 proportion of parasites carrying the deletion was >40%. For a side-by-side comparison with the 44 conventional nested PCR (nPCR) assay, 248 samples from asymptomatic individuals from western Kenya 45 were screened in triplicate by ddPCR and nPCR. No deletions were observed by ddPCR, while by nPCR no 46 band for hrp2 was observed in 8% of samples. The ddPCR assay was applied to screen 830 samples from 47 six countries in Africa and South America. Plasmodium falciparum hrp2 and hrp3 gene deletion status in Africa and South America by 1 highly sensitive and specific digital PCR 2 38 Abstract 33 34 Background: The most commonly used Plasmodium falciparum rapid diagnostic tests target the Histidine- 35 Rich Proteins 2 and 3 (HRP2, HRP3). An increasing number of countries report parasites that carry hrp2 36 and/or hrp3 gene deletions, resulting in false negative test results. Molecular surveillance of hrp2 and 37 hrp3 deletions is crucial but adequate protocols have been lacking. 38 Methods and Findings: We have developed novel assays for deletion typing based on droplet digital PCR 39 (ddPCR), targeting hrp2 exon1, hrp2 exon 2, and hrp3. In the ddPCR assay, hrp2 or hrp3 and a control gene 40 were quantified with very high accuracy in a single tube. The theoretical limit of detection of the ddPCR 41 assay was 0.33 parasites/uL, and thus well suited for typing of low-density asymptomatic infections. The 42 deletion was reliably detected in mixed infections with wild-type and hrp2-deleted parasites when the 43 proportion of parasites carrying the deletion was >40%. For a side-by-side comparison with the 44 conventional nested PCR (nPCR) assay, 248 samples from asymptomatic individuals from western Kenya 45 were screened in triplicate by ddPCR and nPCR. No deletions were observed by ddPCR, while by nPCR no 46 band for hrp2 was observed in 8% of samples. The ddPCR assay was applied to screen 830 samples from 47 six countries in Africa and South America. No or very few deletions were observed in Kenya (n=241), 48 Zanzibar/Tanzania (n=91), and Ghana (n=223). In southwestern Ethiopia, 1/47 (2.1%) samples carried hrp2 49 deletion, and 35/47 (74.5%) hrp3 deletions. In Brazil, 87/187 (46.5%) samples carried hrp2 deletions, and 50 116/187 (62%) hrp3 deletions. In Ecuador, no hrp2 deletions were observed, but 22/41 (53.7%) samples 51 carried hrp3 deletions. 52 Conclusions: Compared to nPCR, the ddPCR assay minimizes the risk of false-negative results (i.e. hrp2 53 deletion observed when the sample is wild type), increases sensitivity, and greatly reduces the number of 54 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Abstract 33 No or very few deletions were observed in Kenya (n=241), 48 Zanzibar/Tanzania (n=91), and Ghana (n=223). In southwestern Ethiopia, 1/47 (2.1%) samples carried hrp2 49 deletion, and 35/47 (74.5%) hrp3 deletions. In Brazil, 87/187 (46.5%) samples carried hrp2 deletions, and 50 116/187 (62%) hrp3 deletions. In Ecuador, no hrp2 deletions were observed, but 22/41 (53.7%) samples 51 carried hrp3 deletions. 52 Conclusions: Compared to nPCR, the ddPCR assay minimizes the risk of false-negative results (i.e. hrp2 53 deletion observed when the sample is wild type), increases sensitivity, and greatly reduces the number of 54 reactions that need to be run. Pronounced differences in the prevalence of deletion were observed among 55 sites, with more hrp3 than hrp2 deletions. 56 Conclusions: Compared to nPCR, the ddPCR assay minimizes the risk of false-negative results (i.e. hrp2 53 deletion observed when the sample is wild type), increases sensitivity, and greatly reduces the number of 54 reactions that need to be run. Pronounced differences in the prevalence of deletion were observed among 55 sites, with more hrp3 than hrp2 deletions. 56 57 58 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. h was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: iv preprint Introduction 59 60 In 2019, over 200 million cases of malaria and over 400,000 deaths were recorded [1]. Plasmodium 61 falciparum remains the primary cause of malaria in humans. Fast and accurate diagnosis and treatment 62 of clinical episodes are key components of malaria control. Diagnosis is commonly performed either by 63 light microscopy, or rapid diagnostic tests (RDTs). RDTs are lateral flow devices that detect parasite 64 proteins in human blood through immunohistochemistry. Light microscopy requires basic lab 65 infrastructure and skilled microscopists. In contrast, RDTs require minimal infrastructure and training, and 66 results are available within approximately 15 minutes at a cost of less than 1 USD per test. Abstract 33 RDTs are the 67 only field-deployable diagnostic tool available at peripheral health centers and for community screening 68 to diagnose asymptomatic infections, for example through reactive case detection [2]. In 2016, over 300 69 million RDTs were used by malaria control programs [3]. 70 Introduction 59 60 In 2019, over 200 million cases of malaria and over 400,000 deaths were recorded [1]. Plasmodium 61 falciparum remains the primary cause of malaria in humans. Fast and accurate diagnosis and treatment 62 of clinical episodes are key components of malaria control. Diagnosis is commonly performed either by 63 light microscopy, or rapid diagnostic tests (RDTs). RDTs are lateral flow devices that detect parasite 64 proteins in human blood through immunohistochemistry. Light microscopy requires basic lab 65 infrastructure and skilled microscopists. In contrast, RDTs require minimal infrastructure and training, and 66 results are available within approximately 15 minutes at a cost of less than 1 USD per test. RDTs are the 67 only field-deployable diagnostic tool available at peripheral health centers and for community screening 68 to diagnose asymptomatic infections, for example through reactive case detection [2]. In 2016, over 300 69 million RDTs were used by malaria control programs [3]. 70 Introduction 59 Introduction 59 60 In 2019, over 200 million cases of malaria and over 400,000 deaths were recorded [1]. Plasmodium 61 falciparum remains the primary cause of malaria in humans. Fast and accurate diagnosis and treatment 62 of clinical episodes are key components of malaria control. Diagnosis is commonly performed either by 63 light microscopy, or rapid diagnostic tests (RDTs). RDTs are lateral flow devices that detect parasite 64 proteins in human blood through immunohistochemistry. Light microscopy requires basic lab 65 infrastructure and skilled microscopists. In contrast, RDTs require minimal infrastructure and training, and 66 results are available within approximately 15 minutes at a cost of less than 1 USD per test. RDTs are the 67 only field-deployable diagnostic tool available at peripheral health centers and for community screening 68 to diagnose asymptomatic infections, for example through reactive case detection [2]. In 2016, over 300 69 million RDTs were used by malaria control programs [3]. 70 71 The most sensitive RDTs for P. falciparum rely on the detection of Histidine Rich Protein 2 (HRP2) [4]. HRP2 72 is a highly expressed secreted protein, and thus an ideal target for diagnosis. It is also the target for a new 73 generation of ultra-sensitive RDTs with a limit of detection of <100 parasites/µL [5, 6]. While alternative 74 RDTs detecting other proteins, e.g. parasite lactase dehydrogenase (pLDH), or aldolase, are available, they 75 are generally less sensitive [7, 8]. HRP2-based RDTs can also detect HRP3, a structurally similar protein 76 sharing multiple epitopes with HRP2. 77 78 In 2010, a report revealed that a large proportion of P. falciparum parasites in Peru did not carry the hrp2 79 gene [9], and thus could not be detected by HRP2-based RDTs. Since then, an increasing number of reports 80 from Latin America [10-14], Africa [15-22], and Asia [23, 24] found varying proportions of parasites with 81 hrp2-deletion, reaching up to 80% of clinical cases in certain hospitals in Eritrea [18]. In addition, hrp3 can 82 be deleted. The deletion of either hrp2 or hrp3, or both genes, has no known impact on parasite fitness. 83 RDTs can yield a positive result if hrp2 is deleted but hrp3 is expressed, but sensitivity of the RDT is lower 84 in this case [25]. Introduction 59 85 86 Molecular surveillance to assess the frequency of hrp2 and hrp3 deletion is crucial to decide whether 87 HRP2-based RDTs can be used [26]. The WHO recommends to use alternative diagnostics if the prevalence 88 of hrp2 deletion is above 5% [27]. At this level, the number of false-negative tests due to hrp2 deletion 89 will exceed the number of false-negative tests because of lower sensitivity of the alternative diagnostics. 90 The prevalence of hrp2 deletion has been found to differ substantially within countries [12, 17], thus the 91 choice of diagnostics might need to be adapted at sub-national level. Where low levels of deletions are 92 present, HRP2-based RDTs remain a highly useful tool for diagnosis. 93 94 Deletion screening has been classically done using nested PCR (nPCR) followed by gel electrophoresis [26]. 95 Absence of a band is interpreted as deletion. False-negative results could occur when PCR conditions are 96 suboptimal, or when parasite density is low and amplification is stochastic. To overcome this limitation, 97 3-fold repetition of the nested hrp2 PCR and a control PCR (e.g. msp2, or glurp) is recommended [26]. As 98 a result, for each sample twelve PCRs need to be run. Deletion status might remain unresolved if results 99 differ among replicates. As an additional problem, multiple clone P. falciparum infections are common in 100 most endemic settings [28]. In case of a multiple clone infection with a wild-type parasite and a parasite 101 carrying the deletion, the wild type parasite will result in a band on the gel when using the nPCR assay. 102 Multiple clone infections can thus mask the presence of deletions, resulting in an underestimation of the 103 frequency of deletion [29]. More recently, quantitative PCR (qPCR) protocols for hrp2/hrp3 deletion typing 104 were published [30], greatly enhancing throughput. However, when parasite densities are low, 105 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. Introduction 59 (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint be deleted. The deletion of either hrp2 or hrp3, or both genes, has no known impact on parasite fitness. 83 RDTs can yield a positive result if hrp2 is deleted but hrp3 is expressed, but sensitivity of the RDT is lower 84 in this case [25]. 85 86 Molecular surveillance to assess the frequency of hrp2 and hrp3 deletion is crucial to decide whether 87 HRP2-based RDTs can be used [26]. The WHO recommends to use alternative diagnostics if the prevalence 88 of hrp2 deletion is above 5% [27]. At this level, the number of false-negative tests due to hrp2 deletion 89 will exceed the number of false-negative tests because of lower sensitivity of the alternative diagnostics. 90 The prevalence of hrp2 deletion has been found to differ substantially within countries [12, 17], thus the 91 choice of diagnostics might need to be adapted at sub-national level. Where low levels of deletions are 92 present, HRP2-based RDTs remain a highly useful tool for diagnosis. 93 94 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint considerable variation in quantification is observed between replicates [31]. Introduction 59 As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 108 We have developed a novel method for typing of hrp2 and hrp3 deletions based on droplet digital PCR 109 (ddPCR). ddPCR yields highly accurate quantification of parasites [31]. In a ddPCR experiment, the reaction 110 volume is partitioned into approximately 15,000 microdroplets, which are then subject to end-point PCR. 111 Each droplet functions as an individual PCR reaction, with amplification occurring if the droplet contains 112 template DNA. The number of positive droplets corresponds to the number of template DNA copies. Using 113 two different probes, two targets can be quantified in a single reaction well, e.g. a control gene and a 114 target gene. As each droplet with template can be considered a ‘within-well replicate’, a single well offers 115 the sensitivity and specificity of a large number of replicates by nPCR or qPCR. The risk of false negative 116 results (i.e. no amplification when template is present) is thus minimal compared to nPCR or qPCR. The 117 novel assay greatly reduces the number of reactions to be run, increase sensitivity and accuracy, and can 118 detect the deletion in mixed infections. The assay was extensively validated using culture strains, and field 119 isolates from Kenya, Zanzibar/Tanzania, Ethiopia, Ghana, Brazil, and Ecuador. 120 121 considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 The study was approved by the University of Notre Dame Institutional Review Board 129 (approvals 18-08-4803, 19-04-5321, 18-12-5029), the Institutional Scientific and Ethical Review boards of 130 the Noguchi Memorial Institute of Medical Research, University of Ghana, the Committee on Human 131 Research, Publication and Ethics, School of Medical Science, Kwame Nkrumah University of Science and 132 Technology, Kumasi (CHRPE/AP/375/20), the Zanzibar Medical Research Ethics Committee 133 (ZAMREC/0001/Feb/17), the Institutional Review Board of Tulane University (17-993573), the Institutional 134 Review Board of the Ifakara Health Institute (003-2017), the Ethics Commission of North-western and 135 Central Switzerland (Req-2017-00162), the Institutional Review Board of Institute of Health, Jimma 136 University, Ethiopia (RPGC/486/06), Maseno University Ethics Review Committee (MUERC protocol 137 number 00456), the Ethics Committee for Research in Human Beings of the Pontificia Universidad Católica 138 del Ecuador (CEISH-571-2018), the Ministry of Public Health of Ecuador (MSP-DIS-2019-004-O), and the 139 institutional review board of Oswaldo Cruz Foundation, Brazil (no. 022/2009). 140 126 Ethics statement 127 Informed written consent was obtained from all study participants or their parents or legal guardians prior 128 to sample collection. The study was approved by the University of Notre Dame Institutional Review Board 129 (approvals 18-08-4803, 19-04-5321, 18-12-5029), the Institutional Scientific and Ethical Review boards of 130 the Noguchi Memorial Institute of Medical Research, University of Ghana, the Committee on Human 131 Research, Publication and Ethics, School of Medical Science, Kwame Nkrumah University of Science and 132 Technology, Kumasi (CHRPE/AP/375/20), the Zanzibar Medical Research Ethics Committee 133 (ZAMREC/0001/Feb/17), the Institutional Review Board of Tulane University (17-993573), the Institutional 134 Review Board of the Ifakara Health Institute (003-2017), the Ethics Commission of North-western and 135 Central Switzerland (Req-2017-00162), the Institutional Review Board of Institute of Health, Jimma 136 University, Ethiopia (RPGC/486/06), Maseno University Ethics Review Committee (MUERC protocol 137 number 00456), the Ethics Committee for Research in Human Beings of the Pontificia Universidad Católica 138 del Ecuador (CEISH-571-2018), the Ministry of Public Health of Ecuador (MSP-DIS-2019-004-O), and the 139 institutional review board of Oswaldo Cruz Foundation, Brazil (no. 022/2009). 140 141 Digital PCR assays 142 Four novel ddPCR assays were developed. One assay targets the conserved first 120 bp of hrp2 exon 2, 143 and thus is located directly adjacent to the histidine-rich repeats. Different breakpoint for the hrp2 144 deletion have been described [26]. . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Methods 125 126 Ethics statement 127 Informed written consent was obtained from all study participants or their parents or legal guardians prior 128 to sample collection. The study was approved by the University of Notre Dame Institutional Review Board 129 (approvals 18-08-4803, 19-04-5321, 18-12-5029), the Institutional Scientific and Ethical Review boards of 130 the Noguchi Memorial Institute of Medical Research, University of Ghana, the Committee on Human 131 Research, Publication and Ethics, School of Medical Science, Kwame Nkrumah University of Science and 132 Technology, Kumasi (CHRPE/AP/375/20), the Zanzibar Medical Research Ethics Committee 133 (ZAMREC/0001/Feb/17), the Institutional Review Board of Tulane University (17-993573), the Institutional 134 Review Board of the Ifakara Health Institute (003-2017), the Ethics Commission of North-western and 135 Central Switzerland (Req-2017-00162), the Institutional Review Board of Institute of Health, Jimma 136 University, Ethiopia (RPGC/486/06), Maseno University Ethics Review Committee (MUERC protocol 137 number 00456), the Ethics Committee for Research in Human Beings of the Pontificia Universidad Católica 138 del Ecuador (CEISH-571-2018), the Ministry of Public Health of Ecuador (MSP-DIS-2019-004-O), and the 139 institutional review board of Oswaldo Cruz Foundation, Brazil (no. 022/2009). 140 141 Methods 125 126 Ethics statement 127 Informed written consent was obtained from all study participants or their parents or legal guardians prior 128 to sample collection. considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 The novel primers for exon 2 are located in a region that is deleted in 145 all known deletion variants, thus irrespective of the specific breakpoint, hrp2 deletion will be detected. 146 The second assay targets hrp2 exon 1. While exon 1 does not contain antigens that are recognized by 147 RDTs, the deletion of this exon 1 would prevent proper expression of the protein. The third assay targets 148 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint hrp3. Each assay was multiplexed with an assay targeting serine-tRNA ligase (PF3D7_0717700, herein 149 referred to as ‘tRNA’). tRNA is a conserved, essential single copy gene, that is frequently used as reference 150 for gene expression assays [32]. In wild type infections not carrying a deletion, the copy numbers of tRNA 151 and hrp2 or hrp3 are identical. 152 153 Novel primers and probes were developed for all assays (Table 1). Assay conditions are given in 154 Supplementary File S1. Across over >3000 genomes available through MalariaGen and PlasmoDB, no SNPs 155 were recorded in primer and probe sequences, thus the assay can be used for the screening of isolates of 156 global origin. Assay conditions were optimized to achieve maximal separation between positive and 157 negative droplets (Figure 1, Supplementary File S2, Figure S1). 158 159 Table 1: Primer and probe sequences 160 161 Assay Sequence 5'-3' hrp2 exon 2 forward CATTTTTAAATGCTTTTTTATTTTTATATAG hrp2 exon 2 hrp2 exon 2 reverse CTTGAGTTTCGTGTAATAATCTC hrp2 exon 2 probe FAM-CGCATTTAATAATAACTTGTGTAGCAAAAATGC hrp2 exon 1 forward ATATTTATACATTTTTGTTATTATTTCTTTTTC hrp2 exon 1 hrp2 exon 1 reverse CGTTATCTAACAAAAGTACGGAG hrp2 exon 1 probe FAM-CAAAAACGGCAGCGGATAATACTT hrp3 forward ATGCTAATCACGGATTTCATTTTA hrp3 hrp3 reverse ATCGTCATGGTGAGAATCATC hrp3 probe FAM-CCTTCACGATAACAATTCCCATACTTTAC tRNA forward CATCAAATGAAGATTTAACAAGAG tRNA tRNA reverse CTTTTTGATTCTATAGTTTCATCTTTATG tRNA probe HEX-CTACCTCAGAACAACCATTATGTGCT hrp3. Each assay was multiplexed with an assay targeting serine-tRNA ligase (PF3D7_0717700, herein 149 referred to as ‘tRNA’). considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 tRNA is a conserved, essential single copy gene, that is frequently used as reference 150 for gene expression assays [32]. In wild type infections not carrying a deletion, the copy numbers of tRNA 151 and hrp2 or hrp3 are identical. 152 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: iv preprint 162 3D7 and Dd2 parasite culture strain mixtures 163 In order to determine the ability to detect mixed clone infections with only one strain carrying the 164 deletion, experimental mixtures were made from two well-characterized laboratory strains, 3D7 (no 165 deletions), and Dd2 (carrying the hrp2 deletion). Each strain was cultured separately, DNA extracted, and 166 quantified by ddPCR using the hrp2/tRNA ligase assay. No hrp2 was detected in Dd2. Mixtures were 167 prepared with a concentration (of both strains combined) of 10, 50, 100, and 500 genomes/µL, and with 168 a 3D7 to Dd2 ratio of 100:0, 80:20, 60:40, 40:60, 20:80, and 0:100. Each mixture was run in triplicate using 169 the hrp2 exon2/ tRNA ligase assay. The ratio of hrp2 to tRNA copy numbers was compared to expected 170 values. 171 172 Nested PCR assay for hrp2 173 In order to compare the ddPCR assay directly to the established nPCR assay, 248 samples from 174 asymptomatic carriers in western Kenya were run by the hrp2 exon 2 assay, and the hrp2 nsPCR in 175 triplicate. Assay conditions for the nPCR followed published protocols [33, 34] and are given in 176 Supplementary File S1. 177 178 Field isolates 179 Field isolates were screened from Kenya, Zanzibar in the United Republic of Tanzania, Ethiopia, Ghana, 180 Brazil, and Ecuador. Samples from Kenya (n=248) were from Chulaimbo and Homa Bay in western Kenya 181 close to Lake Victoria. Samples were collected in a cross-sectional survey including individuals of all ages 182 from January-August 2019. considerable variation in quantification is observed between replicates [31]. As a result of the technical 106 challenges for accurate typing, maps of hrp2 deletion frequency remain scattered and incomplete. 107 Finger-prick samples were collected in EDTA microtainers and infections 183 detected by qPCR. Overall P. falciparum prevalence across both sites was 16% [35]. No diagnosis by RDT 184 or microscopy was done. 185 3D7 and Dd2 parasite culture strain mixtures 163 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint 186 Samples from Zanzibar (n=91) had been collected in the frame of a study on reactive case detection (RCD) 187 from May 2017 to October 2018 [2]. Asymptomatic infections identified through RCD were typed. During 188 RCD, infections were diagnosed by HRP2/pLDH-based RDT (SD BIOLINE Malaria Ag Pf HRP2/pLHD), a blood 189 spot was collected on filter paper, and infections diagnosed by qPCR [2]. Prevalence (not including index 190 cases) was 0.8% by RDT and 2.4% by qPCR. All samples with a density (by qPCR) of >100 parasites/µL were 191 selected for hrp2/hrp3 deletion typing, irrespective of RDT result. 192 193 Samples from Ethiopia (n=47) included clinical cases and asymptomatic individuals sampled in June to 194 November 2016 from Jimma Zone, Oromia Region. P. falciparum prevalence was 4% by microscopy and 195 8.3% by qPCR [36]. No RDT screening was conducted. 196 197 From Ghana, 2 sets of samples were typed. The first set (n=11) was collected in June-September of 2017 198 from febrile school children aged 5-14 years [37]. The second set (n=212) was collected in Mankranso and 199 Agona Hospitals in the Ashanti region from febrile patients from September to December 2020. 200 201 In Brazil, samples (n=187) were collected in Cruzeiro do Sul, Upper Juruá Valley, northwestern Brazil, in 202 2010-2013. This is the country¢s main malaria hotspot, which accounted for nearly 15% of all P. falciparum 203 infections in Brazil at the time of the study. Samples were collected from clinical patients 4 to 73 years of 204 age (mean, 26.6) enrolled for a drug efficacy trial [38]. Only baseline samples from patients with 205 microscopy- and PCR-confirmed P. falciparum infection were included in this study. 3D7 and Dd2 parasite culture strain mixtures 163 CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 211 In Ecuador, samples (n=41) were collected from clinical patients from March 2019 to April 2020. The 212 samples were collected in Esmeraldas and Carchi Provinces in the north-west of the country, where most 213 P. falciparum cases of Ecuador are reported. All infections were confirmed by microscopy and collected 214 as blood spots in filter paper. Three samples were collected from travelers coming from the Pacific coast 215 in Colombia but diagnosed in Ecuador. 216 217 Data analysis 218 219 For the analysis of the ddPCR data, the following criteria were applied: A minimum of 2 droplets positive 220 for tRNA were required to include a sample in data analysis. Samples were repeated if a deletion was 221 observed but ≤5 droplets were positive for tRNA. At a density of >5 droplets positive for tRNA, the 222 probability of a false-negative result for hrp2 or hrp3 (i.e. no positive droplet in a wild-type infection) is 223 less than 1:500. Data for all samples and assays are provided in Supplementary File S3. 224 225 3D7 and Dd2 parasite culture strain mixtures 163 Even though these 206 samples had been collected nearly a decade ago and might not reflect the current status of hrp2/hrp3 207 deletion, they were included in light of the known high levels of hrp2/hrp3 deletion in Latin America to 208 186 Samples from Zanzibar (n=91) had been collected in the frame of a study on reactive case detection (RCD) 187 from May 2017 to October 2018 [2]. Asymptomatic infections identified through RCD were typed. During 188 RCD, infections were diagnosed by HRP2/pLDH-based RDT (SD BIOLINE Malaria Ag Pf HRP2/pLHD), a blood 189 spot was collected on filter paper, and infections diagnosed by qPCR [2]. Prevalence (not including index 190 cases) was 0.8% by RDT and 2.4% by qPCR. All samples with a density (by qPCR) of >100 parasites/µL were 191 selected for hrp2/hrp3 deletion typing, irrespective of RDT result. 192 186 186 Samples from Zanzibar (n=91) had been collected in the frame of a study on reactive case detection (RCD) 187 from May 2017 to October 2018 [2]. Asymptomatic infections identified through RCD were typed. During 188 RCD, infections were diagnosed by HRP2/pLDH-based RDT (SD BIOLINE Malaria Ag Pf HRP2/pLHD), a blood 189 spot was collected on filter paper, and infections diagnosed by qPCR [2]. Prevalence (not including index 190 cases) was 0.8% by RDT and 2.4% by qPCR. All samples with a density (by qPCR) of >100 parasites/µL were 191 selected for hrp2/hrp3 deletion typing, irrespective of RDT result. 192 ) y y q p y ( y q ) p µ selected for hrp2/hrp3 deletion typing, irrespective of RDT result. 192 193 Samples from Ethiopia (n=47) included clinical cases and asymptomatic individuals sampled in June to 194 November 2016 from Jimma Zone, Oromia Region. P. falciparum prevalence was 4% by microscopy and 195 8.3% by qPCR [36]. No RDT screening was conducted. 196 197 From Ghana, 2 sets of samples were typed. The first set (n=11) was collected in June-September of 2017 198 f f b il h l hild d 5 14 [37] Th d t ( 212) ll t d i M k d 199 Samples from Ethiopia (n=47) included clinical cases and asymptomatic individuals sampled in June to 194 November 2016 from Jimma Zone, Oromia Region. P. falciparum prevalence was 4% by microscopy and 195 8.3% by qPCR [36]. No RDT screening was conducted. 196 . confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Results 226 227 Assay development and validation 228 229 New primers and probes were developed for hrp2 exon 1, hrp2 exon 2, hrp3, and tRNA (Table 1). Upon 230 optimization of assay conditions, clear separation between negative and positive droplets was obtained 231 across a wide range of parasite densities (Figure 1, Supplementary Figure S1). No positive droplets for 232 hrp2 or hrp3 were observed in case of deletion, while the separation between negative droplets and those 233 positive for tRNA remained clear (Figure 1, Supplementary Figure S1). 234 235 236 237 Figure 1: Examples of hrp2 exon 2 deletion typing by ddPCR. Droplets positive for hrp2 are shown in blue (top left of each panel). 238 Droplets positive for tRNA are shown in green (bottom right). Droplets positive for hrp2 and tRNA are shown in orange (top right). 239 Negative droplets (for both hrp2 and tRNA) are shown in gray (bottom left). A) Wild-type infection of medium density. 240 Approximately 600 droplets are positive each for hrp2 and tRNA, and 36 for both targets. B) Wild-type sample of low-density: 32 241 and 29 droplets are positive for hrp2 and tRNA, respectively. C) hrp2 deletion: Droplets are positive for tRNA, but no droplets are 242 A: Wild-type, medium density C: hrp2 deletion B: Wild-type, low density D: Mixed infection negative tRNA positive hrp2 & tRNA positive hrp2 positive 236 A: Wild-type, medium density C: hrp2 deletion B: Wild-type, low density D: Mixed infection negative tRNA positive hrp2 & tRNA positive hrp2 positive A: Wild-type, medium density B: Wild-type, low density B: Wild-type, low density hrp2 & tRNA positive tRNA positive D: Mixed infection C: hrp2 deletion Figure 1: Examples of hrp2 exon 2 deletion typing by ddPCR. Droplets positive for hrp2 are shown in blue (top left of each panel). 238 Droplets positive for tRNA are shown in green (bottom right). . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 Only 15 droplets are positive 243 for hrp2, but 598 droplets are positive for tRNA. 244 To evaluate reproducibility and the limit of reliable detection, 248 isolates from asymptomatic carriers in 246 Kenya were typed for the hrp2 exon 2 assay in triplicate. Geometric mean density was 95 genomes/µL, 247 and 47/248 samples were at densities <10 genomes/µL. By ddPCR, 235/248 (94.6%) of samples met 248 inclusion criteria of ≥2 droplets positive for tRNA in all 3 replicates. Highly similar quantification among 249 replicates was observed (Figure 2A). For each sample, the highest and lowest value of hrp2 copies/µL was 250 recorded. Correlation among technical replicates was very high (R2=0.990). Likewise, for each sample, the 251 highest and lowest value of tRNA copies were recorded, and correlation was very high (R2=0.991). 252 253 254 Figure 2: Validation of assay. A) Samples typed in triplicate for the hrp2 exon 2/tRNA assay. Representative examples of different 255 parasite densities are shown. For each sample, the quantification of hrp2 exon 2, and of tRNA is shown. Results from the same 256 run are connected by a dashed line. B) Mixtures of 3D7 (wild type) and Dd2 (hrp2 deletion). Mixtures were run in triplicate at 257 densities of 10-500 genomes/reaction, and at ratios of 0-100% Dd2. The expected proportion of hrp2 to tRNA copies corresponds 258 to the proportion of 3D7 in the mixture. The observed proportion reflects the expected proportion closely for all mixtures at 500 259 and 100 genome/reaction. confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 Droplets positive for hrp2 and tRNA are shown in orange (top right). 239 Negative droplets (for both hrp2 and tRNA) are shown in gray (bottom left). A) Wild-type infection of medium density. 240 Approximately 600 droplets are positive each for hrp2 and tRNA, and 36 for both targets. B) Wild-type sample of low-density: 32 241 and 29 droplets are positive for hrp2 and tRNA, respectively. C) hrp2 deletion: Droplets are positive for tRNA, but no droplets are 242 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint positive for hrp2. D) Mixed infection with wild-type parasites and parasites carrying hrp2 deletion. Only 15 droplets are positive 243 for hrp2, but 598 droplets are positive for tRNA. 244 245 To evaluate reproducibility and the limit of reliable detection, 248 isolates from asymptomatic carriers in 246 Kenya were typed for the hrp2 exon 2 assay in triplicate. Geometric mean density was 95 genomes/µL, 247 and 47/248 samples were at densities <10 genomes/µL. By ddPCR, 235/248 (94.6%) of samples met 248 inclusion criteria of ≥2 droplets positive for tRNA in all 3 replicates. Highly similar quantification among 249 replicates was observed (Figure 2A). For each sample, the highest and lowest value of hrp2 copies/µL was 250 recorded. Correlation among technical replicates was very high (R2=0.990). Likewise, for each sample, the 251 highest and lowest value of tRNA copies were recorded, and correlation was very high (R2=0.991). 252 positive for hrp2. D) Mixed infection with wild-type parasites and parasites carrying hrp2 deletion. confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 260 Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7 Sample 8 100 101 102 103 104 Genomes/µL hrp2 tRNA 500 genomes 100 genomes 50 genomes 10 genomes 0 20 40 60 80 100 120 140 Genomes per reaction Observed proportion of hrp2 (%) 100% 80% 60% 40% 20% 0% Expected prpoportion of hrp2 (% of 3D7 in mixture) A B 253 254 Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7 Sample 8 100 101 102 103 104 Genomes/µL hrp2 tRNA 500 genomes 100 genomes 50 genomes 10 genomes 0 20 40 60 80 100 120 140 Genomes per reaction Observed proportion of hrp2 (%) 100% 80% 60% 40% 20% 0% Expected prpoportion of hrp2 (% of 3D7 in mixture) A B 500 genomes 100 genomes 50 genomes 10 genomes 0 20 40 60 80 100 120 140 Genomes per reaction Observed proportion of hrp2 (%) 100% 80% 60% 40% 20% 0% Expected prpoportion of hrp2 (% of 3D7 in mixture) B 253 254 Sample 1 Sample 2 Sample 3 Sample 4 Sample 5 Sample 6 Sample 7 Sample 8 100 101 102 103 104 Genomes/µL hrp2 tRNA A B A Genomes per reaction Figure 2: Validation of assay. A) Samples typed in triplicate for the hrp2 exon 2/tRNA assay. Representative examples of different 255 parasite densities are shown. For each sample, the quantification of hrp2 exon 2, and of tRNA is shown. Results from the same 256 run are connected by a dashed line. B) Mixtures of 3D7 (wild type) and Dd2 (hrp2 deletion). Mixtures were run in triplicate at 257 densities of 10-500 genomes/reaction, and at ratios of 0-100% Dd2. The expected proportion of hrp2 to tRNA copies corresponds 258 to the proportion of 3D7 in the mixture. The observed proportion reflects the expected proportion closely for all mixtures at 500 259 and 100 genome/reaction. 260 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint No deletions were detected in the 248 samples from western Kenya. I.e., no samples with droplets for 262 tRNA but no droplets for hrp2 were observed, even though density was low in many samples. In the 263 ddPCR, approximately two-thirds of the reaction volume was transformed into droplets. Applying the 264 threshold of 2 droplets positive for tRNA, three template genomes were required per reaction well to 265 reach that threshold. Up to 9 µL of template DNA could be added to one reaction when primers and 266 probes are kept at 100 µM. Thus, the theoretical limit of detection was 0.33 parasites/µL. If ≤5 droplets 267 are positive for tRNA and a deletion is observed, it is recommended to repeat the sample. 268 269 270 Detection of mixed infections with hrp2-negative and wild type parasites 271 272 To test the ability of the assay to detect deletions when only part of all parasites in an isolate carry the 273 deletion, experimental mixtures were prepared with DNA from parasite culture of 3D7 (wild-type), and 274 Dd2 (hrp2 deletion). Mixtures were prepared at ratios from 0% to 100% Dd2, and at densities of 10-500 275 genomes/reaction. At densities of 100 to 500 genomes/µL, the quantification by ddPCR represented the 276 mixture ratio with high accuracy (Figure 2B). Whenever 40% or more of parasites carried the deletion, the 277 observed ratio was clearly below 100%. In cases where only a small proportion of all parasites carried the 278 deletion (20% Dd2 vs. 80% 3D7), the difference in quantification of hrp2 and tRNA was too small to 279 observe the deletion. Likewise, at densities <50 genomes/reaction the ratio did not accurately reflect the 280 experimental mixture. 281 282 The results were corroborated by screening of 739 field samples from five countries. In wild type isolates, 283 a very similar quantification of hrp2 and tRNA was expected. Unless samples carried a clear deletion (i.e. 284 No deletions were detected in the 248 samples from western Kenya. I.e., no samples with droplets for 262 tRNA but no droplets for hrp2 were observed, even though density was low in many samples. In the 263 ddPCR, approximately two-thirds of the reaction volume was transformed into droplets. confirm the ability of the assays to reliably detect deletions in field isolates [39]. Because of little template 209 volume available, Brazilian samples were not screened for hrp2 exon 1 deletion. 210 Applying the 264 threshold of 2 droplets positive for tRNA, three template genomes were required per reaction well to 265 reach that threshold. Up to 9 µL of template DNA could be added to one reaction when primers and 266 probes are kept at 100 µM. Thus, the theoretical limit of detection was 0.33 parasites/µL. If ≤5 droplets 267 are positive for tRNA and a deletion is observed, it is recommended to repeat the sample. 268 Detection of mixed infections with hrp2-negative and wild type parasites 271 The results were corroborated by screening of 739 field samples from five countries. In wild type isolates, 283 a very similar quantification of hrp2 and tRNA was expected. Unless samples carried a clear deletion (i.e. 284 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint no or very little hrp2 signal), in all samples with densities of >50 copies/µL, the ratio of hrp2 to tRNA copies 285 was above 0.6 (Figure 3). With increasing parasite density, the ratio got closer to 1. 286 287 no or very little hrp2 signal), in all samples with densities of >50 copies/µL, the ratio of hrp2 to tRNA copies 285 was above 0.6 (Figure 3). With increasing parasite density, the ratio got closer to 1. 286 no or very little hrp2 signal), in all samples with densities of >50 copies/µL, the ratio of hrp2 to tRNA c 285 was above 0.6 (Figure 3). With increasing parasite density, the ratio got closer to 1. 286 28 no or very little hrp2 signal), in all samples with densities of >50 copies/µL, the ratio of hrp2 to tRNA copies 285 was above 0.6 (Figure 3). With increasing parasite density, the ratio got closer to 1. 286 287 288 Figure 3: Ratios of hrp2 to tRNA copies in 684 field isolates. With increasing parasite density (X-axis), the ratio becomes close to 289 1. Deletions (with no wild type parasites present) have a ratio of 0. Dashed lines show a ratio of hrp2 to tRNA copies of 0.6, and 290 50 genomes/µL. Mixed infection can be reliably detected at densities >50 genomes/µL, and if >40% of parasites carry the deletion. 291 292 1 10 100 1000 10000 100000 0.0 0.5 1.0 1.5 2.0 2 3 4 5 Genomes/µL Ratio of hrp2 to tRNA copies was above 0.6 (Figure 3). Detection of mixed infections with hrp2-negative and wild type parasites 271 With increasing parasite density, the ratio got closer to 1. 286 was above 0.6 (Figure 3). With increasing parasite density, the ratio got closer to 1. 288 1 10 100 1000 10000 100000 0.0 0.5 1.0 1.5 2.0 2 3 4 5 Genomes/µL Ratio of hrp2 to tRNA copies Figure 3: Ratios of hrp2 to tRNA copies in 684 field isolates. With increasing parasite density (X-axis), the ratio becomes close to 289 1. Deletions (with no wild type parasites present) have a ratio of 0. Dashed lines show a ratio of hrp2 to tRNA copies of 0.6, and 290 50 genomes/µL. Mixed infection can be reliably detected at densities >50 genomes/µL, and if >40% of parasites carry the deletion. 291 292 A lower ratio of hrp2 to tRNA copies was observed in samples from Zanzibar, where the mean ratio of 293 hrp2/tRNA copies was 0.82 in absence of any samples that carried hrp2 deletion (Supplementary Figure 294 S2). The reason is not known, but might be caused by sampling and storage procedures. Blood samples 295 from Zanzibar were collected on filter paper, and stored at ambient temperature for over three years 296 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint prior to extraction. Possibly, this could have resulted in DNA degradation, that affected hrp2 more than 297 tRNA. 298 299 300 Comparison of ddPCR to gel-based nested PCR 301 302 The hrp2 exon 2 ddPCR assay was compared to the classical nPCR assay with visualization of products on 303 agarose gel in 248 asymptomatic infections from western Kenya. The density of asymptomatic infections 304 is often low and thus amplification by PCR can be stochastic. All assays were run in triplicate, i.e. hrp2- 305 exon2/tRNA by ddPCR, hrp2 nested PCR, and msp2 nested PCR. Detection of mixed infections with hrp2-negative and wild type parasites 271 306 307 Samples were included in analysis if the PCR for the control gene was positive in all three replicates, i.e. if 308 >2 droplets were positive for tRNA in the ddPCR assay, or a band was detected for msp2 in all three 309 replicates. For the gel-based assay, 85.5% (212/248) samples met inclusion criteria (Table 2). Among those 310 positive, for 144/212 samples a band for hrp2 was observed in all three replicates. A band in two replicates 311 was observed for 34 samples, and a band in a single replicate for 17 samples. For 17 samples, despite 312 obtaining a band for msp2 in all three replicates, no band for hrp2 was observed. These 17 samples would 313 thus be classified as hrp2 deletion, resulting in a prevalence of deletion of 8.0% (17/212). Figure 3 shows 314 representative examples of results of the nPCR and ddPCR assays. 315 316 Table 2: Comparison between nested PCR and ddPCR-based hrp2 deletion typing 317 318 nested PCR ddPCR Total samples 248 248 prior to extraction. Possibly, this could have resulted in DNA degradati 297 tRNA. 298 299 300 Comparison of ddPCR to gel-based nested PCR 301 302 The hrp2 exon 2 ddPCR assay was compared to the classical nPCR assay 303 agarose gel in 248 asymptomatic infections from western Kenya. The de 304 is often low and thus amplification by PCR can be stochastic. All assay 305 exon2/tRNA by ddPCR, hrp2 nested PCR, and msp2 nested PCR. 306 307 Samples were included in analysis if the PCR for the control gene was po 308 >2 droplets were positive for tRNA in the ddPCR assay, or a band wa 309 replicates. For the gel-based assay, 85.5% (212/248) samples met inclusi 310 positive, for 144/212 samples a band for hrp2 was observed in all three r 311 was observed for 34 samples, and a band in a single replicate for 17 s 312 obtaining a band for msp2 in all three replicates, no band for hrp2 was o 313 thus be classified as hrp2 deletion, resulting in a prevalence of deletion 314 representative examples of results of the nPCR and ddPCR assays. 315 316 Table 2: Comparison between nested PCR and ddPCR-based hrp2 deletion typing 317 318 nested PCR ddPCR Total samples 248 248 prior to extraction. Possibly, this could have resulted in DNA degradation, that affected hrp2 more than 297 tRNA. Detection of mixed infections with hrp2-negative and wild type parasites 271 298 299 300 Comparison of ddPCR to gel-based nested PCR 301 302 The hrp2 exon 2 ddPCR assay was compared to the classical nPCR assay with visualization of products on 303 agarose gel in 248 asymptomatic infections from western Kenya. The density of asymptomatic infections 304 is often low and thus amplification by PCR can be stochastic. All assays were run in triplicate, i.e. hrp2- 305 exon2/tRNA by ddPCR, hrp2 nested PCR, and msp2 nested PCR. 306 The hrp2 exon 2 ddPCR assay was compared to the classical nPCR assay with visualization of products on 303 agarose gel in 248 asymptomatic infections from western Kenya. The density of asymptomatic infections 304 is often low and thus amplification by PCR can be stochastic. All assays were run in triplicate, i.e. hrp2- 305 exon2/tRNA by ddPCR, hrp2 nested PCR, and msp2 nested PCR. 306 307 Samples were included in analysis if the PCR for the control gene was positive in all three replicates, i.e. if 308 >2 droplets were positive for tRNA in the ddPCR assay, or a band was detected for msp2 in all three 309 replicates. For the gel-based assay, 85.5% (212/248) samples met inclusion criteria (Table 2). Among those 310 positive, for 144/212 samples a band for hrp2 was observed in all three replicates. A band in two replicates 311 was observed for 34 samples, and a band in a single replicate for 17 samples. For 17 samples, despite 312 obtaining a band for msp2 in all three replicates, no band for hrp2 was observed. These 17 samples would 313 thus be classified as hrp2 deletion, resulting in a prevalence of deletion of 8.0% (17/212). Figure 3 shows 314 representative examples of results of the nPCR and ddPCR assays. 315 316 Table 2: Comparison between nested PCR and ddPCR-based hrp2 deletion typing 317 318 nested PCR ddPCR Total samples 248 248 Samples were included in analysis if the PCR for the control gene was positive in all three replicates, i.e. if 308 >2 droplets were positive for tRNA in the ddPCR assay, or a band was detected for msp2 in all three 309 replicates. For the gel-based assay, 85.5% (212/248) samples met inclusion criteria (Table 2). Among those 310 positive, for 144/212 samples a band for hrp2 was observed in all three replicates. Detection of mixed infections with hrp2-negative and wild type parasites 271 A band in two replicates 311 was observed for 34 samples, and a band in a single replicate for 17 samples. For 17 samples, despite 312 obtaining a band for msp2 in all three replicates, no band for hrp2 was observed. These 17 samples would 313 thus be classified as hrp2 deletion, resulting in a prevalence of deletion of 8.0% (17/212). Figure 3 shows 314 representative examples of results of the nPCR and ddPCR assays. 315 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Met inclusion criteria 212 (85.5%) 235 (94.8%) Deletion in 3/3 replicates 17 0 Deletion in 2/3 replicates 17 0 Deletion in 1/3 replicates 34 2 No deletion 144 233 Prevalence of deletion 8.0% (17/212) 0% (0/235) Deletion in 3/3 replicates 17 0 Deletion in 2/3 replicates 17 0 Deletion in 1/3 replicates 34 2 No deletion 144 233 Prevalence of deletion 8.0% (17/212) 0% (0/235) 319 320 Figure 3: Comparison of nPCR and ddPCR for hrp2 deletion typing. Representative examples of results obtained by hrp2 and msp2 321 nPCR, and by ddPCR. The expected size of the hrp2 band is 228 bp. A band is visible in samples 3, 4, and 5, but no band is visible 322 in samples 1, 2, 6, and 7. msp2 was run as control for the nPCR assay. msp2 is a size polymorphic gene with amplicons ranging 323 from approximately 200 to 500 bp. Bands are observed for all samples, and multiple bands are observed in case of polyclonal 324 infections. Detection of mixed infections with hrp2-negative and wild type parasites 271 L = 100 bp DNA ladder (New England BioLabs). Samples were run in triplicate, and the same results was obtained all 325 three times. By ddPCR, no deletions were observed in any samples. For samples 1 and 7, the hrp2 exon 2/tRNA ddPCR plot is 326 shown. Droplets are visible for both targets, thus no deletion is observed. 327 hrp2 nPCR msp2 nPCR ddPCR 228 bp (hrp2) L 1 2 3 4 5 6 7 100 200 300 500 100 200 300 500 319 20 hrp2 nPCR msp2 nPCR ddPCR 228 bp (hrp2) L 1 2 3 4 5 6 7 100 200 300 500 100 200 300 500 on of nPCR and ddPCR for hrp2 deletion typing. Representative examples of results obtained by hrp2 and msp2 Figure 3: Comparison of nPCR and ddPCR for hrp2 deletion typing. Representative examples of results obtained by hrp2 and msp2 321 nPCR, and by ddPCR. The expected size of the hrp2 band is 228 bp. A band is visible in samples 3, 4, and 5, but no band is visible 322 in samples 1, 2, 6, and 7. msp2 was run as control for the nPCR assay. msp2 is a size polymorphic gene with amplicons ranging 323 from approximately 200 to 500 bp. Bands are observed for all samples, and multiple bands are observed in case of polyclonal 324 infections. L = 100 bp DNA ladder (New England BioLabs). Samples were run in triplicate, and the same results was obtained all 325 three times. By ddPCR, no deletions were observed in any samples. For samples 1 and 7, the hrp2 exon 2/tRNA ddPCR plot is 326 shown. Droplets are visible for both targets, thus no deletion is observed. 327 Figure 3: Comparison of nPCR and ddPCR for hrp2 deletion typing. Representative examples of resu 321 Figure 3: Comparison of nPCR and ddPCR for hrp2 deletion typing. Representative examples of results obtained by hrp2 and msp2 321 nPCR, and by ddPCR. The expected size of the hrp2 band is 228 bp. A band is visible in samples 3, 4, and 5, but no band is visible 322 in samples 1, 2, 6, and 7. msp2 was run as control for the nPCR assay. msp2 is a size polymorphic gene with amplicons ranging 323 from approximately 200 to 500 bp. Detection of mixed infections with hrp2-negative and wild type parasites 271 Bands are observed for all samples, and multiple bands are observed in case of polyclonal 324 infections. L = 100 bp DNA ladder (New England BioLabs). Samples were run in triplicate, and the same results was obtained all 325 three times. By ddPCR, no deletions were observed in any samples. For samples 1 and 7, the hrp2 exon 2/tRNA ddPCR plot is 326 shown. Droplets are visible for both targets, thus no deletion is observed. 327 329 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint By ddPCR, the criteria for inclusion (≥2 droplets for tRNA) was met by 94.8% (235/248) of samples. Among 330 them, ≥2 droplets for hrp2 were detected in all three replicates in 233/235 samples. In 2 samples in only 331 two of the three replicates ≥2 droplets were positive for hrp2. Both of these samples had one replicate 332 with 1 positive droplet for hrp2, and 5 positive droplets for tRNA. In none of the samples all three or two 333 out of three replicates were negative for hrp2. Thus, the prevalence of deletion by ddPCR was 0%. 334 335 hrp2 and hrp3 deletions in Africa and South America 336 337 The new ddPCR assay was applied to screen for deletions in 830 samples from Kenya, Zanzibar, Ethiopia, 338 Ghana, Brazil, and Ecuador. The frequency of deletions for all loci and sites is given in Table 3. Isolates with deletions of hrp2 and hrp3. Note that these isolates are also counted as deletions in the column Detection of mixed infections with hrp2-negative and wild type parasites 271 339 340 Table 3: hrp2 and hrp3 deletions in Africa and South America 341 Deletions N hrp2 exon 2 hrp2 exon 1 hrp3 hrp2+hrp31 Mixed2 Site Kenya 241 0% (0/241) 0% (0/241) 0% (0/241) 0% (0/241) Zanzibar 91 0% (0/91) 0% (0/91) 0 % (0/91) 0% (0/91) Ethiopia 47 2.1% (1/47) 2.1% (1/47) 74.5% (35/47) 2.1% (1/47) 1 x hrp3 Ghana 223 0% (0/226) 0% (0/223) 0.4% (1/223) 0% (0/170) 1 x hrp2, 3 x hrp3 Brazil 187 46.5% (87/187) NA 62.0% (116/187) 46.0% (86/187) 2 x hrp2, 2 x hrp3 Ecuador 41 0% (0/41) 0% (0/41) 53.7% (22/41) 0% (0/39) 342 1 Isolates with deletions of hrp2 and hrp3. Note that these isolates are also counted as deletions in the columns for hrp2, and for 343 hrp3 (e.g. in Brazil 87 isolates carried hrp2 deletion, of which 86 also carried hrp3 deletion). 344 2 Isolates with only a proportion of all parasites carrying the deletion. 345 Table 3: hrp2 and hrp3 deletions in Africa and South America 341 Deletions N hrp2 exon 2 hrp2 exon 1 hrp3 hrp2+hrp31 Mixed2 Site Kenya 241 0% (0/241) 0% (0/241) 0% (0/241) 0% (0/241) Zanzibar 91 0% (0/91) 0% (0/91) 0 % (0/91) 0% (0/91) Ethiopia 47 2.1% (1/47) 2.1% (1/47) 74.5% (35/47) 2.1% (1/47) 1 x hrp3 Ghana 223 0% (0/226) 0% (0/223) 0.4% (1/223) 0% (0/170) 1 x hrp2, 3 x hrp3 Brazil 187 46.5% (87/187) NA 62.0% (116/187) 46.0% (86/187) 2 x hrp2, 2 x hrp3 Ecuador 41 0% (0/41) 0% (0/41) 53.7% (22/41) 0% (0/39) 342 1 Isolates with deletions of hrp2 and hrp3. Note that these isolates are also counted as deletions in the columns for hrp2, and for 343 hrp3 (e.g. in Brazil 87 isolates carried hrp2 deletion, of which 86 also carried hrp3 deletion). 344 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. Detection of mixed infections with hrp2-negative and wild type parasites 271 ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint From Kenya, 241 samples were typed and no deletions of hrp2 or hrp3 were observed. In Zanzibar, no 347 deletions were observed among 91 samples. Among 223 samples from Ghana, 1 hrp2 deletion/wild type 348 mixed sample was detected (ratio of hrp2 to tRNA copies was <0.6, and parasite density >50 genomes/µL), 349 1 hrp3 deletion, and 3 samples with hrp3 deletion/wild type mixes. 350 351 In Ethiopia, 47 samples met inclusion criteria. One sample carried a deletion of hrp2 exons 1 and 2, 352 resulting in a frequency of deletion of 2.1%. hrp3 deletion was observed in 35/47 (74.5%) samples, and 353 one sample carried a mixed infection with wild type/hrp3 deletion. The sample with hrp2 deletion was 354 among the samples with hrp3 deletion, i.e. both genes were deleted. 355 356 From Brazil, 187 samples were screened. 87 samples carried a deletion of hrp2. Two additional samples 357 carried mixed infections with wild type parasites and hrp2 deletion. Deletion of hrp3 was observed in 358 116/187 (62.0%) samples, and 86/187 (46.0%) samples carried deletions of hrp2 and hrp3. No hrp2 359 deletions were observed in Ecuador, but 22/41 (53.7%) samples carried hrp3 deletions. 360 361 362 From Kenya, 241 samples were typed and no deletions of hrp2 or hrp3 were observed. In Zanzibar, no 347 deletions were observed among 91 samples. Among 223 samples from Ghana, 1 hrp2 deletion/wild type 348 mixed sample was detected (ratio of hrp2 to tRNA copies was <0.6, and parasite density >50 genomes/µL), 349 1 hrp3 deletion, and 3 samples with hrp3 deletion/wild type mixes. 350 In Ethiopia, 47 samples met inclusion criteria. One sample carried a deletion of hrp2 exons 1 and 2, 352 resulting in a frequency of deletion of 2.1%. hrp3 deletion was observed in 35/47 (74.5%) samples, and 353 one sample carried a mixed infection with wild type/hrp3 deletion. The sample with hrp2 deletion was 354 among the samples with hrp3 deletion, i.e. both genes were deleted. 355 From Brazil, 187 samples were screened. 87 samples carried a deletion of hrp2. Two additional samples 57 carried mixed infections with wild type parasites and hrp2 deletion. Deletion of hrp3 was observed in 58 116/187 (62.0%) samples, and 86/187 (46.0%) samples carried deletions of hrp2 and hrp3. No hrp2 59 deletions were observed in Ecuador, but 22/41 (53.7%) samples carried hrp3 deletions. 60 . Detection of mixed infections with hrp2-negative and wild type parasites 271 CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Discussion 363 364 Molecular surveillance of the extent of hrp2 and hrp3 deletions is a high priority task to select the optimal 365 tools for P. falciparum diagnosis. The novel assay based on ddPCR yielded highly accurate results, and was 366 able to detect mixed infections with wild-type parasites and parasites carrying hrp2 deletion. The good 367 performance of the assay was shown by typing of samples from six countries. The samples reflected a 368 range of parasite densities, from low-density asymptomatic infections to high-density clinical infections, 369 and sites represented a range in the frequency of hrp2 and/or hrp3 deletions. Across over 800 field 370 samples screened, <10 samples needed to be repeated because of poor separation between negative and 371 positive droplets, or failure of droplet generation. 372 373 High sensitivity is required for accurate typing of low-density infections. We validated a threshold of two 374 droplets positive for tRNA as limit of detection. When 9 µL of DNA are used as template, this results in a 375 theoretical limit of detection of 0.33 parasites/µL. Sensitivity can be further increased by concentrating 376 the DNA prior to typing. 377 378 The ddPCR assay showed increased sensitivity and specificity to type low-density infections compared to 379 nPCR. Out of 248 asymptomatic samples from western Kenya, 95% could be analyzed by ddPCR, but only 380 85% by nPCR. More importantly, results on deletion status differed substantially. By ddPCR, no deletions 381 were observed. By nPCR, no band was observed for hrp2 in 8% of samples that had a positive band for the 382 control gene (msp2) in all three replicates. Thus, the frequency of deletion was above the 5% threshold, 383 and it would be erroneously recommended to discontinue the use of HRP2-based RDTs. The frequency of 384 deletion by nPCR was similar to a previous study conducted in a nearby site in western Kenya that found 385 hrp2 deletion in 8/89 samples using nPCR [40]. . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint Detection of mixed infections with hrp2-negative and wild type parasites 271 The highly accurate quantification by ddPCR allows 394 detection of mixed infections. Based on experimental mixtures of 3D7 and Dd2, and field samples, mixed 395 infections were reliably detected when at least 40% of parasites carried the deletion, and at densities 396 above 100 genomes per reaction. Using a well-working qPCR assay with an amplification efficacy of 100%, 397 a similar difference between hrp2 and tRNA copy numbers would result in less than half a cycle difference. 398 This is within the normal variation of technical replicates [31, 41], and thus such mixed infections could 399 not be detected by qPCR. 400 401 Twenty-nine isolates from Zanzibar and 16 from Ghana had tested negative by HRP2-based RDT despite 402 high density of 100 to >10,000 parasites/µL [2], yet no hrp2 deletions were observed in these populations. 403 False-negative RDT results might be caused due to incorrect handling, prozone effect [42], or sequence 404 variation without deletion of the hrp2 gene [43]. The finding corroborates the importance of molecular 405 typing. Studies comparing microscopy and RDT results can give important clues for the presence of 406 deletions [44], but molecular typing is required for confirmation [18]. 407 408 No hrp2 deletions were found in Zanzibar, and Kenya, and one mixed infection in Ghana. The data from 409 Ghana contrasts an earlier study that reported a frequency of deletion of >30% [15]. A single hrp2 deletion 410 number of samples points to a possible overestimation of hrp2 deletion frequency by studies relying on 387 nPCR. 388 389 In almost all transmission settings, polyclonal P. falciparum infections are frequent [28]. Using a gel-based 390 assay for deletion typing, in a mixed infection with a wild type parasite and a parasite carrying a deletion, 391 the wild type parasite will produce a band and mask the deletion. These infections can be detected by 392 RDT, but depending on the proportion of polyclonal infections, they can result in pronounced 393 underestimation of the true frequency of deletion [29]. The highly accurate quantification by ddPCR allows 394 detection of mixed infections. Based on experimental mixtures of 3D7 and Dd2, and field samples, mixed 395 infections were reliably detected when at least 40% of parasites carried the deletion, and at densities 396 above 100 genomes per reaction. Detection of mixed infections with hrp2-negative and wild type parasites 271 The direct comparison of ddPCR and nPCR in a large 386 Discussion 363 364 Molecular surveillance of the extent of hrp2 and hrp3 deletions is a high priority task to select the optimal 365 tools for P. falciparum diagnosis. The novel assay based on ddPCR yielded highly accurate results, and was 366 able to detect mixed infections with wild-type parasites and parasites carrying hrp2 deletion. The good 367 performance of the assay was shown by typing of samples from six countries. The samples reflected a 368 range of parasite densities, from low-density asymptomatic infections to high-density clinical infections, 369 and sites represented a range in the frequency of hrp2 and/or hrp3 deletions. Across over 800 field 370 samples screened, <10 samples needed to be repeated because of poor separation between negative and 371 positive droplets, or failure of droplet generation. 372 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint number of samples points to a possible overestimation of hrp2 deletion frequency by studies relying on 387 nPCR. 388 389 In almost all transmission settings, polyclonal P. falciparum infections are frequent [28]. Using a gel-based 390 assay for deletion typing, in a mixed infection with a wild type parasite and a parasite carrying a deletion, 391 the wild type parasite will produce a band and mask the deletion. These infections can be detected by 392 RDT, but depending on the proportion of polyclonal infections, they can result in pronounced 393 underestimation of the true frequency of deletion [29]. Detection of mixed infections with hrp2-negative and wild type parasites 271 Using a well-working qPCR assay with an amplification efficacy of 100%, 397 a similar difference between hrp2 and tRNA copy numbers would result in less than half a cycle difference. 398 This is within the normal variation of technical replicates [31, 41], and thus such mixed infections could 399 not be detected by qPCR. 400 No hrp2 deletions were found in Zanzibar, and Kenya, and one mixed infection in Ghana. The data from 409 Ghana contrasts an earlier study that reported a frequency of deletion of >30% [15]. A single hrp2 deletion 410 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. ch was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: xiv preprint was found in southwestern Ethiopia. This is in stark contrast to very high levels of deletion in western 411 Ethiopia [45], Eritrea [18], and Sudan [46]. The results corroborate the need for studies assessing hrp2 412 deletion and selection of diagnostic tools at sub-national level. 413 was found in southwestern Ethiopia. This is in stark contrast to very high levels of deletion in western 411 Ethiopia [45], Eritrea [18], and Sudan [46]. The results corroborate the need for studies assessing hrp2 412 deletion and selection of diagnostic tools at sub-national level. 413 414 Contrasting findings were obtained from the sample sets from South America, with very high levels of 415 deletion in Brazil, and no deletions in Ecuador. Brazil and Ecuador share no borders, and the amount of 416 human migration is limited. The results add to the heterogenous pattern of hrp2/hrp3 deletion in South 417 America, with high frequency of hrp2 deletion in the Amazon [9, 12, 39], but low levels among the Pacific 418 coast [11, 12]. An outbreak of parasites with hrp2 deletion at the Peruvian Pacific coast was caused by 419 infections imported from the Amazon [47]. Brazil is committed to P. Detection of mixed infections with hrp2-negative and wild type parasites 271 falciparum elimination, and the 420 samples typed originated from the main hotspot of transmission [48]. Rapid diagnostic tests (RDTs) remain 421 relatively little used in Brazil. Microscopy remains the diagnostic method of choice, and RDTs are mostly 422 used in remote areas, e.g., populations from Amerindian Reserves and some traditional riverine 423 communities with no access to conventional microscopy. The frequency of hrp2 deletion in Brazil clearly 424 exceeds the 5% threshold, thus it is recommended that no HRP2-based RDTs are used. 425 426 Ethiopia [45], Eritrea [18], and Sudan [46]. The results corroborate the need for studies assessing hrp2 412 deletion and selection of diagnostic tools at sub-national level. 413 414 Contrasting findings were obtained from the sample sets from South America, with very high levels of 415 deletion in Brazil, and no deletions in Ecuador. Brazil and Ecuador share no borders, and the amount of 416 human migration is limited. The results add to the heterogenous pattern of hrp2/hrp3 deletion in South 417 America, with high frequency of hrp2 deletion in the Amazon [9, 12, 39], but low levels among the Pacific 418 coast [11, 12]. An outbreak of parasites with hrp2 deletion at the Peruvian Pacific coast was caused by 419 infections imported from the Amazon [47]. Brazil is committed to P. falciparum elimination, and the 420 samples typed originated from the main hotspot of transmission [48]. Rapid diagnostic tests (RDTs) remain 421 relatively little used in Brazil. Microscopy remains the diagnostic method of choice, and RDTs are mostly 422 used in remote areas, e.g., populations from Amerindian Reserves and some traditional riverine 423 communities with no access to conventional microscopy. The frequency of hrp2 deletion in Brazil clearly 424 exceeds the 5% threshold, thus it is recommended that no HRP2-based RDTs are used. 425 426 To determine whether the frequency of the deletion exceeds the threshold of 5%, the WHO recommends 427 that for each site a minimum of 370 is typed [49]. Limitations of the molecular assays available for typing 428 have been a major hindrance to type that number of samples in many sites where hrp2 or hrp3 deletion 429 is suspected. Even a low proportion of false-negative results could impact the decision to discontinue 430 HRP2-based RDTs. Detection of mixed infections with hrp2-negative and wild type parasites 271 As a result of the scarcity of field data, the spatiotemporal dynamics of hrp2 deletion 431 in parasite populations, drivers of the deletion, potential fitness costs, and clinical consequences remain 432 poorly understood. Results from simulation studies suggest that the use of hrp2-based RDTs selects for 433 Contrasting findings were obtained from the sample sets from South America, with very high levels of 415 deletion in Brazil, and no deletions in Ecuador. Brazil and Ecuador share no borders, and the amount of 416 human migration is limited. The results add to the heterogenous pattern of hrp2/hrp3 deletion in South 417 America, with high frequency of hrp2 deletion in the Amazon [9, 12, 39], but low levels among the Pacific 418 coast [11, 12]. An outbreak of parasites with hrp2 deletion at the Peruvian Pacific coast was caused by 419 infections imported from the Amazon [47]. Brazil is committed to P. falciparum elimination, and the 420 samples typed originated from the main hotspot of transmission [48]. Rapid diagnostic tests (RDTs) remain 421 relatively little used in Brazil. Microscopy remains the diagnostic method of choice, and RDTs are mostly 422 used in remote areas, e.g., populations from Amerindian Reserves and some traditional riverine 423 communities with no access to conventional microscopy. The frequency of hrp2 deletion in Brazil clearly 424 exceeds the 5% threshold, thus it is recommended that no HRP2-based RDTs are used. 425 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . Detection of mixed infections with hrp2-negative and wild type parasites 271 CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint hrp2-negative parasites, in particular if transmission levels are low and a large proportion of all infections 434 turn clinical and result in treatment seeking [50, 51]. 435 436 In conclusion, the novel, high-throughput, highly sensitive and specific ddPCR assay will facilitate 437 molecular surveillance for hrp2 and hrp3 deletion, and thus aid selection of diagnostic tests to accelerate 438 malaria control and elimination. Data obtained using this assay will help to understand the evolutionary 439 processes underlying the de-novo emergence and spread of the deletion. 440 441 442 Acknowledgements 443 We thank all study participants providing blood samples and the study teams and health center personnel 444 who supported sample collection. We thank Michael T. Ferdig and Katelyn M. Vendrely for providing 445 culture strain DNA for the 3D7/Dd2 experimental mixtures. 446 447 448 Supplementary Files 449 Supplementary File S1: Assay conditions 450 Supplementary File S2: Supplementary Figures S1, S2 451 Supplementary File S3: Database 452 453 454 Financial Disclosure Statement 455 hrp2-negative parasites, in particular if transmission levels are low and a large proportion of all infections 434 turn clinical and result in treatment seeking [50, 51]. 435 hrp2-negative parasites, in particular if transmission levels are low and a large proportion of all infections 434 turn clinical and result in treatment seeking [50, 51]. 435 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. Detection of mixed infections with hrp2-negative and wild type parasites 271 not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: nt This work was supported by NIH grants R21AI137891 awarded to CK, and U19 AI129326, D43 TW001505 456 awarded to GY (https://www.nih.gov). The funders had no role in study design, data collection and 457 analysis, decision to publish, or preparation of the manuscript. 458 459 460 Competing interests 461 The authors state that no competing interests exist. 462 463 This work was supported by NIH grants R21AI137891 awarded to CK, and U19 AI129326, D43 TW001505 456 awarded to GY (https://www.nih.gov). The funders had no role in study design, data collection and 457 analysis, decision to publish, or preparation of the manuscript. 458 . CC-BY 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. (which was not certified by peer review) The copyright holder for this preprint this version posted June 4, 2021. ; https://doi.org/10.1101/2021.06.01.21258117 doi: medRxiv preprint References 464 465 1. World Health Organisation. World Malaria Report 2020. 2020. 466 2. 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https://openalex.org/W4309214759	https://www.frontiersin.org/articles/10.3389/fpls.2022.1035620/pdf	English	null	QTL analysis identified two major all-internodes solidness loci from a completely solid-stemmed spring wheat line	Frontiers in plant science	2,022	cc-by	16,893	OPEN ACCESS OPEN ACCESS EDITED BY Jose Miguel Soriano, Institute of Agrifood Research and Technology (IRTA), Spain REVIEWED BY Aduragbemi Amo, University of Georgia, United States Peter Bulli, Jaramogi Oginga Odinga University of Science and Technology, Kenya Ljiljana Kuzmanovic, University of Tuscia, Italy CORRESPONDENCE Harpinder S. Randhawa harpinder.randhawa@agr.gc.ca SPECIALTY SECTION This article was submitted to Plant Breeding, a section of the journal Frontiers in Plant Science RECEIVED 03 September 2022 ACCEPTED 27 September 2022 PUBLISHED 15 November 2022 EDITED BY Jose Miguel Soriano, Institute of Agrifood Research and Technology (IRTA), Spain Raman Dhariwal 1, Colin W. Hiebert 2 and Harpinder S. Randhawa 1 1Agriculture and Agri-Food Canada, Lethbridge Research and Development Centre, Lethbridge, AB, Canada, 2Agriculture and Agri-Food Canada, Morden Research and Development Centre, Morden, MB, Canada The culms of solid-stemmed wheat cultivars are ﬁlled with “pith” – a parenchymatous tissue largely composed of soft, spongy, and compact parenchyma cells. Breeding solid-stemmed cultivars is the most effective way to decrease the detrimental impact of wheat stem sawﬂy (WSS), Cephus cinctus Norton (Hymenoptera: Cephidae) on wheat production. Although a major solid stem gene has been previously identiﬁed from durum wheat, it produces an intermediate level of stem solidness in common wheat which is insufﬁcient to provide the required level of WSS resistance. The maximum resistance is achieved when stems are totally ﬁlled with pith. Thus, to identify a secondary source of solidness in common wheat, we developed three mapping populations from wheat cvs. Sadash, ‘AAC Innova’ and ‘AAC Cameron’, each crossed separately with P2711, a completely solid-stemmed hexaploid wheat breeding line. All populations were genotyped using either wheat 15K or 90K Inﬁnium iSelect SNP Assay and high-density linkage maps were generated from individual populations along with consensus maps for chromosomes 3B and 3D from all populations. ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations were subjected to extensive phenotyping in ≥3 environments followed by quantitative trait loci (QTL) analyses using population-speciﬁc and consensus linkage maps. We identiﬁed two major solid stem QTLs in the distal regions of chromosome arms 3BL and 3DL in both populations in addition to several population-speciﬁc or common minor QTLs. Internode-speciﬁc QTL analyses detected both major QTLs of chromosomes 3B and 3D across internodes, from top to bottom of the stalk, but minor QTLs were largely detected in upper or middle internodes. TYPE Original Research PUBLISHED 15 November 2022 DOI 10.3389/fpls.2022.1035620 TYPE Original Research PUBLISHED 15 November 2022 DOI 10.3389/fpls.2022.1035620 QTL analysis identiﬁed two major all-internodes solidness loci from a completely solid- stemmed spring wheat line OPEN ACCESS EDITED BY Jose Miguel Soriano, Institute of Agrifood Research and Technology (IRTA), Spain REVIEWED BY Aduragbemi Amo, University of Georgia, United States Peter Bulli, Jaramogi Oginga Odinga University of Science and Technology, Kenya Ljiljana Kuzmanovic, University of Tuscia, Italy CORRESPONDENCE Harpinder S. Randhawa harpinder.randhawa@agr.gc.ca SPECIALTY SECTION This article was submitted to Plant Breeding, a section of the journal Frontiers in Plant Science RECEIVED 03 September 2022 ACCEPTED 27 September 2022 PUBLISHED 15 November 2022 CITATION Dhariwal R, Hiebert CW and Randhawa HS (2022) QTL analysis identiﬁed two major all-internodes solidness loci from a completely solid- stemmed spring wheat line. F Pl S i 13 1035620 TYPE Original Research PUBLISHED 15 November 2022 DOI 10.3389/fpls.2022.1035620 Introduction 1926; Varella et al., 2017). These wheat cultivars are referred to as solid stemmed wheat and exhibit resistance by deterring oviposition for WSS adult females, impeding larval migration and growth or death of larvae inside stems, which greatly reduce stem cutting and WSS population abundance (Wallace et al., 1973). The use of solid-stemmed cultivars is considered the most effective way to minimize WSS damage (Beres et al., 2013). Wheat stem sawﬂy (WSS), Cephus cinctus Norton (Hymenoptera: Cephidae), imposes severe economic barriers to wheat (Triticum aestivum L.) production in the northern great plains of North America and many other wheat-producing regions around the globe (Morrill et al., 1992; Talbert et al., 2014). WSS females deposit eggs in hollow stems soon after emergence from the infested stubble of the previous year’s crop following mating, usually in the early crop growing season (Beres et al., 2011a; Talbert et al., 2014; Subedi et al., 2020). These eggs hatch within 5–7 d of deposition and the resulting larvae begins feeding upon the vascular bundle tissues within the stalk (Ainslie, 1929). Larvae feed on the vascular bundles, make tunnel across internodes in the stem, interfere with the stem storage and transfer of water and nutrients to the developing grains, and reduce photosynthesis and grain yield (Macedo et al., 2007). In recent years, WSS has not only expanded its infestation area outside its traditional territories (Lesieur et al., 2016) but has also become a constant economically challenging insect pest in North America (Subedi et al., 2020). WSS-infested plants showed 5–30% kernel weight reductions in ﬁeld and greenhouse conditions (Wallace et al., 1973; Delaney et al., 2010). In high WSS pressure, up to ~15% additional yield loss can be observed when infested/cut stems are lodged on the ground or healthy stems in ﬁeld conditions, reaching a total loss of around 30–35% (Beres et al., 2007; Bekkerman and Weaver, 2018). Harvest losses attributed to WSS are estimated to exceed $350 million annually in North America alone (Beres et al., 2011b). Research on stem solidness has been the major focus of WSS resistance in breeding programs for over several decades. OPEN ACCESS Our results suggest that both major QTLs are sufﬁcient to develop highly solid- stemmed cvs; however, the minor loci, which additively enhance the pith expression, can be coupled with major genes to achieve a complete solid stem phenotype in common wheat. Comparative and haplotype analyses showed ACCEPTED 27 September 2022 PUBLISHED 15 November 2022 CITATION Dhariwal R, Hiebert CW and Randhawa HS (2022) QTL analysis identiﬁed two major all-internodes solidness loci from a completely solid- stemmed spring wheat line. Front. Plant Sci. 13:1035620. doi: 10.3389/fpls.2022.1035620 COPYRIGHT © 2022, His Majesty the King in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada for the contribution of Dhariwal, C. Hiebert, and H.S. Randhawa. This is an open- access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Frontiers in Plant Science 01 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 10.3389/fpls.2022.1035620 that the 3B locus is homoeologous to 3D, the former being mapped to a 1.1 Mb genomic region. Major QTLs identiﬁed in this study can be incorporated in modern wheat cultivars to achieve maximum WSS resistance from high pith expression. wheat stem sawﬂy, solid stem wheat, pith development, SNP mapping, linkage map, QTL mapping Frontiers in Plant Science Dhariwal et al. Introduction 10.3389/fpls.2022.1035620 impacted areas in the late 1940s and early 1950s (Talbert et al., 2014). In the following decades, superior solid-stemmed cultivars, particularly durum wheat and some common wheat, quickly followed and dominated acreage in WSS-infested regions in North America (Talbert et al., 2014). identiﬁcation of additional gene(s) that increase the stem- solidness in P2711 and their linked markers would be useful for common wheat breeding. For this study, we developed two doubled haploid (DH) populations and one recombinant inbred line (RIL) population from Canadian common wheat cvs. Sadash (Sadasivaiah et al., 2009), ‘AAC Innova’ (Randhawa et al., 2015) and ‘AAC Cameron’ (Fox et al., 2016), each crossed with P2711 (Farzand et al., 2022). The objectives of this study were to identify secondary QTLs and their markers for stem-solidness from P2711 that would enhance the effect of the primary QTL and compare identiﬁed QTLs with the ones reported previously. Genetic experiments started in the early 21st century in hexaploid wheat have identiﬁed a major stem solidness QTL Qss.msub-3BL (Cook et al., 2004) and a moderate effect locus Qss.msub-3DL (Lanning et al., 2006). The 3BL major locus has also been detected in durum wheat (T. turgidum var. durum), which was later designated as Solid-stem locus 1 (SSt1) (Houshmand et al., 2007). Recently, Cook et al. (2017) revealed several new haplotypes at Qss.msub.3BL such as one for the early stem solidness. In addition to 3BL and 3DL loci, a number of minor QTLs that affects solid stem expression have been detected in common and durum wheat (Larson and Macdonald, 1959a; Sherman et al., 2010; Varella et al., 2015; Nilsen et al., 2017; Varella et al., 2019). Kebrom et al. (2012) discovered that a ‘tiller inhibition’ (tin) gene (Atsmon and Jacobs, 1977) that is tightly linked with the simple sequence repeat (SSR) marker gwm136 on chromosome arm 1AS (Spielmeyer and Richards, 2004) is associated with precocious internode development and solidness in basal internodes of wheat plants. Later in a separate study, Hyles et al. (2017) found that tin encodes a cellulose synthase-like (Csl) protein which increases ligniﬁcation associated with stronger stems in tin wheat plants. In 2017, Oiestad et al., (2017) reported that stem solidness at Qss.msub-3BL locus is associated with gene expression changes related to lignin biosynthesis. Recently, Nilsen et al. Plant material Three spring wheat mapping populations, two DH and one RIL, were developed by crossing Canadian common wheat cvs. Sadash, ‘AAC Innova’ and ‘AAC Cameron’, each used as female with P2711 as a male parent. F1s produced from ‘Sadash/P2711’ and ‘AAC Innova/P2711’ crosses were used for doubled haploid production using the wheat-maize pollination technique (Suenaga and Nakajima, 1989) at the Agriculture and Agri- Food Canada (AAFC), Lethbridge Research and Development Centre (LeRDC), Lethbridge, AB, Canada. F1s produced from the ‘AAC Cameron/P2711’ cross were used to produce the RIL population as described earlier (Farzand et al., 2022). The numbers of recombinant lines produced from these three crosses were 180 (‘Sadash/P2711’), 374 (‘AAC Innova/P2711’), and 252 (‘AAC Cameron/P2711’). Sadash is a hollow-stemmed, white-grained, semi-dwarf, high-yielding, soft white spring wheat cultivar which belongs to the ‘Canada Western Soft White Spring’ (CWSWS) market class. It was developed using the traditional plant breeding method from the cross ‘SWS207/SWS208//SWS214’ made at the AAFC-LeRDC in 1997. ‘AAC Innova’ is a hollow-stemmed, white-grained, semi-dwarf, soft white spring type cultivar which belongs to the ‘Canada Western Special Purpose’ (CWSP) market class. ‘AAC Innova’ originated from the cross ‘AC Andrew/N9195’ made at AAFC-LeRDC in 2001 and was developed using a modiﬁed bulk breeding technique (Randhawa et al., 2015). ‘AAC Cameron’ is a hollow-stemmed, red-grained, tall and high-yielding spring wheat cultivar with good agronomic, disease, and end-use quality characteristics which belongs to the ‘Canada Western Red Spring’ (CWRS) market class (Fox et al., 2016). ‘AAC Cameron’ was developed using the modiﬁed pedigree breeding method from the complex cross ‘D1125/Alsen//BW346/3/BW370/99B60-EJ26’ made at AAFC, Cereal Research Centre (CRC), Winnipeg, Manitoba (MB) in 2004 (Fox et al., 2016). Male parent P2711 is a completely solid-stemmed (for all internodes), semi-dwarf, Despite the progress made in the research, breeding hexaploid wheat for stem-solidness is a challenge due to inconsistent expression of pith in hexaploid genetic backgrounds (Cook et al., 2004; Lanning et al., 2006). Stems of genotypes containing the allele for solidness often are of an intermediate solidness in common wheat that does not confer sufﬁcient resistance to the WSS (Wallace et al., 1973; Morrill et al., 1992; Lanning et al., 2006). This is perhaps because most solid‐stemmed common wheat genotypes carry a single solid stem gene from the Portuguese landrace ‘S‐615’ (Varella et al., 2019) which does not fully express in the hexaploid background due to some inhibitors and epistatic interactions. Introduction The ﬁrst genetic study on solid stem inheritance in wheat was published in 1905 by Biffen (1905) followed by a series of classical genetics studies on different species and cultivars in succeeding years (Engledow, 1920; Engledow, 1923; Engledow and Hutchinson, 1925; Thompson, 1931; Kemp, 1934; Goytia Y Angulo, 1935; Yamashita, 1937; Platt, 1941; Putnam, 1942; Platt and Larson, 1944; Larson, 1952; Mc Neal, 1956; Mcneal et al., 1957; Larson, 1959a; Larson, 1959b; Larson, 1959c; Larson and Macdonald, 1959a; Larson and Macdonald, 1959b; Mc Neal, 1961; Bozzini and Avanzi, 1962; Larson and Macdonald, 1962; Mckenzie, 1965). These studies not only helped in the identiﬁcation of genomes and chromosomes carrying genetic factors/genes and their number variation that determines stem solidness or hollowness but also provided a solid background work on resistance to the WSS due to solid stem. These studies also demonstrated that stem solidness was most common in the tetraploid species, while unknown in diploids (Vavilov, 1951) and rare in hexaploids (Platt et al., 1941). Moreover, durum wheat was found more solid than common wheat (Kemp, 1934). This steered intensive efforts for introgression or transfer of the complete stem solidness from tetraploid to hexaploid species as a resistance source to control the WSS, which resulted in large failures initially (Platt and Larson, 1944; Larson, 1959c) but succeeded later on in the speciﬁc genetic background (Mc Neal, 1961). Instantaneously, wheat breeding for resistance to the WSS had become synonymous with breeding for stem solidness (Talbert et al., 2014). The ﬁrst North American hexaploid (hard red spring) wheat cultivar released from these efforts in Canada was ‘Rescue’, which was quickly picked up by the growers and grown throughout the wheat stem sawﬂy Due to the speciﬁc lifecycle and presence of alternate hosts, WSS control measures are limited (Weiss and Morrill, 1992; Beres et al., 2011a; Talbert et al., 2014). Adult females emerge for a short period, and WSS larvae feed and develop inside the stem where they are protected from the external environment including insecticide applications, thus pest management strategies primarily rely on host plant resistance (Talbert et al., 2014; Biyiklioglu et al., 2018). Initial research on Cephus pygmaeus (L.), a species of the European WSS, in the 1920s revealed less damage in wheat plants having stems ﬁlled with pith (Schegolev, Frontiers in Plant Science 02 frontiersin.org Dhariwal et al. Introduction (2020) identiﬁed TdDof, which encodes Dof zinc ﬁnger protein, as the causal gene that controls stem solidness at the SSt1 locus on chromosome 3B in durum. They showed that copy number gain of TdDof correlates with its increased expression and the solid-stem phenotype. Frontiers in Plant Science Phenotypic data analysis The analysis of variance (ANOVA) of solid stem for each environment and pooled data was assessed using the line solid stem score as demonstrated earlier by Basnet et al. (2012) and (Farzand et al., 2022). Brieﬂy, ANOVA was conducted by treating genotypes within environments as random effect factors, while experiments as ﬁxed effect factors using the packages ‘lme4’ (version 1.1.27.1) (Bates et al., 2015) and ‘lattice’ (version 0.20-45) (Sarkar, 2008) in R (version 4.0.3) (R Core Team, 2013). Covariance parameter estimates as unbiased genetic variance component estimates were used for calculating broad sense heritability (H2) as the ratio of genetic variance to phenotypic variance following Knapp et al. (1985). Pearson correlation plots, histograms with normal curves and scatterplots were estimated and plotted using the R packages GGally (Schloerke et al., 2018), randomForest (Liaw and Wiener, 2002) and ggplot2 (Wickham, 2016) in R (version 4.0.3) (R Core Team, 2013). Least-square means (LS means) for stem-solidness score (internodes and line) were estimated for each recombinant doubled haploid line using Microsoft’s Excel program. Plant material Five tillers, separately from each of the three to ﬁve plants (at least 5 x 3 = 15 total tillers or stems), were dissected longitudinally and four upper internodes (in top-down order: I1: internode 1/peduncle, I2: internode 2, I3: internode 3, and I4: internode 4/lowest internode scored) were rated for a stem- solidness (S) score of 1–5 (1 for hollow stem – no pith development, 2 for some degree of pith development, 3 for the large hollow tunnel in the stem or a huge cavity at a particular point, 4 for the size of hollow tunnel equivalent to a pencil lead, and 5 for completely solid stem). Scores of each internode across all tillers/stems were averaged to obtain a rating score per internode. While stem solidness score from each internode was utilized separately for QTL analysis to identify internode- speciﬁc loci, a plot/line score obtained by averaging all four internode scores was utilized for combined QTL analysis for stem solidness. Similarly, the rating was done for greenhouse- grown plants but the number of plants and tillers/stems per line varied. At least one plant and 5 tillers or stems were rated in the greenhouse. Common wheat cv Lillian (Depauw et al., 2005) and durum cv ‘Golden Ball’ (Clark et al., 1922) were used as checks for comparisons. Lillian is a hard red spring semi-solid-stemmed wheat cultivar which exhibits reduced WSS cutting (Depauw et al., 2005), while Golden Ball is a completely solid-stemmed durum cultivar (Kemp, 1934). Seeds of cultivars used as checks and parents of the mapping populations were accessed from the Wheat Breeding core collection at AAFC-LeRDC. Seeds of parents, DHs and RILs produced in this study are preserved at AAFC-LeRDC and available upon request. All other cultivars used in this study are preserved at the Plant Gene Resources of Canada (PGRC) seed genebank based at AAFC’s Saskatoon Research and Development Centre, Saskatchewan, Canada. Plant material Maximum resistance from stem solidness can be obtained when the stems are completely ﬁlled with pith (Holmes, 1977). Thus, to develop complete solid-stemmed cultivars of hexaploid wheat, it is necessary to identify the secondary sources of stem solidness. P2711 is a completely solid-stemmed (Supplementary Video S1) hexaploid spring wheat breeding line from South Africa but has not yet been characterized for stem solidness. Thus, the Frontiers in Plant Science 03 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 10.3389/fpls.2022.1035620 1982) and a modiﬁed phenotyping method (Dhariwal et al., 2017). Brieﬂy, three to ﬁve plants from the center of each plot/ row were pulled from the ﬁeld at maturity and moved to the greenhouse where they were left on open benches for a few weeks for complete drying before rating for stem-solidness/pith expression. Five tillers, separately from each of the three to ﬁve plants (at least 5 x 3 = 15 total tillers or stems), were dissected longitudinally and four upper internodes (in top-down order: I1: internode 1/peduncle, I2: internode 2, I3: internode 3, and I4: internode 4/lowest internode scored) were rated for a stem- solidness (S) score of 1–5 (1 for hollow stem – no pith development, 2 for some degree of pith development, 3 for the large hollow tunnel in the stem or a huge cavity at a particular point, 4 for the size of hollow tunnel equivalent to a pencil lead, and 5 for completely solid stem). Scores of each internode across all tillers/stems were averaged to obtain a rating score per internode. While stem solidness score from each internode was utilized separately for QTL analysis to identify internode- speciﬁc loci, a plot/line score obtained by averaging all four internode scores was utilized for combined QTL analysis for stem solidness. Similarly, the rating was done for greenhouse- grown plants but the number of plants and tillers/stems per line varied. At least one plant and 5 tillers or stems were rated in the greenhouse. red-grained hexaploid spring wheat breeding line from South Africa but has not yet been characterized for stem solidness. 1982) and a modiﬁed phenotyping method (Dhariwal et al., 2017). Brieﬂy, three to ﬁve plants from the center of each plot/ row were pulled from the ﬁeld at maturity and moved to the greenhouse where they were left on open benches for a few weeks for complete drying before rating for stem-solidness/pith expression. Trial environments and ﬁeld experiments ‘Sadash/P2711’recombinantdoubledhaploidlines,theirparents and check cultivars were grown in three environments at AAFC- LeRDC, Lethbridge (49°41′N, 112°49′W) in Alberta, Canada. The threeenvironmentsincluded:(i)ﬁeldtrialin2014(LTFD14),(ii)ﬁeld trial in 2019 (LTFD19), and (iii) greenhouse trial in 2019 (LTGH19). ‘AAC Innova/P2711’ recombinant doubled haploid lines, their parents and check cultivars were grown in one and three environments, respectively, in Lincoln (43°63′S, 172°46′E), New Zealand and AAFC-LeRDC, Lethbridge, AB. The four environments included: (i) Lincoln, New Zealand ﬁeld trial in 2013 (NZFD13), (ii) Lethbridge ﬁeld trial in 2015 (LTFD15), (iii) Lethbridge greenhouse trial in 2015 (LTGH15), and (iv) Lethbridge ﬁeld trial in 2019 (LTFD19). ‘AAC Cameron/P2711’ RILs, their parents and check cultivars were grown in the ﬁeld in one and two environments, respectively, in Lincoln, New Zealand and AAFC-LeRDC, Lethbridge, AB. The three environments included: (i) F4 ﬁeld trial in Lethbridge in 2015 (LTFD15), (ii) F5 ﬁeld trial in Lincoln, New Zealand in 2016 (NZFD16), and (iii) F6 ﬁeld trial at AAFC-Lethbridge in 2017 (LTFD17). Field trials in both locations were conducted on conventionally managed ﬁelds, while greenhouse experiments were conducted using a 16/8h light/dark photoperiod and the day/night temperature of 23/18°C. Single 3-m-long rows of each line/genotype were seeded in late May in a randomized block design in all Lethbridge environments. Seeding density was ~250 seeds per row with 23.5 cm row spacing. Hill plots were seeded in mid-October in New Zealand, while two pots were seeded, each with two plants, for each line/genotype in greenhouse experiments. In New Zealand ﬁeld trials, solid-stem rating was done in the ﬁeld by cutting erect wheat plants/stems at several points using sharp scissors. A subjective rating was given to lines on visual bases. Using this method, the best solid-stemmed line was rated with a solid stem score of 3 while the worst with a solid stem score of 1. Frontiers in Plant Science Genotyping, linkage mapping and quantitative trait loci analysis For isolation of DNA from leaf samples, four seeds were sown from each genotype in 96 cell seed planting trays (parents and checks repeated over all trays) in a soil mixture of Turface (9.07 kg), Peat Moss (0.907 kg) and Vermiculite (0.06 m3). Leaf tissue samples were collected from 10 days old seedlings followed by isolation of DNA using DNeasy 96 Plant Kit (Qiagen Inc., Valencia, CA, USA) following Dhariwal et al. (2018). Quant-iT™PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientiﬁc Inc., Bartlesville, OK, USA) was used to quantify DNA samples followed by dilutions of DNA samples to 50 ng/µl. ‘Sadash/P2711’ population, its parents and checks were genotyped using Wheat 15K Illumina Inﬁnium SNP array from TraitGenetics GmbH, Germany. ‘AAC Innova/P2711’ and ‘AAC Cameron/P2711’ populations, their parents and checks were genotyped using a 90K Illumina Inﬁnium array (Wang et al., 2014). Single nucleotide polymorphism (SNP) genotyping data were analyzed using Genotyping module of GenomeStudio software (Illumina Inc., CA, USA) as described previously (Dhariwal et al., 2020). Brieﬂy, high quality SNPs were selected from the list of all SNPs evaluated for genotyping using following ﬁlters in GenomeStudio (i) Call Freq: >0.50, (ii) Minor Allele Freq: >0.03, (iii) AA Freq:!=1, (iv) AB Freq:!=1, and (v) BB Freq:!=1. Monomorphic and SNPs that deviated signiﬁcantly from the 1:1 ratio were removed by comparing the parental proﬁle and using the c2 test, respectively. Composite interval mapping (CIM) to identify the main effect solid stem QTLs (for the combined effect of all internodes or whole stem as well as internode speciﬁc analyses) in all individual environments and pooled (across environments) data collected for each population were conducted separately following Dhariwal and Randhawa (2022). The regression method “forwards and backwards cofactor (p < 0.05)” was used for CIM in the software package QTL Cartographer (version 1.6) (Zeng, 1993; Zeng, 1994) following Nilsen et al. (2017). Signiﬁcant QTLs were declared using QTL thresholds estimated using 1000 permutations at a signiﬁcance level of p = 0.05. QTL conﬁdence intervals were determined using 1-LOD and 2-LOD support intervals as 95% and 99% CI (Lander and Botstein, 1989). Linkage bins possessing two or more QTLs within 10.0 cM were treated as a single QTL region (Dhariwal et al., 2020). Consecutively chromosome 3B and 3D consensus maps and solid stem LS mean obtained for each genotype were also used for QTL analysis using QTL Cartographer (version 1.6). Genotyping, linkage mapping and quantitative trait loci analysis Two-locus QTL analysis was carried out to map epistatic QTLs using QTLNetwork (version 2.0) (Yang et al., 2008) at default parameters. Linkage maps illustrating homologous relationships across populations/consensus map, QTL positions and LOD contour across environments for line solid stem score (S) and internode score (I) were drawn using MapChart software (version 2.32) (Voorrips, 2002). ‘Sadash/P2711’ population genotyping data was transformed into the mapping data format “ABH” (A = Sadash, B = P2711, H = heterozygous). Subsequently, the JoinMap® 4.0 program (Van Ooijen, 2006) was used for pre-mapping of the SNPs with respect to the veriﬁcation of segregation patterns, the initial formation of linkage groups (LGs) and the preliminary position of the markers on the chromosomes using the default grouping settings and the mapping algorithm ML (maximum likelihood). The ﬁnal map position of the markers and the genetic distances between the markers were then optimized manually regarding the number of crossovers (as low as possible) and the length of the linkage group (as short as possible) using the ABH mapping data ﬁle in Excel and MapManager QTX (parameters: linkage evaluation - double haploid, search linkage criterion - p=0.05, map function - Kosambi, cross-type - line cross) (Manly et al., 2001). Some markers were eliminated from the ﬁnal mapping if they spread the linkage group due to too many crossovers. Stem solidness assessment Solid stem assessment in all trial experiments, except in the New Zealand ﬁeld trial, was done following (Depauw and Read, Frontiers in Plant Science 04 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 Dhariwal et al. Genotyping, linkage mapping and quantitative trait loci analysis were corrected after re-scoring genotyping data manually in Microsoft Excel followed by recalculating the linkage map using MapDisto software (version 1.7.7.011) (Lorieux, 2012). Two or more linkage groups generated for the same chromosome were tried to merge into a single linkage group using less stringent cut-off values (r max > 0.3) using MapDisto. Linkage mapping of the ‘AAC Cameron/P2711’ population was done as described earlier (Farzand et al., 2022). Four consensus maps were generated for each of the chromosomes 3B and 3D separately utilizing individual LG maps of three populations using the R package LPmerge (version 1.7) (Endelman and Plomion, 2014). Final consensus maps for chromosomes 3B and 3D were selected based on the lowest mean and standard deviation for root-mean-squared error (RMSE) between the consensus map and the linkage maps. Whole genome linkage map plots illustrating SNP density were drawn using the R package LinkageMapView (version 2.1.2) (Ouellette et al., 2018). For isolation of DNA from leaf samples, four seeds were sown from each genotype in 96 cell seed planting trays (parents and checks repeated over all trays) in a soil mixture of Turface (9.07 kg), Peat Moss (0.907 kg) and Vermiculite (0.06 m3). Leaf tissue samples were collected from 10 days old seedlings followed by isolation of DNA using DNeasy 96 Plant Kit (Qiagen Inc., Valencia, CA, USA) following Dhariwal et al. (2018). Quant-iT™PicoGreen® dsDNA Assay Kit (Thermo Fisher Scientiﬁc Inc., Bartlesville, OK, USA) was used to quantify DNA samples followed by dilutions of DNA samples to 50 ng/µl. ‘Sadash/P2711’ population, its parents and checks were genotyped using Wheat 15K Illumina Inﬁnium SNP array from TraitGenetics GmbH, Germany. ‘AAC Innova/P2711’ and ‘AAC Cameron/P2711’ populations, their parents and checks were genotyped using a 90K Illumina Inﬁnium array (Wang et al., 2014). Single nucleotide polymorphism (SNP) genotyping data were analyzed using Genotyping module of GenomeStudio software (Illumina Inc., CA, USA) as described previously (Dhariwal et al., 2020). Brieﬂy, high quality SNPs were selected from the list of all SNPs evaluated for genotyping using following ﬁlters in GenomeStudio (i) Call Freq: >0.50, (ii) Minor Allele Freq: >0.03, (iii) AA Freq:!=1, (iv) AB Freq:!=1, and (v) BB Freq:!=1. Monomorphic and SNPs that deviated signiﬁcantly from the 1:1 ratio were removed by comparing the parental proﬁle and using the c2 test, respectively. Frontiers in Plant Science Assessment of syntenic relationship, assignment of QTL genomic intervals and mapping of candidate gene The LS mean of solid stem rating values for P2711 was 4.92 or ~5.0 across environments, while it was ~1.5 for Sadash and ~1.6 for ‘AAC Innova’. The population mean values of solid stem rating ranged from 2.95 in LTGH19 to 3.17 in LTFD19 for the ‘Sadash/ P2711’ population and 3.03 in LTGH15 to 3.22 in LTFD15 for the ‘AAC Innova/P2711’ population. Histograms showed either slightly skewed distribution towards more excellent stem solidness or binomial distribution for both populations across environments, except Lethbridge 2019 greenhouse experiments (LTGH19) for the ‘Sadash/P2711’ population and New Zealand, 2013 ﬁeld trial (NZFD13) for ‘AAC Innova/P2711’ population (Figures 2, 3). Additionally, a broader range of genotypes was observed across environments in both populations (Figures 2, 3). Strong genotypic effect and high heritability of pith/stem solidness expression across environments along with skewed (towards more excellent solidness)/binomial distribution in both populations indicate the role of P2711 derived major solid stem genes/QTLs and no evidence for transgressive segregation (Lanning et al., 2006). However, a signiﬁcant environmental effect also suggests the involvement of some minor and unstable QTLs. Similar to the BLAST search for syntenic relationships, probe sequences of all the SNPs (wheat 90K iSelect SNP array and markers unique to 15K Wheat Inﬁnium array) that were mapped into QTL intervals in this study were also BLAST searched against the Chinese Spring wheat reference genome sequence (IWGSC RefSeq v2.1) to identify the QTL genomic intervals on wheat chromosomes. cDNA sequence of durum solid stem gene TdDof (TRITD3Bv1G280530.1) was also BLAST searched against the wheat reference genome sequences to identify common wheat homologue of TdDof. Haplotype graphical genotypes and boxplot distributions were estimated involving 3B linkage maps, genotypic data and stem solidness score of recombinant lines, parental and check cultivars belonging to ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations using software package GGT 2.0 (Van Berloo, 2008), and R packages ggpubr (Kassambara, 2016) and rstatix (Kassambara, 2020). Statistically signiﬁcant differences among loci and their combinations were calculated by pairwise Games-Howell test using packages ggstatsplot (Patil, 2021) and tidyverse (Wickham et al., 2019) in R (version 4.0.3) (R Core Team, 2013). Correlation coefﬁcients (r) for the solid stem rating between any pair of environments were quite high and highly signiﬁcant in the ‘Sadash/P2711’ population (Figure 2) and moderate to very high but highly signiﬁcant in the ‘AAC Innova/P2711’ population (Figure 3). Assessment of syntenic relationship, assignment of QTL genomic intervals and mapping of candidate gene Markers belonging to the ‘AAC Innova/P2711’ population were assigned to linkage groups (LGs) using MSTmap (version 2.0) software (Wu et al., 2008) at default grouping settings and mapping function Kosambi (Kosambi, 1943) following Dhariwal and Randhawa (2022) and Farzand et al. (2021). Wheat chromosomes to LGs were assigned using the wheat 90K consensus SNP map (Wen et al., 2017). Double recombinants Syntenic relationships of 3B and 3D linkage maps of ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations with physical maps of chromosomes 3B and 3D (reference genome assembly PGSBv2.1) of Canadian solid-stemmed spring wheat Frontiers in Plant Science 05 frontiersin.org frontiersin.org 10.3389/fpls.2022.1035620 Dhariwal et al. cultivar ‘CDC Landmark’ (pedigree: Unity/Waskada//Alsen/ Superb) was assessed using BLAST search following Dhariwal et al. (2021) and Dhariwal and Randhawa (2022). Brieﬂy, chromosome 3B (GenBank accession: LR877316.1) and 3D (GenBank accession: LR877317.1) reference sequences of cv ‘CDC Landmark’ were downloaded from the National Center for Biotechnology Information (NCBI), which were BLAST searched (with at least 99% identity and 100% query coverage) utilizing probe sequences of SNP markers [wheat 90K iSelect SNP array markers (Wang et al., 2014) and markers unique to 15K Wheat Inﬁnium array (Soleimani et al., 2020)], which were mapped to 3B and 3D linkage groups of ‘Sadash/P2711’ and/or ‘AAC Innova/P2711’ populations, using NCBI standalone BLAST program (Altschul et al., 1990). Genomic coordinates identiﬁed from BLAST results were used to decipher syntenic relationships of 3B and 3D linkage maps of ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations with ‘CDC Landmark’ genome using Windows software program Strudel (version 1.15.08.25) (https://ics.hutton.ac.uk/strudel/). solidness was extremely low in the ‘AAC Cameron/P2711’ RIL population across environments, perhaps due to one or more reasons such as environmental effects (Weiss and Morrill, 1992; Cook et al., 2004), the residual heterozygosity present in RILs (Platt, 1941; Lanning et al., 2006), and unintended selection occurred during early generations. Thus, the phenotypic data of this population was not utilized for further analysis, however, genotypic data and the genetic map (Farzand et al., 2022) generated from ‘AAC Cameron/P2711’ at an advanced generation were utilized for the generation of chromosomes 3B and 3D speciﬁc consensus maps. The ANOVA of solid stem score for both ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations revealed highly signiﬁcant effects of genotypes as well as environments but not of genotype x environment interaction (Supplementary Table S2). The estimated broad-sense heritability (H2) values were also strong (0.78 for ‘Sadash/P2711’ and 0.69 for ‘AAC Innova/P2711’). Assessment of syntenic relationship, assignment of QTL genomic intervals and mapping of candidate gene For the ‘Sadash/P2711’ population, Lethbridge 2019 greenhouse experiment showed relatively lower correlations with other environments (Figure 2). Similarly, the 2013 Lincoln, New Zealand experiment showed relatively lower correlations with other environments for the ‘AAC Innova/P2711’ population (Figure 3). Lower correlations between any environment pair than the broad-sense heritability values (0.78 for ‘Sadash/P2711’ and 0.69 for ‘AAC Innova/ P2711’ populations) further suggest the existence of strong environmental inﬂuence (Gondo et al., 2007) on stem solidness which has also been reported previously for solid- Frontiers in Plant Science Phenotypic observations: Solid stem expression was high in doubled haploid lines Phenotypic data analysis revealed substantial phenotypic variability for expression of stem solidness among the parents, check cultivars and DH lines across all environments for ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations (Figures 1–3; Supplementary Table S1). Expression of stem Frontiers in Plant Science 06 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 1 A representative picture of longitudinal dissection of stems (main tillers) of the three parental cultivars. Hollow stem cultivars Sadash and ‘AAC Innova’ are respectively shown on the left-hand side and in the middle while solid stem cultivar P2711 is on the right-hand side. Wheat stem sawﬂy (WSS) larvae detected in the stem of Sadash are shown by blue arrows while the girdled section ﬁlled with larva frass in both hollow stem cultivars Sadash and ‘AAC Innova’ are shown by orange arrows. All internodes of the stem of P2711 are completely ﬁlled with pith and have no sign of WSS damage. The picture was taken from mature plant stems collected from an irrigated ﬁeld in Lethbridge, AB on September 1, 2022. 4,959 SNP markers were found polymorphic between two parents of the ‘Sadash/P2711’ population, of which, 4,837 were mapped to 24 linkage groups in that population (Figure 4; Supplementary Table S3). A total of 13,425 SNPs were found polymorphic between two parents of the ‘AAC Innova/P2711’ population, of which, 8,930 high-quality SNPs were successfully mapped to 23 linkage groups (Figure 5; Supplementary Table S4). Map of ‘AAC Cameron/P2711’ has been described and published previously (Farzand et al., 2022). Brieﬂy, a total of 8,915 markers were incorporated into 29 linkage groups constructed using this population. stem phenotype in common wheat DH population ‘Rampart/ Jerry’ (Cook et al., 2004). Irrespective of environmental effects, DH solid stem data was of high quality and found suitable for identifying QTLs in these populations. Frontiers in Plant Science High-density linkage maps Frequency distribution histograms with distribution curve (blue line) for solid stem score of DHs grown at Lethbridge, Alberta in the ﬁeld in 2014 (LTFD14S) and 2019 (LTFD19S) and greenhouse in 2019 (LTGH19S), as well as pooled data (average of all three environments; S) are shown on main diagonal. The means of the parental genotypes, Sadash (hollow-stemmed) and P2711 (solid-stemmed), are indicated by red and blue dots, respectively, beneath the frequency distribution plots. Scatterplots with regression lines, linear (blue) and exponential (red), for each environment are shown on the left side of the main diagonal. Orange dots on scatterplots represent the solid stem score of DHs. Correlation coefﬁcients (r) between each pair of environments, and each environment and the pooled data, are displayed on the right side of the main diagonal. Color intensity (light red to dark red) on r boxes indicates the depth of association between environments under evaluation. p ≤0.001. ‘Sadash/P2711’ map spanned a map distance of 2953.9 cM (on average 123.08 cM per LG or 140.66 cM per chromosome) with an average marker interval of 2.05 cM and an average SNP distance of 0.61 cM (Figure 4; Supplementary Table S5). Similarly, the ‘AAC Innova/P2711’ map spanned a map distance of 3010.65 cM (on average 130.9 cM per LG or 143.36 cM per chromosome) with an average marker interval of 1.12 cM and an average SNP distance of 0.34 cM (Figure 5; Supplementary Table S5). The total map lengths of linkage groups for sub-genomes A, B, and D were 1208.5, 1124.2, and 621.2, respectively, for the ’Sadash/P2711’ population; similarly, sub-genomes A, B, and D map lengths were 1315.65, 1140.68, and 554.32, respectively, for the ‘AAC Innova/P2711’ population. These results showed that where genome A had the highest SNP coverage in both populations, which is similar to genome B, genome D had the lowest coverage with almost half of either A or B genome. In both linkage maps, the D-genome chromosomes are signiﬁcantly underrepresented in comparison with the A- and B-genome chromosomes. We also observed uneven distribution of SNPs along the linkage groups and the formation of SNP clusters in certain regions which led to the formation of large gaps in many linkage groups, particularly in some sub-genome D-speciﬁc chromosomes in both populations (Figures 4, 5). High-density linkage maps It may be recalled that the ‘Sadash/P2711’ population was genotyped using Illumina 15K Wheat SNP array while ‘AAC Innova/P2711’ using Illumina 90K Wheat SNP array. A total of Frontiers in Plant Science 07 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 2 Frequency distribution and correlation scatterplots for solid stem rating of doubled haploid population ‘Sadash/P2711’. Frequency distribution histograms with distribution curve (blue line) for solid stem score of DHs grown at Lethbridge, Alberta in the ﬁeld in 2014 (LTFD14S) and 2019 (LTFD19S) and greenhouse in 2019 (LTGH19S), as well as pooled data (average of all three environments; S) are shown on main diagonal. The means of the parental genotypes, Sadash (hollow-stemmed) and P2711 (solid-stemmed), are indicated by red and blue dots, respectively, beneath the frequency distribution plots. Scatterplots with regression lines, linear (blue) and exponential (red), for each environment are shown on the left side of the main diagonal. Orange dots on scatterplots represent the solid stem score of DHs. Correlation coefﬁcients (r) between each pair of environments, and each environment and the pooled data, are displayed on the right side of the main diagonal. Color intensity (light red to dark red) on r boxes indicates the depth of association between environments under evaluation. p ≤0.001. FIGURE 2 Frequency distribution and correlation scatterplots for solid stem rating of doubled haploid population ‘Sadash/P2711’. Frequency distribution histograms with distribution curve (blue line) for solid stem score of DHs grown at Lethbridge, Alberta in the ﬁeld in 2014 (LTFD14S) and 2019 (LTFD19S) and greenhouse in 2019 (LTGH19S), as well as pooled data (average of all three environments; S) are shown on main diagonal. The means of the parental genotypes, Sadash (hollow-stemmed) and P2711 (solid-stemmed), are indicated by red and blue dots, respectively, beneath the frequency distribution plots. Scatterplots with regression lines, linear (blue) and exponential (red), for each environment are shown on the left side of the main diagonal. Orange dots on scatterplots represent the solid stem score of DHs. Correlation coefﬁcients (r) between each pair of environments, and each environment and the pooled data, are displayed on the right side of the main diagonal. Color intensity (light red to dark red) on r boxes indicates the depth of association between environments under evaluation. p ≤0.001. FIGURE 2 Frequency distribution and correlation scatterplots for solid stem rating of doubled haploid population ‘Sadash/P2711’. Frontiers in Plant Science frontiersin.org High-density linkage maps Orange dots on scatterplots represent the solid stem score of DHs. Correlation coefﬁcients (r) between each pair of environments, and each environment and the pooled data, are displayed on the right side of the main diagonal. Color intensity (light red to dark red) on r boxes indicates the depth of association between environments under evaluation. p ≤0.001. are in agreement with the previous genetic maps (Wang et al., 2014; Wen et al., 2017; Dhariwal et al., 2018; Dhariwal et al., 2020; Farzand et al., 2021; Farzand et al., 2022). Moreover, maps constructed in this study cover all 21 wheat chromosomes and provides enough coverage to dissect the genetic variation of stem solidness present in these populations. ‘AAC Innova/P2711’, respectively) but the lowest number to group 4 chromosomes (335 SNPs) in ’Sadash/P2711’ population and group 5 chromosomes (752) in ‘AAC Innova/P2711’ population. Likewise, the highest marker densities were reported on group 1 chromosomes in both populations (0.35 and 0.21 cM/SNP, respectively in the ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations). Interestingly, a total of 74 SNPs belonging to the ‘Sadash/P2711’ population and 317 to the ‘AAC Innova/P2711’ population mapped in this study represent new mapped markers (Supplementary Tables S3, S4) which were not present on either wheat consensus map (Wen et al., 2017) or previous wheat maps used for solid stem studies (Nilsen et al., 2017). High-density linkage maps These variations are perhaps due to (i) the lower number of analyzed markers on genome D or other genomes, (ii) similarity of genomic regions where the parents are identical by descent, and (iii) exome speciﬁcity of the mapped SNPs (Dhariwal et al., 2018). Similar results have also been reported previously (Dhariwal et al., 2018; Dhariwal et al., 2020). On the other hand, the highest number of SNPs were mapped to wheat homoeologous group 1 chromosomes in both populations (863 and 2102 SNPs in ‘Sadash/P2711’ and Frontiers in Plant Science frontiersin.org 08 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 3 Frequency distribution and correlation scatterplots for solid stem rating of doubled haploid population ‘AAC Innova/P2711’. Frequency distribution histograms with distribution curve (blue line) for solid stem score of DHs grown in New Zealand ﬁeld trial in 2013 (NZFD13S), at Lethbridge, Alberta in the greenhouse in 2015 (LTGH15S), the ﬁeld in 2015 (LTFD15S) and 2019 (LTFD19S), as well as pooled data (average of all three environments; S) are shown on main diagonal. The means of the parental genotypes ‘AAC Innova’ (hollow-stemmed) and P2711 (solid- stemmed) are indicated by red and blue dots, respectively, beneath the frequency distribution plots. Scatterplots with regression lines, linear (blue) and exponential (red), for each environment are shown on the left side of the main diagonal. Orange dots on scatterplots represent the solid stem score of DHs. Correlation coefﬁcients (r) between each pair of environments, and each environment and the pooled data, are displayed on the right side of the main diagonal. Color intensity (light red to dark red) on r boxes indicates the depth of association between environments under evaluation. *p ≤0.001. FIGURE 3 Frequency distribution and correlation scatterplots for solid stem rating of doubled haploid population ‘AAC Innova/P2711’. Frequency distribution histograms with distribution curve (blue line) for solid stem score of DHs grown in New Zealand ﬁeld trial in 2013 (NZFD13S), at Lethbridge, Alberta in the greenhouse in 2015 (LTGH15S), the ﬁeld in 2015 (LTFD15S) and 2019 (LTFD19S), as well as pooled data (average of all three environments; S) are shown on main diagonal. The means of the parental genotypes ‘AAC Innova’ (hollow-stemmed) and P2711 (solid- stemmed) are indicated by red and blue dots, respectively, beneath the frequency distribution plots. Scatterplots with regression lines, linear (blue) and exponential (red), for each environment are shown on the left side of the main diagonal. Frontiers in Plant Science Two major solid stem loci derived from P2711 Whole genome QTL analyses identiﬁed that solid stem expression in both populations was mostly modulated by two major and stable loci mapped to chromosomes 3B (QSst.lrdc-3B) and 3D (QSst.lrdc-3D) in addition to some minor loci (Table 1; Figures 6, 7; Supplementary Table S6) as well as epistasis effect Where the distributions of mapped markers over the linkage groups of all populations are satisfactory, the map length and the number of markers of linkage groups constructed in this study Frontiers in Plant Science 09 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 4 ‘Sadash/P2711’ linkage map constructed using single nucleotide polymorphism (SNP) markers from 15K wheat SNP array. FIGURE 4 ‘Sadash/P2711’ linkage map constructed using single nucleotide polymorphism (SNP) markers from 15K wheat SNP array. (Supplementary Table S7). The two major QTLs are discussed ﬁrst in greater detail followed by others below. explain as much as 76% (Cook et al., 2004) to 78% (Nilsen et al., 2017) of phenotypic variation for whole stem rating in common wheat. Cook et al. (2017) demonstrated that all WSS-resistant common wheat North American cultivars, except ‘Conan’, evaluated in their study had the same haplotype at the major 3BL locus derived from ‘S-615’ but this haplotype was not found in the solid-stem tetraploid landraces. Recently, using more detailed analysis, Nilsen et al. (2020) reported that a single gene is responsible for stem solidness at the 3B locus across durum and common wheat cultivars, and perhaps it originated from a common genetic progenitor, either pre- or post- domestication. Although the exact source of stem solidness at the 3B locus in P2711 is unknown, P2711 also most likely derived its stem solidness from the same progenitor as other durum and common wheat cvs. since both P2711 and the durum landrace ‘Golden Ball’ originated from South Africa. QSst.lrdc-3B mapped on the distal region of the long arm of chromosome 3B was detected in both populations across all environments as well as in pooled data. Moreover, while it was detected using whole stem solidness data, it was also detected for all 4 upper internodes rated in this study. Using population- speciﬁc maps, we identiﬁed that this QTL was derived from the solid-stem parent P2711 and explained up to 64.0% and 52% of the phenotypic variation (PV) for whole stem solidness and 77.0% and 58.0% of PV for internodes in ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations, respectively (Table 1). frontiersin.org Two major solid stem loci derived from P2711 The P2711 allele at this locus had an internode additive effect up to 1.33 in ‘Sadash/P2711’ population and up to 1.18 in the ‘AAC Innova/P2711’ population (Table 1) which increased stem solidness by around 24.0 - 27.0%. Major QTLs have been previously repeatedly mapped to the same overlapping region as QSst.lrdc-3B in common wheat cultivars (e.g., Canadian and US cvs. Choteau, Lillian Rescue, Rampart and Scholar all derived from Portuguese landrace S-615 and cv. with unknown solid stem resource like US cv. Conan) and to the distal region of chromosome 3B of durum wheat (e.g., Canadian cvs. derived from German cv. Biodur and South African landrace Golden Ball) (Cook et al., 2004; Lanning et al., 2006; Houshmand et al., 2007; Sherman et al., 2010; Talbert et al., 2014; Varella et al., 2015; Nilsen et al., 2017). Similar to P2711-derived 3B QTL, previously identiﬁed QTLs in this region were also reported to QSst.lrdc-3D is another very strong major and stable QTL derived from the male parent P2711 and explained up to 40.0% and 46.0% of the phenotypic variation for whole stem solidness and up to 52.0% and 56.0% of PV for internodes in ‘Sadash/ P2711’ and ‘AAC Innova/P2711’ populations, respectively (Table 1). Like QSst.lrdc-3B, QSst.lrdc-3D was also mapped on the distal region of the long arm of its chromosome 3D and detected in both populations across all environments as well as in pooled data for whole stem solidness and 4 internodes rating (Table 1; Figure 7; Supplementary Table S6). The P2711 allele at this locus had an internode additive effect up to 1.56 in ‘Sadash/ Frontiers in Plant Science 10 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 5 ‘AAC Innova/P2711’ linkage map constructed using single nucleotide polymorphism (SNP) markers from 90K wheat SNP array. FIGURE 5 ‘AAC Innova/P2711’ linkage map constructed using single nucleotide polymorphism (SNP) markers from 90K wheat SNP array. P2711’ and up to 1.00 in the ‘AAC Innova/P2711’ population (Table 1) which increased stem solidness by around 20.0 - 31.0%. A secondary QTL (Qss.msub-3DL) has also been previously mapped to chromosome arm 3DL in common wheat cultivar Choteau (Lanning et al., 2006). Interestingly, both Choteau and P2711 are hard red spring wheat lines and possess the solid stem locus on the distal region of chromosome arm 3DL. However, while Choteau QTL explained only 31.0% of phenotypic variation, Choteau also does not share its immediate parentage with P2711. frontiersin.org Two major solid stem loci derived from P2711 Moreover, we found that Qss.msub-3DL’s closely associated microsatellite marker Xgwm645 (Lanning et al., 2006) is physically located proximal (chromosome position 454946571) to QSst.lrdc-3D’s closely linked SNP marker IWB52401 (chromosome position 609087492) on chromosome 3D of cv ‘CDC Landmark’. Since the original source of stem solidness in P2711 is not known and differences in positions of QSst.lrdc-3D and Qss.msub-3DL may be due to shifts in QTL positions as a result of differences in phenotyping or genotyping, it remains to be determined whether QSst.lrdc-3D and Qss.msub-3DL are the same or represent different loci. However, the differences in explained PV of QSst.lrdc-3D and Qss.msub-3DL indicate that the P2711 most likely carries at least a different allele at this locus. Future haplotype study of two genotypes, Choteau and P2711, using all closely linked and gene-based markers of QSst.lrdc-3D and Qss.msub-3DL may help in establishing a deﬁnitive relationship between the two identiﬁed QTLs. To identify additional markers at these QTLs as well as markers common between two populations, we constructed consensus maps for chromosomes 3B and 3D using the mapping data of all three individual populations used in this study. Chromosome 3B consensus map spans a total map length of 179.836 cM, while the 3D map spans 23.147 cM (Supplementary Tables S8, S9). These Frontiers in Plant Science 11 frontiersin.org frontiersin.org S Int. S Int 64.0 77.0 40.0 52.0 5.0 5.0 6.0 11.0 4.0 6.0 3.0 2.0 2.0 6.0 9.0 9.0 3.0 8.0 2.0 na 52.0 58.0 46.0 56.0 1.0 na 2.0 na 3.0 3.0 na 4.0 na 5.0 na 3.0 4.0 6.0 1.0 2.0 2.0 2.0 2.0 4.0 na 2.0 TL in any single environ 9; and LTFD19, Lethbri d/or internode solidness detected QTL; S, stem so ucleotide polymorphism m al map/genome, respect Minor solid-stem loci In addition to above two major stable QTLs, we detected a number of minor QTLs on chromosomes 1B (‘AAC Innova/ P2711’ population), 1D (‘Sadash/P2711 population’), 2A (both populations), 2B and 2D (‘AAC Innova/P2711’ population), 4A (both population), 4D (‘AAC Innova/P2711’ population), 5A (both populations), 5B and 5D (‘AAC Innova/P2711’ population), 6B (‘Sadash/P2711’ population) and 7B (both population) (Table 1). Most of these minor loci explained <5% of phenotypic variation for whole stem solidness except QSst.lrdc-1D, QSst.lrdc-2A and QSst.lrdc-5A.2 from the ‘Sadash/P2711’ population, which respectively explained 5%, 6% and 9% phenotypic variation. Moreover, most of these loci were detected in one environment/ one environment with pooled data or just pooled data, except QSst.lrdc-2A and QSst.lrdc-6B from the ‘Sadash/P2711’ population and QSst.lrdc-2B from ‘AAC Innova/P2711’ population. Except for QTLs QSst.lrdc-5A.2, QSst.lrdc-6B, and QSst.lrdc-7B of ‘Sadash/ P2711’ population and QSst.lrdc-2B and QSst.lrdc-5D of ‘AAC Innova/P2711’ population, solid stem allele at all loci were derived from P2711 in both populations. Solid stem alleles at remaining ﬁve loci were derived from the hollow-stemmed parent of respective population. On the other hand, QSst.lrdc-2A of ‘Sadash/P2711’ and QSst.lrdc-2A.2 of ‘AAC Innova/P2711’ are supposed to be the same locus as they shared a common genomic location on the wheat reference genome (Supplementary Tables S11, S12). Similarly, QSst.lrdc-4A.2 of ‘Sadash/P2711’ and QSst.lrdc-4A of ‘AAC Innova/P2711’ belongs to the same locus on chromosome 4A, in addition to QSst.lrdc-5A.2 of ‘Sadash/P2711’ QSst.lrdc-3D is homeologous to QSst.lrdc-3B QSst.lrdc-3D is homeologous to QSst.lrdc-3B Synteny analysis of chromosomes 3B and 3D linkage maps of both populations with the 3B physical map of ‘CDC Landmark’, a Canadian solid stem wheat cultivar, showed the consistency of marker order between the genetic and physical maps except for a few SNPs (Supplementary Figure S1). A similar syntenic relationship was observed when we compared the 3B and 3D linkage maps of both populations with the 3D physical map of ‘CDC Landmark’ (Supplementary Figure S2). Particularly, the order of SNP markers on 3B linkage groups of both populations and 3B and 3D physical maps of ‘CDC Landmark’ was highly consistent (Supplementary Figure S2). However, we observed that a lower recombination rate or similarity of 3D chromosomes of population parents in both crosses (‘Sadash/P2711’ and ‘AAC Innova/P2711’), which largely seems to be identical by descent, leads to the low-resolution mapping of markers on this chromosome. This also resulted in some discrepancies between the marker order of 3D linkage maps of two crosses and the physical map (Supplementary Figure S2). Interestingly, QSst.lrdc- 3B and QSst.lrdc-3D from both populations overlap the same genomic region, which also possesses the wheat homologue of durum solid stem gene TdDof, on the 3B physical map of ‘CDC Landmark’ (Supplementary Figure S1). Similarly, both major solid stem loci identiﬁed in this study overlap the same genomic region on the 3D physical map of ‘CDC Landmark’ (Supplementary Figure S2) irrespective of minor marker order discrepancies. Our results indicate that both solid stem loci identiﬁed in this study are homeologous to each other. Two major solid stem loci derived from P2711 SNP ID cM bp 3 64.0 77.0 LTFD14, LTFD19, LTGH19, C S, I1 – I4 IWB11701 184.1 843247610 P2711 6 40.0 52.0 LTFD14, LTFD19, LTGH19, C S, I1 – I4 TG3069 20.4 603821560 P2711 4 5.0 5.0 LTGH19 S, I3 IWA3753 7.8 3051756 P2711 3 6.0 11.0 LTFD14, LTFD19, C S, I1 – I3 IWB11289 53.9 64102080 P2711 2 4.0 6.0 LTFD14 S, I2 IWB8008 0.0 3365864 P2711 5 3.0 2.0 C S, I2 IWB46934 50.0 487314533 P2711 5 2.0 6.0 LTGH19, C S, I2 IWB36264 76.4 485652482 P2711 9.0 9.0 LTGH19, C S, I2 – I4 IWB47006 166.8 609465091 HP 3.0 8.0 LTFD14, LTGH19 S, I1, I3 IWB60190 92.7 657946476 HP 2.0 na C S IWB6919 11.9 3541540 HP 8 52.0 58.0 NZFD13, LTGH15, LTFD15, LTFD19, C S, I1 – I4 IWA1756 168.7 843250756 P2711 0 46.0 56.0 NZFD13, LTGH15, LTFD15, LTFD19, C S, I1 – I4 IWB52401 16.5 611013692 P2711 2 1.0 na LTGH15 S IWB26730 33.2 10355943 P2711 6 2.0 na LTFD19 S IWB10662 16.0 36651065 P2711 1 3.0 3.0 LTGH15, C S, I1 – I4 IWA3194 58.6 76368584 P2711 na 4.0 LTGH15, LTFD15, LTFD19 I2 – I3 IWA5410 56.1 52005347 HP 2 na 5.0 LTFD15 I1 IWB64223 141.0 640832761 P2711 7 na 3.0 LTGH15 I1 IWB1356 24.6 345813273 P2711 4 4.0 6.0 LTFD15 S, I1 IWB12286 16.8 455964578 P2711 6 1.0 2.0 LTGH15 S, I2 – I3 IWB71951 67.0 482845859 P2711 4 2.0 2.0 LTGH15 S, I2 – I4 IWB72268 55.4 459560328 P2711 2.0 4.0 LTGH15 S, I1, I3 IWB63558 60.8 465885796 HP 7 na 2.0 LTGH15 I3 IWB51995 9.0 13398372 P2711 QTL in any single environment (NZFD13, New Zealand Field Trial 2013; LTFD14, Lethbridge Field Trial 2014; LTGH15, Lethbridge Greenhouse Experiment 2015; 019; and LTFD19, Lethbridge Field Trial 2019) or pooled/combined (C) data of stem/internode solidness; Intervalmax, QTL interval (cM) calculated using markers nd/or internode solidness dataset(s); LOD, logarithm of the odds score for a detected QTL; LODmax, Additive effectmax and %R2 max, highest score reported in any single a detected QTL; S, stem solidness; Int., internode solidness; S and/or Int., QTL detected for stem solidness (S) and/or internode (I) score; I1, internode 1/peduncle; I2, nucleotide polymorphism marker identity based on 90K Wheat SNP array or 15K Wheat SNP array (*); cM, centimorgan; bp, base pairs; cM and bp positions are based ical map/genome, respectively; QTLs common in both populations; Donor allele: P2711 – solid stem parent, HP – hollow stem parent Sadash in ‘Sadash/P2711’ Dhariwal et al. Two major solid stem loci derived from P2711 Interestingly, TaDof, a homolog of the durum TdDof gene, was located outside (at 843.64 Mb) this region on chromosome 3B. SNP marker RAC875_c48860_106 (IWB58481), which was genetically mapped distal to this 1.1 Mb ‘3B solid stem segment’ on ‘AAC Innova/P2711’ 3B map, had an ‘AAC Innova’ type allele in cv. ‘Golden Ball’ and was located physically at 843.94 Mb on Chinese Spring chromosome 3B. Since, the position of recombination breakpoint between markers co-segregating in the ‘3B solid stem segment’ and the SNP marker RAC875_c48860_106 is not known due to genetic type of data, which may be anywhere between 843.25 and 843.94 Mb, it remains to be determined whether TaDof co-segregate with ‘3B solid stem segment’ or not. Two major solid stem loci derived from P2711 10.3389/fpls.2022.1035620 10.3389/fpls.2022.1035620 maps have higher resolution (map density: 0.16 and 0.04 cM/SNP, respectively, for chromosomes 3B and 3D) and higher marker coverage than individual population-speciﬁc maps. QTL analysis using these maps identiﬁed the same solid stem loci as detected using population-speciﬁc maps but with higher conﬁdence and additional markers (Supplementary Table S10). We further observed that QSst.lrdc-3B explained up to 93.0% of phenotypic variation for whole stem solidness and 78.0% for internodes, while QSst.lrdc-3D explained up to 46.0% of phenotypic variation for whole stem solidness and 67.0% for internodes (Supplementary Table S10). We also found several common markers between ‘Sadash/P2711’ and ‘AAC Innova/P2711’ populations for each of the two chromosomes, 3B and 3D (Figures 6, 7). Selection for the solid stem alleles on chromosome arms 3BL and 3DL using these common markers will be more useful than population speciﬁc markers for the deployment of stem solidness in different genetic backgrounds to develop new cultivars. (Figure 8). We further narrowed this region using haplotype data of the ‘AAC Innova/P2711’ population and cvs Lillian and ‘Golden Ball’, which located 3B solid stem underlying gene between 842.14 and 843.25 Mb on chromosome 3B (hereinafter referred to as ‘3B solid stem segment’) of Chinese Spring (Figure 8). We found that both Lillian and ‘Golden Ball’ share common alleles in this genomic region. Interestingly, TaDof, a homolog of the durum TdDof gene, was located outside (at 843.64 Mb) this region on chromosome 3B. SNP marker RAC875_c48860_106 (IWB58481), which was genetically mapped distal to this 1.1 Mb ‘3B solid stem segment’ on ‘AAC Innova/P2711’ 3B map, had an ‘AAC Innova’ type allele in cv. ‘Golden Ball’ and was located physically at 843.94 Mb on Chinese Spring chromosome 3B. Since, the position of recombination breakpoint between markers co-segregating in the ‘3B solid stem segment’ and the SNP marker RAC875_c48860_106 is not known due to genetic type of data, which may be anywhere between 843.25 and 843.94 Mb, it remains to be determined whether TaDof co-segregate with ‘3B solid stem segment’ or not. (Figure 8). We further narrowed this region using haplotype data of the ‘AAC Innova/P2711’ population and cvs Lillian and ‘Golden Ball’, which located 3B solid stem underlying gene between 842.14 and 843.25 Mb on chromosome 3B (hereinafter referred to as ‘3B solid stem segment’) of Chinese Spring (Figure 8). We found that both Lillian and ‘Golden Ball’ share common alleles in this genomic region. Frontiers in Plant Science Haplotype analysis located QSst.lrdc-3B to a 1.1 Mb region on chromosome 3B FIGURE 6 FIGURE 6 The linkage maps of chromosome 3B deciphering the logarithm of odds (LOD) curve plots for major solidness (stem, S and internode, I) quantitative trait locus (QTL) QSst.lrdc-3B detected using composite interval mapping from ‘Sadash/P2711’ (3B-SP) and ‘AAC Innova/P2711’ (3B-IP) doubled haploid populations in each of the trial environments and pooled data along with a chromosome 3B consensus map (3B-C) in middle. Positions of markers are shown as lines on linkage group bars and a scale in cM on the left of the graph. SNP markers common between the two linkage groups and the consensus map are shown by SNP IDs in green font, while markers common between a population map and the consensus map are shown in blue (for ‘Sadash/P2711’ markers) or dark pink (for ‘AAC Innova/P2711’ markers) color. Relationships among linkage groups and consensus map are shown using gray color connections for each common marker. QTL intervals are shown using a different colored segment of linkage group bars, while QTL conﬁdence intervals for each environment/trait combination are shown using different colored bars against LOD curve plots. LTFD14(I1-I4, and S), LTFD19(I1-I4, and S), LTGH19(I1-I4, and S), and single letter (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in 2014, 2019, greenhouse experiment in 2019 and pooled data obtained from Lethbridge, Alberta location for whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘Sadash/P2711’ population. Similarly, NZFD13S, LTFD15(I1-I4, and S), LTGH15(I1-I4, and S), LTFD19(I1-I4, and S), and single letters (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in New Zealand in 2013, Lethbridge ﬁeld trials in 2015, Lethbridge greenhouse experiments in 2015, Lethbridge ﬁeld trial in 2019 and pooled data obtained from Lethbridge, Alberta location for the whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘AAC Innova/P2711’ population. Straight lines across the LOD plot represent the LOD signiﬁcance thresholds. Nilsen et al., 2017; Varella et al., 2019). Among these, 1B, 5A, 5B and 5D minor loci were detected in common or durum wheat (Larson and Macdonald, 1959a; Sherman et al., 2010; Varella et al., 2015), while 2A, 3AL, 4A and 6B loci were exclusively detected in durum wheat (Nilsen et al., 2017; Varella et al., 2019). Haplotype analysis located QSst.lrdc-3B to a 1.1 Mb region on chromosome 3B Using haplotype analysis of the ‘Sadash/P2711’ population, we located QSst.lrdc-3B in a chromosomal region between 843.24 and 851.57 Mb on chromosome 3B of Chinese Spring Frontiers in Plant Science 13 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 6 The linkage maps of chromosome 3B deciphering the logarithm of odds (LOD) curve plots for major solidness (stem, S and internode, I) quantitative trait locus (QTL) QSst.lrdc-3B detected using composite interval mapping from ‘Sadash/P2711’ (3B-SP) and ‘AAC Innova/P2711’ (3B-IP) doubled haploid populations in each of the trial environments and pooled data along with a chromosome 3B consensus map (3B-C) in middle. Positions of markers are shown as lines on linkage group bars and a scale in cM on the left of the graph. SNP markers common between the two linkage groups and the consensus map are shown by SNP IDs in green font, while markers common between a population map and the consensus map are shown in blue (for ‘Sadash/P2711’ markers) or dark pink (for ‘AAC Innova/P2711’ markers) color. Relationships among linkage groups and consensus map are shown using gray color connections for each common marker. QTL intervals are shown using a different colored segment of linkage group bars, while QTL conﬁdence intervals for each environment/trait combination are shown using different colored bars against LOD curve plots. LTFD14(I1-I4, and S), LTFD19(I1-I4, and S), LTGH19(I1-I4, and S), and single letter (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in 2014, 2019, greenhouse experiment in 2019 and pooled data obtained from Lethbridge, Alberta location for whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘Sadash/P2711’ population. Similarly, NZFD13S, LTFD15(I1-I4, and S), LTGH15(I1-I4, and S), LTFD19(I1-I4, and S), and single letters (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in New Zealand in 2013, Lethbridge ﬁeld trials in 2015, Lethbridge greenhouse experiments in 2015, Lethbridge ﬁeld trial in 2019 and pooled data obtained from Lethbridge, Alberta location for the whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘AAC Innova/P2711’ population. Straight lines across the LOD plot represent the LOD signiﬁcance thresholds. Frontiers in Plant Science FIGURE 7 FIGURE 7 The linkage maps of chromosome 3D deciphering the logarithm of odds (LOD) curve plots for major solidness (stem, S and internode, I) quantitative trait locus (QTL) QSst.lrdc-3D detected using composite interval mapping from ‘Sadash/P2711’ (3D-SP) and ‘AAC Innova/P2711’ (3D- IP) doubled haploid populations in each of the trial environments and pooled data along with a chromosome 3D consensus map (3D-C) in middle. Positions of markers are shown as lines on linkage group bars and a scale in cM on the left of the graph. SNP markers common between the two linkage groups and the consensus map are shown by SNP IDs in green font, while markers common between a population map and the consensus map are shown in blue (for ‘Sadash/P2711’ markers) or dark pink (for ‘AAC Innova/P2711’ markers) color. Relationships among linkage groups and consensus map are shown using gray color connections for each common marker. QTL intervals are shown using a different colored segment of linkage group bars, while QTL conﬁdence intervals for each environment/trait combination are shown using different colored bars against LOD curve plots. LTFD14(I1-I4, and S), LTFD19(I1-I4, and S), LTGH19(I1-I4, and S), and single letter (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in 2014, 2019, greenhouse experiment in 2019 and pooled data obtained from Lethbridge, Alberta location for whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘Sadash/ P2711’ population. Similarly, NZFD13S, LTFD15(I1-I4, and S), LTGH15(I1-I4, and S), LTFD19(I1-I4, and S), and single letters (and numerals) I1 to I4 and S represent QTL detected in ﬁeld experiments in New Zealand in 2013, Lethbridge ﬁeld trials in 2015, Lethbridge greenhouse experiments in 2015, Lethbridge ﬁeld trial in 2019 and pooled data obtained from Lethbridge, Alberta location for the whole stem and culm internodes I1 (uppermost) to I4 (lowermost) ratings of doubled haploid lines of ‘AAC Innova/P2711’ population. Straight lines across the LOD plot represent the LOD signiﬁcance thresholds. QSst.lrdc-5D which was never mapped earlier. Rest of the minor loci (QSst.lrdc-1D, QSst.lrdc-2A, QSst.lrdc-2A.2, QSst.lrdc-4A.1, QSst.lrdc-4A.2, QSst.lrdc-4D, QSst.lrdc-5A.2, QSst.lrdc-5B, QSst.lrdc-7B) seems to be novel and identiﬁed for the ﬁrst time in this study. minor loci detected in this study, QSst.lrdc-1B may be the same as the 1B locus identiﬁed previously by Sherman et al. (2010) and Varella et al. (2015). FIGURE 7 Similarly, QSst.lrdc-2A.1 and QSst.lrdc-2A.2 may be the same as 2A loci identiﬁed by Nilsen et al. (2017) in durum wheat, QSst.lrdc-5A/QSst.lrdc-5A.1 may be same as 5A locus identiﬁed by Nilsen et al. (2017), and QSst.lrdc-5D may be same as 5D locus identiﬁed by Varella et al. (2015). All chromosome group 5 loci may have also been identiﬁed earlier by Larson and Macdonald (1959a), but their chromosomal locations are precisely detected in this study, particularly Haplotype analysis located QSst.lrdc-3B to a 1.1 Mb region on chromosome 3B While all these loci affect solid stem expression, two (1B and 3AL) of these loci were reported to also inﬂuence or co-segregate with stem cutting along with another trait i.e. heading date (1B) or plant height (3AL) (Sherman et al., 2010; Varella et al., 2015; Varella et al., 2019). Based on genomic locations of markers of and QSst.lrdc-5A of ‘AAC Innova/P2711’ on chromosome 5A (Supplementary Tables S11, S12). These results indicate that the identiﬁed common QTLs can be utilized in different genetic backgrounds (Dhariwal et al., 2021) for improving solid stem as an adaptive trait. Minor QTLs, other than these common QTLs, were population speciﬁc. These unique loci can also be of regional utility (Dhariwal et al., 2021) for solid stem development in wheat. Minor solid stem loci have been previously detected on chromosomes 1B, 2A, 3AL, 4A, 5A, 5B, 5D and 6B (Larson and Macdonald, 1959a; Sherman et al., 2010; Varella et al., 2015; Frontiers in Plant Science 14 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 Frontiers in Plant Science Minor loci were largely detected in upper or middle internodes We observed that while individually, QSst.lrdc-3B contributed maximum stem solidness in both populations, QSst.lrdc-3D either conferred slightly less solidness (in the ‘Sadash/P2711’ population) or almost the same level (statistically insigniﬁcant) of solidness (in ‘AAC Innova/P2711’ population) (Figures 9, 10). Thus, both major loci, QSst.lrdc.3B and QSst.lrdc-3D, were almost equally effective for pith development but together these loci produced a highly solid stem phenotype in both individual wheat populations (Figures 9, 10). Minor loci were largely detected in upper or middle internodes Internode-speciﬁc QTL analysis redetected almost all whole stem solidness QTLs of the ‘Sadash/P2711’ population (except Frontiers in Plant Science 15 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 8 Graphical genotypes of wheat haplotypes (H) of doubled haploid lines (DHLs), hollow (Sadash and ‘AAC Innova’)- and solid (P2711)-stem parent and solid-stem check cultivars (Lillian and ‘Golden Ball’) displaying recombination breakpoints in the QTL region QSst.lrdc-3B on linkage group 3B. Boxplots showing mean stem solidness score distribution across environments for respective haplotypes. QTL segments marked by blue color lines completely co-segregate with stem solidness. FIGURE 8 Graphical genotypes of wheat haplotypes (H) of doubled haploid lines (DHLs), hollow (Sadash and ‘AAC Innova’)- and solid (P2711)-stem parents, and solid-stem check cultivars (Lillian and ‘Golden Ball’) displaying recombination breakpoints in the QTL region QSst.lrdc-3B on linkage group 3B. Boxplots showing mean stem solidness score distribution across environments for respective haplotypes. QTL segments marked by blue color lines completely co-segregate with stem solidness. FIGURE 8 Graphical genotypes of wheat haplotypes (H) of doubled haploid lines (DHLs), hollow (Sadash and ‘AAC Innova’)- and solid (P2711)-stem parents, and solid-stem check cultivars (Lillian and ‘Golden Ball’) displaying recombination breakpoints in the QTL region QSst.lrdc-3B on linkage group 3B. Boxplots showing mean stem solidness score distribution across environments for respective haplotypes. QTL segments marked by blue color lines completely co-segregate with stem solidness. FIGURE 8 Graphical genotypes of wheat haplotypes (H) of doubled haploid lines (DHLs), hollow (Sadash and ‘AAC Innova’)- and solid (P2711)-stem parents, and solid-stem check cultivars (Lillian and ‘Golden Ball’) displaying recombination breakpoints in the QTL region QSst.lrdc-3B on linkage group 3B. Boxplots showing mean stem solidness score distribution across environments for respective haplotypes. QTL segments marked by blue color lines completely co-segregate with stem solidness. Frontiers in Plant Science 16 frontiersin.org frontiersin.org 10.3389/fpls.2022.1035620 Dhariwal et al. QSst.lrdc-7B) and ‘AAC Innova/P2711’ population (except QSst.lrdc-1B and QSst.lrdc-2A.1) (Table 1). Conversely, a few QTLs such as QSst.lrdc-2D, QSst.lrdc-4A and QSst.lrdc-7B from the ‘AAC Innova/P2711’ population were detected just for internodes but not for whole stem rating (Table 1). Interestingly, most minor QTLs were detected in upper or middle internodes (except QSst.lrdc-5A.2 of ‘Sadash/P2711’ population and QSst.lrdc-2A.2 and QSst.lrdc-5B of ‘AAC Innova/P2711’ population) while major QTLs were detected in all internodes rated in this study (Table 1). Minor loci were largely detected in upper or middle internodes The lower stem internodes in wheat cultivars are generally ﬁlled to the greatest extent (Pluta et al., 2021) and WSS females oviposit eggs between the 2nd to 4th internode (Beres et al., 2011b). Our results indicate that although the effect of these minor QTLs on stem solidness is small and unstable, these can be useful solid stem resources to be synergistically utilized with major loci to enhance the solid stem expression in middle and upper internodes as most of the common wheat lines generally exhibit solidness in lower internodes. QSst.lrdc.3B and QSst.lrdc-3D of ‘Sadash/P2711’ population were analyzed to assess their effectiveness as a single QTL or QTL combination. Similarly, whole stem solidness data of all individual environments and genotyping data of the linked markers IWA1756 and IWB52401 of all the DH lines for major QTLs, QSst.lrdc.3B and QSst.lrdc-3D of ‘AAC Innova/P2711’ population were also analyzed. Solid stem data of DH lines having the same genotypic proﬁle for each group of markers were bulked and plotted as box plot distribution. Based on the QTL proﬁle, the recombinant DH lines in both populations were characterized into four different genotypic classes (Figures 9, 10), irrespective of the solid stem alleles at minor or undetected loci. We observed that while individually, QSst.lrdc-3B contributed maximum stem solidness in both populations, QSst.lrdc-3D either conferred slightly less solidness (in the ‘Sadash/P2711’ population) or almost the same level (statistically insigniﬁcant) of solidness (in ‘AAC Innova/P2711’ population) (Figures 9, 10). Thus, both major loci, QSst.lrdc.3B and QSst.lrdc-3D, were almost equally effective for pith development but together these loci produced a highly solid stem phenotype in both individual wheat populations (Figures 9, 10). QSst.lrdc.3B and QSst.lrdc-3D of ‘Sadash/P2711’ population were analyzed to assess their effectiveness as a single QTL or QTL combination. Similarly, whole stem solidness data of all individual environments and genotyping data of the linked markers IWA1756 and IWB52401 of all the DH lines for major QTLs, QSst.lrdc.3B and QSst.lrdc-3D of ‘AAC Innova/P2711’ population were also analyzed. Solid stem data of DH lines having the same genotypic proﬁle for each group of markers were bulked and plotted as box plot distribution. Based on the QTL proﬁle, the recombinant DH lines in both populations were characterized into four different genotypic classes (Figures 9, 10), irrespective of the solid stem alleles at minor or undetected loci. Synergistic QTL interactions enhance the expression of stem solidness Interestingly, solid stem data distribution showed that some DH lines carrying a single major gene sometimes had less solid stem than the intermediate phenotype (Figures 9, 10). Conversely, some non-major gene carriers conferred stem-solidness that exceeded the intermediate phenotype (Figures 9, 10). These results indicate the involvement of epistatic interactions in both populations. The Individual environment whole stem solidness data and genotyping data of all the DH lines for the linked markers IWB11701 and TG3069, respectively, for major QTLs, FIGURE 9 Violin plot and box plot distribution of stem solidness score in doubled haploid lines carrying (i) minor QTL(s)/none, (ii) major QTL QSst.lrdc-3D, (iii) major QTL QSst.lrdc-3B, and (iv) major QTL combination QSst.lrdc-3B and QSst.lrdc-3D in ‘Sadash/P2711’ population. The violin plots (transparent) and different color-ﬁlled circles represent the data distribution. Quartiles and medians are represented by boxes and continuous lines (gray color), respectively. The mean is shown by dark red color dots. Whiskers extend to the farthest points that are not outliers. Signiﬁcant differences are shown by bar and p values. FIGURE 9 Violin plot and box plot distribution of stem solidness score in doubled haploid lines carrying (i) minor QTL(s)/none, (ii) major QTL QSst.lrdc-3D, (iii) major QTL QSst.lrdc-3B, and (iv) major QTL combination QSst.lrdc-3B and QSst.lrdc-3D in ‘Sadash/P2711’ population. The violin plots (transparent) and different color-ﬁlled circles represent the data distribution. Quartiles and medians are represented by boxes and continuous lines (gray color), respectively. The mean is shown by dark red color dots. Whiskers extend to the farthest points that are not outliers. Signiﬁcant differences are shown by bar and p values. 17 Frontiers in Plant Science frontiersin.org frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 FIGURE 10 Violin plot and box plot distribution of stem solidness score in doubled haploid lines carrying (i) minor QTL(s)/none, (ii) major QTL QSst.lrdc-3D, (iii) major QTL QSst.lrdc-3B, and (iv) major QTL combination QSst.lrdc-3B and QSst.lrdc-3D in ‘AAC Innova/P2711’ population. The violin plots (transparent) and different color-ﬁlled circles represent the data distribution. Quartiles and medians are represented by boxes and continuous lines (gray color), respectively. The mean is shown by dark red color dots. Whiskers extend to the farthest points that are not outliers. Signiﬁcant differences are shown by bar and p values. Synergistic QTL interactions enhance the expression of stem solidness phenotypic characteristics of a QTL may differ in diverse genetic backgrounds due to epistatic interaction with another gene (Dhariwal et al., 2020). This means that an apparently ‘favorable’ QTL allele at a locus may exhibit an ‘unfavorable’ or ‘unexpected’ effect or even a ‘neutral’ phenotype when introgressed to a new genetic background (Kumar et al., 2019). These interactions not only complicate the phenotypic effect but also affects the rate of genetic gain (Kumar et al., 2019), thus, an understanding of epistatic loci is vital for breeding wheat for different traits. Therefore, we further investigated the possibility of synergistic interactions between all loci. We observed that ﬁve of the identiﬁed main effect QTLs (QSst.lrdc-2A, QSst.lrdc-3B, QSst.lrdc-3D, QSst.lrdc-5A.2 and QSst.lrdc-5D) were involved in following 5 digenic epistatic interaction: QSst.lrdc-2A - QSst.lrdc-3B, QSst.lrdc-3D - QSst.lrdc- 5A.2 and QSst.lrdc-3B - QSst.lrdc-3D in ‘Sadash/P2711’ population, and QSst.lrdc-3B - QSst.lrdc-3D and QSst.lrdc-3D - QSst.lrdc-5D in ‘AAC Innova/P2711’ population (Supplementary Table S7). These epistatic interactions conferred an additional increase in pith development in both populations (Supplementary Table S7) conﬁrming their role in the aforementioned phenotypic differences that were observed in the same QTL proﬁle genotypes. Particularly, the digenic epistatic interactions QSst.lrdc-3B - QSst.lrdc-3D and QSst.lrdc-3D - QSst.lrdc-5D were of high interest, which produced a similar effect as that of individual minor loci (Table 1 and Supplementary Table S7). Epistatic interaction of minor loci has also been previously shown to synergistically increase the expression of 3B major locus in Synergistic QTL interactions enhance the expression of stem solidness FIGURE 10 Violin plot and box plot distribution of stem solidness score in doubled haploid lines carrying (i) minor QTL(s)/none, (ii) major QTL QSst.lrdc-3D, (iii) major QTL QSst.lrdc-3B, and (iv) major QTL combination QSst.lrdc-3B and QSst.lrdc-3D in ‘AAC Innova/P2711’ population. The violin plots (transparent) and different color-ﬁlled circles represent the data distribution. Quartiles and medians are represented by boxes and continuous lines (gray color), respectively. The mean is shown by dark red color dots. Whiskers extend to the farthest points that are not outliers. Signiﬁcant differences are shown by bar and p values. phenotypic characteristics of a QTL may differ in diverse genetic backgrounds due to epistatic interaction with another gene (Dhariwal et al., 2020). This means that an apparently ‘favorable’ QTL allele at a locus may exhibit an ‘unfavorable’ or ‘unexpected’ effect or even a ‘neutral’ phenotype when introgressed to a new genetic background (Kumar et al., 2019). These interactions not only complicate the phenotypic effect but also affects the rate of genetic gain (Kumar et al., 2019), thus, an understanding of epistatic loci is vital for breeding wheat for different traits. Therefore, we further investigated the possibility of synergistic interactions between all loci. We observed that ﬁve of the identiﬁed main effect QTLs (QSst.lrdc-2A, QSst.lrdc-3B, QSst.lrdc-3D, QSst.lrdc-5A.2 and QSst.lrdc-5D) were involved in following 5 digenic epistatic interaction: QSst.lrdc-2A - QSst.lrdc-3B, QSst.lrdc-3D - QSst.lrdc- 5A.2 and QSst.lrdc-3B - QSst.lrdc-3D in ‘Sadash/P2711’ population, and QSst.lrdc-3B - QSst.lrdc-3D and QSst.lrdc-3D - QSst.lrdc-5D in ‘AAC Innova/P2711’ population (Supplementary Table S7). These epistatic interactions conferred an additional increase in pith development in both populations (Supplementary Table S7) conﬁrming their role in the aforementioned phenotypic differences that were observed in the same QTL proﬁle genotypes. Particularly, the digenic epistatic interactions QSst.lrdc-3B - QSst.lrdc-3D and QSst.lrdc-3D - QSst.lrdc-5D were of high interest, which produced a similar effect as that of individual minor loci (Table 1 and Supplementary Table S7). Epistatic interaction of minor loci has also been previously shown to synergistically increase the expression of 3B major locus in durum (Nilsen et al., 2017) and common wheat (Cook et al., 2004; Nilsen et al., 2017), however, this is the ﬁrst report of synergistic interaction between major loci QSst.lrdc-3B and QSst.lrdc-3D. Considering digenic epistasis interactions while developing a solid stem cultivar can be useful for achieving higher pith expression in some genetic backgrounds. Supplementary material The ﬁnancial support provided by the Western Grain Research Foundation, and Alberta Wheat Commission to the AAFC Lethbridge spring wheat breeding program is gratefully acknowledged. The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/ fpls.2022.1035620/full#supplementary-material Acknowledgments from P2711 to obtain maximum WSS resistance (Holmes, 1977). P2711 can prove as a valuable source for solid stem transfer to common wheat cultivars. The authors sincerely thank Dr. F. Koekemoer from Sensako, South Africa for providing the genetic material and Dr. Robert J. Graf from AAFC Lethbridge for providing intellectual input for conducting this study. We greatly acknowledge the technical support provided by Leslie Bihari, Julie Pepneck, Marissa Guzzi, Kim Ziegler, Kelly Ryan, Mark Virginillo (AAFC Lethbridge) in phenotyping and ﬁeld experiments and Mira Popovic (AAFC Morden) for genotyping using the 90K Inﬁnium iSelect SNP assay. Conclusion Our ﬁndings suggest that the genetic architecture of stem solidness in both DH populations was largely controlled by two major and stable solid stem QTLs (QSst.lrdc-3B and QSst.lrdc- 3D) from solid stem line P2711 along with several minor QTLs, and some epistatic interactions among detected loci. While both major loci were detected in all internodes, minor loci were largely detected in upper and middle internodes. High expression in upper internodes can be helpful in some genetic backgrounds as solid stem genes are generally ineffective in upper internodes, particularly in common wheat cultivars. Moreover, minor loci can have relatively larger effects when deployed together or with major genes and could also be valuable for diversifying the sources of stem solidness in breeding programs. Conversely, the combination of major solid stem QTLs QSst.lrdc-3B and QSst.lrdc-3D, which produced a highly solid stem phenotype in both populations in this study, can be incorporated in modern wheat cultivars Frontiers in Plant Science 18 frontiersin.org Dhariwal et al. 10.3389/fpls.2022.1035620 Conﬂict of interest RD and HR conceived and designed the study. RD conducted all laboratory and ﬁeld experiments, developed methods, performed all analyses, prepared tables and ﬁgures, interpreted the results, performed project administration and supervision, funding acquisition, wrote – the original draft and performed the review and editing. CH genotyped the mapping populations with 90K Inﬁnium iSelect SNP assay, provided resources, and performed the review and editing. HR developed and grew the mapping populations, performed project administration and supervision, funding acquisition, provided resources, and performed the review and editing. All authors read and approved the ﬁnal manuscript. The authors declare that the research was conducted in the absence of any commercial or ﬁnancial relationships that could be construed as a potential conﬂict of interest. Data availability statement The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/ Supplementary Material. Publisher’s note All claims expressed in this article are solely those of the authors and do not necessarily represent those of their afﬁliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. 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https://openalex.org/W4210349784	https://link.springer.com/content/pdf/10.1007/s11116-022-10267-y.pdf	English	null	Intra-destination travel behavior of alpine tourists: a literature review on choice determinants and the survey work	Transportation	2,022	cc-by	20,908	Transportation (2022) 49:1465–1516 https://doi.org/10.1007/s11116-022-10267-y Transportation (2022) 49:1465–1516 https://doi.org/10.1007/s11116-022-10267-y * Bartosz Bursa bartosz.bursa@uibk.ac.at Intra‑destination travel behavior of alpine tourists: a literature review on choice determinants and the survey work Bartosz Bursa1 · Markus Mailer1 · Kay W. Axhausen2 Accepted: 17 January 2022 / Published online: 4 February 2022 © The Author(s) 2022 Accepted: 17 January 2022 / Published online: 4 February 2022 © The Author(s) 2022 Markus Mailer markus.mailer@uibk.ac.at 1 Unit of Intelligent Transport Systems, University of Innsbruck, 6020 Innsbruck, Austria 2 Institute for Transport Planning and Systems (IVT), ETH Zurich, 8093 Zurich, Switzerland Keywords Tourist travel behavior · Intra-destination · Tourist survey · Survey design · Survey work · Data paper Kay W. Axhausen axhausen@ivt.baug.ethz.ch Introduction and motivation Tourism industry accounts for a substantial share of the direct Gross Domestic Prod- uct (GDP) in the Alpine areas of Austria, Switzerland, Italy or France. In the Austrian province of Tyrol, for example, it is responsible for 17.5% of the GDP (Werbung 2019). However, over the past two decades, vacation trends, characteristics and demands of customers who bring in these revenues have changed significantly. While the average length of stay of tourists in Tyrol decreased from 5.1 nights in 2000 to 4.0 nights in 2019, the number of arrivals increased by almost 60% from around 8 million to more than 12 million (Statistics Austria 2020). This demonstrates the rising demand for tour- ist stays in the province of Tyrol (despite no expansion on the supply side, as the number of beds dropped by 7% between 2000 and 2019), which inevitably leads to an increase in tourism-related travel. In Austria, nearly 75% of inbound holiday trips are made by private car (Austrian National Tourist Office 2014). These trips have a substantial influence on traffic conges- tion on alpine roads and generate pollution in environmentally fragile areas. Car travel, due to its dominant role in Austrian tourism, heavily contributes to the sector’s green- house gas emissions, both at the level of a single destination (Unger et al. 2016), as well as for the entire country (Gühnemann et al. 2021). Tourists appear to be well aware of their travel carbon footprint, yet are not willing to change their behavior substantially (Mailer et al. 2019). Furthermore, traffic generated by tourists can even disrupt daily activities and travel patterns of local inhabitants, who experience severe difficulties in traveling due to lack of alternative roads in densely built-up and narrow mountain val- leys. The problem is recognized and present also in other non-urban destinations, but has so far attracted only limited attention of researchers, who addressed it in qualita- tive studies (Dickinson and Robbins 2007, 2008). Local authorities still do not have any quantitative evidence at their disposal. In effect, transport policy measures in tourist regions are often shots in the dark, which, despite considerable expenses (e.g. free pub- lic transportation for tourists), lack proper evaluation and appraisal. Compared to what we know about everyday travel behavior from representative house- hold surveys, census studies and extensive scientific literature, our knowledge of transpor- tation behavior during vacations is very limited. Abstract In the face of a continuous increase in the number of tourists in the Alps, the associated traffic volumes, and the resulting negative externalities, there is an urgent need to design policies capable of managing tourist traffic efficiently and to invest in transport systems and infrastructure wisely, given the limited financial, spatial and environmental resources. Unfortunately, while there is a considerable research interest in long-distance travel and arrival/departure patterns of tourists, research on tourist mobility during the stay at the destination is almost non-existent. This prevents policy-makers from making informed decisions backed by scientific evidence. The paper presents a data collection study that contributes to filing this gap. This ‘data paper’ reports on the motivation, design and administration of a bespoke travel-activity survey of tourists at vacation destinations. It informs other researchers about the encountered difficulties and helps them optimize data collection effort in their future studies. Besides the contribution to survey methodology, the paper reports on a rare and policy-relevant dataset, provides descriptive results, and thereby contributes to evidence-based design of transport policies in tourist regions. With this paper, we also want to inspire and invite researchers to conduct further quantitative and modeling work in this under-studied field. 1 Unit of Intelligent Transport Systems, University of Innsbruck, 6020 Innsbruck, Austria 2 Institute for Transport Planning and Systems (IVT), ETH Zurich, 8093 Zurich, Switzerland 23456789) 1 3 23456789) 1 3 23456789) 1 3 Transportation (2022) 49:1465–1516 1466 1 3 Transportation (2022) 49:1465–1516 Transportation (2022) 49:1465–1516 1467 The paper is comprised of four sections with Sect. Introduction and motivation intro- ducing the topic, describing the motivation and setting the objectives for the research. Sec- tion State of research on travel behavior of tourists reviews the literature on data collec- tion methods and travel decisions, with a particular focus on tourists in vacation setting. Section Survey design covers the survey work. First, the conceptual framework of tourist travel at the destination is presented. Next, a detailed description of survey methodology and design is given, followed by response behavior statistics from a survey conducted in the field. Section Descriptive analysis provides a descriptive analysis of the collected data. Section Conclusion synthesizes and discusses the findings, and points out the limitations. We also provide an outlook on future research and suggest prospective study topics that could either extend the approach or resolve some of its limitations. Definitions As noticed by Arce and Pisarski (2009), there are many future challenges in describing tourists’ mobility that are caused by i.e. data unavailability, different levels of analysis or inconsistencies in definitions. Therefore, several assumptions were made in the paper to avoid ambiguities. We operate with the definitions of tourism and tourist as proposed by United Nations (2010), so as to avoid confusion with traveler, vacationer or holidaymaker (Terrier 2009). All these terms are used in the paper interchangeably though all meaning a tourist. The main restriction this definition of tourist imposes, is that a person should be out of home (place of residence) for at least one night. It can be either a domestic or an inbound (foreign) visitor. The person must not be specifically on vacation, business pur- poses or family visits are also allowed. It cannot be however a seasonal worker. Of interest are all trips and activities performed during the stay (leisure and non-leisure). Introduction and motivation While these studies provide information on specific vacation decisions (destination preferences, spending, etc.), they look at the issue from the perspective of the country of origin, for which respondents’ behavior dur- ing a stay abroad is not relevant. Similarly, data collected by national tourism agencies and international organizations on tourism markets and aggregate statistics on travelers mov- ing between and within countries are not helpful here either. It is, however, rational and legitimate to assume that travel behavior of tourists at the destination is not only different from their everyday behavior at home (Schlemmer et al. 2019), but it is also dissimilar to travel behavior of local residents. Yet, current research in this field is limited and concen- trates merely on international tourism demand and the long-distance component of vaca- tion travel (Christensen and Nielsen 2018; Gerike and Schulz 2018; Janzen et al. 2018). There are virtually no quantitative data documenting tourists’ travel decisions while at the destination, leaving their travel behavior still largely unexplored.i The fundamental goal of current work is to help fill this research gap by developing a scientifically-grounded approach to collecting data on tourist travel behavior at the des- tination and by providing tourism practitioners, transport planners and policy-makers with a first-of-its-kind dataset from a survey conducted in the Austrian Alps. 1 3 Data collection methods In the case of a smartphone app, a communication channel is necessary to make tourists aware of the app. Furthermore, battery consumption and data roaming in the case of foreign visitors have to be considered. Nonetheless, it is a promising approach and deserves further testing in the field. i Another alternative data source are social media platforms. Recent studies approximated tourist mobility patterns from geo-located Twitter data (Chua et al. 2016; Provenzano et al. 2018), Flickr photos (Önder et al. 2016; Yang et al. 2017) or Foursquare check-ins (Vu et al. 2018). However, in less populated areas, relevant Points-of-Interest are underrepre- sented and geo-tagged tweets and photos are scarce, making these methods more suita- ble studying urban tourism (Sobolevsky et al. 2015) or estimating inter-destination tourist flows (Barchiesi et al. 2015). Moreover, even though such data can deliver valuable infor- mation on tourist activity for the destination managers (Orsi and Geneletti 2013), they are of little use to transportation planners since it is impossible to fully reconstruct all trips made. Nevertheless, despite the expansion of big data, traditional surveys appear to be still in use when investigating tourist populations. Big data on their own are not capable of substituting traditional methods as they do not provide sociodemographic information, cannot measure unobserved variables or deliver strong causal evidence (Chen et al. 2016; Mokhtarian 2018). Unfortunately, as opposed to well-established surveys on daily travel behavior (Brög 2009), there is no consensus on the design and methodology of such surveys in the tourism context that could lead to a replicable approach. Also, very few researchers provide details on the survey design and report on the fieldwork when apply- ing travel diaries (Newmark 2014; Thornton et al. 1997; Tschopp et al. 2010). Authors’ experiences confirm many weaknesses of diary-based surveys of tourists that are also known from surveys of daily mobility, i.e. high costs, low response rate and high drop- out rate. Besides, due to high spatiotemporal dynamics of tourists on site, the sampling frame is unknown and it is difficult to approach a representative sample when surveying outdoors. Surveying visitors at their accommodations allows for more control over sam- pling (e.g. indirect sampling through hotels), but it requires a close cooperation with the accommodation providers, which is usually impossible without the support of local Desti- nation Marketing Organizations (DMO). Even so, self-administered questionnaires distrib- uted through tourism establishments prove very ineffective. Data collection methods The technological progress in recent years has provided academics with new opportuni- ties for measuring mobility by utilizing passively collected big data. Apart from transport researchers also tourism researchers applied tracking technologies in a number of studies (Shoval et al. 2014; Shoval and Ahas 2016). However, these deal with research questions relevant for tourism marketing or tourism geography, but not with the transportation- related aspects of tourist travel, such as traffic generated at destinations or transport mode choice. Mobile positioning data have been widely utilized by tourism researchers in the last decade (Ahas et al. 2008; Zhao et al. 2018). Yet, they proved useful only in applications limited to long-distance travel demand and tourism statistics. In transportation, decisions strongly depend on characteristics of decision-makers (Lu and Pas 1999) and mobile posi- tioning data, for technical and ethical reasons, are lacking this information—only pure location data with time stamps are available. Unfortunately, the density of GSM transceiver stations in mountain regions is typically insufficient for high-resolution analysis at the des- tination level. In addition, in Alpine regions, cross-border trips are very common, resulting in frequent changes of network provider. Thus, only parts of these trips will appear in the dataset obtained from a national provider. 3 3 Transportation (2022) 49:1465–1516 1468 GPS tracking can deliver very fine-grained data on tourist mobility allowing analyses of specific activities or monitoring visitors to facilities, parks and venues (Li et al. 2019). If complemented with additional questionnaires, GPS tracking can serve as a superior alternative to traditional travel surveys among tourists. Currently, mobile phones appear to be used more often in research than independent GPS trackers, since smartphone apps allow for correcting and annotating trips by the user and answering supplementary ques- tions (Prelipcean et al. 2018). Although the first studies reported on failed attempts of GPS tracking with mobile phones (McKercher and Lau 2009), the success rate has increased over the last few years. So far, the most complete and successful approach that combines an annotated travel diary and GPS tracking in a smartphone app for tourist tracking was devel- oped by Hardy et al. (2017), who distributed 240 smartphones with a preinstalled tracking app among visitors to Tasmania. However, besides high costs of such studies, there are also practical and organizational burdens. Tourists cannot be contacted before arrival to arrange handing over the GPS units. Destination choice on vacation Provided that the decision to travel has been taken, decisions must be made about the nature of the trip. The three fundamental dimensions of travel decision-making are destina- tion choice, transport mode choice and route choice. In the context of holiday travel, Bieger and Laesser (2013) distinguish between inter-destination mobility (long-distance travel from home to a destination) and intra-destination mobility (trips made in order to perform activities within the destination area). The choice of vacation destination has always been of interest to researchers from tour- ism marketing and management (Decrop 2006). Understanding these decisions is crucial for tourism-dependent destinations to promote their assets, attract more guests, and as a result, generate more revenue. Therefore, the inter-destination travel patterns have been widely investigated both in theoretical works (Rugg 1973; Sirakaya et al. 1996; Sirakaya- Turk and Woodside 2005; Woodside and Lysonski 1989) and in numerous case studies (Armstrong and Mok 1995; Eymann and Ronning 1997; LaMondia et al. 2010; van Nos- trand et al. 2013). At the same time, the research on the intra-destination movements, i.e. travel within the destination, is relatively limited. As McKercher and Zoltan (2014) argue, the reasons for that are threefold and pertain to the low accuracy of the geolocation data, insufficient resolution of travel-activity data collected from tourists, and lack of a theoreti- cal framework. Only recently, there has been more attention paid to local travel behavior thanks to the use of GPS (Global Positioning System) traces from mobile devices (Shoval et al. 2014; Thimm and Seepold 2016) and GIS (Geographic Information Systems) tech- niques (Lau and McKercher 2006). q Yet, many of the existing studies are descriptive and focus on visualizing geographical and temporal dimensions of tourist movements or drawing conclusions on itinerary types and frequency of visits (McKercher et al. 2019; Wu and Carson 2008). Lew and McK- ercher (2006), in the probably first theoretical work on tourist intra-destination travel, pro- vide an extensive breakdown of factors1 impacting intra-destination movements of tourists, ranging from tourist time budget to personal characteristics to place knowledge. g g g p p g Works utilizing mathematical models are much less prevalent so far. However, the topic is slowly acquiring attention of researchers who start applying discrete choice models to quan- tify travel behavior of tourists and embed them into models. Data collection methods It is therefore postulated that only fully-assisted interviews can guarantee good quality results. Moreover, although travel diary data is detailed enough to model destination and mode choice, it is usually insuf- ficient to investigate route choice. Many of the above issues were discussed by Thornton et al. back in 1997 and are still valid today. 1 3 Transportation (2022) 49:1465–1516 1469 1 Many of these factors are used in the design of the survey instrument (Sect. Survey instrument). Destination choice on vacation A relatively large study on tour- ist local travel (over 2000 face-to-face interviews in 29 tourism destinations) was conducted in three regions in Japan by Wu et al. (2011). Applying a latent class modeling framework, they revealed that, except travel time and distance, attractiveness of a destination (measured by number of attractions and number of visitors) is the main factor influencing destination choice, whereas sociodemographic variables (gender, age, marital status) are decisive for the travel party choice. Researchers have also started exploiting GPS data for model building. For instance, Hardy and Aryal (2020) employed neural networks to analyze GPS tracks of tourist movements in a national park in Australia. Based on survey data and GPS tracks, Li et al. (2019) built models of destination choice of tourist visitors to Gulangyu region in China. They observed that tourists who purchased a joint ticket that includes several attractions tend to travel to zones where these attractions are located. Tourists also avoid areas where they have already been to, and areas with poor signage. As far as the intra-destination mobility within the Alpine regions is concerned, Zoltan and McKercher (2014) analyzed visitors’ behavior in the Swiss canton of Ticino based on destination card use. Their findings reveal that tourist 1 3 3 3 Transportation (2022) 49:1465–1516 1470 movement patterns are defined largely by the spatial dimension rather than through activity- based segmentation. Nevertheless, none of the aforementioned studies distinguishes between movements that are part of tourist activities (e.g. making a hiking trip) and movements to activities (e.g. driving to a zoo), which are of greater importance for transport planning because they cause congestion on roads and crowding in public transportation. Compared to daily travel, which is largely shaped by trips to primary activities (e.g. work, school) with stable locations that do not change at short notice, the choice set of available des- tinations during a vacation stay can be a more complex issue. Unlike residents, visitors do not have equal knowledge about the area and may or may not be aware of some of the alternatives (cf. the choice set formation process by Decrop (2010)) depending on whether they have been to the area before, or whether they have researched the available options in advance. Destination choice on vacation Moreo- ver, they usually have no fixed points governing their movement patterns (other than lodging), while residents are tied to the location where they work or take their children to school, which imposes constraints on their choice set. Due to the short nature of the stay, the visitors’ choice set can be dynamic and change quickly over time (Crompton 1992), making it even more dif- ficult to recognize it in the models. It can be also be driven by habits, attachments or routine (Björk and Jansson 2008), which contradicts the assumption of tourist’s absolute rationality and optimization character of the decision process. For instance, a returning tourist might not consider a new alternative on-site (e.g., a restaurant) since they are used to the one that they have been visiting for the last few years. Besides tourism, there have also been applications of choice models to leisure trips of domestic populations. For instance, Simma et al. (2002) analyze the destination choice for lei- sure activities of Swiss residents within Switzerland. Similarly, Juschten and Hössinger (2021) examined the joint choice of destination and transport mode among the Viennese popula- tion traveling on summer vacation within Austria. Bhat et al. (2016) apply the Multiple Dis- crete–Continuous Probit model to study the leisure destination choice of domestic tourists in New Zealand. To the author’s knowledge, by far the most comprehensive study dealing with leisure and tourism destination choice specifically in the Alpine regions was conducted for Switzerland by Tschopp et al. (2010). Although the objectives and spatial area of their work are similar to the ones defined in current research, they concentrate merely on the arrival/ departure trips to/from the final destinations for both leisure and tourism purposes. Moreover, their destination choice model for holiday trips is limited only to the winter season (skiing activities) and to the trips of Swiss citizens. An example of a more locally and less state-wide focused study is the one by LaMondia and Bhat (2013), who applied the Multivariate Binary Probit model to study the visitors’ leisure travel behavior in Northwest Canada. Scarpa et al. Destination choice on vacation (2008) analyzed the destination choice of members of the Italian Alpine Club (CAI) for one- day outdoor trips in the Alps and discovered that, except travel cost, also difficulty of hiking trails and number of mountain huts influence the decisions, while Scarpa and Thiene (2004) concentrated only on climbers and mountaineers and found travel cost, severity of the environ- ment and number of alpine shelters to be influential factors. Mode choice on vacation Although transport mode choice is relatively well represented in tourism literature, studies using discrete choice methods are very scarce. Much research with discrete choice has been done in the fields of tourism demand (Morley 2012), significantly less in long-haul tour- ism destination choice (LaMondia et al. 2010) and very little in modeling transport mode choice (Thrane 2015). 1 3 1 3 Transportation (2022) 49:1465–1516 1471 Transport mode choice is strongly dependent on the destination choice and hence they should be considered and modeled jointly, which has been repeatedly demanded in the lit- erature (LaMondia et al. 2010; Masiero and Zoltan 2013). Clearly, the decision about trans- port mode choice for local trips within the vacation region depends strongly on the initial decision about transport mode for long-distance trip to the destination (Bieland et al. 2017; Gross and Grimm 2018). However, this relationship also works the other way around—the choice of transport mode for travel to the destination can be driven by factors related to on-site mobility. Visitors decide to travel to Alpine regions by car for fear of insufficient mobility at their destination and inflexibility of public transportation (Bursa and Mailer 2018). In such a case, private car provides a high degree of independence and usually ensures the most effective utilization of time. Luggage transport is another factor deterring tourists from choosing transport mode other than car for their vacation trip (Böhler et al. 2006). As far as the intra-destination mobility of tourists is concerned, a broad description of factors is included in Le-Klähn and Hall (2013). They found out that lack of information and personal preferences are the most common explanations for not using transit services in rural tourism sites. Additionally, not every single tourist spot in rural regions is acces- sible by public transport, which discourages the exploration-focused tourists from relying only on public transport services on-site. In urban areas on the other hand, tourists value the ease of use, efficiency and personal safety when choosing public transport, and parking facilities when driving private car, as Thompson and Schofield (2007) point out. Dickinson and Robbins (2008) also narrowed their research to rural destinations. In addition to gen- eral convenience and need to carry equipment as the main reasons for choosing private car, they also highlight a strong car attachment of some visitors who do not even consider alter- natives no matter their availability, price, or other attributes. Gross and Grimm, in their review paper (2018), synthesized outcomes of many existing studies and found that, above all, sociodemographic factors, transport mode for travel to the destination, travel duration and travel cost, and type of vacation (organized or individual travel) play a role in transport mode choice at the destination. The potential influence of weather is discussed separately in Sect. Route choice on vacation Route choice is another element of the set of decisions that have to be made when plan- ning a trip. The majority of studies on route choice behavior to date have concentrated on dense urban networks. This is understandable because these are the most challenging envi- ronments—urban networks are large, multimodal and the route choice plays a significant role in traffic management. In contrast, research on route choice in non-urban areas is very scarce. Tourism researchers address the topic from the perspective of destination manage- ment and roadside tourism facilities (Denstadli and Jacobsen 2011), which is unusable for transport modeling purposes. However, they provide some interesting observations about how tourists differ in their route choice behavior from local residents, which should be con- sidered when developing models of tourist route choice. p g Lew and McKercher (2006) have raised the issue of tourists not possessing full knowl- edge about the transport system in the region they visit. They highlight the influence of topography on transport networks in mountainous regions, which makes the whole route choice process unlike in flat areas (shortest/fastest route). They also mention the factor of picturesqueness of routes that often prevails over travel time or distance when choosing a route to the destination or moving around within the destination. This is in line with Jacobsen (1996) who discovered that the views and landscape experience are cherished by motor tourists surveyed in Norway. The component of visual attractiveness of a route plays a particularly important role on discretionary (i.e. non-work) trips, according to Ben-Akiva et al. (1984), who found that the disutility of travel time on non-scenic roads is about five times the disutility of travel time on scenic roads. Problematic is however how to define picturesqueness and how to quantify the scenic attributes of a route. Alivand et al. (2015) developed a very promising approach capable of computing scenery-related attributes ranging from road curviness to the viewshed from the road elements using data from different sources and providers e.g. volunteered geographic information (VGI), digi- tal terrain model (DTM), TomTom, Panoramio geo-tagged photos, Google Earth, census data etc. They found that an increased presence of water bodies, mountains, forests and parks along a route positively contributes to the probability of choosing it as a scenic route, whilst urban areas along the route decrease this probability. 1 3 2.6.if Within the alpine setting, specific factors affecting transport mode choice for the long- distance travel as well as for trips at the destination have been investigated by Rüger and Mailer (2020) and Bieger and Laesser (2013), who revealed that the family/group size and transport of sport luggage (e.g., skiing equipment, mountain bike) is dominant in the deci- sion process, particularly in the winter season. Masiero and Zoltan (2013) applied a probit model for the mode choice of tourists in the Swiss canton of Ticino and observed, among other things, that domestic tourists and returning visitors (i.e. tourists who have been to the region before) are more likely to use public transportation, whereas older tourists and male tourists are more inclined to use private cars. The work by Pettebone et al. (2011) provides insights into mode choice at the destination from an American perspective. They found that visitors to the Rocky Mountain National Park are willing to switch from private car to shut- tle bus if it enhances their chances of being in the park with fewer other people. Unfortunately, none of the existing studies analyzed the importance of travel time and travel cost for the transport mode choice of tourists traveling within the destination in a quantitative way, which is a distinct gap in the research, making it impossible to apply a monetary measure to improvements or deteriorations in attributes of the available modes (e.g. higher transit frequency or longer travel time). 3 3 1 Transportation (2022) 49:1465–1516 1472 Impact of weather Typical activities performed by tourists in mountain regions, e.g. hiking, climbing, cycling or skiing, are obviously weather-dependent. It can be argued that if the partici- pation in an activity is weather-dependent, then the choice of location where the activ- ity is conducted and the choice of transport mode used to reach that location may be affected by weather too. It is therefore worth investigating how and to what extent tour- ists at destinations adapt their travel behavior to (unfavorable) weather conditions and whether they react differently than at home. This section provides an overview of what is already known in terms of weather effects on various facets of transportation and tourism. In the context of everyday travel, there is abundant evidence that precipitation cor- relates positively with congestion and accident frequency (Andrey et al. 2003; Golob and Recker 2003). It also negatively affects the performance of transportation networks and traffic flow and thus travel time and travel time reliability (Maze et al. 2006), yield- ing larger effects of snowfall than of rainfall (Hranac et al. 2006). But, as Koetse and Rietveld (2009) conclude in their review paper, the average effects of weather on traf- fic volume, daily travel and commute patterns are of low magnitude and compensate each other in the long run (more leisure trips thanks to higher temperatures; less leisure trips due to longer heavy rains). There is, however, more clear evidence at the level of specific transport modes and instantaneous response of travelers to adverse weather. In particular, the use frequency of active modes—cycling and walking—decreases sig- nificantly in the presence of rain, very low or very high temperatures and strong wind (Sabir 2011; Saneinejad et al. 2012). In the case of cycling, the effect is remarkably large for leisure trips, while bicycle commuters are more weather resistant (Heinen et al. 2010; Liu et al. 2015). Compared to cycling, walking starts to become weather-sensitive in case of a very sharp temperature drop or very intense rain. Studies on weather effects on the choice of private car and public transportation provide weaker outcomes than for active modes. In urban environments, adverse weather interacts strongly with other attributes such as traffic congestion, crowding and punctuality of transit, and the effects differ between cities, days of the week and population segments (Anta et al. 2016). Route choice on vacation The common use of the built-in and external GPS navigation devices among tourists should not be neglected. In the context of car use, it is supposed to lead to tourists some- times having even better knowledge about traffic conditions than local residents, who rely on their habits, common sense and heuristics (Prato et al. 2012). This, however, does not (yet) apply to the knowledge about parking facilities at tourist attractions. Yet, routing decisions of travelers in mountain regions have not been analyzed by schol- ars so far. Presumably because alpine network systems provide limited routing alternatives. However, the valuation of the route’s visual component as well as its windingness, steep- ness and the resulting difficulty to drive may be influential on visitors’ other travel choices. In particular, the attractiveness of the route and the corresponding travel experience may lead to the phenomenon of undirected travel “…in which travel is not a byproduct of the activity but itself constitutes the activity” (Mokhtarian et al. 2001; Mokhtarian and Salo- mon 2001). In other words, visitors might opt for car to be able to drive the scenic roads and stop for taking photos or choose train to observe nature from its panoramic windows. That is, they treat the ride itself as part of the vacation experience, not just the unpleasant necessity of relocation from A to B, which is a valid concept in daily commute (Singleton 2020). 1 3 1 Transportation (2022) 49:1465–1516 1473 Impact of weather In general, however, there is a distinct shift from walking and cycling to driving and transit in case of rain or snow (Sabir 2011). The influence of weather on tourism cannot be neglected, since the whole industry relies to a great extent on “good weather” (Day et al. 2013). Heavy rains, heat waves and frequent storms of increasing severity can negatively affect local tourism markets. Also mountain regions have to face challenges and risks induced by climate change such as increasing snow uncertainty in winter and the resulting decline in demand (Elsasser and Bürki 2002; Gössling and Hall 2006; Koenig and Abegg 1997), or extended sum- mer seasons leading to intense traffic on alpine roads and mountain passes (Caval- laro et al. 2017). However, the evidence on if and how tourists’ weather experiences influence their behavior is complex, ambiguous and segment- and region-dependent (Gössling et al. 2012). While, in general, weather is considered one of the strongest des- tination attributes and one of the most powerful motivators for tourism (Kozak 2002), it must not always be a decisive factor. As far as the planning stage is concerned, accord- ing to Pröbstl-Haider et al. (2015), tourists may not perceive weather as a determinative decision component when choosing an alpine summer destination, as other factors (e.g. leisure attractions) often play a more important role. In terms of the effect during the 1 3 3 3 Transportation (2022) 49:1465–1516 1474 stay, both Scott et al. (2008) and Steiger et al. (2016) found the absence of rain to be particularly important for visitors to mountain areas and their overall satisfaction, with a caveat that there are large differences between age groups, nationalities or first-time and returning tourists. However, satisfaction levels might not necessarily be reflected in the real behavior, and the nature of the visit as well as the attraction mix must be con- sidered. McKercher et al. (2014) analyzed the GPS tracks of visitors to Hong Kong and found that urban tourists are rather indifferent to weather, in particular if they are stay- ing only for a short period or on a tight budget. Similar resilience to weather, increasing with the length of stay, was found among campers in Canada (Hewer et al. 2017). Survey design Researchers conducting studies on daily travel behavior usually have good-quality data- sets at their disposal (see for example Aschauer et al. (2018) and Sammer et al. (2011) for Austria, Axhausen et al. (2000) for Germany, Bundesamt für Statistik (BFS) (2017) for Switzerland, NatCen Social Research (2019) for the UK and Federal Highway Administra- tion (2017) for the US). These studies deal with local populations and serve as a basis for creating policies at regional or national levels. Consequently, government departments and agencies provide funding to ensure sufficient sample size and thus representative results. fi This is not the case in tourist travel, which, despite being responsible for a substan- tial part of transport problems in countries with developed tourism industries (e.g. Austria, Italy, Switzerland), does not receive enough attention from policy-makers.f This study proposes a trade-off and attempts to achieve an acceptable sample size with a limited budget. By asking respondents to report on two days of their stay, fewer people need to be interviewed, which is very time-consuming and costly in the case of tourists. At the same time, limiting the diary length to two days keeps the response burden at a moder- ate level. Impact of weather Also in non-urban settings—as Becken and Wilson (2013) argue in their case study of New Zealand—tourists may have a great dose of understanding and acceptance of inclement weather, in particular if they are aware of and prepared for the unstable and variable weather that often occurs on New Zealand’s islands (which is also the case of the Alps in Europe). They do not search for the most optimal choice, but rather proceed with the plan unless some threshold is exceeded and the weather turns very unappealing. How- ever, whether the same holds for tourists’ short-term transport mode choices or whether the effects of precipitation and temperature on choosing active transport modes are of the same magnitude as known from studies on everyday mobility remains unexplored. 1 3 • Long-distance return trip home. Travel pattern of a tourist The conceptual representation of a full travel pattern of a tourist is given in Fig. 1. In its simplest form (i.e. no “road-trip” with multiple destinations), it is comprised of: • Long-distance trip from home to the destination. • At least one-night-long stay at the destination, during which activities are performed and tours are made. • At least one-night-long stay at the destination, during which activities are performed and tours are made. • Long-distance return trip home. 1 3 Transportation (2022) 49:1465–1516 1475 Fig. 1 Example of a tourist daily schedule with arrival and return trips to and from a tourist destination Fig. 1 Example of a tourist daily schedule with arrival and return trips to and from a tourist destination Fig. 1 Example of a tourist daily schedule with arrival and return trips to and from a tourist destination Survey location Three tourist regions were selected as locations for data collection: the Ötztal valley, the Zillertal valley and the region Hohe Salve. All three are located in the province of Tyrol in Austria (Figs. 2 and 3) and are characterized by extensive tourism industry (Table 1). Survey methods Despite the benefits of automated and semi-automated data collection methods based on GPS (as described in Sect. 2.2 and by Prelipcean et al. (2018)) and obvious drawbacks resulting from using a memory-based approach, the survey was designed as a revealed preference (RP) single cross-sectional survey in two forms: as a PAPI (Paper-and-Pencil 3 Transportation (2022) 49:1465–1516 1476 Fig. 2 Location of the study area on the map of Austria and its neighboring countries. Red-colored rectan- gle is presented in detail in Fig. 3 Fig. 2 Location of the study area on the map of Austria and its neighboring countries. Red-colored rectan- gle is presented in detail in Fig. 3 Fig. 3 Location of the tourist regions Ötztal, Zillertal and Hohe Salve (red dotted areas) in the province of Tyrol (color map) in Austria Fig. 3 Location of the tourist regions Ötztal, Zillertal and Hohe Salve (red dotted areas) in the province of Tyrol (color map) in Austria 1 3 Transportation (2022) 49:1465–1516 Transportation (2022) 49:1465–1516 Transportation (2022) 49:1465–1516 1477 Table 1 Characteristics of the survey regionsa a According to Abteilung Raumordnung und Statistik, Land Tirol, 2019 b Data from winter 2017/18 Ötztal Zillertal Hohe Salve Area [km2] 881 1098 217 Residents 18,277 37,140 15,931 Accommodations 309 467 45 Bedsb 27,865 51,457 5,826 Ski resorts 6 4 1 Ski slopes length [km] 326 535 258 Arrivals – Summer 2019 358,079 666,054 76,766 Overnights—Summer 2019 1,248,163 2,830,628 296,530 Arrivals—Winter 2018/19 618,600 882,405 66,459 Overnights—Winter 2018/19 2,903,563 4,584,125 312,437 Ötztal Zillertal Hohe Salve Interview) survey and CAPI (Computer Assisted Personal Interview) survey conducted with tablet computers.i Interview) survey and CAPI (Computer Assisted Personal Interview) survey conducted with tablet computers.i The choice of these instruments is justified by the characteristics of today’s tourists, who prefer shorter but more frequent stays and booking on short notice, which results in organi- zational difficulty to contact them before the study, equip them with automated devices, instruct and advise during the study and collect the devices before they return home. Fur- thermore, the burden concerning software deployment for semi-automated measurements with smartphones was too large for the study. The initial plan was to conduct the survey exclusively using self-administered PAPI questionnaires distributed in accommodations in the three regions, following the princi- ples proposed by Cambridge Systematics Inc. (1996). This approach would have facilitated the control over the sampling process. Survey methods However, recruiting the hotels to participate in the survey turned out to be a major hurdle. The only successful way to approach the hotel owners about the project was through the local DMO. The DMO employees knew which hoteliers in the area could be potentially willing to cooperate and were capable of convinc- ing some of them to participate. Contacting them directly was ineffective and resulted in refusals justified by lack of time or human resources and concerns about disturbing guests during their vacation time (cf. Tschopp et al. 2010). All in all, however, the response rate proved extremely low (see Sect. 3.5) and hence the survey method was changed to assisted PAPI and CAPI interviews conducted on-site in highly frequented locations spread over the valleys: • Ski lodges, bars and restaurants. • Ski lodges, bars and restaurants. • Local hot-spots such as amusement parks, wellness and spa centers, hot springs. • Local hot-spots such as amusement parks, wellness and spa centers, hot spring • Recreation facilities such as lakes, parks and playgrounds. • Sport facilities such as mountain bike trails, hiking paths. This change resulted in a loss of control over the sampling—a pure convenience sam- pling was now used. It inevitably implies that the results from the sample cannot be easily generalized to the entire study population (Lavrakas 2008; Sirakaya-Turk et al. 2017). A team of trained interviewers conducted interviews on selected days during the winter and summer peak season at various tourist sites. Both fully and partially assisted 3 Transportation (2022) 49:1465–1516 1478 interviewing methods were tested. In the latter method, the interviewers assisted more than one person at a time and switched to a fully assisted personal interview only when they noticed difficulties or someone giving low-quality answers.f fi In the on-site survey, incentives proved ineffective, which to some extent contradicts what is known from the literature (Simmons and Wilmot 2004; Singer and Ye 2012; Tooley 1996). However, studies on the impact of incentives have so far concentrated only on household surveys. Both monetary (5 EUR banknotes) and non-monetary (promotional items) incentives were tested. We could observe that, although incen- tives convinced some negatively oriented guests to fill out the questionnaire (which they presumably would not have done otherwise), the quality of their answers was low (blank fields, inadequate answers). Survey methods On the other hand, among the positively oriented tourists, the motivation to fill out the questionnaire and the quality of their answers was not affected by the incentives, as they were willing participate anyway.i f Regarding the quality of responses, despite the attached instructions for filling out the questionnaire as well as an example of a filled diary, respondents struggled to dis- tinguish between trips and activities. Due to a high response burden, dropouts were not uncommon while filling out the questionnaire. i We argue that only a fully assisted interview and completing the questionnaire in the constant presence of the interviewer can guarantee good-quality results. Tourists approached in local tourist hotspots such as ski lodges must fill out the questionnaire in limited time (lunch break) and space (small tables) and under generally inconvenient conditions (children interrupting, wet clothes). This is a completely different environ- ment compared to household surveys, where respondents can choose a suitable place, moment and take their time (e.g., to read the instructions). Additionally, various inter- action techniques had to be used so as not to deter guests from completing the lengthy questionnaire, such as approaching only selectively chosen tables or approaching chil- dren first, who then, if they find it entertaining, convince parents to participate. Survey instrument Three fundamental parts constitute the survey instrument: • Personal questions. • Personal questions. • Questions about the stay and travel to the destination. • Questions about the stay and travel to the destination. 2 The phenomenon of positive vacation effect on travelers’ happiness and well-being is widely researched and its existence is confirmed (see for example Gilbert and Abdullah (2004) and Sirgy et al. (2011)). There is, however, no consensus on its dynamics over the course of the vacation stay. Nor is there any work so far that studied this phenomenon in the context of perception of travel time or travel cost. Personal questions According to Crawford et al. (1991) and Godbey et al. (2010), participation in leisure activities is subject to intrapersonal, interpersonal and structural constraints. Therefore, in this section, data on factors constituting these constraints were collected using variables such as gender, age, nationality, education/employment status, age and number of children, health/fitness level and car availability. i The selected tourist regions of Ötztal, Zillertal and Hohe Salve are very sport-oriented, both in winter and in summer. While information on sport activity of tourists during their stay is collected through the activity diary, our questionnaire also collects data on guests’ physical activity at home. It is argued that frequency of sport activities in daily life influ- ences sport-related behavior on vacation (De Knop 2007). • Activity diary. • Activity diary. In addition to these elements, the questionnaire also contained text informing the respondent about purpose of the research, research team, processing of personal data, and estimated time for completing the survey. Further, the paper questionnaire con- tained instructions for answering the questionnaire and an example of a completed diary. Figures 17 to 20 in the appendix present the physical design of the paper ques- tionnaires used in the PAPI survey. The CAPI forms were implemented in the SoSci Survey on-line system (Leiner 2020) and have an identical content. The survey instru- ment was prepared in both English and German. The following sections describe the content of the questionnaire and provide reasoning for the selection of questions. 1 3 Transportation (2022) 49:1465–1516 1479 Activity diary The travel-activity diary operates on a selection of terms proposed by Axhausen (2008), that is, trip, tour and activity. An activity-oriented approach was applied for the design of the diary, since it is the activities, not trips that are of greater importance and interest for people during vacation, and hence should prove more effective for the respondents to recall their movements. All travel data was collected at the resolution of trips. Stages (trip legs) were ignored in the survey. In the activity diary, the respondents were asked to give information on all the activities that they performed out of their accommodation during two days of their stay. The diary included questions on the exact type and location of the activity, start/end time, as well as expenses, company and the influence of weather on the activity choice. Furthermore, infor- mation about trips made between the activities was collected, including transport mode, cost, company, impact of weather etc. The activities performed at the accommodation were disregarded, since they do not induce any travel in the transport network. The design of the diary draws from the existing well-established examples of household travel surveys (HTS) including the American NHTS (National Household Travel Survey) (Federal Highway Administration 2017), the German Mobidrive (Axhausen et al., 2000) as well as the Austrian Österreich Unterwegs (Sammer et al. 2011) and MAED (Mobility- Activity-Expenditure-Diary) (Aschauer et al. 2018), all based on the trusted New KON- TIV Design (Brög, 2009), and transposes them into the field of tourism. So as to keep the response burden in the PAPI survey low, the activity diary for two days along with per- sonal, situational and preference questions was fit on a single A3 sheet (half-fold). The PAPI diary takes 50% of the questionnaire (two A4 pages) and provides space for 7 activi- ties and 8 trips per day. In the case of the CAPI survey, the on-line questionnaire included exactly the same questions as the paper version. Automated rules controlled the data qual- ity, correctness of variable types and detected missing answers. This, together with the positive effect of the interviewers conducting the CAPI survey, resulted in noticeably better quality of the collected data.i In contrast to typical HTS questionnaires, we decided not to provide any predefined activity types in the questionnaire. Questions about the stay and travel to the destination Lew and McKercher (2006) have classified the factors influencing local travel behavior of tourists into three categories: • The size and expenditure of tourist time budget. • Personal motivations, interests and travel group composition. • Tourist knowledge of the destination. In this section, respondents were asked basic questions about the length of stay, exact place of stay, travel party composition and transport mode used for travel to the destination. By collecting information on the exact arrival and departure dates of visitors, one can ana- lyze the effect of the (fixed) length of stay, as well as the effect of the (varying) moment of the stay, represented by the elapsed fraction of stay. This is driven by two hypotheses. The first one posits that with the vacation days going by, people are becoming more relaxed,2 and hence, may react less negatively to travel time. Their positive attitude might also fol- low a non-linear curve, as proposed by Lin et al. (2014), reaching its peak around the mid- dle of the stay. The second one assumes the opposite—visitors are becoming more stressed and impatient and hence are reacting more negatively, which would be in line with the find- ings of Nawijn et al. (2013). According to LaMondia and Bhat (2013), tourists tend to have a main purpose charac- terizing the long-distance activity component of their holiday trip, which then drives them to choose a particular destination and particular activities. The topic of holiday and lei- sure motivations has been intensively studied by many researchers who developed different measurement scales and items (Beard and Ragheb 1983; Crompton 1979; Iso-Ahola 1984; Ryan and Glendon 1998). Given the restricted space in the questionnaire, eight predefined purpose categories were proposed (see the Appendix). According to Lehto et al. (2004), whether a tourist has visited a destination before or it is their first visit, affects their knowledge about the destination (activities and attrac- tions, local transportation), which ultimately influences their activity and travel choices on- site. The same applies to whether a destination is the main and only place of stay during 1 3 3 3 Transportation (2022) 49:1465–1516 1480 vacation or is it one of many stops. A touring trip implies different on-site behavior than stationary vacation (Lew and McKercher 2006). Questions regarding these two aspects have been incorporated in the survey. Questions about the stay and travel to the destination The information search behavior is considered crucial for tourists’ knowledge about the destination (Bieger and Laesser 2016; Fodness and Murray 1999; Gursoy and McCleary 2004; Klassen 2001). Therefore, two additional questions were asked, aiming to find out how, if at all, visitors inform themselves in advance about the journey to the destination and about the mobility on-site. Survey participation and response burden As long as the choice and wording of survey questions and the definition of the survey area were relatively comprehensive and manageable tasks, the actual implementation of the survey in a tourist region was a much more complex undertaking. The most difficult issues included the choice of the survey method, distribution method for the PAPI questionnaires, arranging meetings with accommodation providers, convincing them to participate in the project, defining incentives and finding a way to reach out to guests in the field and to over- come their participation and response burden. A summary of these efforts is provided in Table 2. For the paper questionnaire with a travel-activity diary for two days, the total response burden was calculated following the methodology by Axhausen et al. (2015). In the mini- mum scenario of only one activity and two trips per day, the response burden is 381, whereas the maximum case of seven activities and eight trips (all boxes filled) results in a response burden of 1309. Employing their regression equation, one would expect a response rate of 8.00–24.35% respectively. Out of the 640 questionnaires (270 in English, 370 in German) distributed in the hotels during the winter season, only 28 were returned, yielding an average response rate of 4.4%. It is below the lower bound of the range estimated using the method by Axhausen et al. (2015). Potential reasons for that are: • Difficulties in recruiting hotels for the study that are truly willing to cooperate. fi • No control over if, how, when and to whom the questionnaires were distributed after they were handed over to the hotel.f • Possibly negative mediating role of the reception staff (not all questionnaires were dis- tributed; guests were not sufficiently encouraged to participate). fi • Questionnaires were lost in several cases. In the face of a very low response rate, the remaining part of the PAPI survey was con- ducted in form of (semi-) assisted interviews where dropouts were no longer noted. There were refusals when interviewers approached potential respondents, but interviewers were not obliged to report this. In the CAPI survey, which was also conducted in the form of an interview as well, all started interviews were completed, and denials were not reported. Activity diary While reliable and validated categories have been developed for daily activities (e.g., work, education, pick-up/drop-off, shopping, leisure), it is very difficult to create categories for all possible leisure activities (although there are some attempts, see Lanzendorf (2002)). Therefore, it is common to ask the respondent for their own detailed description of the performed activity (Axhausen 2015) and classify it afterwards.l With regard to the influence of weather on the activity and mode choice, the respond- ents were asked to indicate whether they chose the activity/mode that they had planned to choose or whether they had to choose another (“plan B”) activity/mode because of the 1 3 Transportation (2022) 49:1465–1516 1481 (unfavorable) weather. This novel approach makes it possible to directly capture the impact of weather on every decision made during the reported day. In combination with historical weather measurement data for the survey dates, it is a very powerful dataset. In the few existing studies (Liu et al. 2016; Termida et al. 2016), only the information on subjective weather perception on a given day was collected. Information about lodging comprises: Information about lodging comprises: • object type (hotel, guesthouse, apartment, camping). • standard (only for hotels, represented by star rating). • price per person per night (in EUR) in summer and winter. • price per room/apartment per night (in EUR) in summer and winter. Complementary datasets • Umhausen, 1035 m a.s.l. • Obergurgl, 1942 m a.s.l. • Mayrhofen, 640 m a.s.l. • Söll, 697 m a.s.l. • Innsbruck, 578 m a.s.l. Information about lodging comprises: • object type (hotel, guesthouse, apartment, camping). • standard (only for hotels, represented by star rating). • price per person per night (in EUR) in summer and winter. • price per room/apartment per night (in EUR) in summer and winter. • Haiming, 669 m a.s.l. • Umhausen, 1035 m a.s.l. • Obergurgl, 1942 m a.s.l. • Mayrhofen, 640 m a.s.l. • Söll, 697 m a.s.l. • Innsbruck, 578 m a.s.l. • Haiming, 669 m a.s.l. • Umhausen, 1035 m a.s.l. • Obergurgl, 1942 m a.s.l. • Mayrhofen, 640 m a.s.l. • Innsbruck, 578 m a.s.l. Complementary datasets The survey results comprise the primary data source used in the paper. These data are com- plemented by following secondary datasets: 1 3 1 3 Transportation (2022) 49:1465–1516 1482 Table 2 Summary of the survey protocol depending on survey region, wave, method and language Season Region Wave Method Incentives Language Conducted PAPI/ CAPI interviews Distributed PAPI questionnaires Returned PAPI questionnaires Summer Ötztal 16–19.7.2019 CAPI Promotional items EN 15 – – DE 139 – – 31.7–2.8.2019 CAPI Promotional items EN 2 – – DE 12 – – 31.7–3.8.2019 CAPI Promotional items EN 4 – – DE 26 – – 20–23.8.2019 CAPI Promotional items EN 17 – – DE 109 – – Zillertal 26–30.7.2019 CAPI Promotional items EN 4 – – DE 44 – – 26.8–4.9.2019 CAPI Promotional items EN 1 – – DE 38 – – Winter Ötztal 12.2018–4.2019 PAPI self– administered No EN – 270 0 DE – 370 28 25–27.12.2018 PAPI Promotional items EN 45 – – DE 41 – – 4–5.1.2019 PAPI 5 EUR banknotes EN 14 – – DE 75 – – 27.2–1.3.2019 PAPI Promotional items EN 12 – – DE 60 – – Zillertal 18–20.2.2019 PAPI Promotional items EN 40 – – DE 77 – – Hohe Salve 8.3.2019 PAPI Promotional items EN 15 – – DE 31 – – Sum 821 640 28 Transportation (2022) 49:1465–1516 1483 • Historical weather data. • Historical weather data. • Geodata from Google Maps API. • Geodata from Google Maps API. • Geodata from Google Maps API. g p • Geodata from the regional transportation model. • Geodata from the regional transportation model. g p • Accommodation data from booking systems of the tourist regions. • Accommodation data from booking systems of the tourist regions. Historical weather data were obtained from the Central Institution for Meteorology and Geodynamics (Zentralanstalt für Meteorologie und Geodynamik, ZAMG) in Austria. The data include measurements of air temperature, precipitation, cloud cover, wind speed and snow depth, and were collected at one-hour resolution from following six monitoring stations: • Haiming, 669 m a.s.l. • Umhausen, 1035 m a.s.l. • Obergurgl, 1942 m a.s.l. • Mayrhofen, 640 m a.s.l. • Söll, 697 m a.s.l. • Innsbruck, 578 m a.s.l. Information about lodging comprises: • object type (hotel, guesthouse, apartm • standard (only for hotels, represented • price per person per night (in EUR) in • price per room/apartment per night (in • Haiming, 669 m a.s.l. • Geodata from Google Maps API. Data cleansing In order not to lose valuable observations where only few items were missing, it was neces- sary to impute the missing data. The multiple imputation method was chosen (van Buuren 2018), as it delivers less biased results than the ad-hoc solutions (e.g. mean imputation) (Andridge and Little 2010). The missForest package for R (Stekhoven and Bühlmann 2012) was used for the following variables: income, age, gender, education, employment, country of residence, nationality, car availability, car use frequency, driver’s license pos- session, main transport mode for travel to the destination, main purpose of stay, number of adults in household, number of children under 6, number of children aged 6 to 17, length of the stay, knowledge about travel options to the destination, knowledge about on-site mobil- ity, sport frequency, time spent on sport,, and hotel-related variables. The results presented in the following sections are based on data after the imputation process. Regardless of the data imputation, following exclusion rules were employed: • The only respondent from a group/family is below 18 (assumed not to be the decision- maker in a family/group). • The only respondent from a group/family is below 18 (assumed not to be the decision- maker in a family/group). R d t’ l f t i t id th t d • Respondent’s place of stay is outside the study area. The answer quality is unacceptable (contradictory answers, misunderstood questions). • The answer quality is unacceptable (contradictory answers, misunderstood questio As a result, out of 849 questionnaires 224 were eliminated (predominantly PAPI ques- tionnaires from winter) and 625 remained (388 in summer and 237 in winter) (Table 3). 3 Transportation (2022) 49:1465–1516 1484 Table 3 Valid responses depending on survey method and season Share of responses Summer Winter Number % Number % Self-administered PAPI 0 0 25 10.5 PAPI 0 0 212 89.5 CAPI 388 100 0 0 Sum per season 388 62.1 237 37.9 Sum total 625 The descriptive results are reported at the level of individual respondents (unless otherwise specified). The descriptive results are reported at the level of individual respondents (unless otherwise specified). Socio‑demographics Table 4 provides a statistical summary (group frequencies and mean values) of the respond- ents’ sociodemographic characteristics from the summer and winter survey periods.f There are notable differences between the seasons. Winter tourists are substantially younger and better educated. They are also more professionally active and possess higher incomes. Their health is better, possibly because they are more physically active. Summer visitors on the other hand are older, less educated, often already retired or working part- time. They have less disposable income, lower car availability and use private cars less often than winter guests. Visitors from Germany dominate in both seasons. Noteworthy, they are followed by Austrian domestic tourists in summer, whilst in winter Dutch tourists come in second place. Figure 4 illustrates correlations between selected sociodemographic variables. The cat- egorical variables with a self-explanatory order of levels, that is, education, employment, income, and health, were transformed into numeric variables, assuming the lowest level equals 1 and all higher levels are equidistant. Stay and travel to the destination Responses to questions concerning the stay and travel to the destination (part 2 of the ques- tionnaire) are summarized in Table 5. Overall, summer stays are longer than winter stays. The high standard deviation sug- gests a wide spread of stay durations in summer. In winter, the length of stay oscillates around 6–7 overnights, which corresponds to a typical holiday week that starts and ends on a Saturday. A winter tourist is accompanied by more household members. Both summer and winter tourists prefer individually organized holidays, which comprise around 85% of all stays. In over 50% cases they have already been to Tyrol before. Winter stays are pre- dominantly stationary, whereas in summer, a 15% share of respondents declare being on a road trip and moving to another place soon. An average winter tourist comes to almost 80% for sport and recreation, while a summer visitor is attracted similarly by sport (37%) and rest and relaxation (30%), followed by social and health purposes. Stay and travel to the destination 1485 Transportation (2022) 49:1465–1516 le 4 Sociodemographic description of the sample iable Value Summer Winter Number % Number % e Mean (SD): 47.7 (15.0) Mean (SD): 39.3 (14.2) 6–17 4 1.0 7 3.0 18–24 26 6.7 44 18.6 25–40 98 25.3 69 29.1 41–64 207 53.4 111 46.8 65 + 53 13.7 6 2.5 nder Female 210 54.1 109 46.0 Male 176 45.9 128 54.0 idence country Germany 238 61.3 142 59.9 Austria 82 21.1 12 5.1 Netherlands 20 5.2 38 16.0 Switzerland 11 2.8 6 2.5 Italy 6 1.6 0 0.0 France 5 1.3 1 0.4 UK 5 1.3 21 8.9 Other 21 5.4 17 7.2 cation Primary level 25 6.4 5 2.1 Secondary level (high school) 125 32.2 36 15.2 A-levels / High school diploma 98 25.3 72 30.4 University degree (Bachelor’s, … 140 36.1 124 52.3 Transportation (2022) 49:1465–1516 1486 1 3 Table 4 (continued) Variable Value Summer Winter Number % Number % Employment Full-time employed 191 49.2 135 57.0 Retired 72 18.6 7 3.0 Part-time employed 54 13.9 13 5.5 Pupil or student 27 7.0 43 18.1 Doing housework, looking after… 21 5.4 8 3.4 Self-employed / own business 21 5.4 28 11.8 Unemployed or looking for a job 2 0.5 0 0.0 Apprentice or trainee 0 0.0 3 1.3 Household size Mean (SD): 2.72 (1.4) Mean (SD): 2.85 (1.28) 3 1 0.3 0 0.0 4 59 15.2 39 16.5 5 151 38.9 66 27.9 6 57 14.7 49 20.7 7 86 22.2 60 25.3 8 27 7.0 21 8.9 > 8 7 1.9 2 0.8 Number of children under 6 in the household 1 334 86.1 0 0 2 40 10.3 214 90.3 3 13 3.4 15 6.3 4 1 0.3 8 3.4 1487 Transportation (2022) 49:1465–1516 le 4 (continued) iable Value Summer Winter Number % Number % mber of children 6–17 in the household 1 274 70.6 152 64.1 2 57 14.7 46 19.4 3 47 12.1 28 11.8 4 8 2.1 11 4.6 5 2 0.5 0 0 nthly net household income in EUR < 1,000 20 5.2 26 11.0 1,000–2,000 53 13.7 16 6.8 2,001–3,000 106 27.3 26 11.0 3,001–4,000 95 24.5 39 16.5 4,001–5,000 35 9.0 41 17.3 5,001–6,000 14 3.6 17 7.2 6,001–7,000 22 5.7 22 9.3 7,001–8,000 8 2.1 12 5.1 8,001–9,000 4 1.0 14 5.9 9,001–10,000 3 0.8 4 1.7 10,001–12,000 3 0.8 3 1.3 12,001–14,000 8 2.1 2 0.8 14,001–16,000 3 0.8 2 0.8 16,001–18,000 10 2.6 3 1.3 18,001–20,000 1 0.3 1 0.4 > 20,000 3 0.8 9 3.8 nual leave (days) Mean (SD): 17.6 (6.3) Mean (SD): 17.7 (6.9) Monthly net household income in EUR Transportation (2022) 49:1465–1516 1488 le 4 (continued) iable Value Summer Winter Number % Number % hts away in the last year I did not go away 8 2.1 2 0.8 1–5 nights 23 5.9 8 3.4 6–10 nights 70 18.0 18 7.6 11–20 nights 144 37.1 54 22.8 21–30 nights 82 21.1 69 29.1 More than 30 nights 61 15.7 86 36.3 ver’s license No 20 5.2 23 9.7 Yes 368 94.8 214 90.3 availability Never 54 13.9 18 7.6 Sometimes 44 11.3 28 11.8 Always 290 74.7 191 80.6 use frequency Less than once a month 26 6.7 8 3.4 1–3 times a month 45 11.6 29 12.2 1–3 days a week 92 23.7 42 17.7 4–6 days a week 64 16.5 37 15.6 Daily 161 41.5 121 51.1 alth status Very bad 2 0.5 1 0.4 Bad 5 1.3 0 0.0 Fair 52 13.4 6 2.5 Good 154 39.7 97 40.9 Very good 175 45.1 133 56.1 1489 Transportation (2022) 49:1465–1516 e 4 (continued) able Value Summer Winter Number % Number % sical limitations Severely limited 6 1.6 2 0.8 Limited but not severely 87 22.4 18 7.6 Not limited at all 295 76.0 217 91.6 rt frequency (days in a week) 0 63 16.2 13 5.5 1 76 19.6 34 14.4 2 88 22.7 65 27.4 3 59 15.2 53 22.4 4 38 9.8 23 9.7 5 35 9.0 26 11.0 6 6 1.6 9 3.8 7 23 5.9 14 5.9 rt time (hours in a week) Mean (SD): 3.70 (3.6) Mean (SD): 4.60 (4.0) 1 3 Limited but not severely 3 1 Transportation (2022) 49:1465–1516 1490 Fig. 4 Correlations of the decision-makers’ characteristics Fig. 4 Correlations of the decision-makers’ characteristics In terms of transport mode used to travel from home to the destination, private car with around 80% share dominates in both seasons. This is followed by air travel in winter (mostly from the UK thanks to a convenient direct connection from London to Innsbruck) and by train in summer. The major reason for choosing a particular trans- port mode is convenience (most direct, accessible and flexible mode) in both seasons. In winter, luggage plays an important role, as do price, journey time and distance. In summer, factors other than convenience are far less important. Figure 5 illustrates the relationship between the chosen transport mode and the factors specified as driving this decision. Visitors in both seasons declare to be generally well informed about the jour- ney options. However, only winter tourists state that they are sufficiently informed about the transportation at the destination.i In terms of accommodation types, summer tourists definitely prefer hotels (65%) to guesthouses (19%), apartments and campsites (Table 6). Winter tourists lean towards hotels (44%), but also choose guesthouses relatively often (35%). Average prices (per person per night) are, even despite a higher share of guesthouse stays in the sample, about 15 EUR higher in winter than in summer, which clearly implies that winter is the more expensive season. Limited but not severely 1 3 1 3 Transportation (2022) 49:1465–1516 1491 p ( ) Table 5 Description of the stay and travel to the destination Variable Value Summer Winter Number % Number % Length of stay Mean (SD): 8.70 (5.1) Mean (SD): 6.25 (1.84) 1–5 95 25.3 68 28.7 6–10 178 45.9 165 69.6 11–15 62 16.0 3 1.3 > 16 50 12.9 1 0.4 Company during the stay Household size Mean (SD): 2.44 (1.4) Mean (SD): 2.77 (1.9) Alone Yes 43 11.1 5 2.1 No 345 88.9 232 97.9 With a spouse Yes 271 69.9 127 53.6 No 117 30.1 110 46.4 Children under 6 0 338 87.1 213 89.9 1 37 9.5 13 5.5 2 12 3.1 8 3.4 > 2 1 0.3 3 1.3 Children 6–17 0 280 72.2 154 65.0 1 53 13.7 46 19.4 2 43 11.1 26 11.0 > 2 12 3.1 11 4.6 Other household members 0 365 94.1 188 79.3 1 12 3.1 11 4.6 2 4 1.0 22 9.3 > 2 7 1.8 16 6.8 ription of the stay and travel to the destination Value Summer Winter Number % Number % y Mean (SD): 8.70 (5.1) Mean (SD): 6.25 (1.84) 1–5 95 25.3 68 28.7 6–10 178 45.9 165 69.6 11–15 62 16.0 3 1.3 > 16 50 12.9 1 0.4 ing the stay Household size Mean (SD): 2.44 (1.4) Mean (SD): 2.77 (1.9) Yes 43 11.1 5 2.1 No 345 88.9 232 97.9 e Yes 271 69.9 127 53.6 No 117 30.1 110 46.4 er 6 0 338 87.1 213 89.9 1 37 9.5 13 5.5 2 12 3.1 8 3.4 > 2 1 0.3 3 1.3 7 0 280 72.2 154 65.0 1 53 13.7 46 19.4 2 43 11.1 26 11.0 > 2 12 3.1 11 4.6 old members 0 365 94.1 188 79.3 1 12 3.1 11 4.6 2 4 1.0 22 9.3 > 2 7 1.8 16 6.8 Table 5 Description of the stay and travel to the destination Transportation (2022) 49:1465–1516 1492 1 3 Table 5 (continued) Variable Value Summer Winter Number % Number % Other known persons 0 302 77.8 133 56.1 1 29 7.5 23 9.7 2 16 4.1 22 9.3 > 2 41 10.6 59 24.9 Type of holiday Individual trip Organized travel (by tour operator, … 327 84.5 197 85.3 60 15.5 34 14.7 Number of previous visits 0 177 46.0 96 40.9 1 50 13.0 28 11.9 2 40 10.4 21 8.9 3–5 51 13.3 36 15.3 6–10 28 7.3 31 13.2 > 10 39 10.1 23 9.8 Main destination Yes 330 85.7 227 97.4 No, I am on a stopover here and will… 55 14.3 6 2.6 Main purpose Business 4 1.0 0 0.0 Culture, heritage, sightseeing 16 4.1 0 0.0 Health, wellness 47 12.1 6 2.5 Rest, relaxation 115 29.6 26 11.0 Shopping, fun, entertainment 2 0.5 5 2.1 Social (time with family, friends) 60 15.5 13 5.5 Sport, recreation 144 37.1 187 78.9 e 5 (continued) iable Value Summer Winter Number % Number % er known persons 0 302 77.8 133 56.1 1 29 7.5 23 9.7 2 16 4.1 22 9.3 > 2 41 10.6 59 24.9 e of holiday Individual trip Organized travel (by tour operator, … 327 84.5 197 85.3 60 15.5 34 14.7 mber of previous visits 0 177 46.0 96 40.9 1 50 13.0 28 11.9 2 40 10.4 21 8.9 3–5 51 13.3 36 15.3 6–10 28 7.3 31 13.2 > 10 39 10.1 23 9.8 n destination Yes 330 85.7 227 97.4 No, I am on a stopover here and will… 55 14.3 6 2.6 n purpose Business 4 1.0 0 0.0 Culture, heritage, sightseeing 16 4.1 0 0.0 Health, wellness 47 12.1 6 2.5 Rest, relaxation 115 29.6 26 11.0 Shopping, fun, entertainment 2 0.5 5 2.1 Social (time with family, friends) 60 15.5 13 5.5 Sport, recreation 144 37.1 187 78.9 1493 Transportation (2022) 49:1465–1516 Table 5 (continued) Variable Value Summer Winter Number % Number % Main transport mode used for travel to the destination Airplane 7 1.8 23 9.7 Coach 18 4.6 10 4.2 Motorcycle as a driver 2 0.5 0 0.0 Private car as a driver 180 46.4 101 42.6 Private car as a passenger 138 35.6 94 39.7 Rented car, car-sharing as a driver 3 0.8 1 0.4 Rented car, car-sharing as a passenger 3 0.8 1 0.4 Train 37 9.5 7 3.0 Reason for choosing this modea No other mode was available 55 14.2 9 3.8 Because of the distance of the journey 50 12.9 72 30.6 Fastest mode 51 13.1 79 33.6 Cheapest mode 54 13.9 70 29.8 Safest mode 7 1.8 16 6.8 Most convenient modeb 236 60.8 120 51.1 Most comfortable mode 59 15.2 56 23.8 Personal mobility constraints 9 2.3 3 1.3 Luggage transport 84 21.7 101 43.0 Weather conditions 7 1.8 5 2.1 Other 46 11.9 5 6.3 Transportation (2022) 49:1465–1516 1494 1 3 Table 5 (continued) Variable Value Summer Winter Number % Number % Information level about travel options to the destination Not informed at all 84 21.7 45 19.0 Slightly informed 30 7.7 13 5.5 Somewhat informed 46 11.9 43 18.1 Well informed 128 33.0 83 35.0 Very well informed 100 25.8 53 22.4 Source of this informationa On websites/mobile app of the region/… 92 23.7 81 34.8 On websites/mobile app of the hotel 18 4.6 26 11.2 On online/mobile map services… 97 25.0 56 24.0 At the travel agency 9 2.3 10 4.3 From travel guidebooks 8 2.1 2 0.9 From friends and relatives 18 4.6 35 15.0 Other 56 14.4 13 5.5 I have not informed myself in advance 203 52.3 84 36.1 Information about the on-site mobility Not informed at all 97 25.0 22 9.3 Slightly informed 46 11.9 19 8.0 Somewhat informed 56 14.4 44 18.6 Well informed 105 27.1 86 36.3 Very well informed 84 21.7 66 27.9 e 5 (continued) able Value Summer Winter Number % Number % rmation level about travel options to the destination Not informed at all 84 21.7 45 19.0 Slightly informed 30 7.7 13 5.5 Somewhat informed 46 11.9 43 18.1 Well informed 128 33.0 83 35.0 Very well informed 100 25.8 53 22.4 rce of this informationa On websites/mobile app of the region/… 92 23.7 81 34.8 On websites/mobile app of the hotel 18 4.6 26 11.2 On online/mobile map services… 97 25.0 56 24.0 At the travel agency 9 2.3 10 4.3 From travel guidebooks 8 2.1 2 0.9 From friends and relatives 18 4.6 35 15.0 Other 56 14.4 13 5.5 I have not informed myself in advance 203 52.3 84 36.1 rmation about the on-site mobility Not informed at all 97 25.0 22 9.3 Slightly informed 46 11.9 19 8.0 Somewhat informed 56 14.4 44 18.6 Well informed 105 27.1 86 36.3 Very well informed 84 21.7 66 27.9 1495 Transportation (2022) 49:1465–1516 Table 5 (continued) Variable Value Summer Winter Number % Number % Source of this informationa On websites/mobile app of the region/… 89 22.9 95 41.0 On websites/mobile app of the hotel 34 8.8 46 19.8 On websites/mobile apps of the local… 35 9.0 16 6.9 On online/mobile map services… 37 9.5 30 12.9 At the travel agency 7 1.8 5 2.2 From travel guidebooks 16 4.1 4 1.7 From friends and relatives 10 2.6 36 15.5 Other 54 13.9 23 9.7 I have not informed myself in advance 219 56.4 53 22.7 a Multiple choice question. Limited but not severely Values indicate percent share of people who chose one of the answers b Convenient mode was defined in the questionnaire as direct, accessible and flexible, whereas comfort pertained to e.g. cleanliness, seats and ventilation. See the items used in question 28 in the questionnaire in the appendix 1 Transportation (2022) 49:1465–1516 1496 Fig. 5 Reasons for choosing particular transport mode for travel to the destination (multiple choice pos- sible) Fig. 5 Reasons for choosing particular transport mode for travel to the destination (multiple choice pos- sible) 1 3 Activities Figure 6 presents locations of the accommodations where the respondents stayed. Figure 7 illustrates activity locations (i.e. trip start points). The area overlaps to a large extent with Fig. 6, which means that activities are performed predominantly within the valley, in the vicinity of the place of stay. Tourists make excursions outside their region relatively rarely. The locations of activities are very concentrated in winter and are close to the main road axis and ski resorts, whereas in summer they are more uniformly distributed across the regions and are more distant from the center of the regions. Visits to places outside the valleys, such as picturesque lakes (Achensee) or cities with tourist attractions (Innsbruck, Schwaz, Kufstein) were reported more fre- quently in summer than in winter. 1 3 1497 Transportation (2022) 49:1465–1516 Table 6 Characteristics of the accommodations reported in the survey a Only for hotels. Not available for apartments, campsites and guesthouses Variable Value Summer Winter Number % Number % Type of accommodation Apartment 42 10.8 49 20.7 Camping 19 4.9 3 1.3 Hotel 252 65.0 103 43.5 Guesthouse 75 19.3 82 34.6 Standard(number of stars)a 2 2 0.8 0 0 3 50 19.8 15 14.6 4 165 65.5 72 69.9 4.5 35 13.9 13 12.6 5 0 0 3 2.9 Price per person per night [EUR] Mean (SD): 68.4 (32.9) Mean (SD): 82.3 (39.3) Table 6 Characteristics of the accommodations reported in the survey a Only for hotels. Not available for apartments, campsites and guesthouses Fig. 6 Locations of the accommodations reported in the survey Fig. 6 Locations of the accommodations reported in the survey 3 There are no fully comparable data on trips rates available for the UK—only for England. Mobility rates The general level of mobility of tourists at the destination can be described by number of trips, kilometers traveled and time spent on travel. The average number of trips per person per day in the sample is 2.5 (Table 7). Compared to the values generated by the residents of Tyrol, tourists appear to be less mobile. The difference becomes even more apparent when compared to mobility rates of residents in municipalities with intensive tourism, who make 3.8 trips per day (Köll and Bader 2011). Similarly, in comparison with mobility lev- els measured in countries where the visitors come from, mobility levels during vacation 3 Transportation (2022) 49:1465–1516 1498 Fig. 7 Location of the activities depending on survey location Fig. 7 Location of the activities depending on survey location are also trending lower (except for the UK3 (NatCen Social Research 2019)). This implies that people are generally less mobile during their out-of-home stays than on a typical day at home. However, one should point out regional variations within these countries and meth- odological differences between the studies before generalizing the results. f In terms of distance traveled, tourist cover substantially fewer kilometers per day on vacation than on an average day at home. However, there are large country-dependent dif- ferences, with Austrians traveling the shortest and Germans the longest distances. In terms of time spent on travel, both the Austrian and foreign tourists achieve significantly lower values during vacation than when at home. Only German visitors stand out, spending on average 75 min per day compared to 80 min in their daily behavior. Transport mode choice of the respondents (2015) g Bundesamt für Statistik (BFS) (2017) h NatCen Social Research (2019) Study population Daily distance [km] Daily travel time [min] No of trips per daya Tourists – total 23.2 (28.8/14.2)b 59 (72 / 39)b 2.5 (2.4/2.7)b Tourists from AT 11.3 22 2.1 Tourists from DE 27.2 75 2.6 Tourists from NL 16.4 38 2.7 Tourists from CH 22.3 45 2.2 Tourists from UK 21.1 26 2.9 Residents in AT (2013/2014)c 36 68 2.6 Residents in AT, Tyrol (2013/2014)c 35 69 2.7 Residents in AT, Tyrol (2011)d – – 4.1 Residents in AT, Tyrol (intensive tourist municipalities) (2011)d – – 3.8 Residents in DE (2018)e 39 80 3.1 Residents in NL (2013)f 35.6 65 3.1 Residents in CH (2017)g 36.8 90.4 3.4 Residents in England (2018)h 29 62 2.7 population Daily distance [km] Daily travel time [min] No of trips per daya As far as the characteristics of the stay are concerned, one can notice that guests in the Ötztal valley choose car more often than guests in the other two regions. This holds in both seasons. The Zillertal, on the other hand, has the highest percentage of transit users, which can be attributed to a more extensive transit network, including a 32-km-long nar- row-gauge railway that runs through the valley. Traveling to the destination by private car leads to a high share of car trips on site. In contrast, train and coach travelers tend to use local public transport relatively frequently. Also, better knowledge of the destination has a favorable effect on choosing transit at the cost of car. f Figure 10 illustrates the temporal distribution of trips during one day (averaged) in sum- mer and winter seasons. In winter, two distinct peaks are apparent for all three modes, which clearly reflects how the mobility patterns are associated with the opening hours of ski resorts. Driving in summer exhibits similar morning and evening peaks as it is in win- ter, whereas walking trips are more uniformly distributed over the course of a day. In sum- mer, unlike in winter, afternoon return trips on transit are spread over several hours and do not form a peak as they do in the morning. Figure 11 presents the modal split with respect to the trip purpose. A distinct pat- tern can be observed, in particular in summer, in mode choice preferences between for traveling to social activities (e.g. Transport mode choice of the respondents The descriptive analysis of tourists’ choices in this paper is limited to transport mode choice. In summer, the share of trips made by car in the sample hits the highest value of almost 50%, followed by walking with 40%, transit (9%) and cycling (1%). In winter, walk- ing is the dominating mode reaching 47%, followed by driving and transit, with 36% and 17% respectively. Cycling was not reported in winter. Figures 8 and 9 provide informative insights into the relationship between transport mode choice and sociodemographic and stay-related variables. For instance, tourists over 65 years old choose transit more often than other age groups, especially in winter. Those who frequently use cars at home are also more likely to choose cars on vacation (in sum- mer). Interestingly, good health and lack of physical disabilities results in more walking trips in winter, but less in summer. 1 3 1 1 3 1499 Transportation (2022) 49:1465–1516 Table 7 Mobility rates of the surveyed sample of tourists and the corresponding rates in countries where the majority of guests in Tyrol originates from. Values per day per person (mobile and non-mobile persons together) Table 7 Mobility rates of the surveyed sample of tourists and the corresponding rates in countries where the majority of guests in Tyrol originates from. Values per day per person (mobile and non-mobile persons together) together) a Excluding cable car trips b Values for summer and winter respectively c Bundesministerium für Verkehr, Innovation und Technologie (2016) d Köll and Bader (2011) e Bundesministerium für Verkehr und digitale Infrastruktur (2018) f Hoogendoorn-Lanser et al. 4 It might however result from a small number of destinations for pursuing the planned activities, forcing families to travel far (e.g. only one ski resort nearby). Transport mode choice of the respondents going out, restaurant visit) and traveling to outdoor 1 3 Transportation (2022) 49:1465–1516 1500 and sport activities. In the first case, walking is the dominant mode, whereas in the latter, and generally with the increasing need to transport any kind of luggage or spe- cific items (trekking poles, climbing or water sports equipment, etc.), the share of car trips escalates. An exception to this rule is skiing, where a relatively high proportion of transit trips can be explained by the high-quality ski-bus services offered in winter, tailored specifically to skiers’ and snowboarders’ needs. i Figures 12 and 13 illustrate how long the trips undertaken by tourists are by each mode (data at the level of individual observations, not respondents). Tourists in sum- mer travel longer distances and spend more time traveling than in winter, regardless of which transport mode they choose (see also Table 7). Joint travel Joint travel accounts for a very high share of all trips in the sample. Out of 3120 trips, 3048 trips (98%) were made with an accompanying person (not necessarily a relative or household member), 2671 trips (86%) with at least one household member (this includes e.g. grandparents living with the family), whereas 2594 trips (83%) were made with immediate family members, i.e. a spouse or children. Figure 14 illustrates the relationship between the chosen mode and the family com- position on a trip, as well as between the chosen mode and the number of accompanying household members. In principle, as the group size increases, the preference for walking decreases and instead the preference for driving increases. Figure 15 illustrates the relationship between trip distance and family composition. As long as traveling with children clearly influences the choice of transport mode, it does not seem to influence the choice of destination much. Parents with and without children, alone and with spouses, undertake longer trips equally often. Apparently, parents do not avoid traveling with kids to distant locations within the vacation region. They adapt the mode choice in the first place, but the final destination choice remains unaffected.4 Impact of weather 10 Number of trips made by a given mode depending on time of day Fig. 11 Chosen mode depending on current trip purpose (i.e. the activity following the current trip) Impact of weather Respondents were asked to assess the impact of weather on their activity and transport mode choices. In 5.92% cases in summer and 1.52% cases in winter, they were forced to choose an alternative activity, whereas only in 0.98% cases in summer and 1.8% cases in winter, they had to resort to another means of transport due to unfavorable weather con- ditions. These statements show a very low impact of weather on tourists’ choices and suggest that tourists are determined to follow the vacation schedule (that they probably diligently prepared beforehand) no matter the weather. If we combine the responses based on personal weather perception with real meas- urement data from weather stations located near the starting points of the trips, a similar picture emerges (Table 8). Only in up to 5% observations, when it rained in summer, respondents declared to have chosen another transport mode than planned.i This is confirmed in Fig. 16 illustrating the transport mode choice as a function of precipitation. Visitors seem to be very indifferent to precipitation—in fact, the share of trips made on foot is higher on rainy days in summer and snowy days in winter than on 1 3 Transportation (2022) 49:1465–1516 1501 Fig. 8 Chosen mode depending on the sociodemographic characteristics Fig. 8 Chosen mode depending on the sociodemographic characteristics 3 3 Transportation (2022) 49:1465–1516 1502 Fig. 9 Chosen mode depending on characteristics of the stay and travel to the destination Fig. 9 Chosen mode depending on characteristics of the stay and travel to the destination Transportation (2022) 49:1465–1516 1503 p ( ) days with no precipitation. This positive effect on walking raises doubts about the plau- sibility of this outcome (however a similar finding was reported also by Saneinejad et al. (2012)). It is postulated that studying the activity, destination and mode choice jointly may explain the underlying dependencies more precisely. Not accounting for interfer- ence of weather on the activity and destination choice, can lead to questionable results Fig. 10 Number of trips made by a given mode depending on time of day Fig. 11 Chosen mode depending on current trip purpose (i.e. the activity following the current trip) Fig. 10 Number of trips made by a given mode depending on time of day Fig. 10 Number of trips made by a given mode depending on time of day Fig. Fig. 11 Chosen mode depending on current trip purpose (i.e. the activity following the current trip) days with no precipitation. This positive effect on walking raises doubts about the plau- sibility of this outcome (however a similar finding was reported also by Saneinejad et al. (2012)). It is postulated that studying the activity, destination and mode choice jointly may explain the underlying dependencies more precisely. Not accounting for interfer- ence of weather on the activity and destination choice, can lead to questionable results at the mode choice level—an issue also raised by Liu et al. (2015). 3 1 Transportation (2022) 49:1465–1516 1504 Fig. 12 Length of trips [km] depending on chosen mode Fig. 13 Duration of trips [min] depending on chosen mode Fig. 12 Length of trips [km] depending on chosen mode Fig. 12 Length of trips [km] depending on chosen mode Fig. 12 Length of trips [km] depending on chosen mode Fig. 13 Duration of trips [min] depending on chosen mode Fig. 13 Duration of trips [min] depending on chosen mode Fig. 13 Duration of trips [min] depending on chosen mode Conclusion The paper reports on the motivation for, design and administration of a data collection effort on tourists’ travel behavior at vacation destinations, as well as on basic descriptive results from the sample collected. Given the rising leisure mobility of modern societies (Larsen et al. 2006), there is a growing demand for understanding people’s travel behav- ior in discretionary travel and mobility at non-home locations. This study responds to that demand and contributes to the development of quantitative research on the transport behav- ior of tourists at the same level as that of residents. The presented dataset is unique in that it describes tourists’ actual observed behavior (revealed preferences) at a disaggregate level rather than behavioral intentions (stated preferences), making it a solid basis for developing 1 3 1 Transportation (2022) 49:1465–1516 1505 behavioral choice models, as is typically done for local residents for which census and national travel survey data are available. This paper provides researchers, transport plan- ners and policy-makers with a groundwork for further analytical and modeling work on tourist travel behavior and should ultimately foster better design of transport policies in tourist regions. The data collection process builds to some extent on the design of established surveys of everyday travel behavior. However, it is applied in a novel context and to an unconven- tional population of tourist visitors staying for a short time at a destination for vacation Fig. 14 Chosen mode depending on family composition during the trip Fig. 15 Distance traveled depending on family composition Fig. 14 Chosen mode depending on family composition during the trip Fig. 14 Chosen mode depending on family composition during the trip Fig. 14 Chosen mode depending on family composition during the trip Fig. 15 Distance traveled depending on family composition Fig. 15 Distance traveled depending on family composition Fig. 15 Distance traveled depending on family composition behavioral choice models, as is typically done for local residents for which census and national travel survey data are available. This paper provides researchers, transport plan- ners and policy-makers with a groundwork for further analytical and modeling work on tourist travel behavior and should ultimately foster better design of transport policies in tourist regions. The data collection process builds to some extent on the design of established surveys of everyday travel behavior. Conclusion However, it is applied in a novel context and to an unconven- tional population of tourist visitors staying for a short time at a destination for vacation purposes. We postulate that surveys of tourist travel behavior should collect disaggregate data on trips and activities of single respondents using a diary-based approach, if possible accompanied by GPS tracking, and should account for additional variables, beyond travel time, travel cost and level of service, which are typical predictors of home- and work- related travel choices (Frank et al. 2007). The survey instrument must allow the analyst to precisely differentiate between activities involving movement (e.g. cycling for pleasure) 1 3 3 3 Transportation (2022) 49:1465–1516 1506 and movements to activities (e.g. cycling to a supermarket), i.e. displacement to a place where the activity is performed, which can be confusing in a vacation context, where many trips tend to have a strong recreational or sporting character and may serve not only reloca- tion purposes.fif Table 8 Impact of (perceived) weather on mode choice Variable Value Summer Winter Precip. > 0 Precip. = 0 Precip. > 0 Precip. = 0 Impact of weather on mode choice 1st choice transport mode (as planned) 95.1% 99.6% 100.0% 98.0% 2nd choice transport mode (plan B) 4.9% 0.4% 0.0% 2.0% Fig. 16 Mode choice of tourists depending on precipitation Table 8 Impact of (perceived) weather on mode choice Variable Value Summer Winter Precip. > 0 Precip. = 0 Precip. > 0 Precip. = 0 Impact of weather on mode choice 1st choice transport mode (as planned) 95.1% 99.6% 100.0% 98.0% 2nd choice transport mode (plan B) 4.9% 0.4% 0.0% 2.0% Table 8 Impact of (perceived) weather on mode choice Fig. 16 Mode choice of tourists depending on precipitation Fig. 16 Mode choice of tourists depending on precipitation and movements to activities (e.g. cycling to a supermarket), i.e. displacement to a place where the activity is performed, which can be confusing in a vacation context, where many trips tend to have a strong recreational or sporting character and may serve not only reloca- tion purposes.fif and movements to activities (e.g. cycling to a supermarket), i.e. 1 3 Conclusion f • Whether they purchased any kind of regional guest card in the summer season entitling them to free or discounted use of public transportation.i f • Whether they purchased any kind of regional guest card in the summer season entitling them to free or discounted use of public transportation.i • The exact age of children (or at least stratified 0–6, 7–15 and 16–20 years) so that it would correspond to single fare categories and allow for more accurate cost calcula- tions. Sample size is another clear limitation of the study. With 849 respondents, the sample is within the range of comparable studies on tourist mobility. However, it is rather small (approx. 2.5%) in relation to the average of 34,000 tourists staying in all three regions at any time (given over 85,000 beds available and an average load factor of around 40%). It is also not representative, as most of the data was collected using convenience sampling and the response rate to questionnaires distributed in the hotels was very low (which would otherwise have made it possible to control the sampling process). It must be borne in mind that results based on a sample of this kind cannot be easily generalized to larger or different study populations. As discussed in Sect. Descriptive analysis, a large share of trips within the tourist desti- nation are shorter than 2 km and are made on foot. The accuracy of distance and travel time calculation for short trips using Google Maps API might become questionable, since fac- tors other than pure walking speed5 come into play. Walking speed may, besides personal factors (age, gender, fitness, trip purpose, mood, etc.), depend also on physical conditions such as carried items (sports equipment, baby carriage, groceries), altitude difference, side- walk surface, or weather conditions (snow layer). This might have implications for pro- spective choice modeling work, given the considerable share of short walking trips in the current dataset. Future studies should consider collecting even more precise data to account for this. 5 Google does not disclose what speed they use in their routing system. As long as transit is concerned, it is based on timetable; car speeds are based on live traffic data, but the assumptions behind walking and cycling are unknown. However, one can observe that the speed varies depending on the elevation difference. Conclusion displacement to a place where the activity is performed, which can be confusing in a vacation context, where many trips tend to have a strong recreational or sporting character and may serve not only reloca- tion purposes.fif We experienced difficulties in establishing effective cooperation with the accommoda- tion providers, both without and with the support of the local DMOs. This affected the choice of the survey methods and, consequently, led to a loss of control over the sampling process (change from distributed PAPI questionnaires to face-to-face CAPI interviews). The study also revealed an ambiguous effect of incentives. Prospective studies should secure a larger budget and test an effect of higher monetary incentives. In particular, a suc- cessful implementation of a survey conducted in close cooperation with accommodation providers and accompanied with substantial incentives for the participants and the accom- modations would be of value to the scientific community.f i The paper also analyzes and compares seasonal differences between mode choice behav- ior of tourists in summer and winter. The basic descriptive results suggest that the effects of group composition, trip purpose, weather, information about the destination might be of non-negligible magnitude. Thus, although accounting for additional variables leads increases data collection costs, complexity, and response burden, it is strongly advised that this be done as early as at the survey design stage. The results of a modeling work based on this study revealed several limitations of the dataset. Some specific information that could have contributed to a deeper understanding of tourists’ mobility choices was not collected in the survey. These include, in particular, the following: 1 3 1 3 Transportation (2022) 49:1465–1516 1507 • Whether respondents had to carry luggage (e.g. climbing equipment, snow sleds for kids, stroller) on their trip to the activity start location and whether it was bulky and troublesome. • Whether respondents had to carry luggage (e.g. climbing equipment, snow sleds for kids, stroller) on their trip to the activity start location and whether it was bulky and troublesome. • Whether they brought their own bicycles or e-bikes with them.f • Whether their hotel offered them bicycles or e-bikes for rent or free of charge. Appendix See Figs. 17, 18, 19, 20. See Figs. 17, 18, 19, 20. 5 Google does not disclose what speed they use in their routing system. As long as transit is concerned, it is based on timetable; car speeds are based on live traffic data, but the assumptions behind walking and cycling are unknown. However, one can observe that the speed varies depending on the elevation difference. 3 Transportation (2022) 49:1465–1516 1508 Fig. 17 PAPI questionnaire—front and back cover Fig. 17 PAPI questionnaire—front and back cover Fig. 18 PAPI questionnaire—page 1 Fig. 18 PAPI questionnaire—page 1 Fig. 18 PAPI questionnaire—page 1 1 3 1 3 Transportation (2022) 49:1465–1516 1509 Fig. 19 PAPI questionnaire—page 2 Fig. 19 PAPI questionnaire—page 2 Fig. 19 PAPI questionnaire—page 2 1 3 Fig. 20 PAPI questionnaire—page 3 Fig. 20 PAPI questionnaire—page 3 Fig. 20 PAPI questionnaire—page 3 3 3 1510 Transportation (2022) 49:1465–1516 Acknowledgements The first author would like to thank the Faculty of Engineering Sciences at the Uni- versity of Innsbruck for supporting him in the data collection process with need-based scholarships (No. 1092191, No. 1354749). We would also like to thank Emma Komarek, former student assistant in the Unit of Intelligent Transport Systems at the University of Innsbruck, for her work on data digitalization and data cleansing. Author’s contributions BB: Conceptualization, Methodology, Formal analysis, Investigation, Writing— Original Draft, Visualization. MM: Conceptualization, Resources, Writing—Review & Editing, Supervi- sion. KWA: Conceptualization, Writing—Review & Editing, Supervision. Author’s contributions BB: Conceptualization, Methodology, Formal analysis, Investigation, Writing— Original Draft, Visualization. MM: Conceptualization, Resources, Writing—Review & Editing, Supervi- sion. KWA: Conceptualization, Writing—Review & Editing, Supervision. Funding Open access funding provided by University of Innsbruck and Medical University of Innsbruck Availability of data and material The survey data along with a codebook and metadata are currently being prepared for publication in a repository. Availability of data and material The survey data along with a codebook and metadata are currently being prepared for publication in a repository. Declarations Conflict of interest On behalf of all authors, the corresponding author declares no conflict of interest. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Com- mons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. 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https://openalex.org/W2064803625	https://dash.harvard.edu/bitstream/1/41511302/1/78341%202007_physrevlet_utada.pdf	English	null	Dripping to Jetting Transitions in Coflowing Liquid Streams	Physical review letters	2,007	cc-by	4,139	Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA Permanent link http://nrs.harvard.edu/urn-3:HUL.InstRepos:41511302 Citation Utada, Andrew S., Alberto Fernandez-Nieves, Howard A. Stone, and David A. Weitz. 2007. “Dripping to Jetting Transitions in Coflowing Liquid Streams.” Physical Review Letters99 (9): 094502. https://doi.org/10.1103/PhysRevLett.99.094502. DOI: 10.1103/PhysRevLett.99.094502 A liquid forced through an oriﬁce will ultimately break into drops through one of two mechanisms. At slow ﬂows, the emerging liquid drips from the oriﬁce, whereas at faster ﬂows, the liquid forms a thin stream that breaks into drops away from the oriﬁce; these are the dripping and jetting regimes. This behavior is familiar to anyone who has slowly increased the ﬂow rate of water at a kitchen faucet. In this case, dripping occurs at low ﬂow rates where surface tension causes the water to form drops at the tap; the hanging drops detach when the gravitational force exceeds surface tension forces. At higher ﬂow rates jetting occurs when the inertial forces of the water exceed surface tension forces [1,2]. The dripping-to-jetting transition is sharp if the liquid viscosity is large compared to water [2], whereas the transition initially becomes chaotic [1,3–5] before jet- ting for lower viscosity ﬂuids. The jets eventually break into drops due to the Rayleigh-Plateau instability [6,7]. W in; these parameters describe, respectively, the magni- tude of the viscous shear forces from the outer liquid and the inertial forces from the inner liquid compared to sur- face tension forces. We observe two distinct jetting re- gimes with signiﬁcant differences in jet shape and the mechanism controlling drop size. g p Our experimental device is made of two coaxially aligned capillary tubes. The inner capillary tube is cylin- drical, with a tip tapered to an inner diameter of dtip ’ 20 !m and an outer diameter of 30 !m. The outer capil- lary tube is square; coaxial alignment of the tubes is achieved by matching the outer diameter of the untapered portion of the inner capillary to the inner dimension of the square capillary, D ! 1 mm, as shown in Fig. 1(a). At this length scale, which is below the capillary length, the effects of gravity are negligible. Although the ﬂow in the square tube is not axisymmetric, since the tip is centered and dtip=D " 0:02, the local ﬂow around the tip should be approximately axisymmetric. For experiments requiring a larger velocity of the outer ﬂuid, we place a second cylin- drical capillary with inner diameter of 200 !m inside the outer square capillary, surrounding the tip; since this extra capillary has a smaller cross section than the square one, the outer ﬂuid achieves a higher average velocity. Share Your Story The Harvard community has made this article openly available. Please share how this access benefits you. Submit a story . The Harvard community has made this article openly available. Please share how this access benefits you. Submit a story . Accessibility P H Y S I C A L R E V I E W L E T T E R S week ending 31 AUGUST 2007 week ending 31 AUGUST 2007 PRL 99, 094502 (2007) Dripping to Jetting Transitions in Coﬂowing Liquid Streams Andrew S. Utada,1 Alberto Fernandez-Nieves,1,2 Howard A. Stone,1 and David A. Weitz1,3,* 1Harvard School of Engineering and Applied Sciences, Cambridge, Massachusetts 02138, USA 2INEST Group, Philip Morris USA, Richmond, Virginia 23224, USA 3The Physics Department, Harvard University, Cambridge, Massachusetts 02138, USA (Received 21 October 2006; published 28 August 2007) A liquid forced through an oriﬁce into an immiscible ﬂuid ultimately breaks into drops due to surface tension. Drop formation can occur right at the oriﬁce in a dripping process. Alternatively, the inner ﬂuid can form a jet, which breaks into drops further downstream. The transition from dripping to jetting is not understood for coﬂowing ﬂuid streams, unlike the case of drop formation in air. We show that in a coﬂowing stream this transition can be characterized by a state diagram that depends on the capillary number of the outer ﬂuid and the Weber number of the inner ﬂuid. PACS numbers: 47.61.Jd, 47.20.Dr, 47.27.nf, 47.55.db DOI: 10.1103/PhysRevLett.99.094502 DOI: 10.1103/PhysRevLett.99.094502 DOI: 10.1103/PhysRevLett.99.094502 Although the surrounding walls are closer in this case, the jet diame- ter is # dtip, and dtip=D " 0:1; thus, we expect wall effects to be small. We use deionized water and different viscosity polydimethylsiloxane (PDMS) oils; interchanging the oil and water enables us to vary the viscosity ratio, "in="out, from 0.01 to 10, where "in and "out are the viscosities of the inner and outer ﬂuids, respectively. The surface tension between PDMS oil and water can be lowered from 40 to 4 mN=m by adding 60 mM sodium dodecyl sulfate (SDS) to the water. This high concentration of SDS reduces surface tension gradients across the jets. While both dripping and jetting must occur when a liquid is injected in a second immiscible liquid [8–10], the mechanism of droplet formation changes due to the presence of the surrounding viscous liquid [11–14]. Drop formation has rich dynamics [15,16] that are affected by many parameters such as the average velocities of both liquids, their viscosities and densities, surface tension, and the surface chemistry and device geometry [17]. Two- phase drop formation is important to applications in micro- ﬂuidics [18,19] such as ﬂow focusing [15,20] and in liquid- gas systems [21], selective withdrawal for coating particles [22], and extrusion emulsiﬁcation [23]. Because of its importance, drop formation in two ﬂuids has been widely studied [8–10,24,25]; however, complete control over the two-phase ﬂow behavior requires a detailed understanding of the dripping-to-jetting transition, and a uniﬁed view of this transition is still lacking [26]. In this Letter, we use a microcapillary device to study the transition between dripping and jetting in a two-phase coﬂowing stream. The behavior is characterized by a state diagram that depends on both the capillary number of the outer ﬂuid, Cout, and the Weber number of the inner ﬂuid, In coﬂowing ﬂuids, dripping occurs at low ﬂow rates of both ﬂuids and is characterized by the periodic formation of individual drops that pinch-off from the tip [Fig. 1(b)]. We observe two distinct classes of transitions from drip- 094502-1 2007 The American Physical Society 0031-9007=07=99(9)=094502(4) P H Y S I C A L R E V I E W L E T T E R S week ending 31 AUGUST 2007 FIG. 1. (a) Device geometry showing the tapered inner capil- lary in the outer square capillary. (b) Image of the dripping regime. DOI: 10.1103/PhysRevLett.99.094502 (c) Image of a narrowing jet generated by increasing qout above a threshold while keeping qin constant, with "in="out ! 0:1. (d) Image of a widening jet generated by increasing qin above a threshold while keeping qout constant, with "in="out ! 0:1. Images (b)–(d) were taken with a high-speed camera (walls not shown). Scale bar applies to (b)–(d). PRL 99, 094502 (2007) [27]. The diameter of the detaching drop decreases as qout increases; ultimately, a critical shear stress is reached where the emerging liquid is stretched into a narrowing jet [Fig. 1(c)]. The diameter of this jet is initially equal to dtip and decreases as the liquid moves downstream until the stress gradient across the jet relaxes. This is similar to ﬂow focusing in microﬂuidic devices [15,20] and in model calculations [28]. We measure the ﬁnal downstream diameter of the jets and resulting drops from high-speed movies. We compare these data to calculations of the steady-state diameter of the jet, djet, obtained by solving for the motion of two coaxially ﬂowing liquids with constant shape under Stokes ﬂow: "r2u ! rp, where p is the pressure. By relating the mean velocities of both ﬂuids to the ﬂow rates, we obtain a function [29] where the dominant term is qin=qout " 2%djet=D&2. Solving for djet=D, we obtain the scaled jet diameter as a function of the ﬂow rate ratio. This relation (solid line) agrees well with the measured jet diameters, as shown in Fig. 2. FIG. 1. (a) Device geometry showing the tapered inner capil- lary in the outer square capillary. (b) Image of the dripping regime. (c) Image of a narrowing jet generated by increasing qout above a threshold while keeping qin constant, with "in="out ! 0:1. (d) Image of a widening jet generated by increasing qin above a threshold while keeping qout constant, with "in="out ! 0:1. Images (b)–(d) were taken with a high-speed camera (walls not shown). Scale bar applies to (b)–(d). We can also calculate the drop diameter by assuming that it is related to djet. For static ﬂuid cylinders suspended in another immiscible liquid, the wavelength of the fastest growing mode of the Rayleigh-Plateau instability is pro- portional to the diameter of the cylinder itself [30]. DOI: 10.1103/PhysRevLett.99.094502 In turn, the volume contained in one wavelength of this mode, $, on the cylinder, %d2 jet$=4, is approximately equal to the volume of the drop that is formed when the cylinder breaks. The most unstable mode is a function of viscosity ratio; in this experiment, "in="out ! 0:1 and $ ! 5:48djet [30]. We solve for ddrop as a function of djet, assuming that this argument applies for nearly uniformly translating jets, giving ddrop " 2djet. The calculated results (dotted line) agree very well with the measured drop sizes, as shown in Fig. 2. ping to jetting. The ﬁrst is driven by the ﬂow rate of the outer ﬂuid; as it is increased, drops formed at the tip decrease in size until a jet is formed, whereupon drop breakup occurs downstream at the end of the thin jet [Fig. 1(c)]. The second class of transition is driven by the ﬂow rate of the inner ﬂuid; as it is increased, the dripping drop is pushed downstream and is ultimately pinched off at the end of the resultant jet [Fig. 1(d)]. The ﬁrst class of dripping-to-jetting transition is charac- terized by a jet that thins as it moves downstream. In the dripping regime, the diameter of the drop ﬁrst decreases as the ﬂow rate of the outer liquid, qout, increases. When the drop diameter becomes approximately equal to dtip, there is a spontaneous transition to jetting. The diameter of the jet decreases with distance downstream but ultimately reaches a constant value as shown in Fig. 1(c). Still farther downstream, the jet develops undulations driven by the Rayleigh-Plateau instability; these grow larger and ulti- mately drive the formation of drops whose diameter is only slightly larger than that of the cylindrical jet itself. In this regime, it is the viscous drag from the ﬂow of the outer ﬂuid that drives drop formation. In the dripping regime, the growing droplet experiences two competing forces: viscous drag pulling it downstream and forces due to surface tension holding it to the tip. Initially, surface tension dominates but the drag forces eventually become comparable as the droplet grows. This force balance is given by: "outuoutddrop $ #dtip, where uout is the mean velocity of the outer liquid and # is the surface tension FIG. 2 (color online). DOI: 10.1103/PhysRevLett.99.094502 For widening jets, the inner liquid is injected at a much larger velocity than the average velocity of the outer liquid. The large shear at the interface due to the velocity difference decelerates the jet, causing it to widen. mation encompasses three steps: advection and growth of the droplet at the end of the jet, drop pinchoff, and ﬁnally retraction of the neck. The advection of the growing drop- let by the viscous drag is the slowest of these steps; images illustrating the distance the jet is advected are shown in the lower inset of Fig. 3. Since the velocity of the inner liquid is much larger than the advection velocity, we expect ' to scale as the advected distance divided by the advection velocity. This distance is typically $ddrop while the veloc- ity is $uout, which leads to ddrop $ %qin=uout&1=2. This agrees with the exponent of 0.46 determined from a ﬁt of the data for "in="out ! 0:1, as shown by the solid line in Fig. 3. While we limit our discussion of drop scaling to "in="out ! 0:1, this result is notable because drop size is set by viscous drag forces on the growing droplet, whereas in the limit of "in="out ' 1, jet and drop diameters are set solely by the tip diameter, regardless of the jet velocity. y y p g j y To unify these two distinct jetting transitions, we con- sider the balance of forces on the droplet immediately before the dripping-to-jetting transition. In the ﬁrst regime, where the jets become thinner, the viscous shear stresses on the drop must be balanced by surface tension; this balance is given by the capillary number of the outer ﬂuid, Cout ! "outuout=#, at the transition. When Cout is small, surface tension dominates, forcing the system to drip. By contrast, when Cout ( O%1&, the shear stress on the droplet is large enough to overcome surface tension, leading to jetting. In the second regime, where the jets become wider, the iner- tial forces of the inner ﬂuid must be balanced by the surface tension forces; this balance is given by the inner Weber number, W in ! &indtipu2 in=#. When W in is small, surface tension dominates, forcing the system to drip. By contrast, The large qualitative shape differences between the widening and the thinning jets indicate a different balance of dominant forces. DOI: 10.1103/PhysRevLett.99.094502 Experimentally measured djet (!) and ddrop (") scaled by D as a function of qin=qout for the narrowing jets with "in="out ! 0:1. The solid line is the prediction from the model for djet with no ﬁtting parameters. The dashed line is the predicted result assuming ddrop " 2djet from the Rayleigh- Plateau instability as described in the text. The inner and outer Reynolds numbers, Rin ! &indtipuin "in and Rout ! &outDuout "out , vary from $7–70 and $15–50, respectively. FIG. 2 (color online). Experimentally measured djet (!) and ddrop (") scaled by D as a function of qin=qout for the narrowing jets with "in="out ! 0:1. The solid line is the prediction from the model for djet with no ﬁtting parameters. The dashed line is the predicted result assuming ddrop " 2djet from the Rayleigh- Plateau instability as described in the text. The inner and outer Reynolds numbers, Rin ! &indtipuin "in and Rout ! &outDuout "out , vary from $7–70 and $15–50, respectively. 094502-2 094502-2 P H Y S I C A L R E V I E W L E T T E R S week ending 31 AUGUST 2007 PRL 99, 094502 (2007) FIG. 3. Measured drop diameter ()) as a function of qin=uout for widening jets with "in="out ! 0:1. The slope of the best ﬁt, shown as the solid line, is 0.46. Upper inset: Drop diameter as a function of uout, for constant qin () and as a function of qin, for constant qout ("). Lower inset: Image (top) just after drop pinch- off, where the jet has retracted due to surface tension and just before the next drop pinches-off (bottom). The end of the jet travels a distance indicated by the arrow in this time interval. Here, Rin and Rout vary from $60–200 and $1–60, respec- tively. The second dripping-to-jetting transition is character- ized by a dramatically different shape; the jet diameter increases along its length instead of decreasing [Fig. 1(d)]. In this case, the transition from dripping to jetting is no longer sudden; instead, there is an intermediate dripping regime as qin increases where the growing droplets move downstream while still connected to the ﬂuid in the tip through a ﬂuid neck. However, as soon as the drop pinches off, the neck retracts completely back to the tip, whereupon the process repeats itself. DOI: 10.1103/PhysRevLett.99.094502 As qin is increased still further, this intermediate regime transitions fully into jetting, where we deﬁne jetting as the incomplete retraction of the ﬂuid neck. The jet ultimately develops undulations that lead to drop formation. FIG. 3. Measured drop diameter ()) as a function of qin=uout for widening jets with "in="out ! 0:1. The slope of the best ﬁt, shown as the solid line, is 0.46. Upper inset: Drop diameter as a function of uout, for constant qin () and as a function of qin, for constant qout ("). Lower inset: Image (top) just after drop pinch- off, where the jet has retracted due to surface tension and just before the next drop pinches-off (bottom). The end of the jet travels a distance indicated by the arrow in this time interval. Here, Rin and Rout vary from $60–200 and $1–60, respec- tively. This second class of dripping-to-jetting transition is controlled by different forces and thus exhibits different behavior. The control parameter qin does not affect drop size in the dripping regime; instead drop size depends on the viscous drag, which depends on qout. Since qout is held constant, qin affects how quickly the droplets reach the critical size where the viscous drag and surface tension forces balance each other. However, qin is proportional to the momentum of the inner ﬂuid, which acts cooperatively with the viscous drag from the outer ﬂuid to drive the transition to the intermediate regime, where the droplet moves downstream, though still connected to the ﬂuid in the tip through a ﬂuid neck. After the drop pinches off, surface tension pulls the neck completely back into the tip. At even larger qin, the inertia of the injected inner liquid becomes sufﬁciently large that the position of drop growth and pinch off is pushed away from the tip; the large inertial forces within the jet prevent the ﬂuid neck from retracting completely back to the tip after drop pinch off. There is, however, a slight retraction at the end of the jet due to surface tension, which causes the end to become spherical and move slightly upstream. Viscous drag from the outer ﬂuid then pulls the growing droplet downstream beginning the process again. The signiﬁcant shape differences be- tween these jets and the thinning jets [see Figs. 1(c) and 1(d)] are due to the large velocity difference between the inner and outer liquids. DOI: 10.1103/PhysRevLett.99.094502 4 mN=m. Typical error bars for different surface tensions are shown. [3] R. Shaw, The Dripping Faucet as a Model Chaotic System (Aerial, Santa Cruz, CA, 1984). [4] B. Ambravaneswaran, S. D. Phillips, and O. A. Basaran, Phys. Rev. Lett. 85, 5332 (2000). [5] P. Coullet, L. Mahadevan, and C. S. Riera, J. Fluid Mech. 526, 1 (2005). [6] J. Plateau, Acad. Sci. Bruxelles Me´m. 23, 5 (1849). [7] L. Rayleigh, Proc. R. Soc. London 29, 71 (1879). [8] G. F. Scheele and B. J. Meister, AIChE J. 14, 9 [9] B. J. Meister and G. F. Scheele, AIChE J. 15, 689 (1969); 15, 700 (1969). [10] J. R. Richards, A. N. Beris, and A. M. Lenhoff, Phys. Fluids 7, 2617 (1995). when W in ( O%1&, the inertial forces are large enough to overcome surface tension, leading to jetting. To generalize this result, we vary the viscosity ratio between 0.01 and 10 and lower surface tension from 40 to 4 mN=m for "in="out ! 10. We plot a state diagram showing dripping (closed symbols) and jetting (open symbols) as a function of Cout and W in. Dripping occurs when both Cout and W in are small; however, there is a distinct transition when either increases, as shown by the wide gray line in Fig. 4. The good collapse of the data reveals that in the coﬂowing stream the sum of the forces exerted on the droplet must ultimately overcome surface tension forces to cause the transition; this can be summarized by Cout + W in " O%1&. [11] D. F. Zhang and H. A. Stone, Phys. Fluids 9, 2234 (1997). [12] J. R. Lister and H. A. Stone, Phys. Fluids 10, 2758 (1998). [13] W. W. Zhang and J. R. Lister, Phys. Rev. Lett. 83, 1151 (1999). [14] I. Cohen et al., Phys. Rev. Lett. 83, 1147 (1999). [15] S. L. Anna, N. Bontoux, and H. A. Stone, Appl. Phys. Lett. 82, 364 (2003). [16] P. Garstecki, M. J. Fuerstman, and G. M. Whitesides, Phys. Rev. Lett. 94, 234502 (2005). [17] J. Eggers, Rev. Mod. Phys. 69, 865 (1997). [18] M. Joanicot and A. Ajdari, Science 309, 887 (2005). [19] H. A. Stone, A. D. Stroock, and A. Ajdari, Annu. Rev. Fluid Mech. 36, 381 (2004). [20] A. M. Gan˜a´n-Calvo and J. M. Gordillo, Phys. Rev. Lett. 87, 274501 (2001). DOI: 10.1103/PhysRevLett.99.094502 These differences even change the mechanism controlling drop size, which is highlighted by the fact that in the widening regime ddrop ! 2djet. Representative data sets illustrate trends in drop size as a function of qin and uout, as shown in the upper inset of Fig. 3. To elucidate the drop-size scaling from these scat- tered data, we use mass conservation to relate drop diame- ter to the control parameters: the volume of a drop is V ! qin', where ' is the drop formation time. Drop for- 094502-3 P H Y S I C A L R E V I E W L E T T E R S week ending 31 AUGUST 2007 week ending 31 AUGUST 2007 PRL 99, 094502 (2007) FIG. 4. State diagram of the dripping-to-jetting transition in a coﬂowing stream as a function of Cout and W in. Filled symbols represent dripping while open symbols represent jetting. Each shape is a different viscosity ratio, surface tension, or geometry. Surface tension is 40 mN=m unless otherwise stated. Square: "in="out ! 0:01. Diamond: "in="out ! 0:01, with the extra capillary tube to increase uout. Hexagon: "in="out ! 0:01. Circle: "in="out ! 0:01, with the extra capillary tube to increase uout. Pentagon: "in="out ! 1. Triangle: "in="out ! 10. Star: "in="out ! 10 and # ! 4 mN=m. Typical error bars for different surface tensions are shown. over emulsions generated with shear ﬂows. Moreover, although we consider only Newtonian ﬂuids, the basis for this transition may also clarify similar processes involving non-Newtonian ﬂuids. This work was supported by the NSF (No. DMR- 0602684) and the Harvard MRSEC (No. DMR-0213805). A. F.-N. thanks the University of Almeria. *weitz@seas.harvard.edu [1] C. Clanet and J. C. Lasheras, J. Fluid Mech. 383, 307 (1999). [2] B. Ambravaneswaran et al., Phys. Rev. Lett. 93, 034501 (2004). FIG. 4. State diagram of the dripping-to-jetting transition in a coﬂowing stream as a function of Cout and W in. Filled symbols represent dripping while open symbols represent jetting. Each shape is a different viscosity ratio, surface tension, or geometry. Surface tension is 40 mN=m unless otherwise stated. Square: "in="out ! 0:01. Diamond: "in="out ! 0:01, with the extra capillary tube to increase uout. Hexagon: "in="out ! 0:01. Circle: "in="out ! 0:01, with the extra capillary tube to increase uout. Pentagon: "in="out ! 1. Triangle: "in="out ! 10. Star: "in="out ! 10 and # ! DOI: 10.1103/PhysRevLett.99.094502 Our results identify the different jetting responses and highlight the importance of the coﬂowing viscous liquid in the dripping-to-jetting transition. For our experiments, the two jetting regimes result from a different balance of dominant forces. However, the two regimes can be inte- grated into a single picture that ultimately depends on the ability of these forces to deform the emerging liquid from a spherical drop to a cylindrical jet. In general, we expect the ﬂow response to depend on 5 dimensionless parameters, which can be chosen as (W in, Cout, Rin, Rout, uin=uout), where R is the Reynolds number. We ﬁnd that all of our data are captured by (W in, Cout) for moderate R. Thus our work provides a general framework for understanding a wide range of behavior, though clearly there are regions of the parameter space yet unexplored. A better understand- ing of this transition will provide a higher degree of control [21] B. K. H. Yen et al., Angew. Chem., Int. Ed. 44, 5447 (2005). [22] I. Cohen et al., Science 292, 265 (2001). [23] G. T. Vladisavljevic and R. A. Williams, Adv. Colloid Interface Sci. 113, 1 (2005). [24] B. J. Meister and G. F. Scheele, AIChE J. 13, 682 (1967). [25] J. R. Richards, A. N. Beris, and A. M. Lenhoff, Phys. Fluids A 5, 1703 (1993); J. R. Richards, A. M. Lenhoff, and A. N. Beris, Phys. Fluids 6, 2640 (1994). [26] C. Cramer, P. Fischer, and E. J. Windhab, Chem. Eng. Sci. 59, 3045 (2004). [27] P. B. Umbanhowar, V. Prasad, and D. A. Weitz, Langmuir 16, 347 (2000). [28] J. Meyer and D. Weihs, J. Fluid Mech. 179, 531 (1987). [29] A. S. Utada et al., Science 308, 537 (2005). [30] S. Tomotika, Proc. R. Soc. A 150, 322 (1935). 094502-4
https://openalex.org/W3162334094	https://www.researchsquare.com/article/rs-149050/latest.pdf	English	null	Circulating Tumor Cell Clusters Are Frequently Detected in Women with Early-Stage Breast Cancer	Cancers	2,021	cc-by	16,083	Dissemination of Circulating Tumo Occurs Early in Non฀metastatic Br Patients Carolina Reduzzi Fondazione IRCCS Istituto Nazionale dei Tumori https://orcid.org/ Serena Di Cosimo Fondazione IRCCS Istituto Nazionale dei Tumori Lorenzo Gerratana CRO IRCCS: Centro di Riferimento Oncologico Rosita Motta Fondazione IRCCS Istituto Nazionale dei Tumori Antonia Martinetti Fondazione IRCCS Istituto Nazionale dei Tumori Andrea Vingiani Fondazione IRCCS Istituto Nazionale dei Tumori Paolo D'Amico IEO: Istituto Europeo di Oncologia Youbin Zhang Northwestern University Feinberg School of Medicine Marta Vismara Fondazione IRCCS Istituto Nazionale dei Tumori Catherine Depretto Fondazione IRCCS Istituto Nazionale dei Tumori Gianfranco Scaperrotta Fondazione IRCCS Istituto Nazionale dei Tumori Secondo Folli Fondazione IRCCS Istituto Nazionale dei Tumori Giancarlo Pruneri Fondazione IRCCS Istituto Nazionale dei Tumori Massimo Cristofanilli Northwestern University Feinberg School of Medicine Maria Grazia Daidone Fondazione IRCCS Istituto Nazionale dei Tumori Vera Cappelletti (  vera.cappelletti@istitutotumori.mi.it ) Dissemination of Circulating Tumor Cell Clusters Occurs Early in Non฀metastatic Breast Cancer Patients Andrea Vingiani Fondazione IRCCS Istituto Nazionale dei Tumori Paolo D'Amico IEO: Istituto Europeo di Oncologia Catherine Depretto Fondazione IRCCS Istituto Nazionale dei Tumori Fondazione IRCCS Istituto Nazionale dei Tumori https://orcid.org/0000-0002-9709-0676 Research article Keywords: circulating tumor cell clusters, liquid biopsy, metastatic breast cancer, non-metastatic breast cancer, circulating tumor microemboli, size-based enrichment Posted Date: January 20th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-149050/v1 DOI: https://doi.org/10.21203/rs.3.rs-149050/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published on May 13th, 2021. See the published version at https://doi.org/10.3390/cancers13102356. Dissemination of circulating tumor cell clusters occurs early in non-metastatic breast cancer patients Carolina Reduzzi1,2,$, Serena Di Cosimo1,$, Lorenzo Gerratana2,3,4, Rosita M Carolina Reduzzi1,2,$, Serena Di Cosimo1,$, Lorenzo Gerratana2,3,4, Rosita Motta1, Antonia 1 Martinetti5, Andrea Vingiani6, Paolo D’Amico2,7, Youbin Zhang2, Marta Vismara1, Catherine 2 Depretto8, Gianfranco Scaperrotta8, Secondo Folli9, Giancarlo Pruneri G6, Massimo 3 Cristofanilli2, Maria Grazia Daidone1, Vera Cappelletti1,* 4 Martinetti5, Andrea Vingiani6, Paolo D’Amico2,7, Youbin Zhang2, Marta Vismara1, Catherine 2 Depretto8, Gianfranco Scaperrotta8, Secondo Folli9, Giancarlo Pruneri G6, Massimo 3 Cristofanilli2, Maria Grazia Daidone1, Vera Cappelletti1,* 4 1 Department of Applied Research and Technological Development, Fondazione IRCCS Istituto 5 Nazionale dei Tumori di Milano Milan Italy 6 1 Department of Applied Research and Technological Development, Fondazione IRCCS Istituto 5 Nazionale dei Tumori di Milano, Milan, Italy 6 2 Department of Medicine-Hematology and Oncology, Robert H. Lurie Comprehensive Cancer 7 Center, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA 8 tment of Medicine (DAME), University of Udine, Italy 3 Department of Medicine (DAME), University of Udine, Italy 9 4 Department of Medical Oncology, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, 10 Aviano, Italy 11 5 Medical Oncology Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, Italy 12 6 5 Medical Oncology Unit, Fondazione IRCCS Istituto Nazionale dei Tumori, 12 6 Pathology Department, Fondazione IRCCS Istituto Nazionale Tumori and University of Milan, 13 School of Medicine, Milan, Italy 14 7 New Drugs and Early Drug Development for Innovative Therapies Division, IEO, European 15 Institute of Oncology IRCCS, Milan, Italy; University of Milano, Department of Hematology and 16 Hemato-Oncology, Italy 17 8 Department of Radiology, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano, Milan, Italy 18 9 Breast Cancer Unit, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano, Milan, Italy 19 $Equally contributed 20 * Correspondence: 21 Vera Cappelletti 22 vera.cappelletti@istitutotumori.mi.it 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 6 Pathology Department, Fondazione IRCCS Istituto Nazionale Tumori and University of Milan, 13 School of Medicine, Milan, Italy 14 7 New Drugs and Early Drug Development for Innovative Therapies Division, IEO, European 15 Institute of Oncology IRCCS, Milan, Italy; University of Milano, Department of Hematology and 16 Hemato-Oncology, Italy 17 Abstract 39 Background: Metastatic spreading is promoted by cancer cell seeding from the primary tumor into 40 the bloodstream. In patients with metastatic breast cancer (MBC), the clinical relevance of circulating 41 tumor cell clusters (CTC-clusters) has been extensively reported, while their study in earlier stages is 42 limited. Several methods, besides the FDA-cleared CellSearch®, limited to the detection of 43 epithelial-enriched clusters, can be used for the detection of CTC-clusters. We hypothesize that 44 resorting to marker-independent approaches can improve CTC-cluster detection. 45 Background: Metastatic spreading is promoted by cancer cell seeding from the primary tumor into 40 the bloodstream. In patients with metastatic breast cancer (MBC), the clinical relevance of circulating 41 tumor cell clusters (CTC-clusters) has been extensively reported, while their study in earlier stages is 42 limited. Several methods, besides the FDA-cleared CellSearch®, limited to the detection of 43 epithelial-enriched clusters, can be used for the detection of CTC-clusters. We hypothesize that 44 resorting to marker-independent approaches can improve CTC-cluster detection. 45 Methods: Blood samples collected from healthy donors and spiked-in with tumor mammospheres, or 46 from BC patients, were processed for CTC-cluster detection with 3 technologies: CellSearch®, 47 CellSieve filters, ScreenCell® filters. The number of CTC-clusters was compared among the 48 technologies and analyzed in relation to patient characteristics and outcome. 49 Methods: Blood samples collected from healthy donors and spiked-in with tumor mammospheres, or 46 from BC patients, were processed for CTC-cluster detection with 3 technologies: CellSearch®, 47 CellSieve filters, ScreenCell® filters. The number of CTC-clusters was compared among the 48 technologies and analyzed in relation to patient characteristics and outcome. 49 Results: In spiked-in samples, the 3 technologies showed similar capability of recover epithelial 50 mammospheres, whereas, in a series of 19 clinical samples processed in parallel with the 51 CellSearch® and CellSieve filters (that allow the detection of both epithelial and non-epithelial 52 clusters), CTC-clusters were detected in 53% of samples with the CellSearch®, versus 79% and 84% 53 with the CellSieve, when considering only epithelial or both epithelial and non-epithelial clusters, 54 respectively. Abstract 39 55 Results: In spiked-in samples, the 3 technologies showed similar capability of recover epithelial 50 mammospheres, whereas, in a series of 19 clinical samples processed in parallel with the 51 CellSearch® and CellSieve filters (that allow the detection of both epithelial and non-epithelial 52 clusters), CTC-clusters were detected in 53% of samples with the CellSearch®, versus 79% and 84% 53 with the CellSieve, when considering only epithelial or both epithelial and non-epithelial clusters, 54 respectively. 55 Next, blood samples from 37 non-metastatic breast cancer (NMBC) and 23 MBC patients were 56 processed using ScreenCell® filters for attaining both unbiased enrichment and marker-independent 57 identification of clusters based on cytomorphological criteria. At baseline, CTC-clusters were 58 detected in 70% of NMBC cases and in 20% of MBC patients (median number= 2, range 0–20, 59 versus 0, range 0-15, P=0.0015). Among NMBC patients, clusters were slightly higher in women 60 with node-positive than node-negative status (0 versus 3, P=0.1110) and were more frequently 61 2 2 observed in women with luminal-like and triple-negative tumors than in patients with HER2-positive 62 disease (median CTC-cluster number =4, 5, and 0 for luminal-like, triple-negative, and HER2- 63 positive BC, respectively, P=0.0467). 64 Conclusions: We demonstrated that CTC-cluster detection can be improved by a marker-independent 65 enrichment and identification, and we reported that CTC-clusters are more frequently detected in 66 NMBC than in MBC patients, suggesting that dissemination of CTC-clusters is an early event in BC 67 natural history. 68 Conclusions: We demonstrated that CTC-cluster detection can be improved by a marker-independent 65 enrichment and identification, and we reported that CTC-clusters are more frequently detected in 66 NMBC than in MBC patients, suggesting that dissemination of CTC-clusters is an early event in BC 67 natural history. 68 Keywords: circulating tumor cell clusters, liquid biopsy, metastatic breast cancer, non-metastatic 69 breast cancer, circulating tumor microemboli, size-based enrichment 70 80 3 3 Background 81 Therefore, addressing the 103 4 presence of CTC-clusters in BC patients without clinically overt metastases holds promise to gain 104 important hints about the dissemination process. 105 presence of CTC-clusters in BC patients without clinically overt metastases holds promise to gain 104 important hints about the dissemination process. 105 presence of CTC-clusters in BC patients without clinically overt metastases holds promise to gain 104 important hints about the dissemination process. 105 However, this issue has not yet been addressed and, in BC, most studies evaluating the clinical 106 relevance of CTC-clusters have been limited to patients with metastatic or advanced disease [14-20]. 107 Overall, these studies suggest a direct association between detection of CTC-clusters and poor 108 clinical outcome, although the heterogeneous patient case series, technical issues in CTC-cluster 109 enumeration and variable definitions of CTC-clusters must be taken into account as possible 110 limitations and confounding factors. 111 However, this issue has not yet been addressed and, in BC, most studies evaluating the clinical 106 relevance of CTC-clusters have been limited to patients with metastatic or advanced disease [14-20]. 107 Overall, these studies suggest a direct association between detection of CTC-clusters and poor 108 clinical outcome, although the heterogeneous patient case series, technical issues in CTC-cluster 109 enumeration and variable definitions of CTC-clusters must be taken into account as possible 110 limitations and confounding factors. 111 Noteworthy, all the mentioned studies used the CellSearch® for CTC-cluster detection, which is 112 possibly not the ideal method for CTC-cluster identification. The CellSearch® is a platform 113 specifically developed for assuring high detection of single CTCs with epithelial features and for 114 attaining standardization of their enumeration [21]. No data are instead available on its performance 115 for CTC-cluster detection both in terms of recovery and of the integrity of isolated clusters. The 116 CellSearch® approach includes a CTC-enrichment step employing ferrofluid nanoparticles with 117 antibodies targeting EpCAM, which operates a selection in favor of clusters with exquisite epithelial 118 features and possibly excludes larger CTC-clusters [22], that could result into an underestimation in 119 CTC-cluster enumeration. Moreover, epithelial-to-mesenchymal transition (EMT) is recognized as an 120 important driver of tumor invasion and metastatic dissemination [23], and literature data supported an 121 increasing detection of mesenchymal markers in CTC-clusters compared to single CTCs in breast 122 cancer patients [24]. Background 81 Metastatic spreading is the main cause for death in patients diagnosed with cancer. This process is 82 promoted in its initial steps by cancer cell seeding from the primary tumor into the blood stream. 83 Accordingly, a large amount of data has been collected across different tumor types linking the 84 dissemination of circulating tumor cells (CTCs) with both poor prognosis and treatment 85 failure/resistance [1]. 86 Metastatic spreading is the main cause for death in patients diagnosed with cancer. This process is 82 promoted in its initial steps by cancer cell seeding from the primary tumor into the blood stream. 83 Accordingly, a large amount of data has been collected across different tumor types linking the 84 dissemination of circulating tumor cells (CTCs) with both poor prognosis and treatment 85 failure/resistance [1]. 86 Nonetheless, single CTCs are inefficient in sustaining metastatic dissemination as, to be able to 87 colonize new sites, they must overcome numerous obstacles such as avoid anoikis, escape 88 immunological control by circulating immune cells, resist to sharing stress due to fluid circulation, 89 resulting in the fact that most CTCs do not survive long in the circulation [2, 3]. Therefore, being 90 able to interact with other CTCs or with other cells by generating homo- or heterotypic CTC-clusters 91 appears a biologically reasonable solution for increasing the metastatic potential of CTCs once they 92 are facing the hostile blood environment. 93 Functional studies employing animal models and patient-derived data [4-7] definitely support a role 94 of CTC-clusters in tumor dissemination and metastasis formation in breast cancer (BC). Such studies 95 also offer hints on the biology of clusters revealing the mechanistic basis for their association with 96 poor outcome and suggesting possible targets for specific treatments aiming at interfere with CTC- 97 clusters formation and metastatic dissemination. 98 It is well known that metastatic dissemination occurs at early stages and is followed by a prolonged 99 dormant status of these early disseminated cells [8-10]. This observation is supported by data 100 demonstrating that enumeration of single CTCs predicts progression-free survival (PFS) and overall 101 survival (OS) also in non-metastatic breast cancer (NMBC) patients (women with no evidence for 102 distant metastases), both prior [11, 12] or after [13] breast surgery. Background 81 Thus, investigating the use of epitope-independent methods, compared to the 123 CellSearch®, for CTC-cluster detection is urgently needed to be able to fully appraise the actual 124 clinical value of CTC-cluster in BC. 125 To tackle technical issues in CTC-cluster enumeration we compared, in a series of spiked-in and 126 clinical samples, the number of CTC-clusters recovered using the CellSearch® platform and two 127 size-exclusion methods based on a short-time filtration that allows for the detection of both epithelial 128 5 and non-epithelial CTC-clusters. Thereafter, we implemented the recovery of CTC-clusters by 129 filtration in a prospective study involving patients with both NMBC and MBC to analyze CTC- 130 cluster detection with respect to patient and primary tumor features. 131 and non-epithelial CTC-clusters. Thereafter, we implemented the recovery of CTC-clusters by 129 filtration in a prospective study involving patients with both NMBC and MBC to analyze CTC- 130 cluster detection with respect to patient and primary tumor features. 131 Cell cultures and spiking experiments 134 The MCF7 breast cancer cell line was purchased from the American Type Culture Collection 135 (ATCC, Manassas, VA, USA) and cultured in DMEM/F-12 (Lonza, Slough, UK) medium 136 supplemented with 10% fetal bovine serum (Lonza). Mammospheres were derived from MCF7 cells 137 cultured as floating cells in MammoCult™ (StemCellTechnologies, Vancouver, Canada), a serum- 138 free culture medium, supplemented with Heparin Solution (StemCell Technologies) at final 139 concentration of 4 µg/mL, and Hydrocortisone (StemCell Technologies) at final concentration of 140 0.48 µg/mL. The cells were maintained in non-adherent condition (Corning® Ultra-Low Attachment 141 flask, Corning Inc., Corning, NY, USA) at 37 °C, in humidified 5% CO2 and 5% O2. Authentication 142 of cell lines by STR DNA profiling analysis was performed by the Genomic Core Facility at 143 Fondazione IRCCS Istituto Nazionale Tumori (INT). We adopt a Mycoplasma contamination testing 144 policy employing an ELISA approach (MycoAlert mycoplasma detection kit, Lonza) for regular 145 testing. 146 For the spiking experiments, either single MCF7 cells or single mammospheres were manually 147 captured under an inverted microscope using a p10 micropipette and directly spiked into phosphate- 148 buffered saline (PBS) supplemented with human serum albumin (HSA 3% w/v, to mimic protein 149 concentration of plasma), or into healthy donor blood collected in either CellSave Preservative Tubes 150 (Menarini Silicon Biosystems, Bologna, Italy) for CellSieve™ and CellSearch® processing, or in 151 6 6 K2EDTA BD Vacutainer tubes (BD, Franklin Lakes, NJ, USA) for ScreenCell® processing. Spiked- 152 in samples were processed following the same protocols used for clinical samples, described in 153 paragraphs 2.2 and 2.5. 154 Case series & blood sample collection timing 177 Women with a histologically confirmed diagnosis of stage II and III BC (NMBC) were recruited at 178 Fondazione IRCCS Istituto Nazionale dei Tumori (Milan, Italy) prior to start of neoadjuvant 179 treatment as for clinical practice, whereas women with stage IV BC (MBC) were recruited prior to 180 start of the first line of treatment. All patients provided written informed consent before undergoing 181 any procedures and the CTC study was approved by the INT Institutional Review Board and Ethics 182 Committee on February 19 2013. 183 Blood samples were longitudinally collected from patients with NMBC (i) before starting 184 neoadjuvant treatment, (ii) during treatment, (iii) at the end of treatment, and (iv) after surgery (from 185 3 to 27 weeks). Blood samples were collected from patients with MBC before starting the first line of 186 treatment. 187 Blood samples were longitudinally collected from patients with NMBC (i) before starting 184 neoadjuvant treatment, (ii) during treatment, (iii) at the end of treatment, and (iv) after surgery (from 185 3 to 27 weeks). Blood samples were collected from patients with MBC before starting the first line of 186 treatment. 187 Pathological complete response (pCR) was defined as the absence of cancer cells in the surgical 188 specimens of breast and lymph nodes. Partial response (PR) and stable disease (SD) referred to 189 clinical assessment of response to treatment according to the WHO criteria, hence a >50% tumor 190 shrinkage occurred for a PR, and >25% tumor increase for progressive disease (PD), whereas stable 191 disease was neither PR nor PD. 192 Pathological complete response (pCR) was defined as the absence of cancer cells in the surgical 188 specimens of breast and lymph nodes. Partial response (PR) and stable disease (SD) referred to 189 clinical assessment of response to treatment according to the WHO criteria, hence a >50% tumor 190 shrinkage occurred for a PR, and >25% tumor increase for progressive disease (PD), whereas stable 191 disease was neither PR nor PD. 192 Comparison of CellSearch® and CellSieve™ filters for CTC-cluster detection in clinical 155 samples 156 Blood samples (15 mL) were collected in CellSave Preservative Tubes (Menarini Silicon 157 Biosystems) from patients with MBC treated at the Robert H Lurie Comprehensive Cancer Center at 158 the Northwestern University (Chicago, IL, USA). All patients provided written informed consent to 159 participate in the study, which was approved by the institutional review board at the Robert H. Lurie 160 Comprehensive Cancer Center of Northwestern University (NUDB16Z01). Each sample was divided 161 into 2 aliquots (7.5 mL each) and processed in parallel with the CellSearch® (Menarini Silicon 162 Biosystems) and with CellSieve™ filters (Creatv MicroTech, Potomac, MD, USA) within 1 day from 163 collection. For CellSearch® processing, the CELLSEARCH® Circulating Tumor Cell Kit (Menarini 164 Silicon Biosystems) was used following the manufacturer’s instructions. Briefly, after 165 immunomagnetic enrichment based on EpCAM expression, enriched CTCs were stained with 166 fluorescently-labeled antibodies against cytokeratins (CK) (8, 18 and 19) and CD45 and with DAPI. 167 The number of CTC-clusters (groups of ≥ 2 CKpos/CD45neg cells) was evaluated using the 168 CELLTRACKS ANALYZER II® System (Menarini Silicon Biosystems) by a trained technician. For 169 CellSieve™ filters processing, the CellSieve™ Enumeration Kit (Creatv MicroTech) was used 170 following the manufacturer’s instructions. The blood samples were filtered through a microporous 171 membrane with pores of 7 µm diameter and subsequently stained with fluorescently-labeled 172 antibodies against CK (8, 18 and 19), CD45 and CD31 (an endothelial marker used to exclude 173 endothelial cell clusters) and with DAPI. The number of CTC-clusters (i.e. groups of ≥2 174 7 CKpos/CD45neg or ≥2 CKneg/CD45neg/CD31neg cells for CKpos and CKneg CTC-clusters, respectively) 175 was evaluated using a fluorescence microscope. 176 Patho-biological characterization of tumors 193 Hormone receptor status was evaluated according to the American Society of Clinical Oncology 194 guidelines [25]. HER2 status was considered negative when the immune-histochemical score was 0– 195 1, or 2+ with a negative chromogenic in situ hybridization result [26]. Ki-67 labeling index was 196 assessed by the MIB-1 monoclonal antibody by counting invasive cancer cells at the tumor 197 8 8 periphery, without focusing on hot-spots, as recommended by the International Ki-67 in Breast 198 Cancer Working Group [27]. 199 periphery, without focusing on hot-spots, as recommended by the International Ki-67 in Breast 198 Cancer Working Group [27]. 199 The evaluation of tumor-infiltrating lymphocytes (TILs) was performed in full-face hematoxylin and 200 eosin sections from surgical or bioptic sample, strictly adhering to the criteria proposed by the TILs 201 Working Group [28]. Briefly, all mononuclear cells (i.e. lymphocytes and plasma cells) in the 202 stromal compartment within the borders of the invasive tumor were evaluated and reported as a 203 percentage. TILs outside the tumour border, around in situ component (DCIS) and normal breast 204 tissue, as well as in areas of necrosis, were excluded from the scoring. 205 CTC-cluster enumeration by ScreenCell® filters 206 214 After rinsing with PBS, ISs were air-dried and stained with Hematoxyilin Solution S (Merck, 215 9 CTCs showing the criteria of malignancy: nuclear size ≥20 μm, nuclear-to-cytoplasmic ratio ≥0.75, 222 irregular nuclear contours and nuclear hyperchromatism. In case the cytoplasm edges were not 223 clearly visible inside the cluster (preventing nuclear-to-cytoplasmic ratio evaluation), malignancy 224 identification was mainly based on nuclei appearance: nuclei scattered irregularly through the cluster 225 and anisokaryosis (i.e. nuclei of variable sizes and shapes), in addition to nuclear size ≥20 μm and 226 irregular nuclear membrane. Detailed guidelines for ScreenCell filter interpretation are described 227 elsewhere [32]. Samples showing poor quality of cytology were excluded from the analysis. The total 228 number of CTC-clusters for each sample was obtained by summing the CTC-clusters identified in the 229 3 ISs (corresponding to 9 mL of blood). 230 CTCs showing the criteria of malignancy: nuclear size ≥20 μm, nuclear-to-cytoplasmic ratio ≥0.75, 222 irregular nuclear contours and nuclear hyperchromatism. In case the cytoplasm edges were not 223 clearly visible inside the cluster (preventing nuclear-to-cytoplasmic ratio evaluation), malignancy 224 identification was mainly based on nuclei appearance: nuclei scattered irregularly through the cluster 225 and anisokaryosis (i.e. nuclei of variable sizes and shapes), in addition to nuclear size ≥20 μm and 226 irregular nuclear membrane. Detailed guidelines for ScreenCell filter interpretation are described 227 elsewhere [32]. Samples showing poor quality of cytology were excluded from the analysis. The total 228 number of CTC-clusters for each sample was obtained by summing the CTC-clusters identified in the 229 3 ISs (corresponding to 9 mL of blood). 230 CTC-cluster enumeration by ScreenCell® filters 206 Peripheral blood samples (9 mL), collected into K2EDTA BD Vacutainer tubes (BD) using a 21G 207 needle, were stored at 4 °C in the dark and processed within 2.5 hours for CTC-cluster enrichment 208 using the ScreenCell® Cyto kit (ScreenCell, Sarcelles, France) [29] according to the manufacturer’s 209 instructions, with slight modifications with respect to what previously described [30, 31]. Briefly, 210 three aliquots of 3.0 mL of whole blood per sample were separately mixed with 4 mL of a proprietary 211 red blood cell lysis and fixation buffer (ScreenCell® FC2 filtration buffer) and incubated for 8 212 minutes at room temperature. Each aliquot was filtered to isolate CTC-clusters using ScreenCell® 213 Cyto isolation supports (ISs), consisting in a microporous membrane with pores of 6.5 µm diameter. 214 After rinsing with PBS, ISs were air-dried and stained with Hematoxyilin Solution S (Merck, 215 Darmstadt, Germany) for 1 min and Shandon Eosin Y Aqueous Solution (Thermo Fisher Scientific 216 Inc., Waltham, MA, USA) for 30 seconds, at room temperature; or with May Grünwald (Merck 217 Millipore, Burlington, MA, USA; incubation for 2.5 min followed by a second incubation for 2.5 min 218 in May Grünwald diluted 1:2 with water) and Giemsa (Merck Millipore; diluted 1:10 with water, 10 219 min incubation) at room temperature. The stained ISs were sent to ScreenCell for evaluation by a 220 certified pathologist according to published criteria [32]. CTC-clusters were defined as clusters of ≥2 221 Peripheral blood samples (9 mL), collected into K2EDTA BD Vacutainer tubes (BD) using a 21G 207 needle, were stored at 4 °C in the dark and processed within 2.5 hours for CTC-cluster enrichment 208 using the ScreenCell® Cyto kit (ScreenCell, Sarcelles, France) [29] according to the manufacturer’s 209 instructions, with slight modifications with respect to what previously described [30, 31]. Briefly, 210 three aliquots of 3.0 mL of whole blood per sample were separately mixed with 4 mL of a proprietary 211 red blood cell lysis and fixation buffer (ScreenCell® FC2 filtration buffer) and incubated for 8 212 minutes at room temperature. Each aliquot was filtered to isolate CTC-clusters using ScreenCell® 213 Cyto isolation supports (ISs), consisting in a microporous membrane with pores of 6.5 µm diameter. Statistical Analysis 231 Clinical and pathological variables were reported through descriptive analyses. Categorical variables 232 were reported as frequency distribution, whereas continuous variables were described according to 233 median and interquartile range (IQR). Differences in clusters distribution across subgroups of interest 234 were tested through Mann–Whitney U test. Pairwise comparison between CellSearch® and 235 CellSieve™ technologies, and across different timepoints during neoadjuvant therapy were 236 performed though Wilcoxon sign-rank test. All reported P-values are two-sided. 237 Clinical and pathological variables were reported through descriptive analyses. Categorical variables 232 were reported as frequency distribution, whereas continuous variables were described according to 233 median and interquartile range (IQR). Differences in clusters distribution across subgroups of interest 234 were tested through Mann–Whitney U test. Pairwise comparison between CellSearch® and 235 CellSieve™ technologies, and across different timepoints during neoadjuvant therapy were 236 performed though Wilcoxon sign-rank test. All reported P-values are two-sided. 237 Statistical analysis was conducted using the StataCorp 2016 Stata Statistical Software: Release 15.1 238 (College Station, Texas, USA), R (The R foundation for Statistical Computing. version 3.3.1) (2016- 239 06-21) and JMP (SAS Institute, version 15). 240 Statistical analysis was conducted using the StataCorp 2016 Stata Statistical Software: Release 15.1 238 (College Station, Texas, USA), R (The R foundation for Statistical Computing. version 3.3.1) (2016- 239 06-21) and JMP (SAS Institute, version 15). 240 243 10 Technical validation of approaches used for CTC-cluster detection 246 To explore technical limitations of standard (CellSearch®) and filtration-based methods for 247 CTC-cluster detection, spiking experiments were performed comparing size-exclusion approaches 248 with the CellSearch® method (the currently most frequently used method in CTC-cluster studies). In 249 particular, we compared 3 technologies: the CellSearch®, the CellSieve™ filters and the 250 ScreenCell® filters. The latter two are very similar for the enrichment strategy (based on short-time 251 filtration through a membrane with pores of 7 and 6.5 µm, respectively), but differ for the criteria 252 employed for the identification of tumor cells. CellSieve™ filters include an identification based on 253 marker expression similar to the one of the CellSearch® (CKpos and CD45neg cells), whereas 254 ScreenCell® filters’ identification is based on cytomorphological evaluation. 255 Mammospheres derived from the MCF7 breast cancer cell line were used as a surrogate of 256 CTC-clusters. For each technology, 10 mammospheres were spiked into healthy donor blood samples 257 (n = 8), and subsequently processed for CTC-cluster enrichment. For ScreenCell® only, 2 spiking 258 experiments were performed using PBS supplemented with HSA, instead of blood; this was done to 259 test the stress associated with the filtration process itself, since, for this technology only, fresh blood 260 is used for the spiking step and the presence of active immune cells from the donor might have an 261 impact on mammospheres (for CellSearch® and CellSieve™ the blood is instead collected in 262 CellSave tubes containing a preservative which fix blood cells). The number of recovered 263 mammospheres for each of the 10 spiking experiments are reported in Table 1. 264 11 Table 1. Mammospheres identified in spiking experiments, using different detection methods 267 Sample ID Enrichment technology N. of recovered mammospheres 1 CellSearch® 7 2 8 3 6 4 CellSieve™ 7 5 8 6 6 7 ScreenCell® 6 8 10 9* 10 10* 11 For samples 9 and 10, mammospheres were spiked into PBS supplemented with HSA, instead of 268 blood. 269 270 267 Table 1. Mammospheres identified in spiking experiments, using different detection methods 267 Sample ID Enrichment technology N. of recovered mammospheres 1 CellSearch® 7 2 8 3 6 4 CellSieve™ 7 5 8 6 6 7 ScreenCell® 6 8 10 9 10 10* 11 For samples 9 and 10, mammospheres were spiked into PBS supplemented with HSA, instead of 268 blood. 269 270 Table 1. Technical validation of approaches used for CTC-cluster detection 246 In that respect, 19 blood samples collected from 16 patients with MBC were processed 290 in parallel with CellSearch® and CellSieve™ filters (Fig. 1A). For this analysis, CellSieve™ was 291 used as the representative among the 2 filtration methods, since its enrichment strategy is the same of 292 ScreenCell filters (based on size), but its identification criteria are based on the detection of epithelial 293 markers, and therefore allow for the distinction between epithelial and non-epithelial clusters (not 294 possible with ScreenCell® filters). 295 Once assessed that the ability of enriching clusters for the 3 technologies was similar, we next aimed 286 at evaluating whether phenotypic heterogeneity of CTC-clusters in clinical samples (i.e. the presence 287 of both epithelial and non-epithelial clusters) could have an impact on CTC-cluster detection by the 288 epithelial marker-based CellSearch® platform, compared to the marker-independent and size-based 289 approaches. In that respect, 19 blood samples collected from 16 patients with MBC were processed 290 in parallel with CellSearch® and CellSieve™ filters (Fig. 1A). For this analysis, CellSieve™ was 291 used as the representative among the 2 filtration methods, since its enrichment strategy is the same of 292 ScreenCell filters (based on size), but its identification criteria are based on the detection of epithelial 293 markers, and therefore allow for the distinction between epithelial and non-epithelial clusters (not 294 possible with ScreenCell® filters). 295 Blood samples were collected from clinically selected patients with highly aggressive disease and 296 during disease progression to increase the probability of CTC-cluster presence (in Additional file 1, 297 patients’ clinico-pathological characteristics are reported). For samples processed with the 298 CellSearch®, only CTC-clusters expressing CK (CKpos CTC-clusters, defined as groups of 2 or more 299 cells showing CKpos and CD45neg staining, Fig. 1B) could be detected, whereas for samples processed 300 with CellSieve™ filters it was possible to identify both CKpos and CKneg clusters (Fig. 1C and 1D, 301 respectively). CKneg clusters were defined as groups of 2 or more cells showing a CKneg, CD45neg and 302 CD31neg staining (the latter marker allowing for the exclusion of endothelial cell clusters). CD31 303 expression was unexpectedly observed also in a few CKpos CTC-clusters (Additional file 2). These 304 clusters, being CKpos and CD45neg were included in the analysis. 305 We detected ≥1 CKpos CTC-clusters in 10 samples by using the CellSearch® and in 15 samples by 306 using CellSieve™ filters (Additional file 3). Technical validation of approaches used for CTC-cluster detection 246 Mammospheres identified in spiking experiments, using different detection methods 7 spheres identified in spiking experiments, using different detection methods For samples 9 and 10, mammospheres were spiked into PBS supplemented with HSA, instead of 268 blood. 269 270 All three technologies showed similar recovery ranging from 60% to 100%. The impossibility of 271 recovering 100% of the mammospheres in each sample suggests that a partial dissociation of the 272 mammospheres occurred, as also supported by the presence of single tumor cells and fragments of 273 mammospheres in the samples. However, disruption was not specifically induced by filtration, in fac 274 all the samples with a 100% recovery were processed with filters. 275 *For samples 9 and 10, mammospheres were spiked into PBS supplemented with HSA, instead of 268 blood. 269 270 All three technologies showed similar recovery ranging from 60% to 100%. The impossibility of 271 recovering 100% of the mammospheres in each sample suggests that a partial dissociation of the 272 mammospheres occurred, as also supported by the presence of single tumor cells and fragments of 273 mammospheres in the samples. However, disruption was not specifically induced by filtration, in fact 274 all the samples with a 100% recovery were processed with filters. 275 Another possible concern about using filtration devices for CTC-cluster identification, is the 276 possibility of the formation of aggregates on the filtration membrane during the processing, resulting 277 in the identification of fictitious CTC-clusters. To exclude this possibility, we spiked single MCF7 278 cells in 3 healthy donor blood samples (n = 30 MCF7 cells per sample). The samples were processed 279 with CellSieve™ filters and with ScreenCell® filters (2 and 1 sample, respectively). We observed the 280 presence of only 1 aggregate of 2 tumor cells, on one CellSieve™ filter, indicating that filtration does 281 not induce the formation of artifactual clusters. 282 12 CTC-cluster detection in clinical samples using an epithelial-based and a size-based approach 285 Once assessed that the ability of enriching clusters for the 3 technologies was similar, we next aimed 286 at evaluating whether phenotypic heterogeneity of CTC-clusters in clinical samples (i.e. the presence 287 of both epithelial and non-epithelial clusters) could have an impact on CTC-cluster detection by the 288 epithelial marker-based CellSearch® platform, compared to the marker-independent and size-based 289 approaches. Technical validation of approaches used for CTC-cluster detection 246 Moreover, in the samples processed by filtration, CKneg 307 clusters were observed in 12 out of 18 evaluable samples, in 1 case alone and in 11 cases together 308 We detected ≥1 CKpos CTC-clusters in 10 samples by using the CellSearch® and in 15 samples by 306 using CellSieve™ filters (Additional file 3). Moreover, in the samples processed by filtration, CKneg 307 clusters were observed in 12 out of 18 evaluable samples, in 1 case alone and in 11 cases together 308 13 with CKpos CTC-clusters. Overall, by considering 1 cluster per sample as positivity threshold, the 309 positivity rate increased by using the size-based approach, going from 53 % (using the CellSearch®) 310 to 79 % considering only CKpos and 84 % considering both CKpos and CKneg clusters identified with 311 CellSieve™ filters (Fig. 1E). Moreover, the absolute numbers of detected clusters were higher in 312 samples processed with CellSieve™ filters than with the CellSearch® (Fig. 1F; Additional file 3). In 313 samples processed with the CellSearch®, a median of 1 CKpos CTC-cluster (interquantile range, 314 IQR = 0-2; range 0-108) was identified, compared to a median of 3 CKpos CTC-clusters (IQR 1-6; 315 range 0-112) for samples processed with CellSieve™ filters (P = 0.0293). The increase in cluster 316 counts for samples processed with CellSieve™ filters was even higher when considering CKpos and 317 CKneg clusters together (median = 7, IQR 1-11; range 0-112, P = 0.0038). 318 with CKpos CTC-clusters. Overall, by considering 1 cluster per sample as positivity threshold, the 309 positivity rate increased by using the size-based approach, going from 53 % (using the CellSearch®) 310 to 79 % considering only CKpos and 84 % considering both CKpos and CKneg clusters identified with 311 CellSieve™ filters (Fig. 1E). Moreover, the absolute numbers of detected clusters were higher in 312 samples processed with CellSieve™ filters than with the CellSearch® (Fig. 1F; Additional file 3). In 313 samples processed with the CellSearch®, a median of 1 CKpos CTC-cluster (interquantile range, 314 IQR = 0-2; range 0-108) was identified, compared to a median of 3 CKpos CTC-clusters (IQR 1-6; 315 range 0-112) for samples processed with CellSieve™ filters (P = 0.0293). The increase in cluster 316 counts for samples processed with CellSieve™ filters was even higher when considering CKpos and 317 CKneg clusters together (median = 7, IQR 1-11; range 0-112, P = 0.0038). Technical validation of approaches used for CTC-cluster detection 246 318 These results suggest that by using a size-based and marker-independent approach it is possible to 319 detect a higher number of clusters, allowing to identify them also in patients considered CTC-cluster 320 negative by the CellSearch®. However, the observed phenotypic heterogeneity of clusters in BC 321 patient samples, and in particular the presence of CKneg clusters, highlighted an important limitation 322 of CellSieve™ technology, which was able to enrich this type of clusters, but did not allow to 323 reliably assess their malignancy (since they were only DAPIpos). On the other hand, ScreenCell® 324 technology had the same ability of enriching CKneg clusters (since it is size-based as well), but its 325 identification was based on cytomorphological evaluation and was therefore not dependent on the 326 expression of any specific tumor markers. We therefore decided to use ScreenCell® filters to 327 investigate the presence of CTC-clusters in both MBC and NMBC patients. 328 Patient characteristics 330 Between June 2014 and December 2015 a total of 37 and 23 patients with NMBC and MBC 331 undergoing systemic treatment were enrolled in the study. Main clinical and pathological features are 332 reported in Tables 2 and 3 (NMBC and MBC patients, respectively). 333 14 Table 2. Clinico-pathological characteristics of NMBC patients and CTC-clusters 334 Table 2. Clinico-pathological characteristics of NMBC patients and CTC-clusters N % median CTC- clusters (range) p CTC-cluster + (%) p Age • <50 20 54.1 2.5 (0-20) 0.889 15 (75%) 0.719 • ≥50 17 45.9 2 (0-20) 11 (65%) Tumor size • T1-T2 21 56.8 4 (0-20) 0.180 16 (76%) 0.475 • ≥T3 16 43.2 1 (0-15) 10 (63%) Nodal status • N0 8 21.6 0 (0-12) 0.273 3 (37.5) 0.123 • N1 21 56.8 3 (0-20) 17 (81.1) • ≥N2 8 21.6 3 (0-20) 6 (75%) Histology • NST 35 94.6 2 (0-20) 0.322 15(68%) >0.99 • Lobular 2 5.4 3 (0-15) 11(73%) Grade • 2 10 27.0 2 (0-15) 0.918 7 (70%) >0.99 • 3 22 59.5 1.5 (0-20) 15 (68.2%) • Missing 5 13.5 Ki67 • <20 4 10.8 1.5 (0-12) >0.10 2 (50%) 0.570 • ≥ 20 32 86.5 2 (0-20) 23 (72%) • Missing 1 2.7 - - Subtype • HER2-positive 11 29.7 0 (0-8) 0.047 5 (45%) 0.111 • Triple negative 11 29.7 5 (0-20) 9 (82%) • Luminal-like 15 40.5 4 (0-20) 12 (80%) Type of neoadjuvant chemotherapy • Anthra/Taxane 32 86.5 2.5 (0-20) 0.984 22 (69%) 0.609 • CarboPt-based 5 13.5 1 (0-20) 4 (80%) 15 Table 3. Clinico-pathological characteristics of MBC patients 337 338 Table 3. Patient characteristics 330 Clinico-pathological characteristics of MBC patients 337 338 337 338 N % Age • <50 5 21.7 • ≥50 18 78.3 Histology • Ductal 15 65.2 • Lobular 2 8.7 • Other 6 26.1 Disease type at screening • Visceral 6 26.1 • Nonvisceral 12 52.2 • Missing 3 13.0 Hormone receptor status • ER -positive, PgR positive or both 18 78.3 • ER-negative and PgR-negative 5 21.7 HER2 status • Positive 1 4.3 • Negative 22 95.7 Metastatic disease at diagnosis • No 15 65.2 • Yes 8 34.8 Prior chemotherapy for metastatic disease • No 20 87.0 • Yes 3 13.0 N % Age • <50 5 21.7 • ≥50 18 78.3 Histology • Ductal 15 65.2 • Lobular 2 8.7 • Other 6 26.1 Disease type at screening • Visceral 6 26.1 • Nonvisceral 12 52.2 • Missing 3 13.0 Hormone receptor status • ER -positive, PgR positive or both 18 78.3 • ER-negative and PgR-negative 5 21.7 HER2 status • Positive 1 4.3 • Negative 22 95.7 Metastatic disease at diagnosis • No 15 65.2 • Yes 8 34.8 Prior chemotherapy for metastatic disease • No 20 87.0 • Yes 3 13.0 339 The median age of NMBC patients treated with neoadjuvant chemotherapy (NAC) was 49 years 340 The median age of NMBC patients treated with neoadjuvant chemotherapy (NAC) was 49 years 340 (range 26-84). At diagnosis, tumor size was 2-5 cm (cT2) in 20 patients (54 %), and > 5 cm (cT3-4) 341 in 16 patients (43 %). Clinical nodal status was positive (cN1-3) in 29 cases (78 %). No patients with 342 stage I BC were enrolled. Histological grade 3 was reported in 22 evaluable patients (60 %). Among 343 the 36 evaluable patients, the median Ki67 value was 40 %, with values ranging from 10 % to 90 %. 344 16 Thirty-two patients (86 %) had primary tumors with a Ki67 staining ≥ 20 %. Nine patients (24.3 %) 345 reached a pathological complete response (pCR). 346 Thirty-two patients (86 %) had primary tumors with a Ki67 staining ≥ 20 %. Nine patients (24.3 %) 345 reached a pathological complete response (pCR). 346 The median age of MBC patients was 68 years (range 29-84). The most common histological type 347 was invasive ductal carcinoma (65 % of cases). Patient characteristics 330 Of the 23 patients included in the study, 6 (26 %) had 348 visceral and 12 (52 %) had non-visceral involvement. Six patients (26 %) presented with de novo 349 metastases. All patients, except three, had received no prior systemic treatment for metastatic disease. 350 CTC-clusters in patients with metastatic and non-metastatic breast cancer 351 CTC-clusters in patients with metastatic and non-metastatic breast cancer 351 To investigate the presence of CTC-clusters in our cohort of patients with MBC and NMBC, blood 352 samples collected before starting systemic treatment underwent CTC-cluster enrichment by filtration, 353 followed by a marker-independent CTC-cluster identification based on cytomorphological criteria 354 using ScreenCell® filters (Fig. 2A). This simplified identification strategy requires only H&E 355 staining rather than immunofluorescence, and it gives reliable results regarding cell malignancy, 356 independently from the expression of specific markers. At baseline, in NMBC patients, 1 or more 357 CTC-clusters were detected in 26/37 cases (70 %), with a median of 2 clusters per sample (range 358 0-20) (Fig. 2B). Among the 23 baseline samples collected from MBC patients, 3 samples were from 359 pre-treated patients and one was not evaluable for CTC-cluster identification; CTC-clusters were 360 detected in 4 of the 19 remaining samples (21 %), with a median of 0 CTC-clusters per sample (range 361 0-15). CTC-clusters were therefore more frequent and more abundant in patients with NMBC than 362 MBC (P = 0.0015). In particular, patients with stage II BC showed a higher CTC-cluster count than 363 patients with stage III and IV BC (Additional file 4A). Among patients with NMBC, a slightly higher 364 number of CTC-clusters was detected in patients with node-positive status (Fig. 2C), although this 365 difference was not statistically significant (median CTC-cluster number = 0 vs. 3 for node-negative 366 vs. node-positive patients, P = 0.1110). CTC-clusters were more frequently observed in patients with 367 luminal-like and triple negative BC than in patients with HER2-positive disease (median CTC-cluster 368 To investigate the presence of CTC-clusters in our cohort of patients with MBC and NMBC, blood 352 samples collected before starting systemic treatment underwent CTC-cluster enrichment by filtration, 353 followed by a marker-independent CTC-cluster identification based on cytomorphological criteria 354 using ScreenCell® filters (Fig. 2A). This simplified identification strategy requires only H&E 355 staining rather than immunofluorescence, and it gives reliable results regarding cell malignancy, 356 independently from the expression of specific markers. At baseline, in NMBC patients, 1 or more 357 CTC-clusters were detected in 26/37 cases (70 %), with a median of 2 clusters per sample (range 358 0-20) (Fig. 2B). CTC-clusters in patients with metastatic and non-metastatic breast cancer 351 Among the 23 baseline samples collected from MBC patients, 3 samples were from 359 pre-treated patients and one was not evaluable for CTC-cluster identification; CTC-clusters were 360 detected in 4 of the 19 remaining samples (21 %), with a median of 0 CTC-clusters per sample (range 361 0-15). CTC-clusters were therefore more frequent and more abundant in patients with NMBC than 362 MBC (P = 0.0015). In particular, patients with stage II BC showed a higher CTC-cluster count than 363 patients with stage III and IV BC (Additional file 4A). Among patients with NMBC, a slightly higher 364 number of CTC-clusters was detected in patients with node-positive status (Fig. 2C), although this 365 difference was not statistically significant (median CTC-cluster number = 0 vs. 3 for node-negative 366 vs. node-positive patients, P = 0.1110). CTC-clusters were more frequently observed in patients with 367 luminal-like and triple negative BC than in patients with HER2-positive disease (median CTC-cluster 368 17 number = 4, 5, and 0 for luminal-like, triple-negative, and HER2-positive BC respectively, P = 369 0.0467) (Fig. 2D). For 25 patients for whom a primary tumor tissue sample was available, the 370 presence of CTC-clusters was analyzed with respect to the presence of tumor-infiltrating 371 lymphocytes (TILs) at the primary tumor site but no difference in CTC-cluster counts was observed 372 between patients presenting a high or low level of TILs (median CTC-cluster number = 3 vs. 2 for 373 patients with < 12 % vs. ≥ 12 % TILs, P = 0.5392) (Additional file 4B). 374 These results indicate that CTC-clusters are present in early stages in BC patients and are more 375 frequent than in MBC patients. Among NMBC patients, CTC-clusters are more abundant in the 376 blood of patients with HER2-negative disease. 377 These results indicate that CTC-clusters are present in early stages in BC patients and are more 375 frequent than in MBC patients. Among NMBC patients, CTC-clusters are more abundant in the 376 blood of patients with HER2-negative disease. 377 Longitudinal evaluation of CTC-clusters during neoadjuvant therapy 378 Longitudinal evaluation of CTC-clusters during neoadjuvant therapy 378 At the same date, 4 out of 19 evaluable MBC patients had died, notably the negative 398 predictive value of clusters at baseline in this case was as high as 86%, but the data is merely 399 explorative due to the small sample size. 400 associated with pCR (Additional file 5A). However, patients without clusters at baseline reported a 392 numerically higher pCR rate as compared with those presenting with clusters, 27% versus 23%, 393 respectively. Moreover, after surgery, a significantly lower number of clusters was observed in 394 patients with pathological complete or partial response versus stable disease (P = 0.0208) (Additional 395 file 5B). As of May 15, 2020, a total of 10 out of 37 NMBC patients relapsed. No difference in 396 baseline or post treatment distribution of clusters was reported among patients with or without a 397 relapse. At the same date, 4 out of 19 evaluable MBC patients had died, notably the negative 398 predictive value of clusters at baseline in this case was as high as 86%, but the data is merely 399 explorative due to the small sample size. 400 We present two examples of patients who responded to NAC but did not achieve pCR, illustrating 401 the cluster’s dynamics during treatment. 402 We present two examples of patients who responded to NAC but did not achieve pCR, illustrating 401 the cluster’s dynamics during treatment. 402 Patient A (Fig. 3B) was diagnosed with a 40 mm ductal carcinoma of the right breast, G3 ER, PgR 403 and HER2 negative, 90% Ki67. Bone scan and liver ultrasound were negative for distant 404 involvement. She was further staged with positron emission tomography (PET) scan that confirmed a 405 breast primary lesion with a standardized uptake value (SUV) of 22.5 and no loco-regional 406 involvement. The patient was therefore enrolled in a NAC clinical trial and received 4 cycles of 407 Doxorubicin 60 mg/m2 together with Paclitaxel 200 mg/m2 q21. No clusters were detectable at 408 baseline. The first PET evaluation showed a dramatic drop in metabolic activity (SUV 3.7), with 5 409 clusters detectable in the peripheral blood. Eribulin 1.23 mg/m2 was then started and 4 cycles were 410 administered with a 1, 8, q21 schedule. The subsequent PET scan showed further metabolic response 411 with a 3.7 SUV, while an increase in clusters was observed (35 clusters). Longitudinal evaluation of CTC-clusters during neoadjuvant therapy 378 Longitudinal evaluation of CTC-clusters during neoadjuvant therapy 378 To further investigate the clinical relevance of CTC-clusters in NMBC patients, longitudinal blood 379 samples collected at baseline (N = 37), during (N = 30), at the end (N = 14) of NAC and after surgery 380 (N = 18) were analyzed (Fig.3A). The median number of detected CTC-clusters at baseline was 2 381 (range 0-20), during treatment (DT) was 1 (range 0-97), and at the end of treatment (EOT) was 3 382 (range 0-116). Thus, CTC-clusters did not decrease during NAC, but instead increased in some 383 patients. Overall, no significant differences were observed in DT and EOT with respect to baseline. 384 On the other hand, a significant decrease was observed from DT to surgery (P = 0.0448) and EOT to 385 surgery (P = 0.0208). Only a slight decrease was instead observed between baseline and surgery (P = 386 0.0678). The median number of CTC-clusters after surgery was 0 (range 0-20). 387 At baseline, numerically less clusters were observed in NAC-responders, i.e. patients with complete 388 disappearance or a reduction of primary tumor volume of at least 50% after NAC, as compared to 389 non-responders, i.e. patients with stable disease after NAC: 1 cluster (range 0-20) versus 4 clusters 390 (range 0-12), respectively (P = 0.58). The presence of CTC-clusters at baseline was not significantly 391 At baseline, numerically less clusters were observed in NAC-responders, i.e. patients with complete 388 disappearance or a reduction of primary tumor volume of at least 50% after NAC, as compared to 389 non-responders, i.e. patients with stable disease after NAC: 1 cluster (range 0-20) versus 4 clusters 390 (range 0-12), respectively (P = 0.58). The presence of CTC-clusters at baseline was not significantly 391 18 associated with pCR (Additional file 5A). However, patients without clusters at baseline reported a 392 numerically higher pCR rate as compared with those presenting with clusters, 27% versus 23%, 393 respectively. Moreover, after surgery, a significantly lower number of clusters was observed in 394 patients with pathological complete or partial response versus stable disease (P = 0.0208) (Additional 395 file 5B). As of May 15, 2020, a total of 10 out of 37 NMBC patients relapsed. No difference in 396 baseline or post treatment distribution of clusters was reported among patients with or without a 397 relapse. Longitudinal evaluation of CTC-clusters during neoadjuvant therapy 378 She then underwent 412 quadrantectomy with 17 mm residual disease and absence of nodal involvement (ypT1c, N0). ER 413 was 2%, PgR and HER2 were negative, Ki67 was confirmed at 90%. Filter based enumeration after 414 surgery showed a complete clearance of detectable clusters. The patient was then started on adjuvant 415 19 CMF (Cyclophosphamide 600 mg/m2, methotrexate 40 mg/m2 and 5-Fluorouracil 600 mg/m2) but 416 died 7 months after surgery for noncancer-related causes without any detectable local or distant 417 relapse. 418 CMF (Cyclophosphamide 600 mg/m2, methotrexate 40 mg/m2 and 5-Fluorouracil 600 mg/m2) but 416 died 7 months after surgery for noncancer-related causes without any detectable local or distant 417 relapse. 418 Patient B was diagnosed with a screening-detected lobular carcinoma of the left breast, G2, ER 20%, 419 PgR 10%, HER2 negative and Ki67 10%. The baseline breast magnetic resonance (MRI) showed a 420 multifocal primary tumor with a 38 mm main lesion and a 3 mm satellite lesion, while distant 421 metastases were excluded through PET scan. Baseline clusters enumeration was 3. A Doxorubicin 60 422 mg/m2 and Paclitaxel 200 mg/m2 q21 based NAC was started. Breast MRI after 4 cycles showed a 423 partial regression. Clusters were not detectable. CMF was administered for 4 cycles. While the breast 424 MRI showed further radiological response, 116 clusters were detected in the peripheral blood. The 425 patient underwent quadrantectomy with 3 mm residual disease and 2 metastatic lymph nodes out of 7 426 analyzed (ypT1a N1a). ER was 90 %, PgR and HER2 were negative, Ki-67 was 5 %. After bone scan 427 restaging, she was started on adjuvant Letrozole 2.5mg, which is still ongoing without evidence of 428 distant or local recurrence. 429 Discussion 431 In the current study, we have challenged the most frequently used technical approach for CTC-cluster 432 detection in BC, the CellSearch®, by comparing it with methods based on size exclusion. We report 433 that filtration allowed to detect a higher number of clusters in the blood of BC patients. Moreover, by 434 using a filtration-based approach to analyze blood samples prospectively collected from NMBC and 435 MBC patients, we observed that CTC-clusters were more frequently detected in NMBC than in MBC 436 patients, and that molecular subtypes affected their presence in NMBC. Finally, the presence of 437 In the current study, we have challenged the most frequently used technical approach for CTC-cluster 432 detection in BC, the CellSearch®, by comparing it with methods based on size exclusion. We report 433 that filtration allowed to detect a higher number of clusters in the blood of BC patients. Moreover, by 434 using a filtration-based approach to analyze blood samples prospectively collected from NMBC and 435 MBC patients, we observed that CTC-clusters were more frequently detected in NMBC than in MBC 436 patients, and that molecular subtypes affected their presence in NMBC. Finally, the presence of 437 20 20 clusters before starting neoadjuvant treatment did not associate with pCR and their numbers 438 increased during treatment, but dropped after surgery. 439 clusters before starting neoadjuvant treatment did not associate with pCR and their numbers 438 increased during treatment, but dropped after surgery. 439 clusters before starting neoadjuvant treatment did not associate with pCR and their numbers 438 increased during treatment, but dropped after surgery. 439 To the best of our knowledge, this is the first study specifically comparing CTC-cluster detection by 440 CellSearch® and by a validated filtration-based technique [33], in patients with BC. Such a 441 comparison has instead been performed in small-cell lung cancer patients, by using in parallel the 442 CellSearch® and the ISET filtration approach, showing similar results [22]. Indeed in lung cancer 443 patients no clusters were detected with the CellSearch®, whereas they were found in 50% of samples 444 from stage IIIB/IV patients processed with the ISET. The findings were explained by the authors as a 445 possible failure of the immune-magnetic enrichment step in the CellSearch® protocol to capture 446 large size clusters. Discussion 431 However, additional considerations can be made regarding the increased 447 CTC-cluster detection attained by using filtration, both in our study and in that by Krebs and 448 colleagues. Strong positivity for mesenchymal, with concomitant weak positivity for epithelial 449 markers, has been reported for CTC-clusters isolated from patients with advanced breast cancer [24]. 450 Thus, an increase in cluster detection is not surprising when using methods that do not relay on the 451 expression of epithelial markers, and which are not limited to the detection of epithelial clusters only. 452 Conversely, the observed increased detection of epithelial clusters (CKpos) is an unexpected finding. 453 A possible explanation is that CKpos CTC-clusters can also include cells undergoing EMT and 454 therefore expressing a mixed phenotype rather than a frankly epithelial one. Since the CellSearch® 455 detects only CTC-clusters expressing both EpCAM and CK, but EpCAM expression is lost early 456 during EMT [23, 34] the CellSearch could miss CKpos CTC-clusters that are going through the 457 EMT. This hypothesis could not be verified in the present study since the expression of mesenchymal 458 markers was not assessed. However, it is supported by the results of the spiking experiments showing 459 that when using frankly epithelial mammospheres (expressing both EpCAM and CK), CellSearch 460 yielded comparable recovery rates as filtration devices. 461 increased during treatment, but dropped after surgery. 439 To the best of our knowledge, this is the first study specifically comparing CTC-cluster detection by 440 CellSearch® and by a validated filtration-based technique [33], in patients with BC. Such a 441 comparison has instead been performed in small-cell lung cancer patients, by using in parallel the 442 CellSearch® and the ISET filtration approach, showing similar results [22]. Indeed in lung cancer 443 patients no clusters were detected with the CellSearch®, whereas they were found in 50% of samples 444 from stage IIIB/IV patients processed with the ISET. The findings were explained by the authors as a 445 possible failure of the immune-magnetic enrichment step in the CellSearch® protocol to capture 446 large size clusters. However, additional considerations can be made regarding the increased 447 CTC-cluster detection attained by using filtration, both in our study and in that by Krebs and 448 colleagues. Strong positivity for mesenchymal, with concomitant weak positivity for epithelial 449 markers, has been reported for CTC-clusters isolated from patients with advanced breast cancer [24]. Discussion 431 450 Thus, an increase in cluster detection is not surprising when using methods that do not relay on the 451 expression of epithelial markers, and which are not limited to the detection of epithelial clusters only. 452 Conversely, the observed increased detection of epithelial clusters (CKpos) is an unexpected finding. 453 A possible explanation is that CKpos CTC-clusters can also include cells undergoing EMT and 454 therefore expressing a mixed phenotype rather than a frankly epithelial one. Since the CellSearch® 455 detects only CTC-clusters expressing both EpCAM and CK, but EpCAM expression is lost early 456 during EMT [23, 34] the CellSearch could miss CKpos CTC-clusters that are going through the 457 EMT. This hypothesis could not be verified in the present study since the expression of mesenchymal 458 markers was not assessed. However, it is supported by the results of the spiking experiments showing 459 that when using frankly epithelial mammospheres (expressing both EpCAM and CK), CellSearch 460 yielded comparable recovery rates as filtration devices. 461 To the best of our knowledge, this is the first study specifically comparing CTC-cluster detection by 440 CellSearch® and by a validated filtration-based technique [33], in patients with BC. Such a 441 21 21 21 Besides filters, other marker-independent technologies such as the HBCTC-Chip [4, 24], the Cluster- 462 Chip [35] and the Parsortix™ [6], have been employed for CTC-cluster studies, but mainly focusing 463 on functional aspects rather than on pure translational purposes. In fact, despite a number of studies 464 have described new technical tools specifically dedicated to CTC-cluster detection [for a review see 465 36], none of these innovative methods is widely available to clinical research centers. In this context, 466 simpler technologies, as those based on size-exclusion would represent a more affordable approach, 467 easily transferable to clinical studies that might help in elucidating the role of CTC-clusters in 468 different clinical contexts. 469 Besides filters, other marker-independent technologies such as the HBCTC-Chip [4, 24], the Cluster- 462 Chip [35] and the Parsortix™ [6], have been employed for CTC-cluster studies, but mainly focusing 463 on functional aspects rather than on pure translational purposes. In fact, despite a number of studies 464 have described new technical tools specifically dedicated to CTC-cluster detection [for a review see 465 36], none of these innovative methods is widely available to clinical research centers. Discussion 431 Moreover, we have noticed that luminal-like tumors release 490 high number of clusters, a finding possibly linked to their late relapse-pattern and to a more efficient 491 promotion of dormancy within the clusters from patients with ER+ tumors [41]. Overall, this 492 suggests that clusters should be studied in molecularly homogeneous populations, although this could 493 not be done in this study, due to the limited number of patients. 494 Nevertheless, many questions on clinical and biological aspects still remain to be answered. We 486 observed that molecular subtypes affect the prevalence of CTC-clusters. In particular CTC-clusters 487 were found to be significantly more frequent in women bearing HER2-negative tumors, a result that 488 may appear as contra-intuitive since HER2-positive tumors are more aggressive and are frequently 489 associated with stemness markers [40]. Moreover, we have noticed that luminal-like tumors release 490 high number of clusters, a finding possibly linked to their late relapse-pattern and to a more efficient 491 promotion of dormancy within the clusters from patients with ER+ tumors [41]. Overall, this 492 suggests that clusters should be studied in molecularly homogeneous populations, although this could 493 not be done in this study, due to the limited number of patients. 494 In our cohort of NMBC patients, the detection of clusters did not correlate with the likelihood of 495 achieving pCR, a finding already reported in the literature for CTCs [12, 38]. Moreover, during the 496 course of treatment a trend towards an increase in CTC-clusters rather than a decrease was observed, 497 as also described in another study using ScreenCell® filters [38]. Indeed, only after surgery we 498 actually observed a decrease in the number of clusters. Although a significantly higher number of 499 clusters persisting after surgery was detected in patients with a pathologically non-responding disease 500 (median 3, IQR 1 – 11.5 vs 0, IQR 0-1 for non-responders and responders, respectively). 501 Thus, it may be speculated that in NMBC, clusters formation is related to the presence and 502 characteristics of primary tumor, and the neoadjuvant treatment has a different effect on the primary 503 tumor and on clusters. Discussion 431 In this context, 466 simpler technologies, as those based on size-exclusion would represent a more affordable approach, 467 easily transferable to clinical studies that might help in elucidating the role of CTC-clusters in 468 different clinical contexts. 469 Indeed, here we applied an easy-to-use filtration-based approach to investigate the relevance of 470 CTC-clusters in 37 NMBC and in 23 MBC patients. The ScreenCell® technology was chosen since 471 its validity has already been reported both for single CTCs and for clusters both when identified 472 based on cytomorphological criteria only, [37, 38] or based on marker expression [39]. 473 Overall we report that, in baseline samples collected at the beginning of NAC, the detection of at 474 least 1 CTC-cluster occurred at least 3-times more frequently in women with early breast cancer than 475 in women beginning first line treatment for MBC (a result that we also observed in our previous pilot 476 study, which was comparing ScreenCell® with AdnaTest technology [30]). Although, due to the 477 small case series, we have not done a formal analysis to exclude a bias due to different distribution of 478 molecular subtypes between the two groups, molecular subtype linked effects would have impacted 479 the data in opposite direction than observed. Thus our findings support the concept that dissemination 480 of CTC-clusters is an early event in NMBC patients, rather than an event occurring during metastatic 481 progression, as might have been expected by the high metastatic potential of clusters [4]. Since 482 dissemination is proven to occur early in breast cancer [8, 9], and indeed single CTCs hold 483 prognostic value also in NMBC women [11-13], the more frequent presence of CTC-clusters and the 484 higher number of clusters seen in early rather than later steps of the disease is intriguing. 485 22 Nevertheless, many questions on clinical and biological aspects still remain to be answered. We 486 observed that molecular subtypes affect the prevalence of CTC-clusters. In particular CTC-clusters 487 were found to be significantly more frequent in women bearing HER2-negative tumors, a result that 488 may appear as contra-intuitive since HER2-positive tumors are more aggressive and are frequently 489 associated with stemness markers [40]. Discussion 431 Moreover, despite this study is not properly powered to detect differences in 504 disease-free survival and no association was observed between relapse and CTC-clusters at baseline, 505 it is intriguing to think of potential applications of cluster enumeration after surgery as a completion 506 of pathological staging to assess the overall combined response to systemic and locoregional 507 treatments. 508 Notably, a discrepancy between cluster dynamics and imaging was observed. As consistently shown 509 by the index cases, clusters generally increased during NAC notwithstanding the concomitant 510 23 radiological and metabolic response. On the other hand, patients that did not show response to NAC 511 had a significantly higher number of clusters after surgery. This suggests a more nuanced role of 512 clusters in NMBC with respect to that of epithelial clusters in the metastatic setting. 513 radiological and metabolic response. On the other hand, patients that did not show response to NAC 511 had a significantly higher number of clusters after surgery. This suggests a more nuanced role of 512 clusters in NMBC with respect to that of epithelial clusters in the metastatic setting. 513 A crucial question raised by our results deals with the phenotype and the actual composition of 514 clusters. Having used a size-based approach for cluster enrichments and morphological criteria for 515 the detection, we were confident about the malignancy of the clusters, but we lost the information 516 regarding their epithelial/non-epithelial phenotype. However, since in the numerous studies run with 517 the CellSearch® in women with early disease, massive presence of clusters has not been reported, we 518 speculate that clusters detected in the current study are not frankly epithelial, but rather with a 519 mesenchymal or with a mixed phenotype. 520 A crucial question raised by our results deals with the phenotype and the actual composition of 514 clusters. Having used a size-based approach for cluster enrichments and morphological criteria for 515 the detection, we were confident about the malignancy of the clusters, but we lost the information 516 regarding their epithelial/non-epithelial phenotype. However, since in the numerous studies run with 517 the CellSearch® in women with early disease, massive presence of clusters has not been reported, we 518 speculate that clusters detected in the current study are not frankly epithelial, but rather with a 519 mesenchymal or with a mixed phenotype. Conclusions 535 This study represents a small snapshot on CTC-cluster detection methods and on the prevalence of 536 clusters in BC patients at different disease stages. Nonetheless, it highlights the possible bias linked 537 to inadequate methods for cluster detection, a technical bias that is worth to be considered in future 538 translational studies. In addition, we report a new observation on the fact that CTC-clusters are 539 frequent in women with NMBC. This represents a provocative finding that needs to be addressed in 540 future studies on larger series of cancer patients, homogeneous with respect to molecular subtype. 541 Finally, the observation that CTC-clusters do not disappear during neoadjuvant treatment, foster the 542 Finally, the observation that CTC-clusters do not disappear during neoadjuvant treatment, foster the 542 importance of developing treatments specifically aimed at interfering with them. 543 Finally, the observation that CTC-clusters do not disappear during neoadjuvant treatment, foster the 542 importance of developing treatments specifically aimed at interfering with them. 543 Discussion 431 520 Regarding the cluster composition, the role of inflammatory cells remains to be addressed. Indeed, 521 cooperation and crosstalk with other blood cells play a relevant role in increasing the metastasis- 522 promoting efficiency of cluster [7, 42-45]. However, in the current study, we did not find an 523 association between TILs evaluated on the primary tumor and CTC-clusters, thus the possible 524 interaction between inflammatory cells and CTC-clusters warrants further studies. 525 Finally, the observation that clusters do not disappear with the neoadjuvant treatment (and thus 526 possibly also persist after adjuvant treatment), support the need to develop treatment strategies 527 specifically designed at interfering with clusters [6, 46]. Such strategies would be promising 528 especially if CTC-clusters isolated in NMBC patients would prove to hold metastatic potential, a still 529 unanswered question worth to be addressed in the future. 530 We are aware of the study limits due to the small size and heterogeneity of the case series, although 531 its strength may be linked to the fact that these represent real-world patients, prospectively collected 532 within the daily clinical practice. 533 534 24 24 Ethics statement 557 The studies involving human participants were reviewed and approved by Institutional Review Board 558 and Ethics Committee of Fondazione IRCCS Istituto Nazionale dei Tumori on February 19, 2013, 559 and by the Institutional Review Board at the Robert H. Lurie Comprehensive Cancer Center of 560 Northwestern University (NUDB16Z01). The patients/participants provided their written informed 561 consent to participate in this study. 562 Consent for publication 563 Not Applicable. 564 Not Applicable. 564 Data Availability Statement 565 The datasets used and/or analysed during the current study are available from the corresponding 566 author on reasonable request. 567 The datasets used and/or analysed during the current study are available from the corresponding 566 author on reasonable request. 567 Funding 570 Financial support was received from Associazione Italiana per la Ricerca sul Cancro, AIRC (IG 571 16900 - M.G. Daidone; IG 20774– S. Di Cosimo); European Commission under the 7th Framework 572 Programme, grant agreement N. 260791 Eurocan Platform. C.R. is the recipient of an AIRC 573 fellowship (N. 23916). 574 Competing interests 568 All authors declare no competing interests. 569 List of abbreviations 544 BC: breast cancer; CK: cytokeratin; CTC: circulating tumor cell; DT: during treatment; 545 BC: breast cancer; CK: cytokeratin; CTC: circulating tumor cell; DT: during treatment; 545 BC: breast cancer; CK: cytokeratin; CTC: circulating tumor cell; DT: during treatment; 545 EMT: epithelial-to-mesenchymal transition; EOT: end of treatment; HSA: human serum albumin; 546 IQR: interquartile range; IS: isolation support; MBC: metastatic breast cancer; MRI: magnetic 547 resonance imaging; NAC: neoadjuvant chemotherapy; NMBC: non-metastatic breast cancer; 548 OS: overall survival; pCR: pathological complete response; PBS: phosphate-buffered saline; 549 PD: progressive disease; PET: positron emission tomography; PFS: progression free survival; 550 PR: partial response; SD: stable disease; SUV: standardized uptake value; TIL: tumor-infiltrating 551 lymphocytes. 552 555 25 Authors’ Contributions 575 Conception and design of the study: V.C.; C.R.; S.D.C.; M.G.D. 576 26 Acquisition of data: C.R.; S.D.C.; R.M.; A.M.; M.V.; A.V.; P.D.; Y.Z.; C.D.; G.S.; S.F.; G.P. 577 Analysis and/or interpretation of data: C.R.; S.D.C.; L.G.; M.C.; V.C.; M.G.D. 578 Drafting the manuscript: C.R.; V.C.; L.G.; S.D.C. 579 Revising the manuscript critically: R.M.; A.V.; P.D.; Y.Z.; M.V.; A.M.; C.D.; G.S.; S.F.; G.P.; M.C.; 580 M.G.D. 581 All authors approved the final version of the manuscript. 582 Acknowledgments 583 We are really grateful to all the patients who donated their blood for this study. 584 We thank Dr. Patrizia Miodini for skillful technical assistance in sample processing and Mrs. Cinzia 585 De Marco for preparing the mammospheres. 586 Fondazione Banca del Monte di Lombardia supported C.R. for a six-months visit to the Northwestern 587 University of Chicago to carry the technological comparison experiments. 588 We thank the Lynn Sage Cancer Research Foundation and the The Foundation Blanceflor 589 Boncompagni Ludovisi, née Bildt that supported this study. 590 Janine Wechsler (ScreenCell, Sarcelles, France) counted the CTC-clusters present on ScreenCell 591 filters, without knowledge of the clinical data. 592 Daniel Adams provided technical support for cluster detection on CellSieve filters. 593 We would like to dedicate this article to the memory of Veronica Graziani and to thank Monica 594 Remiddi and Paolo Graziani for their activity in supporting cancer research. 595 596 597 Acquisition of data: C.R.; S.D.C.; R.M.; A.M.; M.V.; A.V.; P.D.; Y.Z.; C.D.; G.S.; S.F.; G.P. 577 Analysis and/or interpretation of data: C.R.; S.D.C.; L.G.; M.C.; V.C.; M.G.D. 578 Drafting the manuscript: C.R.; V.C.; L.G.; S.D.C. 579 Revising the manuscript critically: R.M.; A.V.; P.D.; Y.Z.; M.V.; A.M.; C.D.; G.S.; S.F.; G.P.; M.C.; 580 M.G.D. 581 All authors approved the final version of the manuscript. 582 Analysis and/or interpretation of data: C.R.; S.D.C.; L.G.; M.C.; V.C.; M.G.D. 578 Drafting the manuscript: C.R.; V.C.; L.G.; S.D.C. 579 Revising the manuscript critically: R.M.; A.V.; P.D.; Y.Z.; M.V.; A.M.; C.D.; G.S.; S.F.; G.P.; M.C.; 580 M.G.D. 581 All authors approved the final version of the manuscript. 582 Acknowledgments 583 We are really grateful to all the patients who donated their blood for this study. 584 We thank Dr Patrizia Miodini for skillful technical assistance in sample processing and Mrs Cinzia 585 Analysis and/or interpretation of data: C.R.; S.D.C.; L.G.; M.C.; V.C.; M.G.D. 578 Acknowledgments 583 We are really grateful to all the patients who donated their blood for this study. 584 We thank Dr. Patrizia Miodini for skillful technical assistance in sample processing and Mrs. Cinzia 585 De Marco for preparing the mammospheres. 586 We thank Dr. Patrizia Miodini for skillful technical assistance in sample processing and Mrs. Cinzia 585 De Marco for preparing the mammospheres. 586 Fondazione Banca del Monte di Lombardia supported C.R. for a six-months visit to the Northwestern 587 University of Chicago to carry the technological comparison experiments. 588 Fondazione Banca del Monte di Lombardia supported C.R. for a six-months visit to the Northwestern 587 University of Chicago to carry the technological comparison experiments. 588 We thank the Lynn Sage Cancer Research Foundation and the The Foundation Blanceflor 589 Boncompagni Ludovisi, née Bildt that supported this study. 590 We thank the Lynn Sage Cancer Research Foundation and the The Foundation Blanceflor 589 Boncompagni Ludovisi, née Bildt that supported this study. 590 Janine Wechsler (ScreenCell, Sarcelles, France) counted the CTC-clusters present on ScreenCell 591 filters, without knowledge of the clinical data. 592 Janine Wechsler (ScreenCell, Sarcelles, France) counted the CTC-clusters present on ScreenCell 591 filters, without knowledge of the clinical data. 592 Daniel Adams provided technical support for cluster detection on CellSieve filters. 593 Daniel Adams provided technical support for cluster detection on CellSieve filters. 593 We would like to dedicate this article to the memory of Veronica Graziani and to thank Monica 594 Remiddi and Paolo Graziani for their activity in supporting cancer research. 595 We would like to dedicate this article to the memory of Veronica Graziani and to thank Monica 594 Remiddi and Paolo Graziani for their activity in supporting cancer research. 595 27 References 601 1. 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Oncotarget. 7(17):24677-87. doi: 708 10.18632/oncotarget.8250 709 35. Sarioglu AF, Aceto N, Kojic N, Donaldson MC, Zeinali M, Hamza B, et al. (2017) A 710 microfluidic device for label-free, physical capture of circulating tumor cell clusters. Nat Methods. 711 12(7):685-91. doi: 10.1038/nmeth.3404. 712 30 36. Piñeiro R, Martínez-Pena I, López-López R. (2020) Relevance of CTC Clusters in Breast 713 Cancer Metastasis. Adv Exp Med Biol. 1220:93-115. doi: 10.1007/978-3-030-35805-1_7. 714 36. Piñeiro R, Martínez-Pena I, López-López R. (2020) Relevance of CTC Clusters in Breast 713 37. Mascalchi M, Maddau C, Sali L, Bertelli E, Salvianti F, Zuccherelli S, et al. (2017) 715 Circulating tumor cells and microemboli can differentiate malignant and benign pulmonary 716 lesions. Cancer 8(12):2223-2230. doi: 10.7150/jca.18418. eCollection 717 38. O'Toole SA, Spillane C, Huang Y, Fitzgerald MC, Ffrench B, Mohamed B, et al. (2020) 718 Circulating tumour cell enumeration does not correlate with Miller-Payne grade in a cohort of breast 719 cancer patients undergoing neoadjuvant chemotherapy. Breast Cancer Res Treat. 181(3):571-580. 46. Heeke S, Mograbi B, Alix-Panabières C, Hofman P. (2019) Never Travel Alone: The 744 Crosstalk of Circulating Tumor Cells and the Blood Microenvironment. Cells. 8(7):714. doi: 745 10.3390/cells8070714. 746 46. Heeke S, Mograbi B, Alix-Panabières C, Hofman P. (2019) Never Travel Alone: The 744 Crosstalk of Circulating Tumor Cells and the Blood Microenvironment. Cells. 8(7):714. doi: 745 46. Heeke S, Mograbi B, Alix-Panabières C, Hofman P. (2019) Never Travel Alone: The 744 Crosstalk of Circulating Tumor Cells and the Blood Microenvironment. Cells. 8(7):714. doi: 745 References 601 720 doi: 10.1007/s10549-020-05658-7. 721 39. Mu Z, Benali-Furet N, Uzan G, Znaty A Ye Z, Paolillo C, et al. (2016) Detection and 722 Characterization of Circulating Tumor Associated Cells in Metastatic Breast Cancer. Int J Mol Sci. 723 17(10):1665. doi: 10.3390/ijms17101665. 724 40. Korkaya H, Kim GI, Davis A, Malik F, Henry NL, Ithimakin S, et al. (2012) Activation of 725 an IL6 inflammatory loop mediates trastuzumab resistance in HER2+ breast cancer by expanding the 726 cancer stem cell population. Mol Cell. 47(4):570-84. doi: 10.1016/j.molcel.2012.06.014. 727 41. Gawrzak S, Rinaldi L, Gregorio S, Arenas EJ, Salvador F, Urosevic J, et al. (2018) MSK1 728 regulates luminal cell differentiation and metastatic dormancy in ER(+) breast cancer. Nat Cell Biol. 729 20(2):211-221. doi: 10.1038/s41556-017-0021-z. 730 42. Labelle M, Begum S, Hynes RO. (2011) Direct signaling between platelets and cancer 731 cells induces an epithelial-mesenchymal-like transition and promotes metastasis. Cancer Cell. 732 20(5):576-90. doi: 10.1016/j.ccr.2011.09.009. 733 43. Hamilton G, Rath B. (2017) Circulating tumor cell interactions with macrophages: 734 implications for biology and treatment. Transl Lung Cancer Res. 6(4):418-430. doi: 735 10.21037/tlcr.2017.07.04. 736 44. Arnoletti JP, Fanaian N, Reza J, Sause R, Almodovar AJ, Srivastava M, et al. (2018) 737 Pancreatic and bile duct cancer circulating tumor cells (CTC) form immune-resistant multi-cell type 738 clusters in the portal venous circulation. Cancer Biol Ther. 19(10):887-897. doi: 739 10.1080/15384047.2018.1480292. 740 45. Sprouse ML, Welte T, Boral D, Liu HN, Yin W, Vishnoi M, et al. (2019) PMN-MDSCs 741 Enhance CTC Metastatic Properties through Reciprocal Interactions via ROS/Notch/Nodal Signaling. 742 Int J Mol Sci. 20(8):1916. doi: 10.3390/ijms20081916. 743 46. Heeke S, Mograbi B, Alix-Panabières C, Hofman P. (2019) Never Travel Alone: The 744 Crosstalk of Circulating Tumor Cells and the Blood Microenvironment. Cells. 8(7):714. doi: 745 10.3390/cells8070714. 746 10.3390/cells8070714. 746 749 31 Figure 1. Comparison of CellSearch and CellSieve for CTC-cluster detection in clinical 751 samples. 752 Figure 1. Comparison of CellSearch and CellSieve for CTC-cluster detection in clinical 751 samples. 752 (A) Nineteen blood samples collected from patients with MBC were processed in parallel with 753 CellSearch and CellSieve for the detection of CTC-clusters. (B) Representative image of a CKpos 754 CTC-cluster detected by CellSearch (green = CK; pink = DAPI; 10X magnification). 755 (C-D) Representative images of a CKpos (C) and a CKneg (D) cluster detected by CellSieve 756 (green = CK; blue = DAPI; yellow = CD45; the white arrows indicate 2 CD45neg/CKneg cells inside 757 the cluster). (E) Doughnut plot showing the percentages of samples containing CKpos CTC-clusters 758 (blue) analyzed by CellSearch (outer circle, 53 %) and CellSieve (inner circle, 79 %). Positivity 759 threshold was set at 1 CTC-cluster/7.5 mL of blood. The percentage of CellSieve samples containing 760 only CKneg clusters are shown in orange (5 %). (F) Spaghetti plot showing the numbers of 761 CTC-clusters detected in each sample analyzed by CellSearch and CellSieve. For CellSieve samples, 762 both the counts of CKpos CTC-clusters only and of CKpos plus CKneg clusters (CellSieve total) are 763 reported (colors are arbitrary assigned for increasing readability of the graph only). 764 Detection of CTC-clusters in patients with early and metastatic breast cancer. (A) Representative images of CTC-clusters enriched by filtration using ScreenCell filters. The list of 766 cytomorphological criteria used for the identification of CTC-clusters is reported in the inset. 767 (B – D) Boxplots reporting the number of CTC-clusters detected in baseline samples collected from 768 NMBC vs. MBC patients (B); and in baseline samples collected from NMBC patients, according to 769 the patients’ nodal status (C) and to the disease subtype (D). 770 771 772 32 32 Figure 3. CTC-cluster evaluation during neoadjuvant therapy in early breast cancer patients. 773 (A) Violin plot showing the number of CTC-clusters detected in samples longitudinally collected 774 from 37 NMBC patients. CTC-clusters were evaluated before starting neoadjuvant treatment 775 (Baseline), during (DT), at the end of therapy (EOT), and after surgery (Surgery). The colors indicate 776 the BC subtype (blue = HER2-positive; purple = luminal-like; red = triple-negative) while the gray 777 shadow indicates the density of samples for the corresponding CTC-cluster number. The detailed 778 description of 2 index cases is reported in panels B and C. TNBC = triple-negative breast cancer; 779 AT = Antracyclines, Taxanes; CMF = Cyclophosphamide, Methotrexate, Fluorouracil; 780 pCR = pathological complete response. 781 Additional file 1 (.DOCX): Table reporting the clinico-pathological characteristics of the patients’ 784 cohort for technological comparison. 785 Additional file 2 (.TIFF): Figure showing CTC-clusters expressing CD31. 786 Additional file 2 (.TIFF): Figure showing CTC-clusters expressing CD31. 786 Additional file 3 (.DOCX): Table reporting CTC-cluster count in samples processed in parallel with 787 CellSearch and CellSieve filters. 788 Additional file 3 (.DOCX): Table reporting CTC-cluster count in samples processed in parallel with 787 CellSearch and CellSieve filters. 788 Additional file 4 (.TIFF): Figure showing the association of CTC-clusters with clinico-pathological 789 characteristics in patients with NMBC. 790 Additional file 4 (.TIFF): Figure showing the association of CTC-clusters with clinico-pathological 789 characteristics in patients with NMBC. 790 Additional file 5 (.TIFF): Figure showing the association of CTC-clusters with outcome in NMBC 791 patients. 792 Additional file 5 (.TIFF): Figure showing the association of CTC-clusters with outcome in NMBC 791 patients. 792 793 33 Figures Figures Figure 1 Comparison of CellSearch and CellSieve for CTC-cluster detection in clinical s l ll t d f ti t ith MBC d i ll l ith C Figures Figure 1 Comparison of CellSearch and CellSieve for CTC-cluster detection in clinical samples. (A) Nineteen blood samples collected from patients with MBC were processed in parallel with CellSearch and CellSieve for the detection of CTC-clusters. (B) Representative image of a CKpos CTC-cluster detected by CellSearch Figure 1 Figure 1 Comparison of CellSearch and CellSieve for CTC-cluster detection in clinical samples. (A) Nineteen blood samples collected from patients with MBC were processed in parallel with CellSearch and CellSieve for the detection of CTC-clusters. (B) Representative image of a CKpos CTC-cluster detected by CellSearch (green = CK; pink = DAPI; 10X magni¦cation). (C-D) Representative images of a CKpos (C) and a CKneg (D) cluster detected by CellSieve (green = CK; blue = DAPI; yellow = CD45; the white arrows indicate 2 CD45neg/CKneg cells inside the cluster). (E) Doughnut plot showing the percentages of samples containing CKpos CTC-clusters (blue) analyzed by CellSearch (outer circle, 53 %) and CellSieve (inner circle, 79 %). Positivity threshold was set at 1 CTC-cluster/7.5 mL of blood. The percentage of CellSieve samples containing only CKneg clusters are shown in orange (5 %). (F) Spaghetti plot showing the numbers of CTC-clusters detected in each sample analyzed by CellSearch and CellSieve. For CellSieve samples, both the counts of CKpos CTC-clusters only and of CKpos plus CKneg clusters (CellSieve total) are reported (colors are arbitrary assigned for increasing readability of the graph only). (green = CK; pink = DAPI; 10X magni¦cation). (C-D) Representative images of a CKpos (C) and a CKneg (D) cluster detected by CellSieve (green = CK; blue = DAPI; yellow = CD45; the white arrows indicate 2 CD45neg/CKneg cells inside the cluster). (E) Doughnut plot showing the percentages of samples containing CKpos CTC-clusters (blue) analyzed by CellSearch (outer circle, 53 %) and CellSieve (inner circle, 79 %). Positivity threshold was set at 1 CTC-cluster/7.5 mL of blood. The percentage of CellSieve samples containing only CKneg clusters are shown in orange (5 %). (F) Spaghetti plot showing the numbers of CTC-clusters detected in each sample analyzed by CellSearch and CellSieve. For CellSieve samples, both the counts of CKpos CTC-clusters only and of CKpos plus CKneg clusters (CellSieve total) are reported (colors are arbitrary assigned for increasing readability of the graph only). (green = CK; pink = DAPI; 10X magni¦cation). (C-D) Representative images of a CKpos (C) and a CKneg (D) cluster detected by CellSieve (green = CK; blue = DAPI; yellow = CD45; the white arrows indicate 2 CD45neg/CKneg cells inside the cluster). (E) Doughnut plot showing the percentages of samples containing CKpos CTC-clusters (blue) analyzed by CellSearch (outer circle, 53 %) and CellSieve (inner circle, 79 %). Positivity threshold was set at 1 CTC-cluster/7.5 mL of blood. Figure 1 The percentage of CellSieve samples containing only CKneg clusters are shown in orange (5 %). (F) Spaghetti plot showing the numbers of CTC-clusters detected in each sample analyzed by CellSearch and CellSieve. For CellSieve samples, both the counts of CKpos CTC-clusters only and of CKpos plus CKneg clusters (CellSieve total) are reported (colors are arbitrary assigned for increasing readability of the graph only). Figure 2 Detection of CTC-clusters in patients with early and metastatic breast cancer. (A) Representative im Figure 2 Detection of CTC-clusters in patients with early and metastatic breast cancer. (A) Representative images of CTC-clusters enriched by ¦ltration using ScreenCell ¦lters. The list of cytomorphological criteria used for the identi¦cation of CTC-clusters is reported in the inset. (B – D) Boxplots reporting the number of CTC-clusters detected in baseline samples collected from NMBC vs. MBC patients (B); and in baseline Figure 2 Detection of CTC-clusters in patients with early and metastatic breast cancer. (A) Representative images of CTC-clusters enriched by ¦ltration using ScreenCell ¦lters. The list of cytomorphological criteria used for the identi¦cation of CTC-clusters is reported in the inset. (B – D) Boxplots reporting the number of CTC-clusters detected in baseline samples collected from NMBC vs. MBC patients (B); and in baseline samples collected from NMBC patients, according to the patients’ nodal status (C) and to the disease subtype (D). samples collected from NMBC patients, according to the patients’ nodal status (C) and to the disease subtype (D). samples collected from NMBC patients, according to the patients’ nodal sta subtype (D). Figure 3 CTC-cluster evaluation during neoadjuvant therapy in early breast cancer pa the number of CTC-clusters detected in samples longitudinally collected fro f ( ) Figure 3 CTC-cluster evaluation during neoadjuvant therapy in early breast cancer patients. (A) Violin plot showing the number of CTC-clusters detected in samples longitudinally collected from 37 NMBC patients. CTC- clusters were evaluated before starting neoadjuvant treatment (Baseline), during (DT), at the end of Figure 3 Figure 3 Figure 3 Figure 3 CTC-cluster evaluation during neoadjuvant therapy in early breast cancer patients. (A) Violin plot showing the number of CTC-clusters detected in samples longitudinally collected from 37 NMBC patients. CTC- clusters were evaluated before starting neoadjuvant treatment (Baseline), during (DT), at the end of therapy (EOT), and after surgery (Surgery). The colors indicate the BC subtype (blue = HER2-positive; purple = luminal-like; red = triple-negative) while the gray shadow indicates the density of samples for the corresponding CTC-cluster number. The detailed description of 2 index cases is reported in panels B and C. TNBC = triple-negative breast cancer; AT = Antracyclines, Taxanes; CMF = Cyclophosphamide, Methotrexate, Fluorouracil; pCR = pathological complete response. Supplementary Files This is a list of supplementary ¦les associated with this preprint. Click to download. Additional¦le1.docx Additional¦le2.tiff Additional¦le3.docx Additional¦le4.tiff Additional¦le5.tiff
https://openalex.org/W2098484026	https://zenodo.org/records/2298276/files/article.pdf	English	null	EXTENSION OF THE OLD PIER AT HARTLEPOOL.	Minutes of proceedings	1,912	public-domain	1,956	Vide “ History of Hartlepool,” Sir Cuthbert Sharp, Hartlepool, 1816. 413 413 Papers.] HOWKINS ON EXTENSION OF OLD PIER, HARTLEPOOL. By JOHN DRTSDALE HOWKINS, M A . , M. Inst. C.E. (Abridged.) AMONG the extensive improvements carried out at the Port of Hartlepool during the last few years by the North Eastern Railway Company and the Hartlepool Port and Harbour Commission, the Author proposes to select for description in this Paper one particular work, namely, the extension of the Old Pier ; first, because it is one of the oldest of British sea-works in existence at the present day, and secondly, because the method of building the new pier-head by means of interlocking concrete blocks is somewhat unusual. g The title of Old Pier is no misnomer, as there is evidence that it wasoriginally built in 1473 in response to an appeal for funds by Bishop Booth of Durham.l Its existence was somewhat precarious until an -4ct of Parliament in 1851 placed it under the control of the Hartlepool Port and Harbour Commission. The increase in the depth of the channel at Hartlepool from 10 feet in 1902 to 20 feet at the present day has undoubtedly permitted much heavier seas to strike the Old Pier than formerly, and the end of this work became so shaken that in the Spring of 1909 lit was decided that it could no longer stand repair, and that the building of a new hea.d was imperative. I n construction it resembles many other old sea-works, with drystone wall of rough limestone, built to a batter of about 1 in 4 and backed by rubble hearting. It was decided to construct the new head outside the old one. The new head (Figs. 1 ) is built of interlocking concrete blocks, weighing approximately 12 cwt. each, about the maximum weight which can be easily handled without steam-power. These blocks are rectangular in plan where the wall is straight, but have radiating Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. HOWKINS OS THE EXTENSION OF 414 [Selected I,’ Y T m Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. 41 5 THE OLD PIER AT HARTLEPOOL. Papers.] joints where it is curved. The radius of the face is 36 feet. The work is founded on the clay, which was met with a t a depth varying between 2 and 4 feet below low-water mark. Considerable difficulty was experienced in making the excavation at the lower depths, as this work could only be carried out during extraordinary spring tides; and the northerly winds which prevailed in 1909 and 1910 frequently held up the ebbs. The work, which was carried out entirely by the Commissioners’ own workmen, was started in April, 1909, by making a number of concrete protection blocks, each of 12 tons weight, on the top of the old structure. As soon as these blocks were sufficiently hard, they were moved forward on railway rails by means of screw-jacks and launched into the sea at high tide, in order that their fall might be broken as much as possible. It was found that, with one or two exceptions, they took up their proper position just beyond the line of the foundation. This was fortu- nate, as the moving with screw-jacks of blocks of this weight half embedded in clay in the limited time at low water was a very arduous work. BZoe?cyurd.-In the meantime, the small blockyard was being constructed. It was situated on the beach to the east of the shore end of the Old Pier, and connected by a line of 20-inch gauge railway with the head. The blocks were made in batteries of sixteen each in the case of headers, and of ten each in the case of stretchers, in order both to economise woodwork as much as possible, and also to produce blocks of a more uniform size than if separate moulds had been used. Below the blockyard on the beach was an ample supply of suitable shingle and sand in about the proper proportions for concrete. This was piled into a stockheap beside the blockyard, and was delivered to the hand-mixing machine from small side-tip wagons. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. The first batches of blocks were made of concrete in the proportion of 6 of gravel and sand to 1 of cement, as these blocks, being required for the lowest courses, would have less time to mature than those made subsequently, and would naturally have to resist a greater stress. The proportions in other blocks were :-face-blocks, 7 to 1, back-blocks, 85 to 1. Excavation.-Excavation for the foundations could only be carried out during low water of spring tides, and double tides were worked whenever possible. As only a short length could thus be got out at a time, concrete was at once deposited. Bays were formed of con- crete in hop bags, and mass-concrete was deposited in the tranquil water within these bays, To counteract any tendency of the foundation to slide forward on tbe day bed, a vertical step was Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. 416 HOWKINS ON THE EXTENSION OF [Selected formed half-way across the section so that the back of the foundation was slightly deeper than the front,. formed half-way across the section so that the back of the foundation was slightly deeper than the front,. Deterioration of Concrete.-A firm foundation on clay was met with, except a t one point on the inside of the pier, where the black colour and the smell of the mud between the large boulders suggested the presence of a sulphur spring. It ihad been previously ascertained that, when repairs to the old head at this point had been carried out, the concrete used rapidly deteriorated, this being the only instance which has come under the observation of the Author of concrete made from sound materials suffering from other than accidental causes in a sea-work. By the courtesy of Messrs. Otto Trechmann, the local cement manufacturers, two analyses were made :-the first, of some concrete which had deteriorated when brought into conjunction with this mud, and the second of the mud itself. These analyses indicated that the presence of the black mud itself did not cause the deterioration of the concrete, but that this was brought about by gypsiferous water from a bed of anhydrite and gypsum which occurs in the magnesian limestone below the harbour, and abuts against that rock beneath the boulder-clay near the end of the Old Pier. It is interesting to note, first, that the old inhabit- ants of Hartlepool used to obtain a medicinal water near the Old Pier, and secondly, that Messrs. Otto Trechmann, when boring at their works about a mile distant, met with this same gypsiferous water after piercing 265 feet of anhydrite. It was of course considered undesirable to use concrete for this part of the foundation, and large blocks of stone from the demolition of the old Victoria Dock Entrance were substituted. Blocks.-The concrete blocks were set by a hand-derrick crane on a bedding of 4 parts of fine sand to 1 of ordinary Portland Cement, and the vertical joints were kept as close as possible, and pointed with quick-setting cement. When the latter had set, grout was poured into the lewis-holes, and over the top of the blocks so as to fill all openings. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. It had been intended to pick out the quick-setting cement from the joints, and point up afterwards with mortar made with ordinary cement if any deterioration should have taken place, but the necessity for this has not arisen so far. The total number of blocks built into the wall was approximately 2,300. Backing.-The original intention had been to use the stones from the excavation, and any rubble handy for the purpose of filling the gap between the old and new work, but the contractor for the Victoria Dock Improvement was glad to allow the stone from the old entrance to be taken away for nothing. As many coping and Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. THE OLD PIEIl AT IIARTLEPOOL. Papers.) 41 7 Papers.) other large stones, both dressed and undressed, could be obtained from this source, a much better material than rubble was thus available. All work in connection with the pier-head was corn- pleted in 1911. Removal of Lighthouse.-The addition to the Old Pier of the new head rendered the position of the lighthouse unsuitable, and, with the approval of the Elder Brethren of the Trinity House, this structure was bodily moved 30 feet seaward. Every preparation was made beforehand, and the actual moving only occupied some 30 minutes. The lighthouse is an openwork timber structure 14 feet square at the base and 40 feet in height, with a fourth- order red dioptric light. It is interesting to note that the movement was carried out so smoothly that the incandescent mantle which had been on the burner previously was employed on the same night. p y p y g New Sea- Wall.-It will have been observed from the plan of the new pier-head that the straight part was not built parallel with the east side of the Old Pier. The reason for this was that it was foreseen that in the near future the seaward side of the old wall would inevitably become undermined by the sea, and it was decided in this eventuality to build a new wall at an angle so as to include a considerable area of ground which would become valuable for the erection of buildings, etc. A protracted gale or series of gales from the E.S.E. in December, 1910, settled this question by breaching the old wall, and undermining it practically for its whole length. It was apparent from an examination of the foundation-stones that the shelling away of the flat strata of limestone rock upon which the wall is founded had been the cause of the undermining. As it was necessary to lose no time in the construction of the new wall, it was put in hand in the Spring of 1911, and is approaching com- pletion. From the plan and section (Figs. Z), it will be seen that the work is being carried out in mass-concrete instead of blockwork. Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved. The only reason for this change is that, owing to the diminishing depth, the former method of construction is more rapid and con- venient than the latter. The new wall is 375 feet in length, and is identical in section with a similar work which was carried out from the designs of the Author in 1906 for the Scarborough Harbour Commission. The Paper is accompanied by 4 drawings, from which the Figures in the text have been selected. [THE INST. C.E. VOL. CLXXXVIII.] 2 E [THE INST. C.E. VOL. CLXXXVIII.] Downloaded by [ UNIVERSITY OF EXETER] on [11/09/16]. Copyright © ICE Publishing, all rights reserved.
https://openalex.org/W2029386427	https://europepmc.org/articles/pmc3552965?pdf=render	English	null	Comprehensive Phylogenetic Analysis of Bacterial Group II Intron-Encoded ORFs Lacking the DNA Endonuclease Domain Reveals New Varieties	PloS one	2,013	cc-by	5,924	Introduction classifications overlap to some extent. This has led to proposals for a single classification based on the coevolution of the two intron components, RNA and IEP (i.e. intron class D, with a IIB-like RNA structure, is known as IIB3 in this system) [7]. According to the IEP-based phylogenetic nomenclature, bacteria contain introns from all phylogenetic classes, whereas organelles contain only introns of the CL and ML classes. Class A, C, D, E and F introns encode proteins with no En domain. It is generally thought that the En domain of group II introns was acquired by an RT/ maturase IEP [9,14], and it has recently been suggested that this domain may have been acquired only once, by a common ancestor of B, E, ML and CL introns, subsequently being lost from class E [15]. Group II introns are self-splicing RNAs that act as mobile retroelements and are thought to be the ancestors of nuclear spliceosomal introns [1–4]. Group II introns were initially identified in the mitochondrial and chloroplast genomes of lower eukaryotes and plants, but were subsequently found in bacteria and archaea [5–9]. A typical group II intron consists of a highly structured RNA that folds into a conserved three-dimensional structure consisting of six distinct double-helical domains, DI to DVI [10]. Most bacterial group II introns have a multifunctional intron-encoded protein (IEP) ORF within DIV [8]. Group II IEPs have an N-terminal reverse transcriptase (RT) domain homolo- gous to retroviral RT sequences, followed by a putative RNA- binding domain with RNA splicing or maturase activity (domain X) and, in some cases, a C-terminal DNA-binding (D) region followed by a DNA endonuclease (En) domain [11,12]. Three main classes (IIA, IIB and IIC) of group II introns have been described on the basis of conserved ribozyme structures, with additional subclasses displaying specific structural variations (i.e., IIA1, IIA2, IIB1 and IIB2) [8,13]. It is generally accepted that group II intron RNAs have coevolved with their IEPs, and phylogenetic analyses of group II intron-encoded ORFs has resulted in their classification into several groups (A, B, C, D, E, F, CL1 [chloroplast-like 1], CL2 [chloroplast-like 2] and ML [mitochondrion-like]). Nomenclatures of intron subclasses based on ribozyme structure or the IEP are widely used, but these Class F is the most recently identified bacterial ORF phyloge- netic class. Comprehensive Phylogenetic Analysis of Bacterial Group II Intron-Encoded ORFs Lacking the DNA Endonuclease Domain Reveals New Varieties Nicola´s Toro, Francisco Martı´nez-Abarca o de Ecologı´a Gene´tica, Estacio´n Experimental del Zaidı´n, Consejo Superior de Investigaciones Cientı´ficas (CSIC), Granada, Spain Abstract Competing Interests: The authors have declared that no competing interest exist. E-mail: nicolas.toro@eez.csic.es Abstract Group II introns are self-splicing RNAs that act as mobile retroelements in the organelles of plants, fungi and protists. They are also widely distributed in bacteria, and are generally assumed to be the ancestors of nuclear spliceosomal introns. Most bacterial group II introns have a multifunctional intron-encoded protein (IEP) ORF within the ribozyme domain IV (DIV). This ORF encodes an N-terminal reverse transcriptase (RT) domain, followed by a putative RNA-binding domain with RNA splicing or maturase activity and, in some cases, a C-terminal DNA-binding (D) region followed by a DNA endonuclease (En) domain. In this study, we focused on bacterial group II intron ORF phylogenetic classes containing only reverse transcriptase/maturase open reading frames, with no recognizable D/En region (classes A, C, D, E, F and unclassified introns). On the basis of phylogenetic analyses of the maturase domain and its C-terminal extension, which appears to be a signature characteristic of ORF phylogenetic class, with support from the phylogeny inferred from the RT domain, we have revised the proposed new class F, defining new intron ORF varieties. Our results increase knowledge of the lineage of group II introns encoding proteins lacking the En-domain. Citation: Toro N, Martı´nez-Abarca F (2013) Comprehensive Phylogenetic Analysis of Bacterial Group II Intron-Encoded ORFs Lacking the DNA Endonuclease Domain Reveals New Varieties. PLoS ONE 8(1): e55102. doi:10.1371/journal.pone.0055102 Editor: Luis Mene´ndez-Arias, Centro de Biologı´a Molecular Severo Ochoa (CSIC-UAM), Spain Received October 19, 2012; Accepted December 18, 2012; Published January 23, 2013 Martı´nez-Abarca. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits tion, and reproduction in any medium, provided the original author and source are credited. Copyright: 2013 Toro, Martı´nez-Abarca. This is an open-access article distributed under the terms of the Creative Commons At unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. pyright: 2013 Toro, Martı´nez-Abarca. This is an open-access article distributed under the terms of the Creative Commons Attribution restricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by research grants BIO2011-24401 from the Spanish Ministerio de Ciencia e Innovacio´n, and CSD 2009-0006 of the Consolider-Ingenio programme including ERDF (European Regional Development Funds). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. January 2013 \| Volume 8 \| Issue 1 \| e55102 Phylogenetic analyses We focused on group II intron ORF phylogenetic classes containing only reverse transcriptase (RT)/maturase open reading frames (ORFs), with no recognizable D/En region (classes A, C, D, E, F and unclassified introns), using all available sequences from the database until 07-03-2012, and the new sequences identified in this work. The phylogenetic relationships between unclassified and class F intron ORFs available from the database were first investigated by comparing phylogenetic trees estimated by maximum likelihood methods from alignments of sequences for the RT and for the X-domain plus its C-terminal extension. These alignments were generated with Geneious Pro software (Biomat- ters Ltd.) and Clustal W. Introduction It currently includes a small group of bacterial group II introns with a typical class IIB RNA secondary structure, similar in overall organization to the introns of class E [16]. However, phylogenetic analysis (RT and X domains) of class F ORFs has shown relatively low levels of bootstrap support (,70%), increasing to 73% only when the unclassified introns basal to this group are excluded [16]. Simon et al. [15] performed a large-scale phylogenetic analysis of group II introns with sequence data for both the conserved RNA structure and the intron-encoded reverse transcriptase. They found that each ORF class, including class F, was a robust clade, but that the relationship between classes remained ambiguous. Furthermore, amino-acid sequence-based January 2013 \| Volume 8 \| Issue 1 \| e55102 1 January 2013 \| Volume 8 \| Issue 1 \| e55102 PLOS ONE \| www.plosone.org Group II Introns Lacking the En Domain Figure 1. ORF phylogeny based on alignments of the amino-acid sequences of unclassified and class F intron-encoded ORFs. (A) Phylogenetic tree based on the X-domain and its C-terminal extension. (B) Phylogenetic tree based on the RT-domain. MLE consensus trees based on MSAs generated with Clustal W, using the WAG substitution model. Trees are rooted at their midpoint. Bootstrap results ($75%) are given at each node (PhyML) and the rate of amino-acid substitutions per site is shown at the bottom of each tree. Introns assigned to class F are indicated and highlighted in bold typeface. doi:10.1371/journal.pone.0055102.g001 Figure 1. ORF phylogeny based on alignments of the amino-acid sequences of unclassified and class F intron-encoded ORFs. (A) Phylogenetic tree based on the X-domain and its C-terminal extension. (B) Phylogenetic tree based on the RT-domain. MLE consensus trees based on MSAs generated with Clustal W, using the WAG substitution model. Trees are rooted at their midpoint. Bootstrap results ($75%) are given at each node (PhyML) and the rate of amino-acid substitutions per site is shown at the bottom of each tree. Introns assigned to class F are indicated and highlighted in bold typeface. doi:10.1371/journal.pone.0055102.g001 phylogenies were found to be sensitive to taxon sampling, with similar sets of introns often producing different tree topologies with different levels of support for the conflicting nodes. Introduction In the most recent compilation of group II introns by Zimmerlys group [17], which was based on 397 bacterial ORF-encoding introns (sequences .95% identical are given the same name and listed only once within a given species) from the database for bacterial group II introns (http://webapps2.ucalgary.ca/,groupii/index. html#), class F contained only seven introns, whereas 10 unclassified introns also lacked a recognizable D/En region. phylogenies were found to be sensitive to taxon sampling, with similar sets of introns often producing different tree topologies with different levels of support for the conflicting nodes. In the most recent compilation of group II introns by Zimmerlys group [17], which was based on 397 bacterial ORF-encoding introns (sequences .95% identical are given the same name and listed only once within a given species) from the database for bacterial group II introns (http://webapps2.ucalgary.ca/,groupii/index. html#), class F contained only seven introns, whereas 10 unclassified introns also lacked a recognizable D/En region. (Table S1). We also searched for new types of RT/maturase ORFs lacking the endonuclease domain, using the amino-acid consensus sequence obtained from the MSA of domain X and the C-terminal extension for reported new class F and unclassified introns downloaded from the database for bacterial group II introns. The search was carried out as for the identification of class D ORFs, but with BlastP only. The additional sequences identified in the first search were then included in the alignment to obtain a refined consensus sequence, which was used to identify more distant relatives. All the previously reported class F and unclassified introns were identified among the Blast hits, together with 27 new, related ORF sequences. The final dataset for phylogenetic analysis included 50 ORF sequences that were #75% identical (Table S2). All these analyses were carried out with Geneious Pro software (Biomatters Ltd.). We previously showed that the C-terminal portion of class D RT/maturase proteins constitutes a distinctive, characteristic signature of the ORF class, which has been conserved throughout evolution. It constitutes a group-specific functionally important protein region contributing to both maturase function and intron mobility [18]. Here, on the basis of consensus maturase and C- terminal amino-acid sequences, with additional support from the phylogeny inferred from the larger RT-domain, we extend current knowledge of intron ORF subclasses with no recognizable D/En region. Our results call into question the proposed new class F, but reveal the existence of additional group II intron ORF varieties. January 2013 \| Volume 8 \| Issue 1 \| e55102 January 2013 \| Volume 8 \| Issue 1 \| e55102 January 2013 \| Volume 8 \| Issue 1 \| e55102 Group II Introns Lacking the En Domain WAG and LG) delivered trees with the same topology. The GUIDANCE method [21] was used to determine alignment confidence scores and to remove aligned positions considered unreliable according to a given cutoff. GUIDANCE is freely available for use from http://guidance.tau.ac.il. The species delimitation plugin for Genious [22] was used to calculate the inter-species divergence and Rosenbergs reciprocal monophyly P(AB), a test for taxonomic distinctiveness, according to the null hypothesis that monophyly is a chance outcome under a model of random coalescence in a single group [23]. Other types of analysis were performed to investigate the influence of the various phylogenetic estimation methods on our results. We carried out Bayesian analysis with the parallel version of MrBayes 3.1 [24] implemented in Geneious Pro (Biomatters Ltd.). Two independent runs of four chains were completed for 1,100,000 Metropolis-coupled Markov chain Monte Carlo (MCMCMC) generations, using the default priors for model parameters, the WAG model as the rate matrix (fixed) and the gamma model for between-site rate variation. Trees were sampled every 200 generations, and 110,000 samples were discarded as the ‘‘burn-in’’, to produce a 50% majority-rule consensus tree. Figure 3. RT domain-based phylogeny of group II intron ORFs. To the MSA alignment used for Fig. 2B, representatives of classes B: En.fmI3, B.a.I2, En.fm.I1, E.f.I3, B.me.I1, C.d.I1, G.k.I1, and B.c.I5; CL1A: A.v.I1, P.p.I2, E.c.I5, Th.e.I1, B.t.I1, Sh.sp.I1, Ms.b.I1; CL1B: B.me.I2, and B.a.I1; CL2A: Tr.e.I2, G.v.I1, Ns.p.I4, Cs.p.I1, and C.w.I1; CL2B: C.w.I6, Ns.p.I2, and Ns.p.I1; and ML: Ba.fr.I1, B.t.I2, N.a.I2, and L.l.I1 used by Simon et al. [15] were added (RT 0– 7). The corresponding accession numbers can be obtained from the bacterial group II intron database. A consensus unrooted tree is shown and major ORF classes and the new groups are labeled. Bootstrap results ($75%) are given for each node (PhyML). The arrow indicates the common node of the lineage of group II introns with an En domain in their IEPs. The circle indicates the possible node grouping together class F introns. doi:10.1371/journal.pone.0055102.g003 Figure 3. RT domain-based phylogeny of group II intron ORFs. To the MSA alignment used for Fig. 2B, representatives of classes B: En.fmI3, B.a.I2, En.fm.I1, E.f.I3, B.me.I1, C.d.I1, G.k.I1, and B.c.I5; CL1A: A.v.I1, P.p.I2, E.c.I5, Th.e.I1, B.t.I1, Sh.sp.I1, Ms.b.I1; CL1B: B.me.I2, and B.a.I1; CL2A: Tr.e.I2, G.v.I1, Ns.p.I4, Cs.p.I1, and C.w.I1; CL2B: C.w.I6, Ns.p.I2, and Ns.p.I1; and ML: Ba.fr.I1, B.t.I2, N.a.I2, and L.l.I1 used by Simon et al. Materials and Methods Search for additional class D and new types of RT/ maturase ORFs lacking the endonuclease domain We searched non redundant (GenBank, RefSeq, EMBL, DDBJ and PDB) databases for class D group II IEPs, using BlastP or TBlastN, with the consensus amino-acid sequence obtained from a multiple sequence alignment (MSA) of domain X and the C- terminal extension of 35 bacterial class D ORFs [18]. Additional class D ORF sequences were identified and the final dataset for phylogenetic analysis included 67 that were #75% identical The alignment used for the phylogenetic analysis of intron ORFs lacking a recognizable D/En region was initially based on the X-domain and C-terminal sequences. It covered a mean of 130 amino acids corresponding to gap-free lengths of 174 unique PLOS ONE \| www.plosone.org January 2013 \| Volume 8 \| Issue 1 \| e55102 January 2013 \| Volume 8 \| Issue 1 \| e55102 2 Group II Introns Lacking the En Domain PLOS ONE \| www.plosone.org 3 January 2013 \| V PLOS ONE \| www.plosone.org 3 January 2013 3 January 2013 \| Volume 8 \| Issue 1 \| e55102 3 PLOS ONE \| www.plosone.org Group II Introns Lacking the En Domain Figure 2. Phylogeny of group II intron ORF classes lacking a recognizable D/En region. (A) Phylogenetic tree based on the alignment of the X-domain and its C-terminal extension. (B) Phylogenetic tree based on the alignment of the RT-domain (0–7). Consensus unrooted trees estimated by ML methods are presented as a radial phylogram. The phylogenetic ORF classes are indicated and the rate of amino-acid substitutions per site is shown at the bottom of each tree. The branches of the major classes are also color-coded to improve visualization of the major clades. Bootstrap results ($75%) are given for each node (PhyML). Posterior probability values ($0.96) for Bayesian analysis of the major phylogenetic ORF classes are also indicated in italics. Introns previously classified as phylogenetic ORF class F are indicated by an asterisk. doi:10.1371/journal.pone.0055102.g002 sequences that were #75% identical (Figure S1). The only exceptions were the class A ORF sequences E.c.I4 and S.f.I1 (99% identity), and the Ni.haI1 (class F) and Ni.haI2 (unclassified) ORFs (99% identity). The larger RT-domain, containing a mean of 257 amino acids, was also used for the phylogenetic analysis of a subset of 123 unique sequences with recognizable RT amino-acid sequences (Figure S2). We included representatives [15] of subclasses B, CL and ML in other analyses. An unrooted tree and 100 boostraps were generated with PhyML [19,20], implemented in Geneious Pro (Biomatters Ltd.), with the WAG amino-acid substitution model. Note that different substitution models (i.e. WAG and LG) delivered trees with the same topology. The GUIDANCE method [21] was used to determine alignment confidence scores and to remove aligned positions considered unreliable according to a given cutoff. GUIDANCE is freely available for use from http://guidance.tau.ac.il. The species delimitation plugin for Genious [22] was used to calculate the sequences that were #75% identical (Figure S1). The only exceptions were the class A ORF sequences E.c.I4 and S.f.I1 (99% identity), and the Ni.haI1 (class F) and Ni.haI2 (unclassified) ORFs (99% identity). The larger RT-domain, containing a mean of 257 amino acids, was also used for the phylogenetic analysis of a subset of 123 unique sequences with recognizable RT amino-acid sequences (Figure S2). We included representatives [15] of subclasses B, CL and ML in other analyses. An unrooted tree and 100 boostraps were generated with PhyML [19,20], implemented in Geneious Pro (Biomatters Ltd.), with the WAG amino-acid substitution model. Note that different substitution models (i.e. Identification of new types of intron-encoded ORF based on the maturase domain and its C-terminal extension As mentioned above, Simon et al. [16] reported a phylogenetic analysis based on the RT/maturase sequence, in which unclas- sified introns were basal to class F, decreasing statistical support for this clade. We investigated the possible relationships between these intron ORFs, by carrying out a phylogenetic analysis of all the class F and unclassified intron ORFs reported in the database, by the maximum likelihood estimation method [19,20]. The phylo- genetic trees (Fig. 1) show that the class F intron M.xa.I1 lies within a well supported group that also contains the unclassified introns Sg.ce.I1, W.e.I5, Ma.sp.I2 and B.th.I3, with bootstrap values of 99% (X-domain and its C-terminal extension, Fig. 1A) and 100% (RT-domain, Fig. 1B). In addition, the related class F introns So.us.I2 and Ge.ur.I1 belong to a cluster that includes another class F intron (P.th.I2) and the unclassified intron c- Ku.st.I1. Moreover, the class F introns D.a.I1 and Ni.ha.I1 cluster with the unclassified intron Ni.ha.I2. The position of the class F intron Pe.ca.I3 is uncertain in both trees. The phylogenetic trees thus indicate a lack of statistical support for the grouping together of class F introns. Phylogenetic analyses were performed on an amino-acid sequence alignment (Figure S1) for domain X and its C-terminal extension for 174 bacterial group II introns, by the maximum likelihood estimation method. The initial set of intron ORFs included all class A, C, D, E, F and unclassified introns reported in the database and the new introns identified in this work. The final dataset used for the analyses contained only selected sequences that were #75% identical (with a few exceptions, as indicated). The estimated phylogenetic tree supports the existence of the previously described classes C, D and E (Fig. 2A), but not that of class F. As previously reported by Simon et al. [16], we found that class E could potentially be split into two clades, E1 and E/E2, but statistical support was strong only for E1, with a 97% bootstrap value and a high posterior probability (1). Ta.sp.I2 appeared to be the most divergent ORF within class E. g Class D, C and other ORFs branched from a common node with a high bootstrap value (79%), but a low posterior probability (0.51). The phylogenetic tree for the domain X and C-terminal amino-acid sequences also suggested the existence of additional varieties of introns, with strong statistical support (bootstrap value .75%) and a high posterior probability ($0.99). Identification of new types of intron-encoded ORF based on the maturase domain and its C-terminal extension One of these new groups (hereafter referred to as g1), supported by a bootstrap value of 93%, contains eight different ORFs, including those of the Ca.ac.I1 and Rh.js.I1 introns previously described as ‘‘unclassi- fied’’. A second group (hereafter referred to as g2) with strong support (99% boostrap) contained 10 different ORFs, including those of the Mx.xa.I1 intron, which had previously been assigned to class F, and the unclassified introns W.e.I5, Ma.sp.I2, Sg.ce.I1 and B.th.I3. A third group (hereafter referred to as g3), with a high posterior probability (1) and high bootstrap values (77%), contained eight introns, including the class F introns D.a.I1 and Ni.ha.I1 and the previously unclassified intron Ni.ha.I2. Finally, a fourth group (hereafter referred to as g4) with strong statistical support (100% boostrap) had eight members, including the class F intron Pe.ca.I3. The relationships between these groups and the major classes remain unclear, because the deeper branches of the tree have yet to be resolved. Notably, maximum likelihood estimation on the basis of a reduced multiple sequence alignment (MSA; 84 informative positions encompassing only the conserved X-domain without the C-terminal extension), with the removal of unreliable columns by the GUIDANCE [21] method (cutoff 0.427 and a guidance alignment score of 0.827309) provided strong statistical support for g4 (96%) and g2 (94%). Likewise, high levels of statistical support were obtained for class D (73%) and class E Relationships between class F and unclassified group II intron-encoded ORFs Relationships between class F and unclassified group II intron-encoded ORFs Group II Introns Lacking the En Domain ORFs included in the final dataset originated from 49 different bacterial species belonging to seven subdivisions of Eubacteria (Acidobacteria, 1; Actinobacteria, 7; Bacteroidetes, 3; Chlorobi, 4; Firmicutes, 9; Proteobacteria, 33 and Thermotogae, 1); four originated from nonculturable environmental bacteria, one from an archaea (Euryarchaeota), one from a nonculturable environ- mental archaea sample and, surprisingly, one was found in a draft plant genome sequence (Ricinus communis). Identification of group II intron boundaries Identification of group II intron boundaries Nucleotide sequences (4–5 kb) containing putative intron- encoded ORFs were analyzed with Mfold for the detection of potential group II intron RNA structures [25]. We searched for intron boundaries with the bioinformatics tool from the intron database [17]. Most of the ORFs presented intron boundaries that were identified with this tool. However, some of the candidate introns of the new varieties described here had no predicted intron boundaries. In these cases, a conserved and, presumably, catalytic domain V was identified, generally just downstream from the ORF sequence. Clustal W alignment of the ORF surrounding the RNA sequence (generally 2.0–2.5 kb in size), together with an exami- nation of putative exon junctions, made it possible to define the boundaries and a consensus group II intron structure (not shown) for these new varieties. We investigated whether the newly identified intron ORFs had an associated ribozyme, by analyzing the secondary structure motifs typical of group II introns. Determinations of the secondary structure of domains I, II and III upstream from the ORFs and of the characteristic downstream domains, V and VI (Table S1 and Table S2), revealed an associated group II intron RNA structure. Moreover, 13 of the 27 newly identified ORFs related to former class F or unclassified introns were full-length, as were 14 of the 41 new class D ORFs. Group II Introns Lacking the En Domain [15] were added (RT 0– 7). The corresponding accession numbers can be obtained from the bacterial group II intron database. A consensus unrooted tree is shown and major ORF classes and the new groups are labeled. Bootstrap results ($75%) are given for each node (PhyML). The arrow indicates the common node of the lineage of group II introns with an En domain in their IEPs. The circle indicates the possible node grouping together class F introns. doi:10.1371/journal.pone.0055102.g003 Figure 3. RT domain-based phylogeny of group II intron ORFs. To the MSA alignment used for Fig. 2B, representatives of classes B: En.fmI3, B.a.I2, En.fm.I1, E.f.I3, B.me.I1, C.d.I1, G.k.I1, and B.c.I5; CL1A: A.v.I1, P.p.I2, E.c.I5, Th.e.I1, B.t.I1, Sh.sp.I1, Ms.b.I1; CL1B: B.me.I2, and B.a.I1; CL2A: Tr.e.I2, G.v.I1, Ns.p.I4, Cs.p.I1, and C.w.I1; CL2B: C.w.I6, Ns.p.I2, and Ns.p.I1; and ML: Ba.fr.I1, B.t.I2, N.a.I2, and L.l.I1 used by Simon et al. [15] were added (RT 0– 7). The corresponding accession numbers can be obtained from the bacterial group II intron database. A consensus unrooted tree is shown and major ORF classes and the new groups are labeled. Bootstrap results ($75%) are given for each node (PhyML). The arrow indicates the common node of the lineage of group II introns with an En domain in their IEPs. The circle indicates the possible node grouping together class F introns. doi:10.1371/journal.pone.0055102.g003 January 2013 \| Volume 8 \| Issue 1 \| e55102 January 2013 \| Volume 8 \| Issue 1 \| e55102 PLOS ONE \| www.plosone.org 4 Comparison of maturase and C-terminal phylogeny with that based on the RT-domain The conserved RT-domain has a larger number of readily alignable characters than the X-domain and its C-terminal extension. It therefore contains more informative positions for phylogenetic analyses. We investigated whether the phylogeny inferred from this domain was consistent with the definition of the new varieties of intron-ORFs described above. Phylogenetic analysis was performed on a subset of 123 unique sequences with a recognizable RT amino-acid sequence, based on the corre- sponding MSA (Figure S2), by the maximum likelihood estimation method and Bayesian inference. In general, the inferred phylogeny was congruent with that obtained with the maturase and C- terminal amino-acid sequences. The estimated phylogenetic tree (Fig. 2B) supported the existence of the previously described classes C, D and E, with 100% bootstrap values and high posterior probabilities (1), but not that of class F. Strong statistical support was also obtained for the new groups g1-4 (posterior probability of 1, boostrap value .80%). Nevertheless, this phylogeny identifies several specific features. Some of the class C intron ORFs (D.p.I1, Al.or.I2 and Pe.th.I1) lie outside the main node, suggesting that they might be divergent ORFs within this class. Intron Sr.me.I2, which remained unclassified in the maturase tree (note that its ORF lacks the C-terminal extension), clustered within g1 in the RT tree. The intron ORFs clustered in g2 contained a characteristic FADD consensus motif, rather than YADD, in the RT5 domain. There was strong support for sister clade status for classes D and E (posterior probability of 1, bootstrap value of 93%). This phylogeny was not altered by the addition to the MSA of intron ORF RT-sequences representative of classes B, CL1, CL2 and ML, all of which have an En-domain in their IEPs. This intron lineage branched from a strongly supported (95% bootstrap) node, suggesting that the En domain was acquired only once, in a common ancestor (Fig. 3). The relationships between the intron ORF classes of the intron lineage lacking the En-domain remain unclear, because the deeper branches of the tree have yet to be resolved. Nevertheless, the phylogeny based on the RT-domain suggests that classes E and D may be sister clades. We therefore obtained no evidence to support the hypothesis that class E introns form part of the lineage of introns with an En domain in their IEPs [15]. Group II Introns Lacking the En Domain Group II Introns Lacking the En Domain introns used to established class F [15,16] belonged to different monophyletic groups, and that only group F members Pe.thI2, So.us.I2 and Ge.ur.I1 appeared to form a distinct clade. Thus, the proposed class F should be reconsidered. The new intron varieties identified here could be classified into four different well supported monophyletic groups (g1-4). Group g1, which contains no introns previously classified as class F, displayed a significant interspecies distance and may constitute a separate ORF-based phylogenetic subclass. By contrast, g2-4 each included group F members and shared a common node with a basal group containing the rest of the former class F introns (Ge.ur.I1, So.us.I2 and Pe.th.I2), and the unclassified intron c-Ku.st.I1. The grouping together of g2-4 and the small basal F group does not seem to be supported by high bootstrap values in the tree. However, we nevertheless propose that class F should be represented by the four former clades until their relationships can be resolved. (82%), whereas the short alignment decreased the bootstrap value for classes C and g1 to 57% and 65%, respectively, and provided no significant support for g3 (not shown). In the short MSA tree, phylogenetic resolution was lower, but nonetheless consistent with the monophyly of g2 and g4. Conclusions We thank the editor and reviewers for comments. We have increased the number of known introns in bacteria and added to knowledge concerning the lineage of group II introns encoding RT/maturase proteins lacking a D/En region. We have identified several new intron ORF varieties and revised the definition of the previously proposed class F. We found that the Figure S2 MSA based on the RT-domain (RT0-7). (TIF) The interspecies distance between the currently recognized classes C, D and E (Fig. 2B) ranged from 2.591 (D:E) to 3.333% (D:C) divergence, and all had significant P(AB) values (P,1025). The interspecies distance of the monophyletic g1-4 ranged from 1.325 (g3:g4) to 2.740% (g1:g2) divergence, but only g1 and g2 have significant P(AB) values, with g1 identified as the most divergent clade. Table S1 Class D group II introns. (XLS) Table S1 Class D group II introns. (XLS) Table S2 New varieties of group II introns lacking the En domain. Table S2 New varieties of group II introns lacking the En domain. (XLS) Comparison of maturase and C-terminal phylogeny with that based on the RT-domain Instead, our data suggest that the En domain was acquired only once, by the common ancestor of classes B, CL and ML. The large number of bacterial genome sequences already available and the continual production of additional sequence data provide the basis for successful analyses of group II intron diversity. We expect even greater intron diversity in bacteria to be detected through future surveys of sequence databases, providing insight into the importance of these catalytic RNAs and retro- elements in bacterial evolution. Author Contributions Conceived and designed the experiments: NT. Performed the experiments: NT. Analyzed the data: NT FMA. Contributed reagents/materials/ analysis tools: NT. Wrote the paper: NT FMA. 1. Sharp PA (1985) On the origin of RNA splicing and introns. Cell 42:397–400. p g p g 2. Cech TR (1986) The generality of self-splicing RNA: relationship to nuclear mRNA splicing. Cell 44:207–210. 6. Dai L, Zimmerly S (2003) ORF-less and reverse-transcriptase-encoding group II introns in archaebacteria, with a pattern of homing into related group II intron ORFs. RNA 9:14–19. Updated compilation of bacterial group II intron- encoded ORFs lacking the endonuclease domain Bacterial group II intron ORF phylogenetic classes containing only reverse transcriptase/maturase open reading frames, with no recognizable D/En region, belong to class A, which had only three almost identical (99% identity) members, class C, class E and class D. Moreover, class E was split into two clades, E1 and E2 [16], whereas other ORFs in the intron database were classified simply as ‘‘class E’’. We therefore carried out a comprehensive phylogenetic analysis of intron ORFs and investigated the phylogeny of the proposed class F and the other unclassified introns further, by generating a larger dataset of related ORFs, as described in the materials and methods. In this analysis, we identified 27 new ORFs (Table S2) not reported in the intron database, that were related to the proposed class F and unclassified introns; only 14 of these ORFs had a complete RT-X sequence. The 50 ORFs included in the final dataset originated from 37 different bacterial species and six subdivisions of Eubacteria (Acidobacteria, 1; Actinobacteria, 16; Bacteroidetes, 1; Firmicutes, 10; Proteobacteria, 20 and Verrumicrobia, 1). One ORF was found in a nonculturable environmental archaea sample. We also updated the compilation of class D intron-encoded ORFs and identified 41 not reported in the intron database, 18 of which had a complete RT-X sequence (Table S1). The 67 class D January 2013 \| Volume 8 \| Issue 1 \| e55102 PLOS ONE \| www.plosone.org 5 3. Cavalier-Smith T (1991) Intron phylogeny: a new hypothesis. Trends Genet 7:145–148. 4. Eickbush TH (1994) Origins and evolutionary relationships of retroelements. In The Evolutionary Biology of Viruses. Edited by Morse SS. New York: Raven Press; 121–157. 5. Michel F, Ferat JL (1995) Structure and activities of group II introns. Annu Rev Biochem 64: 435–461. 1. Sharp PA (1985) On the origin of RNA splicing and introns. 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Molina-Sa´nchez MD, Martı´nez-Abarca F, Toro N (2010) Structural features in the C-terminal region of the Sinorhizobium meliloti RmInt1 group II intron- encoded protein contribute to its maturase and intron DNA-insertion function. FEBS J 277:244–254. j p g 10. Michel F, Costa M, Westhof E (2009) The ribozyme core of group II introns: a structure in want of partners. Trends Biochem Sci 34:189–199. j p g 10. Michel F, Costa M, Westhof E (2009) The ribozyme core of gro structure in want of partners. Trends Biochem Sci 34:189–199 19. Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52:696–704. 11. Mohr G, Perlman PS, Lambowitz AM (1993) Evolutionary relationships among group II intron-encoded proteins and identification of a conserved domain that may be related to maturase function. Nucleic Acids Res 21: 4991–4997. 20. Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, et al. 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Drummond AJ, Ashton B, Buxton S (2010) Geneious v5.1, Available: http:// www.geneious.com. II intron ORFs. Nucleic Acids Res 29:1238–1250. 23. Rosenberg NA (2007) Statistical tests for taxonomic distinctiveness from observations of monophyly. Evolution 61(2):317–23. 15. References 5. Michel F, Ferat JL (1995) Structure and activities of group II introns. Annu Rev Biochem 64: 435–461. 4. Eickbush TH (1994) Origins and evolutionary relationships of retroelements. In The Evolutionary Biology of Viruses. Edited by Morse SS. New York: Raven Press; 121–157. 7. Toro N (2003) Bacteria and archaea group II introns: additional mobile genetic elements in the environment. Environ Microbiol 5: 143–151. January 2013 \| Volume 8 \| Issue 1 \| e55102 PLOS ONE \| www.plosone.org 6 Group II Introns Lacking the En Domain Simon DM, Kelchner SA, Zimmerly S (2009) A broadscale phylogenetic analysis of group II intron RNAs and intron-encoded reverse transcriptases. Mol Biol Evol 26: 2795–2808. p y y ( ) 24. 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https://openalex.org/W4319997520	https://www.light-am.com/article/pdf/preview/LAM2022050014.pdf	unk	null	Iterative projection meets sparsity regularization: towards practical single-shot quantitative phase imaging with in-line holography	Deleted Journal	2,023	cc-by	13,103	Abstract Holography provides access to the optical phase. The emerging compressive phase retrieval approach can achieve in-line holographic imaging beyond the information-theoretic limit or even from a single shot by exploring the signal priors. However, iterative projection methods based on physical knowledge of the wavefield suffer from poor imaging quality, whereas the regularization techniques sacrifice robustness for fidelity. In this work, we present a unified compressive phase retrieval framework for in-line holography that encapsulates the unique advantages of both physical constraints and sparsity priors. In particular, a constrained complex total variation (CCTV) regularizer is introduced that explores the well-known absorption and support constraints together with sparsity in the gradient domain, enabling practical high-quality in-line holographic imaging from a single intensity image. We developed efficient solvers based on the proximal gradient method for the non-smooth regularized inverse problem and the corresponding denoising subproblem. Theoretical analyses further guarantee the convergence of the algorithms with prespecified parameters, obviating the need for manual parameter tuning. As both simulated and optical experiments demonstrate, the proposed CCTV model can characterize complex natural scenes while utilizing physically tractable constraints for quality enhancement. This new compressive phase retrieval approach can be extended, with minor adjustments, to various imaging configurations, sparsifying operators, and physical knowledge. It may cast new light on both theoretical and empirical studies. Keywords: Phase retrieval, Quantitative phase imaging, Compressive sensing, Digital holography Article Article Open Access Open Access © The Author(s) 2023 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article′s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article′s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Iterative projection meets sparsity regularization: towards practical single-shot quantitative phase imaging with in-line holography Yunhui Gao 1 and Liangcai Cao 1,* Correspondence: Liangcai Cao (clc@tsinghua.edu.cn) 1State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instruments, Tsinghua University, Beijing 100084, China Official journal of the JHL 2689-9620 www.light-am.com Official journal of the JHL 2689-9620 www.light-am.com Gao et al. Light: Advanced Manufacturing (2023)4:6 https://doi.org/10.37188/lam.2023.006 Official journal of the JHL 2689-9620 www.light-am.com Introduction detection devices can directly record only the average incident power. The loss of phase information poses unique challenges for the numerical inversion of the imaging process, leading to the so-called “twin-image” effect that severely degrades the imaging quality 4,5. As a result, phase retrieval, or the recovery of the phase from intensity-only measurements, remains a longstanding issue and lies at the heart of digital holography and many related optical imaging techniques 6. In recent years, phase retrieval has The phase of an optical field contains various sources of information. Holography, invented by Gabor in 1947 1, provides access to phase information and has been used as a powerful imaging technique 2,3. At the optical frequency, however, electromagnetic waves oscillate so fast that the Page 2 of 17 Gao et al. Light: Advanced Manufacturing (2023)4:6 regained increasing research interests following the development of computational imaging, which leverages advancements in signal processing theory, numerical algorithms, hardware design, and computation power 7. superior imaging performance, they are highly dependent on the scene and may require manual fine-tuning to achieve satisfactory results. In pursuit of a compressive phase retrieval method for in-line holography that features both robustness and high fidelity, we propose a novel computational framework that encapsulates physical constraints and sparsity priors as a unified optimization problem. Specifically, we consider the well-known absorption and support constraints and gradient-domain sparsity, leading to a constrained complex total variation (CCTV) model. This is because TV- regularized problems can be solved using computationally efficient algorithms and are thus highly scalable to large- scale problems 94,95. The piece-wise smoothness underlying the TV model can also characterize multidimensional data such as the three-dimensional refractive index distribution, offering good generalizability to various applications 96−99. Furthermore, TV-regularized inversion algorithms offer mathematically tractable convergence behaviors, which is important for both theoretical soundness and practical implementation. Existing phase retrieval methods for in-line holography can be broadly classified into two categories, namely diversity phase retrieval and compressive phase retrieval. The former approach requires taking multiple images with some data diversity that translate phase information into intensity distribution. This can be achieved, for example, by varying the imaging distance 8−10, illumination wavelength 11−14, probe position 15−17, illumination angle 18−20, and modulation pattern 21−24. While this approach can achieve quantitative phase imaging with increased data diversity and redundancy, it sacrifices the temporal resolution, system cost, and complexity. Introduction As an alternative, compressive phase retrieval takes a different path by exploring additional prior knowledge of the wavefield, which could potentially achieve phase recovery beyond the information-theoretic limit or even from a single shot. Historically, early works tackled the phase ambiguity problem by utilizing simple yet effective physical constraints such as non-negativity 25,26, support 27−29, absorption 30, histogram 31, and atomicity 32. These constraints are typically enforced via projections in the complex Euclidean space, and the corresponding phase retrieval algorithms proceed by projecting the estimated wavefield back in an iterative manner 33,34. Since the seminal Gerchberg-Saxton 35 and Fienup’s error-reduction algorithms 36, iterative projection algorithms have played a pivotal role in phase retrieval for decades 37−39. However, this classical approach relies on carefully handcrafted physical constraints, which may fail to fully extract the inherent features of complex scenes under limited measurements, particularly in the case of single-shot phase retrieval 40. Consequently, it has mostly been demonstrated for simple objects. The quantitative phase analysis of complex samples, such as dense biological specimens, remains a challenging task. Based on the CCTV model, compressive phase retrieval is formulated as a regularized inverse problem and is solved via an accelerated proximal gradient algorithm together with an efficient denoiser for the proximal update. Unlike conventional methods that rely on a heuristic basis, we provide a rigorous mathematical analysis that guarantees the convergence of the proposed algorithms using prespecified parameters. We experimentally demonstrated quantitative phase imaging of various samples from a single in-line hologram, illustrating the effectiveness of the CCTV model in characterizing complex real-world scenes while leveraging physically tractable constraints for quality enhancement. We further elaborate on algorithmic behaviors based on extensive numerical studies, corroborating the theoretical results while providing general guidance for practitioners. Results The concept of compressive phase retrieval was formally established by an explosion of theoretical and empirical studies following the development of compressed sensing 41−43. Sparsity priors in various domains such as the spatial 44−49, gradient 50−61, wavelet 62 and other domains 63,64 or with a dictionary-learned transform 65−67 have been demonstrated as effective regularizers for phase recovery. More recently, implicit image priors from advanced denoisers such as BM3D 68−73 or represented by deep neural networks 74−93 have also been studied in the context of phase retrieval, resulting in improved reconstruction quality. Although the regularization techniques generally yield Phase retrieval via CCTV For electromagnetic waves at optical frequencies, sensors can only respond to the average power of the incident wavefield. Thus, phase information, i.e., the complex argument of the signal, is lost during the measurement process. The forward model can be abstractly expressed, regardless of the imaging configuration, in a vectorized form as y = \|Ax\| (1) (1) y ∈Rm A ∈Cm×n where is the observed measurement, is the y ∈Rm A ∈Cm×n where is the observed measurement, is the Page 3 of 17 Page 3 of 17 Gao et al. Light: Advanced Manufacturing (2023)4:6 Generally, the sampling matrix may involve various linear physical processes such as probe illumination, free- space propagation and mask modulation, all of which are determined by the optical configuration of the imaging system. In this work, we consider the lensless in-line holographic imaging model for illustration, which is shown in Fig. 1a. The object is illuminated by a coherent plane wave, such that the retrieved complex field directly reveals the absorption and phase shift introduced by the object. An x ∈Cn x y sampling matrix, and is the underlying complex signal that represents the transmission of the object. In the vectorized formulation, column vectors and are raster- scanned from the corresponding two-dimensional complex object transmission function and hologram amplitude, respectively. It is important to note that, the purpose of vectorization is to simplify the algorithm derivation and notations. Vectorization is not required when implementing the algorithm in practice. 0 1.2 (rad) Proximal gradient method Circular convolution via FFT Image cropping θ Sensor area Complex object x Measurement y Sensor Object Lens Spatial filter Coherent source Data fidelity F(x) 1/2 \|\| \|Ax\| − y \|\|2 2 Physical constraint only Gradient sparsity only Physical constraint + gradient sparsity Amplitude Phase PC y = \|Ax\| Gradient update x ← x − γÑxF(x) Proximal update x ← proxγR(x) Iterative denoiser Amplitude Phase 0 Re Im Horizontal Vertical Enforce constraints Projection onto set C Support 0 0 1 0 Absorption Phase 0 Amplitude 1 0 Objective: Regularization Finite differences Dx Σ minimize F(x) x R(x) R(x) λ \|\| Dx \|\|1 IC(x) = + + a Phase imaging system b Mathematical formulation e Algorithmic framework f Retrieved phase c Physical constraint d Gradient sparsity Xi,j Xi,j+1 or Xi+1,j Forward model Signal priors = θ Fig. 1 Schematic of the proposed compressive phase retrieval framework. Phase retrieval via CCTV a The optical configuration of an in-line holographic imaging system. b The angular spectrum diffraction model adopted for numerical calculation. The diffraction angle and the corresponding convolution kernel size are determined by the sampling frequency (or, equivalently, the sensor pixel size). c Physical knowledge, such as object support or absorption, is often enforced as hard constraints via projection operations. d The complex TV regularizer promotes sparsity in both the gradients of amplitude and phase. e We combine the forward model and signal priors into an optimization problem, which is solved via the proximal gradient method. An iterative denoiser is introduced for the proximal update step. f Retrieved phase from experimental data. Combining physical constraints and gradient sparsity yields the best performance. Circular convolution via FFT Image cropping θ Sensor area Complex object x Measurement y Sensor Object Lens Spatial filter Coherent source Amplitude Phase PC y = \|Ax\| a Phase imaging system b Mathematical formulation Forward model Enforce constraints Projection onto set C Support 0 0 1 0 Absorption Phase 0 Amplitude 1 0 c Physical constraint Signal priors 1 0 Lens PC Circular convolution via FFT Image cropping θ Sensor area Complex object x Measurement y Amplitude Phase y = \|Ax\| b Mathematical formulation b Mathematical formulation b Amplitude Phase 0 Re Im Horizontal Vertical Finite differences Dx Σ d Gradient sparsity Xi,j Xi,j+1 or Xi+1,j d Gradient sparsity Measurement y Complex object x 0 1.2 (rad) Proximal gradient method Data fidelity F(x) 1/2 \|\| \|Ax\| − y \|\|2 2 Physical constraint only Gradient sparsity only Physical constraint + gradient sparsity Gradient update x ← x − γÑxF(x) Proximal update x ← proxγR(x) Iterative denoiser Objective: Regularization minimize F(x) x R(x) R(x) λ \|\| Dx \|\|1 IC(x) = + + e Algorithmic framework f Retrieved phase = θ Fig. 1 Schematic of the proposed compressive phase retrieval framework. a The optical configuration of an in-line holographic imaging system. b The angular spectrum diffraction model adopted for numerical calculation. The diffraction angle and the corresponding convolution kernel size are determined by the sampling frequency (or, equivalently, the sensor pixel size). c Physical knowledge, such as object support or absorption, is often enforced as hard constraints via projection operations. d The complex TV regularizer promotes sparsity in both the gradients of amplitude and phase. Phase retrieval via CCTV As shown in Fig. 1d, the complex TV reveals both the amplitude and phase variations between adjacent sampling points of the object transmission function and thus serves as a good sparsifying transform for complex-valued images 113. Apart from the complex TV, we introduce a second regularizer , which is an indicator function of the constraint set . This is motivated by conventional iterative projection (IP) phase retrieval algorithms, where phase retrieval is formulated as a feasibility problem, and the unknown field distribution is assumed to belong to certain physical constraint sets. The IP algorithms typically proceed by projecting the estimated wavefield onto constraint sets in an iterative manner 37,38. Here, we consider the support and absorption constraints in particular because they are commonly adopted for reconstructing complex objects, and their corresponding constraint sets are both convex. Fig. 1c presents a conceptual illustration of these physical constraints, and the mathematical definitions are presented in Table 1. The proposed regularizer comprises a complex TV and a hard physical constraint and is thus termed the constrained complex total variation regularizer. As a special case, when , CCTV reduces to the complex TV function without physical constraints, and when , it reduces to Phase reconstruction from a single in-line hologram is inherently challenging due to the existence of ambiguous solutions. Therefore, compressive phase retrieval takes an inverse problem approach that utilizes regularization techniques to ensure well-posedness. In this work, we introduce the integration of a complex TV and a hard constraint as the regularizer, leading to an optimization problem in the following form: min x 1 2 ∥\|Ax\|−y∥2 2 \| {z } F(x) +λ∥Dx∥1 + IC(x) \| {z } R(x) (3) (3) ∥·∥p ℓp F(x) R(x) where denotes the vector norm or the corresponding matrix norm. is the data-fidelity term and is the regularization term. The fidelity term ensures that the solution does not deviate significantly from the forward model in Eq. 1. Meanwhile, the regularization term introduces additional prior knowledge of the wavefield to improve the reconstruction quality further. F(x) F(x) = (1/2)∥\|Ax\|2 −y2∥2 2 F(x) = (1/2)∥\|Ax\|−y∥2 2 There are various choices for the fidelity function in the phase retrieval literature, such as the intensity-based formulation 104−106 and the amplitude-based formulation 107−109. The main reasons for choosing the amplitude-based residual here are threefold. Phase retrieval via CCTV e We combine the forward model and signal priors into an optimization problem, which is solved via the proximal gradient method. An iterative denoiser is introduced for the proximal update step. f Retrieved phase from experimental data. Combining physical constraints and gradient sparsity yields the best performance. Proximal update x ← proxγR(x) Iterative denoiser Regularization R(x) λ \|\| Dx \|\|1 IC(x) = + Iterative denoiser Proximal gradient method f Retrieved phase f Retrieved phase Physical constraint only p 0 1.2 (rad) Physical constraint + gradient sparsity Gradient sparsity only Gradient sparsity only Physical constraint only Physical constraint + gradient sparsity θ Fig. 1 Schematic of the proposed compressive phase retrieval framework. a The optical configuration of an in-line holographic imaging system. b The angular spectrum diffraction model adopted for numerical calculation. The diffraction angle and the corresponding convolution kernel size are determined by the sampling frequency (or, equivalently, the sensor pixel size). c Physical knowledge, such as object support or absorption, is often enforced as hard constraints via projection operations. d The complex TV regularizer promotes sparsity in both the gradients of amplitude and phase. e We combine the forward model and signal priors into an optimization problem, which is solved via the proximal gradient method. An iterative denoiser is introduced for the proximal update step. f Retrieved phase from experimental data. Combining physical constraints and gradient sparsity yields the best performance. Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 4 of 17 simplifying the implementation. Third, many empirical studies have observed faster convergence when minimizing amplitude-based fidelity functions 111,112. imaging sensor was placed close to the object, and the in- line hologram was recorded. Such a simple optical configuration enables low-cost and portable system design for various imaging applications 100,101. The physical process involves only free-space propagation of the wavefield, which is calculated using the angular spectrum method 102. Therefore, the sampling matrix of our imaging model can be expressed as R(x) λ > 0 The proposed regularization function comprises two separate terms. The first term is the sparsity regularizer with the parameter . Phase retrieval via CCTV Specifically, we considered the anisotropic complex TV seminorm: ∥Dx∥1 =TV(X) = nξ−1 ∑ i=1 nυ ∑ j=1 \|Xi+1,j−Xi,j\|+ nξ ∑ i=1 nυ−1 ∑ j=1 \|Xi,j+1−Xi,j\| (4) A = CQ (2) (2) (4) Q ∈Cn×n C ∈Rm×n m < n where denotes the free-space propagation operator calculated as a circular convolution with the Fresnel kernel via fast Fourier transforms (FFTs), denotes an image cropping operation to model the finite size of the imaging sensor 103. Notice that we have , that is, the object dimension is larger than the pixel number of the sensor. This is because out-of-field objects may also contribute to the recorded image owing to the linear convolution effect 54. This process is illustrated in Fig. 1b. X ∈Cnξ×nυ nξnυ = n x D ∈C2n×n x IC(x) C R(x) C = Cn λ = 0 where with denotes the non-vectorized two-dimensional image corresponding to . denotes the spatial finite-difference operator for the vectorized along the two directions. As shown in Fig. 1d, the complex TV reveals both the amplitude and phase variations between adjacent sampling points of the object transmission function and thus serves as a good sparsifying transform for complex-valued images 113. Apart from the complex TV, we introduce a second regularizer , which is an indicator function of the constraint set . This is motivated by conventional iterative projection (IP) phase retrieval algorithms, where phase retrieval is formulated as a feasibility problem, and the unknown field distribution is assumed to belong to certain physical constraint sets. The IP algorithms typically proceed by projecting the estimated wavefield onto constraint sets in an iterative manner 37,38. Here, we consider the support and absorption constraints in particular because they are commonly adopted for reconstructing complex objects, and their corresponding constraint sets are both convex. Fig. 1c presents a conceptual illustration of these physical constraints, and the mathematical definitions are presented in Table 1. The proposed regularizer comprises a complex TV and a hard physical constraint and is thus termed the constrained complex total variation regularizer. As a special case, when , CCTV reduces to the complex TV function without physical constraints, and when , it reduces to X ∈Cnξ×nυ nξnυ = n x D ∈C2n×n x IC(x) C R(x) C = Cn λ = 0 where with denotes the non-vectorized two-dimensional image corresponding to . denotes the spatial finite-difference operator for the vectorized along the two directions. ⊙ 1 The symbol denotes the element-wise (Hadamard) multi- plication operator. 0 and 1 denote the zero vector and all-ones vector, respectively. P denotes the set of pixels outside the support region, and 1P denotes the indicator vector of P. Quantitative phase imaging applications We built a proof-of-principle in-line holographic imaging system for optical experiments, as shown in Fig. 2a. A 660 nm diode-pumped solid-state laser (Cobolt Flamenco 300) was used as the source for coherent illumination. The optical beam generated by the laser passes through a spatial filter that comprises an objective lens and a pinhole at the back focal plane. The filtered wave was then collimated using a convex lens to illuminate CMOS sensor Object Aperture Lens Spatial filter Laser Mirror Polarizer ND filter a b d f g e c 0 3 0 4 (rad) (μm) 1 mm 1 mm 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 0 0.2 0.4 Location (mm) Location (mm) 0 4.5 0.6 0 0.45 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 0 0.5 1.0 2.5 1.5 0.5 0 2.0 0 5 4 3 2 1 1.0 ξ (mm) ξ (mm) ξ (mm) ξ (mm) ν (mm) ν (mm) ν (mm) 1 2 3 1 2 0 0 3 1 2 3 2 1 0 0.6 00 3 0 0 0.4 4 2 0 2 1 ξ ν Height (μm) Height (μm) 0 0.2 0.4 0.6 Height (μm) Height (μm) ν (mm) Fig. 2 Experimental characterization of a fabricated phase plate. a Optical setup of the in-line holographic imaging system. b and c show the captured in-line holograms of two transparent patterns. The inset shows an image of the phase plate. d and e show the reconstructed phase distribution from b and c, respectively. The white solid rectangle in d indicates the sensor’s field of view. f and g visualize the rendered surface profiles and the cross-sectional height profiles of d and e, respectively. The white dashed rectangles indicate the enlarged areas, and the yellow dashed lines indicate the cross-sections. The same applies to other figures. A Spatial filter Laser Mirror Polarizer ND filter a CMOS sensor Object Aperture Lens Spatial filter Laser Mirror Polarizer ND filter a c 0 3 CMOS sensor Object Aperture Lens b b c Lens 0 0 e d 1 mm ξ ν e 0 4 (rad) 1 mm f 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 0 0.2 0.4 Location (mm) 0 4.5 0.6 2.5 1.5 0.5 0 2.0 0 5 4 3 2 1 1.0 ξ (mm) ξ (mm) ν (mm) ν (mm) 0 0 0.4 4 2 0 2 1 Height (μm) Height (μm) Fig. Phase retrieval via CCTV First, from a physical perspective, it is closely related to the maximum likelihood estimation under Poisson noise, which captures the noise statistics more accurately than the Gaussian noise model 110. Second, on the practitioner’s side, using the amplitude- based formulation ensures the convergence of the iterative algorithms with a prespecified algorithm step size, obviating the need for complicated line searches and Table 1 Physical constraints for phase retrieval Constraint Definition of C x′ = PC(x) Projection Support x⊙1P = 0 x′ i = { 0, i ∈P xi, i < P Absorption \|x\| ≤1 x′ i = { xi/\|xi\|, \|xi\| > 1 xi, \|xi\| ≤1 ⊙ 1 The symbol denotes the element-wise (Hadamard) multi- plication operator. 0 and 1 denote the zero vector and all-ones vector, respectively. P denotes the set of pixels outside the support region, and 1P denotes the indicator vector of P. Table 1 Physical constraints for phase retrieval Table 1 Physical constraints for phase retrieval Constraint Definition of C x′ = PC(x) Projection Support x⊙1P = 0 x′ i = { 0, i ∈P xi, i < P Absorption \|x\| ≤1 x′ i = { xi/\|xi\|, \|xi\| > 1 xi, \|xi\| ≤1 ⊙ 1 The symbol denotes the element-wise (Hadamard) multi- plication operator. 0 and 1 denote the zero vector and all-ones vector, respectively. P denotes the set of pixels outside the support region, and 1P denotes the indicator vector of P. Page 5 of 17 Gao et al. Light: Advanced Manufacturing (2023)4:6 the classical problem of phase retrieval from a modulus measurement and a physical constraint. the object. The diffraction pattern was recorded using a CMOS sensor (QHY174GPS, pixel size 5.86 μm, resolution 1920×1200) which was placed a few millimeters away from the object. The captured raw intensity image was then digitally processed using a personal computer for holographic reconstruction. In all optical experiments, we consider only the absorption constraint because the objects have extended field distributions and thus the support constraint is inapplicable. Before running the iterative phase retrieval algorithm, a preprocessing step is required to determine the imaging distance, which can be achieved by either manual tuning or using some autofocusing Quantitative phase imaging applications 2 Experimental characterization of a fabricated phase plate. a Optical setup of the in-line holographic imaging system. b and c show the captured in-line holograms of two transparent patterns. The inset shows an image of the phase plate. d and e show the reconstructed phase distribution from b and c, respectively. The white solid rectangle in d indicates the sensor’s field of view. f and g visualize the rendered surface profiles and the cross-sectional height profiles of d and e, respectively. The white dashed rectangles indicate the enlarged areas, and the yellow dashed lines indicate the cross-sections. The same applies to other figures. Page 6 of 17 Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 6 of 17 We imaged a muscle tissue section to validate the proposed method on complex objects, as shown in Fig. 3. The twin image was effectively eliminated from the retrieved amplitude and phase, which visually demonstrated the high-fidelity imaging performance of the method. The retrieved phase reveals thickness variations inside the tissue slice, facilitating label-free and low-cost clinical studies or diagnoses 120,121. algorithms 114−116. Each captured intensity image is normalized by the background image which is obtained using the same experimental conditions, only with the object removed. This calibration process ensures the validity of the absorption constraint 39. We demonstrated the quantitative phase imaging capability of the proposed compressive phase retrieval method for real-world objects. Fig. 2 shows the imaging results of a phase plate, which was fabricated by etching multiple self-designed binary patterns onto a JGS1 quartz glass substrate. The retrieved phases of the spiral pattern and Fresnel zone plate are shown in Fig. 2d, e, respectively. Smooth staircase profiles with sharp edges can be accurately retrieved. Artifacts on the surface were preserved, indicating a high spatial resolution. Given that the object has a uniform refractive index distribution and that the fabricated features lie only on the surface of the substrate, the surface profiles can be directly calculated and characterized with knowledge of the medium refractive index, as shown in Fig. 2f, g. It is important to note that recovering the low-frequency phase is inherently challenging for the in-line configuration because the weak phase transfer function diminishes to zero together with the spatial frequency 117. Hence, slowly varying phase structures hardly contribute to the intensity variations at the sensor plane, rendering them particularly difficult to recover. Quantitative phase imaging applications This has been the main drawback for in-line holography compared to off-axis holography 118,119. Our findings suggest that the CCTV regularizer may be a competitive solution for this problem. In addition, because we have addressed the issue of the circular convolution model, the retrieved image is free from boundary artifacts, and the object distribution outside the sensor area can be partially recovered, as shown in Fig. 2d. Ablation experiments λ = 0 C = Cn C We conducted ablation studies based on experimental data to quantify the performance improvements obtained by the physical constraints and the complex TV regularizer respectively. For comparison, we implemented compressive phase retrieval algorithms using only physical constraints or gradient sparsity. The former, referred to as the IP method, can be viewed as an accelerated variant of the conventional iterative projection algorithm. This was implemented by setting the regularization parameter . The algorithm proceeds by iteratively minimizing the data- fidelity function with a gradient descent step and enforcing physical constraints with a projection step. The latter, referred to as the CTV method, uses only the complex TV regularizer without any physical constraints. It is implemented by defining the constraint set and thus the projection onto set is reduced to identity mapping. Both the IP and CCTV methods can utilize various physical constraints. To better differentiate among different methods, we use the letter ‘A’ to represent the method that uses the absorption constraint, the letter ‘S’ for the support constraint, and ‘AS’ for both constraints. For example, ‘IP- A’ represents an iterative projection algorithm that uses only the absorption constraint. We first performed phase imaging of a quantitative 0 3 0 1.2 0 1.2 (rad) a b c 1 mm Fig. 3 Holographic imaging results of a muscle tissue section. a Experimentally recorded intensity image. The inset is an image of the sample slide. b and c are the retrieved amplitude and phase, respectively. b 1.2 0 0 Fig. 3 Holographic imaging results of a muscle tissue section. a Experimentally recorded intensity image. The inset is an image of the sample slide. b and c are the retrieved amplitude and phase, respectively. Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 7 of 17 phase microscopy target (Benchmark Technologies) using three methods, namely IP-A, CTV, and CCTV-A, as shown in Fig. 4. Note that the object has an extended distribution and thus the support constraint is inapplicable. The IP algorithm fails to eliminate the twin-image artifact completely, and the phase structures are not clearly revealed, which implies that the physical constraint alone is insufficient to suppress the phase ambiguities. The CCTV method achieved better imaging quality than CTV, as illustrated by visual and quantitative comparisons. The ground-truth phase values were calculated using the structures’ height and medium refractive index, which are both provided by the manufacturer. Ablation experiments Furthermore, as indicated by a qualitative visual comparison, the IP method suffers from high reconstruction noise, and the CCTV model improves the performance of CTV by suppressing artifacts with no loss of spatial resolution. retrieved using the IP-A, CTV, and CCTV-A methods were 0.880, 0.463, and 0.418 rad, respectively. In addition, the finest resolved features of the USAF 1951 resolution target are highlighted. Group 6, Element 3 is resolved by all three methods, indicating a spatial resolution of 80.6 lines/mm, which agrees with the maximum resolution of 85.3 lines/mm of the sensor. All three methods achieve the same sampling-limited resolution, which implies that the TV- based regularizer does not sacrifice the spatial resolution for quality enhancement. The resolution can be further improved beyond the Nyquist sampling limit, however, if one takes into account the down-sampling effect of the sensor pixels 112,122−126. Ablation experiments The root-mean-square errors (RMSEs) of the cross-sectional phase profiles 4 3 2 1 0 1 0 2 3 a c d f e b 0 0.4 Angle (degree) Phase (rad) 0 20 40 60 80 100 120 140 160 0 0.5 1.0 1.5 0 1.8 (rad) (μm) 0 1.8 (rad) (μm) CTV CCTV-A IP-A 1 2 3 1 2 0 4 0 3 0 0.4 0.4 0.2 0 0 0.2 0.4 0 0.3 0 0.2 0.4 0 0.4 ν (mm) ν (mm) ν (mm) ν (mm) 0 0.4 0 0.4 0 0.3 ν (mm) ν (mm) Height (μm) ξ (mm) ξ (mm) ξ (mm) 0 0.2 0.4 ξ (mm) 0 0.2 0.4 ξ (mm) ξ (mm) ν ξ 500 μm Height (μm) 0.4 0.2 0 0 0.2 0.4 0 0.3 ν (mm) ξ (mm) Height (μm) 0.4 0.2 0 0 0.2 0.4 0 0.3 ν (mm) ξ (mm) Height (μm) Fig. 4 Experimental comparison of different phase retrieval algorithms with a quantitative phase resolution target. a Full field-of-view phase reconstruction by the proposed CCTV-A method. b The calculated surface profile corresponding to a. c, d and e show the retrieved phase and height profiles by the IP-A, CTV, and CCTV-A methods, respectively. f The cross-sectional phase profiles, where the dashed line indicates the calculated theoretical phase value. a ν ξ 500 μm b a 4 3 2 1 0 1 0 2 3 b 1 2 3 1 2 0 4 0 3 0 0.4 ν (mm) ν (mm) Height (μm) ξ (mm) ξ (mm) m Height (μm) 0 1.8 (rad) 0 0.4 c d e 0 0.3 f Fig. 4 Experimental comparison of different phase retrieval algorithms with a quantitative phase resolution target. a Full field-of-view phase reconstruction by the proposed CCTV-A method. b The calculated surface profile corresponding to a. c, d and e show the retrieved phase and height profiles by the IP-A, CTV, and CCTV-A methods, respectively. f The cross-sectional phase profiles, where the dashed line indicates the calculated theoretical phase value. Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 8 of 17 Quantitatively, the RMSEs of the retrieved amplitude profiles 1/2 in Fig. 5d are 0.331/0.338, 0.240/0.239, 0.238/0.232 for the IP-A, CTV and CCTV-A methods, respectively. The proximal gradient framework F(x) R(x) F(x) In the regularized inverse problem of Eq. 3, the fidelity function is a (almost everywhere) differentiable function whereas the regularizer is a non- differentiable function. Because we are dealing with real- valued functions over complex-valued variables, CR- calculus is adopted 127. The gradient of is given by 112 Similar improvements in the CCTV method over the IP and CTV methods were also observed in the imaging results of an amplitude object. Fig. 5 compares the amplitudes of a positive 1951 USAF test target (Thorlabs, Inc.) retrieved by the IP-A, CTV, and CCTV-A methods. a d e d utilp m A Profile 1 Profile 2 0 1.2 b c 1.0 0.5 IP-A IP-A CTV CTV CCTV-A CCTV-A 1 mm Profile 1 Profile 1 Profile 2 Profile 2 40 80 120 140 0 20 60 100 160 Pixel index 0 50 100 150 200 250 Pixel index Fig. 5 Experimental comparison of different phase retrieval algorithms with an amplitude resolution target. a, b and c show the retrieved amplitude using the IP-A, CTV, and CCTV-A algorithms, respectively. The inset indicates the finest structures resolved by the corresponding method. Below are the corresponding enlarged areas. d Cross-sectional profiles. a 1 mm a Fig. 5 Experimental comparison of different phase retrieval algorithms with an amplitude resolution target. a, b and c show the retrieved amplitude using the IP-A, CTV, and CCTV-A algorithms, respectively. The inset indicates the finest structures resolved by the corresponding method. Below are the corresponding enlarged areas. d Cross-sectional profiles. Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 9 of 17 ∇xF(x) = 1 2 AHdiag ( Ax \|Ax\| ) (\|Ax\|−y) (5) dual, which is given by (5) (7) min w∈S { G(w) ≡−∥HC(v−γDHw)∥2 2 +∥v−γDHw∥2 2 } (·)H Ax where denote the conjugate transpose (Hermitian) operator. Non-smoothness only occurs when has zero entries, which, as we will show below, does not affect the algorithm’s convergence behavior. w ∈C2n HC(x) def= x−PC(x) PC C S w ∈C2n where denotes the dual variable, with representing the Euclidean projection operator on , and denotes the convex set which contains all that satisfy R(x) R(x) proxγR The difficulty in minimizing the objective function mainly arises from the non-differentiability of . The proximal gradient method provides a general framework for solving composite non-smooth optimization problems in the form of Eq. Convergence analysis x(0) γ βt = t/(t +3) T Input: Initial guess , step size , , and iteration number . γ η In the above subsections, we introduce the proximal gradient algorithm and the corresponding denoising algorithm for the proximal update. We now show that the algorithm parameters, namely the step sizes and , can be predetermined based on the measurement scheme and the sparsifying transform. T number . u(0) = x(0) 1: t = 1,2,...,T 2: for do v(t) = u(t−1) −γ∇uF(u(t−1)) ▷ 3: Gradient update x(t) = proxγR(v(t)) ▷ 4: Proximal update via Alg. 2 u(t) = x(t) +βt(x(t) −x(t−1)) 5: 6: end for 6: end for Considering the nonconvexity of the problem of Eq. 3, we present a weaker result by showing the convergence of the basic (non-accelerated) proximal gradient algorithm, which is summarized by the following theorem. The detailed proof is provided in the Supplementary Document. Furthermore, extensive numerical studies have demonstrated that the accelerated algorithm converges following the same step size selection rule. The proximal gradient framework Through numerical studies, we empirically demonstrate that it also helps to speed up convergence for the nonconvex phase retrieval problem. ∇wG(w) = −γDPC(v−γDHw) (10) (10) hm 2 Accelerated gradient projection algorithm Algorithm 2 Accelerated gradient projection algorithm v γ η βt = t/(t +3) Tsub Input: Observation , parameter , step size , , and subiteration number . w(0) = 0 z(0) = w(0) 1: , t = 1,2,...,Tsub 2: for do w(t) = PS ( z(t−1) −η∇zG(z(t−1)) ) 3: z(t) = w(t) +βt(w(t) −w(t−1)) 4: 5: end for x = PC(v−γDHw(Tsub)) 6: v γ η βt = t/(t +3) Tsub Input: Observation , parameter , step size , , and subiteration number . w(0) = 0 z(0) = w(0) 1: , t = 1,2,...,Tsub 2: for do w(t) = PS ( z(t−1) −η∇zG(z(t−1)) ) 3: z(t) = w(t) +βt(w(t) −w(t−1)) 4: 5: end for x = PC(v−γDHw(Tsub)) 6: t = 1,2,...,Tsub 2: for do w(t) = PS ( z(t−1) −η∇zG(z(t−1)) ) 3: z(t) = w(t) +βt(w(t) −w(t−1)) 4: 5: end for Algorithm 1 Accelerated proximal gradient algorithm The proximal gradient framework 3 by minimizing the non-differentiable via its proximal operator , which is defined as 128 ∥w∥∞≤λ (8) (8) ∥w∥∞= maxi \|wi\| x⋆ w⋆ where . As shown in the Supplementary Document, the desired primal optimal solution is related to the dual optimal solution through the following equation: ∥w∥∞= maxi \|wi\| x⋆ w⋆ where . As shown in the Supplementary Document, the desired primal optimal solution is related to the dual optimal solution through the following equation: x⋆= PC(v−γDHw⋆) (9) (9) proxγR(v) = argmin x R(x)+ 1 2γ∥x−v∥2 2 (6) (6) PS (·) S G(w) The problem of Eq. 7 can be effectively solved using the gradient projection algorithm, where the dual variable is iteratively updated by a gradient step followed by a projection operation onto the set . An accelerated variant of the algorithm is presented in Algorithm 2. The Wirtinger gradient of the dual objective function is given by 131 γ > 0 R(x) where is the step size. Notice that Eq. 6 coincides with an image denoising problem, with serving as a regularization function. For the CCTV regularizer, the denoising subproblem can be solved efficiently using the gradient projection algorithm introduced in the next subsection. The proximal gradient algorithm proceeds by iteratively applying a gradient update and proximal update. A variant using the Nesterov’s acceleration technique was adopted 129, as shown in Algorithm 1. The accelerated proximal gradient (APG) algorithm is also known as the fast iterative shrinkage thresholding algorithm (FISTA) 130. APG is well known for its superior convergence speed over the basic proximal gradient algorithm in the convex setting. Through numerical studies, we empirically demonstrate that it also helps to speed up convergence for the nonconvex phase retrieval problem. γ > 0 R(x) where is the step size. Notice that Eq. 6 coincides with an image denoising problem, with serving as a regularization function. For the CCTV regularizer, the denoising subproblem can be solved efficiently using the gradient projection algorithm introduced in the next subsection. The proximal gradient algorithm proceeds by iteratively applying a gradient update and proximal update. A variant using the Nesterov’s acceleration technique was adopted 129, as shown in Algorithm 1. The accelerated proximal gradient (APG) algorithm is also known as the fast iterative shrinkage thresholding algorithm (FISTA) 130. APG is well known for its superior convergence speed over the basic proximal gradient algorithm in the convex setting. Hyperparameter selection Tsub Tsub Tsub λ = 1×10−3 3×10−3 1×10−2 3×10−2 Tsub Tsub = 10 Tsub Tsub 1% Tsub λ λ λ We first investigated the influence of the subiteration number of the gradient projection denoising algorithm on the convergence of the proximal gradient algorithm. As previously pointed out, is a key hyperparameter that should be determined beforehand. We ran the APG algorithm for 1000 iterations using different values ranging from one to ten. Meanwhile, four regularization parameters, , , , and were considered for comparison. To quantify the convergence behavior of the inexact APG algorithms, we calculated the relative error of the final objective function value for each relative to that with . Fig. 6a indicates that the relative error decreases monotonically as increases, and thus, a more accurate proximal update can be obtained by increasing the number of subiterations. We empirically selected the smallest that yielded an error below as the optimal choice. Another observation made from Fig. 6a is the dependence of on regularization parameter . As increases, one needs more subiterations for the denoising algorithm to obtain an adequate solution. This seems quite natural, considering the nonlinear nature of the denoising operation. A larger imposes a stronger regularization force on the image, thus requiring more linear iterative steps to approximate the solution. C η Theorem 2 Assuming that the constraint set is closed and convex, the accelerated gradient projection algorithm for the problem of Eq. 7 converges to the global optimum using a fixed step size that satisfies η ≤ 1 γ2ρ(DH D) (12) (12) γ D ρ(DH D) ≤8 γ = 2 η = 1/32 Theorem 2 suggests that the step size for the denoising algorithm can also be determined beforehand based on given by Theorem 1 and the sparsifying transform . Specifically, for the finite-difference operator, we show in the Supplementary Document that , which, together with , suggests for this particular case. The convergence established by Theorem 1 assumes that an exact solution to the denoising subproblem is obtained at each iteration. The algorithm may converge to a suboptimal solution when the proximal update is inexact, as with the CCTV regularizer. Readers can refer to Ref. 133 and references therein for convergence results regarding inexact proximal gradient algorithms. Therefore, the iteration number used for the gradient projection algorithm should be chosen as a balance between reconstruction quality and computation efficiency. ρ(·) where denotes the spectral radius. The above theorem is an extension of the gradient descent algorithm 132 to the more general class of the proximal gradient algorithms. A Q C Q = F−1diag(h)F F h \|h\| = 1 Q QHQ = I I ρ(AH A) ≤1 γ = 2 The sampling matrix depends only on the physical model. Thus, the step size can be known a priori. Recall that the sampling matrix comprises a circular convolution operator and an image cropping operator . Circular convolution can be efficiently calculated via FFTs as , where is the Fourier matrix and is the transfer function. Note that . Thus, is a unitary matrix that satisfies , where is the identity matrix. It can be easily verified that , which, according to Theorem 1, implies that we should use step size for the proximal gradient algorithm. This choice of step size is in fact nontrivial and generally applicable to any passive measurement scheme. x(0) = AHy In all numerical studies, the algorithms were initialized with an initial estimate of . The algorithms were implemented using MATLAB on a personal computer with Core i7-11700F CPU @ 2.50GHz (Intel) and 16 GB of RAM. The MATLAB code is publicly available in Ref. 134. The next theorem establishes the convergence of the accelerated gradient projection algorithm to solve the convex denoising subproblem via its dual (Eq. 7). The proof can be found in the Supplementary Document. Hyperparameter selection λ λ Another hyperparameter of great importance is the regularization parameter , which offers a tradeoff between model consistency and signal priors. We compared different selections of in terms of both the reconstruction quality and computational efficiency. Fig. 6b plots the CCTV denoising algorithm C The proximal update step in Eq. 6 involves solving a CCTV-regularized image denoising subproblem. It is a convex optimization problem when is closed and convex. No exact closed-form solution to this problem is available. Thus, an iterative solver should be invoked at every iteration. We followed the approach proposed in Refs. 95,131 and consider solving the primal problem via its βt ≡0 Theorem 1 The basic proximal gradient algorithm (with in Algorithm 1) for the problem of Eq. 3 converges to Page 10 of 17 Gao et al. Light: Advanced Manufacturing (2023)4:6 γ a stationary point using a fixed step size that satisfies [0,π] experiment section to quantitatively evaluate the algorithmic behaviors and further shed light on our empirical observations and theoretical results. The wavelength was set to 500 nm, the pixel size was 5 μm, and the imaging distance was 5 mm. Eight standard test images of size 256×256 were used to generate the virtual phase objects for the simulation. The phase of the object was assumed to have a range of . The objects have a uniform amplitude distribution and were zero-padded to avoid boundary artifacts. Additive white Gaussian noise was added to each hologram such that the signal-to-noise ratio (SNR) was 20 dB by default. We calculated the phase RMSE as a metric to quantitatively compare the reconstruction quality of different methods. γ ≤ 2 ρ(AH A) (11) (11) Discussion a 0 1 2 3 4 5 6 7 8 9 10 0 4 8 12 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 0 3 6 0 1 2 3 Subiteration number Subiteration number Subiteration number Subiteration number Relative error ×10−3 λ = 1 × 10−3 ×10−2 λ = 3 × 10−3 ×10−2 λ = 1 × 10−2 ×10−1 λ = 3 × 10−2 a 0 10 20 30 40 50 0.7 0.6 0.5 0.4 0.3 0.2 b c RMSE (rad) Runtime (s) λ = 1 × 10−3 λ = 3 × 10−3 λ = 1 × 10−2 λ = 3 × 10−2 c λ = 1 × 10−2 0.1449 rad λ = 3 × 10−2 0.1793 rad λ = 1 × 10−3 0.2834 rad λ = 3 × 10−2 0.1793 rad λ = 3 × 10−3 0.1698 rad λ = 1 × 10−2 0.1449 rad Tsub λ λ Tsub Fig. 6 Comparison of different hyperparameter selections based on simulations. a Relative error against subiteration number for different regularization parameters . The dashed lines indicate the value of 1%, and the downward-pointing triangles indicate the optimal choice of subiteration number that satisfies the error criterion. b Root-mean-square error against runtime for different when using the corresponding optimal in a. c Phase obtained by running the algorithms for 50 s. The second row shows the enlarged areas. The regularization parameters and RMSEs are shown below. No physical constraints are considered in this simulation. λ to 30 dB to the simulated hologram, as shown in Fig. 8a. The corresponding phases retrieved by the CCTV-AS method are presented in Fig. 8c. The CCTV method can achieve high-quality phase retrieval under moderate-noise conditions. However, when the hologram is severely degraded by measurement noise, a larger regularization parameter is required to suppress reconstruction artifacts. The retrieved phase may then exhibit a staircase artifact when the regularization parameter is too large, which is an inherent problem for TV-based regularizers. Therefore, more advanced image priors are required to achieve better reconstruction quality under noisy conditions. Discussion We performed numerical studies based on the same in- line holographic imaging model as described in the Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 11 of 17 a 0 1 2 3 4 5 6 7 8 9 10 0 4 8 12 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 4 5 6 7 8 9 10 0 1 2 3 0 3 6 0 1 2 3 Subiteration number Subiteration number Subiteration number Subiteration number 0 10 20 30 40 50 0.7 0.6 0.5 0.4 0.3 0.2 b c (rad) 0 π Relative error ×10−3 λ = 1 × 10−3 λ = 1 × 10−3 0.2834 rad ×10−2 λ = 3 × 10−3 λ = 3 × 10−3 0.1698 rad ×10−2 λ = 1 × 10−2 ×10−1 λ = 3 × 10−2 RMSE (rad) Runtime (s) λ = 1 × 10−2 0.1449 rad λ = 3 × 10−2 0.1793 rad λ = 1 × 10−3 λ = 3 × 10−3 λ = 1 × 10−2 λ = 3 × 10−2 Tsub λ λ Tsub Fig. 6 Comparison of different hyperparameter selections based on simulations. a Relative error against subiteration number for different regularization parameters . The dashed lines indicate the value of 1%, and the downward-pointing triangles indicate the optimal choice of subiteration number that satisfies the error criterion. b Root-mean-square error against runtime for different when using the corresponding optimal in a. c Phase obtained by running the algorithms for 50 s. The second row shows the enlarged areas. The regularization parameters and RMSEs are shown below. No physical constraints are considered in this simulation. Discussion Tsub λ Tsub = 1 λ = 1×10−3 Tsub = 2 λ = 3×10−3 Tsub = 5 λ = 1×10−2 Tsub = 8 λ = 3×10−2 λ = 1×10−2 convergence curves against runtime, where the optimal was used for each , that is, for , for , for , and for . A larger regularization parameter can help accelerate the convergence at the beginning stage. However, the final result tends to be over-smoothed, as shown in Fig. 6c. Therefore, we set for balanced performance in terms of speed and accuracy. Generalization to different scenes To further examine the generalizability of the CCTV model, we tested the algorithm on various test images and compared it with IP and CTV methods. The results are shown in Fig. 9. The conventional IP algorithm generally suffers from a high reconstruction error because the physical constraints alone are insufficient to suppress the twin-image artifact. On the other hand, the CTV model often yields visually appealing results, yet its performance is highly dependent on the sparsity of the scene. For example, the CTV model tends to perform well for scenes with very sparse gradients, such as the House image, but it may be unsuitable for retrieving complex scenes, such as the Goldhill and Mandrill images. In all cases, the CCTV model offers the best reconstruction quality owing to the Comparison of physical constraints 256×256 Having determined the optimal hyperparameters for the complex TV, we evaluated the effect of incorporating additional physical constraints. Fig. 7 shows the results obtained using no constraints, support constraint, absorption constraint, or both. For the support constraint, the support region was set to the central pixels of the image. The introduction of the absorption constraint can significantly improve the overall quality, while the support constraint can help suppress artifacts at the boundaries. Incorporating physical constraints also helps accelerate convergence. Influence of measurement noise The inherent ill-posedness of single-shot phase retrieval leads to high sensitivity to measurement noise. To quantitatively compare the phase reconstruction quality under noisy conditions, we added different levels of additive white Gaussian noise with an SNR ranging from 5 Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 12 of 17 0.6 0.5 0.4 0.3 0 50 100 150 200 Iteration RMSE (rad) (rad) (rad) 1 −1 0 π a b CTV 0.3099 rad CTV CCTV-S CCTV-A CCTV-AS CCTV-S 0.2866 rad CCTV-A 0.2496 rad CCTV-AS 0.2111 rad Fig. 7 Comparison of convergence speed and reconstruction quality with different physical constraints. a The RMSE is plotted against the iteration number. b The upper and lower rows show the reconstructed phase after 200 iterations and the corresponding phase residuals, respectively. The calculated phase RMSEs are shown below. 0.6 0.5 0.4 0.3 0 50 100 150 200 Iteration RMSE (rad) a b CTV CCTV-S CCTV-A CCTV-AS b Fig. 7 Comparison of convergence speed and reconstruction quality with different physical constraints. a The RMSE is plotted against the iteration number. b The upper and lower rows show the reconstructed phase after 200 iterations and the corresponding phase residuals, respectively. The calculated phase RMSEs are shown below. SNR = 30 dB SNR = 20 dB SNR = 10 dB SNR = 5 dB 0 6 0.8 −0.8 (rad) (rad) 0 π a b c d 0.0699 rad 0.1045 rad 0.1994 rad 0.2805 rad Fig. 8 Effect of measurement noise on reconstruction quality. a Simulated noisy holograms with different SNRs. b Corresponding enlarged areas of a. c Retrieved phase by the CCTV-AS method. d Phase residuals of c. The calculated phase RMSEs are shown below. b b c d 0.1994 rad 0.0699 rad 0.1045 rad 0.1994 rad 0.2805 rad Fig. 8 Effect of measurement noise on reconstruction quality. a Simulated noisy holograms with different SNRs. b of a. c Retrieved phase by the CCTV-AS method. d Phase residuals of c. The calculated phase RMSEs are shown be λ = 1×10−2 λ combination of sparsity priors and physical constraints. We also set as fixed throughout the simulation experiments. This implies that the selection of is not very sensitive to the scene content. problem by comparing the APG algorithm with the non- accelerated version in both unconstrained and constrained cases, as shown in Fig. 10a, b, respectively. Nesterov’s method significantly improved the convergence speed in both cases. Influence of measurement noise The APG algorithm involves only an additional extrapolation step per iteration, whose time consumption is negligible compared with the gradient and proximal update steps. The empirical success of Nesterov’s method can be Nesterov acceleration for nonconvex optimization We also empirically validated the effectiveness of Nesterov’s method for the nonconvex phase retrieval Gao et al. Light: Advanced Manufacturing (2023)4:6 Page 13 of 17 0.6 0 100 150 Iteration IP-AS CTV CCTV-AS 50 0 100 150 Iteration 50 0 100 150 Iteration 50 0 100 150 Iteration 50 0 100 150 Iteration Iteration 50 0.6 0.6 0.8 0.7 0.5 0.3 0.4 0.3 0.2 0.3 0.2 0 100 150 50 200 (rad) 0 π RMSE = 0.4774 rad RMSE = 0.5304 rad RMSE = 0.5679 rad RMSE = 0.7365 rad RMSE = 0.6215 rad RMSE = 0.3887 rad RMSE = 0.3202 rad RMSE = 0.4692 rad RMSE = 0.2765 rad RMSE = 0.3589 rad RMSE = 0.2973 rad RMSE = 0.3003 rad RMSE = 0.2311 rad RMSE = 0.3383 rad RMSE = 0.2188 rad RMSE = 0.1961 rad RMSE = 0.2063 rad RMSE = 0.1767 rad IP-AS CTV CCTV-AS RMSE (rad) Fig. 9 Comparison of the IP-AS, CTV, and CCTV-AS methods for various test images (from left to right: Ship, Goldhill, House, Lake, Male, Mandrill). The upper three rows show the reconstructed phase by the three methods. The bottom row plots the RMSE against the iteration number. IP-AS Fig. 9 Comparison of the IP-AS, CTV, and CCTV-AS methods for various test images (from left to right: Ship, Goldhill, House, Lake, Male, Mandrill). The upper three rows show the reconstructed phase by the three methods. The bottom row plots the RMSE against the iteration number. sparse nature of complex real-world scenes while leveraging physically tractable constraints to further improve imaging quality, as confirmed by both simulated and optical experiments. In addition, we conducted a detailed theoretical analysis from a mathematical viewpoint, which provides general guidance for algorithm parameter selection to ensure convergence, obviating the need for laborious manual tuning. We emphasize that the proposed framework is generally applicable to various optical configurations and can easily adapt to other physical constraints and sparsifying operators. 0 100 200 200 300 400 180 Objective value 200 220 240 Iteration 0 100 200 Iteration a Objective value b PG APG PG APG PG APG Fig. 10 Convergence of the proximal gradient (PG) algorithm and its accelerated variant (APG) in a unconstrained case and b constrained case. 180 Objective value 200 220 240 0 100 200 Iteration a PG APG 0 100 200 200 300 400 Iteration Objective value b PG APG PG APG b Fig. Nesterov acceleration for nonconvex optimization 10 Convergence of the proximal gradient (PG) algorithm and its accelerated variant (APG) in a unconstrained case and b constrained case. Although numerically extending the proposed algorithmic framework to coherent diffraction imaging is straightforward, retrieving complex-valued images from a single far-field diffraction pattern remains challenging. 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https://openalex.org/W4200051469	https://revistas.ucm.es/index.php/GERI/article/download/78127/4564456559126	es		Estilo de vida durante el priscilianismo: aproximaciones bioarqueológicas	Gerión	2,021	cc-by	15,017	ARTÍCULOS Gerión. Revista de Historia Antigua ISSN: 0213-0181 https://dx.doi.org/10.5209/geri.78127 Estilo de vida durante el priscilianismo: aproximaciones bioarqueológicas1 Olalla López-Costas2 Recibido: 15 de febrero de 2021 / Aceptado: 21 de mayo de 2021 Resumen. Trato de reconstruir aquí las principales características de la vida diaria durante el priscilianismo en Galicia. A través de una revisión bibliográfica de las aproximaciones bioarqueológicas, que van desde evidencias aportadas por las turberas, los restos vegetales y animales, a los esqueletos humanos, reconstruyo las principales características del siglo IV al VI y sus diferencias con el periodo anterior. Los puntos más destacados son un súbito empeoramiento del clima, una mayor intensidad de la erosión de los suelos y de la destrucción de los bosques, además de la probable modificación de los hábitos alimentarios, incluyendo una mayor dependencia de productos marinos y mijos. En base a esta información, intento comprender si alguna de estas modificaciones pudo responder o estar conectada con esta nueva forma de vida religiosa en el noroeste peninsular o, al menos, comprender mejor cómo era la vida diaria de las personas. Palabras clave: paleodieta; paleoambiente; Antigüedad tardía; Galicia; clima; contaminación. [en] Lifestyle during Priscillianism: Bioarchaeological Approaches Abstract. By analyzing bioarchaeological proxies –from evidence provided by peat records, plant and animal remains, to human skeletons– I aim to reconstruct the main features sketching lifestyle during Priscillianism (AD 4th to 6th centuries) as compared to the previous period by reviewing published literature. A sudden climate deterioration, increase in soil erosion, forest decline and a possible modification in diet, which became more dependent on marine resources and millets, are the main observed findings. Based on this information, I try to understand if some of these characteristics responded to, directly or indirectly, the new religious way of life in Nonwestern Iberia, or at least to understand better the everyday life of these societies. Keywords: Paleodiet; Paleoenvironment; Late Antiquity; Galicia; Climate; Pollution. Sumario. 1. Reconstruyendo el estilo de vida. 2. Un clima cambiante. 3. El no retorno del bosque y el cambio en el uso del suelo. 4. Modificaciones en la explotación de los metales. 5. Una alimentación que se adapta. 6. Breves apuntes sobre la morfología, salud, enfermedad y costumbres funerarias. 7. La vida en tiempos de Prisciliano y de los priscilianistas, una reflexión. 8. Referencias bibliográficas. Cómo citar: López-Costas, O. (2021): Estilo de vida durante el priscilianismo: aproximaciones bioarqueológicas, en Gerión 39(2), 775-800. 1 2 Este trabajo fue redactado bajo la financiación del proyecto “Pollutio: Contaminación metálica ambiental en poblaciones de Hispania entre los siglos I al V a través de los restos óseos” - Plan Nacional Retos JIN 2019 (PID2019-111683RJ-I00) Ministerio de Ciencia e Innovación. Proyecto realizado con la Beca Leonardo a Investigadores y Creadores Culturales 2020 de la Fundación BBVA. Mis agradecimientos a Antonio Martínez Cortizas y a Pedro López Barja por sus comentarios y ayuda. Universidade de Santiago de Compostela – Stockholm University. E-mail: olalla.lopez@usc.es ORCID: 0000-0002-6499-226X Gerión, 39(2) 2021: 775-800 775 López-Costas, O. Gerión, 39(2) 2021: 775-800 776 1. Reconstruyendo el estilo de vida3 El estilo de vida es uno de los aspectos que nos acercan en mayor medida a lo que supone el día a día de una sociedad. Existen varias definiciones y muchas maneras de entenderlo, que van desde lo individual a lo colectivo.4 Lo concibo como las particularidades que definen la vida diaria desde un punto de vista directo y físico; tales como qué salud tienes, cuál es tu dieta, a qué actividades dedicas tus días o tus posibilidades de obtener los recursos necesarios. Estas se unen a otras externas o más amplias que terminan de perfilar la identidad individual, tales como la clase social, las creencias, la cultura, la economía y finalmente el ambiente físico. Pertenecen las primeras a aquellas preguntas que se pueden intentar responder desde el estudio de los restos humanos (osteología, osteoarqueología, antropología física). Otras disciplinas como la zooarqueología o la paleobotánica nos ayudan a distinguir el uso que una comunidad hace de los recursos animales y vegetales y, aunque no proporcionan información directa de las personas, son buenos proxies (marcadores, aproximaciones). En lo relativo a las segundas, se realiza un salto a una mayor escala y es aquí donde las reconstrucciones paleoambientales, la arqueología funeraria o la arqueotanatología juegan su papel. Los aspectos y disciplinas mencionadas se relacionan entre sí englobadas en un supra-concepto que podríamos denominar bioarqueología o uso de los proxies biológicos para reconstruir el pasado de los seres humanos. En todos los niveles caracterizados previamente, la historiografía proporciona información legítima. El complemento de la bioarqueología surge de la existencia de periodos o atributos “oscuros” no fácilmente abarcables por falta de fuentes; en otros casos son algunos grupos humanos los que están silenciados (ej. mujeres, enfermos, niños), o simplemente puede proporcionar consideraciones adicionales. Complementarias una de la otra, la historiografía y la bioarqueología tienen, en mi opinión, más futuro si siguen un camino común, rehuyendo los recorridos paralelos que les han caracterizado hasta ahora. Como parte de este número especial sobre Prisciliano, en estas líneas trato de reunir todos los aspectos en los que la bioarqueología proporciona información (no solo datos) sobre un periodo concreto, aquel donde el priscilianismo estuvo vigente (siglos IV al VI), y su inmediato anterior y posterior,5 mediante una revisión crítica de los estudios publicados hasta el momento. Trabajo aquí con una gran variedad de “archivos” bioarqueológicos y paleoambientales, que al igual que si se tratara de las fuentes documentales, almacenan la información del pasado en señales macroscópicas, físicas y químicas. El área que acoto es la actualmente constituida por la comunidad autónoma de Galicia, en el noroeste de España. En los siguientes apartados pongo especial énfasis en señalar las características que particularizan cada tipo de proxy y archivo bioarqueológico, así como sus certidumbres y sus limitaciones, para que los lectores puedan desarrollar una idea de conjunto de la envergadura de cada uno dentro de mi discurso. Mi objetivo final es aportar información sobre la vida cotidiana de las gentes que vivieron en la Antigüedad Tardía, siendo conscientes 3 4 5 Todas las fechas de este artículo son d.C. a menos que se especifique lo contrario. Sobel 2013, 7-30, lo define como “modo de vida” y concluye que no está claro si el estilo de vida pertenece a un individuo o a agregaciones de individuos. Me alejo de las definiciones filosóficas o relacionadas con el estatus socioeconómico debatidas por este autor, y me centro en un aspecto físico. Ver los trabajos de Chadwick 1976; Escribano Paño 1996; Piay 2019. López-Costas, O. Gerión, 39(2) 2021: 775-800 777 de que desgraciadamente los “archivos” analizados nunca nos podrán informar de cómo pensaban o cómo sentían.6 2. Un clima cambiante El final del periodo romano constituye un momento de cambios que afectaron a los seres humanos no solo cultural o socialmente, sino también a nivel climático. En Galicia tenemos la ventaja de contar con una considerable extensión de turberas en algunas zonas de montaña, como la sierra del Xistral. Los depósitos de turba acumulan a lo largo del tiempo señales de cambios ambientales dejando un registro, en principio imperturbable, del ambiente de ese momento. Aspectos tales como las condiciones climáticas o el registro metálico, proxy de contaminación ambiental, quedan registrados en la turba que se forma coetáneamente (mediante su incorporación a través de los organismos vivos (plantas) o su deposición directa desde la atmósfera) y son progresivamente englobados en el depósito turboso a medida que este aumenta en espesor por formación de nuevas capas de turba en superficie. El depósito acumulativo de la misma hace que las modificaciones en profundidad sean limitadas, por lo que dichas características del ambiente quedan archivadas en la materia orgánica e inorgánica que conocemos como turba, cuya edad aumenta con la profundidad. En estas turberas se han obtenido registros que retroceden en el tiempo hasta 10.000 años, proporcionando una visión amplia de los cambios en el ambiente durante periodos de mayor o menor presión humana sobre el mismo. Por esa razón se las conoce como “archivos ambientales”, ya que funcionan de manera similar a las fuentes escritas, encapsulando la información de un determinado momento. Los principales archivos ambientales para reconstruir el clima son los hielos polares, los sedimentos finos de lagos y las turberas. De estos archivos, para Galicia, y debido a su extensión, las últimas permiten hacer una reconstrucción regional. En el caso que nos atañe, ha sido posible obtener una reconstrucción climática de los últimos 4.000 años a partir de la turbera de Penido Vello (Serra de O Xistral, norte de Lugo), la cual revela importantes cambios climáticos en el periodo objeto de este trabajo.7 Desde el cambio de era se puede observar un aumento de las temperaturas constituyendo el periodo conocido como Óptimo Climático Romano y que se caracteriza en esta zona por temperaturas elevadas con alguna caída menor y una humedad estable y relativamente baja.8 A partir del siglo IV se observa una modificación en la humedad, la cual se vuelve más inestable, con aumentos significativos pero cambiantes entre el 300 y el 600.9 Sin embargo, no hay evidencias de cambios en los gradientes de humedad con respecto a la actualidad,10 es decir, las zonas de Galicia que son relativamente más secas ahora también lo habrían sido en el pasado, y lo mismo sucede con las más húmedas. Un intenso aumento de la 6 7 8 9 10 La información de carácter histórico-social no basada en estudios bioarqueológicos es abordada de manera más sucinta por razones de espacio y para evitar redundancias con otros trabajos de este número especial. Martinez Cortizas et alii 1999. Mighall et alii 2006. Mighall et alii 2006. Martínez Cortizas – Pérez Alberti 1999. 778 López-Costas, O. Gerión, 39(2) 2021: 775-800 humedad se detecta en torno al siglo IV.11 A esta evolución variable se le une un deterioro térmico relativamente súbito desde mediados del siglo IV, que alcanza su mínimo de temperaturas en torno al 500. Las temperaturas se empezaron a recuperar ligeramente alrededor de un siglo después.12 Uno de nuestros estudios recientes ha condensado los índices de humedad en uno solo, obteniendo para el noroeste una cronología similar a la anteriormente explicada.13 Para la comprensión de esta información y su grado de validez en el trabajo de reconstrucción histórica, ha de tenerse en cuenta que la información proporcionada por las turberas, aunque está directamente conectada al ámbito local, a nivel climático refleja modificaciones a escala regional. Por lo tanto, se trata de una buena aproximación al clima en el noroeste de España, o al menos la mejor aproximación que tenemos. No obstante, dicha reconstrucción cuenta con desventajas propias del archivo que se estudia. Los testigos de turba suelen tener una resolución muy por debajo de la preferencia del historiador, puesto que los análisis se realizan en secciones de 1 a 5 cm, que representan periodos relativamente largos de tiempo (de décadas a centurias). Asimismo, es importante considerar los modelos cronológicos construidos en estos trabajos, es decir, asegurarnos de que cuentan con suficientes dataciones y son robustos. Finalmente, en nuestra interpretación de estas variaciones, como historiadores hemos de tener presente que por muy bueno que sea dicho modelo siempre posee incertidumbres, de modo que ligeras modificaciones en el modelo cronológico pueden llevarnos a cambios en la asignación temporal. Por ello, el deterioro climático que se observa claramente a inicios del siglo IV pudo, en realidad, haber ocurrido unas décadas antes o después. 3. El no retorno del bosque y el cambio en el uso del suelo Ligado o no a este deterioro climático, las turberas y otros archivos ambientales, como los suelos coluviales, han revelado modificaciones importantes en la extensión de los bosques. El periodo romano se caracterizó por una caída en la cobertura arbórea, pero también por un aumento de especies cultivadas o manejadas, como el castaño o el olivo14 (reflejado en el aumento del polen de dichas especies en los registros polínicos). Sin embargo, el estudio de turberas ha revelado una recuperación de la cobertura arbórea asociada al periodo que nos atañe y que empieza en torno a mediados del siglo IV, alcanzando el máximo hacia el 450, con ligeras variaciones según los testigos analizados.15 Esto indica que hubo un descenso en la presión sobre las principales especies de árboles que forman el bosque atlántico, es decir, en los registros aumenta, entre otros, el polen de las especies del género Quercus, lo que se interpreta como una recuperación de la superficie ocupada por robledales. Las variaciones cronológicas con respecto al máximo de esta recuperación son fruto de la anteriormente mencionada 11 12 13 14 15 Schellekens et alii 2011, en muestras datadas entre los años 270-385. Se observa también un máximo de humedad en una muestra datada en 640, no incluida al tratarse de una medida puntual. Martínez Cortizas et alii 1999. Martínez Cortizas et alii 2019. Martínez Cortizas et alii 2005; Silva-Sánchez et alii 2014. Muñoz Sobrino et alii 1997; Martínez Cortizas et alii 2005; Mighall et alii 2006; Silva-Sánchez et alii 2014; Martínez Cortizas et alii 2019. López-Costas, O. Gerión, 39(2) 2021: 775-800 779 incertidumbre en el modelo cronológico y la variabilidad cronológica local, ya que se cubre una geografía amplia (archivos situados en el Xistral hasta los Ancares y los Montes de O Bocelo). Los estudios polínicos también indican que a este periodo de breve recuperación sigue una drástica reducción del bosque. Esta bajada empieza en torno al 500 y alcanza un mínimo de cobertura arbórea entre el 600-700.16 Si bien existen variaciones que llegan a situar esta caída en fechas tan tardías como el siglo X,17 se trata de registros con una resolución cronológica baja (pocas dataciones radiocarbónicas) por lo que empleamos preferentemente aquellos que cuentan con modelos cronológicos más fiables. En algunas zonas de montaña del interior, como los Ancares y O Courel (este y sur de Lugo, respectivamente), se observa un aumento del abedul en substitución del robledal,18 en la sierra do O Bocelo, cerca de Santiago de Compostela, aumentan los alisos, probablemente ligados a la mayor humedad,19 mientras que en el Xistral se produce un descenso de todas las principales especies arbóreas.20 Esto indica que existió una diversidad de respuestas. Esta deforestación parece afectar a toda la fachada norte, ya que se observan respuestas similares en Asturias y Cantabria.21 No obstante, si algo caracteriza a este periodo es el aumento del brezal o monte bajo. Paralelo a la disminución del polen de roble y otros árboles que constituían el monte gallego, se detecta una mayor proliferación de especies de matorral: sobre todo de ericáceas (como la uz o Erica arborea L.), retamas y aulagas (tojos).22 Estas son especies clave del brezal atlántico y oromediterráneo, que aparentemente no tuvieron en este momento un gran uso por las poblaciones, ya que los estudios antracológicos sugieren que se seguía prefiriendo la madera de árboles, como los robles o fresnos, para calentarse.23 Lo que sí probablemente sucedió fue un cambio en el paisaje del monte gallego, el cual ahora se vuelve más bajo y “pelado”. Este clareo probablemente modificó la percepción del espacio, siendo posible abarcar mayores extensiones con la vista. La transición entre el robledal y la combinación brezal-abedul no se puede interpretar como el resultado de un único factor, ya sea este natural o humano. Más bien, fue la consecuencia de una sinergia entre el deterioro climático y un aumento en la presión por parte de las gentes. Estudios en otro tipo de archivos ambientales, los suelos coluviales, revelan un nuevo factor ligado tradicionalmente en el noroeste peninsular al manejo del bosque, el uso del fuego.24 En las sierras del interior de Galicia, un trabajo con baja resolución para el periodo que nos interesa ha revelado un aumento en la intensidad de los fuegos en algún momento a partir del cambio de era.25 En suelos de Campo Lameiro (Pontevedra), una zona de media elevación, se observa una intensificación del régimen de incendios en torno al 500 (registrado como un 16 17 18 19 20 21 22 23 24 25 Muñoz Sobrino et alii 1997; Martínez Cortizas et alii 2005; Mighall et alii 2006; Silva-Sánchez et alii 2014. Muñoz Sobrino et alii 2005. Muñoz Sobrino et alii 1997; Santos et alii 2000. Silva-Sánchez et alii 2014, los cambios de humedad se refieren a los índices reconstruidos en el propio testigo y variaciones regionales. Mighall et alii 2006. Para Asturias ver López-Merino et alii 2014; para Cantabria Jalut et alii 2010. Muñoz Sobrino et alii 1997; Martínez Cortizas et alii 2005; Mighall et alii 2006. Martín Seijo 2013. Kaal et alii 2011. Santos et alii 2000. 780 López-Costas, O. Gerión, 39(2) 2021: 775-800 aumento en el contenido en carbones)26 paralelo a una mayor presencia de ericáceas, jaras, cereales y algunas herbáceas,27 y al aumento de la erosión.28 Este hecho, unido a la presencia de llantén menor (Plantago lanceolata L.), especie típica de praderas, apunta según estos autores a un posible clareo del bosque para crear pastos.29 En las investigaciones realizadas en turberas también se ha detectado un aumento del polen de cereal y de esporas de hongos coprófilos que indican actividades agrícolas y ganaderas.30 Ambos hechos se unen a una acentuación en la erosión del suelo y una mayor presencia de carbones,31 así como de todas las especies de herbáceas,32 lo cual apunta en la misma dirección de intensificación de la ganadería y posible aumento de la agricultura después de la recuperación de los siglos IV y V. El estudio de suelos aterrazados llevado a cabo en las inmediaciones de Santiago de Compostela muestra la misma intensificación del uso del suelo entre el final del periodo romano y el siglo VI (hay un hiato en la cronología que no permite ser más exactos).33 El aclarado muy posiblemente tuvo lugar mediante quemas controladas previas al aterrazamiento. En esta última zona también se documenta el uso de matorral bajo, como las aulagas (tojos), para la fertilización. El aterrazado intenso de Galicia parece haber tenido un origen en este periodo, intensificándose a partir del siglo V.34 Como nota, ha de recordarse que los suelos coluviales son archivos de baja resolución cronológica si se los compara con las turberas, por lo que, cuando se trabaja con estudios realizados en ellos, han de tomarse las fechas con más cautela.35 Además de la necesidad de pastos para la ganadería y nuevas tierras para la agricultura, el aumento de la presión humana sobre el bosque pudo estar condicionado también por el clima.36 El observado empeoramiento de las condiciones climáticas, con inviernos más fríos, pudo incrementar la necesidad de “productos” del bosque en forma de leña. Estudios antracológicos de yacimientos arqueológicos romanos muestran un mayor consumo de madera de roble y de fresno para calentarse o cocinar.37 En Cova do Xato, en la sierra de O Courel, cueva que parece haber estado ocupada durante los periodos de inestabilidad climática de los siglos IVV, se encontró una mezcla de maderas de combustión lenta, roble y retama, junto con otras de sauce o chopos para avivar el fuego.38 Por otro lado, cambios en las necesidades constructivas también influyen en la tala de árboles; el roble, el fresno 26 27 28 29 30 31 32 33 34 35 36 37 38 Kaal et alii 2008. Carrión et alii 2010, 59, fig. 5 (diagrama), y 62; en el diagrama se aprecia el aumento de Erica sp, Erica arborea L. y Cistus. Martínez Cortizas et alii 2009, 198, fig. 1, donde se puede ver la coincidencia de los momentos de mayor humedad obtenidos de Mighall et alii 2006 con el mayor contenido de carbones y erosión. El valor de pH disminuye volviéndose el suelo más ácido, alcanzando el valor mínimo de pH y por lo tanto el máximo de acidez, lo cual pudo cambiar la fertilidad de los suelos haciéndose necesario el uso de fertilizantes vegetales o del aterrazamiento. Kaal et alii 2008; Kaal et alii 2011. Silva-Sánchez et alii 2014. Martínez Cortizas et alii 2005. Mighall et alii 2006. Ferro Vázquez et alii 2014; Ferro Vázquez et alii 2015; ver discusión sobre el uso de los suelos en la zona de Gaiás de Santiago de Compostela. Ballesteros-Arias 2010, analiza 23 elementos del espacio agrario situados mayormente en Pontevedra y el norte de Lugo. Variaciones en el entorno de las centurias, sin que se dejen de situarse en la Tardoantigüedad. Los datos de los que dispongo no me permiten saber si es consecuencia de un incremento demográfico o no. Martín Seijo 2013. Teira Brión et alii 2012. López-Costas, O. Gerión, 39(2) 2021: 775-800 781 y otras arbóreas se han encontrado en yacimientos del siglo III al V formando vigas, canalizaciones de aguas y hasta cestos.39 Un travesaño de madera de roble fue encontrado en Gijón (Asturias) como parte de un mecanismo para sacar agua de un pozo (cargadero), así como un cubo del mismo material.40 Otras actividades menos generalizadas en el espacio geográfico, como las salinas, también necesitaban árboles para su mantenimiento, tal y como se observa en Vigo, donde se halló un entretejido de castaño y otras maderas.41 Un último factor que se acopla a los periodos primero de recuperación del bosque y luego de caída de la cobertura arbórea, es la presencia de metales considerados contaminantes. En este caso, la deforestación y la erosión se asocian con las prácticas derivadas de la explotación de los metales (minería y metalurgia),42 aspecto del que hablo en el siguiente apartado. En contra, las prácticas funerarias, y más en concreto la caída en desuso de la cremación, pudieron ayudar a la recuperación del bosque. La inhumación disminuye los requerimientos de madera, que se restringen a las cajas (normalmente de pino), mientras que en las cremaciones del noroeste se requerían grandes cantidades de roble, pino y fresno.43 Como herencia del periodo que estamos tratando, merece la pena reflexionar sobre lo que supuso la pérdida generalizada de los bosques en Galicia a partir del siglo VI. Esta caída fue tan profunda que no se observa retorno en los registros. Es decir, que el bosque nunca se recupera. Así, citando las palabras de Martínez Cortizas: “crea el paisaje gallego que conocemos”, donde predomina el matorral tipo brezal.44 Por tanto, el periodo que nos atañe parece que constituyó el tránsito hacia una nueva realidad del paisaje arbustivo que tanto caracteriza esta zona de la Península Ibérica, siendo el actual una “herencia” fruto de las condiciones climáticas, sociales y culturales sucedidas desde el siglo IV. Para resaltar la importancia de estos cambios del paisaje vegetal del noroeste ibérico, valga mencionar que éstos se condensan en tres tipos de hábitats actualmente protegidos por la red Natura 2000: los brezales húmedos atlánticos de ericáceas, los brezales secos atlánticos costeros de ericáceas y los brezales oromediterráneos endémicos con aulagas (DOG Núm. 62, 31 de marzo de 2014). Los priscilianistas y sus coetáneos condicionaron en gran medida nuestra Galicia actual. 4. Modificaciones en la explotación de los metales En la extracción y procesado de la mayoría de metales, como el oro, la plata, el cobre, el estaño, el mercurio o el plomo, se liberan a la atmósfera partículas (polvo generado en las operaciones mineras) y gases contaminantes (por liberación de vapores durante el procesado de las menas). En el caso del mercurio, además de su minería para usos directos, por ejemplo, en forma de minio como pintura roja, 39 40 41 42 43 44 Martín Seijo – Carrión Marco 2012; Martín Seijo 2013; Costa Vaz et alii 2016. Ver Fernández Ochoa – Gil Sendino – Salido Domínguez 2015; Fernández Ochoa – Salido Domínguez – García Díaz – Gil Sendino 2015; Carrión Marco et alii 2015. Entre los restos de objetos de madera encontrados en el pozo-depósito de la fábrica de tabacos de Gijón muchos estaban realizados con roble y castaño, además de un gran cesto de madera de clemátide. Martín Seijo 2013; Martín Seijo 2019. Martínez Cortizas et alii 2005. Aira Rodríguez – Uzquiano Ollero 1996; Martín Seijo 2013; Martín Seijo – Vila 2019. Ver, por ejemplo, Martínez Cortizas et alii 2005; Martínez Cortizas et alii 2019. López-Costas, O. Gerión, 39(2) 2021: 775-800 782 se usaba en la minería del oro. El mercurio servía para amalgamar el oro (unirse a partículas pequeñas) y posteriormente “refogar” la amalgama, es decir calentarla hasta alcanzar el punto de fusión del mercurio, el cual se volatiliza dejando el oro sólido. En el caso de la plata, esta está presente en minerales como la galena (sulfuro de plomo). Para su purificación se requería un proceso conocido como copelación o fundición en vaso de copela. Dentro de dicha copela se calentaba en ambiente aireado el mineral que contiene la plata causando la oxidación del plomo en forma de litargirio (monóxido de plomo). Este vaso separa la plata del litargirio a la vez que se libera parte de este monóxido en forma de vapores. Otro ejemplo es la producción de bronce, en la cual también se liberan vapores de estaño y cobre. En todos estos ejemplos los elementos metálicos como el plomo, el mercurio o el cobre pasaban a la atmósfera y se dispersaban a escala local, regional o hemisférica, llegando hasta Groenlandia,45 siendo depositados, por tanto, en la superficie terrestre en otro lugar. Al ser considerados metales tóxicos para muchos organismos, su presencia en la atmósfera se incluye dentro de la contaminación atmosférica. De hecho, la minería y la metalurgia fueron las dos principales actividades humanas causantes de contaminación atmosférica en el periodo preindustrial. La atmósfera no es en sí misma un archivo; no obstante, el hecho de que los metales fuesen depositados en diversas regiones del planeta ha permitido su estudio, reconstruyendo la contaminación atmosférica del pasado investigando registros de archivos naturales y bioarqueológicos (turberas, sedimentos de lagos, hielos polares, esqueletos humanos y animales), como veremos a continuación. El periodo romano supuso un cambio radical en la intensidad de la minería y metalurgia. A partir del cambio de era, la alta demanda de productos metálicos por el Imperio trae como consecuencia un aumento de ambas actividades. El estudio de las turberas ha demostrado las consecuencias de ello. Tanto en Galicia como en otras áreas del suroeste de Europa, como los Pirineos, y en el norte de África, se observa un acoplamiento entre la deforestación del bosque, la erosión y el aumento de la contaminación por plomo, lo cual parece tener un origen común: la explotación de los metales.46 La contaminación viaja hasta el norte; en Fenoscandia se registran picos en la concentración de plomo en turberas y lagos. Se trata en estos casos de una contaminación que tiene su origen en el sur de Europa, ya que en Suecia la minería/metalurgia propia no alcanzó estas magnitudes hasta época plenomedieval.47 Este incremento en la contaminación atmosférica por metales es tan marcado en toda Europa y tan definido temporalmente, que el aumento del plomo en el periodo romano ha sido propuesto como un marcador cronológico en archivos ambientales.48 Las perturbaciones en el registro del plomo en los archivos naturales de las diversas regiones no son comparables en magnitud. Para el noroeste peninsular, desde los primeros estudios de metales contaminantes, como el plomo49 o el mercurio,50 se observa un máximo preindustrial entre los siglos I y II. Diversos trabajos posteriores en turberas han detectado un pico similar de contaminación en este periodo en 45 46 47 48 49 50 Murozumi et alii 1969; Hong et alii 1996. Martínez Cortizas et alii 2005. Bindler 2011. Renberg et alii 2001. Martínez Cortizas et alii 1997. Martínez Cortizas et alii 1999. López-Costas, O. Gerión, 39(2) 2021: 775-800 783 Galicia y Asturias.51 Un estudio de isótopos de plomo pudo determinar que dicha contaminación procedía de minas del norte, noroeste y suroeste de España, pero no de otras áreas de Europa como Reino Unido, Italia, Austria o Grecia.52 Este aumento en el plomo y mercurio muestra en ocasiones efectos sinérgicos con el contenido en arsénico, níquel o cadmio, lo cual es indicativo de que la contaminación es polimetálica y sugiere la explotación de una mayor diversidad de fuentes y procesos minero-metalúrgicos.53 Los archivos naturales de Galicia también coinciden en mostrar un moderado descenso en la contaminación a finales del siglo III inicios del siglo IV,54 mientras que en Asturias este descenso se retrasa hasta finales del siglo V, cuestión esta última que se ha vinculado a la explotación de minas de esta región.55 A partir del siglo V se observa una caída importante en la contaminación metálica en todo el noroeste peninsular,56 que se desacopla por primera vez de la deforestación.57 La explotación de los metales constituye una buena aproximación a la actividad humana en su vertiente más industrial, entendida esta por minería y metalurgia. Aunque la metalurgia puede ser desarrollada a pequeña escala para un uso familiar, lo cierto es que las grandes variaciones en su intensidad son probablemente una respuesta a la gestión socio-política de los recursos y no a decisiones individuales o al mero aumento de la población (ya que los metales no son recursos vitales). Fe de ello lo da la propiedad imperial de las minas en el mundo romano, que dependía directamente del emperador.58 Por ello, una caída de la actividad se podría relacionar con una menor estabilidad, lo que afectó a la explotación de estos lugares o la demanda de estos recursos. Es decir, que minería y metalurgia nutren principalmente a entidades supra-individuales, lo cual no deja de ser irónico cuando ahondamos en sus consecuencias. La contaminación metálica tiene consecuencias importantes para la salud, incluso a bajas dosis, debido a la intolerancia del cuerpo a metales tóxicos como el plomo, el mercurio o el arsénico, que acaban incorporándose y causando problemas orgánicos o sistémicos, así como acrecentando las posibilidades de que otras enfermedades resulten fatales. Quizás se nos venga a la cabeza la idea de mineros o personas directamente relacionadas con el metal y tremendamente contaminados. No obstante, la más importante, a mi modo de ver, es aquella contaminación latente que estaba siendo incorporada por la población en general incluso en áreas muy alejadas. Pensemos que si los metales llegaban a las turberas situadas en lo alto de las montañas, éstos estaban en el aire, la comida y bebida de las gentes. Por lo tanto, esta contaminación crónica de relativamente baja entidad es la más extensa. Recientemente he propuesto que los esqueletos (humanos) arqueológicos sean 51 52 53 54 55 56 57 58 Ej. Martínez Cortizas et alii 2002a; Martínez Cortizas et alii 2002b; Martínez Cortizas et alii 2013. Kylander et alii 2005. Pontevedra-Pombal et alii 2013; en este artículo se indica la contaminación polimetálica, lo cual interpreto como la explotación de varias fuentes o recursos. Kylander et alii 2005. Martínez Cortizas et alii 2013. Martínez Cortizas et alii 2002a; Martínez Cortizas et alii 2002b; Kylander et alii 2005; Martínez Cortizas et alii 2013; Pontevedra-Pombal et alii 2013. López-Merino et alii 2014, los autores lo sitúan en torno al 440-700 y relacionan el menor descenso en la deforestación con un cambio en el uso de la tierra y una mayor dependencia del pastoreo en esta zona de montaña, pero no de prácticas agrícolas ya que el polen de cereal está ausente. Zubiaurre Ibáñez 2017. López-Costas, O. Gerión, 39(2) 2021: 775-800 784 considerados archivos ambientales al mismo nivel que otros de los que he hablado anteriormente, ya que registran y almacenan variaciones de su entorno.59 Uno de los aspectos a los que más pueden aportar es el estudio de la contaminación metálica. El registro de los cambios temporales y la validez de los esqueletos como archivos de la contaminación se demuestra en el trabajo que venimos haciendo en el yacimiento de A Lanzada (Sanxenxo, Pontevedra). Se trata de una población rural alejada de los grandes centros de minería, donde se han podido identificar dos áreas de enterramiento, una que cubre desde el siglo I hasta mediados del IV y otra que va del siglo IV al VII. En estos trabajos hemos demostrado cómo la contaminación por plomo y mercurio, incorporada en los individuos durante su vida, tiene una intensidad y huella isotópica similar a la registrada en otros archivos ambientales, con valores característicos de contaminación metálica relativamente intensa en el primer periodo y más bajos en el segundo. Los resultados coinciden a la perfección con la señal obtenida en las turberas de las que hemos hablado anteriormente. Además, en el caso del mercurio, se detectó un patrón intra-esquelético que sugiere que dicho metal estaba almacenado en los llamados órganos diana, entre ellos el hígado. Las investigaciones que estamos haciendo en estos momentos se encaminan a identificar mejor este patrón, así como a ampliar el número de estudios para esta región del noroeste.60 Estos trabajos nos enseñan que la contaminación llegaba a todas las poblaciones, así como que la caída de la actividad minero-metalúrgica durante la Tardoantigüedad dio lugar, probablemente, a una mejora de la salud de las personas (al menos en este aspecto). 5. Una alimentación que se adapta La dieta en el noroeste de España parece presentar desde la Antigüedad unas características distintivas entre las que se incluye una dependencia del consumo de las denominadas plantas C4, que se remonta al Bronce Final, primeramente en forma de mijos y posteriormente de maíz.61 Los mijos son utilizados aquí directamente para el consumo humano. A su vez, y de acuerdo con los trabajos que hemos realizado, existe una marcada diferencia entre la alimentación de la costa y el interior, estando la primera caracterizada por un consumo extraordinariamente alto de productos marinos,62 mucho mayor que otras regiones europeas, costeras o no. Una de las dificultades de la aproximación bioarqueológica a la alimentación de este periodo es cómo conocer los cambios en la dieta con respecto a los anteriores y posteriores, desechando variaciones locales. Es decir, para los objetivos que nos proponemos no resulta especialmente ilustrativo acercarnos a la alimentación de una 59 60 61 62 Oliva et alii 2018; López-Costas et alii 2020. López-Costas et alii 2020, estudio de isótopos de plomo y concentración de plomo y mercurio; López-Costas et alii 2016, trabajo previo donde se diferencia la contaminación pre- y postmortem; Álvarez-Fernández et alii 2020, exploran la contaminación por mercurio en vida analizando diversos tipos de huesos. Esta metodología ha sido también aplicada a esqueletos de Suecia con idénticos buenos resultados. Para la discusión de la introducción de los mijos ver López-Costas et alii 2015; estos corresponden a panizo, Setaria italica (L.) P.Beauv., y, sobre todo, mijo común, Panicum miliaceum L. Para la introducción del maíz en la alimentación humana y animal ver López-Costas – Müldner 2019. López-Costas 2012; López-Costas – Müldner 2016; López-Costas – Müldner 2019, directamente en esqueletos humanos y huesos de animales, coincidentes para los recursos marinos con Grandal d’Anglade et alii 2015 y en el uso del mijo con Teira Brión 2019. López-Costas, O. Gerión, 39(2) 2021: 775-800 785 zona muy concreta, así como tampoco el reconstruirla para un momento específico sin llegar a conocer si se trata de una preferencia de ese periodo o una continuidad del anterior. En el caso de los estudios de restos botánicos o animales contrasta la relativa abundancia de yacimientos de periodo romano, mientras que el número de aquellos que abarcan más allá del siglo IV decrece drásticamente. Un ejemplo de ello lo aporta la revisión de los trabajos de carpología, que incluye veintiún yacimientos para el noroeste de la Península entre los periodos romano y medieval, de los cuales solo uno tiene muestras adscribibles a los siglos V-VII.63 Estos estudios de semillas y otros para Galicia64 revelan el consumo de varios cereales en el noroeste “romano”, tales como diversos trigos, avena, cebada, centeno y espelta. Esta última se mantiene en Galicia y en la montaña asturiana, pero desaparece de los yacimientos tardoantiguos y medievales de toda la Península. Se encuentran también sobradas evidencias de mijos, mijo común y panizo, importantes en la economía del noroeste donde constituyen el 30% de las semillas recuperadas (siendo de solo un 3% en otras áreas). Al menos para Asturias, parece que la espelta y otros tipos de escandas son predominantes en aquellas zonas que por su humedad y bajas temperaturas hacen difícil el cultivo de los mijos.65 La escasa variedad de legumbres, habas (en concreto Vicia faba L.), arvejas (Vicia sativa L.) y guisantes, contrasta con la del resto de la Península Ibérica. Otros alimentos vegetales encontrados en el noroeste incluyen castañas, nueces, avellanas, piñones, manzanas, cerezas, uvas, higos, aceitunas o nabos. En proporción, lo que más destaca son los cereales. No existen, por tanto, grandes diferencias que nos permitan hablar de un tipo de cultivos característicos de la Tardoantigüedad, aunque el consumo de cebada sí parece haber aumentado a partir del siglo IV. En relación con otras áreas peninsulares, en el noroeste destaca la importancia relativa que tuvieron los mijos, la cebada y la espelta, mientras que el centeno no despuntó hasta el periodo medieval.66 Semillas de ciruelas, melocotones, pepinos o melones fueron identificadas en un puerto costero de la actual ciudad de Vigo, por lo que pueden ser un reflejo del tráfico marítimo y no haber estado disponibles para toda la población.67 Incluso en las áreas habitacionales de nueva construcción dedicadas a explotaciones concretas, como la minería en la zona de Las Médulas (El Bierzo, León), se hallaron trigo, cebada, mijo y habas.68 Con respecto a la comida “ritualizada”, no tenemos evidencias de restos vegetales para el periodo estudiado, pero se han hallado guisantes, habas, lentejas y huesos de animales en piras funerarias del siglo I en el norte de Portugal, lo cual se relaciona con el ritual 63 64 65 66 67 68 Peña-Chocarro et alii 2019, analizan un total de 252 muestras (~1/5 del total para todos los periodos, todas las áreas), que son adscribibles cronológicamente a los siglos II y VII, la mayoría de contextos rurales. Existen, no obstante, 11 yacimientos que tienen alguna fase entre los siglos IV al VI. Condensados en la tesis Teira Brión 2019. Ver discusión sobre la convivencia entre mijos (verano) y escanda (invierno) y las grandes áreas dedicadas al cultivo de esta última en la montaña asturiana (Vigaña) en Fernández Mier 1999, 268. Ver también Fernández Mier 2013. Peña-Chocarro et alii 2019. La abundancia de restos carpológicos de frutas exóticas en el depósito anegado de las salinas de Vigo –Teira Brión 2013– contrasta por ejemplo con la escasez de restos carpológicos de importancia económica de otros registros más alejados de zonas de tráfico marítimo como el pozo-depósito Tardoantiguo (siglos VI-VIII) de la fábrica de tabacos de Gijón, ver Carrión Marco et alii 2015. López-Merino et alii 2010, referido a los trabajos realizados en el yacimiento romano de Castro de Orellán, figs. 6 y 11. El trigo se refiere aquí a trigo farro (Triticum dicoccum L.). 786 López-Costas, O. Gerión, 39(2) 2021: 775-800 funerario.69 A mi conocimiento, no existen hallazgos similares o huesos de animales atribuibles a este uso en ninguna de las inhumaciones de Galicia, aunque sí se han encontrado ajuares cerámicos.70 Con respecto a la cabaña ganadera, el periodo romano no parece suponer en esta región un cambio drástico en las especies, dominando el ganado caprino (ovejas y cabras) mientras que el vacuno parece ser más frecuente en la zona cantábrica, siendo los cerdos menos frecuentes. Esta continuidad también se observa en el suroeste peninsular, aunque la caza del ciervo parece ser una actividad más importante aquí.71 En las zonas urbanas o aquellas más romanizadas de nueva planta, el ganado porcino adquiere una mayor importancia, llamando la atención los huesos de cochinillos, de aves (gallinas y gansos) y de caza.72 Destacan también hallazgos puntuales de camélidos (dromedarios) en estratos tardíos de Astorga.73 La cabaña ganadera del noroeste sufre también transformaciones en cuanto a los morfotipos, que se vuelven ahora más grandes y con mayor aprovechamiento de carne, aunque se mantienen los animales de menor tamaño típicos de la Edad de Hierro. También aparecen nuevos tipos de caballo y perros pequeños asociados al ámbito doméstico de las clases pudientes. Teniendo en cuenta el volumen, el ganado vacuno parece ser el que proporcionaba mayor cantidad de carne a la dieta, aunque el caprino posiblemente aumentó a partir del siglo IV.74 Sin embargo, la escasez de yacimientos pertenecientes a la Tardoantigüedad no nos permite conocer si hubo modificaciones severas en el tipo de cabañas ganaderas ni en su tamaño corporal, tal y como se constató en otras áreas del norte peninsular.75 En el mundo romano, con respecto a la Edad de Hierro, se observan ligeros cambios en la pesca, ya que las artes introducidas facultan la captura de especies de fondo y bancos de jureles o sardinas en mar abierto. Aumentaron y se diversificaron las fábricas de salazón, ya presentes en el periodo anterior, aunque ahora se procesan especies como la sardina, posibilitando la llegada de productos marinos elaborados hasta las zonas interiores.76 Los peces más representados son la sardina, la dorada, la maragota y el jurel, aumenta la presencia de abadejo y merluza, y decrecen en importancia la faneca y el besugo.77 Se han encontrado restos de erizos y ostras en localidades del interior, como Astorga. Existió una distribución de los moluscos en 69 70 71 72 73 74 75 76 77 Vaz et alii 2020; las legumbres tienen marcas compatibles con haber sido cocidas, sin embargo, otros cereales están ausentes aquí. Los huesos de animales no pudieron ser identificados salvo un fragmento de radio de cerdo joven (< 4 años). Se han encontrado ajuares cerámicos en tumbas claramente adscribibles al siglo IV en Asturias (Requejo Pagés 2015) y posiblemente adscribibles a la primera mitad de este siglo en A Lanzada (López-Costas 2015; LópezCostas et alii 2020). Valenzuela-Lamas – Detry 2017; estas autoras encuentran también un paulatino aumento en el tamaño de los cerdos y una gran diversidad de prácticas ganaderas, aunque ha de tenerse en cuenta la gran amplitud de la región estudiada en este trabajo. Rodríguez 2000; Fernández Rodríguez 2003; Fernández Rodríguez – Fuertes Prieto 2007; se pueden encontrar buenas revisiones en estos trabajos para el Noroeste de España. Fernández Rodríguez – Fuertes Prieto 2007. Fernández Rodríguez – Fuertes Prieto 2007. Grau-Sologestoa 2015, quien ha estudiado la biometría de cabras-ovejas, vacas y cerdos, encontrando una reducción del tamaño de cerdos y cabras-ovejas en la Tardoantigüedad, sin que se vea afectado el tamaño de las vacas hasta el siglo VIII. Rodríguez Martínez et alii 2011, documentan una de estas explotaciones prerromanas en A Lanzada (Sanxenxo, Pontevedra). Para periodo romano destaca el yacimiento de Adro Vello (O Grove, Pontevedra) donde se encontraron seis piletas de salazón con vértebras y escamas de sardina y jureles (Ferré Álvarez 2003). González Gómez de Agüero 2013, 355-357, para la ictiofauna. López-Costas, O. Gerión, 39(2) 2021: 775-800 787 los periodos Alto y Bajoimperial (hasta el siglo V), los de roca en los asentamientos costeros y los bivalvos de arena en el interior, incluyendo grandes ciudades como Lugo o Astorga. En estas aparecen también abundantes restos de ostras y vieiras, fiel retrato de un comercio de recursos marinos destinado muy posiblemente a las élites.78 Un elemento necesario para la realización de salsas y salazones es la sal, que adquiere importancia industrial en el sur de la actual Galicia a partir del siglo I. Estudios realizados en salinas de la zona de Vigo, área intensamente explotada desde el cambio de era, indican el desuso de la mismas a partir del siglo V y hasta el VII,79 relacionado con modificaciones en las corrientes y dinámica de la ría. Los estudios bioarqueológicos no permiten ir más allá de este ejemplo concreto. Lo mismo sucede con otros productos, los diversos autores asumen un cambio, si no en las especies, sí en la intensidad de su explotación, relacionado con la pérdida de estructura comercial que parece que sigue al siglo IV; sin embargo, todos resaltan la falta de yacimientos. Hemos de indicar aquí que los estudios arqueobotánicos y zooarqueológicos son importantísimos para conocer la dieta a nivel poblacional, aunque no es posible conocerla para una persona o grupo concreto. Además, en estos casos tenemos que ser conscientes de que las condiciones de conservación pueden generar sesgos interpretativos. Es decir, el hecho de que la mayoría de los materiales botánicos estudiados estén conservados por carbonización, y muchos menos por saturación de agua, podría causar la sobrerrepresentación de aquellas semillas o maderas con más posibilidades de tener contacto con el fuego y no ser calcinadas, sino carbonizadas. En el caso de los restos animales, y al igual que los humanos, normalmente llegan a nosotros solo los esqueletos (incluyendo conchas). Un modo directo de reconstruir la dieta en el pasado lo proporciona el análisis de los restos óseos humanos empleando metodologías como la patología oral o los análisis de isótopos estables en hueso y diente. Galicia destaca por la abundancia de estudios de isótopos estables, los cuales nos permiten aproximarnos a la alimentación de hombres, mujeres y niños por separado, o enlazar esta información con otra para asumir el estrato social. Los estudios de isótopos estables para la reconstrucción de la dieta han revelado un alto consumo de mijos tanto en el interior (yacimiento de Ouvigo, Os Blancos, Ourense) como en la costa (yacimientos de A Lanzada, O Grove, Pontevedra; y Calle Real, A Coruña).80 Estos mijos no eran usados como forraje del ganado, como sucede en otras áreas peninsulares, sino consumidos directamente por las personas. Los yacimientos costeros muestran un característico patrón de moderado-alto consumo de recursos marinos. A partir del siglo IV el consumo de recursos marinos y mijos aumenta considerablemente, constituyendo probablemente la base de la dieta. Este cambio de patrón ha sido observado en la necrópolis de A Lanzada, cuyas características únicas de contar con dos áreas funerarias me permiten profundizar sobre cómo cambió la alimentación en una misma área. Para Galicia, hemos observado también un aumento similar o incluso mayor del consumo de recursos marinos y mijo (o incluso maíz) durante periodos posteriores de deterioro 78 79 80 Bejea García 2015, 570-577, para la malacofauna mencionada. Tallón-Armada et alii 2018. López-Costas 2012; Silva-Sánchez et alii 2013, datos de los esqueletos romanos de Ouvigo (siglos II-IV). López-Costas – Müldner 2016, para la reconstrucción de la dieta de A Lanzada. Grandal d’Anglade et alii 2015 y Kaal et alii 2016 para la Calle Real de A Coruña. 788 López-Costas, O. Gerión, 39(2) 2021: 775-800 climático como la Pequeña Edad del Hielo.81 Una mayor dependencia de los mijos en la alimentación humana y animal se detectó en otros yacimientos europeos, y en todos los casos lo hemos asociado a un empeoramiento de las condiciones climáticas a partir del siglo IV, que podrían haber dificultado el cultivo de otros cereales, así como todas las características de cambios ambientales y sociales, y los movimientos de población.82 Las modificaciones en la alimentación de A Lanzada no coinciden con un tipo de tumba o ajuar concreto de los enterramientos, y tampoco ha podido identificarse ningún elemento que ayude a la adscripción religiosa de ningún individuo. Hemos propuesto, incluso, que dichos cambios pudieron estar promovidos de alguna manera por la llegada de los grupos germánicos, ya que los mijos eran consumidos en la zona actualmente ocupada por Alemania. Sin embargo, los estudios isotópicos y carpológicos nos hablan de que dichos cultivos eran de sobra conocidos y empleados en el noroeste peninsular desde al menos el Bronce Final. Es, por tanto, difícil conocer si el cambio de alimentación hacia una mayor dependencia del mar y de las plantas C4 se debe a un condicionamiento externo ambiental, principalmente social o a una combinación de ambos. Con seguridad, las futuras investigaciones sobre otras necrópolis de esta área, así como sobre la alimentación infantil, nos ayudarán a ahondar en estos interrogantes. 6. Breves apuntes sobre la morfología, salud, enfermedad y costumbres funerarias El examen de otros aspectos de la vida y muerte de los individuos, así como si estas cambiaron a partir del siglo IV, depende del análisis directo de los restos esqueléticos humanos, no especialmente abundantes en esta región.83 Sin embargo, estos estudios son dependientes de un buen tamaño muestral ya que se requiere de una mayor interpretación a nivel poblacional.84 Esto no quiere decir que en los casos anteriores de reconstrucción de la dieta y de la contaminación la perspectiva poblacional no sea necesaria, ni que en estos últimos el estudio de individuos o grupos pequeños no tenga importancia. Cuando se trabaja con la llamada geoquímica del esqueleto (ej. dieta por isótopos estables) es posible obtener un dato de cada uno de los esqueletos casi independientemente de cómo de completo esté el mismo. En cambio, en otros trabajos, como la antropometría, la paleopatología o la tafonomía, se hace necesario que una determinada pieza esté completa, si no todo el esqueleto, reduciendo la cohorte de individuos que se pueden estudiar. Por ello, la fortaleza de las conclusiones que se extraen muchas veces es limitada. Nos vamos de nuevo al yacimiento de A Lanzada, donde no se han observado grandes diferencias en el morfotipo y antropometría entre los esqueletos recuperados en las áreas funerarias romana (siglos I-IV) y tardoantigua (siglos IV-VII). Las 81 82 83 84 Oliva et alii 2018; López-Costas – Müldner 2019; definimos la Pequeña Edad de Hielo como el periodo del 1300 al 1850. López-Costas – Müldner 2016; discutimos estos aspectos, por ejemplo para Alemania Hakenbeck et alii 2010. Posteriormente Jordana et alii 2019 retoman esta cuestión observando un ligero aumento para Cataluña. Debate sobre el uso de mijos en la Península Ibérica en López-Costas – Alexander 2019. Solo siete yacimientos con esqueletos estudiados se pueden adscribir a los siglos estudiados; para un resumen de estudios de osteoarqueología en época medieval para Galicia consultar López-Costas – Sánchez Pardo 2016, en concreto la figura 2.1 y la tabla 2.1. Para caracterizar una población (estadística), entendida esta aquí como una cohorte de individuos que comparten características, ha de analizarse un número de muestras representativo de la misma. López-Costas, O. Gerión, 39(2) 2021: 775-800 789 estaturas, tanto de la serie femenina como de la masculina, son similares, dando en su conjunto una media de 157±5 cm y 165±5 cm, respectivamente.85 Ha de tenerse en cuenta que si quisiésemos comparar estos individuos con otras poblaciones deberíamos emplear los valores estimados de estatura, evitando contrastar las medidas concretas de un hueso.86 Las estaturas estimadas en A Lanzada son mediaaltas para los hombres y altas en las mujeres cuando se comparan con las de otras poblaciones de la Península. Debido a la elevada estatura media de estas últimas, el dimorfismo sexual no es muy acusado. Un bajo dimorfismo sexual se relaciona con malas condiciones de vida, ya que ambos sexos estarían sometidos a un fuerte estrés en la infancia, lo cual no les permitiría alcanzar la estatura potencial máxima. En un análisis de diversas poblaciones peninsulares se observa un aumento en la robustez y la estatura media en el periodo romano y tardoantiguo, hecho también observado en Galicia.87 Sin embargo, el hecho de que ambas series de A Lanzada tengan valores supra-medios con respecto a otras series españolas, hace que no nos podamos decantar por esta hipótesis basada en el dimorfismo sexual.88 En base al estudio paleopatológico de los esqueletos arqueológicos datados entre los siglos IV y VII sabemos que esta población estaba en contacto con enfermedades infecciosas bacterianas, como la tuberculosis y la brucelosis. He hallado marcas compatibles con dos posibles casos crónicos de tuberculosis y otros dos de brucelosis en la segunda fase de A Lanzada,89 pero ninguna en la primera ya que en su excavación no se recogieron los huesos cortos y las marcas patognomónicas de ambas patologías se encuentran en la columna vertebral.90 Estas patologías dejan marca en el esqueleto solo en los casos crónicos, por lo que no es de extrañar que los diagnosticados conviviesen con bastantes más casos agudos, hecho que sugiere una extensión importante de ambas enfermedades en la población. Estas enfermedades estaban también presentes en otras poblaciones del periodo de la Península y no parece existir un aumento en la Tardoantigüedad.91 Aunque algunos autores han sugerido la existencia de una “involución” en las prácticas médicas de este periodo con respecto a la época clásica, no se han encontrado evidencias claras para Galicia.92 Finalmente, muchos de estos cambios se reflejan tanto en la vida como en la muerte. En un trabajo sobre la arqueología funeraria de las dos fases de A Lanzada se determinó que la fase romana se caracteriza por una orientación de las tumbas Norte-Sur y Sur-Norte, ajuar y enterramientos con ropa. Se incluyen también en esta fase cremaciones (excavadas, pero no recogidas) y enterramientos en decubito prono con el cuerpo depositado con poco cuidado y, en un caso, posiblemente 85 86 87 88 89 90 91 92 López-Costas 2012, 197. Las series están compuestas por 10 muestras en cada caso. Estimación realizada en base a Pearson 1899. En población viva se observa que personas con la misma estatura tienen medidas óseas diferentes (por ejemplo, de la longitud máxima del fémur) dependiendo de aspectos como el ángulo de inserción en la cadera, las cuales se pueden estimar de manera bastante aceptable si empleamos fórmulas que atiendan a estas variaciones a nivel poblacional. Un ejemplo de lo que creo que es un mal uso metodológico, donde se han comparado medidas concretas en vez de la estimación de la estatura empleando las fórmulas específicas de cada población, se puede observar en Harper 2017, 75-78. Estudio biométrico realizado por Jordana – Malgosa 2004; ver también discusión en Menéndez Bueyes 2013, 124, basado sobre todo en la tesis de Varela López 1974-75. Para Galicia véase López-Costas et alii 2007. López-Costas 2012, 201-203, comparación con 22 poblaciones de entre los siglos I al XVIII. López-Costas 2012. Aufderheide – Rodríguez-Martín 1998. Menéndez Bueyes 2013, 91-96; Menéndez Bueyes 2016. Menéndez Bueyes 2013, 119. 790 López-Costas, O. Gerión, 39(2) 2021: 775-800 maniatado. Los enterramientos presentan piezas constructivas de barro cocido como tégulas o ímbrices, o se hicieron directamente sobre el terreno. En la segunda área, la tardoantigua, el ajuar desaparece por completo, así como los restos arqueológicos que sugieran piezas de vestir o calzado. La orientación cambia a un estricto Oeste-Este y se ha anotado la presencia de conchas cerca de los cuerpos, aunque nuestras últimas intervenciones en el yacimiento nos hacen pensar que pudiesen ser depósitos casuales ya que la necrópolis está en una duna.93 La posición de los cuerpos es en decubito supino y aparecen enterramientos múltiples no consecutivos en tumbas de lajas de piedra, lo cual indica la reutilización de las mismas. Una reutilización similar la hemos observado en una reciente excavación en la calle Oliva de Vigo, en una de las pocas tumbas adscribibles estratigráficamente a la Antigüedad Tardía.94 En el caso de los individuos colocados boca abajo del periodo imperial, la falta de ajuar y el hecho de que se depositaran en un área funeraria al lado de otros que muestran una posición no desviada no nos permite determinar si existió una actitud punitiva o de rechazo; las características antropológicas tampoco proporcionan información de si se trataba de personas libres o esclavas. En la segunda fase encontramos un esqueleto cuyos huesos del pie muestran marcas de mordiscos de un cánido en hueso fresco, este tratamiento descuidado podría asociarse a un mismo fenómeno de enterramientos depositados en áreas anómalas a partir del siglo V en Cataluña,95 o ser un hallazgo puntual. Hasta donde conozco no hay casos similares descritos, lo cual también puede deberse a falta de estudios detallados sobre marcas tafonómicas. Finalmente, he discutido la posible adscripción de estas prácticas funerarias a un cambio de creencia, como la llegada del cristianismo,96 por ejemplo, sin haber encontrado evidencias claras, por lo que dejaré esta discusión a otros que puedan llevarla más allá de la evidencia material a la que me acojo. Evidentemente, no es posible determinar la adscripción religiosa de las personas inhumadas en este periodo, ni el registro bioaqueológico nos permite conocer sus vínculos, puramente hipotéticos, con el priscilianismo. 7. La vida en tiempos de Prisciliano y de los priscilianistas, una reflexión Los priscilianistas y sus coetáneos vivieron en una Galicia surcada por profundas modificaciones. Reducirlas aquí a un único factor, el cambio religioso, sería una 93 94 95 96 En las primeras excavaciones (Blanco Freijeiro et alii 1967) se indica la presencia de conchas, de berberecho, almeja y en un caso de vieira, en las tumbas 16, 18, 19, 22, 23 y 30, estas son identificadas como parte del ajuar. Aparecen cerca de diversas zonas del esqueleto (entre las piernas, al lado de un brazo, de una pierna, cerca de la cabeza) y algunas son ejemplares no completos. Una consulta de las fotografías de la intervención (es difícil adscribirlas a una tumba concreta; véase López-Costas 2015) no revela una intencionalidad ni protocolización en la colocación de dichas conchas, y estas aparecen en las tumbas excavadas en la duna, no en las realizadas en otros suelos. En la intervención de 2016-2017 observamos la presencia de conchas de especies similares en la arena de la duna, fuera y dentro de los enterramientos, por lo que parece poco probable que se trate de un ajuar, sino más bien de depósitos no intencionales relacionados con el material de partida donde se realizó el enterramiento. Habría que revisar otros yacimientos costeros para poder ahondar en las sobreinterpretaciones de este tipo de hallazgos. López-Costas 2015, para la arqueología funeraria en contexto con el estudio de los restos humanos. El estudio de la calle Oliva está en curso. López-Costas 2015, para la descripción de los enterramientos fuera de norma en A Lanzada. Olesti Vila 2019, expone un interesante resumen de los esqueletos hallados en áreas no funerarias o en posiciones anómalas en el noreste peninsular. López-Costas – Müldner 2016. López-Costas, O. Gerión, 39(2) 2021: 775-800 791 gran simplificación, tal y como he querido mostrar en las líneas anteriores. Es muy posible que las gentes se vieran condicionadas por la inestabilidad derivada de la imposición de una nueva élite que parece haber convivido de manera agitada con la vieja, pero que también pacta con ella tal y como demuestra la paz firmada en el año 433 entre los suevos y los Gallaeci.97 Sin embargo, ¿cómo podemos ignorar la realidad que impuso el clima? Sin caer en un determinismo ambiental simplista, se debe admitir que el empeoramiento de las condiciones de humedad y temperatura sin duda condicionó la vida de las gentes del campo, así como la selección de los cultivos más apropiados, de entre los que se conocían en el mundo romano. En este caso no intento imponer una visión de crisis total ni negar la opulencia de los grandes terratenientes del siglo IV.98 Mi intención es dejar patente que las condiciones de vida se pudieron ver alteradas por los cambios ambientales, especialmente las de las clases más humildes, sin caer en un catastrofismo ambiental. Dicho esto, los tiempos manejados por los archivos ambientales no se ajustan a la vida de una generación de personas, por lo que quizás las modificaciones climáticas que parecen cruciales a escala de centurias no tuviesen un impacto definitivo en las vidas individuales (aunque sí no ignorable). Un aspecto sobre el que me gustaría reflexionar aquí, pues ha sido uno de los principales focos de mi investigación en los últimos años, es el del papel jugado por los mijos. Este tipo de cultivos, conocidos como cereales de primavera, se puede plantar a finales de esta estación o en verano, recogiendo el grano en un par de meses. Resultan una opción ideal si se quiere dedicar los terrenos a pasto el resto del año, o como contrapunto al más delicado trigo o a otros cereales de invierno como la cebada. La resistente espelta también puede ser una buena opción ante el empeoramiento climático, tal y como constatan los estudios realizados en la montaña asturiana. Son, por lo tanto, los mijos y la espelta, combinados con la ganadería en el interior y los recursos marinos en la costa, buenas opciones para momentos de inestabilidad/cambio social y ambiental. Su aumento entre los siglos IV y VI está, creo yo, vinculado a esos cambios. Si esta modificación en las preferencias se relaciona con las nuevas creencias es difícil de decir; quizás el hecho de que ambos sucedieran a la vez proporcionó un contexto doctrinal a la nueva realidad que cada vez se alejaba más del utópico mundo romano, como tanto nos recuerda Hidacio. Las fuentes documentales y las bioarqueológicas difieren en que las primeras se centran en asuntos puntuales de marcado impacto social, mientras que las otras nos hablan de aquellos aspectos que formaban parte del día a día. En nuestros estudios no hemos hallado evidencias ni de violencia interpersonal ni de pestes. El saqueo y las masacres sucedidas en ciudades como Asturica Augusta o Bracara Augusta99 no tienen un reflejo en los restos óseos; si bien es verdad que estos eventos que crean gran alarma social (como el asesinato de un obispo en su iglesia)100 no tuvieron por qué extenderse a toda la población ni mucho menos llegar a zonas rurales como las 97 98 99 100 Hyd. Chron. 100; López Barja de Quiroga 1999. Brown 2012. Hyd. Chron. 183 (Asturica Augusta), 167 y 173 (Bracara Augusta); Burgess 1989, 150: hambre y episodios de canibalismo en las ciudades saqueadas. Candelas Colodrón 2004, 245 y 247, para el saqueo de Braga y para la destrucción de Astorga y de sus campos de cultivo, respectivamente. Hyd. Chron. 141, con Burgess 1989, 219: Leo obispo de Tyriasso en 449. 792 López-Costas, O. Gerión, 39(2) 2021: 775-800 estudiadas. Las pestes mencionadas por Hidacio,101 las cuales son golpes previos a la gran peste de Justiniano (541-549), no tienen un reflejo en el registro óseo. Tampoco lo tienen otros eventos como los terremotos de los años 451 y 454.102 No es esperable que lo tuviesen ya que, de causar muertes, la rapidez de ambos fenómenos impide la formación de marcas en los esqueletos, restringiéndose las evidencias a la presencia de fosas comunes, las cuales, de existir, no han sido aún descubiertas. Podrían ser evidencias sutiles el descuido en el cuidado del cuerpo de los difuntos observado ocasionalmente en A Lanzada y otras áreas de la Península, el cual sí se puede relacionar con la inmediatez, así como con grupos en movimiento o muertes que sucedieron de manera imprevista. Se ha debatido sobre el catastrofismo en la obra de Hidacio asumiendo que adquiere un grado de irrealidad importante. Según algunos autores a partir de un momento en que la Chronica versa sobre “la destrucción total del Reino Suevo de la Gallaecia”103 este discurso se vuelve tan alejado de la realidad que “podemos eliminar la reconstrucción histórica que se refiere a la destrucción de las ciudades”. Dicho supuesto se basa principalmente en la mención a la presencia de camellos durante la destrucción de Bracara como parte del ejército de los godos.104 En concreto, Javier Arce indica que, en su opinión, “estos camellos nunca existieron”, pues lo ve como algo imposible y una exageración del cronista, y en ello basa su argumentación de catastrofismo irreal. No obstante, los estudios bioarqueológicos han revelado la existencia de huesos de camélidos (dromedarios) en Astorga coincidentes con el periodo narrado.105 Carlos Fernández Rodríguez y Natividad Fuertes Prieto hacen una excelente descripción de dichos hallazgos. Dicho esto, no cabe duda de que Javier Arce tiene razón cuando señala que el siglo V fue “un siglo en transición de unas estructuras políticas, administrativas, económicas, una concepción de la vida de la ciudad y de creencias religiosas, pero marcadas en muchos aspectos por la continuidad”.106 En este sentido apunta también la información bioarqueológica. Pero, a su vez, sí creo que hubo momentos puntuales de caos y destrucción, los cuales se vieron magnificados en la vida de la gente que los sufrió y dejaron un impacto en una Chronica que quizás no es tan irreal como pensamos, al menos en lo que se refiere a la presencia de camélidos en la Gallaecia. La posible inestabilidad puntual concuerda con el uso de las cuevas como zona habitacional provisional y una vuelta a la ocupación de áreas elevadas que proporcionan buen refugio. Es posible, entonces, que el descenso de la cobertura arbórea después de la ligera recuperación del siglo IV se relacione con esta nueva manera de vivir en respuesta a la inseguridad puntual de los grandes 101 102 103 104 105 106 Hyd. Chron. 126; Burgess 1989, tabla 4. Burgess 1989, tabla 4. Cita literal de Arce 1995, 219, quien expresa su incredulidad sobre la crónica de Hidacio en base a la presencia de camellos en la Gallaecia. Esta cita ha sido usada por autores como Díaz (2000) o Candelas Colodrón (2004), entre otros muchos, para justificar la visión irreal y catastrófica dada en la Chronica. Literalmente: iumentorum, pecorum, camellorumque horrore locus sacer impletus (Hyd. Chron. 174); Burgess 1989. Esto es interpretado por Arce 1995, como que estos animales forman parte del ejército del rey godo Teodorico, sin embargo lo que se indica en la crónica es que estos animales entraron en el lugar sagrado (locus sacer impletus) sin especificar si pertenecían o no a dicho ejército. Estos hallazgos se explican en Fernández Rodríguez – Fuertes Prieto 2007, señalando que se trata de un periodo tardío (con respecto al periodo romano) y que posiblemente se relacionen con movimientos de tropas, quizás desde el Norte de África o empleados para el comercio y carga de bienes. Reflexiones incluidas en Arce 2013, 281-284. López-Costas, O. Gerión, 39(2) 2021: 775-800 793 núcleos de población. Las gentes trasladan la agricultura, pero sobre todo la ganadería, a estas zonas para subsistir, mientras que la minero-metalurgia ya no es una actividad esencial del día a día. Hechos que tendrían como consecuencia el observado desacoplamiento de la contaminación metálica y la deforestación. Los robledales probablemente tuvieron que acoger a las gentes que se trasladan de los núcleos más poblados, movimiento quizás alentado por las hambrunas de las que habla Hidacio, como la que siguió a la entrada en Hispania de los suevos, vándalos y alanos.107 No podría asegurar si estos movimientos responden a un aumento de la fe eremítica, pero de ser así constituirían seguramente una buena manera de “ganarse el pan” en este periodo. A la vez, estos modos de vida podrían ser el germen de los cenobios, que tan intensamente popularizaría San Fructuoso de Braga en la Gallaecia. La información bioarqueológica indica un cierto aumento de la presión antrópica sobre el medio a partir del siglo V, después de la caída del siglo IV. Si esto constituye un aumento de la actividad o un crecimiento económico a nivel de toda Galicia es un interesante debate al cual aún no podemos aportar suficientes datos. Además, ¿cómo podríamos explicar la escasez de yacimientos de esta cronología que los autores que he citado indican?108 Por otro lado, quizás estos hechos estén relacionados con un aumento de la producción autosuficiente y la economía autárquica que se vuelve más familiar asociada al retraimiento económico y a una nueva manera de producir,109 que, creo yo, pudiesen resultar más adecuadas a las nuevas condiciones ambientales. Lo que sí parece claro es que el tipo de actividades cambió siendo la mayor explotación del bosque y superficie de prados aspectos clave. Aparece una nueva manera de vivir que pudiera estar mejor adaptada a un peor clima y al aumento de la erosión intensificada por un desbroce incontrolado, de ahí los sistemas de terrazas. No se puede interpretar esto directamente como un aumento demográfico, ya que el impacto antrópico sobre el medio no depende del número de habitantes sino de las actividades que éstos desarrollan. Quizás esta nueva realidad se adapte mejor a la existencia de unas nuevas élites locales que ya no dependían de Roma, pero sus características son difíciles de abordar desde este tipo de estudios. La bioarqueología nos va a hablar más del conjunto de la población, y muy rara vez de las personas que ostentaban el poder. Finalmente, los priscilianistas de esta región, así como sus congéneres, posiblemente se enfrentaron a una modificación de las condiciones socioambientales que pudieron causar, hasta cierto punto, una ruptura en la resiliencia de su entorno y formas de vida. Su legado se puede ver no solo en las costumbres funerarias, sino también en los complejos sistemas de terrazas o socalcos, y en los brezales, todos ellos tan característicos del paisaje gallego actual. Animo aquí a que otros investigadores empleen esta información para abordar aspectos diferentes de la historia social y económica, y juntos podamos añadir nuevas perspectivas al conocimiento que tenemos de estos siglos. 107 108 109 Hyd. Chron. 48; Burgess 1989, 91, se refiere al hambre y enfermedad del ganado, quizás pudiésemos relacionarlo con la brucelosis, enfermedad eminentemente del ganado que afecta también a los humanos. Pudiendo tratarse también de malas asignaciones cronológicas. Sobre la reflexión del cambio en la economía y la productividad Arce 2013, 275-279. Brown 2012, 139-162 incide también en la importancia de las élites locales cada vez más alejadas de Roma. Nótese que en estos trabajos el uso de la información bioarqueológica es prácticamente nulo, por ejemplo buscando referencias concretas al tipo de ganado consumido en la Chronica de Hidacio antes que en los estudios zooarqueológicos Arce 2013, 276. 794 López-Costas, O. Gerión, 39(2) 2021: 775-800 8. 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https://openalex.org/W2995015739	https://hal.archives-ouvertes.fr/hal-03091586/document	English	null	Quantitative and qualitative analysis of asynchronous neural activity	bioRxiv (Cold Spring Harbor Laboratory)	2,019	cc-by	12,971	Quantitative and qualitative analysis of asynchronous neural activity Ekkehard Ullner, Antonio Politi, Alessandro Torcini ehard Ullner, Antonio Politi, Alessandro Torcini To cite this version: Ekkehard Ullner, Antonio Politi, Alessandro Torcini. Quantitative and qualitative analysis of asyn- chronous neural activity. Physical Review Research, 2020, 2 (2), 10.1103/PhysRevResearch.2.023103. hal-03091586 I. INTRODUCTION even for relatively small network connectivities [5,6]; (ii) the non-Poissonian nature of the spiking activity; (iii) large amplitude of the single spikes and the consequential possible failure of a perturbative, linear approach; and (iv) the presence of non-negligible ﬁnite-time correlations. The characterization of the spiking activity of neuronal networks is a long-standing problem even with reference to the asynchronous regime: simple from a dynamical point of view, but extremely relevant for understanding cortex dynam- ics [1,2]. A moment’s reﬂection indeed suggests that this is not a trivial task whenever the self-generated input current is not constant: To what extent can the deterministic ﬂuctuations be treated as a stochastic process? Several alternative approaches have indeed been proposed. For instance, an exact treatment of shot noise for spike ampli- tudes not vanishingly small, which leads to a mixed Fokker- Planck/master-equation formalism (see Refs. [7,8]). Unfortu- nately, we are not aware of any way to make the approach self-consistent, by inferring the input properties on the basis of the observed output. Anyhow, since this approach assumes a Poissonian distribution of the interspike intervals (ISIs)—a property largely unsatisﬁed for strong synaptic coupling—one should look for different approximation schemes anyway. Generally, the problem is tackled by assimilating the input current to a δ-correlated Gaussian process [3] and thereby deriving a self-consistent Fokker-Planck equation. Under the additional simplifying assumption of a piecewise linear dynamics [as in leaky integrate-and-ﬁre (LIF) neurons], a solution can be analytically determined and its stability assessed [4]. A different strategy was proposed by Dummer et al. [9], based on the self-consistent derivation of the power spectrum of the spiking activity. The advantage of this method is that no assumption is made on the spectral shape of the synaptic current. While the original implementation already proved unstable for relatively small coupling strengths, the variant recently proposed in Ref. [10] leads to a seemingly accurate reproduction of the network dynamics. We shall treat it as a reference for some of our considerations. Although this approach turns out to be relatively accurate for small coupling strengths, the same is no longer true for stronger coupling (see Ref. [5]), when large deviations from the theoretical predictions are observed. Ekkehard Ullner and Antonio Politi tute for Pure and Applied Mathematics and Department of Physics (SUPA), Old Aberdeen, Aberdeen AB24 3UE, United Kingdom Ekkehard Ullner and Antonio Politi Institute for Pure and Applied Mathematics and Department of Physics (SUPA), Old Aberdeen, Aberdeen AB24 Alessandro Torcini Laboratoire de Physique Théorique et Modélisation, Université de Cergy-Pontoise, CNRS, UMR 8089, 95302 Cergy-Pontoise cedex, France and Istituto dei Sistemi Complessi, CNR—Consiglio Nazionale delle Ricerche, via Madonna del Piano 10, I-50019 Sesto Fiorentino, Italy (Received 17 December 2019; accepted 25 March 2020; published 29 April 2020) (Received 17 December 2019; accepted 25 March 2020; published 29 April 2020) The activity of a sparse network of leaky integrate-and-ﬁre neurons is carefully revisited with reference to a regime of bona ﬁde asynchronous dynamics. The study is preceded by a ﬁnite-size scaling analysis, carried out to identify a setup where collective synchronization is negligible. The comparison between quenched and annealed networks reveals the emergence of substantial differences when the coupling strength is increased, via a scenario somehow reminiscent of a phase transition. For sufﬁciently strong synaptic coupling, quenched networks exhibit a highly bursting neural activity, well reproduced by a self-consistent approach, based on the assumption that the input synaptic current is the superposition of independent renewal processes. The distribution of interspike intervals turns out to be relatively long-tailed; a crucial feature required for the self-sustainment of the bursting activity in a regime where neurons operate, on average, (much) below threshold. A semiquantitative analogy with Ornstein-Uhlenbeck processes helps validate this interpretation. Finally, an alternative explanation in terms of Poisson processes is offered under the additional assumption of mutual correlations among excitatory and inhibitory spikes. DOI: 10.1103/PhysRevResearch.2.023103 HAL Id: hal-03091586 https://hal.science/hal-03091586v1 Submitted on 31 Dec 2020 Submitted on 31 Dec 2020 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. PHYSICAL REVIEW RESEARCH 2, 023103 (2020) Published by the American Physical Society DOI: 10.1103/PhysRevResearch.2.023103 A. Network model where τ = 20 ms is the membrane time constant, RI0 = 24 mV is an external DC supra-threshold “current,” while RIi is the synaptic current arising from the recurrent coupling RIi(t) = τJ n Gi j(n)δ t −t ( j) n −τd , (2) (2) where J is the coupling strength and the sum runs over all the spikes emitted at time t ( j) n < t from the presynaptic neurons j(n) connected to neuron i. Gi j is the adjacency matrix and its elements assume the following values: Gi j = 1 (−g), if the presynaptic neuron j is excitatory (inhibitory), otherwise Gi j = 0. If Vj reaches the threshold Vth = 20 mV at time t ( j) n , two events are triggered: (i) the membrane potential Vj is reset to Vr = 10 mV and held ﬁxed for a refractory period τr = 0.5 ms; (ii) a spike is emitted and received τd = 0.55 ms later by the postsynaptic cells connected to neuron j according to Gi j. Except for the system size N, all parameters are set as in Ref. [11]: b = 0.8, K = 1000, and g = 5, so each neuron receives input from bK [(1 −b)K] excitatory (inhibitory) presynaptic neurons. Besides this quenched setup, we have considered annealed networks, where the postsynaptic neigh- bors are randomly chosen at each spike emission. As a matter of fact, in the former (latter) case, the in degree (out degree) is equal to K for each neuron, while the out degree (in degree) is binomially distributed with average K and standard deviation √ K, in the thermodynamic limit. Our choice was dictated by efﬁciency of the numerical simulations; however, we have veriﬁed that no substantial changes are observed if, instead of ﬁxing the number of links equal to K, the probability of each link is set equal to c = K/N, as in truly Erdös-Renyi networks. One of the main results of this paper is that the synap- tic current can be accurately represented as the superposi- tion of independent identical renewal processes (RPs), each characterized by a suitable ISI distribution. We also show that the correlations due to the long-tailed ISI distribution can be equivalently represented as long-term memory in the symbolic representation of inhibitory versus excitatory spikes. p More precisely, in Sec. A. Network model The ﬁrst goal is to determine the minimal network size such that the numerical results can be considered as truly asymptotic. This is achieved by ﬁrst checking the accuracy of numerical simulations and then estimating ﬁnite-size cor- rections. In Refs. [5,6], it was indeed shown that the ﬁring activity of a network of 10 000 LIF neurons with an in-degree K = 1000 is strongly affected by the presence of collective irregular dynamics. Due to its relevance in the context of asynchronous dynam- ics in balanced networks [4,11,18,19], we consider the follow- ing sparse spiking network of LIF neurons. The network is composed of bN excitatory and (1 −b)N inhibitory cells; the membrane potential Vi of the ith neuron evolves according to the equation τ ˙Vi = R(I0 + Ii) −Vi , (1) (1) g y To clarify the role played by the different “ingredients,” we introduce a second setup: annealed networks, where post- synaptic neighbors are randomly selected whenever a new spike is emitted. From a biological point of view, this setup can be justiﬁed by recalling that synaptic transmission in the cortex is highly unrealiable: On average, at room temperature, most synapses respond to only ≃13% of the the presynaptic spikes [13]. Unreliable synapses are usually analyzed at either the single-unit level [14], or of neural populations [15] to understand how this unreliability actually affects information transmission. Moreover, from a statistical-mechanics point of view, this choice will help in better understanding the dynamics of the system, since it is known that quenched and annealed systems may exhibit different properties from one another [16]. Interestingly, this happens in the present model above a “critical” synaptic coupling strength (namely, J ≈ 0.25). The existence of two seemingly different phases was already claimed by Ostojic [11], but challenged in Ref. [17]. Here, however, we do not investigate the behavior in the vicinity of the hypothetical phase transition, but rather focus on the large-coupling regime, since it is more important to ﬁrst characterize the different phases which may emerge upon varying the coupling strength. I. INTRODUCTION These deviations may in principle originate from various sources: (i) the sponta- neous onset of irregular collective dynamics, which emerges In this paper, we revisit the problem of characterizing the asynchronous regime in the context of two coupled popula- tions of excitatory and inhibitory LIF neurons accompanied by refractoriness and delay [4,11,12]. In spite of the restriction to a speciﬁc type of neurons (motivated by the opportunity to compare with many numerical and analytical studies), the Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. 023103-1 023103-1 2643-1564/2020/2(2)/023103(12) PHYSICAL REVIEW RESEARCH 2, 023103 (2020) ULLNER, POLITI, AND TORCINI B. Methods A detailed description of network dynamics requires look- ing both at the microscopic and the macroscopic level. A. Network model II, we introduce the model and deﬁne the indicators later used to characterize and discuss the various dynamical properties. In the following Sec. III, we illustrate the ﬁring activity of the quenched network, computing several indicators for different coupling strengths. A relatively quick discussion is also devoted to the annealed set-up to show the differences with respect to the quenched case. In Sec. IV, we ﬁrst introduce the two self-consistent approaches herein implemented to characterize the neural activity. The former one, based on the distribution of ISIs, provides a rather accurate description. The latter, already proposed in Refs. [9,10], reveals an unexpectedly stable ﬁxed point, which, however, is further away from the results of accurate simulations. In Sec. V, we turn our attention to the bursting activity observed for large coupling in an attempt to explain how neurons operating, on average, below threshold are able to exhibit a strong ﬁring activity. Finally, in Sec. VI we summarize the main results and focus on the still open problems. II. MODEL AND METHODS approach we propose is general and can be implemented regardless of the single-neuron evolution equation. 1. Microscopic indicators The dynamics of a spiking neuron is usually characterized in terms of the probability distribution function (PDF) Q(T ) of the ISIs T and of the associated moments: namely, the mean ISI T and the standard deviation σT of T . Usually, the regularity/irregularity of the dynamics is quantiﬁed by the so-called coefﬁcient of variation, Cv = σT T , equal to zero for a periodic dynamics and to 1 for Poissonian spike trains. It should be noted that Cv can be larger than 1 for the so-called bursting dynamics, when the neuron alternates 023103-2 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) QUANTITATIVE AND QUALITATIVE ANALYSIS OF 0 20 40 60 80 integration time [s] 30 40 50 60 ν [Hz] _ FIG. 1. Running average of the ﬁring rate in a network of N = 105 neurons with a coupling strength J = 0.8 and overall connectivity K = 103. The black solid line refers to event-driven simulations; the dashed lines correspond to Euler integration scheme with different time steps (δt = 0.1 ms, 0.04 ms, 0.01 ms, 0.001 ms, from bottom to top). 0 20 40 60 80 integration time [s] 30 40 50 60 ν [Hz] _ periods of silence and high activity. The ﬁring rate of a neuron is simply given by ν = 1/T . To characterize the network activity we estimate the mean coefﬁcient of variation ⟨Cv⟩ and the mean ﬁring rate ⟨ν⟩, where ⟨·⟩represents an ensemble average over all neurons. An important observable is the power spectrum S( f ) = ⟨\|˜u( f )\|2⟩, where ˜u( f ) with f = m/(Mδt) is the Fourier trans- form of the neural activity u(t), determined by computing the number of spikes emitted in M consecutive time intervals of duration δt. Further observables we focused on are the phase correlations among different frequencies f and h, which can be quantiﬁed by the normalized indicator, D( f ) = h̸= f ⟨\|˜u(h) ˜u∗( f )\|2⟩ W S( f ) , (3) (3) where W = ⟨ h \|˜u(h)\|2⟩is the total power of the spectrum. One can check that 0 ⩽D( f ) ⩽1, the lower (upper) bound corresponding to uncorrelated (perfectly correlated) channels. The typical values we have used in our simulations are δt = 0.11 ms and M = 215 (M = 212) for the power spectrum (phase correlations) estimation. FIG. 1. 2. Macroscopic indicators We then proceed to analyzing the dependence of the av- erage ﬁring rate ⟨ν⟩on the coupling strength J. The black, solid curve in Fig. 2(a) has been obtained for N = 104 and exactly the same parameter values as in Ref. [11]. As reported therein, ⟨ν⟩, after an initial drop, increases with the coupling strength J. One of the goals of this paper is to understand the origin of this growth in a network where inhibition is expected to prevail over excitation. A theoretical estimate νT of the average ﬁring rate in the asynchronous regime of a sparse network can be derived from the stationary solution of a self-consistent Fokker-Planck equation, under the assumption of an uncorrelated Poissonian activity of the neurons [3,4]. However, this prediction, based on the diffusion approxima- tion [3] and reported as a dotted green curve in Fig. 2(a), is able to reproduce only the initial part of the curve ⟨ν(J)⟩, while it fails to describe the growth observable for larger coupling. Furthermore, in Refs. [5,6] it was found that the corresponding dynamical phase is far from asynchronous; this is testiﬁed by the behavior of ρ(J), reported in Fig. 2(b), where we can see that the order parameter ρ can be as large as 0.5. Considering that the theoretical prediction has been derived under the assumption of a strictly asynchronous regime (i.e., ρ = 0), it is therefore crucial to separate out the effects of the collective dynamics. Deviations from stationarity reveal the presence of a col- lective dynamics. To measure the level of coherence in the network dynamics, a commonly used order parameter is [21] ρ2 ≡⟨v⟩2 −⟨v⟩ 2 ⟨v2 −v2⟩ , (4) (4) where the overbar denotes a time average. In practice, ρ is the rescaled amplitude of the standard deviation of the average ⟨v⟩. When all neurons behave in exactly the same way (perfect synchronization), the numerator and the denominator are equal to one another and ρ = 1. If instead they are independent as in an asynchronous regime, ρ ≈1/ √ N due to the central limit theorem. 1. Microscopic indicators Running average of the ﬁring rate in a network of N = 105 neurons with a coupling strength J = 0.8 and overall connectivity K = 103. The black solid line refers to event-driven simulations; the dashed lines correspond to Euler integration scheme with different time steps (δt = 0.1 ms, 0.04 ms, 0.01 ms, 0.001 ms, from bottom to top). 2. Macroscopic indicators event driven scheme. This is indeed the value employed in our simulations performed with Euler’s scheme. Notice that for δt = 0.1 ms, a value often chosen in the literature, the ﬁring rate is substantially underestimated (by approximately 24%). We then proceed to analyzing the dependence of the av- erage ﬁring rate ⟨ν⟩on the coupling strength J. The black, solid curve in Fig. 2(a) has been obtained for N = 104 and exactly the same parameter values as in Ref. [11]. As reported therein, ⟨ν⟩, after an initial drop, increases with the coupling strength J. One of the goals of this paper is to understand the origin of this growth in a network where inhibition is expected to prevail over excitation. A theoretical estimate νT of the average ﬁring rate in the asynchronous regime of a sparse network can be derived from the stationary solution of a self-consistent Fokker-Planck equation, under the assumption of an uncorrelated Poissonian activity of the neurons [3,4]. However, this prediction, based on the diffusion approxima- tion [3] and reported as a dotted green curve in Fig. 2(a), is able to reproduce only the initial part of the curve ⟨ν(J)⟩, while it fails to describe the growth observable for larger coupling. Furthermore, in Refs. [5,6] it was found that the corresponding dynamical phase is far from asynchronous; this is testiﬁed by the behavior of ρ(J), reported in Fig. 2(b), where we can see that the order parameter ρ can be as large as 0.5. Considering that the theoretical prediction has been derived under the assumption of a strictly asynchronous regime (i.e., ρ = 0), it is therefore crucial to separate out the effects of the collective dynamics. At the mean-ﬁeld level, the network evolution is captured by the instantaneous PDF P(v,t) of the membrane potentials of the neurons. In the limit case of an inﬁnitely large in- degree, the perfectly asynchronous regime is characterized by a constant ﬁring rate ⟨ν⟩[20]. This implies that the ﬂux of neurons along the v axis is independent of both time and potential value, i.e., the corresponding PDF P0(v) should be stationary. event driven scheme. This is indeed the value employed in our simulations performed with Euler’s scheme. Notice that for δt = 0.1 ms, a value often chosen in the literature, the ﬁring rate is substantially underestimated (by approximately 24%). A. The quenched network 0 50 100 200 250 T [ms] 10 -6 10 -5 10 -4 10 -3 10 -2 Q(T) (a) 200 400 800 1000 T [ms] 10 -5 10 -4 10 -3 10 -2 10 -1 Q(T) 10 0 10 1 10 3 T [ms] 10 -4 10 -2 Q(T) (b) 0 0.2 0.4 0.8 J 0 10 20 30 40 50 <ν> [Hz] 0 0.2 0.4 0.8 J 0 0.2 0.4 ρ 0 0.2 0.4 0.8 J 0 1 2 3 4 <Cv> 0.001 0.01 0.1 c 0.1 1 ρ (b) (d) (c) (a) FIG. 2. Collective properties of the network dynamics versus the coupling strength J: (a) average ﬁring rate ⟨ν⟩, (b) the coherence order parameter ρ, (c) the mean coefﬁcient of variation ⟨Cv⟩. Solid lines and symbols refer to quenched networks (namely, black lines correspond to N = 104, blue ones to N = 105, and (red) crosses to N = 8 × 105) (blue) dashed lines correspond to simulations of the annealed network performed for N = 105 neurons. The (green) dotted line in panel (a) corresponds to the theoretical prediction νT derived in Refs. [3,4]. Finally, panel (d) displays the coherence order parameter ρ versus the connectivity c = K/N, reporting only results for the quenched network for different synaptic coupling: namely, J = 0.1, J = 0.5, and J = 0.8 (solid lines from the bottom to the top). The dotted line refers to the theoretical scaling law √c. 0 0.2 0.4 0.8 J 0 0.2 0.4 ρ 0.001 0.01 0.1 c 0.1 1 ρ (b) (d) 0 50 100 200 250 T [ms] 10 -6 10 -5 10 -4 10 -3 10 -2 Q(T) (a) (b) 0 0.2 0.4 0.8 J 0 1 2 3 4 <Cv> (c) 10 -3 10 -2 10 -1 Q(T) 10 0 10 1 10 3 T [ms] 10 -4 10 -2 Q(T) (b) FIG. 2. Collective properties of the network dynamics versus the FIG. 2. Collective properties of the network dynamics versus the coupling strength J: (a) average ﬁring rate ⟨ν⟩, (b) the coherence order parameter ρ, (c) the mean coefﬁcient of variation ⟨Cv⟩. A. The quenched network Solid lines and symbols refer to quenched networks (namely, black lines correspond to N = 104, blue ones to N = 105, and (red) crosses to N = 8 × 105) (blue) dashed lines correspond to simulations of the annealed network performed for N = 105 neurons. The (green) dotted line in panel (a) corresponds to the theoretical prediction νT derived in Refs. [3,4]. Finally, panel (d) displays the coherence order parameter ρ versus the connectivity c = K/N, reporting only results for the quenched network for different synaptic coupling: namely, J = 0.1, J = 0.5, and J = 0.8 (solid lines from the bottom to the top). The dotted line refers to the theoretical scaling law √c. 500 1000 1500 T [ms] 10 -6 10 -5 10 -4 10 -3 10 -2 10 -1 Q(T) 1 10 1000 T [ms] 10 -6 10 -4 10 -2 Q(T) (c) collective effects increase as ρ ≈√c, consistent with theo- retical expectations [4]. Going back to Fig. 2(a), we see that upon further increasing N above 105, the ﬁring rate for a given coupling strength J saturates to a ﬁnite value. Altogether, we can safely conclude that the increase of ν for J ≳0.3 is a genuine property of bona ﬁde asynchronous activity and should be explained as such. Our simulations suggest that the system size N = 105 is large enough to ensure nearly asymptotic results and small enough to allow for affordable simulation times. From now on, all simulations will refer to this network size, unless stated otherwise. The main questions we want to address are understanding (i) the features of such a high ﬁring-rate regime and (ii) why it deviates so strongly from the diffusion approx- imation [3,4], even for a not-too-large synaptic coupling J in a setup where correlations among the different neurons are practically absent. FIG. 3. The PDF Q(T ) of the ISIs T generated by the full quenched network (black) and as obtained after the ﬁrst step of the renewal process recursive procedure (red curve). Panels (a)–(c) refer to J = 0.1, J = 0.5, and J = 0.8, respectively. The insets contain the same information in log-log scales to emphasize the initial quasi- power-law decay. The panel (c) inset shows additionally in green the output of a single neuron subject to a symbol-correlated Poisson process (see Sec. V C). that a theoretical approach, such as that in Refs. [3,4], based on the assumption that ⟨Cv⟩= 1, is not accurate. A. The quenched network Our simulations have mostly been performed by imple- menting an exact event-driven scheme [22,23]; see Ref. [6] for a description of the details. Since, however, some simu- lations required implementing a clock-driven Euler scheme and since this latter approach is often used in the literature, we have ﬁrst compared the two algorithms for a network of 105 neurons with a coupling strength J = 0.8. From the results reported in Fig. 1, we see that the time step δt used in the implementation of Euler’s algorithm should not be larger than 10−3 ms to get results essentially in agreement with the This can be done by increasing the network size while leaving the in-degree ﬁxed (namely, K = 1000). Quite sur- prisingly, the ﬁring rate obtained for N = 105 [see the up- per blue solid curve in Fig. 2(a)] displays an even more pronounced growth than for N = 104, in spite of a weaker synchronization, as shown in Fig. 2(b). The analysis re- ported in Fig. 2(d), where ρ vs c = K/N is reported for three different synaptic coupling values, shows that the 023103-3 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) ULLNER, POLITI, AND TORCINI PHYSICAL REVIEW RESEARCH 2, 023103 (2020) 0 50 100 200 250 T [ms] 10 -6 10 -5 10 -4 10 -3 10 -2 Q(T) (a) 200 400 800 1000 T [ms] 10 -5 10 -4 10 -3 10 -2 10 -1 Q(T) 10 0 10 1 10 3 T [ms] 10 -4 10 -2 Q(T) (b) 500 1000 1500 T [ms] 10 -6 10 -5 10 -4 10 -3 10 -2 10 -1 Q(T) 1 10 1000 T [ms] 10 -6 10 -4 10 -2 Q(T) (c) FIG. 3. The PDF Q(T ) of the ISIs T generated by the full quenched network (black) and as obtained after the ﬁrst step of the renewal process recursive procedure (red curve). Panels (a)–(c) refer to J = 0.1, J = 0.5, and J = 0.8, respectively. The insets contain the same information in log-log scales to emphasize the initial quasi- power-law decay. The panel (c) inset shows additionally in green the output of a single neuron subject to a symbol-correlated Poisson process (see Sec. V C). that a theoretical approach, such as that in Refs. [3,4], based on the assumption that ⟨Cv⟩= 1, is not accurate. Cv gives only rough information about the distribution of the ISIs. It is worth turning our attention to the full shape of the ISI distribution Q(T ). In Fig. 3, we plot Q(T ) for J = 0.1, J = 0.5, and J = 0.8; in all cases, we see that for large enough A. The quenched network Before proceeding along these lines, it is useful to pro- vide a more detailed description of the network activity. In Fig. 2(c), the mean coefﬁcient of variation ⟨Cv⟩is plotted versus J for different networks sizes. There, we see that Cv steadily increases with J and converges to some asymptotic value upon decreasing c. In practice, the neural activity can never be treated as a Poissonian process, as requested by the diffusion approximation employed in Ref. [4]; it is either more regular (for small coupling) or substantially more intermittent (as for J > 0.4). Therefore, it should not come as a surprise that a theoretical approach, such as that in Refs. [3,4], based on the assumption that ⟨Cv⟩= 1, is not accurate. Cv gives only rough information about the distribution of the ISIs. It is worth turning our attention to the full shape of the ISI distribution Q(T ). In Fig. 3, we plot Q(T ) for J = 0.1, J = 0.5, and J = 0.8; in all cases, we see that for large enough that a theoretical approach, such as that in Refs. [3,4], based on the assumption that ⟨Cv⟩= 1, is not accurate. Cv gives only rough information about the distribution of the ISIs. It is worth turning our attention to the full shape of the ISI distribution Q(T ). In Fig. 3, we plot Q(T ) for J = 0.1, J = 0.5, and J = 0.8; in all cases, we see that for large enough 023103-4 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) QUANTITATIVE AND QUALITATIVE ANALYSIS OF 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (a) 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (b) 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (c) FIG. 4. Power spectra S( f ) of the neural activity. Black curves refer to the full quenched network for N = 105; red curves refer to the ﬁrst step of the renewal process approximation. Panels (a)– (c) refer to J = 0.1, 0.5, and 0.8, respectively. A. The quenched network 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (a) 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (b) ISIs, the PDF is characterized by an exponential tail as for a Poissonian process. However, for small ISIs, the PDF is substantially different. For weak coupling, very small ISIs are strongly inhibited [24]: This is an obvious consequence of the nearly constant input current (mean driven). For stronger couplings, the PDF exhibits a quasi-power-law decay which extends up to 10–20 ms [see the insets of Figs. 3(b) and 3(c)]. These features are consistent with the characterization of the ISI distributions reported in Ref. [25] for spiking neurons driven by ﬂuctuating inputs. In particular, the PDF shown in Fig. 3(a) is expected to emerge when the average effective input current (including the contribution of the synaptic cou- pling) lies between the threshold and the reset value, which is indeed the case. Furthermore, we have computed the power spectrum S( f ) of single spike sequences. For weak coupling (J = 0.1), a small peak is visible at f = 17 Hz in Fig. 4(a) [26] it is reminiscent of the periodic activity of the uncoupled neurons. At higher frequencies, the spectrum is practically ﬂat, i.e., it is approximately white. Upon increasing the coupling, the spectrum starts exhibiting a low-frequency peak, suggesting the presence of “long”-time correlations. This feature will be further discussed in Sec. V with reference to the emergence of a bursting activity. For J = 0.8, subsidiary peaks related to the delay emerge for f = 1818 Hz and its multiples. The delay is always present but for unexplained reasons pops up only for large coupling when the white-noise background is even larger. 10 0 10 1 10 2 10 3 f [Hz] 10 1 10 2 10 3 S(f) [Hz] (c) B. The annealed network So far, we focused on the dynamical properties of a net- work characterized by a quenched distribution of synaptic connections. However, all theoretical approaches developed to characterize the ﬁring activity do not take into account the actual, invariant structure of the connections. Even more, theoretical approaches do not include delay at all. Therefore, it is natural to ask to what extent the quenched nature of the network may be considered responsible for the observed asynchronous activity. This question can be addressed by considering an annealed network, where the “neighbors” of each given neuron are randomly assigned each time a spike is emitted. More precisely, we proceed as follows: Whenever a neuron ﬁres, we still assume that the quality of the spike (excitatory versus inhibitory) is determined by the neuron itself, but we randomly choose K receiving neurons regardless of their quality. Moreover, we exclude self-connections, i.e., the sender must differ from the receiver. Finally, we keep all parameters as in the quenched network. This guarantees that, on average, each neuron receives bK excitatory inputs and (1 −b)K inhibitory ones. Annealed neural networks are usually employed to mimic the effect of unreliable synapses present in the cortex [15,27]. 10 3 FIG. 4. Power spectra S( f ) of the neural activity. Black curves refer to the full quenched network for N = 105; red curves refer to the ﬁrst step of the renewal process approximation. Panels (a)– (c) refer to J = 0.1, 0.5, and 0.8, respectively. assumes an annealed connectivity. An additional justiﬁcation for this agreement is the Cv values reported in Fig. 2(d) (see the dashed curve) which are much smaller than in the quenched case and closer to 1, the value corresponding to a Poisson process. The numerical results for the average ﬁring rate are re- ported in Fig. 2(a). There we observe a good agreement with the behavior of the quenched network for J ≲0.3, while increasing deviations emerge for larger coupling strengths. Interestingly, the behavior of the annealed setup is very close to the theoretical prediction νT [4]. This is not entirely surpris- ing since, as anticipated, the theoretical approach implicitly A. Renewal process In practice, we have implemented the following recursive procedure: given the ISI distribution Qk(T ) determined in the kth step, we have generated the synaptic current RI of a generic neuron [in the (k + 1)st step] by superposing K independent RPs all built according to the same distribution Qk(T ) (under the constraint that bK spikes are excitatory and the remaining ones inhibitory). Upon afterward integrating the single-neuron equation, we have generated the ﬁring activity induced by the current RI, thereby determining the (k + 1)st distribution Qk+1(T ). eventually drives it away along the unstable manifold. If, in analogy to what was done in Ref. [10] for the Gaussian approximation, we include memory effects by building the new ISI PDF as the average of the last two distributions, the saddle is stabilized: The resulting ﬁxed point is represented in Fig. 5 as Pr. The nonperfect correspondence between Pq and Pr may have a double rationale: the RP assumption is not exact; the network size used to determine Pq is not large enough. We ﬁrst focused on J = 0.8, since the theoretical pre- diction νT [3] is signiﬁcantly inaccurate for this coupling strength. The initial condition Q0(T ) has been selected as the distribution generated by a quenched network of N = 105 neurons with an in-degree K = 1000. The corresponding ﬁring rate is ⟨ν⟩= 50.4 Hz and its coefﬁcient of variation is ⟨Cv⟩= 3.97 [29]. This pair of values is represented by the point Pq in Fig. 5: It corresponds to the projection of the asynchronous state of the quenched network in this two- dimensional space. The iterates of the recursive procedure have been projected on the same plane; they are so close to each other to be hardly discernible in the main panel (see the enlarged plot presented in the upper inset of Fig. 5 for a clearer representation). The closeness among consecutive iterates is conﬁrmed by the shape of the ISI distribution: In Fig. 3(c), we see that Q1(T ) is practically indistinguishable from Q0(T ). Altogether, these observations strongly hint at the existence of a ﬁxed point of the RP recursive procedure in the vicinity of Pq. IV. SELF-CONSISTENCY In the previous section, we have seen that quenched and annealed networks behave in a substantially different way, when the coupling strength is larger than J = 0.25. To what 023103-5 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) ULLNER, POLITI, AND TORCINI extent is this difference the signature of the crucial role played by a ﬁxed structure of the synaptic connections? 0 10 20 30 50 60 70 ν [Hz] 0 1 2 3 4 5 6 Cv 48 49 50 ν [Hz] 3.85 3.90 3.95 Cv 70 72 74 ν [Hz] 4.85 4.9 Cv Pa Pq Pq Pr Ps Pr Ps FIG. 5. Results of the recursive procedure based on the renewal process and the Gaussian approximation. Pq and Pa identify the dy- namics of quenched and annealed networks, respectively. Red circles (green diamonds) refer to the iterates of the RP iterative procedure while starting from Pa (Pq); green diamonds are better visible in the enlargement reported in the upper inset, where Pr denotes the ﬁxed point of the renewal process approximation. Blue squares refer to the iterative process based on the Gaussian approximation: Pq is the initial condition, while Ps denotes the ﬁxed point of this approach (see the lower inset for an enlargement of the later stages). All data have been obtained for J = 0.8). 0 10 20 30 50 60 70 ν [Hz] 0 1 2 3 4 5 6 Cv 48 49 50 ν [Hz] 3.85 3.90 3.95 Cv 70 72 74 ν [Hz] 4.85 4.9 Cv Pa Pq Pq Pr Ps Pr Ps In this section, we address this issue by implementing a self-consistent approach, where the input current is assumed to be the superposition of independent signals, each sharing the same “statistical” properties of the single-neuron activity. Two different approximation schemes are hereby discussed: (i) the hypothesis of a perfect RP and (ii) mutually uncorre- lated frequency channels (also termed Gaussian approxima- tion). Below we show that the former one provides a more accurate representation of the neural activity. A. Renewal process A RP is fully characterized by the ISI probability distribu- tion Q(T ). Assuming Q(T ) is known, a typical spike sequence can be readily generated by randomly drawing a series of T values according to this distribution. At variance with Ref. [9], where the authors suggested the idea of approximating the synaptic current with a RP, here we limit ourselves to assume that the single-neuron output activity is a RP, but we do not extend the assumption to the input, which is treated as the superposition of K independent RPs. This is an important difference since, as already remarked in Ref. [28], the super- position of RPs is not renewal itself unless the single processes are purely Poissonian (this is not our case). So, at variance with Ref. [9], we relax the condition of a strictly renewal input process and replace this ansatz with the more general hypothesis of a superposition of independent RPs. FIG. 5. Results of the recursive procedure based on the renewal FIG. 5. Results of the recursive procedure based on the renewal process and the Gaussian approximation. Pq and Pa identify the dy- namics of quenched and annealed networks, respectively. Red circles (green diamonds) refer to the iterates of the RP iterative procedure while starting from Pa (Pq); green diamonds are better visible in the enlargement reported in the upper inset, where Pr denotes the ﬁxed point of the renewal process approximation. Blue squares refer to the iterative process based on the Gaussian approximation: Pq is the initial condition, while Ps denotes the ﬁxed point of this approach (see the lower inset for an enlargement of the later stages). All data have been obtained for J = 0.8). FIG. 5. Results of the recursive procedure based on the renewal process and the Gaussian approximation. Pq and Pa identify the dy- namics of quenched and annealed networks, respectively. Red circles (green diamonds) refer to the iterates of the RP iterative procedure while starting from Pa (Pq); green diamonds are better visible in the enlargement reported in the upper inset, where Pr denotes the ﬁxed point of the renewal process approximation. Blue squares refer to the iterative process based on the Gaussian approximation: Pq is the initial condition, while Ps denotes the ﬁxed point of this approach (see the lower inset for an enlargement of the later stages). All data have been obtained for J = 0.8). A. Renewal process The black curve corresponds to the spectrum of the activity of a single LIF neuron driven within the RP approach (it is basically indistinguishable from the true quenched network activity). The red curve is obtained by randomizing the phases of the input signal. Data refers to J = 0.8. 1 10 100 1000 f [Hz] 0 0.01 0.02 D FIG. 6. Degree of phase correlations D( f ) of the Fourier trans- form of the spiking activity for J = 0.8. The lowermost blue line cor- responds to the RP; the uppermost black solid line corresponds to the activity of the quenched network. Finally, the red line corresponds to the annealed network. 10 0 10 1 10 2 10 3 f [Hz] 10 2 10 3 S(f) [Hz] FIG. 7. Power spectra for the network activity. The black curve corresponds to the spectrum of the activity of a single LIF neuron driven within the RP approach (it is basically indistinguishable from the true quenched network activity). The red curve is obtained by randomizing the phases of the input signal. Data refers to J = 0.8. 1 10 100 1000 f [Hz] 0 0.01 0.02 D FIG 6 Degree of phase correlations D( f ) of the Fourier trans 10 0 10 1 10 2 10 3 f [Hz] 10 2 10 3 S(f) [Hz] FIG. 7. Power spectra for the network activity. The black curve corresponds to the spectrum of the activity of a single LIF neuron driven within the RP approach (it is basically indistinguishable from the true quenched network activity). The red curve is obtained by randomizing the phases of the input signal. Data refers to J = 0.8. FIG. 6. Degree of phase correlations D( f ) of the Fourier trans- form of the spiking activity for J = 0.8. The lowermost blue line cor- responds to the RP; the uppermost black solid line corresponds to the activity of the quenched network. Finally, the red line corresponds to the annealed network. comparably small level of correlations is also found in the annealed network (see the almost ﬂat red line). a ﬁxed point of the RP iterative procedure. The reason is that while the temporal correlations exhibited by the single-neuron activity (encoded in the bursting activity) are preserved by the RP approximation, they are lost in the annealed setup because of the random reshufﬂing of the synaptic connections. A. Renewal process Further iterates start separating from each other, suggesting that the ﬁxed point is a saddle, which initially attracts the trajectory along the stable manifold and To test the quality of the RP approximation, we have studied the correlations of the sequence Tn of consecutive ISIs by estimating the so-called serial correlation coefﬁcient [30]: C(m) = ⟨Tn+mTn⟩−⟨Tn⟩2 T 2 n −⟨Tn⟩2 . (5) (5) In a strictly RP, C(m) = 0 for m ⩾1. Tests made on the neurons of a quenched network for N = 105 show that C(1) is at most of order 10−3, suggesting that the neural activity is well approximated by an RP. On the other hand, since the order parameter ρ is still relatively large for N = 105 (ρ = 0.17), ﬁnite-size effects are probably the predominant source of differences between Pq and Pr. In a strictly RP, C(m) = 0 for m ⩾1. Tests made on the neurons of a quenched network for N = 105 show that C(1) is at most of order 10−3, suggesting that the neural activity is well approximated by an RP. On the other hand, since the order parameter ρ is still relatively large for N = 105 (ρ = 0.17), ﬁnite-size effects are probably the predominant source of differences between Pq and Pr. What if the same recursive procedure is applied, starting from the dynamical regime exhibited by the annealed network (see point Pa in Fig. 5). Forward iterates rapidly move away from Pa and approach Pr (see the full circles in Fig. 5). The increasing amplitude of the “transversal” ﬂuctuations conﬁrm that Pr is a saddle point. Furthermore, the recursive procedure shows that Pa—a ﬁxed point of the annealed process—is not 023103-6 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) QUANTITATIVE AND QUALITATIVE ANALYSIS OF … 1 10 100 1000 f [Hz] 0 0.01 0.02 D FIG. 6. Degree of phase correlations D( f ) of the Fourier trans- form of the spiking activity for J = 0.8. The lowermost blue line cor- responds to the RP; the uppermost black solid line corresponds to the activity of the quenched network. Finally, the red line corresponds to the annealed network. 10 0 10 1 10 2 10 3 f [Hz] 10 2 10 3 S(f) [Hz] FIG. 7. Power spectra for the network activity. A. Renewal process The separation between Pa and Pr implicitly suggests the important role played by the bursting activity as will be conﬁrmed in the following section. As a second test of the relevance of phase correlations, we have investigated the consequence of phase randomization within the RP procedure. More precisely, given the synaptic current u(t) and its Fourier transform ˜u( f ), we have generated a new transform ˜uN( f ) = \|˜u( f )\|eiφ( f ) by randomly assigning the phase φ( f ) to the frequency f . A new signal uN(t) is then obtained by back transforming ˜uN( f ). The resulting spectrum of the ﬁring activity of a neuron subject to the current uN(t) is presented in Fig. 7 (see the red curve). The difference with the original spectrum (see the lower black curve) is not entirely negligible: It is around 20% in the low frequency region. Finally, we have also implemented the RP approach for smaller J values, always ﬁnding evidence of a weakly unsta- ble ﬁxed point (actually, the degree of instability decreases upon decreasing J). The resulting message is that the stable asynchronous dynamics exhibited by the quenched network is well reproduced by an unstable ﬁxed point of a recursive transformation based on the RP approximation. Finally, we have implemented the recursive procedure proposed in Ref. [9], here brieﬂy recalled. Let S(o) k ( f ) denote the power spectrum of the single-neuron spiking activity at the kth recursive step. Also, let S(i) k+1( f ) denote the power spectrum of the synaptic current in the (k + 1)st recursive step. In the asynchronous regime, the synaptic current is the superposition of K independent signals [bK excitatory and (1 −b)K inhibitory], all characterized by the same spectrum S(o) k ( f ). Taking into account the amplitude of the single spikes, we have that S(i) k+1 = [J2K(b + (1 −b)g)]S(o) k ( f ) [32]. The deﬁnition of the procedure is completed by adding the “rule” to generate S(o) k+1, given S(i) k+1( f ). This is done by feeding a single neuron with a phase-randomized spectrum (see the paragraph above). The self-consistent solution is ﬁnally iden- tiﬁed by the condition S(o) k+1 = S(o) k . V. BURSTING ACTIVITY In the same ﬁgure, we also report the outcome of annealed- network dynamics (see the crosses): for small R⟨I⟩, i.e., for small coupling we see an almost perfect coincidence. On the other hand, by further decreasing the internal current (i.e., upon increasing the coupling strength), the ﬁring rate does not increase in the annealed network, conﬁrming the qualitatively different behavior exhibited by the two setups. The annealed network operates above threshold. In the previous sections, we have seen that for strong coupling the neural activity is characterized by a large Cv > 1, a typical indication of bursting activity. This is indeed quali- tatively conﬁrmed in Fig. 8, where a raster plot is reported of the activity of a few neurons: Irregular bursts of spikes intermingled by long period of silence are clearly visible. Some neurons seem to be completely silent but this is just the consequence of the ﬁnite length of the time window (all neurons ﬁre with the same rate). In this section, we discuss in more quantitative terms this form of asynchronous dynamics, starting from the basic question of how it is possible for it to be self-sustained. In the previous sections, we have seen that for strong coupling the neural activity is characterized by a large Cv > 1, a typical indication of bursting activity. This is indeed quali- tatively conﬁrmed in Fig. 8, where a raster plot is reported of the activity of a few neurons: Irregular bursts of spikes intermingled by long period of silence are clearly visible. Some neurons seem to be completely silent but this is just the consequence of the ﬁnite length of the time window (all neurons ﬁre with the same rate). In this section, we discuss in more quantitative terms this form of asynchronous dynamics, starting from the basic question of how it is possible for it to be self-sustained. B. Power spectrum The vertical dashed line separates the region where neurons operate above (right) from below (left) threshold. The green crosses report the outcome of the annealed network. The two crosses deviating from the solid line belong to J = 0.5 and 0.8 in the annealed setup. -15 -10 -5 0 R<I> [mV] 0 20 40 60 80 <ν> [Hz] J=0.8 J=0.5 J=0.25 FIG. 9. Firing rate ⟨ν⟩versus the synaptic “current” R⟨I⟩for different coupling strengths for the quenched network (solid line and red circles). Zero coupling corresponds to the rightmost point, where the synaptic current obviously vanishes. The vertical dashed line separates the region where neurons operate above (right) from below (left) threshold. The green crosses report the outcome of the annealed network. The two crosses deviating from the solid line belong to J = 0.5 and 0.8 in the annealed setup. 0 50 100 150 200 t [ms] 0 20 40 60 80 100 i FIG. 8. Raster plot of the ﬁring activity in the quenched network with N = 100 000 and coupling J = 0.8. To illustrate the busting activity of single neurons, we display the spiking activity of only 100 neurons over a short time interval. -15 -10 -5 0 R<I> [mV] 0 20 40 60 80 <ν> [Hz] J=0.8 J=0.5 J=0.25 0 50 100 150 200 t [ms] 0 20 40 60 80 100 i FIG. 9. Firing rate ⟨ν⟩versus the synaptic “current” R⟨I⟩for different coupling strengths for the quenched network (solid line and red circles). Zero coupling corresponds to the rightmost point, where the synaptic current obviously vanishes. The vertical dashed line separates the region where neurons operate above (right) from below (left) threshold. The green crosses report the outcome of the annealed network. The two crosses deviating from the solid line belong to J = 0.5 and 0.8 in the annealed setup. FIG. 8. Raster plot of the ﬁring activity in the quenched network with N = 100 000 and coupling J = 0.8. To illustrate the busting activity of single neurons, we display the spiking activity of only 100 neurons over a short time interval. correlations built while integrating the LIF equation are not negligible [33]. A. Correlations between membrane potential and synaptic current The counterintuitive activity displayed by the quenched network requires an explanation. We have veriﬁed that the effective self-induced excitation is not the result of a sym- metry breaking: All neurons (both excitatory and inhibitory) behave in the same way, as they should. More instructive information can be extracted by exploring the correlations between the actual value of the membrane potential and the quality (excitatory versus inhibitory) of the spike received by a given neuron. In other words, we have computed the relative fraction sE(V )dV of excitatory spikes received when V ∈ [V,V + dV ]. If the receiving times were uncorrelated with the membrane potential, then sE(V ) would be independent of V and equal to b. Actually, this is expected within the framework of a δ-correlated input signal as assumed in Ref. [4]. In the asynchronous regime, the average input current induced by the synaptic coupling is ⟨I⟩= J R (nE −gnI) = KJ R ν[b(g + 1) −g], where nE (nI) denotes the number of excitatory (inhibitory) spikes received per time unit. Depending on whether R0⟨I⟩ is larger or smaller than Vth −RI0 = −4, the neuron operates either above or below threshold. In fact, in the latter case, the velocity ﬁeld crosses the zero axis below the threshold Vth, preventing threshold passing. In Fig. 9, we plot the ﬁring rate versus ⟨I⟩for different values of the coupling strength J (see the solid line): increasing J corresponds to moving leftward along the curve, starting from the rightmost point, which corresponds to the uncoupled limit. Upon increasing J, ⟨I⟩decreases monotonically: This is the consequence of the prevalent inhibition. At the same time, the ﬁring rate, after an initial drop, starts growing; this happens for J ≈0.25, as it can be inferred by comparing with Fig. 2(a). The increase continues also when the neuron operates below threshold and surpasses the activity of the uncoupled regime. where nE (nI) denotes the number of excitatory (inhibitory) spikes received per time unit. Depending on whether R0⟨I⟩ is larger or smaller than Vth −RI0 = −4, the neuron operates either above or below threshold. In fact, in the latter case, the velocity ﬁeld crosses the zero axis below the threshold Vth, preventing threshold passing. In Fig. B. Power spectrum Fourier analysis offers the opportunity for additional ver- iﬁcation of the validity of the RP approximation. In Fig. 4, we compare the power spectrum of the single-neuron activity after the ﬁrst iterate obtained under the RP approximation [31] (see the purple curve) with the spectrum exhibited by the quenched network. The agreement is quite good for all three tested coupling strengths, the major discrepancy being the absence of peaks at multiples of νd = 1818 Hz for J = 0.8, which cannot be reproduced by the RP approximation, since the delay is not included in such formulation. Next, we have directly estimated the correlations among the phases of the Fourier modes by computing D( f ) [see Eq. (3)] both for the quenched network and the RP approx- imation (see upper and lower curves in Fig. 6, respectively). Phase correlations appear to be small in both cases (look at the vertical scale): We attribute the larger amplitude exhibited by the quenched network to the presence of a residual collective dynamics, absent by deﬁnition in the RP approximation. A k+1 k We have implemented this approach with an Euler inte- gration step δt = 10−3 ms starting from the initial condition Pq, the best proxy for the asynchronous regime. The ﬁrst 33 iterates are reported in Fig. 5, where we see that they move away from Pr (see the blue squares) and approach a seemingly stable ﬁxed point Ps. The relatively large difference between Ps and Pq suggests that this approximation scheme is not as accurate as the RP method and implicitly means that the phase 023103-7 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) ULLNER, POLITI, AND TORCINI 0 50 100 150 200 t [ms] 0 20 40 60 80 100 i FIG. 8. Raster plot of the ﬁring activity in the quenched network with N = 100 000 and coupling J = 0.8. To illustrate the busting activity of single neurons, we display the spiking activity of only 100 neurons over a short time interval. correlations built while integrating the LIF equation are not negligible [33]. -15 -10 -5 0 R<I> [mV] 0 20 40 60 80 <ν> [Hz] J=0.8 J=0.5 J=0.25 FIG. 9. Firing rate ⟨ν⟩versus the synaptic “current” R⟨I⟩for different coupling strengths for the quenched network (solid line and red circles). Zero coupling corresponds to the rightmost point, where the synaptic current obviously vanishes. B. Synaptic current: An Ornstein-Uhlenbeck process The role of correlations can be analyzed from a different point of view: Since the neuron is typically under the action of a negative current, its membrane potential is kept away from threshold (V < Vth). Only when relatively strong pos- itive ﬂuctuations of the input current arise, can the neuron overcome the threshold and emit a spike. If the correlations are sufﬁciently long lasting, the ﬂuctuation may stand long enough to allow for the emission of a sequence of spikes and give rise to a “burst.” This mechanism has already been investigated in the past to quantify the spiking activity of a subthreshold neuron subject to Ornstein-Uhlenbeck (OU) noise, ﬁnding that a long correlation time gives rise to a bursting activity [30,34]. Unfortunately, we cannot make use of their formulas, since the correlation time is not much longer than τ. We can, nevertheless, proceed in a purely numerical way by approximating the input current I with the outcome of an OU stochastic equation, namely, FIG. 10. (a) Fraction sE of excitatory spikes versus the current value V for J = 0.8. The horizontal line corresponds to the average global fraction b = 0.8. (b) Effective velocity ﬁeld Feff as from Eq. (6). where we plot the effective velocity ﬁeld (6) Feff = RI0 −V + KJτ⟨ν⟩[sE(1 + g) −g], (6) τc ˙I = ⟨I⟩−I + ξ, where the V -dependent sE replaces b. Interestingly, the ef- fective velocity ﬁeld does not cross the zero axis below threshold, showing that the neuron effectively operates above threshold, in spite of R⟨I⟩< Vth −RI0. So, we can conclude that including the V dependence of sE into the neural dy- namics helps solve the paradox of a neuron operating on average below threshold. On a more quantitative level, we can interpret Eq. (6) as a deterministic evolution equation and thereby compute the ﬁring rate νa as the inverse of the time needed to reach the threshold Vth, while starting from Vr (augmented by the refractory time). By inserting ν and sE(V ) values obtained from the network simulations for J = 0.8, we ﬁnd νa = 77 Hz, to be compared with the observed rate ν = 50 Hz. The agreement is not as good as one might have hoped for, but it should also be noted that Eq. (6) does not account for the (strong) input ﬂuctuations! A. Correlations between membrane potential and synaptic current 9, we plot the ﬁring rate versus ⟨I⟩for different values of the coupling strength J (see the solid line): increasing J corresponds to moving leftward along the curve, starting from the rightmost point, which corresponds to the uncoupled limit. Upon increasing J, ⟨I⟩decreases monotonically: This is the consequence of the prevalent inhibition. At the same time, the ﬁring rate, after an initial drop, starts growing; this happens for J ≈0.25, as it can be inferred by comparing with Fig. 2(a). The increase continues also when the neuron operates below threshold and surpasses the activity of the uncoupled regime. Instead, in Fig. 10(a), we see sizable deviations, especially in the vicinity of Vth, where sE is signiﬁcantly larger than b, hinting at a higher excitation than a priori foreseeable. We have veriﬁed that, as expected, the average of sE—computed along the V axis and weighted according to the stationary distribution P(V )—is equal to b; see the horizontal line. A perhaps more enlightening representation of the role played by the V dependence of sE is proposed in Fig. 10(b), 023103-8 QUANTITATIVE AND QUALITATIVE ANALYSIS OF … PHYSICAL REVIEW RESEARCH 2, 023103 (2020) -300 -200 -100 0 Vth v [mV] 0.76 0.8 0.84 sE -50 -40 -30 -20 0 10 20 v [mV] 0 20 40 60 Fef [mV] (a) (b) FIG. 10. (a) Fraction sE of excitatory spikes versus the current value V for J = 0.8. The horizontal line corresponds to the average global fraction b = 0.8. (b) Effective velocity ﬁeld Feff as from Eq. (6). This is precisely what we see in Fig. 10, where one can notice that sE is larger than b close to threshold. Consistency then imposes that sE < b further away. This is precisely what we see in Fig. 10, where one can notice that sE is larger than b close to threshold. Consistency then imposes that sE < b further away. B. Synaptic current: An Ornstein-Uhlenbeck process where ⟨I⟩is the average current observed in the numerical simulations of the quenched network, while τc is the input correlation time and, ﬁnally, ξ is a δ-correlated white noise ⟨ξ(t′)ξ(t′ + t)⟩= σ 2δ(t). We have thereby tuned τc and σ 2 until the neuron activity is characterized by the expected ﬁring rate and the corresponding Cv. For J = 0.8, we have found τc ≈160 ms and σ 2 ≈0.062. As a bonus, the resulting ISI distribution turns out to be quite similar to the expected one, the major difference being the peak which, instead of being located in T = 1 ms [see the inset in Fig. 3(c)], occurs for T ≈4 ms. Altogether, one can nevertheless conclude that the OU approximation provides a reasonable description of the input current. p We have implemented the same procedure for J = 0.5: In spite of the similar bursting activity, the neuron operates on average above threshold and we have not found any way to parametrize the OU process so as to reproduce the observed activity. On the other hand, a good reproduction of the neural activity is found for J = 1 by assuming τc = 145 and σ 2 = 0.28. Two interesting comments are in order: (i) upon increasing the coupling, the correlation time does not increase—it seems that τC ≈140−160 is an intrinsic property of the network, and (ii) the noise amplitude increases by more than a factor of 4, while passing from J = 0.8 to J = 1 and this is the reason why the ﬁring rate is larger in the latter case, even though the neuron operates much more below threshold. The increase of the effective noise can be attributed to two fac- tors: A minor contribution comes from the increased coupling strength (from 0.8 to 1); a more relevant contribution is the increased ﬂuctuations of the single-neuron activity quantiﬁed by the Cv. Moreover, the V dependence of sE still needs to be un- derstood. Some light can be shed by arguing as follows. C. Synaptic current: Symbolic correlations We conclude this section by looking at correlations from a different point of view. As shown in Sec. IV, the output activity of the single neuron is well approximated by a RP, but we do not expect the same to be true for the input, obtained by superposing K independent such processes. To investi- gate the way correlations manifest themselves, we separately computed the ISI distribution of all excitatory and inhibitory spikes received by a given neuron. Altogether, this proves that the relevant correlations are contained in the symbolic representation of the spike se- quence. The very negative values of the membrane potential displayed in Fig. 12 can raise doubts about the validity of the LIF model. However, it should be recalled that the exponen- tial integrate and ﬁre neuron [35], successfully employed to reproduce the neurophysiological characteristics of pyramidal neurons in the cortex [36], reduces to the LIF neuron for very negative membrane potentials. Moreover, the strong coupling strength selected in this paper has been chosen to highlight the role of correlations, but the phenomenon persists at lower coupling strengths too. p y g In Fig. 11, we report the ISI distribution of excitatory spikes (inhibitory spikes follow the same statistics) for both the original quenched network (black curve) and within the RP approximation (red curve). T represents the average sep- aration between consecutive spikes, i.e., T is equal to the average single-neuron ISI divided by 800—the total number of incoming excitatory synaptic connections. The red curve follows a clean Poissonian distribution, while the quenched network exhibits a slower than exponential decay (in this time range); furthermore, in the latter case, the ﬁrst channel is very large because of the unavoidable presence of avalanches occurring in the quenched setup (see Ref. [6]). We attribute most of the deviations from a pure exponential to the residual presence of collective dynamics. In any case, this discrepancy is a minor issue: The relevant correlations are those between excitatory and inhibitory spikes, as revealed by the following test. We have fed a single neuron with two different signals: (i) a perfectly Poisson process composed of independent excitatory and inhibitory spikes and (ii) a synthetic signal built by assuming a Poisson distribution of consecutive spikes with the same rate as the quenched network, but keeping the original symbolic ordering (i.e., EEIEEEIE . . C. Synaptic current: Symbolic correlations ., where the letters E/I means that the spike is either excitatory or inhibitory) observed in the quenched network. B. Synaptic current: An Ornstein-Uhlenbeck process The horizontal axis is rescaled by the mean ﬁring rate of the respec- tive population of excitatory spikes. -200 -100 0 v [mV] 0 0.005 0.01 P(v) FIG. 12. Probability density of the membrane potential P(v). The black curve exhibiting the divergence in v = 10 corresponds to the synthetic signal described in text; the red curve is obtained by feeding the neuron with a purely Poisson process with the same ﬁring rate. -200 -100 0 v [mV] 0 0.005 0.01 P(v) FIG. 12. Probability density of the membrane potential P(v). The black curve exhibiting the divergence in v = 10 corresponds to the synthetic signal described in text; the red curve is obtained by feeding the neuron with a purely Poisson process with the same ﬁring rate. 0 2 4 6 10 12 14 T/T 10 -5 10 -4 10 -3 10 -2 10 -1 10 0 Q(T) -200 -100 0 v [mV] 0 0.005 0.01 P(v) FIG. 12. Probability density of the membrane potential P(v). The black curve exhibiting the divergence in v = 10 corresponds to the synthetic signal described in text; the red curve is obtained by feeding the neuron with a purely Poisson process with the same ﬁring rate. FIG. 11. The effective distribution ¯Q(T ) of interexcitatory spikes received by a single neuron for J = 0.8. The black curve refers to the quenched network, the red one to the renewal process approximation. The horizontal axis is rescaled by the mean ﬁring rate of the respec- tive population of excitatory spikes. the original distribution. On a quantitative level, the resulting ﬁring rate, the Cv, and the PDF of the ISIs are very close to the values exhibited by the RP approximation (deviations are smaller than 1%): see the inset of Fig. 3(c). On the other hand, the red curve, originating from the strictly Poisson process is shifted toward very negative v values and nearly vanishes close to the threshold, suggesting a very low ﬁring activity as indeed observed. B. Synaptic current: An Ornstein-Uhlenbeck process Let us introduce the joint probability P(E, L) that an excitatory spike reaches the neuron, when its membrane potential V ∈ L = [V0,Vth], where V0 is selected as the point where sE = b, The standard Bayesian inference rule implies that P(E\|L) = P(L\|E) P(L) P(E), where P(A\|B) denotes the probability of observing A, given B; moreover P(E) = b, while P(L) is the probability of V > V0, and P(E\|L) is just the average of sE over L. Let us now focus on P(L\|E)/P(L): This is the probability of V > V0 when an excitatory spike arrives (rescaled to the unconditional probability to stay in L). If excitatory spikes arrive in bursts, for many of them the corresponding V value is relatively large as a consequence of the excitation provided by the previous spikes. Therefore, it is natural to expect P(L\|E)/P(L) > 1. where P(A\|B) denotes the probability of observing A, given B; moreover P(E) = b, while P(L) is the probability of V > V0, and P(E\|L) is just the average of sE over L. Let us now focus on P(L\|E)/P(L): This is the probability of V > V0 when an excitatory spike arrives (rescaled to the unconditional probability to stay in L). If excitatory spikes arrive in bursts, for many of them the corresponding V value is relatively large as a consequence of the excitation provided by the previous spikes. Therefore, it is natural to expect P(L\|E)/P(L) > 1. 023103-9 PHYSICAL REVIEW RESEARCH 2, 023103 (2020) ULLNER, POLITI, AND TORCINI 0 2 4 6 10 12 14 T/T 10 -5 10 -4 10 -3 10 -2 10 -1 10 0 Q(T) FIG. 11. The effective distribution ¯Q(T ) of interexcitatory spikes received by a single neuron for J = 0.8. The black curve refers to the quenched network, the red one to the renewal process approximation. The horizontal axis is rescaled by the mean ﬁring rate of the respec- tive population of excitatory spikes. 0 2 4 6 10 12 14 T/T 10 -5 10 -4 10 -3 10 -2 10 -1 10 0 Q(T) FIG. 11. The effective distribution ¯Q(T ) of interexcitatory spikes received by a single neuron for J = 0.8. The black curve refers to the quenched network, the red one to the renewal process approximation. VI. CONCLUSIONS AND OPEN PROBLEMS In this paper, we have shown that upon increasing the coupling strength J (and for J > 0.25), a slightly inhibitory sparse network of LIF neurons operates increasingly below threshold and yet ﬁres at an increasingly fast rate. This claim is supported by careful numerical simulations, tailored so as to marginalize the effects of collective synchronization. This counterintuitive, self-sustained activity observed in quenched networks, disappears in annealed networks, i.e., in setups where the synaptic connections are continuously randomly reshufﬂed. In the latter case, the neural activity is both weaker and more homogeneous (for J = 0.8, the ﬁring rate drops by a factor of about 4). The difference between quenched and annealed setups is reminiscent of replica sym- metry breaking [16], but the anomaly of the phenomenon is mitigated by the observation that the quenched-network The resulting membrane-potential distributions of the neu- ron are reported in Fig. 12. The black curve, obtained by using the above-mentioned synthetic signal, is very similar to 023103-10 QUANTITATIVE AND QUALITATIVE ANALYSIS OF … PHYSICAL REVIEW RESEARCH 2, 023103 (2020) 0 0.2 0.4 0.6 0.8 J 0 20 40 <ν> [Hz] (a) 0 0.2 0.4 0.6 0.8 J 0 20 40 <ν> [Hz] 0 0.2 0.4 0.6 0.8 J 0 1 2 3 4 <Cv> (a) (b) FIG. 13. Neural activity for different delay times. The solid curves all refer to τd = 0.55, while crosses refer to τd = 0.3. All simulations are performed in a network with N = 105 neurons and a connectivity K = 1000. 0 0.2 0.4 0.6 0.8 J 0 1 2 3 4 <Cv> (b) different delay values conﬁrm a substantial independence of the outcome on τd. In Fig. 13, we compare the ﬁring rate and the Cv obtained for τd = 0.3 with the original simulations (performed for τd = 0.55). p Last but not least, we wish to comment on the peculiar behavior of the network observed for large coupling strengths. The strong ﬁring activity is self-sustained by its burstiness (signaled by the large Cv values), which, de facto, provides the relatively long correlations required to let a neuron below threshold ﬁre. This clariﬁes the reason why the theoretical formula based on the assumption of δ-correlated current ﬂuctuations fails to reproduce this regime. VI. CONCLUSIONS AND OPEN PROBLEMS Recently, a more sophisticated self-consistent approach has been developed, where the Fokker-Planck equation has been augmented to account for temporal correlations in the synaptic current [38]. Its (numerical) implementation to a weak-bursting regime looks promising. It will be worth exploring its validity in a more inhibition-dominated regime such as the one explored in this paper. Interestingly, the bursting activity is reproduced also assuming a strictly Poisson ISI distribution, but retaining the correlations contained in the symbolic representation of the spike types (i.e., excitatory versus inhibitory). A simple quantiﬁcation of such correlations might open yet another route for a quantitative characterization of the neural activity. FIG. 13. Neural activity for different delay times. The solid curves all refer to τd = 0.55, while crosses refer to τd = 0.3. All simulations are performed in a network with N = 105 neurons and a connectivity K = 1000. dynamics can be reproduced to a high degree of accuracy by an approach, the RP approximation, which does not take into account the structure of the synaptic connections. Still, the comparison between quenched and annealed dynamics (see Fig. 9 for the most enlightening representation) seems to suggest the presence of a phase transition when J is increased. It looks like the two regimes deviate from one another above J ≈0.25. This is reminiscent of the claim made by Ostojic about the existence of two distinct asynchronous regimes [11]. This claim has been criticised in Ref. [17]; we are also unable to ﬁnd evidence of a qualitative difference between the two regimes (above and below a supposedly critical point Jc). Finally, we wish to stress that no speciﬁc properties of the LIF neurons have been invoked for our approach to be valid. Accordingly, we expect the RP approximation to be applicable to different neural models such as the exponential and the quadratic integrate and ﬁre neurons, or conductance-based models. Within the RP approximation, the neural activity is fully characterized by the ISI distribution. In the limit of large coupling strengths, such a distribution exhibits a power-law decay, similar to what was found while studying the response of a single neuron to OU processes [30,34] and similar to experimental observations made in the sensorimotor cortex of rats performing behavioral tasks [37]. It should, however, be noticed that in our case, the scaling range is much smaller than in the experimental observations. [11] S. Ostojic, Nat. Neurosci. 17, 594 (2014). ACKNOWLEDGMENTS A.T. received ﬁnancial support by the Excellence Initia- tive I-Site Paris Seine (Grant No. ANR-16-IDEX-008), by the Labex MME-DII (Grant No ANR-11-LBX-0023-01) (to- gether with A.P. and E.U.), and by the ANR Project ER- MUNDY (Grant No ANR-18-CE37-0014), all part of the French program Investissements d’Avenir. The recursive process based on the RP approximation proves rather accurate in spite of not taking into account the delay. It is therefore natural to ask whether this is also true in the quenched network. Simulations performed for [11] S. Ostojic, Nat. Neurosci. 17, 594 (2014). [1] A. S. Ecker, P. Berens, G. A. Keliris, M. Bethge, N. K. Logothetis, and A. S. Tolias, Science 327, 584 (2010). [12] C. van Vreeswijk and H. Sompolinsky, Science 274, 1724 (1996). [2] A. Renart, J. de la Rocha, P. Bartho, L. Hollender, N. Parga, A. Reyes, and K. D. Harris, Science 327, 587 (2010). [13] M. Volgushev, I. Kudryashov, M. Chistiakova, M. Mukovski, J. Niesmann, and U. T. Eysel, J. Neurophysiol. 92, 212 (2004). [3] R. M. Capocelli and L. M. Ricciardi, Kybernetik 8, 214 (1971). [14] A. Manwani and C. Koch, Neural Comput. 13, 1 (2001). [4] N. Brunel, J. Comput. Neurosci. 8, 183 (2000). [5] E. Ullner, A. Politi, and A. Torcini, Chaos 28, 081106 (2018), [15] L. A. Gatys, A. S. Ecker, T. Tchumatchenko, and M. Bethge, Phys. Rev. E 91, 062707 (2015). [6] A. Politi, E. Ullner, and A. Torcini, Eur. Phys. J. Spec. Top. 227, 1185 (2018). [16] M. Mézard, G. Parisi, and M. Virasoro, Spin Glass Theory and Beyond: An Introduction to the Replica Method and its Applications (World Scientiﬁc Publishing Co. Inc., Singapore, 1987), Vol. 9. [7] M. J. E. Richardson and R. Swarbrick, Phys. Rev. Lett. 105, 178102 (2010). [8] S. Olmi, D. Angulo-Garcia, A. Imparato, and A. Torcini, Sci. Rep. 7, 1577 (2017). [17] R. Engelken, F. Farkhooi, D. Hansel, C. van Vreeswijk, and F. Wolf, F1000Research 5, 2043 (2016). [9] B. Dummer, S. Wieland, and B. Lindner, Frontiers Comput. Neurosci. 8, 104 (2014). 8] E. Ullner and A. Politi, Phys. Rev. X 6, 011015 (2016 [10] R. F. O. Pena, S. Vellmer, D. Bernardi, A. C. Roque, and B. Lindner, Frontiers Comput. Neurosci. 12, 9 (2018). [19] A. Politi, A. Pikovsky, and E. Ullner, Eur. Phys. J. Spec. Top. 226, 1791 (2017). 226, 1791 (2017). ACKNOWLEDGMENTS 023103-11 ULLNER, POLITI, AND TORCINI PHYSICAL REVIEW RESEARCH 2, 023103 (2020) [20] W. Gerstner, W. M. Kistler, R. Naud, and L. Paninski, Neuronal Dynamics: From Single Neurons to Networks and Models of Cognition (Cambridge University Press, Cambridge, 2014). Additionally, one expects the single steps of the recursive pro- cedure are affected by statistical ﬂuctuations: We have veriﬁed that the uncertainty of ⟨ν⟩is about 0.05, while that of Cv is approximately 0.005. Additionally, one expects the single steps of the recursive pro- cedure are affected by statistical ﬂuctuations: We have veriﬁed that the uncertainty of ⟨ν⟩is about 0.05, while that of Cv is approximately 0.005. [21] D. Golomb, Scholarpedia 2, 1347 (2007). [30] T. Schwalger and L. Schimansky-Geier, Phys. Rev. E 77, 031914 (2008). [22] M. Mattia and P. D. Giudice, Neural Comput. 12, 2305 (2000). [22] M. Mattia and P. D. Giudice, Neural Comput [23] R. Brette, M. Rudolph, T. Carnevale, M. Hines, D. Beeman, J. M. Bower, M. Diesmann, A. Morrison, P. H. Goodman, F. C. Harris, M. Zirpe, T. Natschläger, D. Pecevski, B. Ermentrout, M. Djurfeldt, A. Lansner, O. Rochel, T. Vieville, E. Muller, A. P. Davison, S. El Boustani, and A. Destexhe, J. Comput. Neurosci. 23, 349 (2007). [31] No appreciable differences can be noticed while referring to the following iterates. [32] Leaving aside the zero-frequency channel which contributes to the average and is treated differently. [33] The quantitative differences with the results for Ps reported in Ref. [10] are quite likely to be attributed to the lack of accuracy in the integration scheme employed therein. [24] One should also remember that because of the refractory period, T > τR. [25] S. Ostojic, J. Neurophysiol. 106, 361 (2011). [34] R. Moreno-Bote and N. Parga, Phys. Rev. Lett. 92, 028102 (2004). [26] Here and everywhere power spectra are represented, they are normalized in such a way that the total power is obtained by integrating over all positive frequencies: 0 ⩽f < +∞. [35] N. Fourcaud-Trocmé, D. Hansel, C. Van Vreeswijk, and N. Brunel, J. Neurosci. 23, 11628 (2003). [27] J. Friedrich and W. Kinzel, J. Comput. Neurosci. 27, 65 (2009). [36] L. Badel, S. Lefort, R. Brette, C. C. Petersen, W. Gerstner, and M. J. Richardson, J. Neurophysiol. 99, 656 (2008). [28] B. Lindner, Phys. Rev. E 73, 022901 (2006). [37] Y. Tsubo, Y. Isomura, and T. Fukai, PLoS Comput. Biol. 8, e1002461 (2012). ACKNOWLEDGMENTS [29] In ﬁnite networks, sample-to-sample ﬂuctuations are expected. Simulations of ﬁve different networks show that the standard deviation of ⟨ν⟩is σν ≈0.4, while that of Cv is σC = 0.01. [29] In ﬁnite networks, sample-to-sample ﬂuctuations are expected. Simulations of ﬁve different networks show that the standard deviation of ⟨ν⟩is σν ≈0.4, while that of Cv is σC = 0.01. [38] S. Vellmer and B. Lindner, Phys. Rev. Res. 1, 023024 (2019). 023103-12
https://openalex.org/W4388423097	https://www.spiedigitallibrary.org/journals/advanced-photonics/volume-5/issue-6/060501/Shining-the-shortest-flashes-of-light-on-the-secret-life/10.1117/1.AP.5.6.060501.pdf	English	null	Shining the shortest flashes of light on the secret life of electrons	Advanced photonics	2,023	cc-by	3,391	Shining the shortest flashes of light on the secret life of electrons Margarita Khokhlova Emilio Pisanty * and Amelle Zaïr Attosecond pulses are born in the extreme nonlinear interaction of high-intensity laser pulses with matter – conventionally with noble gases, but increasingly with solid-state media – in a process known as high-harmonic generation (HHG).8 HHG marks the growth of nonlinear optics from the finding of second-harmonic generation13 to the observation of nonperturbative processes with harmonic order that can now be measured in the thousands14 (Fig. 1). The first to cross the finish line was Pierre Agostini and his team, also at CEA in Saclay, who built a stable HHG setup and successfully timed the duration of the radiation bursts in the train down to a quarter of a femtosecond,9 i.e., 250. To do this, they used a fre- quency-domain technique now known as RABBITT33,34 (attosecond science acronyms have a distinct “animal” theme35–37), which looks at interference patterns in electrons ionized by the attosecond pulse train in the presence of a mid-intensity replica of the driving laser. The 2023 Nobel Prize in Physics awards the discovery of HHG by Anne L’Huillier and her team in 1988,8 and the subsequent use of this process by Pierre Agostini and Ferenc Krausz to experimentally demonstrate pulses of light shorter than one femtosecond.9,10 Almost simultaneously, Ferenc Krausz and his team at the Technical University of Vienna reported the generation of the first isolated HHG occurs when a strong and ultrafast laser drives the atoms in a gas, with enough intensity to go beyond the perturbative regime. The atoms can then up-convert the frequency of the laser, by combining large numbers of laser photons into single high-frequency harmonics. In contrast with “standard” harmonic generation, where the efficiency of the conversion drops exponentially with the number of laser photons being combined, L’Huillier and her team at CEA in Saclay, France, observed a long, flat plateau of harmonics (somewhat reminiscent of an optical frequency comb15,16) emitted with roughly equal intensities,8 which had first been glimpsed one year previously.17 Fig. 1 An artist’s idea of weak and strong fields interacting with matter: the weak field produces linear and low-order harmonic response to the laser field, while the strong one breaks the matter and generates high-order harmonics. Applying some imagination, one can even attribute the sound of breaking strings to attosecond pulses. Courtesy of Vasily Strelkov and Inna Midzyanovskaya. The process is best understood via the so-called three-step model, discovered independently by Paul Corkum and by Kenneth Kulander et al. Shining the shortest flashes of light on the secret life of electrons Margarita Khokhlova Emilio Pisanty * and Amelle Zaïr Margarita Khokhlova, Emilio Pisanty,* and Amelle Zaïr King’s College London, Department of Physics, London, United Kingdom Ever since the invention of laser pulses,1–5 one of the key directions for the development of laser technology has been the creation of ever shorter pulses of light. Over the past sixty years, improvements in tech- nology have pushed pulse durations down from nanoseconds through picoseconds to femtoseconds, giving us access to time-resolved studies of molecular nuclear motion and chemical reactions.6 Just in time for the 59th birthday of the standard (SI) prefixes “femto” and “atto,”7 the 2023 Nobel Prize in Physics was awarded to the latest landmark in this effort: the generation of attosecond pulses of light,8–10 which opens a window to the most fundamental timescale of the world around us – the timescale of electrons moving inside atoms, molecules, and con- densed matter. The bursts of radiation emitted by the recolliding electron occur on a subcycle timescale. It was realized early on after the discovery of the HHG plateau that, if the harmonics are locked in phase with each other, then their broad bandwidth would support subfemtosecond pulses.26–28 Moreover, the identification of the subcycle – attosecond – electron dynamics in the continuum that produces these flashes of light opened the door to the design of methods to confine the attosecond-pulse emis- sion to only one half-cycle of the driving laser, thus singling out one of the attosecond pulses in the train as an isolated attosecond pulse,29–31 a family of methods now known as “gating.”32 The scent of the attosecond world fired the starting pistol, and the race was now on: to generate attosecond pulses of light from HHG, either in a train or in isolation. This race presented significant chal- lenges, in the design and implementation of methods for temporal char- acterization which could reach such an unprecedented and demanding time resolution, and – for the generation of isolated pulses – the devel- opment of gating methods practical enough to be built in the lab. Attosecond pulses are the result of decades of combined advances in laser technology, raising the intensity boundary of laser pulses,11,12 and in nonlinear optics, which expanded to use these intense laser pulses to their full potential. Address all correspondence to Emilo Pisanty, emilio.pisanty@kcl.ac.uk © The Authors. Published by SPIE and CLP under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI. [DOI: 10.1117/1 .AP.5.6.060501] COMMENTARY COMMENTARY Shining the shortest flashes of light on the secret life of electrons Margarita Khokhlova Emilio Pisanty and Amelle Zaïr in 1993,18,19 building on previous work by Brunel20,21 and Kuchiev.22 The driving laser first rips an electron off an atom via tunnel ionization,23 then accelerates it in the continuum through the oscilla- tions of the electric field of the laser over one cycle, and finally clashes it with its parent ion at high speed, where it recombines with the hole it left behind and emits its considerable kinetic energy as a burst of light. This process then repeats every half-cycle of the driving laser, produc- ing a train of pulses. The simple classical picture was confirmed by experiments correlating HHG to recollision-induced double ioniza- tion,24 and it was shortly followed by an analytical quantum-mechanical theory developed by Maciej Lewenstein and co-workers.25 Fig. 1 An artist’s idea of weak and strong fields interacting with matter: the weak field produces linear and low-order harmonic response to the laser field, while the strong one breaks the matter and generates high-order harmonics. Applying some imagination, one can even attribute the sound of breaking strings to attosecond pulses. Courtesy of Vasily Strelkov and Inna Midzyanovskaya. 060501-1 Advanced Photonics Nov∕Dec 2023 • Vol. 5(6) COMMENTARY 4. F. J. McClung and R. W. Hellwarth, “Giant optical pulsations from ruby,” J. Appl. Phys. 33(3), 828 (1962). attosecond pulse.10 This was the result of a long development campaign to push the duration of strong laser pulses down to the few-cycle (and even single-cycle) regime. With such a short driver, the HHG emission is effectively confined to only a single burst, a technique now known as amplitude gating. Moreover, Krausz and his team managed to measure the duration of the emission down to 650 attoseconds, using a cross- correlation technique38 closely related to a now-standard method called the attosecond streak camera.39,40 5. J. M. Dudley, “Light, lasers, and the Nobel Prize,” Adv. Photonics 2(5), 050501 (2020). 6. A. Zewail, “Femtochemistry: atomic-scale dynamics of the chemi- cal bond using ultrafast lasers,” Nobel Lecture, 1999, https://www. nobelprize.org/prizes/chemistry/1999/zewail/lecture/ (accessed October 2023). 7. International Conference on Weights and Measures, Resolution 8, International System of Units, Proc. 12th CGPM, p. 94, (1964). In the two decades since these breakthroughs, the floodgates for discovery have opened wider and wider, allowing for a number of measurements and observations that were considered impossible or unthinkable for decades. Shining the shortest flashes of light on the secret life of electrons Margarita Khokhlova Emilio Pisanty * and Amelle Zaïr Attosecond pulses have been used to track nuclear motion,41 and later to observe faster and faster dynamics, in- cluding direct tracking of valence electron motion in atoms42 and bio- molecules,43 and of the even faster motion of core-shell electrons.44 They have also provided views of the time-resolved build-up of quan- tum interference patterns in atomic spectra,45 the interference of the various quasi-classical electron trajectories that produce HHG,46 and of the time-resolved coupling of electron and phonon degrees of freedom.47 8. M. Ferray et al., “Multiple-harmonic conversion of 1064 nm radi- ation in rare gases,” J. Phys. B: At. Mol. Opt. Phys. 21, L31 (1988). 9. P. M. Paul et al., “Observation of a train of attosecond pulses from high harmonic generation,” Science 292(5522), 1689 (2001). 10. M.Hentscheletal.,“Attosecondmetrology,”Nature414,509(2001). 11. G. Mourou, “Extreme light physics and application,” Nobel Lecture, 2018, https://www.nobelprize.org/prizes/physics/2018/ mourou/lecture/ (accessed October 2023). 12. D. Strickland, “Generating high-intensity ultrashort optical pulses,” Nobel Lecture, 2018, https://www.nobelprize.org/ prizes/physics/2018/strickland/lecture/ (accessed October 2023). 13. N. Bloembergen, “Nonlinear optics and spectroscopy,” Nobel Lecture, 1981, https://www.nobelprize.org/prizes/physics/1981/ bloembergen/lecture/ (accessed October 2023). One standout example is the application of the attosecond streak camera to directly observe the oscillations of the electric field of a light wave,48 providing a new and fresh answer to the question of optical coherence.49,50 This possibility breaks many of the assumptions held during the construction of quantum mechanics in the 1920s and 1930s, similarly to the prospects offered by the manipulation of individual quantum systems.51,52 g 14. T. Popmintchev et al., “Bright coherent ultrahigh harmonics in the keV X-ray regime from mid-infrared femtosecond lasers,” Science 336(6086), 1287 (2012). 15. J. L. Hall, “Defining and measuring optical frequencies: the optical clock opportunity – and more,” Nobel Lecture, 2005, https://www.nobelprize.org/prizes/physics/2005/hall/lecture/ (ac- cessed October 2023). The attosecond pulses produced via HHG have also been joined by pulses from additional sources, including other high-order parametric processes,53 oscillating relativistic plasma mirrors,54 and, most impor- tantly, facility-scale X-ray free-electron lasers.55 XFELs also offer at- tosecond capabilities, at high brightness, and with a nuanced set of trade-offs regarding coherence and timing precision. This wider set of sources promises to further enrich our ability to probe the microscopic world and its attosecond dynamics. 16. T. W. Hänsch, “Passion for precision,” Nobel Lecture, 2005, https://www.nobelprize.org/prizes/physics/2005/hansch/lecture/ (accessed October 2023). 17. A. McPherson et al., “Studies of multiphoton production of vacuum-ultraviolet radiation in the rare gases,” J. Opt. Soc. Am. B 4(4), 595 (1987). 18. Shining the shortest flashes of light on the secret life of electrons Margarita Khokhlova Emilio Pisanty * and Amelle Zaïr P. B. Corkum, “Plasma perspective on strong field multiphoton ionization,” Phys. Rev. Lett. 71(13), 1994 (1993). Looking forward, the future of attosecond science promises signifi- cant and inspiring innovations, both in the advance of attosecond sources and in their applications. On the side of the sources, it has recently become possible to use attosecond pulses to both pump and probe ultrafast phenomena in the same experiment,56 and attosecond interferometry has reached extreme levels of precision,57 both of which hold significant promise in reaching new regimes and dynamics. The optical control of attosecond pulses continues to increase, including the use of structured light,58 tailored polarizations,59 chiral states of light,60 attosecond frequency combs based on femtosecond enhancement cavities,61 and detailed control of the HHG wavefronts.62 19. K. C. Kulander, K. J. Schafer, and J. L. Krause, “Dynamics of short- pulse excitation, ionization and harmonic conversion,” in Super- Intense Laser Atom Physics, B. Piraux, A. L’Huillier, and K. Rzążewski, Eds., NATO Advanced Studies Institute Series B: Physics, 316, 95–110, Springer, Boston, Massachusetts (1993). y p g ( ) 20. F. Brunel, “Harmonic generation due to plasma effects in a gas undergoing multi-photon ionization in the high-intensity limit,” J. Opt. Soc. Am. B 7, 521 (1990). p 21. P. B. Corkum, N. H. Burnett, and F. Brunel, “Above-threshold ionization in the long-wavelength limit,” Phys. Rev. Lett. 62(11), 1259 (1989). Attosecond science also continues to expand the range of systems it can study, from HHG in solids63 to liquids64 and nanostructures.65 Its growth has also made it possible to build fertile interfaces with other branches of physics, including quantum optics,66 and quantum informa- tion processing,67 which hold substantial promise of innovation for this young – but now mature – and dynamic discipline. 22. M. Yu. Kuchiev, “Atomic antenna,” JETP Lett. 45(7), 404 (1987) [Pis’ma Zh. Eksp. Teor. Fiz. 45(7), 319 (1987)]. 23. S. V. Popruzhenko, “Keldysh theory of strong field ionization: history, applications, difficulties and perspectives,” J. Phys. B: At. Mol. Opt. Phys. 47(20), 204001 (2014). 24. P. Dietrich et al., “High-harmonic generation and correlated two- electron multiphoton ionization with elliptically polarized light,” Phys. Rev. A 50, R3585 (1994). References Goulielmakis et al., “Real-time observation of valence electron motion,” Nature 466, 739 (2010). 43. F. Calegari et al., “Ultrafast electron dynamics in phenylalanine initiated by attosecond pulses,” Science 346, 336 (2014). 44. T. Barillot et al., “Correlation-driven transient hole dynamics resolved in space and time in the isopropanol molecule,” Phys. Rev. X 11, 031048 (2021). 45. C. Ott et al., “Lorentz meets Fano in spectral line shapes: a uni- versal phase and its laser control,” Science 340, 716 (2013). Emilio Pisanty is a Royal Society University Research Fellow at King’s College London. He received his BSc degree from the National Autonomous University of Mexico (UNAM), and a PhD in Controlled Quantum Dynamics from Imperial College London. He has worked at the Max Born Institute in Berlin and in ICFO – The Institute of Photonic Sciences in Barcelona. His current research interests include the inter- sections of attosecond science with structured light, catastrophe optics, and quantum optics. 46. A. 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https://openalex.org/W2051056784	https://asp-eurasipjournals.springeropen.com/counter/pdf/10.1155/2008/147407	English	null	Downsampling Non-Uniformly Sampled Data	EURASIP Journal on Advances in Signal Processing	2,007	cc-by	8,141	Recommended by T.-H. Li Decimating a uniformly sampled signal a factor D involves low-pass antialias ﬁltering with normalized cutoﬀfrequency 1/D followed by picking out every Dth sample. Alternatively, decimation can be done in the frequency domain using the fast Fourier transform (FFT) algorithm, after zero-padding the signal and truncating the FFT. We outline three approaches to decimate non- uniformly sampled signals, which are all based on interpolation. The interpolation is done in diﬀerent domains, and the inter- sample behavior does not need to be known. The ﬁrst one interpolates the signal to a uniformly sampling, after which standard decimation can be applied. The second one interpolates a continuous-time convolution integral, that implements the antialias ﬁlter, after which every Dth sample can be picked out. The third frequency domain approach computes an approximate Fourier transform, after which truncation and IFFT give the desired result. Simulations indicate that the second approach is particularly useful. A thorough analysis is therefore performed for this case, using the assumption that the non-uniformly distributed sampling instants are generated by a stochastic process. Copyright © 2008 F. Eng and F. Gustafsson. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Hindawi Publishing Corporation EURASIP Journal on Advances in Signal Processing Volume 2008, Article ID 147407, 10 pages doi:10.1155/2008/147407 Hindawi Publishing Corporation EURASIP Journal on Advances in Signal Processing Volume 2008, Article ID 147407, 10 pages doi:10.1155/2008/147407 Problem 1. The following is given: The sampling is determined by a nuisance event process. One typical example is data traﬃc in the Internet, where packet arrivals determine the sampling times and the queue length is the signal to be analyzed. Financial data, where the stock market valuations are determined by each transaction, is an- other example. (a) a sequence of non-uniform sampling times, tm, m = 1, ..., M; (a) a sequence of non-uniform sampling times, tm, m = 1, ..., M; (b) corresponding signal samples, u(tm); (c) a ﬁlter impulse response, h(t); and (c) a ﬁlter impulse response, h(t); and (d) a resampling frequency, 1/T. (d) a resampling frequency, 1/T. 1. INTRODUCTION A number of other applications and relevant references can be found in, for example, [1]. Downsampling is here considered for a non-uniformly sam- pled signal. Non-uniform sampling appears in many applica- tions, while the cause for nonlinear sampling can be classiﬁed into one of the following two categories. It should be obvious from the examples above that for most applications, the original non-uniformly sampled sig- nal is sampled much too fast, and that oscillation modes and interesting frequency modes are found at quite low frequen- cies compared to the inverse mean sampling interval. The problem at hand is stated as follows. The problem at hand is stated as follows. Frida Eng and Fredrik Gustafsson Department of Electrical Engineering, Link¨opings Universitet, 58183 Link¨oping, Sweden Correspondence should be addressed to Fredrik Gustafsson, fredrik@isy.liu.se Received 14 February 2007; Accepted 17 July 2007 Received 14 February 2007; Accepted 17 July 2007 Recommended by T.-H. Li Uniform sampling in secondary domain (a) First, if T/Tu = D is an integer, then (i) u(mTu) is ﬁl- tered giving z(mTu), and (ii) z(nT) = z(nDTu) gives the decimated signal. (b) Further, if T/Tu = R/S is a rational number, then a frequency domain method is known. It is based on (i) zero padding u(mTu) to length RM, (ii) computing the discrete Fourier transform (DFT), (iii) truncating the DFT a factor S, and ﬁnally computing the inverse DFT (IDFT), where the (I)FFT algorithm is used for the (I)DFT calculation. Conversion between arbitrary sampling rates has also been discussed in many contexts. The issues with eﬃcient implementation of the algorithms are investigated in [3– 6], and some of the results are beneﬁcial also for the non- uniform case. A major contribution in this work is a detailed analysis of the algorithms, where we assume additive random sampling, (ARS), tm = tm−1 + τm, (4) (4) Resampling and reconstruction are closely connected, since a reconstructed signal can be used to sample at desired time points. The task of reconstruction is well investigated for diﬀerent setups of non-uniform sampling. A number of iterative solutions have been proposed, for example, [1, 7, 8], several more are also discussed in [9]. The algorithms are not well-suited for real-time implementations and are based on diﬀerent assumptions on the sampling times, tm, such as bounds on the maximum separation or deviation from the nominal value mTu. where τm is stochastic additive sampling noise given by the known probability density function pτ(t). The theoretical re- sults show that the downsampled signal is unbiased under fairly general conditions and present an equivalent ﬁlter that generates z(t) = h ⋆u(t), where h depends on the designed ﬁlter h and the characteristic function of the stochastic dis- tribution. The paper is organized as follows. The algorithms are de- scribed in Section 2. The convolutional interpolation gives promising results in the simulations in Section 3, and the last sections are dedicated to this algorithm. In Section 4, theo- retic analysis of both ﬁnite time and asymptotic performance is done. The section also includes illustrative examples of the theory. Section 5 investigates an application example and is- sues with choosing the ﬁlter h(t), while Section 6 concludes the paper. Russel [9] also investigates both uniform and non- uniform resampling thoroughly. Uniform sampling in secondary domain Also, the desired intersampling time, T, is much larger than the original mean intersampling time, Some angular speed sensors give a pulse each time the shaft has passed a certain angle, so the sampling times depend on angular speed. Also biological signals such as ECGs are natu- rally sampled in the time domain, but preferably analyzed in another domain (heart rate domain). μT ≜E tm −tm−1 ≈tM M = Tu. (1) (1) 2 EURASIP Journal on Advances in Signal Processing Let ⌊x⌋denote the largest integer smaller than or equal to x. Find Reconstruction has long been an interesting topic in im- age processing, especially in medical imaging, see, for exam- ple, [16], where, in particular, problems with motion arti- facts are addressed. Arbitrary sampling distributions are al- lowed, and the reconstruction is done through resampling to a uniform grid. The missing pixel problem is given at- tention in [9, 17]. In [18], approximation of a function with bounded variation, with a band-limited function, is consid- ered and the approximation error is derived. Pointwise re- construction is investigated in [19], and these results will be used in Section 5. z(nT), n = 1, ..., N, N = tM T ≜M D , (2) (2) such that z(nT) approximates z(nT), where z(t) = h ⋆u(t) = h(t −τ)u(τ)dτ (3) (3) Here, we neither put any constraints on the non-uniform sampling times, nor assumptions on the signal’s function class. Instead, we take a more application-oriented approach, and aim at good, implementable, resampling procedures. We will consider three diﬀerent methods for converting from non-uniform to uniform sampling. The ﬁrst and third al- gorithm are rather trivial modiﬁcations of the time and frequency-domain methods for uniformly sampled data, re- spectively, while the second one is a new truly non-uniform algorithm. We will compare performance of these three. In all three cases, diﬀerent kinds of interpolation are possible, but we will focus on zero-order hold (nearest neighbor) and ﬁrst order hold (linear interpolation). Of course, which in- terpolation is best depends on the signal and in particular on its inter-sample behavior. Though we prefer to talk about decimation, we want to point out that the theories hold for any type of ﬁlter h(t). is given by convolution of the continuous-time ﬁlter h(t) and signal u(t). For the case of uniform sampling, tm = mTu, two well- known solutions exist; see, for example, [2]. Uniform sampling in secondary domain Russell argues against the iterative solutions, since they are based on analysis with ideal ﬁlters, and no guarantees can be given for approximate so- lutions. A noniternative approach is given, which assumes periodic time grids, that is, the non-uniformity is repeated. Another overview of techniques for non-uniform sampling is given in [10], where, for example, Ferreira [11] studies the special case of recovery of missing data and Lacaze [12] re- constructs stationary processes. 2.3. Interpolation in the frequency domain LP-ﬁltering is given by a multiplication in the frequency do- main, and we can form the approximate Fourier transform (AFT), [22], given by Riemann integration of the Fourier transform, to get Algorithm 3. This is also a known approach in the uniform sampling case, where the DFT is used in each step. The AFT is formed for 2N frequencies to avoid circular convolution. This corresponds to zero-padding for uniform sampling. Then the inverse DFT gives the estimate. The computation burden in the ﬁrst case is a limiting fac- tor in applications, and for the other two examples, the inter- polation is not exact. We consider a simple spline interpola- tion, followed by ﬁltering and decimation as in Algorithm 1. This is a slight modiﬁcation of the known solution in the uni- form case as was mentioned in Section 1. Algorithm 1 is optimal only in the unrealistic case where the underlying signal u(t) is piecewise constant between the samples. The error will depend on the relation between the original and the wanted sampling; the larger the ratio M/N, the smaller the error. If one assumes a band-limited signal, where all energy of the Fourier transform U( f ) is restricted to f < 0.5N/tM, then a perfect reconstruction would be pos- sible, after which any type of ﬁltering and sampling can be performed without error. However, this is not a feasible so- lution in practice, and the band-limited assumption is not satisﬁed for real signals when the sensor is aﬀected by addi- tive noise. Remark 3. The AFT used in Algorithm 3 is based on Rie- mann integration of the Fourier transform of u(t), and would be exact whenever u(t)e−i2π f t is constant between sampling times, which of course is rarely the case. As for the two previous algorithms, the approximation is less grave for large enough M/N. This paper does not include an investiga- tion of error bounds. More investigations of the AFT were done in [22]. 2.4. Complexity Remark 1. Algorithm 1 ﬁnds u(jTu) by zero-order hold in- terpolation, where of course linear interpolation or higher- order splines could be used. However, simulations not in- cluded showed that this choice does not signiﬁcantly aﬀect the performance. In applications, implementation complexity is often an issue. We calculate the number of operations, Nop, in terms of addi- tions (a), multiplications (m), and exponentials (e). As stated before, we have M measurements at non-uniform times, and want the signal value at N time points, equally spaced with T. 2.1. Interpolation in time domain This algorithm can be further analyzed using the inverse Fourier transform, and the results in [22], which will be done in Section 4.1. It is well described in literature how to interpolate a signal or function in, for instance, the following cases. Remark 2. Higher-order interpolations of (5) were studied in [23] without ﬁnding any beneﬁts. Remark 2. Higher-order interpolations of (5) were studied in [23] without ﬁnding any beneﬁts. (i) The signal is band-limited, in which case the sinc in- terpolation kernel gives a reconstruction with no error [20]. When the ﬁlter h(t) is causal, the summation is only taken over m such that tm < kT, and thus Algorithm 2 is ready for online use. (ii) The signal has vanishing derivatives of order n + 1 and higher, in which case spline interpolation of order n is optimal [21]. (iii) The signal has a bounded second-order derivative, in which case the Epanechnikov kernel is the optimal in- terpolation kernel [19]. 2. INTERPOLATION ALGORITHMS Reconstruction of functions with a convolutional ap- proach was done by [13], and later also by [14]. The sampling is done via basis functions, and reduces to the regular case if delta functions are used. These works are based on sampling sets that fulﬁll the non-uniform sampling theorem given in [15]. Time-domain interpolation can be used with subsequent ﬁl- tering. Since LP-ﬁltering is desired, we also propose two other methods that include the ﬁlter action directly. The main idea is to perform the interpolation at diﬀerent levels and the problem was stated in Problem 1. 3 3 F. Eng and F. Gustafsson For Problem 1, with Tu = tM/M, compute (1) t j m = arg min tm<jTu\|jTu −tm\|, (2) u(jTu) = u(t j m), (3) z(kT) = M j=1hd(kT −jTu)u(jTu), where hd(t) is a discrete time realization of the impulse response h(t). Algorithm 1: Time-domain interpolation. For Problem 1, compute (1) z(kT) = M m=1τmh(kT −tm)u(tm). Algorithm 2: Convolution interpolation. For Problem 1, with Tu = tM/M, compute (1) t j m = arg min tm<jTu\|jTu −tm\|, (2) u(jTu) = u(t j m), (3) z(kT) = M j=1hd(kT −jTu)u(jTu), For Problem 1, with Tu = tM/M, compute (1) t j m = arg min tm<jTu\|jTu −tm\|, (2) u(jTu) = u(t j m), (3) z(kT) = M j=1hd(kT −jTu)u(jTu), Algorithm 2: Convolution interpolation. where hd(t) is a discrete time realization of the impulse response h(t). where hd(t) is a discrete time realization of the impulse response h(t). and using Riemann integration, we get Algorithm 2. The al- gorithm will be exact if the integrand, h(kT −τ)u(τ), is con- stant between the sampling points, tm, for all kT. As stated before, the error, when this is not the case, decreases when the ratio M/N increases. Algorithm 1: Time-domain interpolation. 2.2. Interpolation in the convolution integral (i) Step (3) in Algorithm 1 is a linear ﬁlter, with one addi- tion and one multiplication in each term, Filtering of the continuous-time signal, u, yields z(kT) = h(kT −τ)u(τ)dτ, (5) N1 op = (1m + 1a)MN. (6) (6) EURASIP Journal on Advances in Signal Processing 4 The desired uniform sampling is given by the intersampling time T = 4 seconds. The non-uniform sampling is deﬁned by For Problem 1, compute (1) fn = n/2NT, n = 0,... ,2N −1, (2) U( fn) = M m=1τmu(tm)e−i2π fntm,n = 0,... ,N, (3) Z( fn) = Z( f2N−n)′ = H( fn) U( fn), n = 0,... ,N, (4) z(kT) = 1/2NT 2N−1 n=0 Z( fn)ei2πkT fn k = 0,... ,N −1. Here, Z′ is the complex conjugate of Z. Algorithm 3: Frequency-domain interpolation. tm = tm−1 + τm, (11) τm ∈Re(tl, th), (12) (11) ( ) (11) (11) (12) (12) and the limits tl and th are varied. In the simulation, N is set to 64 and the number of non-uniform samples are chosen so that tM > NT is assured. This is not in exact correspondence with the problem formulation, but assures that the results for diﬀerent τm-distributions are comparable. Algorithm 3: Frequency-domain interpolation. The samples are corrupted by additive measurement noise, Computing the convolution in step (3) in the fre- quency domain would require the order of M log 2(M) operations. u tm = s tm + e tm , (13) (13) where e(tm) ∈N(0, σ2), σ2 = 0.1. where e(tm) ∈N(0, σ2), σ2 = 0.1. p (ii) Algorithm 2 is similar to Algorithm 1, p (ii) Algorithm 2 is similar to Algorithm 1, The ﬁlter is a second-order LP-ﬁlter of Butterworth type with cutoﬀfrequency 1/2T, that is, The ﬁlter is a second-order LP-ﬁlter of Butterworth type with cutoﬀfrequency 1/2T, that is, N2 op = (2m + 1a)MN, (7) (7) h(t) = √ 2 π T e−(π/T √2)t sin π T √2t , t > 0, (14) H(s) = (π/T)2 s2 + √2π/Ts + (π/T)2 . (15) (14) where the extra multiplication comes from the factor τm. (iii) Algorithm 3 performs an AFT in step (2), frequency- domain ﬁltering in step (3) and an IDFT in step (4), (15) This setup is used for 500 diﬀerent realizations of fj, τm, and e(tm). This setup is used for 500 diﬀerent realizations of fj, τm, and e(tm). 2.2. Interpolation in the convolution integral N3op = (2m + le + la)2M(N + 1) + (lm)(N + 1) + (le + lm + la)2N2. (8) (8) We will test four diﬀerent rectangular distributions (12): τm ∈Re(0.1, 0.3), μT = 0.2, σT = 0.06, (16a) τm ∈Re(0.3, 0.5), μT = 0.4, σT = 0.06, (16b) τm ∈Re(0.4, 0.6), μT = 0.5, σT = 0.06, (16c) τm ∈Re(0.2, 0.6), μT = 0.4, σT = 0.12, (16d) Using the IFFT algorithm in step (4) would give Nlog2(2N) instead, but the major part is still MN. All three algorithms are thus of the order MN, though Algo- rithms 1 and 2 have smaller constants. All three algorithms are thus of the order MN, though Algo- rithms 1 and 2 have smaller constants. Studying work on eﬃcient implementation, for example, [9], performance improvements, could be made also here, mainly for Algorithms 1 and 2, where the setup is similar. and the mean values, μT, and standard deviations, σT, are shown for reference. For every run, we use the algorithms presented in the previous section and compare their results to the exact, continuous-time, result, y g Taking the length of the ﬁlter h(t) into account can sig- niﬁcantly improve the implementation speed. If the impulse response is short, the number of terms in the sums in Algo- rithms 1 and 2 will be reduced, as well as the number of extra frequencies needed in Algorithm 3. z(kT) = h(kT −τ)s(τ)dτ. (17) (17) We calculate the root mean square error, RMSE, We calculate the root mean square error, RMSE, 3. NUMERIC EVALUATION λ ≜ 1 N k z(kT) −z(kT) 2. (18) (18) We will use the following example to test the performance of these algorithms. The signal consists of three frequencies that are drawn randomly for each test run. The algorithms are ordered according to lowest RMSE, (18), and Table 1 presents the result. The number of ﬁrst, second and third positions for each algorithm during the 500 runs, are also presented. Figure 1 presents one example of the re- sult, though the algorithms are hard to be separated by visual inspection. The algorithms are ordered according to lowest RMSE, (18), and Table 1 presents the result. The number of ﬁrst, second and third positions for each algorithm during the 500 runs, are also presented. Figure 1 presents one example of the re- sult, though the algorithms are hard to be separated by visual inspection. Example 1. A signal with three frequencies, fj, drawn from a rectangular distribution, Re, is simulated s(t) = sin 2π f1t −1 + sin 2π f2t −1 + sin 2π f3t , (9) fj ∈Re 0.01, 1 2T , j = 1, 2, 3. (10) A number of conclusions can be drawn from the previous example. (10) 5 F. Eng and F. Gustafsson (iv) Algorithms 1 and 2 have similar RMSE statistics, though, of the two, Algorithm 2 performs slightly bet- ter in the mean, in all the four tested cases. (iv) Algorithms 1 and 2 have similar RMSE statistics, though, of the two, Algorithm 2 performs slightly bet- ter in the mean, in all the four tested cases. Table 1: RMSE values, λ in (18), for estimation of z(kT), in Example 1. The number of runs, where respective algorithm ﬁn- ished 1st, 2nd, and 3rd, is also shown. E[λ] Std(λ) 1st 2nd 3rd Setup in (16a) Alg. 1 0.281 0.012 98 258 144 Alg. 2 0.278 0.012 254 195 51 Alg. 3 0.311 0.061 148 47 305 Setup in (16b) Alg. 1 0.338 0.017 9 134 357 Alg. 2 0.325 0.015 175 277 48 Alg. 3 0.330 0.038 316 89 95 Setup in (16c) Alg. 1 0.360 0.018 6 82 412 Alg. 2 0.342 0.015 144 329 27 Alg. 3 0.341 0.032 350 89 61 Setup in (16d) Alg. 1 0.337 0.015 59 133 308 Alg. 2 0.331 0.015 117 285 98 Alg. 4. THEORETIC ANALYSIS Given the results for Algorithm 2 in the previous section, we will continue with a theoretic discussion of its behavior. We consider both ﬁnite time and asymptotic results. A small note is done on similar results for Algorithm 3. 200 190 180 170 160 150 140 130 120 110 100 Time, t −3 −2 −1 0 1 2 3 Figure 1: The result for the four algorithms, in Example 1, and a certain realization of (16c). The dots represent u(tm), and z(kT) is shown as a line, while the estimates z(kT) are marked with a ∗(Alg. 1), ◦(Alg. 2) and + (Alg. 3), respectively. 200 190 180 170 160 150 140 130 120 110 100 Time, t −3 −2 −1 0 1 2 3 3. NUMERIC EVALUATION 3 0.329 0.031 324 82 94 In this test, we ﬁnd that Algorithm 3 is most often number one, but Algorithm 2 is almost as good and more stable in its performance. It seems that the realization of the frequencies, fj, is not as crucial for the performance of Algorithm 2. As stated before, the performance also depends on the down- sampling factor for all the algorithms. The algorithms are comparable in performance and com- plexity. In the following, we focus on Algorithm 2, because of its nice analytical properties, its online compatibility, and, of course, its slightly better performance results. 4.1. Analysis of Algorithm 2 h(t) = F −1 H ⋆W( f ) (t), (26b) (26b) with W( f ) as in (20). p k 1 These known properties of W( f ) make it possible to ﬁnd E[z(kT)] and Var(z(kT)) for any given characteristic func- tion, ϕτ( f ), of the sampling noise, τk. ( f ) ( ) Furthermore, if the ﬁlter h(t) and signal u(t) belong to the Schwartz class, then S [24], f Furthermore, if the ﬁlter h(t) and signal u(t) belong to the Schwartz class, then S [24], Ez(kT) −→z(kT) if M m=1 pm(t) −→1 μT , M −→∞, (26c) Ez(kT) = z(kT) if M m=1 pm(t) = 1 μT , ∀M, (26d) τ The following lemma will be useful. (26c) Lemma 1 (see [22, Lemma 1]). Assume that the continuous- time function h(t) with FT H( f ) fulﬁlls the following condi- tions. (1) h(t) and H( f ) belong to the Schwartz class, S.1 with μT = E[τm], and pm(t) is the pdf for time tm. with μT = E[τm], and pm(t) is the pdf for time tm. with μT = E[τm], and pm(t) is the pdf for time tm. f g (2) The sum gM(t) = M m=1pm(t) obeys (2) The sum gM(t) = M m=1pm(t) obeys Proof. First of all, (5) gives Proof. First of all, (5) gives lim M−→∞ gM(t)h(t)dt = 1 μT h(t)dt = 1 μT H(0), (23) (23) (23) z(kT) = H(ψ)U(ψ)ei2πψkTdψ, (27a) (27a) for this h(t). f (3) The initial value is zero, h(0) = 0. f (3) The initial value is zero, h(0) = 0. z(kT) = U(ψ)ei2πψkT H(η)W(ψ −η)dη ≜H(ψ) dψ (27b) Then, it holds that Then, it holds that (27b) lim M−→∞ 1 −ϕτ( f )M 1 −ϕτ( f ) H( f )df = 1 μT H(0). (24) (24) which implies that we can identify H( f ) as the ﬁlter opera- tion on the continuous-time signal u(t), and (26a) follows. From Lemma 1 and (22), we get which implies that we can identify H( f ) as the ﬁlter opera- tion on the continuous-time signal u(t), and (26a) follows. which implies that we can identify H( f ) as the ﬁlter opera- tion on the continuous-time signal u(t), and (26a) follows. Proof. 4.1. Analysis of Algorithm 2 The proof is conducted using distributions from func- tional analysis and we refer to [22] for details. g From Lemma 1 and (22), we get From Lemma 1 and (22), we get E W( f ) y( f )df E W( f ) y( f )df = E τe−i2π f τ1 −ϕτ( f )M 1 −ϕτ( f ) y( f )df −→y(0) (28) Let us study the conditions on h(t) and H( f ) given in Lemma 1 a bit more. The restrictions from the Schwartz class could aﬀect the usability of the lemma. However, all smooth functions with compact support (and their Fourier trans- forms) are in S, which should suﬃce for most cases. It is not intuitively clear how hard (23) is. Note that, for any ARS case with continuous sampling noise distribution, pm(t) is approximately a Gaussian for higher m, and we can conﬁrm that, for a large enough ﬁxed t, (28) for any function y( f ) fulﬁlling the properties of Lemma 1. This gives E z(kT) = H(η)U(ψ)ei2πψkT E W(ψ −η) dψ dη −→ H(ψ)U(ψ)ei2πψkT dψ = z(kT), gM(t) = M m=1 1 √2πmσT e−(t−mμT)2/2mσ2 T −→1 μT , M −→∞, (25) (29) (29) (25) (25) when H( f ) and U( f ) behave as requested, and (26c) follows. Using the same technique as in the proof of Lemma 1, (26d) also follows. with μT and σT being the mean and the standard deviation of the sampling noise τ, respectively. The integral in (23) can then serve as some kind of mean value approximation, and the edges of gN(t) will not be crucial. Also, condition 3 fur- ther restricts the behavior of h(t) for small t, which will make condition 2 easier to fulﬁll. From the investigations in [22], it is clear that H( f ), in (27b), is the AFT of the sequence h(tm) (cf. the AFT of u(tm) in step (2) of Algorithm 3). Requiring that both h(t) and u(t) be in the Schwartz class is not, as indicated before, a major restriction. Though, some thought needs to be done for each speciﬁc case before apply- ing the theorem. Theorem 1. The estimate given by Algorithm 2 can be written Theorem 1. The estimate given by Algorithm 2 can be written as as g Algorithm 3 can be investigated analogously. 1 1h ∈s ⇔tkh(1)(t) is bounded, that is, h(1)(t) = θ(\|t\|−k), for all k, l ≥0. 4.1. Analysis of Algorithm 2 Here, we study the a priori stochastic properties of the es- timate, z(kT), given by Algorithm 2. For the analytical cal- culations, we assume that the convolution is symmetric, and get z(kT) = M m=1 τmh tm u kT −tm = M m=1 τm H(η)ei2πηtmdη U(ψ)ei2πψ(kT−tm)dψ = H(η)U(ψ)ei2πψkT M m=1 τme−i2π(ψ−η)tmdψ dη = H(η)U(ψ)ei2πψkTW ψ −η; tM 1 dψ dη Time, t Figure 1: The result for the four algorithms, in Example 1, and a certain realization of (16c). The dots represent u(tm), and z(kT) is shown as a line, while the estimates z(kT) are marked with a ∗(Alg. 1), ◦(Alg. 2) and + (Alg. 3), respectively. (19) with W f ; tM 1 = M m=1 τme−i2π f tm. (20) (i) Comparing a given algorithm for diﬀerent non- uniform sampling time pdf, Table 1 shows that pτ(t), in (16a), (16b), (16c), (16d), has a clear eﬀect on the performance. (20) Let (ii) Comparing the algorithms for a given sampling time distribution shows that the lowest mean RMSE is no guarantee of best performance at all runs. Algorithm 2 has the lowest E[λ] for setup (16a), but still performs worst in 10% of the cases, and for (16d), Algorithm 3 is number 3 in 20% of the runs, while it has the lowest mean RMSE. ϕτ( f ) = E e−i2π f τ = e−i2π f τ pτ(τ)dτ = F pτ(t) ( (21) (21) denote the characteristic function for the sampling noise τ. Here, F is the Fourier transform operator. Then, [22, Theo- rem 2] gives (iii) Usually, Algorithm 3 has the lowest RMSE (1st posi- tion), but the spread is more than twice as large (stan- dard deviation of λ), compared to the other two algo- rithms. E W( f ) = −1 2πi dϕτ( f ) df 1 −ϕτ( f )M 1 −ϕτ( f ) , (22) (22) EURASIP Journal on Advances in Signal Processing 6 where h(t) is given by where h(t) is given by where h(t) is given by where also an expression for the covariance, Cov(W( f )), is given. The expressions are given by straightforward calcula- tions using the fact that the sampling noise sequences τm are independent stochastic variables and tm = m k=1τk in (20). 4.2. Illustration of Theorem 1 Theorem 1 shows that the originally designed ﬁlter H( f ) is eﬀectively replaced by another linear ﬁlter H( f ) when using Algorithm 2. Since H( f ) only depends on H( f ) and the re- alization of the sampling times tm, we here study H( f ) to exclude the eﬀects of the signal, u(t), on the estimate. g First, we illustrate (26b) by showing the inﬂuence of the sampling times, or, the distribution of τm, on E[ H]. We use the four diﬀerent sampling noise distributions in (16a), (16b), (16c), (16d), using the original ﬁlter h(t) from (14) with T = 4 seconds. Figure 2 shows the diﬀerent ﬁlters H( f ), when the sampling noise distribution is changed. We con- clude that both the mean and the variance aﬀect \|E[ H( f )]\|, and that it seems possible to mitigate the static gain oﬀset from H( f ) by multiplying z(kT) with a constant depending on the ﬁlter h(t) and the sampling time distribution pτ(t). Figure 3: An example of the ﬁlter h(tm) given by (31), (16b), and T = 4. Second, we show that E H −→H when M increases, for a smooth ﬁlter with compact support, (26c). Here, we use a close ﬁt also at higher frequencies. The convergence of the magnitude of the ﬁlter is clearly shown in Figure 4. h(t) = 1 4d f cos π 2 t −d f T d f T 2 , t −d f T < d f T, (31) 4.1. Analysis of Algorithm 2 z(kT) = h ⋆u(kT), (26a) (26a) Theorem 2. The estimate given by Algorithm 3 can be written as as z(kT) = h ⋆u(kT), (30a) z(kT) = h ⋆u(kT), (30a) 7 7 F. Eng and F. Gustafsson 10−1 10−2 Frequency, f (Hz) 0.85 0.88 0.91 0.94 0.97 1 Re(0.1, 0.3) Re(0.3, 0.5) Re(0.4, 0.6) Re(0.2, 0.6) H( f ) Figure 2: The true H( f ) (thick line) compared to \|E[H( f )]\| given by (14) and Theorem 1, for the diﬀerent sampling noise distribu- tions in (16a), (16b), (16c), (16d), and M = 250. where h(t) is given by the inverse Fourier transform of H( f ) = 1 2NT N−1 n=0 H fn e−i2π( f −fn)kTW( fn −f ) + 2N−1 n=N H −fn e−i2π( f −fn)kTW −fn −f , (30b) and W( f ) was given in (20). and W( f ) was given in (20). and W( f ) was given in (20). Proof. First, observe that real signals u(t) and h(t) give U( f )′ = U(−f ) and H( f )′ = H(−f )′, respectively, the rest is completely in analogue with the proof of Theorem 1, with one of the integrals replaced with the corresponding sum. Numerically, it is possible to conﬁrm that the require- ments on pm(t), in (26c), are true for Additive Random Sam- pling, since pm(t) then converges to a Gaussian distribution with μ = mμT and σ2 = mσT. A smooth ﬁlter with compact support is noncausal, but with ﬁnite impulse response (see e.g., the optimal ﬁlter discussed in Section 5). Figure 2: The true H( f ) (thick line) compared to \|E[H( f )]\| given by (14) and Theorem 1, for the diﬀerent sampling noise distribu- tions in (16a), (16b), (16c), (16d), and M = 250. A noncausal ﬁlter is desired in theory, but often not pos- sible in practice. The performance of zBW(kT) compared to the optimal noncausal ﬁlter in Table 2 is thus encouraging. 24T 20T 16T 12T 8T 4T 0T Time, t (s) 0 0.005 0.01 0.015 0.02 0.025 0.03 0.035 Figure 3: An example of the ﬁlter h(tm) given by (31), (16b), and T = 4. 5. APPLICATION EXAMPLE 1400 1200 1000 800 600 400 200 0 Time, t (s) 0 10 20 30 40 50 60 70 80 90 100 110 Instantaneous angular speed estimate, ω (rad/s) F 5 Th d t f h l d f Th d t i 10−1 10−2 Frequency, f (Hz) 10−4 10−3 10−2 10−1 100 H( f ) M = 160 M = 150 M = 140 M = 120 M = 100 Figure 4: The true ﬁlter, H( f ) (thick line), compared to \|E[ H( f )]\| for increasing values of M, when h(t) is given by (31). Figure 5: The data from a wheel speed sensor of a car. The data in the gray area is chosen for further evaluation. It includes more than 600 000 measurements. Figure 4: The true ﬁlter, H( f ) (thick line), compared to \|E[ H( f )]\| for increasing values of M, when h(t) is given by (31). Under these conditions, the work by [19] helps with opti- mally estimating z(kT). When estimating a function value z(kT) from a sequence u(tm) at times tm, a local weighted lin- ear approximation is investigated. The underlying function is approximated locally with a linear function Example 2. Data from a wheel speed sensor, like the one dis- cussed above, have been collected. An estimate of the angular speed, ω(t), Example 2. Data from a wheel speed sensor, like the one dis- cussed above, have been collected. An estimate of the angular speed, ω(t), m(t) = θ1 + (t −kT)θ2, (35) m(t) = θ1 + (t −kT)θ2, (35) (35) ω tm = 2π L tm −tm−1 , (32) (32) and m(kT) = θ1 is then found from minimization, and m(kT) = θ1 is then found from minimization, can be computed at every sample time. The average inter- sampling time, tM/M, is 2.3 milliseconds for the whole sam- pling set. The set is shown in Figure 5. We also indicate the time interval where the following calculations are performed. A sampling time T = 0.1 second gives a signal suitable for driver assistance systems. θ = argmin θ M m=1 u tm −m tm 2KB tm −kT , (36) (36) where KB(t) is a kernel with bandwidth B, that is, KB(t) = 0 for \|t\| > B. 5. APPLICATION EXAMPLE The Epanechnikov kernel, We begin with a discussion on ﬁnding the underlying sig- nal in an oﬄine setting and then continue with a test of dif- ferent online estimates of the wheel speed. KB(t) = 1 − t B 2 + , (37) (37) is the optimal choice for interior points, t1+B < kT < tM −B, both in minimizing MSE and error variance. Here, subscript + means taking the positive part. This corresponds to a non- causal ﬁlter for Algorithm 2. 5. APPLICATION EXAMPLE As a motivating example, consider the ubiquitous wheel speed signals in vehicles that are instrumental for all driver assistance systems and other driver information systems. The wheel speed sensor considered here gives L = 48 pulses per revolution, and each pulse interval can be converted to a wheel speed. With a wheel radius of 1/π, one gets 24ν pulses per second. For instance, driving at ν = 25 m/s (90 km/h) gives an average sampling rate of 600 Hz. This illustrates the need for speed-adaptive downsampling. where d f is the width of the ﬁlter. The sampling noise distri- bution is given by (16b). Figure 3 shows an example of the sampled ﬁlter h(tm). To produce a smooth causal ﬁlter, the time shift d f T is used. This in turn introduces a delay of d f T in the resampling procedure. We choose the scale factor d f = 8 for a better view of the convergence (higher d f gives slower convergence). The width of the ﬁlter covers approximately 2d f T/μT = 160 non- uniform samples, and more than 140 of them are needed for EURASIP Journal on Advances in Signal Processing 8 1400 1200 1000 800 600 400 200 0 Time, t (s) 0 10 20 30 40 50 60 70 80 90 100 110 Instantaneous angular speed estimate, ω (rad/s) Figure 5: The data from a wheel speed sensor of a car. The data in the gray area is chosen for further evaluation. It includes more than 600 000 measurements. 10−1 10−2 Frequency, f (Hz) 10−4 10−3 10−2 10−1 100 H( f ) M = 160 M = 150 M = 140 M = 120 M = 100 Figure 4: The true ﬁlter, H( f ) (thick line), compared to \|E[ H( f )]\| for increasing values of M, when h(t) is given by (31). 5.1. The optimal nonparametric estimate For the data set in Example 2, there is no true reference sig- nal, but in an oﬄine test like this, we can use computationally expensive methods to compute the best estimate. For this ap- plication, we can assume that the measurements are given by This gives the optimal estimate zopt(kT) = θ1, using the noncausal ﬁlter given by (35)–(37) with B = Bopt from [19], Bopt = 15σ2 C2MT/μT 1/5 . (38) u tm = s tm + e m (33) (33) (38) with 6. CONCLUSIONS This work investigated three diﬀerent algorithms for down- sampling non-uniformly sampled signals, each using inter- polation on diﬀerent levels. Two algorithms are based on ex- isting techniques for uniform sampling with interpolation in time and frequency domain, while the third alternative is truly non-uniform where interpolation is made in the con- volution integral. The results in the paper indicate that this third alternative is preferable in more than one way. (i) zE(kT): the casual ﬁlter given by (35), (36), the kernel (37) for −B < t ≤0 and B = 2Bopt; (i) zE(kT): the casual ﬁlter given by (35), (36), the kernel (37) for −B < t ≤0 and B = 2Bopt; (ii) zBW(kT): a causal Butterworth ﬁlter, h(t), in Algo- rithm 2; the Butterworth ﬁlter is of order 2 with cutoﬀ frequency 1/2T = 5 Hz, as deﬁned in (14), (iii) zm(kT): the mean of u(tm) for tm ∈[kT −T/2, kT + T/2]; (iv) zn(kT): a nearest neighbor estimate; Numerical experiments presented the root mean square error, RMSE, for the three algorithms, and convolution inter- polation has the lowest mean RMSE together with frequency- domain interpolation. It also has the lowest standard devia- tion of the RMSE together with time-domain interpolation. and compare them to the optimal zopt(kT). The last two estimates are included in order to show if the more clever estimates give signiﬁcant improvements. Figure 7 shows the ﬁrst two estimates, zE(kT) and zBW(kT), compared to the optimal zopt(kT). Theoretic analysis showed that the algorithm gives asymptotically unbiased estimates for noncausal ﬁlters. It was also possible to show how the actual ﬁlter implemented by the algorithm was given by a convolution in the frequency domain with the original ﬁlter and a window depending only on the sampling times. Table 2 shows the root mean square errors compared to the optimal estimate, zopt(kT), calculated over the interval indicated in Figure 5. From this, it is clear that the casual “optimal” ﬁlter, giving zE(kT), needs tuning of the band- width, B, since the literature gave no result for the opti- mal choice of B in this case. Both the ﬁltered estimates, zE(kT) and zBW(kT), are signiﬁcantly better than the sim- ple mean, zm(kT). with 654 653 652 651 650 649 648 647 Time, t (s) 78 78.2 78.4 78.6 78.8 79 79.2 79.4 79.6 79.8 Angular speed, ω (rad/s) 700 695 690 685 680 675 Time, t (s) 83 84 85 86 87 88 89 90 91 92 93 94 Angular speed, ω (rad/s) Figure 6: The cloud of data points, u(tm) black, from Example 2, and the optimal estimates, zopt(kT) gray. Only part of the shaded interval in Figure 5 is shown. Figure 7: A comparison of three diﬀerent estimates for the data in Example 2: optimal zopt(kT) (thick line), casual “optimal” zE(kT) (thin line), and causal Butterworth zBW(kT) (gray line). Only a part of the shaded interval in Figure 5 is shown. Table 2: RMSE from optimal estimate, E[\|zopt(kT) −z∗(kT)\|2], in Example 2. Table 2: RMSE from optimal estimate, E[\|zopt(kT) −z∗(kT)\|2], in Example 2. but KB(t) = (1 −t/B)+, t ∈[−B, 0] is optimal in MSE sense. Fan and Gijbels still recommend to always use the Epanech- nikov kernel, because of both performance and implemen- tation issues. [19] does not include a result for the optimal bandwidth in this case. In our test we need a causal ﬁlter and then choose B = 2Bopt in order to include the same number of points as in the noncausal estimate. the Epanechnikov kernel. It is promising that the estimate from Algorithm 2, zBW(kT), is close to zopt(kT), and it en- courages future investigations. 5.2. Online estimation The investigation in the previous section gives a reference value to compare the online estimates to. Now, we test four diﬀerent estimates: with (i) independent measurement noise, e(tm), with variance σ2, and In order to ﬁnd Bopt, the values of σ2, C, and μT were roughly estimated from data in each interval [kT −T/2, kT+T/2] and a mean value of the resulting bandwidth was used for Bopt. (ii) bounded second derivative of the underlying noise- free function s(t), that is, (ii) bounded second derivative of the underlying noise- free function s(t), that is, p The result from the optimal ﬁlter, zopt(kT), is shown compared to the original data in Figure 6, and it follows a smooth line nicely. s(2)(t) < C, (34) (34) which in the car application means limited accelera- tion changes. For end points, that is, a causal ﬁlter, the error variance is still minimized by said kernel, (37), restricted to t ∈[−B, 0], F. Eng and F. Gustafsson 9 9 700 695 690 685 680 675 Time, t (s) 83 84 85 86 87 88 89 90 91 92 93 94 Angular speed, ω (rad/s) Figure 6: The cloud of data points, u(tm) black, from Example 2, and the optimal estimates, zopt(kT) gray. Only part of the shaded interval in Figure 5 is shown. 654 653 652 651 650 649 648 647 Time, t (s) 78 78.2 78.4 78.6 78.8 79 79.2 79.4 79.6 79.8 Angular speed, ω (rad/s) Figure 7: A comparison of three diﬀerent estimates for the data in Example 2: optimal zopt(kT) (thick line), casual “optimal” zE(kT) (thin line), and causal Butterworth zBW(kT) (gray line). Only a part of the shaded inter al in Fig re 5 is sho n 700 695 690 685 680 675 Time, t (s) 83 84 85 86 87 88 89 90 91 92 93 94 Angular speed, ω (rad/s) Figure 6: The cloud of data points, u(tm) black, from Example 2, and the optimal estimates, zopt(kT) gray. Only part of the shaded interval in Figure 5 is shown. 654 653 652 651 650 649 648 647 Time, t (s) 78 78.2 78.4 78.6 78.8 79 79.2 79.4 79.6 79.8 Angular speed, ω (rad/s) Figure 7: A comparison of three diﬀerent estimates for the data in Example 2: optimal zopt(kT) (thick line), casual “optimal” zE(kT) (thin line), and causal Butterworth zBW(kT) (gray line). 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ACKNOWLEDGMENTS The authors wish to thank NIRA Dynamics AB for providing the wheel speed data, and Jacob Roll, for interesting discus- sions on optimal ﬁltering. Part of this work was presented at EUSIPCO07 The authors wish to thank NIRA Dynamics AB for providing the wheel speed data, and Jacob Roll, for interesting discus- i ti l ﬁlt i P t f thi k t d t The authors wish to thank NIRA Dynamics AB for providing the wheel speed data, and Jacob Roll, for interesting discus- sions on optimal ﬁltering. Part of this work was presented at EUSIPCO07 [17] S. R. Dey, A. I. Russell, and A. V. Oppenheim, “Precompen- sation for anticipated erasures in LTI interpolation systems,” IEEE Transactions on Signal Processing, vol. 54, no. 1, pp. 325– 335, 2006. sions on optimal ﬁltering. Part of this work was presented at EUSIPCO07 sions on optimal ﬁltering. Part of this work was presented at EUSIPCO07 [18] P. J. Ferreira, “Nonuniform sampling of nonbandlimited sig- nals,” IEEE Signal Processing Letters, vol. 2, no. 5, pp. 89–91, 1995. 6. CONCLUSIONS The Butterworth ﬁlter performs very well, and is also much less computationally complex than using p g In a ﬁnal example with empirical data, the algorithm gave signiﬁcant improvement compared to the simple local mean estimate and was close to the optimal nonparameteric esti- mate that was computed beforehand. EURASIP Journal on Advances in Signal Processing 10 Thus, the results are encouraging for further investiga- tions, such as approximation error analysis and search for optimality conditions. [16] M. Bourgeois, F. Wajer, D. van Ormondt, and F. Graveron- Demilly, “Reconstruction of MRI images from non-uniform sampling and its application to intrascan motion correction in functional MRI,” in Modern Sampling Theory, J. J. Benedetto and P. J. Ferreira, Eds., chapter 16, pp. 343–363, Birkh¨auser, Boston, Mass, USA, 2001. REFERENCES Ferreira, “Iterative and noniterative recovery of missing samples for 1-D band-limited signals,” in Sampling: Theory and Practice, F. Marvasti, Ed., chapter 5, pp. 235–282, Kluwer Academic Publishers, Boston, Mass, USA, 2001. [12] B. Lacaze, “Reconstruction of stationary processes sampled at random times,” in Nonuniform Sampling: Theory and Prac- tice, F. Marvasti, Ed., chapter 8, pp. 361–390, Kluwer Academic Publishers, Boston, Mass, USA, 2001. [13] H. G. Feichtinger and K. Gr¨ochenig, “Theory and practice of irregular sampling,” in Wavelets: Mathematics and Applica- tions, J. J. Benedetto and M. W. Frazier, Eds., pp. 305–363, CRC Press, Boca Raton, Fla, USA, 1994. [14] Y. C. Eldar, “Sampling with arbitrary sampling and recon- struction spaces and oblique dual frame vectors,” Journal of Fourier Analysis and Applications, vol. 9, no. 1, pp. 77–96, 2003. [15] K. Yao and J. B. Thomas, “On some stability and interpolatory properties of nonuniform sampling expansions,” IEEE Trans- actions on Circuits and Systems, vol. 14, no. 4, pp. 404–408, 1967.
https://openalex.org/W2023375468	https://europepmc.org/articles/pmc1464425?pdf=render	English	null	Genetic analysis of HLA, NA and HPA typing in type 2 diabetes and ASO	International journal of immunogenetics	2,006	cc-by	5,453	Summary We examined the genetic status of human leucocyte antigens (HLA), human platelet alloantigens (HPA) and neutrophil-speciﬁc antigens (NA) in patients with type 2 diabetes mellitus and diabetic arteriosclerosis obliterans (ASO). To our knowledge, the present study is the ﬁrst report showing the relationship among three genetic factors in type 2 diabetes mellitus and ASO patients. HLA typing was performed by the polymerase chain reaction (PCR)–restriction fragment length polymorphism method. HPA-typing and NA-typing were by a PCR-sequence-speciﬁc primer method. The incidence of HLA-DRB11501 was found to be signiﬁcant in type 2 diabetes and non-diabetic, particularly ASO-positive patients, compared to control subjects. There were no differences in NA1/NA2 between the control and diabetic or non-diabetic ASO groups. However, the frequency of NA2/NA2 in ASO-positive diabetes and non-diabetic ASO patients was signiﬁcantly higher than controls. The a/b genotype of HPA-5a/5b was signiﬁcantly lower in type 2 diabetes and non-diabetic ASO-positive patients than in controls. These ﬁndings suggest that genetic studies of HLA, NA and HPA could be useful to understand the pathogenesis of type 2 diabetes and ASO. Blackwell Publishing Ltd Genetic analysis of HLA, NA and HPA typing in type 2 diabetes and ASO S. Nomura, A. Shouzu,† S. Omoto,† T. Matsuzaki,‡ M. Yamaoka,‡ M. Abe,‡ M. Hosokawa,‡ M. Nishikawa,† T. Iwasaka† & S. Fukuhara* disease is steadily increasing among Asian populations (Kauffmann-Zeh et al., 2000). Epidemiological studies indicate that thrombotic diseases result from complex interactions between susceptibility factors, chronic en- vironmental inﬂuences and established intercurrent dis- orders such as diabetes mellitus (Kunicki, 2002). The critical role of platelets in the thrombotic process is now well accepted (Rauch et al., 2001). At sites of vascular injury, thrombus formation is initiated by binding platelet glyco- protein (GP) Ib to von Willebrand factor and binding platelet GPIa/IIa to collagen, which leads to platelet acti- vation and aggregation by binding platelet GPIIb/IIIa to ﬁbrinogen and von Willebrand factor (Kunicki, 2000). Several polymorphisms in the genes encoding for platelet GPs have been associated with increased platelet ad- hesiveness and aggregation, and in some studies, they have been associated with an increased risk of arterial throm- bosis (Bray, 1999; Bussel et al., 2000; Reiner et al., 2000). Human platelet alloantigens (HPA) are important for sensitization to platelet GP polymorphisms. In general, HPAs play important roles in the pathogenesis of neonatal alloimmune thrombocytopenia and post-transfusion pur- pura, as well as in some cases of refractoriness to platelet transfusion (Von dem Borne Kr. et al., 1981; Von dem Borne Kr. & Ouwehand, 1989; McFarland et al., 1991; Dreyfus et al., 1997). However, there are few reports available concerning the relationship between type 2 dia- betes and diabetic ASO. Correspondence: Shosaku Nomura, The First Department of Internal Medicine, Kansai Medical University, 10-15 Fumizonocho, Moriguchi, Osaka 570-8507, Japan. Tel: + 81 6 6992 1001; Fax: + 81 725 32 1113; E-mail: shosaku-n@mbp.ocn.ne.jp Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation. * The First Department of Internal Medicine, † The Second Department of Internal Medicine, ‡ Department of Blood Transfusion, Kansai Medical University, Osaka, Japan © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 Received 6 December 2004; revised 11 July 2005; accepted 22 December 2005 doi: 10.1111/j.1744-313X.2006.00581.x doi: 10.1111/j.1744-313X.2006.00581.x Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation. * The First Department of Internal Medicine, † The Second Department of Internal Medicine, ‡ Department of Blood Transfusion, Kansai Medical University, Osaka, Japan Received 6 December 2004; revised 11 July 2005; accepted 22 December 2005 Correspondence: Shosaku Nomura, The First Department of Internal Medicine, Kansai Medical University, 10-15 Fumizonocho, Moriguchi, Osaka 570-8507, Japan. Tel: + 81 6 6992 1001; Fax: + 81 725 32 1113; E-mail: shosaku-n@mbp.ocn.ne.jp Diagnosis of ASO All patients were evaluated by the ankle-pressure index (API). Patients with an API > 1.0 were considered not to have ASO. Thirty-three patients with API < 1.0 under- went angiography and plethysmography, and 28 were diagnosed as having ASO (Fontain’s class I–III). Of these, 21 patients with ASO underwent bypass procedures for femoral and trifurcation occlusive disease. Seventeen non- diabetic ASO patients were also diagnosed with Fontain’s class I–III. Of these, 10 patients with ASO underwent bypass procedures. Subjects We studied 104 Japanese patients (67 men and 37 women, aged 39–85 years) with type 2 diabetes mellitus, as deﬁned by the American Diabetes Association Criteria (The Expert Committee on the Diagnosis and Classiﬁca- tion of Diabetes Mellitus, 1997). Seventeen non-diabetic ASO patients were used for patient control subjects. A brief clinical proﬁle of the 104 diabetic patients and 17 non-diabetic ASO patients is presented in Table 1. One hundred and twenty-nine controls were randomly selected from among healthy unrelated Japanese individ- uals. Approval was obtained from the Institutional Review Board for these studies, and informed consent was obtained in accord with the Declaration of Helsinki. NA genotyping by PCR–preferential homoduplex formation assay (PHFA) We applied PCR–preferential homoduplex formation assay (PHFA) developed by Oka et al. (1994) to DNA typing of neutrophil-speciﬁc antigens (NA1, NA2 and SH) located on the neutrophil receptor, FcγRIIIb. Gene fragments containing the polymorphic sequences corre- sponding to NA1, NA2 and SH of FcγRIIIb, as well as the homologous fragment of FcγRIIIa, were ampliﬁed by PCR using speciﬁc primers (Fujiwara et al., 1996, 1999). Assay procedure was performed according to the manu- facturer’s instructions (Wakunaga Pharm. Co., Ltd, Hiro- shima, Japan). HLA DNA typing by PCR–RFLP Genomic DNAs from 104 diabetic patients, 17 non- diabetic patients and 126 healthy control subjects were Introduction With respect to the genetic aspects of thrombotic diseases such as type 2 diabetes mellitus, the HLA haplotype of patients can possibly be considered a potentially important factor in the aetiology, although its role remains unclear. This study examined the genetic status of HLA, HPA and NA in patients with type 2 diabetes mellitus and dia- betic ASO. To our knowledge, this is the ﬁrst report detail- ing relationships among these three genetic factors in type 2 diabetes mellitus and ASO patients. HPA DNA typing by enzyme-linked mini-sequence assay (ELMA) Gene fragments that contained polymorphic sequences corresponding to HPA-1, HPA-2, HPA-3, HPA-4, HPA-5 and HPA-6 were speciﬁcally ampliﬁed by the PCR method. Enzyme-linked mini-sequence assay (ELMA) typing was performed according to the manufacturer’s instructions (Smitest HPA Genotype ELMA, Sumitomo Metal Industries, Ltd, Ibaraki, Japan). Introduction Leucocytes have been suggested to play an important role in the development of atherosclerosis (Beekhuizen & van Furth, 1993; Jonasson et al., 1986). The ﬁrst step in the process of leucocyte inﬁltration into the sub-endothelial space is adhesion of circulating cells to the endothelium (Jonasson et al., 1986). Thus, the pathogenesis of athero- sclerosis appears to be inﬂammation (Ross, 1993). A recent study suggests genetic predisposition and chronic inﬂam- mation play the leading roles in the early stages and in the development of vascular damage (Diamantopoulos et al., 2002; Tsuchiya et al., 2003). In addition, the Fcγreceptor polymorphism is related to susceptibility of some immu- nological diseases (Dijstelbloem et al., 2000; Nieto et al., 2000; Fujimoto et al., 2001), and neutrophil-speciﬁc anti- gens (NA)1 and NA2 are located on the Fcγreceptor IIIb (Matsuo et al., 2000). On the other hand, studies have long suggested associations between certain human leuco- cyte antigens (HLA) and many autoimmune diseases. The underlying pathology of atherosclerosis obliterans (ASO) is basically atherosclerosis, and complete occlu- sions by fresh or old thrombi are often observed (Schwartz et al., 1995; Williams & Tabas, 1995). Recent evidence indicates that the incidence of thrombotic 117 © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 118 S. Nomura et al. isolated by phenol extraction of sodium dodecyl sulphate- lysed and proteinase K-treated cells. DNA was ampliﬁed by PCR with Taq DNA polymerase, and HLA typing was performed by the PCR–restriction fragment length poly- morphism (RFLP) method (Ota et al., 1992). The reaction mixture was subjected to 30 cycles of denaturation at 96–97 °C for 1 min, annealing at 55–62 °C for 1 min, and extension at 72 °C for 2 min in an automated PCR thermal sequencer (Iwaki Glass, Inc., Tokyo, Japan). After ampliﬁcation, aliquots of the products were digested with allele-speciﬁc restriction endonucleases for 3 h after addition of the appropriate reaction buffer. Samples of the ampliﬁed and cleaved DNAs were subjected to electrophoresis on 12% polyacrylamide gel in a minigel apparatus (Mupid-2, Cosmo Bio Co., Ltd, Tokyo, Japan). Cleavage of ampliﬁed fragments was detected by staining with ethidium bromide, and genotypes were determined on the basis of the RFLP band patterns thus generated. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 Statistical analysis Table 1. Clinical characteristics of the diabetic and non-diabetic patients Type 2 DM Non-diabetic ASO (–) ASO (+) ASO No. of patients 76 28 17 Sex ratio (male : female) 51 : 25 22 : 6 13 : 4 Age (year) Range 39–81 52–85 47–66 Median 54 66 59 BMI (kg m−2) Mean ± S.E. 28.1 ± 1.6 28.3 ± 2.1 24.2 ± 1.7 Table 1. Clinical characteristics of the diabetic and non-diabetic patients The χ2 method with continuity correction and Fisher’s exact test were used for HLA data analysis. Relative risk was calculated according to Wolf’s method with Hold- ane’s correction. Brieﬂy, it was calculated as (a × d)/(b × c), where a, b, c and d are the number of marker-positive patients, marker-negative patients, marker-positive con- trols and marker-negative controls, respectively (Thom- son, 1981). The frequencies of the FcγRIIIb and HPA alleles were estimated by a gene-counting method and tested for deviation from the Hardy–Weinberg equilib- rium using the χ2 test. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 HLA/NA/HPA in type 2 diabetes and ASO 119 119 Table 3. Results of genotyping of FcγRIIIB in healthy controls and patients Genotypes Healthy controls n = 129 Type 2 DM Non-diabetic ASO (–) n = 76 ASO (+) n = 28 ASO n = 17 NA1/NA1 48 (37.2%) 22 (28.9%) 3 (10.7%) 2 (11.7%) NA1/NA2 65 (50.4%) 36 (47.4%) 14 (50.0%) 8 (47.1%) NA2/NA2 16 (12.4%) 18 (23.7%) 11 (39.3%)a 7 (41.2%)b a P < 0.05 (χ2 = 4.570). b P < 0.05 (χ2 = 4.892). Table 3. Results of genotyping of FcγRIIIB in healthy controls and patients Table 2. Statistical analysis Frequencies of HLA-DRB1 alleles DRB1 Healthy controls (%) n = 129 Type 2 DM (%) Non-diabetic ASO (–) n = 76 ASO (+) n = 28 ASO n = 17 0101 17.8 13.2 10.7 11.8 0301 0.8 0 0 0 0401 0.8 0 0 0 0403 1.6 0 0 0 0404 0.8 0 0 0 0405 20.2 38.2 21.4 23.5 0406 13.2 5.3 3.6 5.9 0407 1.6 0 0 0 0410 4.7 2.6 3.6 5.9 0802 14.0 6.6 3.6 5.9 0803 17.8 19.7 17.9 23.5 0804 2.3 1.3 3.6 0 0901 20.2 22.4 17.9 17.6 1001 0.8 0 0 0 1101 2.3 5.3 3.6 5.9 1201 5.4 1.3 0 0 1202 0 2.6 0 0 1301 3.9 1.3 3.6 0 1302 11.6 6.6 7.1 11.8 1401 4.7 1.3 3.6 5.9 1402 0.8 0 0 0 1403 3.9 2.6 0 0 1405 4.7 1.3 0 0 1406 3.1 1.3 3.6 0 1407 0.8 0 0 0 1501 11.6 35.5a 60.7b 52.9c 1502 21.7 19.7 17.9 17.6 1601 1.6 1.3 3.6 5.9 1602 2.3 3.9 3.6 5.9 DM: diabetes mellitus. ASO: arteriosclerosis obliterans. a P < 0.05 (χ2 = 4.187). b P < 0.01 (χ2 = 11.745). c P < 0.05 (χ2 = 5.642). was found to be signiﬁcant in type 2 diabetes and non- diabetic ASO-positive patients compared to controls (con- trol vs. ASO-negative diabetic patients, 11.6% vs. 35.5%, P < 0.05; control vs. ASO-positive diabetic patients, 11.6% vs. 60.7%, P < 0.01; control vs. non-diabetic ASO-positive patients, 11.6% vs. 52.9%, P < 0.05). In addition, the incidence of this allele was found to be signiﬁcant in ASO patients compared to non-ASO patients, although there was no signiﬁcant difference between diabetic and non- diabetic patients (Fig. 1). On the other hand, another DRB1 allele did not exhibit signiﬁcant differences. Table 3 shows NA gene frequency in control and type 2 diabetes patients; there were no differences in NA1/ NA2 among the control, type 2 diabetes and non-diabetic ASO groups. However, the frequency of NA2/NA2 in ASO-positive diabetic and non-diabetic ASO patients was signiﬁcantly higher than in controls (control vs. ASO- negative diabetic patients, 12.4% vs. 23.7%, N.S.; control vs. ASO-positive diabetic patients, 12.4% vs. 39.3%, P < 0.05; control vs. non-diabetic ASO-positive patients, 12.4% vs. 41.2%, P < 0.05). Table 4 shows the HPA genotypes of the control and diabetic patients. Discussion Polymorphisms of class II genes within the major histo- compatibility complex can now be precisely determined by typing using the PCR–RFLP method. For example, serologically deﬁned HLA-DR antigens can be divided into many alleles reﬂecting the polymorphism of the amino acid sequence in the allelic hypervariable regions of the β chain. In addition, it has been reported that aplastic anemia patients with HLA-DR2 are likely to respond to immunosuppressive therapy (Nimer et al., 1994; Nakao et al., 1994). We have reported that the frequency of HLA-DR decreased and that of DRB10410 signiﬁcantly increased in immune thrombocytopenic purpura patients with poor response to prednisolone (Nomura et al., Figure 1. Relative frequencies of HLA-DRB11501 in type 2 diabetes and ASO patients. N.S. not signiﬁcant. Statistical analysis The a/b genotype of HPA-5a/5b was signiﬁcantly lower in type 2 diabetes and non-diabetic ASO patients than in controls (Control vs. ASO-negative diabetic patients, 7.0% vs. 1.3%, N.S.; control vs. ASO- positive diabetic patients, 7.0% vs. 0%, P < 0.05; control vs. non-diabetic ASO-positive patients, 7.0% vs. 0%, P < 0.05). Figure 1. Relative frequencies of HLA-DRB11501 in type 2 diabetes and ASO patients. N.S. not signiﬁcant. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 © 2006 The Authors Results For example, Raknes et al. (1998) reported that myasthenia gravis patients with the NA1/NA1 genotype had the most severe condition. The function of FcγRIIIb is not entirely clear; however, interestingly, FcγRIIIb induces actin ﬁla- ment assembly that is a prerequisite for phagocytosis, while cross-linking of the FcγRIIIb by immune complexes enhances FcγRII mediated phagocytosis (Salmon et al., 1991; Flesch et al., 1998). These functions may be related to the development of atherosclerosis, since the pathogen- esis of atherosclerosis includes inﬂammation. In addition, our results possibly reﬂect on such a hypothesis. 1998). Thus, the analysis of HLA antigens and alleles appears to offer some guidelines for the treatment of autoimmune diseases. In the present study, HLA DRB11501 was found to be more common in diabetic, particularly ASO-positive, patients. In addition, this allele was also found to be more common in non-diabetic ASO- positive patients. Thus, the association between patients and DRB11501 is not always related to type 2 diabetes, because this association is detected in non-diabetic patients. Type 1 diabetes was reported to be associated with some HLA alleles (Park, 2004). However, for the type 2 allele, such relationship was not detected. On the other hand, the association between HLA and infection is possible to detect. In the recent study, Veneri et al. (2005) reported the relationship between Helicobacter pylori and HLA in immune thrombocytopenic purpura patients. Our ﬁndings suggest the hypothesis that HLA regulates immu- noreactive inﬂammation or infection associated with type 2 diabetes or ASO. Thus, analysis of some HLA alleles may provide the information for predicting ASO or type 2 diabetes, if racial differences are taken into account. Diamantopoulos et al. (2002) evaluated 197 patients with type 2 diabetes, and found a signiﬁcant association between HLA-A3 and carotid intimal media thickness. In the Japanese population, the frequency of HLA-A3 is extremely low. In addition, most DRB11501 are associ- ated with HLA-A2, -A24 or -A26. The discrepancy between our results and Diamontopoulos’s is thought to depend on racial differences. However, most autoimmune diseases share pathogenetic mechanisms characterized by association with an HLA class II haplotype. Also, the pathogenesis of diabetic or non-diabetic ASO may be heterogeneous; therefore, analysis of a larger population with diverse genetic backgrounds will be needed. p y yp HPA plays an important role in provoking neonatal alloimmune thrombocytopenic purpura, post-transfusion purpura and refractoriness to platelet transfusion. Results Table 2 shows the HLA-DRB1 alleles in the control and type 2 diabetes patients. The incidence of HLA-DRB11501 © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 S. Nomura et al. 120 Table 4. Genotype frequencies of the six major HPAs Genotype Healthy controls n = 129 Type 2 DM Non-diabetic ASO (–) n = 76 ASO (+) n = 28 ASO n = 17 HPA-1a/1a (PlA1/PlA1) 100 (%) 100 (%) 100 (%) 100 (%) HPA-1a/1b (PlA1/PlA2) 0 0 0 0 HPA-1b/1b (PlA2/PlA2) 0 0 0 0 HPA-2a/2a (Kob/Kob) 73.6 84.2 82.1 82.4 HPA-2a/2b (Kob/Koa) 23.3 14.5 17.9 17.6 HPA-2b/2b (Koa/Koa) 3.1 1.3 0 0 HPA-3a/3a (Baka/Baka) 34.9 26.3 35.7 35.3 HPA-3a/3b (Baka/Bakb) 51.2 57.9 46.4 41.2 HPA-3b/3b (Bakb/Bakb) 14.0 15.8 17.9 23.5 HPA-4a/4a (Pena/Pena) 98.4 97.4 96.4 94.1 HPA-4a/4b (Pena/Penb) 1.6 2.6 3.6 5.9 HPA-4b/4b (Penb/Penb) 0 0 0 0 HPA-5a/5a (Brb/Brb) 93.0 98.7 100 100 HPA-5a/5b (Brb/Bra) 7.0 1.3 0 0* HPA-5b/5b (Bra/Bra) 0 0 0 0 HPA-6a/6a (Caa/Caa) 96.9 96.1 96.4 100 HPA-6a/6b (Caa/Cab) 3.1 3.9 3.6 0 HPA-6b/6b (Cab/Cab) 0 0 0 0 * P < 0.05 (χ2 = 5.625); P < 0.05 (χ2 = 4.492); * P < 0.05 (χ2 = 4.314). Table 4. Genotype frequencies of the six major HPAs FcγRIIIb is a glycosylphosphatidylinositol-anchored protein expressed on neutrophils (Ravetch & Perussia, 1989). NA1 and NA2 located on the neutrophil receptor FcγRIIIb are known to be involved in disorders such as neonatal alloimmune neutropenia, autoimmune neutro- penia and transfusion-related acute lung injury (Lalezari et al., 1960, 1975; Yomtovian et al., 1984). In addition, polymorphisms inﬂuencing the binding afﬁnity between the FcγRs and IgG of different subclasses are recently thought to be important for individual susceptibility to infection with gram-negative bacteria contributing to per- iodontal disease (Meisel et al., 2001). Indeed, Kobayashi et al. (2000) reported the importance of the FcγRIIIb- NA2 allele for susceptibility to IgG in Japanese periodon- titis patients. In the present study, the frequency of NA2/ NA2 was found to be signiﬁcantly higher in ASO-positive type 2 diabetes and non-diabetic ASO patients than in controls. Our results suggest the importance of the NA2 allele in susceptibility to ASO. Apart from some reports of periodontal disease, Fijen et al. (2000) reported that the FcγRIIIb-NA2/NA2 allotype appeared associated with meningococcal disease. However, previous reports have not always noted the importance of the NA2 allele. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 Results Diamantopoulos, E.J., Andreadis, E.A., Kakou, M.G., Diamantopoulos, E.J., Andreadis, E.A., Kakou, M.G., Vassilopoulos, C.V., Vlachonikolis, I.G., Gianna-Kopoulos, N.A., Tarassi, K.E., Papasteriades, C.A. & Nicolaides, A.N. (2002) Association of the HLA antigens with early atheromatosis in subjects with type 2 diabetes mellitus. International Angiology, 21, 379. Dijstelbloem, H.M., Bijl, M., Fijnheer, R., Scheepers, R.H., Oost, W.W., Jansen, M.D. et al. (2000) Fcγreceptor polymorphisms in systemic lupus erythematosus: association with disease and in vivo clearance of immune complexes. Arthritis and Rheumatism, 43, 2793. Dreyfus, M., Kaplan, C., Verdy, E., Schlegel, N., Durand-Zaleski, I. & Tchernia, G. (1997) Frequency of immune thrombocytopenia in newborns: a prospective study. Blood, 89, 4402. Fijen, C.A., Bredius, R.G., Kuijper, E.J., Out, T.A., De Haas, M., De Wit, A.P., Daha, M.R. & De Winkel, J.G. (2000) The role of Fcγreceptor polymorphisms and C3 in the immune defence against Neisseria meningitides in complement-deﬁcient individuals. Clinical and Experimental Immunology, 120, 338. Flesch, B.K., Maass, W. & Neppert, J. (1998) NA1/NA2 antisera inhibit FcγRI- but not FcγRII-mediated phagocytosis. Vox Sanguinis, 75, 247. Fujimoto, T.T., Inoue, M., Shimomura, T. & Fujimura, K. (2001) Involvement of Fc γreceptor polymorphism in the therapeutic response of idiopathic thrombocytopenic purpura. British Journal of Haematology, 115, 125. In conclusion, we performed a genetic analysis of HLA, NA and HPA in Japanese control subjects and type 2 dia- betes patients with and without ASO. The incidence of HLA-DRB11501 was found to be signiﬁcant in type 2 diabetes and non-diabetic ASO-positive patients com- pared with controls, and the frequency of this allele was signiﬁcant in patients with ASO. The frequency of NA2/ NA2 in ASO-positive diabetes and non-diabetic ASO- positive patients was signiﬁcantly higher than in controls. The a/b genotype of HPA-5a/5b was signiﬁcantly lower in type 2 diabetes and non-diabetic ASO-positive patients than in controls. These ﬁndings suggest that genetic studies of HLA, NA and HPA could be useful to understand the pathogenesis of type 2 diabetes and ASO. Fujiwara, K., Isa, K., Oka, T., Maekawajiri, S., Yamane, A., Akaza, T., Tadokoro, K., Juji, T., Shibata, Y. & Tokunaga, K. (1996) Large-scale DNA typing for human platelet alloantigens by PCR–RHFA (preferential homoduplex-formation assay). British Journal of Haematology, 95, 198. Fujiwara, K., Watanabe, Y., Mitsushige, S., Oka, T., Yamane, A., Akaza, T., Tadokoro, K., Tokunaga, K., Shibata, Y. & Juji, T. Acknowledgements This study was supported in part by a Research Grant for Advanced Medical Care from the Ministry of Health and Welfare of Japan. Jonasson, L., Holm, J., Bondjers, G. & Hansson, G.K. (1986) Regional accumulation of T cells, macrophages, and smooth muscle cells in the human atherosclerotic plaque. Arteriosclerosis, 6, 131. Results (1999) Determination of granulocyte-speciﬁc antigens on neutrophil Fcγreceptor IIIb by PCR–preferential homoduplex formation assay, and gene frequencies in the Japanese population. Vox Sanguinis, 77, 218. Gonzalez-Conejero, R., Lozano, M.L., Rivera, J., Corral, J., Iniesta, J.A., Moraleda, J.M. & Vicente, V. (1998) Polymorphisms of platelet membrane glycoprotein Ibα associated with arterial thrombotic disease. Blood, 92, 2771. Jilma-Stohlawetz, P., Homoncik, M., Jilma, B., Knechtelsdorfer, M., Unger, F., Mannhalter, C., Santoso, S. & Panzer, S. (2003) Glycoprotein Ib polymorphisms inﬂuence platelet plug formation under high shear rates. British Journal of Haematology, 120, 652. Results To date, 10 HPA systems have been described and several additional alloantigen systems have been reported (Bussel et al., 2000; Kunicki, 2000, 2002). In particular, HPA-1 (PlA.), HPA-2 (Ko), HPA-3 (Bak), HPA-4 (Pen) and HPA- 5 (Br) have been well established by serological analysis (Kunicki & Newman, 1992). The HPA-1a (PlA1, Zwa) antigen is by far the most common antigen implicated in neonatal alloimmune thrombocytopenia in a Caucasian population (Taaning et al., 1983; McFarland et al., 1989; Mueller-Eckhardt et al., 1989). On the other hand, HPA- 4b is responsible for neonatal alloimmune thrombocyto- penia in Japanese. This discrepancy in ﬁndings would have occurred because the HPA phenotype frequency varies between races. HPA systems also reside on functional platelet receptor proteins. It has been shown by a number of groups that all platelet-speciﬁc alloantigens thus far dis- covered are located on four platelet membrane GPs: GPIIb, GPIIIa, GPIb and GPIa. Thus, it has been reported that polymorphism of HPA is a genetic risk factor for thrombotic diseases (Weiss et al., 1996; Gonzalez- Conejero et al., 1998; Sperr et al., 1998; Carter et al., 1999; Kroll et al., 2000; Baker et al., 2001; Mikkelsson et al., © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 HLA/NA/HPA in type 2 diabetes and ASO 121 2001, 2002; Jilma-Stohlawetz et al., 2003). In the present study, the a/b genotype of HPA-5a/5b was signiﬁcantly lower in type 2 diabetes and non-diabetic ASO patients than in controls. The Lys505Glu amino acid substitution located in the cation-binding domain of α2 is responsible for HPA-5b (Bra) and HPA-5a (Brb) epitope formation (Santoso et al., 1993). In a large study, Kroll et al. (2000) found an association between HPA-5 dimorphism in a low-risk patient subgroup with coronary artery disease. In this population, the frequency of Brb homozygous individ- uals was overrepresented. This ﬁnding suggests that the allele 3 (807C; Bra) may increase the risk for thrombotic disease through a quantitative effect on the function that is independent of the genetic effects on expression levels. However, the frequency of phenotype HPA-5b in Japa- nese is lower than that in Caucasian (8.7 vs. 20.6%). In addition, it has been reported that the probability of HPA- related immunization is strongly associated with HLA class II DR types (Gonzalez-Conejero et al., 1998). A larger number of patients will need to be examined to conﬁrm this possibility. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 References & Stroncek, D. (2000) Variations in genes encoding neutrophil antigens NA1 and NA2. Transfusion, 40, 645. Ross, R. (1993) The pathogenesis of atherosclerosis: a prospective study for the 1990s. Nature, 362, 801. McFarland, J.G., Aster, R.H., Bussel, J.B., Gianopoulos, J.G., Derbes, R.S. & Newman, P.J. (1991) Prenatal diagnosis of neonatal alloimmune thrombocytopenia using allele-speciﬁc oligonucleotide probes. Blood, 78, 2276. Salmon, J.E., Brogle, N.L., Edberg, J.C. & Kimberly, R.P. (1991) Fcγreceptor III induces actin polymerization in human neutrophils and primers phagocytosis mediated by Fcγ receptor II. Journal of Immunology, 146, 997. McFarland, J.G., Frenzke, M. & Aster, R.H. (1989) Testing of maternal sera in pregnancies at risk for neonatal alloimmune thrombocytopenia. Transfusion, 29, 128. Santoso, S., Kalb, R., Walka, V., Kiefel, V., Mueller-Eckhardt, C. & Newman, P.J. (1993) The human platelet alloantigens Br(a) and Br(b) are associated with a single amino acid polymorphism on glycoprotein Ia (integrin subunit α2). Journal of Clinical Investigations, 92, 2427. Meisel, P., Carlsson, L.E., Sawaf, H., Fangjaenal, J., Greinacher, A. & Kocher, T. (2001) Polymorphisms of Fcγ-receptors RIIa, RIIIa, and RIIIb in patients with adult periodontal diseases. Genes Immunology, 2, 258. Schwartz, S.M., De Blois, D. & O’Brien, E.R. (1995) The intima. Soil for atherosclerosis and restenosis. Circulation Research, 77, 445. Mikkelsson, J., Perola, M., Penttila, A. & Karhunen, P.J. (2001) Platelet glycoprotein IbαHPA-2 Met/VNTR B haplotype as a genetic predictor of myocardial infarction and sudden cardiac death. Circulation, 104, 876. Sperr, W.R., Huber, K., Roden, M., Janisiw, M., Lang, T., Graf, S., Maurer, G., Mayr, W.R. & Panzer, S. (1998) Inherited platelet glycoprotein polymorphisms and a risk for coronary heart disease in young central europeans. Thrombosis Research, 90, 117. Mikkelsson, J., Perola, J., Penttila, A. & Karhunen, P.J. (2002) Platelet collagen receptor GPIa (807T/HPA-5) haplotype is associated with an increased risk fatal coronary events in middle-aged men. Atherosclerosis, 165, 111. Taaning, E., Antonsen, H., Petersen, S., Svejgaard, A. & Thomsen, M. (1983) HLA antigens and maternal antibodies in alloimmune neonatal thrombocytopenia. Tissue Antigens, 21, 351. Mueller-Eckhardt, C., Kiefel, V., Grubert, A., Kroll, H., Weisheit, M., Weisheit, M., Schmidt, S., Mueller-Eckhardt, G. & Santoso, S. (1989) 348 cases of suspected neonatal alloimmune thrombocytopenia. Lancet, I, 363. The Expert Committee on the Diagnosis and Classiﬁcation of Diabetes Mellitus (1997) Report of the expert committee on the diagnosis and classiﬁcation of diabetes mellitus. Diabetes Care, 20, 1183. References Baker, R.I., Eikelboom, J., Lofthouse, E., Staples, N., Afshar-Khaghan, V., Lopez, J.A., Shen, Y., Berndt, M.C. & Hankey, G. (2001) Platelet glycoprotein Ibα Kozak polymorphism is associated with an increased risk of ischemic stroke. Blood, 98, 36. Kauffmann-Zeh, A., Dhand, R. & Allen, L. (2000) Vascular biology. Nature, 407, 219. Kobayashi, T., Sugit, N., van der Pol, W.L., Nunokawa, Y., d l A d k l G Kobayashi, T., Sugit, N., van der Pol, W.L., Nunokawa, Y., Westerdaal, N.A., Yamamoto, K., van de Winkel, J.G. & Yoshie, H. (2000) The Fcγreceptor genotype as a risk factor for generalized early-onset periodonitis in Japanese patients. Journal of Periodontology, 71, 1425. Beekhuizen, H. & van Furth, R. (1993) Monocyte adherence to human vascular endothelium. Journal of Leukocyte Biology, 54, 363. Kroll, H., Gardemann, A., Fechter, A., Haberbosch, W. & Santoso, S. (2000) The impact of the glycoprotein Ia collagen receptor subunit A1648G gene polymorphism on coronary artery disease and acute myocardial infarction. Thrombosis and Haemostasis, 83, 392. Bray, P.F. (1999) Integrin polymorphisms as risk factors for thrombosis. Thrombosis and Haemostasis, 82, 337. Bussel, J.B., Kunicki, T.J. & Michelson, A.D. (2000) Platelets: new understanding of platelet glycoproteins and their role in disease. Hematology (American Society of Hematology Education Program) 222. Kunicki, T.J. (2002) The inﬂuence of platelet collagen receptor polymorphisms in hemostasis and thrombotic disease. Arteriosclerosis, Thrombosis, and Vascular Biology, 22, 14. Carter, A.M., Catto, A.J., Bamford, J.M. & Grant, P.L. (1999) Association of the platelet glycoprotein Iib HPA-3 polymorphism with survival after acute ischemic stroke. Stoke, 30, 2606. Kunicki, T.J. & Newman, P.J. (1992) The molecular immunology of platelet proteins. Blood, 80, 1386. © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 S. Nomura et al. 122 FcRIII (CD16) on human natural killer cells and neutrophils: cell type-speciﬁc expression of two genes that differ in single nucleotide substitutions. Journal of Experimental Medicine, 170, 481. Lalezari, P., Jiang, A.F., Yegen, L. & Santorineou, M. (1975) Chronic autoimmune neutropenia due to anti-NA2 antibody. New England Journal of Medicine, 293, 744. Lalezari, P., Nussbaum, M., Gelman, S. & Spaet, T.H. (1960) Neonatal neutropenia due to maternal isoimmunization. Blood, 15, 236. Reiner, A.P., Kumar, P.N., Schwartz, S.M., Longstreh, W.T., Pearce, R.M., Rosendaal, F., Psaty, B.M. & Siscovick, D.S. (2000) Genetic variants of platelet glycoprotein receptors and risk of stroke in young women. Stroke, 31, 1628. Matsuo, K., Procter, J. References Nakao, S., Takamatsu, H., Chuhjo, T., Ueda, M., Shiobara, S., Matsuda, T., Kaneshige, T. & Mizoguchi, H. (1994) Identiﬁcation of a speciﬁc HLA class II haplotype strongly associated with susceptibility to cyclosporine-dependent aplastic anemia. Blood, 84, 4257. Thomson, G. (1981) A review of theoretical aspect of HLA and disease association. Theoretical Population Biology, 20, 168. Tsuchiya, N., Kobayshi, S., Kawasaki, A., Kyogoku, C., Arimura, Y., Yoshida, M., Tokunaga, K. & Hashimoto, H. (2003) Genetic background of Japanese patients with antineutrophil cytoplasmic antibody- associated vasculitis: association of HLA-DRB10901 with microscopic polyangitis. Journal of Rheumatology, 30, 1534. Nieto, A., Caliz, R., Pascual, M., Mataran, L., Garcia, S. & Martin, J. (2000) Involvement of Fcγreceptor IIIA genotypes in susceptibility to rheumatoid arthritis. Arthritis and Rheumatism, 43, 735. Nimer, S.D., Ireland, P., Meshkinpour, A. & Frane, M. (1994) An increased HLA DR2 frequency is seen in aplastic anemia patients. Blood, 84, 923. Veneri, D., De Matters, G., Solero, P., Federici, F., Zanuso, C., Guizzardi, E., Arena, S., Gaio, M., Pontiero, P., Ricetti, M.M. & Franchini, M. (2005) Analysis of B- and T-cell clonality and HLA class II alleles in patients with idiopathic thrombocytopenic purpura: correlation with Helicobacter pylori infection and response to eradication treatment. Platelets, 16, 367. Nomura, S., Matsuzaki, T., Ozaki, Y., Yamaoka, M., Yoshimura, C., Katsura, K., Xie, G.L., Kagawa, H., Ishida, T. & Fukuhara, S. (1998) Clinical signiﬁcance of HLA-DRB1*0410 in Japanese patients with idiopathic thrombo-cytopenic purpura. Blood, 91, 3616. Von dem Borne, A.E.G. Kr, van Leeuwen, E.F., von Riesz, L.E., van Boxtel, C.J. & Engelfriet, C.P. (1981) Neonatal alloimmune thrombocytopenia: detection and characterization of the respomsible antibodies by the platelet immunoﬂuorescence test. Blood, 57, 649. Oka, T., Matsunaga, H., Tokunaga, K., Mitsunaga, S., Juji, T. & Yamane, A. (1994) Simple method for detecting single base substitutions and its application to HLA-DPB1 typing. Nucleic Acids Research, 22, 1541. Von dem Borne, A.E.G. Kr & Ouwehand, W.H. (1989) Immunology of platelet disorders. Bailliere’s Clinical Haematology, 2, 749. Ota, M., Seki, T., Fukushima, H., Tsuji, K. & Inoko, H. (1992) HLA DRB1 genotyping by modiﬁed PCR-RFLP method combined with group-speciﬁc primers. Tissue Antigens, 39, 187. Weiss, E.J., Bray, P.F., Tayback, M., Schulman, S.P., Kicker, T.S., Becker, L.C., Weiss, J.L., Gerstenblith, G. & Goldschmidt-Clermont, P.J. (1996) A polymorohism of a platelet glycoprotein receptor as an inherited risk factor for coronary thrombosis. New England Journal of Medicine, 334, 1090. Park, Y. References (2004) Prediction of the risk of type 1 diabetes from polymorphisms in candidate genes. Diabetes Res Clin Pract, 66, S19. Raknes, G., Skeie, G.O., Gilhus, N.E., Aadland, S. & Vedeler, C. (1998) FcγRIIA and FcγRIIIB polymorpjisms in myasthenia gravis. Journal of Neuroimmunology, 81, 173. Williams, K.J. & Tabas, I. (1995) The response-to-retention hypothesis of early atherosclerosis. Arteriosclerosis, Thrombosis, and Vascular Biology, 15, 551. Rauch, U., Osende, J.I., Fuster, V., Badimon, J.J., Fayad, Z. & Chesebro, J.H. (2001) Thrombus formation on atherosclerotic plaques: pathogenesis and clinical consequences. Annals of Internal Medicine, 134, 224. Yomtovian, R., Press, C., Engman, H., Kline, W., Clay, M., Hammer-Schmidt, D. & McCullough, J. (1984) Severe pulmonary hypersensitivity associated with passive transfusion of a neutrophil-speciﬁc antibody. Lancet, 1, 244. Ravetch, J.V. & Perussia, B. (1989) Alternative membrane forms of © 2006 The Authors Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122 Journal compilation © 2006 Blackwell Publishing Ltd, International Journal of Immunogenetics 33, 117–122
https://openalex.org/W2075569624	https://zenodo.org/records/1982056/files/article.pdf	English	null	ABSTRACT OF AN Address ON MEDICAL ETHICS.	Lancet	1,904	public-domain	3,172	This etiquette is indeed only a branch of the salutary discipline which society in general exercises over its individual members, and the restraint which our etiquette places upon those who follow their own supposed interests, regardless of the injury which they may inflict on their brethren, is a wholesome restraint and has been enforced by this society with a due measure of leniency and justice A, lymph record from purin-free meal. B, purln-free meal with sodium chloride 2’0 grammes. c, purin-free meal with sodium chloride 4’0 grammes. D, purin-free meal with sodium chloride 2 grammes and uric acid 0’016, creatin 0’033. creatinin 0’033, xanthin 0’016, hypoxanthin 0’016, and glycogen 0048. A, lymph record from purin-free meal. B, purln-free meal with sodium chloride 2’0 grammes. c, purin-free meal with sodium chloride 4’0 grammes. D, purin-free meal with sodium chloride 2 grammes and uric acid 0’016, creatin 0’033. creatinin 0’033, xanthin 0’016, hypoxanthin 0’016, and glycogen 0048. It seems to me that these facts demonstrate that such bodies as uric acid, creatin, creatinin, xanthin, glycogen, and sodium chloride perform an important function in nutrition, for during digestion they act as distributers of lymph to all the tissues, an office which the nutrient con- stituents themselves (proteids, fats, and carbohydrates) are incapable of discharging. As inquiry goes forward it may discover other substances possessing this physiological property associated with our food elements. Nature has, with a wise intent, widely distributed these active chemical agents through most of our food-stuffs and beverages, thus turning even her waste products to the best account, the nutrition of the organism. Unfortunately, man, in his efforts to improve on nature, so often turns these benefactors into malefactors. Besides these lymph factors introduced from without there are supplies of the same materials generated within us. It will be shown, too, that exercise, sleep, and variations of temperature are important physio- logical conditions for the autogenous supply of tissue lymph. It is not improbable that in some subjects the endogenous production of lymph factors may exceed the physiological average, when an adjustment can only be effected by a limitation of the exogenous supply. But I am here touching the borderland between physiology and pathology, to which I hope to revert in the second lecture. justice. 1180 1180 speculations on the philosophical basis of ethics and pro. ceeded as follows :- addition of salt and more especially by that of salt asso- ciated with the physiological products than when taken alone. The great function of a medico-ethical association appears to me to be to assist in applying the rule of right in medical matters. It will, therefore, naturally endeavour (1) to impose a wholesome restraint upon the liberty of individual members of the profession in the interest of all, and (2) to assist in passing wise laws in those matters which relate specially to public health and the interests of the profession. FIG. 5. FIG. 5. A, lymph record from purin-free meal. B, purln-free meal with sodium chloride 2’0 grammes. c, purin-free meal with sodium chloride 4’0 grammes. D, purin-free meal with sodium chloride 2 grammes and uric acid 0’016, creatin 0’033. creatinin 0’033, xanthin 0’016, hypoxanthin 0’016, and glycogen 0048. ] profession. Let me first say a few words upon the restraints which the Medico-Ethical Association of Manchester has imposed upon the conduct of its own members. The chief restraint which finds expression in our laws relates to the prosaic and homely, but by no means unimportant, question of fees. It is as essential for the public as for us that our dignity should be maintained and we give to them the greatest guarantee that they will have honest and trustworthy services in the time of need by showing that we are a self-respecting and a disinterested profession. This society has been performing a public duty by insisting that its members shall claim fair remuneration for services rendered. That our tariff which has been drawn up by the society is a fair one is attested by the fact that it forms the basis of every scale of fees now in existence in this country. Our association has also done much to raise the character of the profession by inculcating a high code of honour in the dealings of members with one another. The laws of this code embody implicitly the hitherto unwritten rules which constitute the etiquette of the profession. ABSTRACT OF AN Address ON MEDICAL ETHICS. Being the Presidential (Valedictory) Address delivered before the Medico-Ethical Association of Manchester on Jan. 27th, 1904, BY D. LLOYD ROBERTS, M.D. ST. AND., F.R C.P. LOND., PHYSICIAN TO ST. MARY’S HOSPITAL, MANCHESTER; CONSULTING OBSTETRIC PHYSICIAN TO THE MANCHESTER ROYAL INFIRMARY; AND LECTURER ON CLINICAL OBSTETRICS AND GYNÆCOLOGY AT THE OWENS COLLEGE, MANCHESTER. ABSTRACT OF AN Address ON MEDICAL ETHICS. Being the Presidential (Valedictory) Address delivered before the Medico-Ethical Association of Manchester on Jan. 27th, 1904, BY D. LLOYD ROBERTS, M.D. ST. AND., F.R C.P. LOND., PHYSICIAN TO ST. MARY’S HOSPITAL, MANCHESTER; CONSULTING OBSTETRIC PHYSICIAN TO THE MANCHESTER ROYAL INFIRMARY; AND LECTURER ON CLINICAL OBSTETRICS AND GYNÆCOLOGY AT THE OWENS COLLEGE, MANCHESTER. Many schemes of social advancement and reform which at the time of their inception were advocated in this very room have already found, and others will yet find, their embodiment in the laws of our country. The question of provident dispensaries was advocated in a paper read before the society nearly 50 years ago. Other subjects discussed in the early years of this society were medical education and the necessity of turning hospitals to good account for pur- poses of teaching. In 1869 this society advocated a reform of the lunacy laws. I need not tell you that from the point of view of the medical profession the lunacy laws are not satisfactory and I believe that the public are no more contented than we are with the present condition of these laws. In 1870 the "one-portal" system was discussed and pressed upon the Lord President of the Council. Shortly afterwards the direct representation of the profession in the Medical Council and the appointment of a Public Prosecutor were the subjects of interesting debate. Amongst other questions of interest which underwent discussion may be mentioned the means of notifying to sanitary authorities the occurrence of infectious disease, the duty of corporate bodies to provide for the isolation of infectious disease, the working of the Factory Acts, and the position of ship surgeons. tEYESTRAIN AS A CAUSE OF HEADACHE AND OTHER NEUROSES. Again and again is the same story told, first of the suffering and hen of the relief afforded by optical aid. In my last 800 efraction cases in private practice I find that 162 suffered rom headaches. I do not include in this number those oaking the usual complaint of discomfort for which hyper- netropes and early presbyopes so frequently solicit assist- ance. In the cases to which I refer the patients have at ance volunteered that they suffered from severe headaches or lave stated on inquiry that they did do so. In the great najority this constituted the cause for seeking advice. In nost, also, in varying degrees, were associated some or all .f the train of symptoms to be described later. practise Opinions differ as to how far a government is justified in enforcing the practice of vaccination, in insisting upon absolute compulsory isolation in cases of infectious diseases, and in compelling corporations, property owners, and th public generally to observe wise sanitary laws. I do not believe that this country will ever turn its back upon the Vaccination Acts, but it will be well for the profession to be in readiness to advise the Government and the constituencies in this matter. In reference to the general isolation of the acute infective diseases, I believe it will be possible to obtain even more stringent regulations than we now possess as the people become more educated without the Government having to trench too violently upon the liberty of the subject. Sanitary regulations will be more carefully observed under the pressure of an educated public opinion without our being too ready to call out for coercive legislative enactments. Stringent laws, like every species of coercion, have a tendency to defeat themselves. A drastic sanitary law would at once call into being an anti-sanitation societ and it is only on condition that there was a strong educated and even uneducated public opinion in favour of the law that it would maintain its place upon the statute book. p I b a , g, n n h e ch o d t a n ae n n v ee t, a r ; ne nm i, r a r r e symptoms Perhaps no more important work has been accomplished n the whole range of medicine and surgery than that of )lacing anomalies of refraction on a firm scientific basis. tEYESTRAIN AS A CAUSE OF HEADACHE AND OTHER NEUROSES. BY SIMEON SNELL, F.R.C.S. EDIN., OPHTHALMIC SURGEON, SHEFFIELD ROYAL INFIRMARY; PROFESSOR OF OPHTHALMOLOGY, UNIVERSITY COLLEGE, SHEFFIELD. m s- ,tl n. sl r . on n o: OF all painful affections probably headache causes the lost distress and misery. The individual who never ex- eriences headache is one much to be envied. Fortunately bere are many able to say this. On the other hand, there re many afflicted with headache causing constant or re- urring suffering and only too frequently so disabling the offerer that life to him or her becomes a burden. Life is horn of its beauties and not only are its pleasures curtailed ut the actual daily occupation is interfered with. A con- ideration of some of the causes of this suffering and the neasures which may be adopted for its relief is well worthy f our attention. great circumspection. Much has been said regarding admission into the profe sion by the "one-portal" system and various views have been held by highly respectable authorities on this questio We have at present about 23 licensing bodies. The practical question is, Shall we abolish the existing powers of these institutions ?-or should we content ourselves with enacting guarantees that they shall maintain the standard of thei examinations at a sufficiently high level of attainments ? In any scheme of State examination the standard of examinati must be a minimum standard and the lowest prizes in the profession must fix it. The Government requires medical men for the navy and for the army and it would be impossible to make the standard of examinations so hig as to exclude men who would be likely to accept such appointments. There are also parts of the country with widely scattered populations and no government will raise the standard of examinations so high that men will not be found to practise in those districts. h g a f] t] n a t] r,e 13 n a o h n n o It has, of course, for some time been understood that ertain instances of headache bear some relation to ye defects. But the far-reaching effects of eyestrain ,nd the relief afforded by the proper correction of re- raction errors are not by any means as widely recognised as hey demand. It is a matter that, in my opinion, admits of no question. The evidence is continually before one. tEYESTRAIN AS A CAUSE OF HEADACHE AND OTHER NEUROSES. Chis we owe in the greatest measure to Donders and it )rought to him undying fame. It is well known, too, that astigmatism, which has such a close bearing on our subject, vas discovered by Thomas Young, the physician as well as he physicist. Sir George Airy, the astronomer royal, de- cribed astigmatism in his own case. It was of the compound nyopic variety and was monocular. Here it is of interest to note that Airy was the subject of migraine and he described his own case in a paper in the Philosophical Magazine or 1865.2 It may be remarked in passing that a proper :orrection of his refraction error, with the knowledge which ve now possess, would in all probability have relieved him f his migraine, for he had both astigmatism and asymmetry n the refraction of the eyes, two very potent causes of head- ache as will be shown farther on. Airy’s son, Dr. Hubert Ury, also published a very graphic description of migraine md the ocular symptoms in his own person and it appears lot unlikely from the account which he gives that be, too, vould have been relieved, as has so often been the case in my ixperience in similar instances, if attention had been given o his eyes.4 The interest that Young and Airy had in astigmatism was, however, physiological and not patho- ogical. And even Donders lays stress always on the lormal and abnormal astigmatism, the latter being astig- natism which interfered with the acuteness of vision. Our nterest here is in the pathological significance of astig- natism and the subtle and powerful influences of eyestrain md not necessarily on its interference with visual acuteness. Frequently no complaint of vision is made and the error of ’efraction is only discovered after a careful and complete examination of the eyes. place upon Another subject which has received much public attention of late is over-pressure in schools. It is manifest that the pupils must possess a certain amount of intelligence befor any teaching can be effective, and their faculties cannot be maintained and developed unless they are supplied wit plenty of nourishing food and kept under healthy sanitary conditions. Underfed children cannot have well-develop faculties ; therefore, they are not in the position to benefi fully by teaching, however perfect the method of instructi may be. n - ‘ i - , 1 A paper read before the York Medical Society on Feb. 24th, 1904. 2 Philosophical Magazine, July, 1865, vol. xxx., p. 19. 3 Transactions of the Royal Society, 1870. 4 He speaks of his attacks coming on after close eye-work, and says, ’ Usually after two or three hours’ close reading, especially if I had insufficient exercise, I become aware that part of the letter I am looking at or a word at some little distance from the sight point is eclipsed by a dim cloud spot, &c." "Want of exercise, sedentary employment, close reading and writing are the usual antecedents. It generally comes on while the eyes are engaged with toilsome reading." 1181 1181 with their numerous members practising all over the country, should not be represented in Parliament. We are no politically powerless. Although we do not care to take an active part in political agitations yet we greatly influence public opinion, and I am not sure whether our political power might not be diminished rather than increased by direct representation of the profession in Parliament. Action begets reaction and it is probable that a corporate repre- sentation would tell against us rather than in our favour. I have already advocated the appointment of a Minister of Public Health, but the powers intrusted to him ought to be exercised with great circumspection. with their numerous members practising all over the country, should not be represented in Parliament. We are no politically powerless. Although we do not care to take an active part in political agitations yet we greatly influence public opinion, and I am not sure whether our political power might not be diminished rather than increased by direct representation of the profession in Parliament. Action begets reaction and it is probable that a corporate repre- sentation would tell against us rather than in our favour. I have already advocated the appointment of a Minister of Public Health, but the powers intrusted to him ought to be exercised with great circumspection tEYESTRAIN AS A CAUSE OF HEADACHE AND OTHER NEUROSES. BY SIMEON SNELL, F.R.C.S. EDIN., OPHTHALMIC SURGEON, SHEFFIELD ROYAL INFIRMARY; PROFESSOR OF OPHTHALMOLOGY, UNIVERSITY COLLEGE, SHEFFIELD. n 1 A paper read before the York Medical Society on Feb. 24th, 1904. y hilosophical Magazine, July, 1865, vol. xxx., p. 19. 3 Transactions of the Royal Society, 1870. ABSTRACT OF AN Address ON MEDICAL ETHICS. surgeons. I would now direct your attention to the desirability of having a Minister of Public Health. A governmental sanitary department will always have plenty of work in dealing with the problems arising from overcrowding in cities, such as the limitation of infectious diseases, the disposal of sewage, and many other questions which directly relate to the public health. Many years ago in this society attention was directed to the benefit that the State would derive were the medical profession represented in the House of Peers. This is now an accomplished fact. All honour to the distinguished statesman who was privileged to advise the Crown to elevate Lord Lister, the first medical peer, to a seat in the House of Lords. Surely no one is more capable to give advice to a government on a variety of subjects than a highly educated and experienced medical man. Medicine is based upon many sciences, and our profession is the only one which possesses a wide knowledge of the conditions on which much of the comfort and happiness of life depends IN the course of his introductory remarks Dr. Lloyd Roberts said that the Medico-Ethical Association of Man- chester was the first organisation of the kind which was established and that the celebrated "Medical Ethics" of Thomas Percival was written in Manchester rather more than a hundred years ago. He referred briefly to various happiness depends. Much has been said of late years of the political power- lessness of the medical profession and while not altogether sympathising with this cry I see no reason why influential corporations like the Colleges of Physicians and Surgeons, 1181 tEYESTRAIN AS A CAUSE OF HEADACHE AND OTHER NEUROSES. In order to meet this difficulty it has been pro- posed to supply the children in poor districts with a fre midday meal. This scheme must meet with our approbatio so far as the poor children themselves are concerned, but we ought to insist, in my opinion, that the project should b carried out by the payment of the parents aided only by benevolent contributions and without the interference o assistance of the Government. Without this restriction the responsibility of parents in rearing their offspring will b greatly diminished or annulled and a species of communis will be introduced. With regard to the method of teaching now in vogue I believe that there is too much cramming in our present system, that words are divorced from facts, and that names are taught without giving the child any know- ledge of the things indicated by them. Kindergarten methods of instruction seem to me to offer immense advan- tages over the time-honoured system of teaching childre from written characters and words. UNIVERSITY OF OXFORD : DEGREE DAYS.- Congregations will be held for the purpose of conferring degrees in Easter and Trinity terms on the following days viz. : Thursday, May 19th; Thursday, June 16th ; Thursday June 23rd ; and Saturday, July 9th.
https://openalex.org/W4205529760	https://zenodo.org/record/6643688/files/ATUAManual.pdf	English	null	Automated, Cost-effective, and Update-driven App Testing	ACM transactions on software engineering and methodology	2,022	cc-by	1,395	Prepared by Chanh Duc NGO Copyright: University of Luxembourg, SnT Centre for Security Reliability and Trust Version: V1 Prepared by Chanh Duc NGO Copyright: University of Luxembourg, SnT Centre for Security Reliability and Trust Version: V1 V1 ATUA User Manual Version: V1 1. Requirements - JDK 1.8+ is required for all the tools specified in the following sections. - Python 3 is required for Gator - Android SDK - Android 23+ emulator or device 1. Requirements - JDK 1.8+ is required for all the tools specified in the following sections. - Python 3 is required for Gator - Android SDK - Android 23+ emulator or device 2. Installation - ATUA can be run in a native environment on a local machin - To install, extract the toolset.zip - All the tools are provided with executables. - The ATUA’s toolset is comprised of: \|-- ATUA: contains ATUA's executable \|-- appdiff: contains Appdiff's executable \|-- extended-dm2-instrumenter: contains the extended version of DM2 instrumentation tool. \|-- extended-gator: contains Extended-Gator's executable - The ATUA’s toolset is comprised of: \|-- ATUA: contains ATUA's executable \|-- appdiff: contains Appdiff's executable \|-- extended-dm2-instrumenter: contains the extended version of DM2 instrumentation tool. \|-- extended-gator: contains Extended-Gator's executable \|-- template: templates for manual inputs and manual intents 3.Usage 3.Usage 3.1.2 Generate the EWTG App model This step concerns the generation of EWTG App model using an extended version of Gator. 3.1 Prepare the App under test ATUA needs the instrumented apk of the App under test, with a json providing instrumentation information, and the EWTG App model file. In addition, users can provide manual inputs and manual intents (a template for these inputs is provided in the template folder). Put the necessary files of the App under test into the folder Exploration.apksDir. The distribution comes with some case study Apps. The inputs files's formats are the following: ● Instrumented apk: [appPackage]_[v1]_[v2]-instrumented.apk. This file is generated by an extended version of Droidmate2 ● Instrumented info: [ appPackage]_[v1]_[v2].apk.json, contains the list of instructions and methods of the app under test. This file is generated with the instrumented file by an extended version of Droidmate2 ● Instrumented info: [ appPackage]_[v1]_[v2].apk.json, contains the list of instructions and methods of the app under test. This file is generated with the instrumented file by an extended version of Droidmate2 ● EWTG App model: [ appPackage]_[v1]_[v2]-AppModel.json. This file is generated by Extended-Gator. ● Manual input: [appPackage]_[v1]_[v2]-input.json. This file is expected to be filled with manual inputs to be used for testing an App. ● Manual intent: [appPackage]_[v1]_[v2]-intent.json. This file is expected to be filled with manual inputs to be used for testing an App intents. 2 ATUA User Manual ATUA User Manual Version: V1 Version: V1 3.1.1 Identify target methods and instructions This step concerns the identification of the methods modified and introduced by an updated version. This is accomplished by executing AppDiff. This step is not necessary to run ATUA with the provided case studies. AppDiff is provided as an executable. To run AppDiff, use the following command: java -jar appdiff.jar -o diff -a <android_sdk_jar> -j <output_folder> <path_to_oldVersionApk> <path_to_newVersionApk> A diff file in json format is produced and put in the <output_folder> 3.1.2 Generate the EWTG App model Gator. Before executing gator, put the app under test and the diff file in the same folder. For example: extended-gator/apk. If the diff file is not provided, the app under test will be analysed as all the methods are updated. To execute gator, use the following command from the extended-gator folder: ./gator a -p <Apk Path> -outputFile <EWTG App Model path> -worker 4 -appPackage <app package of the App under test> --timeout 36000 - worker: the number of processors which will be used - worker: the number of processors which will be used - worker: the number of processors which will be used - appPackage: the source code's package name. Some applications have their declared package name which does not match their source code's package name. This parameter allows gator to analyse these kinds of apps. - timeout: in second. The default timeout of Gator is 1 hour. This parameter - timeout: in second. The default timeout of Gator is 1 hour. This parameter could help to deal with large apps which require long processing time. 3.2.3 Instrument the app under test This step concerns the instrumentation of the app under test using an extended version of DM2 instrumentation project. - Firstly, put the apk file of the app under test and the diff file into the folder extended-dm2- instrumenter/apk. If the diff file is not provided, the app under test will be instrumented as all the methods are updated. - Firstly, put the apk file of the app under test and the diff file into the folder extended-dm2- instrumenter/apk. If the diff file is not provided, the app under test will be instrumented as all the methods are updated. - Run instrumentation from extended-dm2-instrumenter folder with the following command o useAppt2: some apps require appt2 to decompile and compile o useAppt2: some apps require appt2 to decompile and compile o packageName: the source code's package name. Some applications have their declared package name which does not match their source code's package name. This parameter allows the tool to instrument these kinds of apps. 3 ATUA User Manual ATUA User Manual Version: V1 - The output of this step is contained in the instrumentedApks folder, including the instrumented apk and the instrumented information. 3.3.2 Execute ATUA 3.3.2 Execute ATUA ATUA can be run from the command line. You can specify the configuration file to be used with the parameter “--Core-configPath”. The following command is run under the ATUA folder: ATUA can be run from the command line. You can specify the configuration file to be used with the parameter “--Core-configPath”. The following command is run under the ATUA folder: bin/ATUA --Core-configPath=./defaultConfig-ATUA.properties bin/ATUA --Core-configPath=./defaultConfig-ATUA.properties 3.3.3 Inspecting results 3.3 Run testing with ATUA 3.3.1 Prepare the testing configuration 1. Reuse defaultConfiguration-ATUA.property file or duplicate it for new purpose 2. In the configuration file, consider the following properties: ● Output.outputDir : the relative path of the output folder of the tool. If the folder does not exist, the tool will create the new folder. Attention: everytime ATUA is started, this folder will be cleaned. ex: Output.outputDir=./out60m/ATUA ● Exploration.apksDir: the relative path of the folder where the app under tests are located ex: Exploration.apksDir=./apk ● Exploration.deviceSerialNumber: the serial number of the device (got from "adb devices" command). Attention: please make sure that the device is available before executing the tool. ex: Exploration.deviceSerialNumber=emulator-5554 ● Selectors.timeLimit: testing time budget in minutes ex: Selectors.timeLimit ● Selectors.actionLimit: this is useful if you prefer the maximum number of actions as the overall budget ex: Selectors.actionLimit=1000 ● RegressionStrategy.budgetScale: scaleFactor to be used for each phase. The default value should be 1.0 for 1 hour testing budget. For five hours, we suggest using 2.0. 3.3.3 Inspecting results The result is stored in model/[appPackage] under the outputDir declared in the configuration file. ● ATUA-report.txt: contains the overall result in which you can find: ● ATUA-report.txt: contains the overall result in which you can find: ■ The number of statements (instructions) ■ The number of methods ■ The number of updated methods ■ The number of statements belong to the updated methods ■ The coverage ■ Some additional information including: ■ List of covered updated methods ■ The number of methods ■ Some additional information including: ■ List of covered updated methods 4 4 ATUA User Manual ATUA User Manual Version: V1 ■ List of uncovered update method ■ The coverage ratio after each phase ■ Unreachable windows ● coverage.txt: contains the list of instructions with the first covered timestamp. ● images: contains the screenshots of apps. ATUA takes a screenshot after every inpu being triggered. ● states: contains the captured GUITrees. ● trace<id>.csv: contains all the inputs generated. ■ List of uncovered update method ■ The coverage ratio after each phase ■ Unreachable windows age.txt: contains the list of instructions with the first covered timestamp. ● coverage.txt: contains the list of instructions with the first covered timestam ● images: contains the screenshots of apps. ATUA takes a screenshot after every input being triggered. ● images: contains the screenshots of apps. ATUA takes a screenshot after every input being triggered. ● states: contains the captured GUITrees. ● trace<id>.csv: contains all the inputs generated. 5 5
https://openalex.org/W2009014290	https://bmchealthservres.biomedcentral.com/counter/pdf/10.1186/1472-6963-9-71	English	null	Sharing clinical information across care settings: the birth of an integrated assessment system	BMC health services research	2,009	cc-by	8,288	BioMed Central BioMed Central Published: 29 April 2009 Published: 29 April 2009 Received: 31 March 2008 Accepted: 29 April 2009 ealth Services Research 2009, 9:71 doi:10.1186/1472-6963-9- This article is available from: http://www.biomedcentral.com/1472-6963/9/71 © 2009 Gray et al; licensee BioMed Central Ltd is article is available from: http://www.biomedcentral.com/147 © 2009 Gray et al; licensee BioMed Central Ltd. y ; This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: Population ageing, the emergence of chronic illness, and the shift away from institutional care challenge conventional approaches to assessment systems which traditionally are problem and setting specific. Methods: From 2002, the interRAI research collaborative undertook development of a suite of assessment tools to support assessment and care planning of persons with chronic illness, frailty, disability, or mental health problems across care settings. The suite constitutes an early example of a "third generation" assessment system. Results: The rationale and development strategy for the suite is described, together with a description of potential applications. To date, ten instruments comprise the suite, each comprising "core" items shared among the majority of instruments and "optional" items that are specific to particular care settings or situations. Conclusion: This comprehensive suite offers the opportunity for integrated multi-domain assessment, enabling electronic clinical records, data transfer, ease of interpretation and streamlined training. Open A Correspondence Sharing clinical information across care settings: the birth of an integrated assessment system Leonard C Gray1, Katherine Berg2,3, Brant E Fries4,5, Jean-Claude Henrard6 John P Hirdes7,8, Knight Steel9 and John N Morris10 Address: 1The University of Queensland. c/- Academic Unit in Geriatric Medicine, University Department of Medicine, Princess Alexandra Hospital, Brisbane, Australia, 2University of Toronto, Toronto, Canada, 3University of Waterloo, Waterloo, Canada, 4Institute of Gerontology, School of Medicine, University of Michigan, Ann Arbor, USA, 5Health Systems Research, Veterans Administration Healthcare System, Ann Arbor, Michigan, USA, 6Versaille University, Paris-Oust Medical School, Paris, France, 7Department of Health Studies and Gerontology, University of Waterloo, Waterloo, Canada, 8Homewood Research Institute, Waterloo, Ontario, Canada, 9Hackensack University Medical Center, New Jersey, USA and 10Institute for Aging Research, and Alfred A and Gilda Slifka Chair in Social Gerontological Research, Hebrew Senior Life, Boston, Massachusetts, USA Email: Leonard C Gray - len.gray@uq.edu.au; Katherine Berg - katherine.berg@utoronto.ca; Brant E Fries - bfries@umich.edu; Jean- Claude Henrard - jean-claude.henrard@spr.aphp.fr; John P Hirdes - hirdes@uwaterloo.ca; Knight Steel - ksteel@humed.com; John N Morris - jnm@mail.hrca.harvard.edu * Corresponding author Received: 31 March 2008 Accepted: 29 April 2009 Open A Correspondence Sharing clinical information across care settings: the birth of an integrated assessment system Leonard C Gray*1 Katherine Berg2,3 Brant E Fries4,5 Jean Claude Henrard6 Open Access http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 BMC Health Services Research 2009, 9:71 tings. By focusing on a limited set of care prerogatives, health care systems often fail to respond adequately to the complex interactions of factors influencing the lives of persons who turn to formal care systems for assistance. being the site of care for multiple individuals with quite different needs who therefore require strikingly different services. Nonetheless, although certain diseases, levels of acuity and functional deficits may be more common in one location than another and may be required to justify care in a particular site, the specific needs of each individ- ual must be addressed appropriately regardless of that per- son's location. To both improve outcomes and maximize the function of those served, essential health-related information must be transmitted from site to site in a timely manner. The elec- tronic medical record can make such transmission across sites of care easier. However, more than ever, the success- ful introduction of electronic medical records will depend on the availability of high quality, standardized, clinically relevant data that carry the same meaning independent of the location of care. Moreover, the information must be accessible and useful to diverse stakeholders responding to different issues at various levels of the health care sys- tem. Availability of clinical information is one of the key ingredients of successful strategies to better manage chronic illness [4]. Older adults not only use more care, but they receive care in a host of sites, such as hospitals, long term care facilities and even the home. Further, because many older individ- uals have multiple chronic diseases, the conditions of a particular person that are being addressed in one location often are similar if not identical to those in the site to which the person is next transferred. Rather than targeted to a new disease, the care in the second site usually must address changes in acuity and perhaps the interaction of one or more chronic diseases with a newly discovered condition. Similarly, the transition to inpatient psychiatry from community mental health services for a middle aged adult with schizophrenia is more likely to be driven by a change in the severity of symptoms than the onset of a new type of mental illness. Thus, a crucial ingredient of future multi-sector care deliv- ery systems will be a common language of clinical descrip- tors. Page 2 of 10 (page number not for citation purposes) http://www.biomedcentral.com/1472-6963/9/71 This language must be broad in its conceptualization, detailed when necessary, and capable of being interfaced, where possible, with emerging com- puter-based information technologies. In industrialized countries, there has been a substantial shift away from institutional models of care of chronic ill- ness, particularly in mental health and disability services. Many governments are now adopting similar approaches to care of older persons. These transformations increase the complexity of the service delivery framework, with multiple service providers often delivering care to individ- uals, and more frequent changes of care setting occurring, particularly between community and facility settings. Integration and coordination have become central issues in the care of older people, and a variety of demonstration programs have been established in response. [2,3] The concept of third generation assessment instruments The concept of third generation assessment instruments The concept of third generation assessment instruments Many clinical and care services adopt structured approaches to assessment. Typically, these consist of items developed by the organization itself or a panel of assess- ment tools that attend to aspects of assessment (e.g., the Mini-Mental State Examination for dementia screening, or the Barthel Index for activities of daily living profiling). These systems can be characterized as "first generation" assessment instruments. Their strength is their focus on a specific issue or problem, the establishment of discrete measurement rules, and field testing of the tools for relia- bility, validity, and utility within a clinical trial environ- ment. Their weakness is the lack of proven utility across different care settings, the focused nature of their assess- ment, and the difficulty of "cobbling together" a series of what are often lengthy single-purpose tools into a usable, coherent overall assessment battery. These changes demand radical re-rethinking of the organ- ization of clinical information systems that typically have been designed to support single service providers in one setting. Traditional clinical systems tend to focus on a lim- ited set of problems, often reflecting the views of care pro- fessionals and funding agencies. Complex, multi- dimensional views of the person seldom come to the fore in such systems. For those who see health and social care from such a perspective, the number and types of mean- ingful problems is limited, as is the scope of the assess- ment methodologies and the care they are provided. Second generation assessment instruments aim to cross many clinical domains and have applicability in many settings. They also provide a greater opportunity to inter- pret assessment information systematically to assist in treatment and care planning. This type of instrument requires a broad consensus on the multiplicity of domains to be measured. Background tems of each nation. Thus a country's health care structure may stipulate what services are reimbursed at each level of care and thereby effectively preclude their being provided in other locations. Also the availability of informal sup- port systems or lack of them may result in a given location g The purpose of health care is to provide person-specific rather than site-specific care [1]. With rare exceptions, the site of care is determined by economic considerations and by the structure and policies of the health and welfare sys- Page 1 of 10 (page number not for citation purposes) Page 1 of 10 (page number not for citation purposes) The concept of third generation assessment instruments As with first generation tools, the items must be as relevant as possible to the underlying domains; the only difference is that there is a need for a purposeful trade-off between the number of operational items for any With this as backdrop, there is a need for assessment sys- tems and service provision that transcend care settings, enabling the identification of and response to complex needs with person-specific care planning information that can flow with individuals as they move across care set- Page 2 of 10 (page number not for citation purposes) Page 2 of 10 (page number not for citation purposes) BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 adopted the RAI-MDS on a widespread basis include Can- ada, Finland, Estonia, and Iceland. A series of "case stud- ies" have been published by the Milbank Memorial Fund [10]. Intervention studies suggest that implementation of instruments may influence outcomes [11]. one domain and the overall length of the assessment tool. In a second generation tool, individual items are con- structed to record focused information about the individ- ual and it is the assembly of these items into a meaningful set of purposeful domains that form the backbone of a comprehensive assessment schedule. A key attribute of such instruments is the attention to the primary purpose of care planning, i.e., to provide the user with information targeting specific medical, functional, and social prob- lems that need to be addressed and further guidance through care planning protocols. The instrument must then be subjected to extensive field testing to ensure relia- bility of items and validity of the summary measures that are based on the items. Such a tool has the advantage of providing consistency in item structure; coverage of all rel- evant care domains; information about the person's needs to guide care planning and the ability to build a robust set of outcome products such as case-mix groupings, quality indicators and eligibility determinants [5]. In 1994, a home care version was created – the RAI-HC – which has also enjoyed considerable implementation in several countries [12]. Subsequently, instruments were developed for acute and post-acute care, palliative care, hospital and community mental health, physical and intellectual disability and assisted living. interRAI instruments interRAI instruments are administered by trained asses- sors who interact with the person, their caregiver(s) and staff (particularly in institutional settings). They also review all available records. It is only after consulting all of these sources of information that "observations" are determined, based on their best judgment if the informa- tion is conflicting. For example, in order to determine whether an elderly woman prepares her own meals or per- forms her own housekeeping in a home care service set- ting, the assessor will consult the person, her care-giver(s), community service providers, and any available records. The exception is a small number of items that are specifi- cally addressed to the person being assessed, such as aspects of mood and self- reported perception of health. Where there is evidence of cognitive or communication impairment, there will be increased reliance on informa- tion provided by care-givers and staff. This approach is complex and requires good judgment on the part of the assessor, but repeatedly has been shown to be reliable. Interviews with persons are therefore conversational in style, and semi-structured. Third generation instruments extend the concept of sec- ond generation tools to multiple care settings. They pro- vide assessment strategies that transcend boundaries between care settings. The focus is on the changing strengths, preferences and needs of the person, rather than the sector which happens to be providing services at a sin- gle point in time. In 2002, the interRAI consortium took up the challenge to refine its array of second-generation assessment tools into a truly integrated third-generation system. The remainder of this paper describes the approach taken and the out- comes of the process to date. The concept of third generation assessment instruments The initial focus of interRAI on care of older persons has since been broad- ened to address the needs of adults of all ages with com- plex and disabling physical and mental illnesses. The interRAI consortium interRAI is a not-for-profit research consortium of about 50 clinicians, researchers and health administrators from 25 countries. It was established in 1992 with a collective vision that "the assembly of accurate clinical information in a common format within and across services sectors and countries enhances both the well-being of frail per- sons and the efficient and equitable distribution of resources" [6]. Once the assessment is completed, scales summarizing major domains (e.g., depression, cognition, and physical function) are calculated. They are not explicitly adminis- tered but are computed from the overall observation. The burden of the assessment process is thus minimized. [Copies of interRAI instruments may be obtained by con- tacting interRAI at http://www.interrai.org. Instruments are copyright, however they are made freely available to non-commercial organizations.] Initially, development efforts focused on long-term resi- dential care, with the Resident Assessment Instrument – Minimum Data Set (RAI-MDS) and its associated case-mix application (Resource Utilization Groups – RUG-III [7]) enjoying wide uptake in nursing homes in North America and Europe. An early version of the instrument was man- dated in all Medicare and Medicaid funded nursing homes in the USA in 1990, and has been associated with measurable improvements to the standard of care, partic- ularly when quality indicators derived from the instru- ment were introduced [8,9]. Other nations that have since Initially, development efforts focused on long-term resi- dential care, with the Resident Assessment Instrument – Minimum Data Set (RAI-MDS) and its associated case-mix application (Resource Utilization Groups – RUG-III [7]) enjoying wide uptake in nursing homes in North America and Europe. An early version of the instrument was man- dated in all Medicare and Medicaid funded nursing homes in the USA in 1990, and has been associated with measurable improvements to the standard of care, partic- ularly when quality indicators derived from the instru- ment were introduced [8,9]. Other nations that have since interRAI instruments have the following design features: interRAI instruments have the following design features: • A fully developed instrument comprises a data set (for clinical data collection), a training manual and a set of algorithms that generate "Clinical Assessment Protocols" Page 3 of 10 (page number not for citation purposes) BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 (CAPs), scales (including screeners and severity meas- ures), case-mix measures and quality indicators. (CAPs), scales (including screeners and severity meas- ures), case-mix measures and quality indicators. of health professionals (typically 2 or 3 days), high levels of reliability and validity are assured. of health professionals (typically 2 or 3 days), high levels of reliability and validity are assured. • Instruments are designed for application by health pro- fessionals trained in the interRAI method with the support of a training manual. • Instruments are designed for application by health pro- fessionals trained in the interRAI method with the support of a training manual. • There is a development path for each instrument. Revi- sions (or upgrades) are produced as clinical practice standards develop. The most mature instrument (for long term residential care of older persons) is now in its fourth version and the home care instrument in its third version. • The instruments primarily inform the clinical assess- ment and care planning processes – administrative utili- ties such as planning data, case-mix tools and quality indicators are by-products. Thus, interRAI instruments are designed to produce obser- vations and outputs primarily to assist clinicians in assess- ment and care planning, but have the added advantage of providing an array of purposefully designed information for a wide range of stakeholders. (Figure 1). • The instruments have sound psychometric properties, established through extensive field testing and revision of poorly performing items. With moderate levels of training Schematic representation of an interRAI instrument Figure 1 Schematic representation of an interRAI instrument. Schematic representation of an interRAI instrument Figure 1 Schematic representation of an interRAI instrument Figure 1 Schematic representation of an interRAI instrument. Schematic representation of an interRAI instrument Figure 1 Schematic representation of an interRAI instrument. Page 4 of 10 (page number not for citation purposes) BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 There have been many spin-off applications of data from interRAI instruments, including determination of eligibil- ity for services; tracking of individuals' progress over time; structures for resource allocation both within organiza- tions and across nations; rationalizing of services and sys- tems; and projection of need and associated costs [6]. It has been used for cross-national comparisons of needs [13], which contributes to a better understanding of response options related to those needs. More often, the transfer is associated with a change in sta- tus. For example, when there is an admission to hospital there may be changes in functional status associated with a new disabling illness. Similarly, when a person is admit- ted to a long term care facility or psychiatric hospital from the community, there may be changes in mood and behavior. In this circumstance, prior information around the person's status would provide valuable insights regarding his/her current needs. If a person has a progres- sive or fluctuating condition, and is moving between care settings, monitoring of change over time would assist cli- nicians to assess stability and rate of progression. Until 2001, different instruments were designed by differ- ent committees comprised of domain experts and meas- urement scientists from within interRAI and collaborating organizations. In general, the starting point for new instruments was a thorough review of the key issues that were relevant to persons within the care setting for which the instrument was completed. Extensive lists of issues were drawn up, subjected to extensive internal and exter- nal review, and only then did the interRAI team begin to "create" the first draft of the instrument. Wherever possi- ble, each team endeavored to use applicable key measures and relevant material from earlier interRAI assessment tools. Thus, when interRAI moved to the RAI-HC, the home care instrument drew heavily on items contained in the nursing home instrument. Where necessary, commit- tees modified items to best suit their care setting. New items were developed, without necessarily referring to items measuring similar concepts in other instruments. 4) There were perceived administrative advantages in addition to those derived from improvements in the effi- ciency of clinical assessment procedures apparent from the above. In particular, design and construction of soft- ware systems to support multi-sector assessment and care planning would be simplified and the availability of com- mon assessment items with common definitions would enable multiple instruments (and new ones) to be built quickly. Recognition of this inconsistency, coupled with an emerg- ing vision of an integrated health information system – one that permitted seamless tracking of persons in multi- ple service settings – led to the establishment of a process to create the third-generation interRAI suite of instru- ments. As a result, interRAI developed a set of instruments with a similar look and feel, but which in the details did not con- sistently measure the identical phenomena in the identi- cal way. This was not necessarily a problem if one focused only on care in a single setting; however, it fell short of the full potential of the family of instruments to function as a system linking multiple sectors together. 2) Consistent recording of information across care set- tings might assist clinicians in other ways: Familiarity with the language, definitions and interpretation of an instru- ment utilized in the health professionals "home" care set- ting would facilitate interpretation of a companion instrument performed in another setting. For example, if a measure of ADL dependency is constructed and scored in the same manner in all settings, interpretation of the level of dependency will be simplified. Thus, when a per- son is discharged from a post-acute service to a home care program, the admitting assessor would quickly be able to appraise the person's needs even before consulting him/ her directly. 2) Consistent recording of information across care set- tings might assist clinicians in other ways: Familiarity with the language, definitions and interpretation of an instru- ment utilized in the health professionals "home" care set- ting would facilitate interpretation of a companion instrument performed in another setting. For example, if a measure of ADL dependency is constructed and scored in the same manner in all settings, interpretation of the level of dependency will be simplified. Thus, when a per- son is discharged from a post-acute service to a home care program, the admitting assessor would quickly be able to appraise the person's needs even before consulting him/ her directly. 3) Training of assessors would be simplified. A nurse trained in the use of an instrument in one sector could quickly learn to use a companion instrument in another. Many health professionals work across sectors either con- currently or over the course of their career. 4) There were perceived administrative advantages in addition to those derived from improvements in the effi- ciency of clinical assessment procedures apparent from the above. In particular, design and construction of soft- ware systems to support multi-sector assessment and care planning would be simplified and the availability of com- mon assessment items with common definitions would enable multiple instruments (and new ones) to be built quickly. Page 5 of 10 (page number not for citation purposes) The development process • Commonly-used, standardized interRAI scales (e.g., the Cognitive Performance Scale, and ADL scales [19,20]) In 2001, interRAI established a multinational and multi- disciplinary working group of fifteen researchers to create an integrated suite of assessment instruments. During the five year development and testing process the group met for multi-day meetings on thirteen occasions. An estimate of the contributed cost of developing this system already exceeds US$2.5 million. The group continues to meet to support further development of the suite. Items that served at least one of these purposes were given priority to be selected as core items. Items that served at least one of these purposes were given priority to be selected as core items. Committee members solicited and received extensive international feedback from clinicians, researchers and organizations around the world using one or more inter- RAI instruments. Then items were reviewed and, if neces- sary, the wording harmonized to ensure consistency across all instruments. Rules for item design were created, and rigorously applied to each item: The initial challenge to the group was to create a truly inte- grated suite of assessment tools for long-term institutional care, post-acute care, home care, assisted living, in-patient and out-patient mental health, palliative care and inde- pendent living settings, and one that would be applicable worldwide. The first step was to identify a set of domains and associated operational items that were considered mandatory for each new instrument – "core items." The second task was to create an optional set of domains and associated items that would appear on several, but not all, instruments in the suite. Finally, there would be instru- ment specific domains and items that were applicable on only one or two settings. [Examples of core items are cog- nitive skills for daily decision -making and mobility. Examples of optional items are certain aspects of memory function or pain. Items related to suicide and dying are specific to mental health and palliative care instruments respectively.] • interRAI items are phrased to give clear definition, and they require observations of behaviors or per- formance over a specific time frame (usually 3 days for the new suite). Item descriptions typically include examples and exclusions to increase the precision of definitions. • The assessment process follows a usual clinical inter- view, assimilating information from multiple sources. The instrument suite concept 5) Comparison of case complexity of persons in different care settings would be facilitated. For example, adminis- trators concerned with the conundrum of relative costs of community and institutional care could compare caseloads against a wide array of measures. interRAI tools have proven extremely successful in comparative analyses of caseloads across similar care settings in different juris- dictions, at local, national and international levels [13,14]. This capacity to compare caseloads would serve to improve equity of access to services. The concept of a "suite" of compatible assessment tools arose from the following six considerations: 1) People with chronic illness and disability often move between care settings. There is usually a need to transfer clinical information about the person. If, at the time of transfer, the person's condition or circumstances are sta- ble, the information from the most recent assessment would be current, with the potential to reduce the assess- ment effort by the receiving agency. In some instances it might eliminate the need completely. Page 5 of 10 (page number not for citation purposes) Page 5 of 10 (page number not for citation purposes) BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 National Quality Framework approved quality meas- ures posted on the Centers for Medicare and Medicaid Services website [15]. Quality indicators have also been developed for home care [16], post-acute care [17]), and mental health; [18] 6) Finally, if instruments are designed to minimize the effect of cultural differences, they provide a powerful capacity to compare the utilization and impact of differ- ent health and social systems across nations. Page 6 of 10 (page number not for citation purposes) http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 BMC Health Services Research 2009, 9:71 Extensive international field testing was undertaken. The first five instruments completed were studied: long term care; home care; post-acute care; palliative care and inpa- tient mental health. Field testing focused on item response rates; inter-rater reliability; and convergent validity of scales within each instrument, typically consid- ering frequently used scales as the benchmark standard. The results are the subject of a separate publication[21], and the reliabilities were closely comparable to previously reported values [22-24]. individuals who are likely to appear in more than one set- ting are also coordinated. For example, the majority (68%) of items in the interRAI AC are contained within the interRAI HC; and the interRAI MH contains 84% of the items in the interRAI CMH. Because of frequent move- ment of individuals between hospital and community set- tings, this overlap is expected to facilitate a common understanding of the person's needs, and to thus improve continuity of care. Interpretation of clinical observations Subsequently, several other instruments have been added to the suite, using a similar development approach. Exam- ples include new instruments for acute care [25], commu- nity mental health, intellectual disability, and assisted living. In addition, new screeners for psychiatry, home care, and emergency department settings were created to be compatible with the full assessments in the suite. In addition to the "minimum data sets" of clinical obser- vations, interRAI instruments comprise manuals to sup- port training and a series of algorithms that "interpret" the clinical findings. The algorithms generate scales that provide severity meas- ures (e.g., the extent of ADL dependency) or diagnostic screeners (e.g., whether a person has dementia). A group of scales were developed for previous generations of instruments [19,20,26-29]. These have been adapted to the new suite, for use across the range of care domains. Numerous scales are currently available encompassing cognition, communication, mood (depression), instru- mental and personal ADL, pain and health stability. The concurrent validity of several scales (against comparable scales in widespread use, such as the Mini-Mental State Examination and the Barthel Index) was re-tested in the initial international field work, with good results. The findings will be reported in a future paper. Further scale development and validation is anticipated as the suite comes into wider use. The development process Clinicians use their professional judgment to appraise diverse information and to assign the most appropri- ate code to the form. • The psychometric and distributional properties of items are scrutinized carefully to ensure reliability, validity and clinical relevance. This task was undertaken by compiling a comprehensive inventory of all existing interRAI items. The inventory was reviewed to identify concepts and items in each instru- ment – some were shared, others were unique. Among shared items, variations were identified. • Items are not survey questions that are asked pre- cisely the same way and in the same sequence. Rather, the response categories coded by clinicians are the focus of standardization. Next, the function of items within specific instruments was catalogued. In addition to describing some aspect of a person, item functions included contributions to: • To facilitate applicability to multiple settings the suite refers to "persons" rather than patients, clients or residents. • Clinical Assessment Protocols (CAPs), which are algorithms that suggest care planning actions; • Case-mix classification systems on which reimburse- ment formulas were based (e.g., RUG-III for nursing homes, RUG-III/HC for home care, and SCIPP for inpatient psychiatric care); New instruments were then assembled for each care set- ting. Core items were considered mandatory and only excluded in exceptional circumstances, where they were clearly not appropriate for a particular setting. Optional items were then considered, and included if considered relevant to the setting. Finally, setting (or instrument) spe- cific items were added. • Quality indicator systems. In the nursing home RAI this includes the quality measures created by the Zim- merman group, the Morris Mega-QI group, and the Page 6 of 10 (page number not for citation purposes) Page 6 of 10 (page number not for citation purposes) http://www.biomedcentral.com/1472-6963/9/71 The outcomes At the time of writing, ten instruments were complete (Table 1). Several additional instruments are planned for the immediate future. The instrument data sets range in size from 96 (interRAI AC) to 320 (interRAI CMH). As a general rule, where there was a prior interRAI instrument, the version created for the new suite was notably shorter. The instruments in the new suite share a large proportion of items – a result of the design strategy. The core items represent over three quar- ters of most instruments, providing a "backbone" of criti- cal assessment information. Instruments applying to Table 1: The interRAI suite of assessment instruments Instrument Item count Target population Long term care facility (interRAI LTCF) 257 Residents of nursing homes or chronic hospitals Assisted living (interRAI AL) 262 Assisted living facilities where residents have light care needs Acute care (interRAI AC) 96 Frail older patients in acute care hospitals Post-acute care (interRAI PAC) 214 Rehabilitation and other post-acute inpatients Home care (interRAI HC) 253 Community based care Community health assessment (interRAI CHA) 135 Community settings with anticipated light care needs Palliative care (interRAI PC) 194 Palliative care in community and institutional settings Mental health (interRAI MH) 304 Mental health inpatients Community mental health (interRAI CMH) 320 Community mental health services Intellectual disability (interRAI ID). 287 Persons with intellectual disability in community and facility settings Table 1: The interRAI suite of assessment instruments BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 Clinical assessment protocols (CAPs) are an integral fea- ture of each individual assessment instrument in the suite. CAPs interpret clinical observations in order to identify opportunities to influence clinical and other outcomes. They perform a combination of functions including prob- lem definition (e.g., the person has incontinence) and indicating an opportunity to intervene (e.g., prevention of falls or pressure ulcer, or improvement of ADL function). The method of development utilized by interRAI to develop CAPs comprised several tasks: review of the rele- vant scientific literature (e.g. strategies that prevent falls in an at risk population); analysis of existing extensive inter- RAI data sets to identify sub-populations with adverse (or in some cases positive) outcomes across multiple assess- ment periods; and expert opinion provided by interRAI members and their associates. for clinicians, administrators and researchers. However, implementation of such systems may meet substantial challenges. Necessarily, implementations will be large scale. Administrative tools Reluctance to share instrumentation is also evident in the clinical milieu, particularly in multi-disciplinary settings. Each professional group uses its own instrument set to appraise aspects such as cognition, ADL and mood. There is often a division of labor in which each profession attends to aspects of the problem – medical staff to cogni- tion, occupational therapists to functional activities, and nurses to pressure ulcer prevention. The introduction of a "shared" clinical dataset can be perceived to compromise the quality of instrumentation and may threaten profes- sional autonomy. interRAI instruments have proven particularly useful in a variety of administrative areas. Quality indicators devel- oped for nursing homes have been associated with improvement in care standards [9]. Case-mix tools in long term residential care of older persons and community care are used in several nations as a basis for funding. The extensive clinical information derived from interRAI data- sets has provided a powerful capacity to compare caseload complexity and service responses between facilities, regions and nations [7,30]. interRAI is currently updating all of these relevant toolsets for application to the inte- grated suite. Conversely, the use of a shared electronic dataset may present an opportunity to improve productivity through reduction in duplication of data collection, particularly if this information is linked to a wider system that brings previous data to the current setting, and offers an oppor- tunity to pass on information efficiently to the subsequent care setting. The outcomes The establishment of systems across settings is likely to require significant investment by governments. Specific services are often separately administered by different levels of government – national, state, and regional. Even within one level of government, administrative "silos" can develop which attend to each service type (e.g., hospitals and community care). To further complicate matters, the health and social service systems are usually organized in their own administrative silos. Finally, many elements of service systems are often provided by private agencies with varying levels of independence from government admin- istration. The application of a single integrated clinical information system to such a complex jigsaw of services seems formidable. In the new suite, the emphasis has been shifted towards identification of problems where there is evidence that the outcome can be influenced. To date, over 40 CAPs have been developed, ranging from clinically oriented prob- lems such as pressure ulcer and pain, to social issues such as abusive behavior and social relationships. A CAP con- sists of a computer algorithm which identifies persons to whom the protocol applies, and a set of guidelines which assist clinicians to mount an appropriate response. The guideline component is authored by an international panel of relevant domain experts. The CAPs may be used in their own right as a form of clinical guideline, or they may be used in conjunction with existing guidelines. However, the interRAI CAPs have the advantage of being intimately linked to the assessment process. There are nations and service arrangements where full implementation is potentially feasible. However, it is more likely that an incremental approach will be success- ful. Administrations operating two or more related serv- ices might apply two or three instruments (e.g., acute and post-acute care; community and institutional mental health). Services with relationships to the organizations that implement instruments may see opportunities to improve inter-operability by implementing other instru- ments from the suite. Ultimately, the choice not to share instrumentation may place an organization at considera- ble disadvantage. This effect is likely to be particularly pronounced with a fully-integrated suite. Conclusion h The interRAI suite of instruments represents, to our knowledge, the first major effort to provide an integrated health information system with the potential to provide person-centered information that transcends care settings. It offers opportunities to improve continuity of care, from both efficiency and quality of care perspectives. Training of assessors in the interpretation and application of infor- mation drawn from a variety of service settings would be simplified. Administrators and planners would be able to compare caseloads and outcomes across settings. When used to its full potential the interRAI suite can substan- tially enhance the delivery of health care as a system responding effectively to the needs of vulnerable popula- tions over their life course. John N. Morris. PhD. Co-Director, Institute for Aging Research, and Alfred A and Gilda Slifka Chair in Social Gerontological Research, Hebrew Senior Life, Boston, Massachusetts, USA. The authors declare that they have no competing interests. y y ( ) 3. Leichsenring K, Alaszewski AM: Providing integrated health and social care for older persons: a European overview of issues at stake. In Providing integrated health and social care for older persons: a European overview of issues at stake Farnham: Ashgate; 2004. 3. Leichsenring K, Alaszewski AM: Providing integrated health and social care for older persons: a European overview of issues at stake. In Providing integrated health and social care for older persons: a European overview of issues at stake Farnham: Ashgate; 2004. 4. Wagner EH, Austin BT, Von Korff M: Organizing care for patients with chronic illness. Milbank Q 1996, 74(4):511-544. 5. Hirdes JP, Fries BE, Morris JN, Steel K, Mor V, Frijters D, LaBine S, Schalm C, Stones MJ, Teare G, et al.: Integrated health informa- tion systems based on the RAI/MDS series of instruments. Healthc Manage Forum 1999, 12(4):30-40. Authors' information Leonard C. Gray. MD. PhD. Professor in Geriatric Medi- cine, The University of Queensland, c/- Academic Unit in Geriatric Medicine, University Dept of Medicine interRAI systems are currently involved in research aimed at improving inter-operability of clinical informations sytems, such as the HL7 standards and Snomed CT clinical nomenclature systems. interRAI systems are of particular interest to this area of technical development because of their widespread use, good psychometric properties and holistic approach to data collection. The use of the same items across clinical settings may simplify some of the translational issues when clinical concepts (such as mobility or memory) are shared. Katherine Berg. PhD. PT. Department Chair of Physical Therapy, University of Toronto, Canada, and Adjunct Pro- fessor, University of Waterloo, Canada. Katherine Berg. PhD. PT. Department Chair of Physical Therapy, University of Toronto, Canada, and Adjunct Pro- fessor, University of Waterloo, Canada. Brant E. Fries. PhD. Professor, Health Management and Policy; Research Professor, Institute of Gerontology, School of Medicine, University of Michigan; and Chief, Health Systems Research, Veterans Administration Healthcare System, Ann Arbor, Michigan, USA. Jean-Claude Henrard. MD. Professor, Research Unit Health and Ageing, Versaille University, Paris-Oust Medi- cal School, Paris, France. The distribution of personal information across care set- tings, notably when the care is provided by different agen- cies, requires careful attention to privacy issues. Protocols are required to ensure that individuals are comfortable with the sharing of information among their caregivers. However, this is a universal concern which is not specific to the interRAI suite. John P. Hirdes. PhD. Professor, Department of Health Studies and Gerontology, University of Waterloo and Sci- entific Director, Homewood Research Institute, Waterloo, Ontario, Canada. Knight Steel. MD. Chief, Geriatric Medicine, Hackensack University Medical Center, New Jersey, USA. References 1. Steel K, Sherwood S, Ribbe MW: The future: a person-specific standardized assessment instrument. Age Ageing 1997, 26(Suppl 2):83-85. ( pp ) 2. Johri M, Beland F, Bergman H: International experiments in inte- grated care for the elderly: a synthesis of the evidence. Int J Geriatr Psychiatry 2003, 18(3):222-235. ( pp ) 2. Johri M, Beland F, Bergman H: International experiments in inte- grated care for the elderly: a synthesis of the evidence. Int J Geriatr Psychiatry 2003, 18(3):222-235. Acknowledgements The authors acknowledge the contributions made by members of the inter- RAI Instrument and Systems Development (ISD) Committee. The authors acknowledge the contributions made by members of the inter- RAI Instrument and Systems Development (ISD) Committee. The authors acknowledge the contributions made by members of the inter- RAI Instrument and Systems Development (ISD) Committee. RAI Instrument and Systems Development (ISD) Committee. Implementation issues The instrument data sets were released during 2005 and have begun to come into active use in several countries. Several multi-setting demonstrations are being estab- lished across the world [31]. The availability of an integrated multi-setting clinical information system provides a wide array of opportunities Page 8 of 10 (page number not for citation purposes) http://www.biomedcentral.com/1472-6963/9/71 BMC Health Services Research 2009, 9:71 actively contributed to the research program and partici- pated in the many research forums described in the man- uscript. All authors read and approved the final manuscript. interRAI instruments are intimately dependent on com- puterization, since an integral feature is a suite of complex algorithms that generate scales, CAPs and numerous other administrative by-products. Thus, a major challenge to implementation is access to computers and the need for a degree of computer literacy. Many service settings are not yet ready for this level of sophistication. However, as the benefits of such systems grow, the case for investment in these capabilities becomes increasingly powerful. Competing interests p g The authors declare that they have no competing interests. Page 9 of 10 (page number not for citation purposes) Pre-publication history J Gerontol A Biol Sci Med Sci 1999, 54(11):M546-553. 21. Hirdes JP, Ljunggren G, Morris JN, Frijters D, Finne-Soveri H, Gray LC, Bjorkgren M, Gilgen R: Reliability of the interRAI Suite of Assessment Instruments: A 12-Country Study of an Inte- grated Health Information System. BMC Health Services 2008, 8:277. 22. Sgadari A, Morris JN, Fries BE, Ljunggren G, Jonsson PV, DuPaquier JN, Schroll M: Efforts to establish the reliability of the Resident Assessment Instrument. Age Ageing 1997, 26(Suppl 2):27-30. g g g ( pp ) 23. Hawes C, Morris JN, Phillips CD, Mor V, Fries BE, Nonemaker S: Reliability estimates for the Minimum Data Set for nursing home resident assessment and care screening (MDS). Geron- tologist 1995, 35(2):172-178. Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Publish with BioMed Central and every scientist can read your work free of charge 24. Hirdes JP, Smith TF, Rabinowitz T, Yamauchi K, Perez E, Telegdi NC, Prendergast P, Morris JN, Ikegami N, Phillips CD, et al.: The resident assessment instrument-mental health (RAI-MH): Inter-rater reliability and convergent validity. J Behav Health Ser R 2003, 29(4):419-432. ( ) 25. Gray L, Bernabei R, Berg K, Finne-Soveri H, Fries B, Hirdes J, Jonsson P, Morris J, Steel K, Arino-Blasco S: Standardizing assessment of the elderly in acute care: the interRAI Acute Care instru- ment. Journal of the American Geriatric Society 2008, 56(3):536-41. J y ( ) 26. Pre-publication history [ p g p ] 11. Bernabei R, Landi F, Gambassi G, Sgadari A, Zuccala G, Mor V, Ruben- stein LZ, Carbonin P: Randomised trial of impact of model of integrated care and case management for older people living in the community. Bmj 1998, 316(7141):1348-1351. 12 M JN F BE S l K Ik N B b R C GI The pre-publication history for this paper can be accessed here: The pre-publication history for this paper can be accessed here: The pre-publication history for this paper can be accessed here: 12. Morris JN, Fries BE, Steel K, Ikegami N, Bernabei R, Carpenter GI, Gilgen R, Hirdes JP, Topinkova E: Comprehensive clinical assess- ment in community setting: applicability of the MDS-HC. Journal of the American Geriatrics Society 1997, 45(8):1017-1024. http://www.biomedcentral.com/1472-6963/9/71/prepub http://www.biomedcentral.com/1472-6963/9/71/prepub 13. Carpenter I, Gambassi G, Topinkova E, Schroll M, Finne-Soveri H, Henrard JC, Garms-Homolova V, Jonsson P, Frijters D, Ljunggren G, et al.: Community care in Europe. The Aged in Home Care project (AdHOC). Aging Clin Exp Res 2004, 16(4):259-269. 14 F BE S h ll M H C G l R J PV P k P p j ( ) g g p ( ) 14. Fries BE, Schroll M, Hawes C, Gilgen R, Jonsson PV, Park P: Approaching cross-national comparisons of nursing home residents. Age Ageing 1997, 26(Suppl 2):13-18. g g g ( pp ) 15. Zimmerman DR, Karon SL, Arling G, Clark BR, Collins T, Ross R, Sainfort F: Development and testing of nursing home quality indicators. Health Care Financ Rev 1995, 16(4):107-127. 16. Hirdes JP, Fries BE, Morris JN, Ikegami N, Zimmerman D, Dalby DM, Aliaga P, Hammer S, Jones R: Home care quality indicators (HCQIs) based on the MDS-HC. Gerontologist 2004, 44(5):665-679. ( ) 17. Morris JN, Jones RN, Fries BE, Hirdes JP: Convergent validity of minimum data set-based performance quality indicators in postacute care settings. Am J Med Qual 2004, 19(6):242-247. 18. Hirdes JP, Marhaba M, Smith TF, Clyburn L, Mitchell L, Lemick RA, Telegdi NC, Perez E, Prendergast P, Rabinowitz T, et al.: Develop- ment of the Resident Assessment Instrument – Mental Health (RAI- MH). Hosp Q 2000, 4(2):44-51. 19. Morris JN, Fries BE, Mehr DR, Hawes C, Phillips C, Mor V, Lipsitz LA: MDS Cognitive Performance Scale. J Gerontol 1994, 49(4):M174-182. ( ) 20. Morris JN, Fries BE, Morris SA: Scaling ADLs within the MDS. http://www.biomedcentral.com/1472-6963/9/71 http://www.biomedcentral.com/1472-6963/9/71 28. Fries BE, Simon SE, Morris JN, Flodstrom C, Bookstein FL: Pain in U.S. nursing homes: validating a pain scale for the minimum data set. Gerontologist 2001, 41(2):173-179. for long-term care in eight countries. Milbank Memorial Fund 2003:129. for long-term care in eight countries. Milbank Memorial Fund 2003:129. 7. Fries BE, Schneider DP, Foley WJ, Gavazzi M, Burke R, Cornelius E: Refining a case-mix measure for nursing homes: Resource 7. Fries BE, Schneider DP, Foley WJ, Gavazzi M, Burke R, Cornelius E: Refining a case-mix measure for nursing homes: Resource Utilization Groups (RUG-III). Med Care 1994, 32(7):668-685. 8 M V I O F BE Ph ll C T J H J H C M 7. Fries BE, Schneider DP, Foley WJ, Gavazzi M, Burke R, Cornelius E: R fi i i f i h R g ( ) 29. Hirdes JP, Frijters DH, Teare GF: The MDS-CHESS scale: a new measure to predict mortality in institutionalized older peo- ple. J Am Geriatr Soc 2003, 51(1):96-100. Refining a case-mix measure for nursing homes: Resource Utilization Groups (RUG-III). Med Care 1994, 32(7):668-685. p ( ) ( ) 8. Mor V, Intrator O, Fries BE, Phillips C, Teno J, Hiris J, Hawes C, Mor- ris J: Changes in hospitalization associated with introducing the Resident Assessment Instrument. J Am Geriatr Soc 1997, 45(8):1002-1010. p J ( ) 30. Bjorkgren MA, Fries BE, Arbor A, Shugarman LR: A RUG-III case- mix system for home care. Canadian Journal on Aging 2000, 19(Supplement 2):106-125. ( ) 9. Mor V: Improving the quality of long-term care with better information. Milbank Q 2005, 83(3):333-364. ( pp ) 31. Hirdes JP: Addressing the health needs of frail elderly people: Ontario's experience with an integrated health information system. Age Ageing 2006, 35(4):329-331. 10. Implementing the resident assessment instrument: Case studies of policymaking for long-term care in eight countries [http://www.milbank.org/reports/interRAI/030222interRAI.html] Authors' contributions p f g 4. Wagner EH, Austin BT, Von Korff M: Organizing care for patients with chronic illness. Milbank Q 1996, 74(4):511-544. LG prepared the initial manuscript, and integrated contri- butions of other authors. All authors made substantial written contributions to the manuscript, reviewed at least 2 revisions, and have given approval to the final version presented here. JM headed the research program that lead to the development of the instrument suite. Other authors Q ( ) 5. Hirdes JP, Fries BE, Morris JN, Steel K, Mor V, Frijters D, LaBine S, Schalm C, Stones MJ, Teare G, et al.: Integrated health informa- tion systems based on the RAI/MDS series of instruments. Healthc Manage Forum 1999, 12(4):30-40. g ( ) 6. Fries BE, Fahey CJ, Hawes C, Vladeck BC, Morris J, Phillips C, Fredek- ing H, Hirdes J, Sinclair DG, King JT, et al.: Implementing the Res- ident Assessment Instrument: Case studies of policymaking g ( ) 6. Fries BE, Fahey CJ, Hawes C, Vladeck BC, Morris J, Phillips C, Fredek- ing H, Hirdes J, Sinclair DG, King JT, et al.: Implementing the Res- ident Assessment Instrument: Case studies of policymaking Page 9 of 10 (page number not for citation purposes) BMC Health Services Research 2009, 9:71 http://www.biomedcentral.com/1472-6963/9/71 Pre-publication history Carpenter GI, Gill S, et al.: A Comparison of MDS/RAI Activity of Daily Living, Cognitive Performance and Depression Scales with widely used scales. Age and Ageing 1999, 28(S2):29. J y ( ) 26. Carpenter GI, Gill S, et al.: A Comparison of MDS/RAI Activity of Daily Living, Cognitive Performance and Depression Scales with widely used scales. Age and Ageing 1999, 28(S2):29. 27. Burrows AB, Morris JN, Simon SE, Hirdes JP, Phillips C: Develop- ment of a minimum data set-based depression rating scale for use in nursing homes. Age and ageing 2000, 29(2):165-172. y g g g ( ) 27. 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https://openalex.org/W1563937250	https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0126462&type=printable	English	null	Serotonin Depletion-Induced Maladaptive Aggression Requires the Presence of Androgens	PloS one	2,015	cc-by	7,358	Serotonin Depletion-Induced Maladaptive Aggression Requires the Presence of Androgens Erik Studer1, Jakob Näslund1, Erik Andersson1, Staffan Nilsson2, Lars Westberg1, Elias Eriksson1* 1 Department of Pharmacology, Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, POB 431, SE 405 30 Gothenburg, Sweden, 2 Mathematical Sciences, Chalmers University of Technology, SE 412 96 Gothenburg, Sweden * elias.eriksson@neuro.gu.se * elias.eriksson@neuro.gu.se OPEN ACCESS Citation: Studer E, Näslund J, Andersson E, Nilsson S, Westberg L, Eriksson E (2015) Serotonin Depletion-Induced Maladaptive Aggression Requires the Presence of Androgens. PLoS ONE 10(5): e0126462. doi:10.1371/journal.pone.0126462 Academic Editor: Cheryl McCormick, Brock University, CANADA Received: October 11, 2014 Accepted: April 2, 2015 Published: May 15, 2015 Copyright: © 2015 Studer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Academic Editor: Cheryl McCormick, Brock University, CANADA Academic Editor: Cheryl McCormick, Brock University, CANADA Published: May 15, 2015 Copyright: © 2015 Studer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All data are available from the Dryad digital repository (DOI: http://dx.doi. org/10.5061/dryad.b4d70). Funding: Financial support was obtained from the Swedish Science Council (EE, K2010-61x-4961—01- 3) http://www.vr.se/, Söderberg’s Foundation (EE, MT30/09) http://ragnarsoderbergsstiftelse.se/, Hållsten’s Foundation (EE), and the Brain Foundation (EE, FO2011-0293) http://www.hjarnfonden.se/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. RESEARCH ARTICLE Abstract The sex hormone testosterone and the neurotransmitter serotonin exert opposite effects on several aspects of behavior including territorial aggression. It is however not settled if tes- tosterone exerts its pro-aggressive effects by reducing serotonin transmission and/or if the anti-aggressive effect of serotonin requires the presence of the androgen. Using the resi- dent intruder test, we now show that administration of the serotonin synthesis inhibitor para- chlorophenylalanine (300 mg/kg x 3 days) increases the total time of attack as well as the percentage amount of social behavior spent on attack but not that spent on threat – i.e. that it induces a pattern of unrestricted, maladaptive aggression – in gonadectomized C57Bl/6 male mice receiving testosterone replacement; in contrast, it failed to reinstate aggression in those not given testosterone. Whereas these results suggest the pro-aggressive effect of testosterone to be independent of serotonin, and not caused by an inhibition of serotonergic activity, the pCPA-induced induction of maladaptive aggression appears to require the pres- ence of the hormone. In line with these findings, pCPA enhanced the total time of attack as well the relative time spent on attacks but not threats also in wild-type gonadally intact male C57Bl/6 mice, but failed to reinstate aggression in mice rendered hypo-aggressive by early knock-out of androgen receptors in the brain (ARNesDel mice). We conclude that androgenic deficiency does not dampen aggression by unleashing an anti-aggressive serotonergic in- fluence; instead serotonin seems to modulate aggressive behavior by exerting a parallel- coupled inhibitory role on androgen-driven aggression, which is irrelevant in the absence of the hormone, and the arresting of which leads to enhanced maladaptive aggression. * elias.eriksson@neuro.gu.se Serotonin Dampens Testosterone-Dependent Aggression paradigms) [2,3] and sexual behavior [4] in rodents, serotonin appears to exert the opposite ef- fects on all these aspects of behavior [5–7]. Likewise, whereas lack of testosterone often impairs aggressive behavior, serotonin is reported to reduce aggression, and particularly to counteract so-called maladaptive aggression [8]. Competing Interests: The authors have declared that no competing interests exist. This striking functional antagonism between testosterone and serotonin with respect to be- havior, in conjunction with the observation that testosterone exerts a profound influence on brain serotonergic activity [9–14], makes it tempting to suggest that the steroid might exert its behavioral effects by reducing the activity of serotonergic pathways. According to this view, the reduction in aggression and sexual activity observed in androgen-deficient animals would be the result of an anti-aggressive influence of serotonin being unleashed by the removal of the an- drogen. Alternatively, however, androgen-dependent behavior, including aggression, may be under the influence of a parallel, independent negative modulation by serotonin; to dampen or modulate sex steroid-driven behavior may hence constitute an important physiological role of this transmitter [15,16]. Finally, a third possibility would be that serotonin influences aspects of behavior that also happen to be under the influence of sex steroids, but would exert this in- fluence also in the absence of the hormone. Although many previous reports have discussed the influence of testosterone and serotonin, respectively, on the regulation of aggression, as well as the possible interplay between the two [13,17–23], there are, to our knowledge, no previous reports examining to what extent aggres- sion induced by testosterone at physiological doses in gonadectomized (GDX) animals is sero- tonin-independent, i.e. if testosterone promotes aggression also in the absence of serotonin, which would not be the case if testosterone stimulates aggression mainly by dampening a tonic inhibitory serotonergic influence. Reciprocally, it is also not known if presence of testosterone is an indispensable prerequisite for the well-established aggression-provoking effect of arrest- ing serotonergic transmission. The resident intruder (RI) paradigm is an animal model reflecting offensive, territorial ag- gression in which testosterone and serotonin have previously been shown to exert pro-and anti-aggressive effects, respectively [11,22]. Aggressive behavior exerted by the resident versus the intruder in this test might be viewed as a form of functional social communication and should, as such, follow certain patterns. Introduction The male sexual hormone testosterone and the brain neurotransmitter serotonin exert opposite effects on many aspects of behavior. Thus, whereas testosterone has been shown to promote territorial behavior (such as urine spraying) [1], impulsivity (as assessed using various conflict 1 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression To this end, mice were rendered non-aggressive by means of orchidectomy and then ex- posed to implantation of slow-release testosterone pellets or sham pellets, followed by the ad- ministration of either vehicle or the irreversible inhibitor of tryptophan hydroxylase, para- Chlorophenylalanine (pCPA), at a regimen previously shown to effectively deplete brain sero- tonin [24] as well as to increase aggression [25]. If the pro-aggressive effect of testosterone were mediated by a direct or indirect reduction of an anti-aggressive serotonergic input, one would expect serotonin depletion to reinstate aggression in gonadectomized (GDX) mice; moreover, administration of testosterone would fail to cause any further increase in aggression in pCPA- treated animals (when compared to pCPA-treated GDX mice). On the other hand, if serotonin reduces testosterone-dependent aggression by a parallel inhibitory path, serotonin depletion would not be sufficient to reinstate aggression after gonadectomy, and pCPA would enhance rather than mask the pro-aggressive effect of the hormone. Finally, if serotonin dampens tes- tosterone-independent aggression, pCPA would be expected to enhance aggression in GDX mice regardless of whether they are the subject of testosterone replacement or not. Also, by ex- amining if pCPA enhances actual attacks rather than threats, we were able to address the theo- ry that a primary role of serotonin in this context is to dampen maladaptive rather than functional aggression. Although the pro-aggressive effects of testosterone partly seems to be mediated by its me- tabolite estradiol acting via estrogen receptors (ER) [26], also AR activation is required for nor- mal aggressive behavior [27] as illustrated, e.g., by previous studies on male mice lacking functional ARs in the central nervous system (ARNesDel) [28,29]. In a second experiment, we wanted to shed further light on the possible interactions between serotonin and androgens for the regulation of aggression by exploring if the reduction in aggression following this form of reduced androgenicity is serotonin-dependent; to this end, it was examined if pCPA can rein- state aggressive behavior in ARNesDel mice or if serotonin depletion enhances aggression in wild-type controls only. Moreover, it was again assessed if aggression provoked by pCPA is of a maladaptive kind. To fulfill its purpose it should not comprise an exces- sive number of actuals attacks when threats of attacks would be sufficient for the aim to be met, i.e. for eliciting submissive postures from the intruder signaling maintained territorial control and dominance by the resident. Moreover, the behavior should not continue once this goal has been reached. In contrast, aggression characterized by a relative paucity of threat signals versus actual at- tacks and short latency to attack have recently been described as maladaptive [8]. While recent studies suggest that serotonin dampens such maladaptive (rather than functional) aggression, there are, to our knowledge, no studies exploring if the aggression induced by serotonin deple- tion in the resident intruder paradigm is largely of a maladaptive, dysfunctional kind, i.e. if ar- resting serotonergic activity elicits more attacks than threats. The first part of this study had two purposes. Firstly we wanted to address three different possibilities with respect to the interplay between serotonin and testosterone in the regulation of aggression: i) that testosterone promotes aggression by inhibiting serotonergic transmission, ii) that serotonin instead exerts a parallel inhibitory influence on testosterone-dependent ag- gression (of relevance only in the presence of the androgen) and iii) that serotonin dampens ag- gression regardless of whether testosterone in present or not. Secondly we wanted to address the hypothesis that aggression induced by removal of serotonin primarily is of a maladaptive kind, i.e. that serotonin depletion promotes attacks rather than threats. 2 / 12 Zfy: 5´- AAG ATA AGC TTA CAT AAT CAC ATG GA—3´ and 5´- CCT ATG AAA TCC TTT GCT GCA CAT GT—3´. In total eight ARNesDel mice were available for the experiment. Zfy: 5´- AAG ATA AGC TTA CAT AAT CAC ATG GA—3´ and 5´- CCT ATG AAA TCC TTT GCT GCA CAT GT—3´. In total eight ARNesDel mice were available for the experiment. As controls we used 11 littermate wild-type mice and eight AR-flox mice bearing only the LoxP-site but not expressing cre (ARloxP); both these groups should have intact AR activity, and they have previously been shown to display similar levels of aggression [29]. As expected, there were no differences between wild-type and ARloxP with respect to any of the behavioral parameters so these groups were collapsed and used as one AR-intact control group in the sta- tistical analyses. The animals were six to eight months of age when tested. They were moved to individual housing 24 days prior to the first tests of aggressive behavior. Housing of animals. In both experiments, animals were kept under controlled conditions at the Laboratory for experimental biomedicine at the University of Gothenburg. They were housed in standard vivarium cages (375 x 215 x 150 mm) with free access to food and water on a 12/12 light/dark cycle. 2.2 pCPA treatment In experiment I, gonadectomized mice given testosterone replacement as well as those without such replacement were administered either saline or pCPA methyl ester hydrochloride (300 mg/kg in saline solution buffered to pH 6) intraperitoneally once daily for three consecutive days. In experiment II, the same treatment was given to ARNesDel mice and wild type controls, respectively. 2.4 Ethics All procedures were subjected to approval by the Ethical Committee on Animal Experiments, Gothenburg, Sweden (permit numbers: 34–11, 67–12) and performed accordingly. All surgery was performed under general anesthesia and animals were allowed to recover in temperature- controlled cages, with additional monitoring for 3 days to ensure complete recovery. 2.3 Resident intruder test Animals were subjected to a standard RI test as previously described [29]. Briefly, the mice were tested during the dark phase in their home cage against an intruder mouse for 15 minutes. As intruders served male mice of the 129/SvEv strain (Taconic Farms, Denmark) that were kept group-housed throughout the experiment. The intruders were weighed before each behav- ioral test to ensure that they were at least 0.5 g lighter than the resident, with each intruder used once daily. The test was performed twice, first to obtain a baseline assessment before drug administration, and then again 24 h after the last injection of pCPA or saline, respectively. The tests were videotaped with an overhead video camera under IR illumination for subsequent behavioral analysis. 2.1 Animals Experiment I. Male C57Bl/6N mice (Charles River, Denmark), aged 10 weeks at arrival, were anaesthetized with a 3:12 vol/vol mixture of ketamine (Ketalar 10mg/ml, Pfizer) and xyla- zine (Rompun Vet 20 mg/ml, Bayer Animal Health), gonadectomized via a midline incision, and allowed to recover in groups of five. After a period of two weeks, aimed at allowing hor- monal levels to decline and the effect of orchidectomy on aggressive behavior to be fully devel- oped [30], the mice were anaesthetized as described above, and implanted with slow-release pellets containing 15 mg of testosterone (T, n = 18) or vehicle (GDX, n = 15) and designed for 60 days of even release (250 μg/day) (Innovative Research of America, USA). After this opera- tion, the animals were allowed to recover for two weeks before they were moved to individual cages. After nine days of isolation, the first RI test was undertaken. Experiment II. Generation of ARNesDel has been described in detail elsewhere [28]. In brief, male C57Bl/6 mice expressing Cre driven by the neuronal Nestin promoter were ob- tained from Jackson Laboratory (Bar Harbor, Maine, US) and mated with female mice carrying LoxP-sites [31] flanking the second exon of the androgen receptor gene which had been back- crossed into the C57Bl/6 background for at least 6 generations prior to the arrival at our labora- tory [32]. Genotypes were confirmed with PCR using the following primers: AR: 5´-AGC CTG TAT ACT CAG TTG GGG- 3´ and 5´-AAT GCA TCA CAT TAA GTT GAT ACC- 3´, Cre: 5´-GTTCGCAAGAACCTGATGGACA-3´and 5´-CTAGAGCCTGTTTTGCACGTTC-3´, 3 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression Zfy: 5´- AAG ATA AGC TTA CAT AAT CAC ATG GA—3´ and 5´- CCT ATG AAA TCC TTT GCT GCA CAT GT—3´. In total eight ARNesDel mice were available for the experiment. As controls we used 11 littermate wild-type mice and eight AR-flox mice bearing only the LoxP-site but not expressing cre (ARloxP); both these groups should have intact AR activity, and they have previously been shown to display similar levels of aggression [29]. As expected, there were no differences between wild-type and ARloxP with respect to any of the behavioral parameters so these groups were collapsed and used as one AR-intact control group in the sta- tistical analyses The animals were six to eight months of age when tested They were moved to 2.5 Data analysis Scoring was performed at low video speed by a trained observer blind to the status of the mice. Duration of all social exploration (including sniffing on the body or head of the intruder, ano- genital sniffing, social grooming and following), threat behaviors (including tail rattling, all of- fensive postures and aggressive grooming) and attack behaviors (including bite, chase and wrestling) were recorded. Mice not displaying at least one episode of attack or threat were char- acterized as non-aggressive. In addition, mice of different groups were compared with respect to total duration of attacks as well as latency to first attack; if no attack could be detected during the entire duration of the test, attack latency was set to 900 seconds. Finally, the behavioral 4 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression pattern of the animals was revealed by calculating the percentage of each category of behavior (i.e. social exploration, threat and attack, respectively) in relation to the total time of social in- teraction [33]. pattern of the animals was revealed by calculating the percentage of each category of behavior (i.e. social exploration, threat and attack, respectively) in relation to the total time of social in- teraction [33]. 2.6 Statistics Fisher’s exact test was used to compare testosterone treated GDX mice versus vehicle treated GDX mice (experiment I) and wild-type mice versus ARNesDel mice (experiment II) with re- spect to the number of mice displaying aggressive behavior (as defined in the section 2.5). In addition, comparison between the groups with respect to duration of attacks and latency to at- tack at the first test of each experiment was undertaken using unpaired t-test, or Welch's test for unequal variance when such variances between the groups were revealed by Levene’s test. tack at the first test of each experiment was undertaken using unpaired t-test, or Welch's test for unequal variance when such variances between the groups were revealed by Levene’s test. Since pCPA failed to evoke aggression both in GDX mice with no testosterone replacement and in ARNesDel mice, a further analysis of the possible effect of serotonin depletion on aggres- sion in the second test was performed only for testosterone-treated GDX mice of experiment I and wild-type mice of experiment II. Since pCPA failed to evoke aggression both in GDX mice with no testosterone replacement and in ARNesDel mice, a further analysis of the possible effect of serotonin depletion on aggres- sion in the second test was performed only for testosterone-treated GDX mice of experiment I and wild-type mice of experiment II. For both experiments, possible differences with respect to total time of attacks and latency to attack between animals treated with pCPA and saline, respectively, were analyzed using a general linear model for between-subject effects with the behavior in the first test as a covariate predictor. In addition, to explore to what extent pCPA induced maladaptive aggression as indi- cated by an increase in attacks rather than threats, the relative time spent on various forms of social behavior (social exploration, threat and attack, respectively) was calculated and the be- havioral pattern displayed at baseline compared to that displayed after treatment with pCPA or saline. In order to separate effects of repeated testing and treatment, respectively, a linear mixed model, with test and treatment as independent factors, was used for this purpose. Th h t th d t t d ±S E M Throughout the paper data are presented as group means±S.E.M. PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 3.2 Effect of pCPA on aggressive behavior in mice with reduced androgen levels or reduced androgen receptor expression None of the 7 GDX animals without testosterone replacement displayed aggression after treat- ment with pCPA (Table 1B). Likewise, pCPA failed to provoke aggression in ARNesDel mice (Table 2B). Serotonin Dampens Testosterone-Dependent Aggression Table 1. Aggressive behavior in Experiment I. A) Testosterone (n = 18) Vehicle (n = 15) No. aggressive mice (%) 16/18 (89) 1/15 (7)### Time attacking (s) 31.0 ±7.4 5.3 ±5.3## B) Saline (n = 9) pCPA (n = 9) Saline (n = 8) pCPA (n = 7) No. aggressive mice (%) 9/9 (100) 9/9 (100) 1/8 (12.5)* 0/7 (0)* A) Number of testosterone or vehicle treated GDX mice displaying aggression in the ﬁrst resident intruder test and the total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. # indicates level of signiﬁcance versus testosterone treated mice; ## p < 0.01, Table 1. Aggressive behavior in Experiment I. Table 1. Aggressive behavior in Experiment I. A) Number of testosterone or vehicle treated GDX mice displaying aggression in the ﬁrst resident intruder test and the total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. # indicates level of signiﬁcance versus testosterone treated mice; ## p < 0 01 A) Number of testosterone or vehicle treated GDX mice displaying aggression in the ﬁrst resident intruder test and the total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. # indicates level of signiﬁcance versus testosterone treated mice; ## A) Number of testosterone or vehicle treated GDX mice displaying aggression in the ﬁrst resident intruder test and the total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. # indicates level of signiﬁcance versus testosterone treated mice; ## p < 0.01, p ### p < 0.001. p 0.001. * indicates level of signiﬁcance versus the corresponding group of testosterone treated mice; *** p < 0.001. doi:10.1371/journal.pone.0126462.t001 3.1 Effect of treatments reducing androgen levels or androgen receptor expression on aggressive behavior As expected, at test 1 GDX mice not receiving testosterone replacement displayed aggression to a very limited degree, only 1 out of 15 being defined as aggressive. In contrast, in the group re- ceiving testosterone replacement, 16 out of 18 displayed aggressive behavior (Table 1A). This difference in number of mice being aggressive was reflected also by a difference with respect to duration of attack (GDX: 5.3±5.3 s, n = 15, GDX+T: 31.0±7.4 s, n = 18; t29.6 = -2.8; p<0.01), number of attacks (GDX: 1.5±1.5, n = 15; GDX+T: 12.8±3.1, n = 18; t23.9 = -3.3; p<0.01) and latency to attack (GDX: 876±23.8 s, n = 15; GDX+T: 369±70.8 s, n = 18; t20.7 = -6.8; p<0.001). While the number of mice displaying aggression at test 1 in experiment II did not differ be- tween groups (Table 2A), ARNesDel differed from controls both with respect to time attacking (ARNesDel: 7.2 ±4.0 s, n = 8; controls: 45.2±12.1 s; n = 20; t22.6 = −3.0; p<0.01) and with respect to attack latency (ARNesDel: 672±109 s; n = 8; t26 = 2.2; controls: 372±75.7 s; n = 20; p<0.05); with respect to total number of attacks, this difference however did not reach the level of statis- tical significance (controls: 8.4±2.3, n = 20; ARNesDel: 2.9±1.5, n = 8; t26 = -1.5; p>0.05). 5 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 3.3 Effects of pCPA on aggressive behavior in testosterone-treated GDX mice in experiment I and in wild-type mice in experiment II While pCPA failed to increase aggression in GDX mice not replaced with testosterone (experi- ment I) as well as in ARNesDel mice (experiment II), a general linear model revealed pCPA treatment to increase total time of attack in testosterone-replaced GDX mice in experiment I (F1,15 = 11.6; p<0.01; Fig 1A) and in wild-type mice in experiment II (F1,16 = 5.1; p<0.05; Fig 1B). However, while pCPA also shortened latency in experiment II (saline: 442.2±124.7 s; n = 10; pCPA: 78.7±17.9 s; n = 9; F1,16 = 11.8, p<0.01), the corresponding difference did not reach significance in experiment I (saline: 215.1 ±103.8 s; n = 9; pCPA: 64.4 ±21.8 s; n = 9; F1,15 = 1.9; p = 0.19). A mixed model analysis regarding the relative time spent on various forms of social behav- ior before or after treatment with pCPA or saline revealed pCPA to induce a pattern of Table 2. Aggressive behavior in Experiment II. A) Control (n = 20) ARNesDel (n = 8) No. aggressive mice (%) 15/20 (75) 4/8 (50) Time attacking (s) 45.2 ±12.1 7.2 ±4.0# B) Saline (n = 10) pCPA (n = 9)a Saline (n = 4) pCPA (n = 4) No. aggressive mice (%) 8/10 (80) 9/9 (100) 0/4 (0)* 0/4 (0)* A) Number of untreated control mice (WT and ARloxP) and ARNesDel mice displaying aggression in the ﬁrst resident intruder test and total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. Table 2. Aggressive behavior in Experiment II. Table 2. Aggressive behavior in Experiment II. Table 2. Aggressive behavior in Experiment II. Table 2. Aggressive behavior in Experiment II. A) Control (n = 20) ARNesDel (n = 8) No. aggressive mice (%) 15/20 (75) 4/8 (50) Time attacking (s) 45.2 ±12.1 7.2 ±4.0# B) Saline (n = 10) pCPA (n = 9)a Saline (n = 4) pCPA (n = 4) No. aggressive mice (%) 8/10 (80) 9/9 (100) 0/4 (0)* 0/4 (0)* A) Number of untreated control mice (WT and ARloxP) and ARNesDel mice displaying aggression in the ﬁrst resident intruder test and total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. doi:10.1371/journal.pone.0126462.t002 niﬁcance versus the corresponding group of controls; * p < 0.05 CPA treated group died prior to the second test of aggression. p e corresponding group of controls; * p < 0.05 signiﬁcance versus controls; # p < 0.05 3.3 Effects of pCPA on aggressive behavior in testosterone-treated GDX mice in experiment I and in wild-type mice in experiment II A) A) Number of untreated control mice (WT and ARloxP) and ARNesDel mice displaying aggression in the ﬁrst resident intruder test and total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA. A) Number of untreated control mice (WT and ARloxP) and ARNesDel mice displaying aggression in the ﬁrst resident intruder test and total duration of attack for each group. B) Number of mice displaying aggression in the second resident intruder test conducted 24 hours after the ﬁnal injection with saline or pCPA # indicates level of signiﬁcance versus controls; # p < 0.05 * indicates level of signiﬁcance versus the corresponding group of controls; * p < 0.05 * indicates level of signiﬁcance versus the corresponding group of controls; * p < 0.05 a One mouse in the pCPA treated group died prior to the second test of aggression. 6 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression maladaptive aggression in both testosterone-treated GDX mice (experiment I) and in wild typ mice (experiment II). In experiment I, the relative amounts of attacks but not threats were in- creased by serotonin depletion (% threat t(29 8) = 1 8 p>0 05; % attack t(31 9) = 3 0 p<0 01 Fig 1. Mean duration of attacks (±SEM) in mice given saline or pCPA (300 mg/kg/day for three days). The test lasted for 15 min. A. Testosterone-treated gonadectomized mice (saline: n = 9, pCPA: n = 9) (experiment I). B. Wild-type mice (saline: n = 10, pCPA: n = 9) (experiment II). * p < 0.05, ** p < 0.01 compared to mice treated with saline (general linear model). doi:10.1371/journal.pone.0126462.g001 Fig 1. Mean duration of attacks (±SEM) in mice given saline or pCPA (300 mg/kg/day for three days). The test lasted for 15 min. A. Testosterone-treated gonadectomized mice (saline: n = 9, pCPA: n = 9) (experiment I). B. Wild-type mice (saline: n = 10, pCPA: n = 9) (experiment II). * p < 0.05, ** p < 0.01 compared to mice treated with saline (general linear model). doi:10.1371/journal.pone.0126462.g001 maladaptive aggression in both testosterone-treated GDX mice (experiment I) and in wild type mice (experiment II). 3.3 Effects of pCPA on aggressive behavior in testosterone-treated GDX mice in experiment I and in wild-type mice in experiment II In experiment I, the relative amounts of attacks but not threats were in- creased by serotonin depletion (% threat, t(29.8) = 1.8, p>0.05; % attack, t(31.9) = 3.0, p<0.01; 7 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression % social, t(31.1) = −2.7, p<0.01; Fig 2A), and a similar effect was observed also in wild type mice in experiment II (% threat, t(32.8) = −0.4, p>0.05; % attack, t(22.0) = 3.9, p<0.001; % so- cial, t(21.8) = −2.4, p<0.05; Fig 2B). There was no effect of repeated testing in any of the experiments. % social, t(31.1) = −2.7, p<0.01; Fig 2A), and a similar effect was observed also in wild type mice in experiment II (% threat, t(32.8) = −0.4, p>0.05; % attack, t(22.0) = 3.9, p<0.001; % so- cial, t(21.8) = −2.4, p<0.05; Fig 2B). There was no effect of repeated testing in any of the experiments. % social, t(31.1) = −2.7, p<0.01; Fig 2A), and a similar effect was observed also in wild typ mice in experiment II (% threat, t(32.8) = −0.4, p>0.05; % attack, t(22.0) = 3.9, p<0.001; % cial, t(21.8) = −2.4, p<0.05; Fig 2B). There was no effect of repeated testing in any of the experiments. Fig 2. Comparison between the resident intruder baseline test (Test 1) and post-treatment test (Test 2) in animals receiving saline or pCPA (30 mg/kg/day for three days) with respect to mean percentage of the total time of social interaction spent on threat, attack and social behavior, respectively. The tests lasted for 15 min. A. Testosterone-treated gonadectomized mice (saline: n = 9, pCPA: n = 9) (experiment I). B. Wild-type mice (saline: n = 10, pCPA: n = 9) (experiment II). * p < 0.05, p < 0.01, * p < 0.001 against mice treated with (linear mixed model). doi:10.1371/journal.pone.0126462.g002 Fig 2. Comparison between the resident intruder baseline test (Test 1) and post-treatment test (Test 2) in animals receiving saline or pCPA (300 mg/kg/day for three days) with respect to mean percentage of the total time of social interaction spent on threat, attack and social behavior, respectively. The tests lasted for 15 min. A. Testosterone-treated gonadectomized mice (saline: n = 9, pCPA: n = 9) (experiment I). B. Wild-type mice (saline: n = 10, pCPA: n = 9) (experiment II). Discussion In line with previous studies, orchidectomized mice displayed very low levels of aggression [30] as compared to GDX animals exposed to testosterone replacement at a dose chosen to produce serum testosterone levels in the upper physiological range [34]. This difference being observed also in mice lacking serotonin by means of pCPA administration refutes the possibility that tes- tosterone promotes aggression by dampening an anti-aggressive serotonergic input. We hence conclude that serotonergic neurons, rather than being the mediator of the influence of testos- terone on aggression, exert a dampening influence on aggression that is parallel to the aggres- sion-promoting influence of the androgen. Further, the absence of pro-aggressive effect of pCPA in GDX animals without testosterone replacement indicates that the pro-aggressive ef- fect of pCPA [25] is relevant only in the presence of testosterone. Our results are compatible with previous studies showing long-term administration of a lower dose of pCPA to gonadally intact rats to enhance rather than mask aggression induced by the exposure to high doses of an- abolic androgens [13,35]. Where in the brain such a modulatory influence of serotonin on testosterone-dependent ag- gression takes place remains unknown. The medial amygdala, the hypothalamus, the prefrontal cortex and the lateral septum are however likely candidate areas, since these sites are all known to be involved in the regulation of aggression, and since they all display marked density of both sex steroid receptors and serotonergic nerve terminals [36–38]. While serotonin has often been attributed a general anti-aggressive influence, some authors have questioned this stance [8,39]. Instead, it has been suggested that a specific role for seroto- nin is to terminate aggression when the objectives of the aggressive behavior have been met [40], hence making the aggression functional and goal-directed. According to this theory, an impaired serotonergic transmission should primarily result in so-called maladaptive aggres- sion, i.e. by an increase in the number of attacks rather than threats [41]. While we could not address this possibility in GDX mice not receiving testosterone, since pCPA failed to reinstate aggression in these animals, we did indeed observe a pattern of disinhibited attack behavior, i.e. a relative increase in the percentage of time spent on attacks without a corresponding increase in time spent on threats, in testosterone-treated GDX mice. Notably, an almost identical effect of pCPA was observed also in the gonadally intact wild-type mice of experiment II. 3.3 Effects of pCPA on aggressive behavior in testosterone-treated GDX mice in experiment I and in wild-type mice in experiment II * p < 0.05, p < 0.01, * p < 0.001 against mice treated with (linear mixed model). Fig 2. Comparison between the resident intruder baseline test (Test 1) and post-treatment test (Test 2) in animals receiving saline or pCPA (300 mg/kg/day for three days) with respect to mean percentage of the total time of social interaction spent on threat, attack and social behavior, respectively. The tests lasted for 15 min. A. Testosterone-treated gonadectomized mice (saline: n = 9, pCPA: n = 9) (experiment I). B. Wild-type mice (saline: n = 10, pCPA: n = 9) (experiment II). * p < 0.05, p < 0.01, * p < 0.001 against mice treated with (linear mixed model). doi:10.1371/journal.pone.0126462.g002 8 / 12 PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Serotonin Dampens Testosterone-Dependent Aggression PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Acknowledgments The authors gratefully acknowledge the expert technical assistance of Ms. Inger Oscarsson and Ms. Gunilla Bourghardt. Professor Guido Verhoeven is gratefully acknowledged for providing us with ARloxP mice. Serotonin Dampens Testosterone-Dependent Aggression organizational effect of AR activation, or AR activation in the adult animal, that is necessary for pCPA to induce aggression, hence cannot be concluded from the present study. l A limitation of this study is that, for practical reasons, the number of ARNesDel mice admin- istered pCPA and saline, respectively, was low. However, given that PCPA did not elicit any ag- gression in any of the ARNesDel mice tested, it is highly unlikely that expanding the group would have led to a markedly different outcome. In conclusion, our data suggest that androgen-induced aggression is not masked by seroto- nin depletion, but, on the contrary, enhanced by pCPA, suggesting that testosterone does not exert its pro-aggressive effect by reducing serotonergic output. Instead the influence of seroto- nin on testosterone-induced aggression seems to be exerted by a parallel inhibitory pathway, the purpose of which seems to be to dampen maladaptive androgen-dependent aggression. Author Contributions Conceived and designed the experiments: ES EE LW. Performed the experiments: ES. Analyzed the data: ES EA SN. Contributed reagents/materials/analysis tools: EE LW. Wrote the paper: ES JN SN LW EE. ES JN SN LW EE. PLOS ONE \| DOI:10.1371/journal.pone.0126462 May 15, 2015 Discussion Of note is that treatment with pCPA also led to shortened attack latency, which may also be interpreted as a sign of maladaptive aggression [42] in experiment II, with a similar trend, though not sta- tistically significant, also in experiment I. It deserves to be mentioned that an increase in ag- gression was observed following the administration of the serotonin depleting agent in spite of the fact that both saline-treated GDX+testosterone mice and wild type animals displayed a rel- atively high amount of aggression as compared to what has been reported in some earlier stud- ies [e.g. 25]. The pro-aggressive effect of testosterone is to a high extent mediated by ERs but may require ARs as well. Supporting an involvement of the latter, we could confirm the result of previous studies [28,29] showing ARNesDel mice to display reduced aggression; since these animals display elevated testosterone and estrogen levels [28], and for this reason can be expected to display enhanced central ER activation, this observation clearly suggest that not only ER activa- tion, but also AR activation, is required for normal male aggression to be at hand. Likewise, our observation, though based on a small number of animals, that pCPA failed to enhance aggres- sion also in these mice, suggests that ER activation is not sufficient for serotonin depletion to provoke aggression, but that AR activation is required as well. 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https://openalex.org/W2511129736	https://jwcn-eurasipjournals.springeropen.com/counter/pdf/10.1186/s13638-016-0695-0	English	null	A cross-layer jitter-based TCP for wireless networks	EURASIP Journal on wireless communications and networking	2,016	cc-by	6,943	© 2016 The Author(s). Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. Keywords: Wireless TCP, Fairness, QoS, Performance evaluation Keywords: Wireless TCP, Fairness, QoS, Performance evaluation Keywords: Wireless TCP, Fairness, QoS, Performance evaluation A cross-layer jitter-based TCP for wireless networks Alessandro Andreadis* , Sandro Rizzuto and Riccardo Zambon Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 DOI 10.1186/s13638-016-0695-0 * Correspondence: andreadis@unisi.it Department of Information Engineering and Mathematics, University of Siena, Via Roma 56, 53100 Siena, Italy RESEARCH Open Access Open Access Abstract The Transmission Control Protocol (TCP) is one of the main communication protocols in the Internet, and it has been designed to provide an efficient reaction to packet loss events which are due to network congestion. Congestion is the main cause of losses in wired networks, but in today heterogeneous networks, loss events can also be introduced due to higher error rates on wireless channels, host mobility, and frequent handovers. Unfortunately, all packet losses are interpreted by TCP as a sign of congestion, triggering an inappropriate reaction which reduces its transmission rate and leads to performance degradation. In order to avoid this problem, it is important for TCP to correctly understand whether the reason of a packet loss is due to congestion or to a problem in the wireless link. This paper presents an innovative jitter-based cross-layer TCP algorithm, named XJTCP. It adopts the jitter ratio as loss predictor, joined with a layer two notification, in order to correctly infer the nature of a loss event. Performance evaluation and comparison with other common TCP implementations shows how XJTCP can be an interesting solution in the presence of wireless environments. 1 Introduction The different solutions proposed in literature can be categorized according to three broad approaches: split- connection, link-layer, and end-to-end approach [4, 5]. Split-connection protocols break TCP connection into two parts, at the base station; link-layer variants provide reliability through local retransmissions; finally, in end- to-end approaches, the sender performs loss recovery without breaking the semantics of TCP. The Transmission Control Protocol (TCP) is a reliable transport protocol widely adopted in today Internet. It adopts a suitable end-to-end congestion control scheme, providing high robustness against congestion losses, which constitute the main problem during packet trans- mission through wired networks [1]. In presence of het- erogeneous networks including wireless connections, packet losses can be caused not only by congestion events but also by random or burst errors on the wire- less channel. Nevertheless, TCP always reacts in the same way: it halves down its congestion window and triggers a backoff of its retransmission timeout (RTO). Such a reaction can lead to performance degradation when wireless links are present in the end-to-end path. This is the main reason why researchers have proposed several modified TCP protocols, at the aim to drive ap- propriate reactions when different types of losses are ex- perienced [2, 3] and, consequently, to improve TCP performance. Even if each methodology has several advantages and disadvantages, this work focuses on the end-to-end ap- proach because it does not involve any congestion expli- cit signaling from the network elements; therefore, it is highly deployable, with easy installation and utilization, and it requires modifications only in the terminal de- vices, without direct involvement of intermediate nodes. This paper introduces an innovative modification to the TCP protocol, named XJTCP, which is based on a cross-layer variant of the jitter-based TCP (JTCP) [6]. In particular, XJTCP adopts the inter-arrival jitter as loss predictor in order to infer whether a TCP packet loss is due to congestion or to random error. Moreover, it in- troduces a cross-layer mechanism for taking into ac- count the discarded medium access control (MAC) * Correspondence: andreadis@unisi.it Department of Information Engineering and Mathematics, University of Siena, Via Roma 56, 53100 Siena, Italy Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 2 of 11 frames, with the aim to enhance TCP performance over wireless networks. 2 TCP issues over wireless network TCP was originally designed for wired networks, where congestion is the main cause of losses and non-congestion loss events are usually negligible. Consequently, when a packet loss occurs, TCP effi- ciently reacts by reducing its congestion window size and retransmitting the lost data segment [1]. How- ever, the Internet of today is characterized by end-to- end paths which often include wireless links affected by higher error rates and variable delays [8]; the wire- less channel can generate non-congestion losses, due to either random bit errors or burst errors [2]. Given these issues, several TCP solutions have been proposed by researchers in order to improve TCP per- formance over heterogeneous networks [10]. At the time of writing, cross-layer approach can provide interesting solutions for TCP over wireless, since this approach does not violate the end-to-end semantics of TCP, by embed- ding all the modifications inside the sender and/or the receiver stack. On the other hand, it introduces more computational complexity, due to the interaction be- tween MAC and transport layers. Nowadays, one of the most diffused TCP implementa- tions is TCP Reno, which introduces fast recovery in conjunction with fast retransmit in order to avoid slow start phase after a single segment loss. Fast recovery is performed after receiving three duplicate acknowledg- ments (DACKs). Furthermore, the sender uses additional incoming DACKs to increase the congestion window, exiting the fast recovery phase when a unique ACK re- lated to new data is received. Then the congestion win- dow is set to half of the current congestion threshold and the DACK counter is set to zero. Reno’s fast recov- ery is optimized for the case when there is a single seg- ment loss within a congestion window [1]. Hence, it suffers from performance problems when multiple seg- ments are lost in the same congestion window. Random bit errors are often due to unpredictable ef- fects such as shadowing, fast fading, or interference, which can cause bit flipping in the transmitted data. When the MAC layer is not able to recover such errors, the received erroneous frame is discarded and TCP con- gestion control mechanism comes into play. Moreover, wireless channel condition is highly variable by nature, and this provokes delay fluctuations of the round-trip time (RTT), intended as the time between one packet is sent and its ACK is received. 1 Introduction In this environment, TCP inability to distinguish the actual reason (i.e., congestion or not) for a packet loss can lead to inappropriate reaction to a false congestion state, which results in wasteful behavior and unnecessary throughput decrease. Another key issue contributing to the degradation of TCP performance is packet reorder- ing, which is not such a rare event in wireless networks [3]. It happens when the receiving order of TCP packets differs from the sending order. High channel error rates in wireless links activate link-layer retransmissions which can lead to out-of-order delivery. The packet reordering problem violates the near in-order assumption made in the design of many traffic control mechanisms (i.e., du- plicate acknowledgments), introducing several unneces- sary retransmissions. The paper is organized as follows: Section 2 introduces the main problems affecting TCP protocol on wireless networks. Section 3 provides a detailed description of the proposed XJTCP algorithm and of its key features. Section 4 describes the simulation environment. Section 5 reports performance comparisons with popu- lar solutions such as Reno [1], JTCP [6], and Venoplus [7], under different loss typologies and congestion states. Finally, the last section concludes the paper with final remarks. 2 TCP issues over wireless network Since the TCP RTO is esti- mated on the basis of previous samples of the RTT, when such fluctuations exceed the estimated RTO, the in-flight packet is prematurely considered as it was lost, even if it is correctly received after the RTO deadline (i.e., spurious timeout), and an unnecessary retransmis- sion is triggered (i.e., spurious retransmission). Spurious retransmissions have two negative effects: first, they cause waste of bandwidth and secondly, they do not minimize energy consumption and that is an important issue due to limited power availability in wireless and mobile devices [9]. Jitter-based TCP (JTCP) adopts the jitter ratio “Jr” as a loss ratio predictor to determine the congestion level of the end-to-end path [6]. The jitter ratio is derived from the inter-arrival jitter between segments, as defined in real-time protocol (RTP). This information is deployed to investigate the link status, so as to take appropriate action on congestion window when a loss is detected. If the sender receives three DACKs or a RTO occurs, the value of Jr is compared with the inverse value of current congestion window size to distinguish between conges- tion and non-congestion losses. Then, if the loss is in- ferred as congestion loss, a Reno response is performed; otherwise, the congestion window is slightly reduced in order to maintain high throughput. Burst errors refer to a contiguous sequence of errone- ous bits, which are caused by persistent channel prob- lems, such as prolonged interference, long fading events, handovers and disconnections often due to user mobil- ity, and correlated packet losses are experienced. Conse- quently, TCP congestion control throttles its sending rate and reduces significantly its throughput. Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 3 of 11 Page 3 of 11 TCP Veno [11] joints the Reno mechanism with the channel estimation introduced in TCP Vegas, so as to distinguish between congestive and non-congestive losses. It adjusts the slow-start threshold according to the perceived network-congestion level. Specifically, it adopts a parameter β in order to infer congestion in the network: if the estimated congestion level exceeds the value of β, the loss is considered due to congestion and Reno additive-increase multiplicative-decrease (AIMD) scheme is applied. 3.1 Channel monitoring In order to monitor the wireless channel status and the nature of loss events, two variables are intro- duced, namely, Ci and Ni. Specifically, Ci is the con- gestion loss counter related to the i-th monitoring window, and it is incremented each time a loss occur- rence is inferred as associated to a congestion event. Ni is the non-congestion loss counter, and it is incre- mented each time a packet loss is not inferred as as- sociated to a congestion event (Fig. 1). Ni increase can be induced either by a cross-layer signaling within the receiver stack or by a transport layer event. In the first case (i.e., cross layer signaling), when a frame is discarded at the receiving MAC layer, such event is notified to the receiving TCP and, through the TCP option field, this information is piggybacked to the sender; consequently, the sender increases its Ni counter by one. The described end-to-end solutions try to distinguish congestion from random losses on the basis of some in- dications inferred at the transport layer, such as DACKs, timeouts and delay, or jitter, and they regulate their reac- tion by opportunely dimensioning the TCP congestion window. The algorithm here proposed still represents an end-to-end approach, thus not breaking the semantics of original TCP, but it also considers feedback which origi- nates from the MAC layer and is communicated to the transport layer. Due to this cross-layer signaling, the new TCP scheme can constantly monitor the channel status and it is aware of channel errors and related frame loss rates; consequently, it tries to gather the reason of packet loss, being it due to congested network or to channel impairments. The knowledge of the status of the wireless link at the moment in which the loss event is detected allows to better adjust the degree of TCP re- action and to better tune its congestion parameters. For this reason, the choice of a suitable algorithm capable of distinguishing whether network congestion or channel corruption is predominant in a wireless network appears of paramount importance, in order to achieve good per- formance in terms of throughput, delay, and jitter. 2 TCP issues over wireless network If the loss is assumed as due to ran- dom errors, Veno reduces the congestion window of 1/5. 3 XJTCP algorithm The key idea of the proposed algorithm is to integrate the jitter-based error estimation introduced in [6] with a cross-layer approach. Specifically, the XJTCP algorithm continuously monitors the wireless channel error status, by counting the number of inferred congestion and non- congestion loss events. According to this monitoring activity, the congestion control scheme takes an appropriate decision about TCP window size reduction and retransmissions. In particular, when three DACKs (TDACKs) or a RTO are detected, the algorithm evaluates whether it is a congestion or non-congestion loss and it adjusts the congestion win- dow opportunely. g TCP Venoplus [7] derives from Veno algorithm and it introduces two important refinements in order to distin- guish between congestion and random losses. Specific- ally, two new variables, congestion loss window, and random loss rate are adopted. Congestion loss window is incremented at each triple DACKs or timeout. Venoplus incorporates cross-layer functionality: it retrieves the re- ceived signal strength information (RSSI) and calculates BER for every packet received by the MAC layer, in order to evaluate the number of packet losses due to random errors in the wireless channel [7]. At each TCP segment loss, congestion loss window and random loss rate variables are used to opportunely reduce the con- gestion window (Cwnd). It infers segment loss due to congestion and random error (i.e., halving Cwnd value), due to random errors (i.e., Cwnd × 4/5), or due to con- gestion (i.e., halving Cwnd value). With respect to TCP Veno, TCP Venoplus improves the accuracy of conges- tion loss identification, providing significant enhance- ment on performance. 3.1 Channel monitoring In fact, IEEE 802.11 medium access control (MAC) layer [12] provides a checksum to prevent forwarding of erroneous frames, and a frame is discarded by the MAC layer according to one of the following reasons: the received signal strength indication (RSSI) is below a given threshold, frame check sequence (FCS) de- tects corruption, a frame (i.e., MAC) collision occurs. All such events denote that the loss event is due to channel problems and not to network congestion. Each transmitted MAC frame in wireless LANs should be promptly acknowledged, after a short inter- frame space (SIFS) time interval, if it has been cor- rectly delivered to the receiving station (only unicast frames are acknowledged). Nonetheless, random er- rors and collisions due to the channel contention mechanism can corrupt the transmission of the ori- ginal frame or of its returning ACK, and conse- quently, the ACK frame might not be received; so, the sender has to enter a new contention phase of the wireless channel for gaining the opportunity to Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 4 of 11 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 4 of 11 Fig. 1 Congestion loss and non-congestion loss counters retransmit the unacknowledged packet, until a max- imum number of attempts (i.e., retry limit parameter) is achieved. is very important because it determines how TCP should react to packet losses; as suggested in [6] for JTCP, the threshold is related to the value, in seg- ments, of the congestion window size (cwnd) when the loss occurs and can be given by Cross-layer signaling between MAC and TCP layers can be accomplished by interacting with the manage- ment information base (MIB) standard of IEEE 802.11 [12]. In particular, the indications about channel failures can be retrieved by looking inside the “dot11QosCoun- terTable” defined in this standard, as well as the number of lost packets in the downlink direction. As uplink is concerned, lost packets can be estimated from the num- ber of received MAC PDUs inside “dot11QosCounter- sTable” and the mean data rate field of the traffic specification (TSPEC) element. All such indications can be retrieved (e.g., through SNMP queries) at layer two and used at the TCP layer for increasing the non- congestion loss counter Ni. 3.1 Channel monitoring Th ¼ 1 cwnd = ð3Þ From the above considerations, if a loss event has been provoked by channel problems (i.e., non-congestion, Jr ≤ Th), Ni counter is increased. Otherwise, if Jr > Th, seg- ment loss is inferred as due to congestion and the Ci counter is incremented. Finally, in order to obtain a smoothed value of Ci and Ni, a low-pass filter is introduced at each monitoring window, by adopting the exponential weighted moving average (EWMA) [15], as shown in the following Eqs. (4) and (5): In the second case (i.e., transport layer event), when TDACKs or RTO occur, the TCP algorithm tries to infer whether such loss event has been caused by a congestion or by a channel problem. In detail, as in JTCP [6] and [13], the congestion state is deduced through a loss ratio predictor called jitter ratio (Jr), defined as: Cewmai ¼ αCi þ 1−α ð ÞCi−1 ð4Þ Newmai ¼ αNi þ 1−α ð ÞNi−1 ð5Þ ð4Þ ð5Þ where α is a smoothing factor, which denotes how recent and old samples of Ci and Ni influence the EWMA esti- mates. As an example, α = 0,8 means that 80 % of each new estimate comes from the last sampled value and only 20 % comes from the past sample (i.e., recent sam- ples are given more importance). Jr ¼ D i; j ð Þ Rj−Ri ¼ 1−Sj−Si Rj−Ri ð1Þ ð1Þ where Si, Ri and Sj, Rj are defined as the sending and re- ceiving times of segments i and j, respectively, and D(i,j) represents the inter-arrival jitter for the pair of packets (i, j) defined as the difference between the inter-packet times at the receiver and the inter-packet times at the sender [14]. 3.2 Regulation of the congestion window size A non-congestive loss can be detected at the MAC level or at the transport level. In the first case, the loss derives from a channel problem and this event is notified by the MAC layer through a cross-layer signaling to the receiv- ing TCP; such information can be used to increase Ni. D(i,j) can be computed as D i; j ð Þ ¼ Rj−Ri −Sj−Si ¼ Rj−Sj −Ri−Si ð Þ ð2Þ ð2Þ In the second case, the JTCP scheme infers whether the loss is due to congestion or not, and, consequently, the TCP sender decides which counter (i.e. Ci or Ni) should be increased. For each packet loss, XJTCP updates the inter- arrival jitter through Eq. (2) and calculates the Jr value through (1). If the jitter ratio is less than or equal to a given threshold Th, the segment loss is not considered congestion-driven; otherwise, it is due to congestion problems. Choosing the right threshold For each packet loss, XJTCP updates the inter- arrival jitter through Eq. (2) and calculates the Jr value through (1). If the jitter ratio is less than or equal to a given threshold Th, the segment loss is not considered congestion-driven; otherwise, it is due to congestion problems. Choosing the right threshold In order to better adjust the congestion window size and hence the aggressiveness of TCP reaction, the fol- lowing parameter θ has been introduced as follows: Page 5 of 11 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 θ ¼ Newmai þ 1 Cewmai þ 1 ð6Þ Fig. 2 Pseudo-code of XJTCP after TDACKs and RTO expiration ð6Þ θ represents the ratio between non-congestive loss rate and congestive loss rate. In (6), the unitary value is added in order to avoid the presence of poles in the function. The parameter θ helps to understand whether corrup- tion or congestion is the predominant condition in a certain time window. Consequently, the TCP congestion parameters (e.g. Cwnd) can be suitably adjusted, in order to regulate the aggressiveness of TCP scheme, thus in- creasing its throughput and reducing packet losses (e.g., if congestion is predominant, it might be better to trig- ger slow start phase rather than injecting further packets in a congested environment). 3.2 Regulation of the congestion window size So, θ > 1 indicates that non-congestive losses are pre- dominant in the wireless channel; thus, a loss event is expected to occur mainly due to a loss prone channel. On the other hand, if θ < 1, congestive losses are pre- dominant in the monitored window, and the TCP con- gestion control action is needed. Finally, if θ = 1, no clear indications can be obtained, and the reason of a loss event is inferred through the adoption of Jr, as for native JTCP algorithm. When three duplicate ACKs are received, if θ > 1, the event is considered caused by a lossy link and immediate recovery is performed, without any modification of con- gestion window and slow start threshold values. Immedi- ate recovery is also performed if θ = 1 and Jr ≤Th. Otherwise, if θ < 1, TDACKs are considered caused by a congestion, and fast recovery action is performed, by halving the congestion window and setting the slow start threshold to this new value. Fast recovery is also per- formed if θ = 1 and Jr > Th. After a RTO, if θ > 1 or θ = 1 and Jr ≤Th, fast recovery is performed, since loss is inferred due to non- congestive event. access point (AP); the AP communicates with a fixed host (i.e., the server) through two routers with wired links at 100 Mb/s. On the other hand, if θ < 1 or θ = 1 and Jr > Th, the oc- curred loss is inferred as a congestive event, thus trigger- ing a slow start phase. Figure 2 shows the pseudo-code of the described algorithm after three duplicate ACKs and retransmission timer expiration events. We assume that all STAs are positioned close to the AP (5–10 m distance), so as to constantly work at the maximum physical bit rate of 11 Mbit/s, and they are equipped with omni-directional antennas. The communication radius of the wireless area is about 250 m and the two-ray ground reflection model implemented in NS-2 is adopted. This radio propaga- tion model calculates the path loss when the received signal is composed by a direct line of sight path and a multi-path component which is mainly constituted by a single ground-reflected wave. It is shown [17] that this model gives more accurate prediction at a long distance than the free space model. 4 Simulation scenario To investigate XJTCP performance over wireless envi- ronments, several performance metrics (goodput, fairness, delay, jitter, and packet losses) have been ex- amined through extensive simulations which have been carried out through the popular NS-2 tool [16] (specifically, version 2.33). The simulation scenario, depicted in Fig. 3, consists of an IEEE 802.11b network in infrastructure mode, composed by five wireless stations (STAs) and an Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 6 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 6 of 11 Fig. 3 Simulation scenario Fig. 3 Simulation scenario traffic configurations have been tested, according to the desired level of congestion: All devices have the same buffer capacity of 1000 seg- ments, and the buffers are managed in a drop-tail policy. Each STA represents a single traffic source, and it can generate TCP or UDP packets, according to the follow- ing characteristics: No congestion: only one TCP traffic source (one STA) is active for the whole simulation time UDP congestion: one TCP traffic competes with four UDP streaming video connections, for the entire simulation time (five STAs are involved) The TCP traffic reproduces an uplink File Transfer Protocol (FTP), with a fixed packet size of 1024 byte (40 bytes header, 984 bytes payload) and an inter- packet generation time of 5 ms. Therefore, FTP can reach a maximum throughput of about 1.6 Mbit/s. TCP congestion: one TCP traffic under study (which can be Reno, JTCP, Venoplus, or XJTCP) competes with other four TCP Reno traffics (five STAs are involved), for the entire simulation. The choice of Reno version as a reference TCP is due to its popularity in almost all implementations The UDP traffic reproduces an uplink high quality video streaming, modeled as an on/off exponential traffic at variable bit rate (VBR). Traffic is generated according to an exponential distribution with an average rate of 1 Mb/s, burst time (“on” period) 1.2 s, idle time (“off” period) 1.8 s, and fixed packet size 512 bytes (8 bytes header, 504 bytes payload). During the on period, packets are generated at a constant rate of 2.5 Mbit/s, and during the off period, no packet is generated. 4 Simulation scenario The above configurations can be affected by different losses in the wireless link: Random losses, obtained by applying a uniform error model to the wireless channel Bursty losses (i.e., correlated losses), generated according to an exponential probability distribution: errors occur at time intervals which follow an exponential distribution function with a specific average error rate We have adopted TCP/FTP and UDP/VBR exponen- tial traffic models as implemented in NS-2 [16]. Three Fig. 4 Average goodput—random loss, no congestion Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 7 of 11 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 7 of 11 Fig. 5 Average goodput - random loss, UDP congestion Fig. 5 Average goodput - random loss, UDP congestion The packet error rate (PER) is different in all simula- tion runs (PER is set to 0, 1, 5, or 10 %). Results show that TCP versions based on jitter-ratio (XJTCP and JTCP) perform significantly better than Reno and Venoplus. This result can be justified mainly by the ability of distinguishing the cause of packet losses between congestion and corruption. In particular, with zero value of PER (ideal channel condition), the several versions of TCP exhibit equal performance, since the new mechanisms introduced by JCTP, XJTCP, and Veno- plus do not enter into play. This starting point is com- mon to all simulation configurations (i.e., no congestion and TCP/UDP congested network), and this is always true when PER = 0 %. The one-way time delay between the sender (wire- less station) and the receiver (fixed host) is set to about 50 ms, and the duration of the simulations is set to 300 s. Different TCP variants, namely TCP Reno, JTCP, VenoPlus, and XJTCP, have been compared under the described configuration sets. In order to study TCP be- havior under different traffic loads, the average TCP data transfer rate, named as average goodput, has been adopted as a reference parameter; its value has been ob- tained through repeated simulation runs (at least 10 in- dependent runs were made for each configuration). When PER grows, the performance difference is more evident, especially for the two jitter-based versions, since they are able to recognize that congestion is not present (due to a constantly low jitter) and thus their congestion window keeps higher than other TCPs. 4 Simulation scenario In fact, looking at the XJTCP pseudo-code of Fig. 2, no congestion con- dition always falls in the case θ > 1 and Jr ≤Th; therefore, XJTCP has the same reaction of JTCP (to random loss). 5 Performance evaluation The first series of simulations were performed in a wire- less channel affected by random losses; hence, a uniform error model has been implemented. In the first traffic configuration (no congestion), only one TCP traffic is active during the whole simulation. Figure 4 reports average goodput values obtained under different PER levels for each different TCP algorithm. When congestion is introduced (UDP congestion) through the activation of four UDP flows (Fig. 5), XJTCP achieves the maximum goodput under all PER levels, Fig. 6 Average goodput—random loss, TCP congestion Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 8 of 11 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 8 of 11 Fig. 7 Average goodput—bursty loss, no congestion Fig. 7 Average goodput—bursty loss, no congestion and this is due to its capacity of reacting differently in case of congestion and random losses. The difference between JTCP and XJTCP is now due to the introduc- tion of the θ parameter, which is now conditioned by both random losses and congested environment, thus ranging lower and above the unitary value. Conse- quently, XJTCP reaction can be different from JTCP and this affects its goodput positively. behave better in all configurations, but XJTCP, thanks to the θ indicator, achieves the best performance. In fact, in case of consecutive losses due to a burst channel, other TCP schemes react mainly to conges- tion (due to the fact that they can rarely receive TDACKs and their RTO expires), while XJTCP can still recognize a corrupted network and can enter into fast recovery mode (see Fig. 2, RTO expiration and θ > 1). Under TCP congestion (Fig. 6), the above consider- ations are still valid and XJTCP performs slightly better (about 5 %) than JTCP. Concerning the TCP congested scenario, XJTCP achieves again the best performance (Fig. 9), with a goodput increase of about 10 %, as before. When burst losses are introduced in the wireless chan- nel, under no critical conditions (no congestion) XJTCP (Fig. 7) performs similarly to the case of random losses, and jitter-based TCP versions perform better than Reno and Venoplus. Even if the described simulations provided good re- sults in terms of goodput, it is fundamental for new ver- sions of TCP algorithm not to behave unfairly when sharing connections with other TCP implementations. 5 Performance evaluation Therefore, fairness investigation related to XJTCP has been carried out in this work, by adopting the Jain’s fair- ness Index [18] as a benchmark. In the second configuration (UDP congestion), when the ability to distinguish between non-congestive and congestive losses becomes more important, XJTCP shows a substantial improvement with respect to its closest competitor (JTCP). In particular, its goodput performance (Fig. 8) performs better with non-zero PER values, up to 10 % more than JTCP. Even in this case, similarly to previous figures, jitter-based versions For this test, a TCP congested scenario has been adopted, where a XJTCP-based traffic competes with four TCP Reno traffics. Figure 10 shows the Jain’s fair- ness index trend for the XJTCP connection, under uni- form and bursty losses at different PER levels. Fig. 8 Average goodput—bursty loss, UDP congestion Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 9 of 11 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 9 of 11 Fig. 9 Average goodput—bursty loss, TCP congestion Fig. 9 Average goodput—bursty loss, TCP congestion Results show that XJTCP can coexist with traditional TCP implementations, showing a fair behavior that is more evident under random loss events. can favor XJTCP, which can better distinguish the reason of loss event. However, this phenomenon is still acceptable, and fairness keeps always above 94 % even under high PER values. However, in some cases, XJTCP can take advantage of the conservative approach of traditional TCP versions like Reno in case of non-congestive events. In fact, TCP Reno decision about the congestion level of the network is based only on TDACKs or timeout events. So, when non-congestive events increase (i.e., higher channel error rates), Reno algorithm can lead to an excessive reaction which causes repeated reductions of the Cwnd, with consequent release of network resources due to multiple slow start phases. The concurrent XJTCP can take ad- vantage of this fact, since it is designed to better inter- pret loss events, consequently it tends to exploit the released resources. Finally, further simulations have been carried out in order to compare the performance of the two jitter- based TCP algorithms (i.e., JTCP and XJTCP); lost packet, jitter, and delay metrics have been taken into ac- count for this comparison and results are shown in the Figs. 11, 12 and 13. 5 Performance evaluation The scenario of interest reproduces the worst network conditions, characterized by a bursty channel and UDP congestion configuration. Figure 11 shows the results related to lost packets for JTCP and XJTCP schemes. PER is considered at the link layer of 802.11. According to this standard, several cor- rupted frames can be recovered by link layer retransmis- sions; hence, TCP layer sees a more reliable channel, with a lower error rate. This is the reason why XJTCP fairness decreases at higher PER. Moreover, this phenomenon is more evi- dent in case of burst errors, when consecutive packet losses hinder receiving TDACKs and cause RTO ex- piration. Hence, this condition is misinterpreted by TCP Reno as a congestion problem, with unnecessary consecutive reductions of the Cwnd value and to drastic reduction of its packet injection rate. This fact JTCP exhibits a worse behavior (with a higher number of lost packets) than XJTCP. In fact, when PER aug- ments, JTCP is less capable than XJTCP to distinguish between congestion and corruption problems which caused the loss event. Therefore, TCP connections undergo many consecutive timeouts under a high jitter Fig. 10 Fairness Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 10 of 11 Page 10 of 11 Fig. 11 Lost packets—bursty loss, UDP congestion Fig. 11 Lost packets—bursty loss, UDP congestion The average delay increases when the number of re- transmitted packets grows and this happens more evi- dently in case of consecutive timeouts followed by slow start phases, which is the case of JTCP. Differently, XJTCP can take a more accurate decision about the pre- dominant condition in the network, and hence, it avoids to enter unnecessary slow start phases, with positive ef- fects on its average delay. value (due to congestion). This fact is interpreted by JTCP as pure congestion, and hence, it causes repetitive falls to the slow start phase. For this reason, the amount of packets to be retransmitted increases. On the contrary, XJTCP, thanks to the θ parameter, can better distinguish between congestion and corrup- tion and can react more suitably with fast recovery, thus experimenting an inferior number of slow start phases. Delay and jitter values can fluctuate due to reactions carried out at level 2 (i.e., retransmission by 802.11 MAC) or level 4 (i.e., TCP retransmissions and conges- tion control). 5 Performance evaluation If a packet is retransmitted more times, its delay increases, and consequently the jitter fluctuates. This fact impacts also on the average jitter depicted in Fig. 12, since the higher is the number of slow start phases, the more are the oscillations on the degree of congestion in the network and consequently the jitter values. Moreover, higher jitter values together with RTO expiration are interpreted by JTCP as a sign of pure con- gestion, triggering a reaction to slow start phase; this does not happen for XJTCP. The authors declare that they have no competing interests. The authors declare that they have no competing interests. Received: 1 January 2016 Accepted: 11 August 2016 6 Conclusions CP Fu, SC Liew, TCP Veno, TCP enhancement for transmission over wireless access networks. IEEE Journal of Selected Areas in Communications (JSAC) 21(2), 216–228 (2003) 12. IEEE Std. 802.11e-2005, Part 11: Wireless LAN Medium Access Control (MAC) and Physical Layer (PHY) specifications. Amendment 8: Medium Access Control (MAC) Quality of Service Enhancements, IEEE Std. 802.11e (2005) Further studies can investigate XJTCP behavior under different network technologies such as 3GPP-LTE and with user mobility. 13. A Andreadis, S Rizzuto, R Zambon, Performance comparison of JTCP over QoS- aware WiMAX networks, IEEE Symposium on Computers and Communications (ISCC), Corfu, Greece, pp.371-376 (2011) 13. A Andreadis, S Rizzuto, R Zambon, Performance comparison of JTCP over QoS- aware WiMAX networks, IEEE Symposium on Computers and Communications (ISCC), Corfu, Greece, pp.371-376 (2011) 14. H Schulzrinne, S Casner, R Frederick, V Jacobson, RTP: a transport protocol for real-time application, RFC 1889, Internet Engineering Task Force (IETF) (1996) 14. H Schulzrinne, S Casner, R Frederick, V Jacobson, RTP: a transport protocol for real-time application, RFC 1889, Internet Engineering Task Force (IETF) (1996) Acknowledgements Th h i h h The authors wish to thank the anonymous reviewers for their detailed and constructive comments, which helped so much to improve the quality of the paper. 15. W Richard Stevens, TCP/IP illustrated, Addison Wesley (1994) 16. Network Simulator ns-2. http://www.isi.edu/nsnam/ns. Accessed 17. T S Rappaport, Wireless communications, principles and practice. Prentice Hall (1996) References 1. M Allman, VE Paxson, W Stevens, TCP congestion control request for comments, RFC 2581, Network Working Group, Internet Engineering Task Force (1999) Submit your manuscript to a journal and beneﬁ t from: 7 Convenient online submission 7 Rigorous peer review 7 Immediate publication on acceptance 7 Open access: articles freely available online 7 High visibility within the ﬁ eld 7 Retaining the copyright to your article Submit your next manuscript at 7 springeropen.com Submit your manuscript to a journal and beneﬁ t from: 7 Convenient online submission 7 Rigorous peer review 7 Immediate publication on acceptance 7 Open access: articles freely available online 7 High visibility within the ﬁ eld 7 Retaining the copyright to your article Submit your next manuscript at 7 springeropen.com Submit your manuscript to a journal and beneﬁ t from: 6 Conclusions This work introduces a new TCP variant named XJTCP based on a cross-layer approach, and it inves- tigates its performance in presence of wireless net- works, compared with consolidated TCP solutions such as TCP Reno, JTCP, and VenoPlus. Several tests have been carried out, focusing on goodput and fair- ness parameters under random and burst losses and According to the above considerations, also average delay performance is affected. The average delay, shown in Fig. 13, has been calculated as the sum of the delays needed for each single packet to be correctly received, divided by the total number of received packets. Fig. 12 Average jitter—bursty loss, UDP congestion Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Andreadis et al. EURASIP Journal on Wireless Communications and Networking (2016) 2016:191 Page 11 of 11 Page 11 of 11 Fig. 13 Average delay—bursty loss, UDP congestion Fig. 13 Average delay—bursty loss, UDP congestion different congestion configurations. Performance com- parison has also been carried out between JTCP and XJTCP in terms of delay, jitter, and packet losses. Re- sults show that XJTCP exhibits a good performance in all configuration sets, thus representing an interest- ing solution when wireless scenarios are present in the end-to-end path. Even if the adopted cross-layer approach introduces new complexity in TCP algo- rithm, the computational power which characterizes today devices, included the mobile world, makes the implementation of XJTCP feasible without undermin- ing resources significantly. 7. S Shetty, Y Tang, W Collani, TCP venoplus—a cross-layer approach to improve TCP performance in wired-cum-wireless networks using signal strength, IEEE International Conference on Networking, Sensing and Control (ICNSC), Chicago (US), pp.693-697 (2010) 7. S Shetty, Y Tang, W Collani, TCP venoplus—a cross-layer approach to improve TCP performance in wired-cum-wireless networks using signal strength, IEEE International Conference on Networking, Sensing and Control (ICNSC), Chicago (US), pp.693-697 (2010) 8. 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IEEE Std. 802.11e-2005, Part 11: Wireless LAN Medium Access Control (MAC) and Physical Layer (PHY) specifications. Amendment 8: Medium Access Control (MAC) Quality of Service Enhancements, IEEE Std. 802.11e (2005) 10. A Afanasyev, N Tilley, P Reiher, L Kleinrock, Host-to-host congestion control for TCP. IEEE Commun. Surv. Tutorials. 12(3), 304–342 (2010) Submit your manuscript to a journal and beneﬁ t from: 2. KC Leoung, VOK Li, Transmission control protocol (TCP) in wireless networks: issues, approaches, and challenges. IEEE Commun. Surv. Tutorials. 8(4), 64–79 (2006) 3. B Sardar, D Saha, A survey of TCP enhancements for last-hop wireless t k IEEE C S T t i l 8(3) 20 34 (2006) 4. A Mammadov, B Abbasov, A Review of protocols related to enhancement of TCP performance in wireless and WLAN networks, IEEE 8th International Conference on Application of Information and Communication Technologies (AICT), pp.1-4, 15–17 (2014) 5. 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https://openalex.org/W2043810718	https://europepmc.org/articles/pmc2074056?pdf=render	English	null	Intracellular Localization of Carcinogen and its Relationship to the Mechanism of Carcinogenesis in Rat Liver	British journal of cancer	1,959	cc-by	8,125	236 236 INTRACELLULAR LOCALIZATION OF CARCINOGEN AND ITS RELATIONSHIP TO THE MECHANISM OF CARCINOGENESIS IN RAT LIVER SILVIO FIALA AND ANNA E. FIALA From the Department of Pathology, Columbia University and Francis Delafield Hospital, New York City Received for publication April 13, 1959 Received for publication April 13, 1959 THE experimental study of carcinogenesis aims at the finding of the common denominator for metabolic and structural changes which are characteristic of malignant transformation. In the previous paper (Fiala and Fiala, 1959) we tried to throw some light on this problem by correlating in time various changes during the carcinogenic process induced in rat liver by 3'-methyl, 4-dimethyl- aminoazobenzene. It was found that the whole process consists of two distinct phases. In the first there occurs a depletion of preformed cytoplasmic granules, accompanied by a lowering in the amount of respiratory activity, while glycolysis remains unchanged. Then, some 80 days after initiation of carcinogen feeding, a relatively sudden increase of anaerobic glycolysis takes place, coinciding with or slightly preceding the onset of massive cellular proliferation. At this time there occurs also a marked shift in the distribution of cytoplasmic ribonucleic acid from the ergastoplasm into the supernatant. g p p It is obvious, therefore, that the single factor we sought for is actually the ratio N +S K* M + CH + R- N +S K* M + CH + R- which changes its value in favor of the numerator in that critical period of carci- nogenesis which marks the onset of malignant transformation. If we substitute the active masses of subcellular fractions for the respective symbols, the result describes the great preponderance of the nuclei and unorganized soluble phase of the cell over the preformed cytoplasmic elements. This amounts to the lack of cytoplasmic differentiation. If, on the other hand we substitute the corres- ponding ribonucleic acid for each fraction, we obtain a shift in the distribution of ribonucleic acid from cytoplasmic elements into the nucleus and supernatant. When, finally, we substitute into this ratio the metabolic activities of the corres- ponding fractions, we obtain a sudden increase in glycolysis (concentrated in the supernatant) over and above the respiration of mitochondria. The positive Warburg quotient g q Q02 Co2 > 0 Qo2 Q02 Co2 > 0 Qo2 expressing the appearance of the aerobic glycolysis at this period is only an extreme case of this situation. * N = nucleus. M = mitochondria. CH = "fluffy layer ". R ergastoplasm. S soluble fraction. * N = nucleus. * N = nucleus. M = mitochondria. M = mitochondria. "fl ff l CH = "fluffy layer ". CH = "fluffy layer ". R ergastoplasm. y y R ergastoplasm. MATERIALS AND METHODS Sprague-Dawley rats and, in a few experiments, Swiss albino mice were used as experimental animals. Separation and purification of subcellular fractions was performed as described in the previous paper. p p p p Two criteria were used for estimating the purity of mitochondrial fractions: (a) lack of staining with fresh aqueous solution of Pyronin Y (0.1 per cent); (b) the basophilic quotient (B.Q.). The mitochondria apparently do not contain any ribonucleic acid (RNA) at all (Novikoff, 1956) or only small and variable amounts (Harel, Jacob and Moule, 1957). Their B.Q. (RNA/protein nitrogen) should be, therefore, close to zero. On the other hand the B.Q. of the ergastoplasmic fraction approaches 1. For this reason the B.Q. of the mitochondrial fraction indicates approximately the percentage of contamination with basophilic RNA-rich elements. Thus a B.Q. of 0.5 indicates that the mitochondria are approximately 50 per cent pure, while a B.Q. of 0.2 indicates 80 per cent. purity. per p , B.Q. 0. per p y The amount of protein-bound 3'-methyl, 4-dimethylaminoazobenzene (MDAB) was determined spectrophotometrically. The absorption of crystalline MDAB at pH 2.9 (absorption maximum at 520 m,.) was used as a standard. The protein fraction was hydrolyzed, extracted with ether, the ethereal extracts evaporated to near dryness and residuum redissolved in acid ethanol (Miller and Miller, 1947). 0 5 y ( e , 1947). Benzpyrene in 0.5 per cent benzene solution was painted twice on the hairless skin of 1-2 days old mice. The mice were sacrificed 24-48 hours later. The carci- nogen was visible by characteristic fluorescence in long UV-light. The mice 1-2 days old were chosen because the skin of older mice is too difficult to homo- genize. It is virtually impossible to separate their subcellular fractions because the broken tonofibrils adsorb indiscriminately all cytoplasmic elements. Even in young mice all cytoplasmic elements were heavily contaminated. The advantage of using very young mice for the study of carcinogen localization was confirmed recently by Calcutt (1958). The estimation of contamination of the mitochondrial fraction with ergastoplasm was based on the B.Q. The presence of the carci- nogen was determined by measuring the fluorescence of the hydrolyzed protein residue of each fraction, previously precipitated with 5 per cent trichloroacetic acid (TCA) and extracted in a Soxhlet apparatus with boiling ethanol for 48 hours (Miller, 1951). The results were compared with unpainted controls of norma mice of the same litter. MECHANISM OF CARCINOGENESIS IN LIVER If the process of carcinogenesis is conceived to be a shift from one equilibrium into another, then one or more factors must exist which keep this equilibrium within normal values. It seems plausible to assume that the protein-bound carcinogen interferes somehow with this regulating activity of the cell. In order to elucidate this fundamental problem of carcinogenesis, it is necessary to estab- lish the localization of the carcinogen within the cell. It seems obvious that any subcellular organelle with a predilection for the carcinogen is most likely its primary target and plays an important role in this regulating activity of the cell. The questions arising next are, therefore, what kind of processes are influenced by the localization of the carcinogen and how is the interference by the carcinogen with the regulating activity of the cell, leading to a new equilibrium at the critical period of carcinogenesis, concerned with the mechanism of this process. These problems outline the program of this paper. Received for publication April 13, 1959 g p S soluble fraction. 237 MECHANISM OF CARCINOGENESIS IN LIVER MATERIALS AND METHODS Fluorescence was measured using the fluorescent attachl- 238 SILVIO FIALA AND ANNA E. FIALA ment of a Beckman DU spectrophotometer and taking the fluorescence of 5 mg. of quinine sulfate in 25 ml. water as standard equal to 100 per cent of the total deflection. ment of a Beckman DU spectrophotometer and taking the fluorescence of 5 mg. of quinine sulfate in 25 ml. water as standard equal to 100 per cent of the total deflection. For the study of tryptophan peroxidase (TPO) L-tryptophan (78 mg. in 10 ml.) was injected intraperitoneally into halfofthe animals, while the other halfremained uninjected. Six hours after injection all animals were sacrificed. TPO was assayed spectrophotometrically (Knox, 1955) from metaphosphoric acid (5 per cent) extracts of liver samples incubated at 37° for 1 hour and from unincubated controls. The sediments obtained after precipitation with 15 per cent metaphosphoric acid were extracted following Schneider's (1945) procedure and the deoxyribonu- cleic acid (DNA) was measured by the diphenylamine reaction. In the experiments concerning the effect of the carcinogen on TPO formation, single massive doses of carcinogen (40 mg. of crystalline MDAB dissolved in 10 ml. of corn oil) were injected intraperitoneally into rats kept on a basal diet for four weeks prior to the experiment and a similar number of control animals were injected with corn oil free of carcinogen. Seventeen hours after intraperitoneal injection one half of the control animals and one half of the carcinogen injected animals were injected with tryptophan. yp p Glucose 6-phosphatase (G 6-Pase) activity was determined by measuring the amount of liberated inorganic phosphate by King's modification of the Fiske- Subba-Row procedure (King, 1932). Homogenates or separated ergastoplasmic fractions were diluted appropriately with Tris-buffer of pH 7.0 and G 6-P (1 ml. of approx. 1 X 10-3 M sol.) was added when incubation was started (37'). After 15 minutes the reaction was stopped by adding an equal volume of 10 per cent TCA. In unincubated controls TCA was added before G 6-P. The total volume of mixture was 3.0 ml. Polarographic determination of protein-bound sulphydryl groups was per- formed using Brdicka ammoniacal solution of cobaltous chloride as indicated in the previous paper. MATERIALS AND METHODS The proteins of the ergastoplasm were separated into soluble and insoluble fraction by the following procedure: the fraction spun down by 1 hour centrifuga- tion at 59000 x g (Spinco preparative ultracentrifuge) from the homogenate in isotonic sucrose (10 volumes) was resuspended in cold physiological saline and recentrifuged in a Servall SS-1 centrifuge for 2 hours at 30000 x g in the cold room. The supernatant was discarded, the sediment was resuspended in distilled water, dialyzed against distilled water for 48 hours at 4° and lyophilized. The lyophilized material was homogenized at pH 9.2 either in a cold Edsall (1930) solution (6 X 10-1KC1, 1 x 10-2M Na2C03, 4 x 10-2M NaHCO3) or, later, when it was seen that the reproducibility of the results demanded buffered solution, in Tris- buffer pH 9.1. The fraction was left for 1-2 hours in the cold with occasional stirring and then centrifuged for 1 hour at maximum speed in the Servall centrifuge. Both sediment, resuspended in distilled water, and supernatant were dialyzed against large volume of distilled water for 48 hours at 4°. The protein nitrogen of dialyzed subfractions was determined by Nesslerization. 1. Localization of Carcinogen Measurements of isolated subcellular fractions from livers of rats fed with MDAB have shown that purified nuclei contained practically no carcinogen. 1. Localization of Carcinogen RESULTS g Measurements of isolated subcellular fractions from livers of rats fed with MDAB have shown that purified nuclei contained practically no carcinogen. 239 MECHANISM OF CARCINOGENESIS IN LIVER The nuclear fraction did not stain pink after precipitation with trichloracetic acid throughout the period of carcinogenesis if basophilic cytoplasmic contaminates were removed by repeated washings with isotonic sucrose. Also the hydrolysis of the purified nuclear fraction did not liberate any carcinogenic azo dye. This demonstrated that no appreciable amount of carcinogen was present in the nuclear fraction. The localization and action of this carcinogen must, therefore, be cytoplasmic. y p The presence of the carcinogenic azo dye was noted in all cytoplasmic fractions after precipitation with 5 per cent TCA. The comparisons of amounts of MDAB liberated after hydrolysis of cytoplasmic fractions is shown in Table I. By far the greatest amount of this protein-bound carcinogen appeared in the soluble phase; considerably less was in the ergastoplasm; and very little was contained within the mitochondria. Still more significant was the fact that the purification of mitochondria, manifested in lowering of the B.Q. from 0.4 to 0.12 coincident with the removal of contaminating RNA-rich elements, resulted in a proportional loss of protein-bound carcinogen. The carcinogen content of liver mitochondria thus depended on contamination by ergastoplasm. TABLE I.-Distribution of MDAB in Cytoplasmic Fractions TABLE I.-Distribution of MDAB in Cytoplasmic Fractions Mitochondria A. II Fraction Supernatant Microsomes I (Purified) Protein nitrogen (mg.) 19.0 . 14 13 3 RNA (mg.) 11-4 15-4 . 5-2 0-36 B.Q.... 0-6 . 1.1 . 0.4 0-12 Dye liberated (g.) . 700 . 46-0 . 16-0 1.1 Specific dye binding . 3-62 . 3-3 1-2 0-36 Fig. 1 shows the absorption spectra of liberated azo dye from separated cyto- plasmic fractions. The spectrum calculated for mitochondria with B.Q. approaching zero disappears and is, therefore, drawn as a straight line. Under the idealized conditions of pure mitochondria, no carcinogen would be found. g The slight content of the carcinogen in the mitochondrial fraction dependent on the contamination with ergastoplasm, made clear why this carcinogenic azo dye is not a respiratory inhibitor. This result warranted investigation of the localization of a carcinogenic fluorescent hydrocarbon such as benzopyrene and methylcholanthrene because it was speculated for a long time that mitochondria are the main storage place of these carcinogens. Thus Graffi (1939) treated various cells in vitro with benzopyrene and methylcholanthrene dissolved in glycerol and serum and observed by fluorescence their accumulation inside mitochondria stained by Janus Green. The obvious drawbacks of his observation were that he used in vitro experiments, which are often less conclusive than those in vivo, that he chose for his study cells in which benzopyrene and methylcholanthrene do not produce tumors. In our experiments we have found that the location of benzopyrene in mouse skin depends on the mode of application. When the carcinogen was applied in lanoline, all cytoplasmic fractions yielded characteristic fluorescence in their alcoholic extracts while no carcinogen seemed to be liberated from protein residua after hydrolysis. On the other hand when the carcinogen was applied in benzene 240 SlLVIO FIALA AND ANNA E. FIALA solution, a significant fluorescence could be observed, even after a thorough alcohol extraction, from hydrolyzed protein residua ofthe supernatant and ergasto- plasmic fractions. In the homogenate obtained from 2g. of mouse epidermis, the supernatant was the most strongly fluorescent; the nuclear fraction was least, although its nitrogen content was relatively the highest. The mitochondrial fraction formed only a comparatively small portion of the protein nitrogen of the homogenate. Its content of fluorescent material was also low after hydrolysis, but, per mg. of nitrogen it was only slightly lower than other cytoplasmic fractions. FIG. * This experiment was performed (in the Spring of 1955) with the co-operation of Dr. K. McCarty of the Department of Biochemistry, Columbia University. TABLE I.-Distribution of MDAB in Cytoplasmic Fractions 1.-Protein-bound carcinogen liberated by hydrolysis from separated cytoplasmic fractions of 3 rat livers (43 days of carcinogen feeding). Ethanol-HCl mixture, Vol. 25 ml. FIG. 1.-Protein-bound carcinogen liberated by hydrolysis from separated cytoplasmic fractions of 3 rat livers (43 days of carcinogen feeding). Ethanol-HCl mixture, Vol. 25 ml. When we obtained in another experiment, a mitochondrial fraction which was less contaminated with RNA-rich elements, no fluorescence was found after hydrolysis. , y y These results left little doubt that neither MDAB nor 3,4-benzopyrene are stored in mitochondria, which is in conformity with their failure to produce respiratory inhibition. The mitochondria cannot be thought of as targets for any direct action of these carcinogens. Distribution of MDAB in ribonucleic acid-containing fractions of the cytoplasm The presence of protein-bound carcinogen in the supernatant and ergasto- plasm, fractions containing ribonucleoproteins, and the absence of the carcinogen in mitochondria, free of RNA in any appreciable amount, necessitated investiga- tion of the relationship ofthe carcinogen to nucleoproteins. When liver homogenate from rats fed with carcinogenic azo dye (28 days) was centrifuged at 164000 x g (0.88 M sucrose) for 4 hours, about 95 per cent of all ribonucleoprotein material was spun down. The protein-bound azo dye was, nevertheless, present in the supernatant in a greater amount than in all other fractions combined.* 241 MECHANISM OF CARCINOGENESIS IN LIVER MECHANISM OF CARCINOGENESIS IN LIVER Attempts were also made to isolate, or at least to concentrate, the carcinogen- binding protein from the soluble phase. The initial steps of separation are shown in Table II. Practically all ribonucleoprotein was precipitated by acidification to pH 5-6. The fraction precipitated by the second addition of ammonium sulfate contained most of the protein-bound carcinogen but gave a completely negative pentose test (Mejbaum's modification of Bial's reaction). Ag 210( 180( so1500 1200 900 600 300 7 I VIA I Purified PFFIF0P F=O k / I F 25 ] 20 U)o~ 15 =' E._ C.) C.. N J R S Mitochondria FIG. 2.-Relative proportions of protein nitrogen and protein-bound 3,4-benzpyrene in fractions of mouse epidermis. FIG. 2.-Relative proportions of protein nitrogen and protein-bound 3,4-benzpyrene in fractions of mouse epidermis. TABLE II.-Initial Steps of Purification of the Protein Binding the Azo Dye 1. Livers removed separately and homogenized in cold 0 25 M sucrose. 1. Livers removed separately and homogenized in cold 0 25 M sucrose. 2. Centrifuged, 10 minutes at 13,000 r.p.m. Sediment discarded. 3. Supernatant, 1 M. CH3COOH added to pH 5 8. Centrifuged, sediment discarded. 4 4 4. Supernatant, IN NH4OH added to pH 8.0 + (NH4)2SO4 in powder to concentration 2M. Centrifuged, sediment discarded. . Supernatant + 0-1 M. CH3COOH added to pH 5-4; if sediment forms within 10 minutes, centrifuged and sediment discarded. g 6. Supernatant + (NH4)2SO4 to 3 Moles, centrifuged, reddish supernatant discarded. 6. Supernatant + (NH4)2SO4 to 3 Moles, centrifuged, reddish supernatant discarded. 7 * Sediment dissolved in 7.* Sediment dissolved in N N 1000 100 1000 100 NH4OH in a small volume, dialyzed against HO20 48 hours. NH4OH in a small volume, dialyzed against HO20 48 hours. 8. Crude fraction lyophilized and stored under vacuum in the cold room. This lyophilized fraction is the starting point for isolation of carcinogen-binding protein. * 7 gives a negative pentose test. 8. Crude fraction lyophilized and stored under vacuum in the cold room. lyophilized and stored under vacuum in the cold room. This lyophilized fraction is the starting point for isolation of carcinogen-binding protein. * 7 gives a negative pentose test. The isolated protein fraction had a pale yellow color in solution. Its absorption maximum occurred at 405 mu. (pH 7.0). The absorption maximum is shifted only slightly toward longer wave-lengths in comparison to the absorption maxi- 242 SILVIO FIALA AND ANNA E. FIALA mum of pure carcinocgen dissolved in diluted ethanol and at the same pH (Fig. 3). Normal hydrochloric acid, causing pink coloration, dissolved the protein-dye complex without causing turbidity. It was seen that under these conditions the absorption maximum of the complex was again similar to that of pure carci- nogen. g In the ultraviolet region the undenatured protein fraction containing very low absorption at 260 m/u. had an absorption peak at 280 m/,. The negative pentose test and spectra thus presented convincing evidence that carcinogen-binding protein of the supernatant fraction is not a ribonucleoprotein. p p p In order to establish the relationship of the carcinogen to the ribonucleo- proteins in the ergastoplasm, the lyophilized subfractions (pH 941 soluble sub- A, FIG. 3.-Absorption spectra of carcinogen-binding protein (full line) in ultraviolet and visible region. MECHANISM OF CARCINOGENESIS IN LIVER N/1O NH4C] in N/J0 NH4OH and 0 5 ml. Co (NH3)6C13 (4 x 10-3 M). plasm. Contrary to this expectation the carcinogen was found to be firmly bound mainly to the residuum, apart from ribonucleoproteins, thus providing evidence that it is here where the damaging influence ofthis carcinogen is primarily localized. plasm. Contrary to this expectation the carcinogen was found to be firmly bound mainly to the residuum, apart from ribonucleoproteins, thus providing evidence that it is here where the damaging influence ofthis carcinogen is primarily localized. TABLE II.-Initial Steps of Purification of the Protein Binding the Azo Dye Spectra of crystalline carcinogen (10 tg./ml.) at pH 2-9 and 70. I , FIG. 3.-Absorption spectra of carcinogen-binding protein (full line) in ultraviolet and visible region. Spectra of crystalline carcinogen (10 tg./ml.) at pH 2-9 and 70. fraction and the residuum) were analyzed for ribonucleic acid. It was found that the soluble portion contained practically all the ribonucleoprotein which could be precipitated with CaCl2 and also a protein fraction which did not contain RNA but was relatively rich in polarographically active, protein-bound, sulphydryl groups. The RNA-rich protein fraction could be split off from the soluble sub- fraction with 1 M NaCl. The pH 9.1-insoluble residuum dissolved in N/2 NaOH at room temperature only in the course of several hours, giving a yellow solution. Approximately 1/3 of this residuum dissolved in 0.1 M citric acid (pH 2.5), while the addition of 0.1 M Na citrate produced turbidity. In contrast to the soluble fraction, the insolu- ble residuum was characterized by a low content of polarographically active sulphydryl groups. The amount of sulphydryl groups in the residuum did not increase after heating to 60° for 15 minutes to any considerable degree. It seemed likely that their presence in the residuum was due to the contamination by the soluble fraction. The residuum differed also from the soluble fraction by a relatively low absorption at 280 mp. which belongs to aromatic amino acids. 243 MECHANISM OF CARCINOGENESIS IN LIVER MECHANISM OF CARCINOGENESIS IN LIVER After several hours, the yellow material of the ergastoplasmic residuum dissolved in acidified chloroform. When irradiated in the dark with long ultra- violet light it gave a very intensive green fluorescence. This indicated that the riboflavin (lumiflavin) moiety of yellow enzymes is incorporated into the ergasto- plasmic residuum. Ergastoplasmic residuum from tumors such as Novikoff hepatoma did not yield fluorescence under similar conditions. The properties of the residuum and the fact that it contains few if any sulphydryl groups and is very poor in aromatic amino acids, left no doubt that the residuum represents the insoluble, structural, protein framework of the ergastoplasm, while the RNA- rich material represents Palade granules (Palade, 1955) which are attached from the outside to the tubular network (endoplasmic reticulum, Porter, 1953). (endoplasmic , , 1953). Because of the solubility of carcinogen-binding protein in the supernatant, we expected to find carcinogen localized in the soluble subfraction of the ergasto- FIc. 4.-Comparison of polarographically active free sulphydryl groups in structural (E) and soluble (P) protein fraction of rat liver ergastoplasm. 13 mg. of lyophilized protein fraction dissolved in 5 ml. of 0 5 N NaOH. 0 5 ml. of protein solution added to the mixture of 5 ml. N/1O NH4C] in N/J0 NH4OH and 0 5 ml. Co (NH3)6C13 (4 x 10-3 M). c. 4.-Comparison of polarographically active free sulphydryl groups in structural (E) and soluble (P) protein fraction of rat liver ergastoplasm. FIc. 4.-Comparison of polarographically active free sulphydryl groups in structural (E) and soluble (P) protein fraction of rat liver ergastoplasm. 13 mg. of lyophilized protein fraction dissolved in 5 ml. of 0 5 N NaOH. 0 5 ml. of protein solution added to the mixture of 5 ml. N/1O NH4C] in N/J0 NH4OH and 0 5 ml. Co (NH3)6C13 (4 x 10-3 M). FIc. 4.-Comparison of polarographically active free sulphydryl groups in structural (E) and soluble (P) protein fraction of rat liver ergastoplasm. 13 mg. of lyophilized protein fraction dissolved in 5 ml. of 0 5 N NaOH. 0 5 ml. of protein solution added to the mixture of 5 ml. N/1O NH4C] in N/J0 NH4OH and 0 5 ml. Co (NH3)6C13 (4 x 10-3 M). ( ) p g p 13 mg. of lyophilized protein fraction dissolved in 5 ml. of 0 5 N NaOH. 0 5 ml. of protein solution added to the mixture of 5 ml. 3. The prevention of adaptive formation of tryptophan peroxidase by MDAB All changes described so far as induced by carcinogens are long range effects. By the fact that carcinogenic azo dye MDAB is not a respiratory inhibitor and that glycolysis increases only at the onset of malignant transformation (Fiala and Fiala, 1959) the often held assumption that the carcinogen interferes primarily with the metabolism of the cell cannot be held any longer. What then is the primary effect of a carcinogen beyond its binding to one or more chemically, not yet fully characterized proteins ? 244 SILVIO FIALA AND ANNA E. FIALA It seemed obvious that an answer to this question could be obtained if we could find a reaction normally occurring either in the supernatant or in the ergasto- plasm which would be inhibited by a carcinogen very soon after its incorporation into the cell. We assumed that the depletion of cytoplasmic structures may be due to the interference with some phase of the mechanism of protein synthesis. This possibility could not be excluded by the finding that carcinogen-binding protein is not a nucleoprotein, because the mechanism of protein synthesis is very likely caused by an interplay of several components, as one may judge, e.g. from the experiments of Hoagland, Keller and Zamecnik (1956). One of the best objects in the study of protein synthesis are the adaptive enzymes. It is known that tryptophan peroxidase (TPO) is an adaptive enzyme formed in mammalian liver in increased amounts following the injection of L-tryptophan (Knox, 1951). The enzyme is synthesized de novo from its constituent amino acids (Lee and Williams, 1952). Moreover, it has been reported recently (Claudatus and Ginori, 1957) that there exists a block in the pathway of tryptophan metabolism in hepatoma via kynurenin, indicating that TPO is absent from hepatoma. When we tested the localization of this enzyme in normal rat liver, we found that it is present in the soluble phase. No data seem to exist in the literature on the level of this enzyme and its adaptive synthesis in the precancerous liver. y p y p eca ce ous For those reasons we investigated the induction of TPO in livers of MDAB-fed rats and compared it with controls on a basal diet without carcinogen. We found that the level of TPO did not appreciably change during the first stage of carcino- genesis but that its new formation was largely inhibited by the presence of carci- nogen. DNA mg. in 1 ml. homogenate. Spec. act. -= activity per 5 x 10-12 g. DNA. Activity = micro-moles of Kynurenin formed in 1 ml. homogenate in 1 hour at 37°. DNA = mg. in 1 ml. homogenate. Spec act -= activity per 5 x 10-12 g DNA MECHANISM OF CARCINOGENESIS IN LIVER From these data one can conclude that the carcinogen does indeed inhibit the synthesis of at least some soluble proteins. From these data one can conclude that the carcinogen does indeed inhibit the synthesis of at least some soluble proteins. p In order to decide whether this inhibition is caused by structural damage to the cell, which certainly occurs already during first weeks of carcinogen feeding, or whether it is due to a block of the enzyme forming system by the carcinogen, we applied single massive doses of carcinogen, followed by tryptophan injection and measured TPO activity some six hours later. The whole experiment was performed within twenty-four hours after application of the carcinogen. Table IV shows the result of such experiment. TABLE IV.-Effect of a Single Injection of MDAB on Adaptive Synthesis of Tryptophan Peroxidase r Activity .0.-600 Non-injected DNA . 744 (3) LSpec. act. 0-400 Corn oil Injected rActivity . 1- 300 (Trypt.) - DNA . 608 (3) LSpec. act. 1-060 Increase 0-660, 165% Non-injected DNA .615 . 552 . 550 Corn oil (3) Spec. act. 0-258 . 0 500 . 0- 395 plus carcinogen Activity 0- 615 0- 810 0- 303 Injected DNA . 600 608 . 450 { (3) LSpec. act. . 0-512 . 0-670 . 0-330 Increase . 0-254 . 0-170 . 0 100% ABLE IV.-Effect of a Single Injection of MDAB on Adaptive Synthesis of Tryptophan Peroxidase It is seen that there is a substantial inhibition of TPO formation already after a single injection of the carcinogen so that we may assume with a great degree of probability that it is not structural damage but a block by the carcinogen which is responsible for the inhibition of protein synthesis by the enzyme forming system of the cell. 3. The prevention of adaptive formation of tryptophan peroxidase by MDAB This is shown in Table III. TABLE III.-Effect of MDAB Feeding on Adaptive Synthesis of Tryptophan Peroxidase Time in days A. 0 10 25 30 35 80 Non [Activity 0.420 0 724 0.484 0-287 0-429 0- 640 { on IDNA 702 740 570 544 700 576 Basal injected Spec. act. 0-300 0.489 0-368 0-263 0- 300 0 556 diet Tryptophan rActivity 1.550 1.580 1-240 1-130 1-410 1-672 rACtiVitY 15786 552 500 584 700 524 nJeCte LSpec. act. 0-987 1-436 1-240 0- 966 1 007 1-024 Increase 0 687 0 947 0- 882 0 703 0- 707 1-024 229% 195% 240% 168% 230% 184% No- [ Activity 0-620 0-384 0 366 0.420 0-440 Basal On- DNA - 750 530 764 500 760 diet injected Spec. act. - 0-413 0-359 0-476 0 420 0 290 plus carcinogen Tryptophan Activity - 1-130 0-480 0- 476 0-829 0.550 carinoenTryptophan ~DNA injectedDNA-- 675 530 572 700 692 nJeCte LSpec.act. - 0-836 0-452 0-417 0-440 0-410 Increase - 0-423 0-093 0 0-020 0- 120 100% 25% - - 40% Activity = micro-moles of Kynurenin formed in 1 ml. homogenate in 1 hour at 37°. DNA = mg. in 1 ml. homogenate. Spec. act. -= activity per 5 x 10-12 g. DNA. TABLE III.-Effect of MDAB Feeding on Adaptive Synthesis of Tryptophan Peroxidase TABLE III.-Effect of MDAB Feeding on Adaptive Synthesis of Tryptophan Peroxidase Activity = micro-moles of Kynurenin formed in 1 ml. homogenate in 1 hour at 37°. DNA = mg. in 1 ml. homogenate. mg. in ml. homogenate. Spec. act. -= activity per 5 x 10-12 g. DNA. MECHANISM OF CARCINOGENESIS IN LIVER 245 4. The inhibiting effect of the carcinogen on the ergastoplasm Since the carcinogenic azo dye MDAB is present not only in the soluble phase of the cell, but also in the residuum of the ergastoplasm, the question of how the residuum is affected by the carcinogen led to a problem similar to the one encountered in the case of TPO and the soluble phase of the cell, namely to the choice of a suitable enzyme. Such an enzyme is G 6-Pase, shown to be present exclusively in the microsomes (de Duve et al., 1953). This enzyme was shown to decline in amount during hepatic carcinogenesis and to be absent from hepatoma (Weber and Cantero, 1955). It could be assumed, therefore, that an investigation of the mechanism whereby the carcinogen brings about the depletion of G 6-Pase, would throw some light on the effect of the carcinogen on the ergastoplasm. g g g p The separation of the ergastoplasm into pH 9-1-soluble fraction and the insolu- ble residuum has shown that this enzyme is present exclusively in the residuum. This agrees with the unsuccessful attempts to solubilize the enzyme. g p y When we followed the depletion of G 6-Pase during feeding with M SILVIO FIALA AND ANNA E. FIALA 246 have seen that for a period of several weeks or 2-3 months there is no appreciable change in the level of this enzyme. This agrees with the data of Weber and Cantero (1955) who used DAB as carcinogen. When we correlated the level of this enzyme with the level of anaerobic glycolysis and cellular proliferation we have seen that the period of depletion of G 6-Pase roughly coincides with the sudden increase of anaerobic glycolysis and, within certain limits of error, also with the onset of cellular proliferation. The coincidence of G 6-Pase depletion and of anaerobic glycolysis increase seems to be an important point, which is documented by Table V. E V.-Time-Correlation between G 6-Pase, Increase of Anaerobic Glycolysis and Cellular Proliferation during MDAB Carcinogenesis DNA content of Anaerobic whole liver G 6-Pase glycolysis in 106 units Days activity increase in % (5 X 10-12 g. DNA) Proliferation 32 100 0 3200 . 0 40 . 100 . 0 . 3450 . 0 55 100 . 0 . 2800 . 0 60 100 . 0 . 3100 . 0 75 50 . t40 . 4400 t 76 . 42 t2oo00 8000 . ttt 87 80 . 0 . 4. The inhibiting effect of the carcinogen on the ergastoplasm 3900 0 105 48 . t55 . 4250 . t 150 25 . t195 5200 . tt In further experiments we have seen that single massive doses may decrease the activity of G 6-Pase. However, this effect did not occur regularly and we did not attempt, so far, to further analyze this phenomenon. It seemed more important to study whether the increase in the level of G 6-Pase observed by Weber, Allard, Lamirande and Cantero (1955) induced in the normal liver by several injections of cortisone acetate does occur in the carcinogen storing liver. Under these con- ditions the increase of G 6-Pase seems to increase as an adaptive enzyme. In rats fed with a basal diet and, therefore, under a low protein regimen, the cortisone injection often led to a striking decrease in the weight of liver, instead of increase in the weight usually observed in the normal rats. Such cases were excluded and only these cases were considered in which liver either did not change or increased its weight after 5 consecutive daily injections of 25 mg. of cortisone acetate. This was achieved by placing the animals on regular food during the whole period of experiment. In such cases the increase of G 6-Pase occurred in control, cortisone- injected, animals, but did not occur in carcinogen-treated, cortisone-injected, rats. In order to see whether one or two injections of the carcinogen in normal rats would modify the response of tissue to cortisone in regard to G 6-Pase, the following experiment illustrated in Table VI was performed. g p p The experiment shows that similar to the effect on TPO in the soluble phase, the carcinogen suppresses the increase of an adaptive enzyme of ergastoplasm. The condition, however, may not be strictly analogous since a decrease in G 6-Pase activity was occasionally observed, after single massive doses of carcinogen in distinction to the TPO level which remained unchanged. In any case the carcinogen also exerts an inhibitory effect on the ergastoplasm which is manifested long before structural damage can be observed by differential centrifugation. MECHANISM OF CARCINOGENESIS IN LIVER 247 TABLE VI.-Effect of Cortisone Acetate on the Activity of G 6-Pase of Rat Liver in the Presence of MDAB A (8) B (8) C (8) Cortisone (ug.) (Mg.) (mg.) Controls Non-injected . 10- 6 9 87 . 7- 81 Injected . 23 4 . 1844 . 1165 Increase . 4. The inhibiting effect of the carcinogen on the ergastoplasm - . +120% . +-87% +50% Carcinogen Non-injected . 10-4 93 . 4.15* Injected . 12-66 . 11.23 . 7.77t Increase . - . +22% . +20% . + ? E VI.-Effect of Cortisone Acetate on the Activity of G 6-Pase of Rat Liver in the Presence of MDAB A -= 2 injections of corn oil (10 ml.) with or without carcinogen (60 mg.). j ( ) B = Basal diet to 60 days, with or without carcinogen. C Basal diet for 100 days, with or without carcinogen. Note.-Activity = Ag. inorganic P released in 20 minutes at 37° by tissue homogenate equivalent to 0 05 mg. DNA. * Malignant changes (gross proliferation) in both livers. t i.d. in one liver. to 0 05 mg. DNA. * Malignant changes (gross proliferation) in both livers. t i.d. in one liver. g * Malignant changes (gross proliferation) in both livers. t i.d. in one liver. 5. The effect of MDAB on the structure of the ergastoplasm In the critical stage of liver carcinogenesis the sudden increase of anaerobic glycolysis is linked to the depletion of G 6-Pase. This enzyme has been found (see above) to be anchored in the insoluble residuum of the ergastoplasm while the ribonucleoprotein fraction goes into solution at pH 9 1. This fact led us to investi- gate what happens to the insoluble residuum at this stage of carcinogenesis and we compared, therefore, the quantitative relationship between the residuum and the soluble fraction during carcinogenesis. The results are summarized in Table VII. TABLE VII.-The Proportion between Insoluble (pH 9-0) Residuum (I) and Soluble Proteins (S) in the Ergastoplasm Fraction during Rat Liver Carcinogenesis Novikoff Basal diet MDAB hepatoma Time (days) . . 43 105 . 43 70 87 100 105 119 . - I/S . . . 2-0 1.9 2-0 2-0 1-4 0-66 0.55 0-50 . 04 Total DNA of liver* . 2709 3760 . 3600 3250 3300 4000 4690 5100 - Gross proliferation - - - . - ±- + + . + * Expressed in 106 DNA units (5 x 10-12 g.). TABLE VII.-The Proportion between Insoluble (pH 9-0) Residuum (I) and Proteins (S) in the Ergastoplasm Fraction during Rat Liver Carcinogenes Novikoff Basal diet MDAB hepatoma Time (days) . . 43 105 . 43 70 87 100 105 119 . - I/S . . . 2-0 1.9 2-0 2-0 1-4 0-66 0.55 0-50 . 04 Total DNA of liver* . 2709 3760 . 3600 3250 3300 4000 4690 5100 - Gross proliferation - - - . - ±- + + . + * Expressed in 106 DNA units (5 x 10-12 g.). The table shows that the proportion between the residuum and the soluble fraction does not undergo a noticeable change in the first stage of carcinogenesis. In the second period, however, there occurs a striking change. The same amount of lyophilized material in the control shows a predominance of the residuum, while in the experimental sample and especially in tumor (Novikoff hepatoma) the ratio of the residuum to the soluble fraction is reversed, demonstrating predominance of the soluble fraction and depletion of the residuum, which is the structural groundwork of the ergastoplasm. DISCUSSION In the presence of the carcinogen, the cell becomes increasingly independent of the organism; becoming so to speak a tissue culture within the organism. g From this viewpoint it is probably of utmost importance that the carcinogen is incorporated into the insoluble residuum which constitutes the tubular structures of the ergastoplasm. The system of these structures is known to form a continuous network of membrane-bound cavities, called endoplasmic reticulum (Porter, 1953, Palade, 1955). In distinction to the soluble proteins of the cell, endowed with various enzymatic functions, the insoluble proteins were often considered of secondary importance, merely acting in the role of a cytoskeleton (Needham, 1936, Peters, 1936). It seems, however, more likely that the structural proteins of the cell have a role of utmost importance. Ruska, Edwards and Caesar, (1957) suggested that the tubular network of the endoplasmic reticulum is the conductor of intracellular information and connects the nucleus with the cytoplasm in one system of membranes, including the external cellular membrane. Our experiments have shown that in the critical stage of carcinogenesis there occurs a depletion of this structural protein system of ergastoplasm. It seems very likely that the differences observed in membranes of tumor mitochondria (Mutolo and Abrignani, 1957) are another manifestation of this damage to the structural membrane network of the cell. For this concept, the fact that sudden changes along several parameters (distribution of sulfhydryl groups, cytoplasmic ribonucleic acid, increase of glycolysis, massive proliferation) are coincident with the substantial loss of struc- tural protein from the ergastoplasm is of great importance for the following reasons: It seems plausible to assume that the residual protein of the ergastoplasm is not only the receptor for carcinogen, as we have shown in case of MDAB, but also for tropic hormones. This would agree with the observation that ergastoplasm is the main target for hormones such as ACTH, FSH and TSH (Fiala, Fiala and Sproul, 1957; Fiala, Sproul and Fiala, 1956, 1957). As far as we can judge from early studies of the effect of sex hormones on the epithelial cells of seminal vesicles in the rat (Moore, Hughes and Gallagher, 1930), these hormonal effects are analogous to those exerted by anterior pituitary hormones. DISCUSSION The metabolic damage to the cell induced by a carcinogen such as MDAB is undoubtedly a secondary, long-range effect; a result of whole series of events SILVIO FIALA AND ANNA E. FIALA 248 which are still largely unknown. A more immediate effect than the damage to respiration, is an inhibition of synthesis of certain proteins. The case of the inhibition of TPO formation is an example which is suggestive of harm done by the carcinogen to the ability of the cell to adapt itself to the demands of the organism. At the same time this loss may mean an initial step in progressively increasing independence of the cell from the influence of the environment. This probably represents from a biological viewpoint a gain for the cell which exists on a relatively small rate of protein synthesis when on a basal diet, and is further encroached upon by an additional decrease in protein synthesis in the presence of carcinogen. It has been shown (Westerfeld, Rickert and Hilfinger, 1950) that e.g. xanthine oxidase is considerably depleted in livers of rats on a basal diet and that the carcinogen only accentuates this depletion. Something similar occurs with the levels of respiratory enzymes. In the presence of the carcinogen, the cell becomes increasingly independent of the organism; becoming so to speak a tissue culture within the organism. which are still largely unknown. A more immediate effect than the damage to respiration, is an inhibition of synthesis of certain proteins. The case of the inhibition of TPO formation is an example which is suggestive of harm done by the carcinogen to the ability of the cell to adapt itself to the demands of the organism. At the same time this loss may mean an initial step in progressively increasing independence of the cell from the influence of the environment. This probably represents from a biological viewpoint a gain for the cell which exists on a relatively small rate of protein synthesis when on a basal diet, and is further encroached upon by an additional decrease in protein synthesis in the presence of carcinogen. It has been shown (Westerfeld, Rickert and Hilfinger, 1950) that e.g. xanthine oxidase is considerably depleted in livers of rats on a basal diet and that the carcinogen only accentuates this depletion. Something similar occurs with the levels of respiratory enzymes. DISCUSSION It seems, therefore, that findings pointing to ergastoplasm as the main hormonal target are probably true and in agreement with the concept of Ruska, Edwards and Caesar (1957) concerning the role of the tubular network of the ergastoplasm for the cell: The sudden throwing away of the structural protein would be an adaptive reaction for the cell, allowing it to escape its social duties to the organism by not maintaining synthesis of many 249 MECHANISM OF CARCINOGENESIS IN LIVER proteins, non-essential to its survival, but formed because of essential functions. By throwing this burden away, the cell solves its problem of adaptation. y g y, p p From this viewpoint, several observations are easily understood. The loss of structural protein from the ergastoplasm means disintegration of the tubular network. The RNA-rich granules (Palade-granules) which are attached to the outer surface of these tubules (Palade 1955), are no longer held in an orderly arrangement or in quantitatively sufficient amounts by these tubules. The Palade granules, now randomly distributed, may either disappear or become independent and present in increased amounts within the soluble phase. This explains the observed redistribution of cytoplasmic RNA, reported in the previous paper. Since G 6-Pase is anchored to the residuum, its loss in the critical stage of car- cinogenesis helps in increasing the concentration of glycolytic intermediates. An analogous circumstance seems to be valid for hexosodiphosphatase, indicating that the conditions for increased glycolysis may again be explained by the loss of ergastoplasmic residuum. This represents an additional gain for the cell whose mitochondria became scarce. On the basis of the circumstance that both hormones and carcinogen act on the same structural component and following the concept (Fiala, 1958) that considers the cell as a homeostat, one can understand easily why a hormone may exert, under certain conditions, a carcinogenic effect, as is known for oestrogen (Lacassagne, 1933). The negative feedback in a homeostatic mechanism can easily become positive. It may not be too far fetched to consider the action of certain carcinogenic compounds as an action of "false hormones ". In any case one can draw from our experiments the conclusion that the localization of foreign com- pounds at a vitally important position within, the cell, that is the structural protein of the ergastoplasm, and its ejection by the cell, results in malignant transformation of the cell. SUMMARY The experimental evidence was given to show that carcinogenic azo dye 3'-methyl, 4-dimethylaminoazobenzene is localized in the ergastoplasm and the soluble phase of the cell. The same is true for 3,4 benzopyrene in mouse epidermis. The mitochondria do not contain protein-bound carcinogen. This substantiates the fact that these compounds are not respiratory inhibitors. The carcinogen-binding proteins in the ergastoplasm and in the soluble phase are distinct from nucleoproteins. The ergastoplasm was fractionated into an insoluble residuum, poor in sulphydryl groups and in aromatic aminoacids, and into a soluble fraction. The former contains the carcinogen, the latter ribonucleic acid. The protein-bound carcinogen inhibits new formation of an adaptive liver enzyme, tryptophan peroxidase, and also inhibits the cortisone induced increase of glucose-6-phosphatase even after a single application. g p p The insoluble residuum contains glucose-6-phosphatase. Its disappearance from the cell coincides with increased anaerobic glycolysis. This stage in carcinogenesis is marked by the depletion of the residual structural protein from the ergastoplasm. p g p It was concluded that binding of the carcinogen to this intracellular locus and its consequent irreversible loss means the liberation of the cell from its social 18 250 SILVIO FIALA AND ANNA E. FIALA duties to the organism, this being identical with the malignant transformation of the cell. The senior author (S.F.) wishes to express his thanks to Dr. Edith E. Sproul, Department of Pathology, Columbia University, for her interest in this work. Mr. Harold McQuilla provided valuable assistance in the latter part of this work. This investigation was supported by a grant-in-aid from the National Institutes of Health, U.S. Public Health Service, and also by a grant from the American Cancer Society. Parts of this work were presented in abstract at the Meetings of the American Association for Cancer Research in San Francisco, April 1955 and in Chicago, April 1957. 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https://openalex.org/W3112237773	https://tore.tuhh.de/bitstreams/bd069e97-667a-40f1-a9a0-8219ef9deca3/download	English	null	Characterization and Performance of LbL-Coated Multibore Membranes: Zeta Potential, MWCO, Permeability and Sulfate Rejection	Membranes	2,020	cc-by	11,337	Received: 29 October 2020; Accepted: 1 December 2020; Published: 10 December 2020 Abstract: The characterization of membranes is suitable to investigate changes in the membrane properties caused by Layer-by-Layer (LbL) modiﬁcation. Besides permeability, rejection, and molecular-weight cut-oﬀ(MWCO), which give information about the modiﬁcation of the separation behaviour of the membrane, the zeta potential is capable of describing the surface charge of the membrane and its variation impacted by the properties of the polyelectrolyte multilayers (PEM). In this study, a new method for zeta potential measurement of hollow ﬁbre membranes with several capillaries was developed and further investigations on the LbL modiﬁcation of such membranes were performed. The results showed that an LbL coating with 8 DL PDADMAC/PSS led to a signiﬁcant increase in the membrane charge of more than 20 mV. The coating with a diﬀerent number of polyelectrolyte (PE) layers showed a zig-zag behaviour, comparable to data from ﬂat sheet studies. However, in contrast to most ﬂat sheet membranes, the charge curve assumes a totally negative trajectory at neutral pH. Further experiments on the MWCO of the LbL-modiﬁed membrane showed a reduction in the pore diameter from approx. 20 nm to less than 2 nm, reaching the range of nanoﬁltration membranes. With information on both the zeta potential and the MWCO, it was found that the rejection mechanism in LbL-modiﬁed multibore membranes is a complex interplay between the sieving eﬀect due to reduction in the pore diameter and the repulsion eﬀect of the charged membrane. Keywords: LbL coating; characterization of capillary membranes; zeta potential measurement; MWCO determination; multibore hollow ﬁbre membranes Characterization and Performance of LbL-Coated Multibore Membranes: Zeta Potential, MWCO, Permeability and Sulfate Rejection Saskia Dillmann 1,, Shambhavi Arvind Kaushik 2, Jakob Stumme 2 and Mathias Ernst 1,2 1 Institute for Water Resources and Water Supply, Hamburg University of Technology, Am Schwarzenberg-Campus 3, 20173 Hamburg, Germany; mathias.ernst@tuhh.de 2 DVGW Research Centre TUHH, Am Schwarzenberg-Campus 3, 20173 Hamburg, Germany; shambhavi.kaushik@tuhh.de (S.A.K.); jakob.stumme@tuhh.de (J.S.) Correspondence: saskia.dillmann@tuhh.de Saskia Dillmann 1,, Shambhavi Arvind Kaushik 2, Jakob Stumme 2 and Mathias Ernst 1,2 1 Institute for Water Resources and Water Supply, Hamburg University of Technology, Am Schwarzenberg-Campus 3, 20173 Hamburg, Germany; mathias.ernst@tuhh.de 2 DVGW Research Centre TUHH, Am Schwarzenberg-Campus 3, 20173 Hamburg, Germany; shambhavi.kaushik@tuhh.de (S.A.K.); jakob.stumme@tuhh.de (J.S.) Correspondence: saskia.dillmann@tuhh.de membranes membranes membranes membranes LbL-Modiﬁcation of Membranes Commercial ultraﬁltration (UF) membranes are increasingly being applied in advanced drinking water treatment to remove particles, turbidity, and pathogens. With a pore diameter between 10 and 20 nm, they are generally not suitable for the rejection of dissolved water constituents. Layer-by-Layer surface modiﬁcation can be a powerful tool to modify porous membranes and consequently, their rejection behaviour, in such a way that retentions for certain dissolved water constituents such as sulfate, hardness, or dissolved organic matter can be achieved. During the LbL modiﬁcation, the membrane is alternatingly coated with polycations and polyanions which adsorb on the charged membrane surface and form defect-free, very thin double layers [1]. This process can be Membranes 2020, 10, 412; doi:10.3390/membranes10120412 www.mdpi.com/journal/membranes www.mdpi.com/journal/membranes 2 of 15 Membranes 2020, 10, 412 repeated until the desired layer properties are reached. With increasing layer number, the separation limit of the coated UF membranes successively shifts into the range of nanoﬁltration (NF) membranes [2]. Compared to commercial NF membranes, the LbL-modiﬁed UF membranes might have some advantages: ﬁrstly, the preparation process of LbL-modiﬁed membranes is environmentally friendly when non-toxic polyelectrolytes (PEs) are used [3]. Secondly, the backwash ability of the UF membrane prepared by this process might be maintained [4,5]. Additionally, by controlling the deﬁned coating conditions, it may be possible to tailor the LbL ﬁlm to produce membranes with speciﬁc properties [6,7]. Compared to commercial NF membranes, operation at lower transmembrane pressures (TMP) may be possible as well as direct water treatment, without complex pre-treatment, leading to a more energy-eﬃcient plant operation [8]. The conditions during the membrane coating are decisive for the structure of the polyelectrolyte multilayers and therefore, also for the resulting membrane properties. They can be inﬂuenced by the variation of coating parameters such as the number of double layers (DL), the molecular weight of the PE, temperature, pressure, pH value, or the background ionic strength in the coating solution. In coating solutions with low ionic strength, the PEs are stretched due to higher electrostatic repulsion forces; at high ionic strength, however, they tend to curl up, which leads to the formation of thicker, denser, and more stable layers [9,10]. Therefore, the ionic strength of the coating solution probably inﬂuences the thickness of the polyelectrolyte multilayers (PEMs) to a large extent. The targeted application determines how many double layers are suitable for the LbL modiﬁcation of the membranes. LbL-Modiﬁcation of Membranes For example, Adusumilli and Bruening 2009 used 4–6 double layers of poly(styrenesulfonate) (PSS) and poly(diallyldimethyl ammonium chloride) (PDADMAC) for high SO4−removal (90%) [11], whereas other researchers like Ji et al., 2008 worked with up to 60 bi-layers of polyethyleneimine (PEI) and polyacrylic acid (PAA) on hydrolysed polyacrylonitrile (PAN) ultraﬁltration membranes to achieve separation of alcohol/water mixtures [12]. Certainly, the properties and porosity of the virgin membrane material are also decisive for resulting LbL properties. Another option to inﬂuence the polyelectrolyte multilayer membrane (PEMM) by coating conditions is the “dynamic adsorption” of the PEs [13]. By ﬁltering the polyelectrolyte solution under pressure through the membrane pores, the larger pores are coated ﬁrst and their pore diameters are reduced. This may result in a more uniform pore structure [12,14]. Moreover, coating under pressure can improve the backwash ability as the PEMs seem to be more stable [5]. The deposition of the polyelectrolytes leads to a change in the membrane properties. To determine how the membrane properties change due to the polyelectrolyte coating, common membrane characterization methods can be used, such as molecular weight cut-oﬀ, zeta potential, or contact angle. These methods not only show the change of the membrane properties due to the coating, but also give a hint as to how the ion rejection is inﬂuenced by the modiﬁcation and how coating conditions might be optimized. The MWCO is deﬁned as the molecular weight of a substance that is 90% rejected by the membrane [15]. The MWCO consequently gives information about the pore size of the membrane, as all molecular weights higher than this value have a rejection of at least 90%. The MWCO changes drastically with LbL modiﬁcation of a membrane. For ceramic hollow ﬁbre membranes made from alumina and with a nominal pore size of 100 nm, a coating with six double layers of PDADMAC and PSS led to a MWCO of 240 Da [4]. A similar result of 250 Da was observed for UF hollow ﬁbre PES membranes coated with 8 DL PDADMAC/PSS [2]. The zeta potential gives information on the surface charge of a membrane. As surface charge strongly inﬂuences the rejection of certain water compounds, the zeta potential is a crucial tool for membrane characterization to understand the rejection mechanism in LbL membranes. For LbL-modiﬁed membranes, a general zig-zag pattern is described, depending on the character of the ﬁnal polyelectrolyte layer [11,16–18]. 2.2. Membrane The experiments were carried out on Multibore® membranes (INGE GmbH/Dupont, Greifenberg, Germany), a capillary polyethersulfone (PES) membrane operated inside out. The ultraﬁltration membrane consists of seven single capillaries, each with a diameter of 1.5 mm (MB 1.5) or 0.9 mm (MB 0.9), in one membrane ﬁbre. The capillaries are arranged in a honeycomb structure, therefore having a very high stability. The membrane has an active layer of negatively charged PES and a MWCO of approx. 100,000 Da (manufacturer information). The membranes used in the experiments were lab modules of 25–30 cm, which conforms to a membrane surface of approx. 0.006 m2. The membranes were ﬂushed with at least 1 L deionized water (Milli-Q, Millipore Corporation, Billerica, MA, USA) before coating to remove all residues from the membrane. LbL-Modiﬁcation of Membranes Depending on the virgin membrane material, the polyelectrolyte, and the coating conditions, the zeta potential behaves diﬀerently. PES ﬂat sheet ultraﬁltration membranes, coated with PDADMAC/PSS, showed a tendency progressing towards less negative zeta 3 of 15 Membranes 2020, 10, 412 potential values with an increasing number of layers. Starting at 6 DL, the following layers had a positive zeta potential, independent of if a polycation or -anion was the terminating polyelectrolyte layer [11]. On the contrary, nanoﬁltration membranes, modiﬁed with PDADMAC/PSS, showed a clear shift between positive and negative zeta potential, consistent with the electrostatic orientation of the concluding polyelectrolyte. With the increasing number of layers, the amplitude expands, from about −50 mV at 4 DL to about −70 mV at 8 DL [16]. Su et al., 2012 showed that besides the nature of the virgin membrane and the polyelectrolytes, the coating procedure also plays an important role for the resulting zeta potential [13]. As natural dissolved organic water compounds are generally negatively charged, negatively charged membranes are usually used in water treatment in order to avoid membrane fouling. For LbL-modiﬁed membranes, not only does membrane fouling need to be avoided, but additionally, a negative charge may lead to a higher rejection of anions. In order to examine the charge of the modiﬁed membrane, to describe how it changes with an increasing number of layers, and to draw conclusions on the rejection behaviour of these membranes, it is of major importance to measure the surface charge with a reliable method. To the best of found knowledge, there is no measuring method for zeta potential in hollow ﬁbre membranes with several capillaries (also called multibore membranes from hereon). As cutting the membrane to separate the capillaries would damage the membrane and probably destroy the LbL ﬁlms, a measuring method was developed and tested to analyse the multibore membranes. 2.1. Materials Poly(diallyldimethyl ammonium chloride) (PDADMAC; Mw = 400,000–500,000 g/mol, 20 wt % solution), poly(styrenesulfonate) sodium salt (PSS; Mw = 1,000,000 g/mol, powder), and dextran standards of 70,000, 100,000, and 150,000 Da were purchased from Sigma Aldrich (Taufkirchen, Germany). Sodium chloride (NaCl; Mw = 56.488 g/mol, powder) and magnesium sulfate (MgSO4·7H2O; Mw = 120.366 g/mol, powder) were purchased from Carl Roth (Karlsruhe, Germany). Polyethylene glycol (PEG) standards of 200, 300, 400, 600, 1000, 1500, 2000, 6000, 10,000, 20,000, and 35,000 Da were purchased from Merck KGaA (Darmstadt, Germany). All solutions were prepared in deionized water (Milli-Q, Millipore Corporation, Billerica, MA, USA). 2.3. Coating For the LbL modiﬁcation, the membrane was coated with cationic and anionic polyelectrolytes inside the capillaries, as shown in Figure 1. 4 of 15 4 f 16 4 of 16 Membranes 2020, 10, 412 M b 2020 10 Membranes 2020, 10, x Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). Figure 1. Coating of multibore membrane with varying layers of polyelectrolytes inside the capillaries (image: [19]). The polycation PDADMAC and the polyanion PSS were used for the coating process, both known to be stable against acids, alkalis, and oxidants which are used for the usual cleaning of ultrafiltration membranes [4,20]. PE solutions were prepared in solutions containing a concentration of 0.1 M NaCl background ions. As the membrane is negatively charged, first PDADMAC is deposited on the membrane surface with a coating time of 3 min. After a washing step with pure water to remove the excess PDADMAC, the PSS coating followed also with a contact time of 3 min. The coating with these two PEs is called one double layer and can be repeated as often as necessary to achieve the desired properties. The polycation PDADMAC and the polyanion PSS were used for the coating process, both known to be stable against acids, alkalis, and oxidants which are used for the usual cleaning of ultraﬁltration membranes [4,20]. PE solutions were prepared in solutions containing a concentration of 0.1 M NaCl background ions. As the membrane is negatively charged, ﬁrst PDADMAC is deposited on the membrane surface with a coating time of 3 min. After a washing step with pure water to remove the excess PDADMAC, the PSS coating followed also with a contact time of 3 min. The coating with these two PEs is called one double layer and can be repeated as often as necessary to achieve the desired properties. 2.3. Coating The polycation PDADMAC and the polyanion PSS were used for the coating process, both known to be stable against acids, alkalis, and oxidants which are used for the usual cleaning of ultrafiltration membranes [4,20]. PE solutions were prepared in solutions containing a concentration of 0.1 M NaCl background ions. As the membrane is negatively charged, first PDADMAC is deposited on the membrane surface with a coating time of 3 min. After a washing step with pure water to remove the excess PDADMAC, the PSS coating followed also with a contact time of 3 min. The coating with these two PEs is called one double layer and can be repeated as often as necessary to achieve the desired properties. to achieve the desired properties. For the coating of the ultrafiltration membranes, a coating machine, the Nanocoater (Surflay Nanotec, Berlin, Germany), was used. The coater is computer-controlled to achieve a uniform and reproducible coating on the membrane surface. For the coating of the ultraﬁltration membranes, a coating machine, the Nanocoater (Surﬂay Nanotec, Berlin, Germany), was used. The coater is computer-controlled to achieve a uniform and reproducible coating on the membrane surface. For the coating of the ultrafiltration membranes, a coating machine, the Nanocoater (Surflay Nanotec, Berlin, Germany), was used. The coater is computer-controlled to achieve a uniform and reproducible coating on the membrane surface. reproducible coating on the membrane surface. The lab modules of the capillary membranes (Figure 2b) were directly connected to the Nanocoater. The flat sheet membranes were coated in a self-constructed cell that was also connected to the Nanocoater. The lab modules of the capillary membranes (Figure 2b) were directly connected to the Nanocoater. The ﬂat sheet membranes were coated in a self-constructed cell that was also connected to the Nanocoater. The lab modules of the capillary membranes (Figure 2b) were directly connected to the Nanocoater. The flat sheet membranes were coated in a self-constructed cell that was also connected to the Nanocoater. (a) (b) D D D 1 4 2 3 5 3 3 6 9 8 7 (a) (b) D D D 1 4 2 3 5 3 3 6 9 8 7 (a) (b) Figure 2. Scheme of the filtration plant (a) and the membrane module (b). D D D 1 4 2 3 5 3 3 6 9 8 7 (a) (b) Figure 2. Scheme of the filtration plant (a) and the membrane module (b). 2.3. Coating D D D 1 4 2 3 5 3 3 6 9 8 7 Figure 2. Scheme of the ﬁltration plant (a) and the membrane module (b). (b) (b) (a) (a) ( ) ( ) Figure 2. Scheme of the filtration plant (a) and the membrane module (b). Figure 2. Scheme of the filtration plant (a) and the membrane module (b). Figure 2. Scheme of the ﬁltration plant (a) and the membrane module (b). 2 4 Filtration Setup and Experiments 2.4. Filtration Setup and Experiments 2.4. Filtration Setup and Experiments 2.4. Filtration Setup and Experiments The filtration experiments were performed with a lab-scale filtration plant (Figure 2a) for capillary membranes (Figure 2b). The membrane (4) was potted in an acrylic pipe with a feed inlet, a The filtration experiments were performed with a lab-scale filtration plant (Figure 2a) for capillary membranes (Figure 2b). The membrane (4) was potted in an acrylic pipe with a feed inlet, a The ﬁltration experiments were performed with a lab-scale ﬁltration plant (Figure 2a) for capillary membranes (Figure 2b). The membrane (4) was potted in an acrylic pipe with a feed inlet, a permeate, 5 of 15 Membranes 2020, 10, 412 and a retentate outlet so that it could be operated in dead-end as well as cross-ﬂow mode. The permeate was collected on a scale (5, 6), so the ﬂux and permeability could be determined. Three pressure gauges (3) were placed to determine the TMP and a control valve (7) was used to regulate the stream, the recirculation, and the TMP. All experiments were implemented in cross-ﬂow mode at a cross-ﬂow velocity of 0.03 m/s and a yield of 25%. Depending on the layer number, the plant was operated at an increasing TMP, starting at 0.2 bar at layer no. 1 up to a maximum pressure of 2.5 bar for 5–8 DL. For ﬁltration experiments regarding the rejection behaviour of the membranes, a 100 mg/L sulfate solution of magnesium sulfate was used. For the determination of the MWCO, 1 g/L solutions of PEG and dextran standards were used. 2.6. MWCO The molecular weight cut-oﬀis one of the most commonly used methods for membrane characterization. According to Crittenden et al., 2012 [24], the pore diameter can be calculated by Equation (1) using the MWCO. dpore = 0.11·M0.46 (1) (1) dpore = 0.11·M0.46 dpore = hydraulic pore diameter, nm; M = molecular weight according to MWCO, g/mol. The determination of the MWCO was done with polyethylene glycol (PEG) and dextran standards of diﬀerent molecular weights, which were used in a mixture of 1 g/L for each substance as the feed solution. The feed and permeate samples were analysed with gel permeation chromatography with an RI detector. 2.5. Characterization Methods In addition to the pure water permeability and sulfate rejection rates, membrane material properties such as the molecular weight cut-oﬀ[21] and the zeta potential were determined [22]. Moreover, scanning electron microscope (SEM) pictures were taken to show the morphology of the membranes. The permeability and rejection were determined with MB1.5 membranes. Three membranes were coated with double layers of PDADMAC/PSS. Subsequently, the permeability and sulfate rejection was tested in the ﬁltration plant. The separation behaviour of the modiﬁed membranes was tested by the determination of the rejection of divalent ions, SO42−from MgSO4·7H2O. The sulfate concentration in the feed and permeate was measured using the photometric measurement after the precipitation reaction with barium chloride (referring to the Hach Lange Method 8051 [23]). 2.7. SEM Imaging The SEM pictures were taken at the “central service unit SEM” of the Hamburg University of Technology. The membrane samples were sputtered with a 5 nm gold layer and analysed in the scanning electron microscope Zeiss Supra 55 VP FEG-SEM. 3.1. Zeta Potential—Innovative Measurement Method for Multibore Membranes The zeta potential can be used to describe the status of the charge of surf More precisely, the length and the diameter of the cell had to be adapted, as several trials showed that a too short or even a too long piece of membrane led to a termination in the measurement. Several trials revealed that only at a length of 8 cm, it was possible to reach a pressure of 200 mbar in uniform gradual steps and ensure a successful measurement. A dditi l bl d f th t d d fi t 2 DL t d ltib b An additional problem occurred for the uncoated and ﬁrst 2 DL coated multibore membranes. The pressure of 200 mbar already led to a permeation of the solution through the membrane. After a certain time of measurement, the pressure in the system decreased due to the solution permeation, leading to a pressure loss and in consequence, to a termination of the whole measurement. To overcome this challenge, various options of sealing the outside surface of the membrane were tested. The use of diﬀerent glues as well as shrinking tubes were tried in vain, as the measurement was either aborted or did not show a constant and reliable result. In the end, a tube was used whose inner diameter conforms to the outer diameter of the membrane. Two ﬂexible pipes were used to connect the membrane to the electrodes. These adjustments prevented leakages and a stable and plausible measurement was achieved. A scheme of the new constructed setup is shown in Figure 3. An additional problem occurred for the uncoated and first 2 DL coated multibore membranes. The pressure of 200 mbar already led to a permeation of the solution through the membrane. After a certain time of measurement, the pressure in the system decreased due to the solution permeation, leading to a pressure loss and in consequence, to a termination of the whole measurement. To overcome this challenge, various options of sealing the outside surface of the membrane were tested. The use of different glues as well as shrinking tubes were tried in vain, as the measurement was either aborted or did not show a constant and reliable result. In the end, a tube was used whose inner diameter conforms to the outer diameter of the membrane. Two flexible pipes were used to connect the membrane to the electrodes. These adjustments prevented leakages and a stable and plausible measurement was achieved. 3.1. Zeta Potential—Innovative Measurement Method for Multibore Membranes The zeta potential can be used to describe the status of the charge of surf The zeta potential can be used to describe the status of the charge of surfaces, including uncoated and PDADMAC/PSS-modiﬁed membranes. In hollow ﬁbre membranes with several capillaries, the measurement is diﬃcult as the measurement device is usually designed for the determination of single hollow ﬁbres with a limited outer diameter [25]. The major limitation for the multibore system was the achievement of a pressure gradient that is necessary for the zeta measurement. While ﬂat sheet membranes are measured in a cell with a 100 µm gap between the membranes, single bore membranes cannot have an outer diameter larger than 2 mm since the needed pressure gradient cannot be reached and the measurement is then aborted. For the multibore membrane, which has an outer diameter of 4 mm, a suﬃcient pressure gradient was needed over all seven capillaries of the membrane. and PDADMAC/PSS-modified membranes. In hollow fibre membranes with several capillaries, the measurement is difficult as the measurement device is usually designed for the determination of single hollow fibres with a limited outer diameter [25]. The major limitation for the multibore system was the achievement of a pressure gradient that is necessary for the zeta measurement. While flat sheet membranes are measured in a cell with a 100 µm gap between the membranes, single bore membranes cannot have an outer diameter larger than 2 mm since the needed pressure gradient cannot be reached and the measurement is then aborted. For the multibore membrane, which has an outer diameter of 4 mm, a sufficient pressure gradient was needed over all seven capillaries of the membrane. It was thus required to make physical changes to the measuring cell for single bore membranes It was thus required to make physical changes to the measuring cell for single bore membranes. More precisely, the length and the diameter of the cell had to be adapted, as several trials showed that a too short or even a too long piece of membrane led to a termination in the measurement. Several trials revealed that only at a length of 8 cm, it was possible to reach a pressure of 200 mbar in uniform gradual steps and ensure a successful measurement. It was thus required to make physical changes to the measuring cell for single bore membranes. 3.1. Zeta Potential—Innovative Measurement Method for Multibore Membranes The zeta potential can be used to describe the status of the charge of surf A scheme of the new constructed setup is shown in Figure 3. Figure 3. Experimental setup for measuring zeta potential of multibore membranes. Figure 3. Experimental setup for measuring zeta potential of multibore membranes. igure 3. Experimental setup for measuring zeta potential of multibore membranes. igure 3. Experimental setup for measuring zeta potential of multibore membranes. With these fresh adaptations to the measuring system, measurements of the zeta potential in multibore membranes were successfully performed With these fresh adaptations to the measuring system, measurements of the zeta potential in multibore membranes were successfully performed. 2.8. Zeta Potential—Measurement of Flat Sheet Membranes The zeta potential, which arises as an electrical double layer developed at the solid–liquid interface, describes the surface charge of a membrane. It is inﬂuenced by the functional groups on the membrane surface, the ions in the feed solution, and the pH. The zeta potential for capillaries can be described by an approximation of the Helmholtz–Smoluchowski equation [22]. For zeta potential measurements, a SurPASS Electrokinetic Analyzer (Anton Paar GmbH, Graz, Austria) was used and adapted. The ﬂat sheet membranes were prepared for the measurement by storing it in pure water for 24 h to compensate the swelling of the membrane and to remove residual substances. It was then ﬁxed on the stamps of the measuring cell and connected with the measuring electrodes. The gap between the two membranes was adjusted to 100 µm in order to be able to apply the needed pressure gradient. The measurement was performed at 1 mmol/L KCl, starting at pH 9. 6 of 15 6 of 16 Membranes 2020, 10, 412 Membranes 2020, 10, x After the cell was washed by the electrolyte, the zeta potential was determined at diﬀerent pH values between 9 and 3, by titration of HCl. The zeta potential was calculated with the software Attract (supplied by Anton Paar) based on the Helmholtz–Smoluchowski equation. To compare the results, the capillary and ﬂat sheet membranes both were measured by the Streaming Potential method. between 9 and 3, by titration of HCl. The zeta potential was calculated with the software Attract (supplied by Anton Paar) based on the Helmholtz–Smoluchowski equation. To compare the results, the capillary and flat sheet membranes both were measured by the Streaming Potential method. 3. Results and Discussion After the cell was washed by the electrolyte, the zeta potential was determined at diﬀerent pH values between 9 and 3, by titration of HCl. The zeta potential was calculated with the software Attract (supplied by Anton Paar) based on the Helmholtz–Smoluchowski equation. To compare the results, the capillary and ﬂat sheet membranes both were measured by the Streaming Potential method. between 9 and 3, by titration of HCl. The zeta potential was calculated with the software Attract (supplied by Anton Paar) based on the Helmholtz–Smoluchowski equation. To compare the results, the capillary and flat sheet membranes both were measured by the Streaming Potential method. 3. Results and Discussion 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes To validate the zeta potential measurements, values of the multibore membranes were compared with the results of the flat sheet PES membrane UP 150 from Microdyn-Nadir (Wiesbaden, Germany) Due to the same material (PES) and similar MWCO/pore size situation (approx 150 kDa) To validate the zeta potential measurements, values of the multibore membranes were compared with the results of the ﬂat sheet PES membrane UP 150 from Microdyn-Nadir (Wiesbaden, Germany). Due to the same material (PES) and similar MWCO/pore size situation (approx. 150 kDa), a comparable 7 of 15 Membranes 2020, 10, 412 zeta potential of both membranes was expected. Results of the measurements are displayed in Figure 4. The graph of the non-coated multibore membrane is in very good agreement with that of the ﬂat sheet membrane. Both PES membranes exhibit a zeta potential of approx. −18 mV at a pH of 3, decreasing similarly with increasing pH. The results were quite reproducible, leading to the conclusion that the adaption of the method to measure the zeta potential of hollow ﬁbre membranes with several capillaries was successfully adapted. displayed in Figure 4. The graph of the non-coated multibore membrane is in very good agreement with that of the flat sheet membrane. Both PES membranes exhibit a zeta potential of approx. −18 mV at a pH of 3, decreasing similarly with increasing pH. The results were quite reproducible, leading to the conclusion that the adaption of the method to measure the zeta potential of hollow fibre membranes with several capillaries was successfully adapted. (a) (b) Figure 4. Zeta potential measurement for uncoated (a) and 8 DL-coated (b) capillary and flat sheet membranes. -45 -35 -25 -15 -5 5 15 2 3 4 5 6 7 8 9 Zetapotential [mV] pH Flat sheet membrane, uncoated Capillary membrane, uncoated -45 -35 -25 -15 -5 5 15 2 3 4 5 6 7 8 9 Zetapotential [mV] pH Flat sheet membrane, coated (8 DL) Capillary membrane, coated (8 DL) Figure 4. Zeta potential measurement for uncoated (a) and 8 DL-coated (b) capillary and ﬂat sheet membranes. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes (a) -45 -35 -25 -15 -5 5 15 2 3 4 5 6 7 8 9 Zetapotential [mV] pH Flat sheet membrane, uncoated Capillary membrane, uncoated (b) -45 -35 -25 -15 -5 5 15 2 3 4 5 6 7 8 9 Zetapotential [mV] pH Flat sheet membrane, coated (8 DL) Capillary membrane, coated (8 DL) (b) (a) Figure 4. Zeta potential measurement for uncoated (a) and 8 DL-coated (b) capillary and flat shee membranes. Figure 4. Zeta potential measurement for uncoated (a) and 8 DL-coated (b) capillary and ﬂat sheet membranes. The results of the zeta potential measurement of the membranes coated with 8 DL o PDADMAC/PSS are also in good agreement (Figure 4b). The coated flat sheet as well as multibor membrane have a negative zeta potential of approx. −18 mV at pH 8.5, which then decreases (become less negative) with decreasing pH. For the multibore membrane, the isoelectric point (IEP) is reached at pH 3, while the flat sheet membrane reaches it at a pH of 4.5. These discrepancies in IEPs might be due to the effect of layer defects due to an inhomogeneous coating in the self-constructed test-cell The inflow of the PE solution into the planar coating cell through a single tube may not have fully ensured plug flow conditions of the coating solutions and might have resulted in non-idea homogeneous coating over the membrane surface. The results of the zeta potential measurement of the membranes coated with 8 DL of PDADMAC/PSS are also in good agreement (Figure 4b). The coated ﬂat sheet as well as multibore membrane have a negative zeta potential of approx. −18 mV at pH 8.5, which then decreases (becomes less negative) with decreasing pH. For the multibore membrane, the isoelectric point (IEP) is reached at pH 3, while the ﬂat sheet membrane reaches it at a pH of 4.5. These discrepancies in IEPs might be due to the eﬀect of layer defects due to an inhomogeneous coating in the self-constructed test-cell. The inﬂow of the PE solution into the planar coating cell through a single tube may not have fully ensured plug ﬂow conditions of the coating solutions and might have resulted in non-ideal homogeneous coating over the membrane surface. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes g g After the successful adaptation of the measuring method, the effect of the LbL coating on the multibore membranes with regard to the zeta potential was investigated in more detail (Figure 5). After the successful adaptation of the measuring method, the eﬀect of the LbL coating on the multibore membranes with regard to the zeta potential was investigated in more detail (Figure 5). g p g g The zeta potential of the virgin PES multibore membrane is negative over the total pH range: −43 mV at pH 9 and −16 mV at pH 3, due to the negative charge of the PES material of the active membrane layer. After coating with 8 DL of PDADMAC/PSS, it shifts to less negative values, starting at −17 mV at pH 9 and reaching the isoelectric point at pH 3.1. One would expect that the membrane coated with negatively charged PSS as the terminating layer would have a more negative zeta potential. The increasing zeta potential with decreasing pH is attributed to the dissociation of acidic functional groups—at lower pH, the concentration of H3O+ becomes more dominant and the surface starts assuming a positive charge [23]. However, here, the shift in zeta potential to less negative values for 8 DL membranes as compared to the virgin membrane was, in fact, a consequence of the increased number of PE layers. For the 8.5 DL coated membrane with the positively charged PDADMAC as the top layer, it was expected that the zeta potential would be rather positive over the entire pH range. As shown in Figure 5, however, the zeta potential starts with a negative value of −11 mV at pH 9, reaches the IEP at a pH of 6.7, and increases up to 16 mV at pH 3. Although the zeta potential is positive below pH 6.7, it may be possible that at higher pH, the inﬂuence of the negative charge of the membrane is stronger and therefore, partly shields the positively charged functional groups of PDADMAC [13]. 8 of 15 Membranes 2020, 10, 412 Membranes 2020, 10, x 8 of 16 Figure 5. Zeta potential of uncoated (dashed), 8 DL- (line), and 8.5 DL- (dotted) coated membranes. -45 -35 -25 -15 -5 5 15 25 3 4 5 6 7 8 9 Zeta potential [mV] pH Capillary membrane, coated (8 DL) Capillary membrane, uncoated Capillary membrane, coated (8.5 DL) Figure 5. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes Zeta potential of uncoated (dashed), 8 DL- (line), and 8.5 DL- (dotted) coated membranes. Figure 5. Zeta potential of uncoated (dashed), 8 DL- (line), and 8.5 DL- (dotted) coated membranes. Figure 5. Zeta potential of uncoated (dashed), 8 DL- (line), and 8.5 DL- (dotted) coated membranes. The zeta potential of the virgin PES multibore membrane is negative over the total pH range: −43 mV at pH 9 and −16 mV at pH 3, due to the negative charge of the PES material of the active membrane layer. After coating with 8 DL of PDADMAC/PSS, it shifts to less negative values, starting at −17 mV at pH 9 and reaching the isoelectric point at pH 3.1. One would expect that the membrane coated with negatively charged PSS as the terminating layer would have a more negative zeta potential. The increasing zeta potential with decreasing pH is attributed to the dissociation of acidic In order to obtain a better understanding of the electrostatic behaviour of the virgin multibore membrane and how it changes by polyelectrolyte coating, the development of zeta potential with each layer of PES was investigated. The zeta potential of the PSS terminated layers was expected to be negative as it is a negatively charged polyelectrolyte. Conversely, it was expected that the PDADMAC terminated layer would have a rather positive zeta potential due to the positively charged PE. In Figure 6, the values for zeta potential at ca. pH 7 for every single layer are displayed. Membranes 2020, 10, x 9 of 16 functional groups at lower pH, the concentration of H3O becomes more dominant and the surface starts assuming a positive charge [23]. However, here, the shift in zeta potential to less negative values for 8 DL membranes as compared to the virgin membrane was, in fact, a consequence of the increased number of PE layers. For the 8.5 DL coated membrane with the positively charged PDADMAC as the top layer, it was expected that the zeta potential would be rather positive over the entire pH range. As shown in Figure 5, however, the zeta potential starts with a negative value of −11 mV at pH 9, reaches the IEP at a pH of 6.7, and increases up to 16 mV at pH 3. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes 9 of 15 Membranes 2020, 10, 412 In layer numbers 1–4, it can be seen that the PSS terminated layers are in the range of the virgin membrane or even have a more negative value (e.g., −36 mV at 2 DL and −54 mV at 3 DL). This eﬀect can be attributed to the contribution of the negative charge of the polyelectrolyte. From layer number 5, the more positive/less negative trend of the zeta potential is apparent, as the zeta potential reaches a value of −16 mV at 8 DL. Compared to the PSS terminated layers, zeta potential values for the PDADMAC terminated layers were expected to be positive due to the positively charged PDADMAC as the uppermost layer. However, as displayed in Figure 6, in a neutral pH range of pH 7, the membrane possessed a negative zeta potential over all layers. This eﬀect can be explained by the eﬀect that the negative charge of the membrane and negatively charged PSS are partly shielding the positively charged amino groups of the PDADMAC, leading to a total negative zeta potential [13]. Only in the acidic range, at pH 3, the zeta potential became positive after layer number 5.5 and showed increasingly positive potentials for the following layers. g y Despite the negative zeta potential at a pH of 7 over all layers, an increasing trend in the zeta values can be seen. The zeta potential shifts to a less negative range from −67 mV at 1 DL to −16 mV at 8 DL, as more layers are deposited on the membrane. The zig-zag eﬀect is known from the literature, e.g., reported by Reurink et al., 2018 [18] in hollow ﬁbre membranes as well as Adusumilli and Bruening 2009 [11], where PES ﬂat sheet membranes were coated with PSS and PDADMAC. The zeta potential of the membranes was positive for all PDADMAC terminated layers and also became positive even for PSS terminated layers, after the deposition of 6 DL. With further experimental methods, Adusumilli and Bruening found that a higher amount of PDADMAC enters the layers, compared to PSS, and tends to migrate to the surface. Thus, after a certain number of layers, when PSS is further deposited, PDADMAC forms complexes with the PSS, yielding a more positive potential [11]. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes Although the zeta potential is positive below pH 6.7, it may be possible that at higher pH, the influence of the negative charge of the membrane is stronger and therefore, partly shields the positively charged functional groups of PDADMAC [13]. In order to obtain a better understanding of the electrostatic behaviour of the virgin multibore membrane and how it changes by polyelectrolyte coating, the development of zeta potential with each layer of PES was investigated. The zeta potential of the PSS terminated layers was expected to be negative as it is a negatively charged polyelectrolyte. Conversely, it was expected that the PDADMAC terminated layer would have a rather positive zeta potential due to the positively charged PE. In Figure 6, the values for zeta potential at ca. pH 7 for every single layer are displayed. Figure 6. Zeta potential measurements of each layer of LbL-modified multibore membrane. -70 -60 -50 -40 -30 -20 -10 0 0 1 2 3 4 5 6 7 8 9 Zeta potential [mV] Layer number Zeta potential at pH_7 Figure 6. Zeta potential measurements of each layer of LbL-modiﬁed multibore membrane. PE. In Figure 6, the values for zeta potential at ca. pH 7 for every single layer are dis Figure 6. Zeta potential measurements of each layer of LbL-modified multibore membrane. Figure 6. Zeta potential measurements of each layer of LbL-modiﬁed multibore membrane. The curve behaves in a zig-zag pattern, depending on the terminating layer, which is a known effect from the literature data of flat sheet membranes [11,16,17]. The PSS terminating layers have, in general, a more negative value compared to the PDADMAC terminated layers. PDAPMAC terminated layers exhibit a higher zeta potential (less negative) albeit with a negative sign over the h l H I l b 1 4 i b h h PSS i d l i h The curve behaves in a zig-zag pattern, depending on the terminating layer, which is a known eﬀect from the literature data of ﬂat sheet membranes [11,16,17]. The PSS terminating layers have, in general, a more negative value compared to the PDADMAC terminated layers. PDAPMAC terminated layers exhibit a higher zeta potential (less negative) albeit with a negative sign over the whole pH range. 3 2 Zeta Potential of Multibore Membranes 3.2. Zeta Potential of Multibore Membranes The contrary eﬀect was observed by Malaisamy et al., 2011 for PDADMAC/PSS-modiﬁed NF270 membranes—the zig-zag variation of the zeta potential with each layer increases in magnitude as the number of layers increases; i.e., PSS terminated membranes are more negative and PDADMAC terminated membranes are more positive at higher number of layers [16]. However, diﬀerences in the coating procedure as well as the membrane geometry may also inﬂuence the zeta potential. As described in Section 2.3, the multibore membranes in this study were coated with the Nanocoater (Surﬂay Nanotec, Berlin, Germany), which ensures a proper and deﬁned coating of capillary membranes and its active separation layer. It is likely that the charge of the supporting structure also has an impact on the zeta potential, as the surface is comparably larger than the surface of the active layer. It is possible that the negative charge of the supporting structure of the multibore membranes used in this work inﬂuences the measurement, leading to the same zig-zag pattern as known in the literature, but with a shift to the negative range. Additionally, speciﬁcally in the case of multibore membranes, the supporting structure of one capillary is not the only one inﬂuencing the charge; rather, the whole supporting structure around all the seven capillaries contributes to the overall charging behaviour of the membrane. Su et al. 2012 showed that diﬀerent methods in depositing the polyelectrolyte layers on a polysulfone (PS) membrane result in very diﬀerent zeta potential results. When the membrane was coated in a static assembly, one layer of PDADMAC resulted in a negative zeta potential, due to the stronger negative charge of the PS membrane, shielding some of the positively charged amino groups of the PDADMAC. In contrast, with a dynamic assembly, ﬁltering the polyelectrolyte solution in cross-ﬂow mode through the membrane, and thereby also coating the support structure of the membrane, the zeta potential became +55 mV [13]. These ﬁndings support the theory that the zeta potential is strongly impacted by the charge of the support layer and the overall coating procedure. 3.3. Inﬂuence of Zeta Potential on the Permeability and Rejection 3.3. Inﬂuence of Zeta Potential on the Permeability and Rejection As shown in the previous section, the surface charge of the membrane changes drastically due to the LbL coating. The inﬂuence of zeta potential on the normalized ﬂux (ﬂux normalized to one bar) 10 of 15 one bar) tandard Membranes 2020, 10, 412 to the LbL coating. T and sulfate rejection and sulfate rejection is displayed in Figure 7. The mean values of three membranes and the standard deviation are shown for the development of the normalized ﬂux and sulfate rejection, with increasing number of layers. p j , g number of layers. According to the manufacturer, the pure water permeability of the membrane is 1000 L∙m−2∙h−1∙bar−1. In our filtration plant, a pure water permeability of 720 L∙m−2∙h−1∙bar−1 was reached. Figure 7. Normalized flux, sulfate rejection, and zeta potential of LbL-modified multibore membranes vs. number of PE layers (PDADMAC/PSS). 0 1 10 100 1000 -70 -20 30 80 130 180 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 Rejection [%], Normalized Flux [L∙m-2∙h-1∙bar-1] Zeta potential [mV] Layer number Rejection [%] Normalized Flux [L∙m-2∙h-1∙bar-1] Zeta potential [mV] [L∙m-2∙h-1∙bar-1] Figure 7. Normalized ﬂux, sulfate rejection, and zeta potential of LbL-modiﬁed multibore membranes vs. number of PE layers (PDADMAC/PSS). Figure 7. Normalized flux, sulfate rejection, and zeta potential of LbL-modified multibore membranes vs. number of PE layers (PDADMAC/PSS). Figure 7. Normalized ﬂux, sulfate rejection, and zeta potential of LbL-modiﬁed multibore membranes vs. number of PE layers (PDADMAC/PSS). The data of the normalized flux as well as the rejection show a zig-zag pattern in accordance with the data of the zeta potential. In general, the flux, starting with more than 700 L∙m−2∙h−1∙bar−1 fo According to the manufacturer, the pure water permeability of the membrane is 1000 L·m−2·h−1·bar−1 In our filtration plant, a pure water permeability of 720 L·m−2·h−1·bar−1 was reached. The data of the normalized flux as well as the rejection show a zig-zag pattern in accordance with the data of the zeta potential. In general, the flux, starting with more than 700 L∙m−2∙h−1∙bar−1 for According to the manufacturer, the pure water permeability of the membrane is 1000 L·m−2·h−1·bar−1. In our filtration plant, a pure water permeability of 720 L·m−2·h−1·bar−1 was reached. with the data of the zeta potential. 3.3. Inﬂuence of Zeta Potential on the Permeability and Rejection In general, the flux, starting with more than 700 L m h bar for the uncoated membrane, diminishes after coating of the first two layers by nearly 95% to a value of 40 L∙m−2∙h−1∙bar−1. In the following six layers, the decrease is less severe, but nevertheless, it decreases continuously until it reaches a minimum value of approx. 12 L∙m−2∙h−1∙bar−1 at 8 DL. Simultaneously, The data of the normalized ﬂux as well as the rejection show a zig-zag pattern in accordance with the data of the zeta potential. In general, the ﬂux, starting with more than 700 L·m−2·h−1·bar−1 for the uncoated membrane, diminishes after coating of the ﬁrst two layers by nearly 95% to a value of 40 L·m−2·h−1·bar−1. In the following six layers, the decrease is less severe, but nevertheless, it decreases continuously until it reaches a minimum value of approx. 12 L·m−2·h−1·bar−1 at 8 DL. Simultaneously, along with the normalized ﬂux, the rejection of the divalent ion, sulfate, increases nearly linearly with each DL (PSS terminating) up to 70% at 8 DL. It is apparent that the PEMs retain the sulfate ions, but simultaneously cause an additional hydraulic membrane resistance. This is in agreement with the literature data [26,27] of ﬂat sheet and hollow ﬁbre membranes, where, depending on the coating conditions, approx. 75% [27] and up to 96% [26] sulfate rejection were reached. De Grooth et al. 2015 also explained the development of permeability by two diﬀerent eﬀect—the pore-dominating and the layer-dominating regime [26]. Due to a drastic drop in the permeability, it seems apparent that the coating of the membrane starts with attachment of polyelectrolytes in the membrane pores in the ﬁrst two layers, also called the pore-dominated regime, leading to an abrupt decline in the permeability. Thereafter, it is probable that the coating continues on top of the membrane pore structure, called the layer-dominated regime, leading to a less steep decline in the permeability. It is likely that the PEMs form an additional, nearly dense membrane on top of the underlying UF membrane, which causes a further decrease in the permeability and the continuous increase in rejection, subsequently leading to membrane properties that correspond to NF membranes [21]. Even though the PE layers on the membrane are not cross-linked, initial lab scale experiments with the 8 DL-coated membrane show a good stability during backwash. 3.3. Inﬂuence of Zeta Potential on the Permeability and Rejection The permeability stayed nearly constant over 10 backwash and ﬁltration cycles at a backwash pressure of 2.5 bar for 60 s (data not shown). Pilot experiments in a 11 of 15 Membranes 2020, 10, 412 waterworks are planned to investigate PEM long-term stability under higher cross-ﬂow velocities, diﬀerent backwash ﬂuxes, and use of chemical cleaning agents. The PDADMAC terminated layers generally show higher permeabilities compared to the previous PSS terminated layer. It is known from the literature [10,28,29] that PDADMAC has a much higher swelling behaviour than PSS. The polymer ﬁlm swells more when PDADMAC is the terminating layer, leading to thicker but less dense layers. This swelling eﬀect may lead to a higher membrane permeability [13]. Additionally, McCormick et al. [30] observed that water is more mobile within the PE ﬁlm in PDADMAC terminated membranes than in PSS terminated membranes. The reason for this is still not solved. As for the phenomenon of the simultaneous lower rejection of sulfate for the PDADMAC terminated membranes, there are two possible explanations. On the one hand, it is possible that the rejection is constant, but due to dilution, because of the higher permeability, the average rejection drops. On the other hand, the loose structure of the layers may lead to lower rejection rates for sulfate ions compared to the membranes with the PSS terminated coating. Furthermore, it can be suggested that repulsive forces of the negatively charged PSS layer and the SO42−ion are responsible for the higher rejection of the PSS terminating membranes. 3.4. SEM Analysis of Coated and Uncoated Membranes SEM pictures of the membrane give additional information on how the morphology of the membrane is changed by the polyelectrolyte coating. In the following ﬁgure, pictures of the top view of the uncoated and 8 DL-coated multibore membrane are presented (Figure 8). Membranes 2020, 10, x 12 of 16 Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Figure 8. SEM pictures of uncoated (a) and 8 DL-coated (b) multibore membranes. Top view pictures of the membranes are considered in high resolution to show the effect of the coating on top of the membrane surface. In Figure 8a, the uncoated membrane can be seen with pores in a range of 10–30 nm; after LbL coating with 8 DL (Figure 8b), the surface becomes smooth and no pores are visible anymore. This effect supports the above-mentioned theory that the PEMs form an additional dense structure on top of the underlying membrane. Top view pictures of the membranes are considered in high resolution to show the eﬀect of the coating on top of the membrane surface. In Figure 8a, the uncoated membrane can be seen with pores in a range of 10–30 nm; after LbL coating with 8 DL (Figure 8b), the surface becomes smooth and no pores are visible anymore. This eﬀect supports the above-mentioned theory that the PEMs form an additional dense structure on top of the underlying membrane. 3.5. Determination of the Molecular Weight Cut-Off 3.5. Determination of the Molecular Weight Cut-Oﬀ f g ff To determine the MWCO, the rejection of different molecular weight substances that were filtered through the membrane was measured. Consequently, based on PEG and dextran rejection, it was possible to obtain an MWCO curve for the uncoated and coated membranes. According to the manufacturer information (Inge GmbH/Dupont, Greifenberg, Germany), the MWCO of uncoated Multibore membranes should be at 100,000 Da. Investigating the MWCO value based on the To determine the MWCO, the rejection of diﬀerent molecular weight substances that were ﬁltered through the membrane was measured. Consequently, based on PEG and dextran rejection, it was possible to obtain an MWCO curve for the uncoated and coated membranes. According to the manufacturer information (Inge GmbH/Dupont, Greifenberg, Germany), the MWCO of uncoated Multibore membranes should be at 100,000 Da. Investigating the MWCO value based on the 12 of 15 g to the ncoated 12 of 15 g to the ncoated Membranes 2020, 10, 412 was possible to obta a ufa tu e i fo experimental procedure previously described resulted in an average cut-oﬀvalue of 101,000 Da for the three membranes tested, as shown in Figure 9. Multibore membranes should be at 100,000 Da. Investigating the MWCO value based on the experimental procedure previously described resulted in an average cut-off value of 101,000 Da for the three membranes tested, as shown in Figure 9. Figure 9. Molecular weight cut-off for uncoated PES multibore membranes. 0 10 20 30 40 50 60 70 80 90 100 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 Rejection [%] Molar Mass [kDa] Membrane 01 Membrane 02 Membrane 03 Figure 9. Molecular weight cut-oﬀfor uncoated PES multibore membranes. Figure 9. Molecular weight cut-off for uncoated PES multibore membranes. Figure 9. Molecular weight cut-oﬀfor uncoated PES multibore membranes. The average value is in very good accordance with the manufacturer data. In the single experiments, it became apparent that the individual uncoated membranes diﬀer in a range of 92,000, 96,000, and 116,000 Da. This might be a result of the very small lab modules used in the experiments which only had a surface of <0.01 m2 compared to commercially used membrane modules of several m2. y p y After the coating with 8 DL of PDADMAC/PSS, the mean MWCO of three membranes became 335 Da, with values of 300, 350, and 360 Da, displayed in Figure 10. The pore di 4. Conclusions [24], using the molar mass, resulting in a value of 1.6 nm for the 8 DL-modified membrane. This value indicates a membrane with a pore diameter in the range of a nanofiltration membrane [15]. The permeability data for the modified membranes are in agreement with this calculation, showing results in the range of nanofiltration membranes. However, the rejection of divalent ions (sulfate) is much lower compared to commercial nanofiltration membranes, which have a rejection of >95% [15]. This supports the assumption that the rejection mechanism in LbL membranes is not only dependent on the size exclusion but also on the membrane charge. The results of the zeta potential measurements and the MWCO determination suggest that both the decrease in the pore diameter as well as the change in the surface charge have a major impact on the rejection behaviour. In general, a higher rejection can be associated with a more negative zeta potential, but simultaneously, the membrane has a lower permeability. A less negative zeta potential In this paper, we present the successful adaption and implementation of the zeta potential measurement in multibore membranes. With this innovative method adaption, it was possible to determine the zeta potential in hollow ﬁbre membranes with several capillaries and to show how the zeta potential is altered due to the deposition of PEM. The zeta potential changed depending on the terminating layer of PSS or PDADMAC, leading to a zig-zag pattern. This zig-zag behaviour was also found for the permeability, as well as for the rejection, of a divalent ion (sulfate), showing that a more negative zeta potential can be associated with a higher rejection, while a less negative zeta potential leads to a lower rejection. Additionally, a general trend of an increase in the total values of the zeta potential was observed. This phenomenon could be explained by the more mobile PDADMAC, which has a higher swelling capacity, leading to a migration of the polycation to the surface of the membrane and thus, causing an increased zeta potential. leads to a lower average rejection of divalent ions. Therefore, PSS terminated membranes generally show a better rejection (compared to PDADMAC terminated membranes) due to their negative zeta potential, but at the same time, have a lower permeability. 3.5. Determination of the Molecular Weight Cut-Off 3.5. Determination of the Molecular Weight Cut-Oﬀ This means that the 8 DL coating leads to a reduction in the MWCO of the membrane by more than 99%. These values are consistent with the results of Menne 2017, who also determined the MWCO of modiﬁed hollow ﬁbre PES membranes and measured a mean value of 250 Da for 8 DL coating of PDADMAC/PSS (applied in similar coating conditions, 0.1 M of NaCl) [2]. The pore diameter of the coated membranes was calculated according to Crittenden et al., 2012 [24], using the molar mass, resulting in a value of 1.6 nm for the 8 DL-modiﬁed membrane. This value indicates a membrane with a pore diameter in the range of a nanoﬁltration membrane [15]. The permeability data for the modiﬁed membranes are in agreement with this calculation, showing results in the range of nanoﬁltration membranes. However, the rejection of divalent ions (sulfate) is much lower compared to commercial nanoﬁltration membranes, which have a rejection of >95% [15]. This supports the assumption that the rejection mechanism in LbL membranes is not only dependent on the size exclusion but also on the membrane charge. The results of the zeta potential measurements and the MWCO determination suggest that both the decrease in the pore diameter as well as the change in the surface charge have a major impact on the rejection behaviour. In general, a higher rejection can be associated with a more negative zeta potential, but simultaneously, the membrane has a lower permeability. A less negative zeta potential leads to a lower average rejection of divalent ions. Therefore, PSS terminated membranes generally show a better rejection (compared to PDADMAC terminated membranes) due to their negative zeta potential, but at the same time, have a lower permeability. The increasing zeta potential with the rising number of layers leads to the assumption that the positive charge of the polycation PDADMAC leads to less rejection of sulfate. It is probable that the 70% rejection for 8 DL-modiﬁed membrane is mainly 13 of 15 92,000, riments 13 of 15 92,000, riments Membranes 2020, 10, 412 experiments, it becam 96 000 a d 116 000 D a result of the reduction in pore size to 1.6 nm. This assumption is also supported by further rejection experiments at pH 3. As shown in Figure 4, at pH 3, the IEP is reached, where the net charge of the membrane is zero. 3.5. Determination of the Molecular Weight Cut-Off 3.5. Determination of the Molecular Weight Cut-Oﬀ Filtration of an 8 DL-coated membrane at this pH shows a decline in the sulfate rejection to 30%, compared to 70% at pH 6. Although the rejection of 30% could also probably be reached due to the smaller pore size, it is obvious that the zeta potential has a huge impact on the rejection of the divalent ions, as even a slightly negative zeta potential of −15 mV at pH 6 increases the rejection by 40%. which only had a surface of <0.01 m compared to commercially used membrane modules of several m2. After the coating with 8 DL of PDADMAC/PSS, the mean MWCO of three membranes became 335 Da, with values of 300, 350, and 360 Da, displayed in Figure 10. This means that the 8 DL coating leads to a reduction in the MWCO of the membrane by more than 99%. These values are consistent with the results of Menne 2017, who also determined the MWCO of modified hollow fibre PES membranes and measured a mean value of 250 Da for 8 DL coating of PDADMAC/PSS (applied in similar coating conditions, 0.1 M of NaCl) [2]. Figure 10. Molecular weight cut-off for 8 DL-coated multibore membranes. 0 10 20 30 40 50 60 70 80 90 100 0 500 1000 1500 2000 Rejection [%] Molar Mass [Da] Membrane 01 (8DL) Membrane 02 (8DL) Membrane 03 (8DL) Figure 10. Molecular weight cut-oﬀfor 8 DL-coated multibore membranes. Figure 10. Molecular weight cut-off for 8 DL-coated multibore membranes. Figure 10. Molecular weight cut-oﬀfor 8 DL-coated multibore membranes. rejection experiments at pH 3. As shown in Figure 4, at pH 3, the IEP is reached, where the net charge of the membrane is zero. Filtration of an 8 DL-coated membrane at this pH shows a decline in the sulfate rejection to 30%, compared to 70% at pH 6. Although the rejection of 30% could also probably Author Contributions: Conceptualization: S.D.; methodology: S.D., S.A.K.; experiments: S.A.K., S.D., writing—original draft preparation: S.D.; writing—review and editing: S.D., S.A.K., J.S., M.E. All authors have read and agreed to the published version of the manuscript. Funding: This research received no external funding. References 1. Joseph, N.; Ahmadiannamini, P.; Hoogenboom, R.; Vankelecom, I.F.J. Layer-by-layer preparation of polyelectrolyte multilayer membranes for separation. Polym. Chem. 2014, 5, 1817–1831. [CrossRef] 1. Joseph, N.; Ahmadiannamini, P.; Hoogenboom, R.; Vankelecom, I.F.J. Layer-by-layer preparation of polyelectrolyte multilayer membranes for separation. Polym. Chem. 2014, 5, 1817–1831. [CrossRef] 2. Menne, D. Layer-by-Layer Design of Nanoﬁltration Membranes: Entwicklung von “Layer-by-Layer” Nanoﬁltrationsmembranen. Ph.D. Thesis, RWTH Aachen University, Aachen, Germany, 25 January 2017. 2. Menne, D. Layer-by-Layer Design of Nanoﬁltration Membranes: Entwicklung von “Layer-by-Layer” Nanoﬁltrationsmembranen. Ph.D. Thesis, RWTH Aachen University, Aachen, Germany, 25 January 2017. 3. Blázquez, M.; Tellaetxe, A.; de Grooth, J.; Potreck, J.; van Foeken, C.; Borre, F.H.J.; Gascón, A.J. Life cycle assessment of a layer by layer nanoﬁltration membrane—A case study of the LbLbrane project. In Proceedings of the Avnir conference, Lille, France, 5–6 November 2014. 3. Blázquez, M.; Tellaetxe, A.; de Grooth, J.; Potreck, J.; van Foeken, C.; Borre, F.H.J.; Gascón, A.J. Life cycle assessment of a layer by layer nanoﬁltration membrane—A case study of the LbLbrane project. In Proceedings of the Avnir conference, Lille, France, 5–6 November 2014. 4. Menne, D.; Üzüm, C.; Koppelmann, A.; Wong, J.E.; van Foeken, C.; Borre, F.; Dähne, L.; Laakso, T.; Pihlajamäki, A.; Wessling, M. Regenerable polymer/ceramic hybrid nanoﬁltration membrane based on polyelectrolyte assembly by layer-by-layer technique. J. 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Multilayer Thin Films: Sequential Assembly of Nanocomposite Materials; W Hoboken, NJ, USA, 2012. . Kelly, K.D. Kinetics and Application of Polyelectrolyte Membranes and Multilayers. Ph.D. Th Florida State University, Tallahassee, FL, USA, 10 May 2016. 10. Ghostine, R.A.; Markarian, M.Z.; Schlenoﬀ, J.B. Asymmetric growth in polyelectrolyte multilayers. J. Am. Chem. Soc. 2013, 135, 7636–7646. [CrossRef] [PubMed] 11. The pore di 4. Conclusions The increasing zeta potential with the rising number of layers leads to the assumption that the positive charge of the polycation PDADMAC leads to less rejection of sulfate. It is probable that the 70% rejection for 8 DL-modified membrane is mainly a result of the reduction in pore size to 1.6 nm. This assumption is also supported by further Additional investigations on the MWCO have shown that the pore size of the 8 DL modiﬁed membrane is in the range of a nanoﬁltration. As the permeability data support this determination, the rejection, which was at 70% for an 8 DL coated membrane, did not reach the value of NF membranes, which usually has a sulfate retention >95%. Evidently, the rejection is not only dependent on the smaller pore size of the modiﬁed membrane, but is also strongly inﬂuenced by the less negative zeta potential at 8 DL. eje io e pe i e a p A o i igu e , a p , e IE i ea e , e e e e a ge of the membrane is zero. Filtration of an 8 DL-coated membrane at this pH shows a decline in the sulfate rejection to 30%, compared to 70% at pH 6. Although the rejection of 30% could also probably Author Contributions: Conceptualization: S.D.; methodology: S.D., S.A.K.; experiments: S.A.K., S.D., writing—original draft preparation: S.D.; writing—review and editing: S.D., S.A.K., J.S., M.E. All authors have read and agreed to the published version of the manuscript. 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https://openalex.org/W2136919219	https://epub.ub.uni-muenchen.de/22868/1/1471-2121-15-22.pdf	English	null	Distinct nuclear orientation patterns for mouse chromosome 11 in normal B lymphocytes	BMC cell biology	2,014	cc-by	9,675	Abstract Background: Characterizing the nuclear orientation of chromosomes in the three-dimensional (3D) nucleus by multicolor banding (mBANDing) is a new approach towards understanding nuclear organization of chromosome territories. An mBANDing paint is composed of multiple overlapping subchromosomal probes that represent different regions of a single chromosome. In this study, we used it for the analysis of chromosome orientation in 3D interphase nuclei. We determined whether the nuclear orientation of the two chromosome 11 homologs was random or preferential, and if it was conserved between diploid mouse Pre B lymphocytes of BALB/c origin and primary B lymphocytes of congenic [T38HxBALB/c]N wild-type mice. The chromosome orientation was assessed visually and through a semi-automated quantitative analysis of the radial and angular orientation patterns observed in both B cell types. Results: Our data indicate that there are different preferential patterns of chromosome 11 orientation, which are not significantly different between both mouse cell types (p > 0.05). In the most common case for both cell types, both copies of chromosome 11 were oriented in parallel with the nuclear border. The second most common pattern in both types of B lymphocytes was with one homolog of chromosome 11 positioned with its telomeric end towards the nuclear center and with its centromeric end towards the periphery, while the other chromosome 11 was found parallel with the nuclear border. In addition to these two most common orientations present in approximately 50% of nuclei from each cell type, other orientations were observed at lower frequencies. Conclusions: We conclude that there are probabilistic, non-random orientation patterns for mouse chromosome 11 in the mouse B lymphocytes we investigated (p < 0.0001). Keywords: Chromosome orientation, Three-dimensional nucleus, Nuclear architecture, Fluorescence in situ hybridization, Multicolor banding, Chromosome territory located further towards the center of the nucleus whereas chromosomes with a lower gene-density are concentrated at the nuclear periphery [1,3-6]. Transcription is also thought to play an important role in CT arrangement, with transcriptionally active genes usually located on the edge or outside of CTs and inactive genes found in the interior [7]. Gene expression can also cause chromatin movement in the 3D nucleus, as active genes may loop out of their CT altogether, presumably to access a tran- scription factory [8,9]. The correlation between the radial distribution of CTs and factors such as gene density, replication timing and transcription were examined by Küpper et al. [10]. RESEARCH ARTICLE Open Access * Correspondence: S.Mueller@lrz.uni-muenchen.de; Sabine.Mai@med. umanitoba.ca 2Institute of Human Genetics, University Hospital, Ludwig-Maximilians-University, Goethestr. 29, 80336 Munich, Germany 1Manitoba Institute of Cell Biology, University of Manitoba, Cancer Care Manitoba, 675 McDermot Avenue, Winnipeg, Manitoba, Canada Full list of author information is available at the end of the article © 2014 Schmälter et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Distinct nuclear orientation patterns for mouse chromosome 11 in normal B lymphocytes Ann-Kristin Schmälter1,2, Alexandra Kuzyk1, Christiaan H Righolt1,3, Michaela Neusser2, Ortrud K Steinlein2, Stefan Müller2* and Sabine Mai1* Abstract They found that, in human cell nuclei, gene-density has a dominant impact on the radial distri- bution of CTs. In mouse cell nuclei other factors like Results The mBANDing technique was used to study the nuclear organization of chromosome 11 in a diploid mouse Pre B lymphocyte line of BALB/c origin [21] and in B lympho- cytes of congenic [T38HxBALB/c]N wild-type mice [22]. We visualized mBANDed chromosome 11 in metaphase preparations and chromosome territory (CT) 11 in 3D nu- clei. Over 300 nuclei of both PreB and [T38HxBALB/c]N wild-type mouse lymphocytes were imaged using Axiovi- sion 4.8 software (Carl Zeiss Inc. Canada). After deconvolu- tion using a constrained iterative algorithm [23], all nuclei were analyzed by visual inspection to determine the orien- tation of both chromosome 11 homologs. To validate these results, we performed a semi-quantitative analysis of the ra- dial arrangement of individual mBAND probe distributions on a subset of 45 nuclei per cell type using eADS software [10]. The 3D conformation of 90 individual chromosomes 11 from each of the two cell types was determined by measuring angles between the geometric centers of the dif- ferent mBAND probes in individual chromosome territor- ies. To determine whether cell cycle distribution had an impact on the chromosome 11 orientation patterns seen, cell cycle profiles of both B cell types were measured by FACS analysis. In the present study, we investigated chromosome orien- tation for the first time in the mouse 3D nucleus. We used multicolor banding (mBANDing). A mBAND paint labels regions of a single chromosome with different fluoro- chromes. These different stains ensure that the centromeric end, telomeric end and interstitial regions can be differenti- ated from each other. In the 3D nucleus, the location of each region, and ultimately the orientation of the whole chromosome can, therefore, be determined. mBANDing is commonly used to study intrachromosomal changes in sin- gle chromosomes [17], but can also be applied in studies of nuclear architecture. Using mBAND probes, the degree of condensation of human chromosome 5 was determined in both interphase and metaphase [18], more recently the orientation of human chromosomes in sperm nuclei were analyzed [19]. In the latter study, the radial positions of all 24 CTs and their axial vs. non-axial as well as their linear vs. non-linear, orientations with respect to the sperm tail were determined, as well as the internal organization of chromosome subregions defined by different mBAND probes. A predominantly size-dependent radial arrange- ment was found for entire CTs. Results In addition, in particular for the smaller chromosomes, the authors also reported a gene density correlated orientation. Taken together, their study did not identify a preferential internal orientation of CTs with regard to the telomeric and centromeric end. Background Chromosomes occupy specific regions in the three- dimensional (3D) interphase nucleus, so-called chromo- some territories (CTs) [1]. The radial arrangement of CTs shows cell-type specific differences [1,2]. The arrange- ment of CTs is influenced by many factors, such as chromosome size, gene density and transcription. In lym- phocytes, chromosomes with a high gene-density are Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 2 of 10 between the frequencies of these three patterns of chromo- some 11 orientations in both types of mouse B lymphocytes studied. guanine-cytosine content may, however, play a more im- portant role in determining the radial distribution [11]. The position of each CT is established early in G1 and is maintained throughout interphase with minimal con- strained diffusion [12]. Changes in the position of CTs have, however, been observed during cell differentiation, senescence and tumorigenesis. This occurs for example during adipocyte [13] and human epidermal keratinocyte differentiation [14]. In human fibroblasts, chromosome positions change when a cell becomes quiescent, senes- cent or when it re-enters the cell cycle [15,16]. Chromosome 11 mBANDing in Pre B and T38H wt metaphase preparations Mouse chromosome 11 is approximately 122 Mb in size and has a mean gene density of 18.7 genes/Mb [24]. The mBAND probe set divides chromosome 11 into four over- lapping segments, as illustrated in Figure 1A. The pericen- tromeric region is labeled with Texas Red, the proximal interstitial region in Gold, the distal interstitial region with DEAC and the telomeric region with FITC, respectively. Hybridization of the chromosome 11 mBAND probe to metaphase spreads (Figure 1B) was performed to examine copy number and structural stability of chromosome 11 in both cell types. Twenty metaphases in three independent FISH experiments were analyzed per cell type. We ob- served no structural or numerical aberrations in chromo- some 11 in the cells studied. Figure 1B represents an example of a PreB lymphocyte metaphase, with both copies of chromosome 11 labeled by the mBAND probe. Figure 1C depicts the mBAND profiles of the two chromosomes from a PreB lymphocyte metaphase. All four fluorochromes can be clearly identified. Our aim was to determine and compare the orientation of chromosome 11 in a diploid mouse PreB lymphocyte cell line and in primary B lymphocytes of congenic [T38HxBALB/c]N wild-type mice. Chromosome 11 is a gene dense chromosome [20]. The mBAND paint labels re- gions of chromosome 11 with four different fluorochromes. After fluorescence in situ hybridization (FISH) on 3D pre- served cell nuclei, the location of centromeric, telomeric and interstitial regions and the orientation of chromosome 11 were visually determined for 300 nuclei per cell type. We observed three main patterns of chromosome 11 orien- tations. One arrangement involved chromosome 11 in par- allel with the nuclear border, with neither the telomeric nor centromeric end pointing towards the nuclear center. Alter- natively, the telomeric or the centromeric end of chromo- some 11 were found pointing towards the nuclear center. Our data show that there is no significant difference mBANDing of chromosome 11 in interphase nuclei mBANDing of chromosome 11 in interphase nuclei We performed FISH on 3D preserved cell preparations from the two B cell types. With exception for the distal Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 3 of 10 Schmälter et al. BMC Cell Biology 2014, 15:22 Page 3 of 10 Figure 1 mBAND labeling scheme, metaphase of PreB lymphocytes of BALB/c origin, mBANDed chromosomes 11 and representative [T38HxBALB/c]N wild-type B cell interphase nucleus. A mBAND labeling scheme of mouse chromosome 11. Chromosome 11 is divided into four overlapping segments. Each segment was labeled with a different fluorochrome: the telomeric end in FITC (green), the centromeric end in Texas Red (magenta), and the interstitial segments in DEAC (cyan blue) and Gold (red), respectively. B Metaphase from diploid mouse Pre B lymphocytes of BALB/c origin after hybridization with the chromosome 11 mBAND probe. (Scale bar: 10 μm). C Two chromosome homologs from a different Pre B lymphocyte metaphase showing (from left to right) display colors, false colors and mBAND fluorescence intensity profiles. D Maximum intensity projections of a representative [T38HxBALB/c]N wild-type B cell interphase nucleus, from top to bottom: xy-view (z-projection), 60° and 120° rotation around the y-axis (note: the DEAC labeled probe is not shown because of insufficient signal/noise ratio of the FISH signal. The white line outlines the DAPI stained nucleus). (Scale bars are 5 μm). Figure 1 mBAND labeling scheme, metaphase of PreB lymphocytes of BALB/c origin, mBANDed chromosomes 11 and representative [T38HxBALB/c]N wild-type B cell interphase nucleus. A mBAND labeling scheme of mouse chromosome 11. Chromosome 11 is divided into four overlapping segments. Each segment was labeled with a different fluorochrome: the telomeric end in FITC (green), the centromeric end in Texas Red (magenta), and the interstitial segments in DEAC (cyan blue) and Gold (red), respectively. B Metaphase from diploid mouse Pre B lymphocytes of BALB/c origin after hybridization with the chromosome 11 mBAND probe. (Scale bar: 10 μm). C Two chromosome homologs from a different Pre B lymphocyte metaphase showing (from left to right) display colors, false colors and mBAND fluorescence intensity profiles. D Maximum intensity projections of a representative [T38HxBALB/c]N wild-type B cell interphase nucleus, from top to bottom: xy-view (z-projection), 60° and 120° rotation around the y-axis (note: the DEAC labeled probe is not shown because of insufficient signal/noise ratio of the FISH signal. The white line outlines the DAPI stained nucleus). (Scale bars are 5 μm). (Table 1). mBANDing of chromosome 11 in interphase nuclei There was no significant difference in the occur- rence of this orientation pattern between both lymphocyte types against all other patterns combined (p = 0.20). The second most common orientation observed was with one homolog of chromosome 11 orientated with its telomeric end pointing towards the nuclear center, while the other chromosome 11 was parallel with the nuclear border (“PC”) (20.5% and 26.1% in PreB and [T38HxBALB/c]N wild-type mouse lymphocytes, respectively). There was no significant difference in the frequency of this orientation pattern seen between both types of lymphocytes (p = 0.13) (Table 1). One chromosome 11 pointing with its centro- meric end to the center and the other in parallel with the nuclear border (“PT”) was the third most common orienta- tion we observed. The occurrence of this orientation pattern did not differ significantly in both types of lympho- cytes (p = 0.05) with a frequency of 16.3% of the PreB nuclei and of 10.4% of the [T38HxBALB/c]N wild-type lympho- cyte nuclei (Table 1). In 10.7% of PreB and 5.8% of [T38HxBALB/c]N wild-type lymphocyte nuclei, both cop- ies of chromosome 11 were orientated with their centro- meric ends to the nuclear center (“TT”) (Table 1). Furthermore, in 9.1% in PreB and 7.3% [T38HxBALB/c]N wild-type lymphocyte nuclei one chromosome 11 pointed towards the nuclear center with its telomeric end and the other points towards the center with its centromeric end (“CT”) (Table 1). The scenario that both chromosomes 11 were orientated with their telomeric ends to the nuclear center (“CC”) was observed in 6.1% of the PreB and 18.5% of the [T38HxBALB/c]N wild-type mouse lymphocyte nu- clei (Table 1). interstitial probe labeled with DEAC, all mBAND probes consistently showed the identification of specific FISH signals that were clearly distinguishable from non- specific background. We therefore excluded the DEAC labeled mBAND probe from all further measurements. Image z-stacks from 307 nuclei of PreB and 303 nuclei of [T38HxBALB/c]N wild-type mouse lymphocytes were captured and deconvolved (Materials and Methods). Figure 1D shows a representative chromosome 11 mBANDing image of a [T38HxBALB/c]N wild-type B lymphocyte 3D nucleus. 3D reconstructed images and movies representing various orientation patterns can be viewed in Additional files 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12. Orientation of chromosome 11 in the 3D nucleus as examined by visual inspection The nucleus is illustrated in blue; the black circles divide the nucleus into central, intermediate and peripheral regions (PP = both homologs parallel to the nuclear periphery, PC/PT = one homolog is located in parallel to the periphery, while the other is oriented with its centromeric (red)/telomeric (green) end towards the nuclear periphery, TT = both homologs point with their telomeric (green) ends towards the periphery, CT = one homolog with its centromeric (red) end and the other homolog with its telomeric (green) end pointing towards the nuclear periphery, CC = both homologs point with their centromeric (red) ends towards the nuclear periphery, ). PP, PC and PT in the first row of Figure 2 are observed most frequently. to the nuclear border (“PP”); (2) one copy of chromo- some 11 in parallel to the border and the other copy pointing with its telomeric end towards the nuclear cen- ter (“PC”); (3) one copy of chromosome 11 in parallel to the border and the other copy pointing with its centro- meric end towards the center (“PT”). Table 1 shows the frequencies of the different orientation patterns in the two cell types and 2x2 chi square p-values demonstrat- ing the significant difference of each pattern with respect to all other orientation patterns combined. Orientation of chromosome 11 in the 3D nucleus as examined by visual inspection Using mBANDing, we were able to analyze the chromo- some orientation of mouse chromosome 11 subregions for the first time in 3D interphase nuclei. We deter- mined the frequency of nuclear chromosome orientation patterns by visual inspection in all captured nuclei from PreB lymphocytes and [T38HxBALB/c]N wild-type mice. mBAND territories were designated as parallel (“P”) to the nuclear surface when no chromosome end was pointing towards the nuclear center or the periph- ery. A homolog of chromosome 11 was classified as “C” when the centromeric region was localized closest to the nuclear border and “T when its telomeric end showed the most peripheral positioning. Figure 2 provides a car- toon illustration for the various orientation patterns ob- served. The most frequent pattern observed was with both copies of chromosome 11 located in parallel with the nuclear border (“PP”) (37.3% and 31.9%, respectively) By visual inspection we observed three main patterns of chromosome 11 orientation in the 3D interphase nu- cleus: (1) both homologs of chromosome 11 in parallel Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 4 of 10 Figure 2 Cartoon illustration of different possibilities of chromosome 11 orientation pairs in the nucleus. The representative chromosome is composed of four differently colored segments; green segments represent telomeric ends, magenta segments centromeric ends, proximal and distal (not measured) interstitial segments are shown in cyan blue and red, respectively. The nucleus is illustrated in blue; the black circles divide the nucleus into central, intermediate and peripheral regions (PP = both homologs parallel to the nuclear periphery, PC/PT = one homolog is located in parallel to the periphery, while the other is oriented with its centromeric (red)/telomeric (green) end towards the nuclear periphery, TT = both homologs point with their telomeric (green) ends towards the periphery, CT = one homolog with its centromeric (red) end and the other homolog with its telomeric (green) end pointing towards the nuclear periphery, CC = both homologs point with their centromeric (red) ends towards the nuclear periphery, ). PP, PC and PT in the first row of Figure 2 are observed most frequently. Figure 2 Cartoon illustration of different possibilities of chromosome 11 orientation pairs in the nucleus. The representative chromosome is composed of four differently colored segments; green segments represent telomeric ends, magenta segments centromeric ends, proximal and distal (not measured) interstitial segments are shown in cyan blue and red, respectively. Orientation of chromosome 11 in the 3D nucleus as examined by visual inspection There was no significant difference in the occurrence of the orientation patterns observed between both B cell types (p > 0.05), with the exception of two minor patterns of orienta- tion: when both copies of chromosome 11 pointed with Table 1 Frequencies of orientation patterns of chromosome 11 PreB [%] [T38Hx BALB/c]N wt [%] Chi-Square [p-value] Both homologs in parallel to the nuclear border (PP) 37.3 31.9 0.20 One homolog points with its centromere to the nuclear periphery, the other is parallel to the nuclear border (PC) 20.5 26.1 0.13 One copy points with its telomere to the nuclear periphery, the other is parallel to the nuclear border (PT) 16.3 10.4 0.05 Both homologs point with their telomeric end to the nuclear periphery (TT) 10.7 5.8 0.04 One copy points with its telomeric end, and the other copy with centromeric end, to nuclear periphery (CT) 9.1 7.3 0.45 Both copies point with their centromeric ends to the nuclear periphery (CC) 6.1 18.5 <0.0001 Orientation patterns of chromosome 11 and the frequency of orientation in Pre B lymphocytes of BALB/c origin and in B cells of congenic [T38HxBALB/c]N wild-type mice analyzed by visual inspection. A 2x2 Chi-Square test with p-value of >0.05 indicates that the frequency of the orientation is not significantly different between the two cell types. Each pattern was tested between the two cell types against all other patterns combined and the expected values were based on the marginal totals. There is a significant difference in the percentage of cells that display these major 2x3 orientation patterns between the two cell types (p <0.0001). (P = parallel, C = centromere points to periphery, T = telomere points to periphery). Table 1 Frequencies of orientation patterns of chromosome 11 Table 1 Frequencies of orientation patterns of chromosome 11 Orientation patterns of chromosome 11 and the frequency of orientation in Pre B lymphocytes of BALB/c origin and in B cells of congenic [T38HxBALB/c]N wild-type mice analyzed by visual inspection. A 2x2 Chi-Square test with p-value of >0.05 indicates that the frequency of the orientation is not significantly different between the two cell types. Each pattern was tested between the two cell types against all other patterns combined and the expected values were based on the marginal totals. There is a significant difference in the percentage of cells that display these major 2x3 orientation patterns between the two cell types (p <0.0001). Orientation of chromosome 11 in the 3D nucleus as examined by visual inspection (P = parallel, C = centromere points to periphery, T = telomere points to periphery). Orientation patterns of chromosome 11 and the frequency of orientation in Pre B lymphocytes of BALB/c origin and in B cells of congenic [T38HxBALB/c]N wild-type mice analyzed by visual inspection. A 2x2 Chi-Square test with p-value of >0.05 indicates that the frequency of the orientation is not significantly different between the two cell types. Each pattern was tested between the two cell types against all other patterns combined and the expected values were based on the marginal totals. There is a significant difference in the percentage of cells that display these major 2x3 orientation patterns between the two cell types (p <0.0001). (P = parallel, C = centromere points to periphery, T = telomere points to periphery). Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 5 of 10 their centromeric (“TT”) (p = 0.04) or telomeric (“CC”) (p < 0.0001) ends towards the nuclear center (Table 1). A 2x3 contingency analysis of just the three major orientation patterns indicated that there was a signifi- cant difference in the distribution of the three patterns between the two cell type (p < 0.001). PreB and [T38HxBALB/c]N wild-type chromosomes using DistAngle software allowed us to determine whether the chromosome territory is preferentially straight or angulated during interphase. Mean measured angles in PreB cells (107°) were very similar compared to [T38HxBALB/c]N wildtype B cells (106°) and showed no statistically signifi- cant differences (p = 0.99). In both cell types we observed a broad distribution of measured angles (Figure 3A and 3B). 19% of PreB and 15% of [T38HxBALB/c]N wild-type B cell CT11 showed angles between 0 and 60° and were con- sidered as rather angulated. 47% of PreB and 43% of [T38HxBALB/c]N wild-type B cell CT11 showed angles be- tween 121° and 180° and indicate a trend towards a straight configuration (Figure 3A and B). their centromeric (“TT”) (p = 0.04) or telomeric (“CC”) (p < 0.0001) ends towards the nuclear center (Table 1). A 2x3 contingency analysis of just the three major orientation patterns indicated that there was a signifi- cant difference in the distribution of the three patterns between the two cell type (p < 0.001). Cell cycle profiles When comparing the individual results of the quanti- tative analysis to the analysis by visual inspection of all 45 image per cell type we found, however, that only 30/ 45 (67%) nuclei of [T38HxBALB/c]N wild-type mouse lymphocyte and 28/45 (62%) nuclei of Pre B lympho- cytes showed concordant results with both methods. To investigate whether cell cycle stages impact on the nuclear orientation and orientation patterns of mouse chromosome 11, we carried out fluorescent activated cell sorter (FACS) analysis of cell cycle profiles. The DNA pro- file of the PreB lymphocytes of BALB/c origin is shown in Figure 4A and the DNA profile of the [T38HxBALB/c]N wild-type mouse B cells is shown in Figure 4B. As the PreB lymphocytes were kept proliferating in culture, the cells are distributed throughout the different cell cycle phases with 45.08% in G0/G1, 44.18% in S and 21.80% in G2/M (Figure 4A). Contrastingly, the vast majority of the primary B cells are found in G0/G1 (94.76%) with only 5.22% in S and 2.38% in G2/M (Figure 4B). Semi-automated quantitative analysis of mBAND 11 orientation in mouse lymphocytes To validate the above results, we analyzed nuclear orien- tation patterns of individual chromosomes 11 for a sub- set of 45 nuclei per cell type with the semi-automated software package eADS [9]. The mean radial arrange- ment of telomeric, interstitial and centromeric mBAND regions was determined in these 45 nuclei from both B cell types. We measured the 3D FISH signal distance to the nuclear surface in nm using eADS software [10], and then transformed these values to relative values (%) by normalizing it to the nuclear radius. Additional files 13, 14, 15 and Additional file 16: Table S1 show the results of the semi-automated software assisted analysis: The data indicate that this semi-quantitative analysis identi- fies, similar to the visual inspection, three most frequent orientation patterns for mouse chromosome 11. We also calculated the angle difference between hom- ologous CT11 in individual nuclei. Again, we observed a broad distribution of measured angle differences in both B cell types ranging from 4°-136° (Figure 3C). Mean angle differences between homologs in PreB cells (51°) were slightly larger compared to [T38HxBALB/c]N wild- type B cells (43°), but no statistically significant differ- ences were observed (p = 0.88). 3D conformation of mouse chromosome 11 on the identification and frequency of three different orientation patterns but did not coincide for each nu- cleus measured. This result points to differences in the interpretation of mBAND data by the user (visual inspection) and by the user performing semi-automated analysis that in- cludes the manual segmentation of the regions of inter- est. During this procedure, some pixels might have been discarded or identified as artifacts when cutting out the bands separately in different channels with no overview of the whole chromosome. It was not possible to distin- guish low grey levels from each other or background pixels with value 0, which may lead to mistakes in the manual segmentation. The pixels of the bands were measured with respect to the nucleus stained in DAPI. If DAPI staining was weaker in some cells than in others, then the subsequent identification of the positions of the band pixels are measured inaccurately. Due to the lack of a fully automated program, these current measurements are only indicative of varied chromosome orientation pat- terns in the cells examined, but no absolute measure of chromosome band positioning. We are currently working towards a fully automated program. Figure 4 Cell cycle profiles determined by FACS analysis. The graphs illustrate the cell count in the different phases of the cell cycle, which correlates to the propidium iodide intensity. A The cell cycle profile of PreB lymphocytes of BALB/c origin. 45.08% of cells are found in G0/G1, 44.18% in S and 21.80% in G2/M. B The cell cycle profile of primary B cells from [T38HxBALB/c] wild-type mice. 94.76% of cells are found in G0/G1, 5.22% in s and 2.38% in G2/M. The PreB cells were kept proliferating in culture, while the majority (94.76%) of the primary B cells were directly isolated from the mice (kept under specific pathogen free (SPF) conditions) and are in G0/G1 of the cell cycle; this was confirmed through FACS cell cycle analysis (Figure 4). In cycling PreB cells no correlation was found between cell cycle stage and radial orientation of mBAND CT. We have not determined the cell cycle stage of the in- dividual nuclei, e.g. 3D conformation of mouse chromosome 11 We evaluated the 3D conformation of chromosome 11 in the two lymphocyte cell types using the software eADS [9]. We measured the angles between the geometric centers of centromeric, interstitial and telomeric mBAND territories using the interstitial segment as apex. Analysis of each 90 Figure 3 3D conformation analysis of chromosome 11 mBAND territories in Pre B lymphocyte nuclei and in B cell nuclei of congenic [T38HxBALB/c]N wild-type mice using measured angles between geometric centers of centromeric, interstitial (apex) and telomeric chromosome band territories in individual chromosomes A Distribution of measured angles in each 90 PreB and [T38HxBALB/c]N wild-type B individual chromosomes 11. B Bar diagram showing the percentage of angulated chromosomes with telomeric band territories in close proximity to centromeric regions (0-60° = chromosome 11 territory angulated), intermediate (61-120°) and straight chromosomes (121-180° = chromosome 11 territory straight) in the two cell lines. C Distribution of calculated angle differences between homologous CT pairs as a measure for the similarity of the 3D conformation of the two chromosomes 11 CT in individual nuclei. Figure 3 3D conformation analysis of chromosome 11 mBAND territories in Pre B lymphocyte nuclei and in B cell nuclei of congenic [T38HxBALB/c]N wild-type mice using measured angles between geometric centers of centromeric, interstitial (apex) and telomeric chromosome band territories in individual chromosomes A Distribution of measured angles in each 90 PreB and [T38HxBALB/c]N wild-type B individual chromosomes 11. B Bar diagram showing the percentage of angulated chromosomes with telomeric band territories in close proximity to centromeric regions (0-60° = chromosome 11 territory angulated), intermediate (61-120°) and straight chromosomes (121-180° = chromosome 11 territory straight) in the two cell lines. C Distribution of calculated angle differences between homologous CT pairs as a measure for the similarity of the 3D conformation of the two chromosomes 11 CT in individual nuclei. Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 6 of 10 Figure 4 Cell cycle profiles determined by FACS analysis. The graphs illustrate the cell count in the different phases of the cell cycle, which correlates to the propidium iodide intensity. A The cell cycle profile of PreB lymphocytes of BALB/c origin. 45.08% of cells are found in G0/G1, 44.18% in S and 21.80% in G2/M. B The cell cycle profile of primary B cells from [T38HxBALB/c] wild-type mice. 94.76% of cells are found in G0/G1, 5.22% in s and 2.38% in G2/M. 3D conformation of mouse chromosome 11 by BrdU, so we can only state “no differ- ence between cycling and quiescent cells” The influences of differentiation and gene density on the nuclear architecture and on the radial orientation of gene dense telomeric chro- matin appear to be a more likely cause for the observed dif- ferences between the two cell types. Gene density can be ruled out as cause for differences because it is the same in both cell types. This leaves differentiation as cause, which is linked to transcription. The comparison of the distinct cell cycle profiles ob- served for the [T38HxBALB/c]N wild-type mouse B cells and PreB cells indicates that the chromosome 11 orientation is independent of cell cycle profiles in the two B cell types examined in our study. The comparison of the distinct cell cycle profiles ob- served for the [T38HxBALB/c]N wild-type mouse B cells and PreB cells indicates that the chromosome 11 orientation is independent of cell cycle profiles in the two B cell types examined in our study. Discussion We used mBANDing for the first time to study chromo- some orientation in the mouse 3D B cell nucleus. This technique has mostly been used previously to detect intrachromosomal rearrangements such as inversions, translocations and deletions in metaphase spreads [17]. One previous study used mBANDing to examine the spatial arrangements and the 3D configuration of CT in human sperm nuclei, demonstrating that mBANDing is an excellent technique to determine the 3D nuclear orientation of chromosomes [19]. Two alternative ap- proaches were recently described, which could also be ap- plied for a 3D topology analysis of mouse chromosomes, or could even be combined with the mouse mBAND probe set to further enhance the subregional resolution of the FISH banding probe set. One first approach used evo- lutionary rearranged chromosomes as subchromosomal probes for 3D-FISH [25], the second pooled BAC probes for predefined genomic regions or interest [26]. Transcription may influence chromosome position. Regions of chromatin have been shown to change pos- ition in the nucleus with activation and increased levels of transcription [8,27,28]. Transcription factories can also alter the arrangement of chromatin as genes may loop out of their territory to be transcribed [29] or mul- tiple genes will meet to be co-transcribed in a single transcription factory [30]. The access to transcription factories , therefore, may be relevant for the orientation of chromosomes in the nucleus. A possible rotation of a chromosome in the nucleus may be an alternate way in which active regions can access the transcription ma- chinery. For example, chromosome rotation could occur around the centromeric or telomeric end. This would Using a mouse mBAND 11 probe set, we first deter- mined chromosome 11 orientation by visual inspection. We then conducted a semi-automated quantitative ana- lysis using the software eADS [9]. Both methods agreed Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 7 of 10 most frequent pattern was with both homologs posi- tioned in parallel to the nuclear border in both types of B lymphocytes. Alternatively the telomeric or centro- meric end was found pointing towards the nuclear periphery or center. Overall, chromosome orientation appears to be a non-random feature of the genome in interphase nuclei. allow genes that lay close to rotation axis to remain in a stable position whereas those further away will undergo a positional change. Discussion Alternatively, or in addition to rota- tion, a chromosome territory may adopt different large- scale folding patterns in order to reach this goal, which may result in straight or angulated 3D conformations. Additionally, the active state of chromatin with regards to methylation, acetylation and other post-translational histone modifications, may not only alter chromatin con- densation but also orientation. To test for the effects of transcription on chromosome orientation was beyond the scope of this study. 2D and 3D fixation 2D chromosome fixation was conducted as described by Mai and Wiener [32]. Primary B cells and PreB lympho- cytes were centrifuged for 10 minutes at 1000 rpm. The pellet was resuspended in 0.075 M KCl for 30 minutes. Next the cells underwent centrifugation for 10 minutes at 1000 rpm and then a drop fixation with 3:1 methanol to acetic acid. After resuspension of the pellet, cells were dropped onto slides. In this study we examined, for the first time, chromo- some orientation in the murine 3D nucleus using mBAND- ing. We found that the orientation of chromosome 11 in PreB lymphocytes of BALB/c origin and primary B cells of congenic [T38HxBALB/c]N wild-type mice varies between distinct patterns of orientation. Gene density has been thought to be the primary factor influencing lymphocyte CT arrangement. We find, however, that it is not sufficient to determine the orientation of mouse chromosome 11 in B cells. Because gene-density is stable, it cannot account for differences in the three major patterns of chromosome 11 orientation we observed. Although we did not find orienta- tion pattern and gene-density to be linked for chromosome 11 in both B-cell models, this may not hold true for all mouse chromosomes and cell types and should be assessed for other chromosomes and non B lineage cell types. At present, our findings allow us to conclude that chromo- somes may display distinct orientations in the interphase nucleus of diploid mouse B cells during some stages of dif- ferentiation, but the underlying mechanisms require further investigation. 3D nuclei fixation was conducted as described by Solovei et al. [33]. Primary B cells and PreB lymphocytes were centrifuged for 10 minutes at 1000 rpm. After re- suspension of the pellet, cells were applied to slides. One hour later, the slides were washed in 1 × phosphate- buffered saline (PBS), 0.3xPBS and then incubated in freshly prepared 3.7% formaldehyde. Next the slides were washed in 0.05% Triton-X-100/1xPBS, followed by a wash in 0.5% Triton-X-100/1xPBS. The slides were then incubated in 20% Glycerol/1×PBS, for at least an hour, and subsequently underwent repeated freeze/thaw cycles in liquid nitrogen. Afterwards, the slides were washed in 0.05% Triton-X-100/1xPBS, followed by incu- bation in 0.1 M HCl. After washing the slides in 2× sa- line sodium citrate buffer (SSC) they were kept at least one hour in 50% formamide/2×SSC at 4°C. Cell harvest and cell culture Mouse PreB lymphocytes of BALB/c origin [21] were cultured in RPMI 1640 media supplemented with 10% heat-inactivated fetal bovine serum (FBS), 1% penicillin- streptomycin, 1% L-Glutamine, 1% sodium pyruvate and 0.1% β-mercaptoethanol (Invitrogen/Gibco, Burlington, ON, Canada). Cells were incubated at 37°C. In approximately 45% of the nuclei examined, it was observed that the two homologs of chromosome 11 did not have the same orientation. This may be an innate protection mechanism of the cell. It has been shown by Heride et al. [31] that chromosomes are non-randomly closer to a heterologue than a homologue. This led them to propose that it may be evolutionarily important to position chromosomes in this way to avoid homologous recombination and to avoid damaging both copies of a chromosome by a single genotoxic stress event. The same argument may explain why two copies of a chromosome may adopt different orientations in the nucleus. Primary B cells were harvested from spleens of three 6 to 8 weeks old congenic [T38HxBALB/c]N wild-type mice [22]. Procedures were conducted according to Animal Protocol 11-019, approved by Central Animal Care Ser- vices, University of Manitoba (Winnipeg, MB, Canada). Multicolor banding h The mBANDing probe for mouse chromosome 11 was previously described by Benedek et al. [35] and was pur- chased from Metasystems (Altussheim, Germany) for the present experiments. The pericentromeric region is labeled with Texas Red, the proximal interstitial region in Gold, the distal interstitial region in DEAC and the telomeric region with FITC, respectively. First, slides were equilibrated in 2×SSC, followed by an RNAase A treatment (100 μg/ml) in 2×SSC for one hour at 37°C, and then incubation in freshly prepared 0.01 M HCl with 100 μg/ml pepsin for two minutes. After washing the slides in 1xPBS, they were pretreated in 1% formal- dehyde in 1×PBS/50 mM MgCl2, followed by a wash in 1×PBS. For denaturation, the slides were incubated in 0.1×SSC, and then transferred into 2×SSC at 70°C for 30 minutes. After the solution was cooled down to 37°C, the slides were transferred to 0.1×SSC and then dena- tured in 0.07 M NaOH at room temperature for one mi- nute. Before dehydration in ethanol (30%, 50%, 70% and 90%), the slides were placed in 0.1×SSC and then 2×SSC at 4°C. Next, the mBANDing probe was applied as recommended, sealed to the slide with rubber cement and incubated for two days at 37°C. Post-hybridization washes included 1×SSC at 75°C and in 4×SSC/0.05% Tween20. The chromatin was counterstained with 4’6’-dia- midino-2-phenylindole (DAPI) and mounted with ProLong Gold antifade (Invitrogen/Gibco, Burlington, ON, Canada). For the 3D image acquisition, an AxioImager Z2 micro- scope (Carl Zeiss Inc. Canada) equipped with the same fil- ter sets as for 2D image acquisition and an AxioCam MRm (Carl Zeiss Inc. Canada) was used, combined with Axiovi- sion 4.8 software (Carl Zeiss Inc. Canada). To reconstruct a 3D image, z-stacks of 80 slices, with 200 nm axial distance and 102 nm x/y pixel size were acquired. Deconvolution was performed with a constrained iterative algorithm [23] using Axiovision 4.8 software (Carl Zeiss Inc. Canada). The chromosome orientation was determined by visual inspec- tion based on the mBAND FISH pattern. Quantitative semi-automated analysis Quantitative measurements of mean radial mBAND probe distributions with respect to the nuclear border were performed using the software eADS, a 3D distance measurement tool described in detail by Küpper et al. [10]. In short, the pixels are manually classified as being band or not and being nucleus or not. Multicolor banding h The Euclidean distance between the nuclear surface and each bands pixel is then measured from these images. For the probe distributions each pixel in each band is used, for the de- termination of the orientation only the smallest distance for each band is used. The orientation was determined by the difference in radial position of the telomeric and centromeric band. When the difference was smaller than N percentpoints it was called parallel. The software DistAngle [36] was employed to measure 3D angles between geomet- ric centers of different mBAND probes from individual chromosomes. Fluorescence-activated cell sorting (FACS) analysis We analyzed for the first time the orientation of mouse chromosome 11 with respect to the telomeric and centro- meric ends of chromosomes in the 3D interphase nucleus. Distinct orientation patterns of CT 11 in PreB lympho- cytes of BALB/c origin and primary B cells of congenic [T38HxBALB/c]N wild-type mice were observed. The Fluorescence-activated cell sorting (FACS) analysis FACS analysis was conducted as described by Caporali et al. [34]. Briefly, PreB lymphocytes BALB/c origin and primary B cells of [T38HxBALB/c]N wild-type mice were fixed in 70% cold ethanol and incubated at 4°C overnight. The following day, the pellet was washed Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Page 8 of 10 436 nm/20 nm, Emission CWL/Bandwidth: 480 nm/ 30 nm, Chroma Series No.: 31000 Series), the region pseudo-colored in red by a 11006v3 Gold filter (Excita- tion CWL/Bandwidth: 350 nm/50 nm, Emission CWL/ Bandwidth: 515 nm/nm, Chroma Series No.: 11000 Series) and the region pseudo-colored in magenta by a 41004 Texas Red® filter (Excitation CWL/Bandwidth: 560 nm/55 nm, Emission CWL/Bandwidth: 645 nm/ 75 nm, Chroma Series No.: 41000 Series). twice with 1% FBS in 1xPBS after centrifugation at 1200 rpm for ten minutes. The final pellet was resus- pended in 1×PBS and stained with propidium iodide (1 μg/mL) (Sigma Aldrich, Oakville, ON, Canada). Flow cytometry was used to analyze the cell cycle profiles using a FACSCalibur (Becton Dickinson, Mississauga, ON, Canada). Image acquisition Two-dimensional image acquisition was performed using an Axioplan 2 microscope (Carl Zeiss Ltd., Toronto, ON, Canada) with a 63x/1.4 oil objective lens (Carl Zeiss Ltd., Toronto, ON, Canada) and the ISIS-FISH imaging system 5.0 SR 3 (Metasystems Group Inc. Boston, MA, USA). A DAPI filter was used to visualize the chromosomal counter- stain. To detect the four regions of chromosome 11 that were labeled with different fluorochromes (DEAC, FITC, Gold and TexasRed, respectively), appropriate narrow band pass filters were used (Chroma Technologies). The region pseudo-colored in green was detected by a SP-101 fluores- cein isothiocyanate (FITC) filter (Excitation CWL/Band- width: 471 nm/39 nm, Emission CWL/Bandwidth: 522 nm/ 40 nm, Chroma Series No.: SP100), the region pseudo- colored in cyan by a 31036v2 7-diethylaminocoumarin-3- carboxylic acid (DEAC) filter (Excitation CWL/Bandwidth: Statistical analysis h d ff The different orientations of chromosome 11 were com- pared to each other and compared between the different types of lymphocytes using all of the following tests: Chi-Square, Likelihood Ratio Chi-Square, Continuity Adj Chi-Square and Mantel Haenszel Chi-Square analysis. The p-values of test results shown here for the compari- son of different orientations are the Chi-Square values. All other tests yielded similar results (data not shown). The Mann-Whitney rank sum test was used to deter- mine statistically significant differences in the median ra- dial arrangement of mBAND FISH signals. A p-value <0.05 was considered significant. Page 9 of 10 Page 9 of 10 Schmälter et al. BMC Cell Biology 2014, 15:22 http://www.biomedcentral.com/1471-2121/15/22 Author’s contribution B In T38wt cells mean absolute and relative probe distances to nuclear surface were 1906 nm (56%) for the telomeric region, 1617 nm (47%) for the interstitial and 1522 nm (44%) for the centromeric region. A small percentage of nuclei shows band signals beyond the nuclear border. This is probably due to weak nuclear DAPI stain measured by the software eADS (see Discussion). Additional files nucleus. We chose a 15% cut-off level because this is approximately twice the distance between consecutive image z-sections of 200 nm. Consequently, in chromosome 11 territories where at least one of the measured relative distances between mBAND territories would exceed 15% the CT was assigned an orientation with either the telomeric or the interstitial or the centromeric end pointing towards the nuclear periphery or center. (P = parallel, C = centro- mere points to periphery, I = interstitial is most peripheric, T = telomere points to periphery). Additional file 1: 3D movie of a [T38HxBALB/c]N wild type B cell nucleus with the orientation pattern “CP”. Additional file 2: 3D movie illustrating a Pre B nucleus with the orientation pattern “PP”. Additional file 3: 3D view of a Pre B nucleus with the orientation pattern “CT”. One chromosome 11 is oriented with its centromeric end (red) towards the nuclear center, whereas the other chromosome 11 is oriented with its telomeric end (green) towards the center. Additional file 15: Relative radial orientation of 45 homologous chromosome 11 mBAND CT pairs from each 45 PreB and [T38HxBALB/c]N wildtype B cell interphase nuclei. Frequencies (%) of homologous CT pairs showing different combinations of radial orientations. (P = parallel, C = centromere points to periphery, I = interstitial is most peripheric, T = telomere points to periphery). Additional file 4: 3D movie illustrating a Pre B nucleus with the orientation pattern “TP”. Additional file 5: 3D view of a Pre B nucleus with the orientation pattern “PP”. Both copies of chromosome 11 are located in parallel to the nuclear periphery. Additional file 16: Table S1. Results of semi-automated quantitative analysis using the software eADS. Frequencies of relative orientation patterns of homologous chromosomes in individual nuclei. (P=parallel, C=centromere points to periphery, I=interstitial is most peripheric, T=telomere points to periphery). Additional file 6: 3D movie illustrating a Pre B nucleus with the orientation pattern “TT”. Additional file 7: 3D view of a Pre B nucleus with the orientation pattern “TP”. One chromosome 11 is oriented with its centromeric end (red) towards the nuclear center and with its telomeric end (green) towards the periphery, the other chromosome is located in parallel to the nuclear periphery. Acknowledgements h d This study was supported by CIHR. AKS received a travel award from Bayer. We thank Mary Cheang for statistical analysis and Dr. Ludger Klewes for his help with FACS analysis. Competing interests Th h d l h Competing interests The authors declare that they have no competing interests. The authors declare that they have no competing interests. Additional file 10: 3D view of a [T38HxBALB/c]N wild type B cell nucleus with the orientation pattern “CP”. One chromosome 11 is located in parallel to the nuclear periphery, the other is oriented with its telomeric end (green) towards the center. Author details 1 1Manitoba Institute of Cell Biology, University of Manitoba, Cancer Care Manitoba, 675 McDermot Avenue, Winnipeg, Manitoba, Canada. 2Institute of Human Genetics, University Hospital, Ludwig-Maximilians-University, Goethestr. 29, 80336 Munich, Germany. 3Department of Imaging Physics, Delft University of Technology, Delft, The Netherlands. Received: 4 September 2013 Accepted: 30 May 2014 Published: 12 June 2014 Received: 4 September 2013 Accepted: 30 May 2014 Published: 12 June 2014 Additional file 14: Normalized (% distance to nuclear surface) radial centromeric, interstitial and telomeric mBAND probe distributions in individual chromosomes from each 45 A PreB and B [T38HxBALB/c]N wild-type B cell nuclei using eADS software. The 3D FISH signal distance to the nuclear surface in nm was transformed to relative values (%) by normalization using the nuclear radius as reference. Each colored dot represents the radial position of the geometric center from an individual chromosome 11. The two chromosome 11 homologs from each nucleus are shown side by side. Nuclei with a similar radial orientation of the two homologs are depicted in clusters separated by bold vertical lines. mBAND territories were designated as parallel (“P”) to the nuclear surface when each of the measured relative radial distances between centromeric, telomeric and interstitial chromosome segments was less than 15% of the nuclear radius. This equals to approximately a 400-500 nm radial distance depending on the size of the respective Abbreviations d 2D: Two-dimensional; 3D: Three-dimensional; CT: Chromosome territory; DAPI: 4’6’-diamidino-2-phenylindole; DEAC: 7-diethylaminocoumarin-3- carboxylic acid; FACS: Fluorescence-activated cell sorting; FISH: Fluorescent in situ hybridization; FITC: Fluorescein isothiocyanate; FBS: Fetal bovine serum; mBANDing: Multicolor banding; PBS: Phosphate-buffered saline; SSC: Saline sodium citrate buffer. Additional file 8: 3D movie illustrating a Pre B nucleus with the orientation pattern “CC”. Additional file 9: 3D view of a Pre B nucleus with the orientation pattern “TT”. Both copies of chromosome 11 are pointing with their centromeric ends (red) towards the nuclear center, whereas their telomeric ends (green) are pointing towards the periphery. Author’s contribution AKS carried out 2D and 3D cell preparation and fixation, mBANDing, image acquisition, visual inspection, conducted the analysis by the software eADs and drafted the manuscript. AK contributed to FACS, contributed to 2D and 3D fixation of cells and to the writing of the manuscript. CR contributed to the discussion, created 3D movies, assisted in data analysis and contributed to writing the manuscript. MN participated in the analysis by eADS. OS is AKS’ co-supervisor. SMü conducted the analysis by eADS and contributed to writing the paper. SMa designed the study and helped to draft the manuscript. All authors read and approved the final manuscript. Additional file 11: 3D movie of a [T38HxBALB/c]N wild type B cell nucleus with the orientation pattern “CP”. Additional file 12: 3D view of a [T38HxBALB/c]N wild type B cell nucleus with the orientation pattern “CC”. Both copies of chromosome 11 are pointing with their telomeric ends (green) towards the nuclear center. Additional file 13: Mean radial distribution of the centromeric, interstitial and telomeric mBAND FISH signals in A PreB and B T38wt cell nuclei. The 3D FISH signal distance to the nuclear surface was measured nm using eADS software (Küpper et al., 2007) (n = number of nuclei, nm = nanometer). A In PreB cells the mean radial position of the telomeric segment was at 2016 nm and 42% relative distance from the nuclear surface, the interstitial segment at 2178 nm (46%) and the centromeric segment at 2151 nm (45%). B In T38wt cells mean absolute and relative probe distances to nuclear surface were 1906 nm (56%) for the telomeric region, 1617 nm (47%) for the interstitial and 1522 nm (44%) for the centromeric region. A small percentage of nuclei shows band signals beyond the nuclear border. This is probably due to weak nuclear DAPI stain measured by the software eADS (see Discussion). Additional file 13: Mean radial distribution of the centromeric, interstitial and telomeric mBAND FISH signals in A PreB and B T38wt cell nuclei. The 3D FISH signal distance to the nuclear surface was measured nm using eADS software (Küpper et al., 2007) (n = number of nuclei, nm = nanometer). A In PreB cells the mean radial position of the telomeric segment was at 2016 nm and 42% relative distance from the nuclear surface, the interstitial segment at 2178 nm (46%) and the centromeric segment at 2151 nm (45%). 4. Boyle S, Gilchrist S, Bridger JM, Mahy NL, Ellis JA, Bickmore WA: The spatial organization of human chromosomes within the nuclei of normal and emerin-mutant cells. Hum Mol Genet 2001, 10(3):211–219. References Sutherland H, Bickmore WA: Transcription factories: gene expression in unions? Nat Rev Genet 2009, 10(7):457–466. 31. 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https://openalex.org/W3213737630	https://www.matec-conferences.org/articles/matecconf/pdf/2021/18/matecconf_iceaf2021_03002.pdf	English	null	On the phenomenon of impact	MATEC web of conferences	2,021	cc-by	2,625	* Corresponding author: stephan.kalapis@sla.rwth-aachen.de © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). Stephan Kalapis1, Athanasios Dafnis1, and Kai-Uwe Schröder1 Stephan Kalapis1, Athanasios Dafnis1, and Kai-Uwe Schröder1 1Institute of Structural Mechanics and Lightweight Design of RWTH Aachen, Wüllnerstraße 7 52062 Aachen, Germany 1Institute of Structural Mechanics and Lightweight Design of RWTH Aachen, Wüllnerstraße 7 Aachen, Germany Abstract. Aerostructures such as plates are commonly used in aircraft structures due to their advantageous weight to stiffness ratio. Plates are susceptible to lateral forces that occur during impact. Such impact events are caused by falling tools, bird strikes, collisions of objects, and more. Several researchers have studied impact events, but not yet the behaviour of the plate and impactor shortly after impact due to various factors. In this paper, the semi-analytical approach of Karas is used to calculate key parameters for the analysis of an impact. Karas derived this approach to calculate an impact for a simply supported plate that is impacted in the center. Here, this approach is extended to calculate the impact event at any location on the plate. The linear semi-analytical approach is written in a Python script to obtain fast results. In a code-to-code analysis, this Python code is verified with a commercial FE solver. Various parameters are varied and investigated to analyze and describe the impact phenomenon. In addition to the influence of the initial velocity of the impactor, the mass ratio of the two bodies and the impact location are also investigated. The analysis shows that the contact force and maximum displacement during an impact varies depending on the impact location, while all other parameters are kept constant. Depending on the impact location, the contact force between impactor and plate can be nine times higher than the lowest value. MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 On the phenomenon of impact Stephan Kalapis1, Athanasios Dafnis1, and Kai-Uwe Schröder1 1 Introduction Damage due to impact events are a threat to CFRP and metal plates. The hereby induced damage is often not noticed or is not visible with bare eyes. These damages may lead to failure of the overall structure. To prevent such failures, the impact event must be completely understood, particularly which variables have the greatest influence. With this better understanding of plate behaviour, the prediction of damage before it occurs is possible. There has been extensive research in the area of low velocity impact. Although such impact events are highly dynamic, Karas was able in 1939 to present an analytical solution to determine contact force and deflection of the plate for a simply supported isotropic plate while it is hit at the center [1]. This approach neglects the effect of membrane and shear forces. Karas’ solution can be considered as exact as long as the outer boundary condition meet the physical criteria which are assembled in his theory. Thus, his work is used by other researchers to validate their own work [2-3]. Further detailed research activities on impact events were conducted by Olsson. He distinguished the impact into small and large mass impact events [4]. While small mass MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 impacts tend to have peak impact forces with trailing plate deflections, the impact force and plate deflection of large mass impacts are in phase. This can be seen in Figure 1. impacts tend to have peak impact forces with trailing plate deflections, the impact force and plate deflection of large mass impacts are in phase. This can be seen in Figure 1. Fig. 1. Schematic differentiation between small and large mass impact. Fig. 1. Schematic differentiation between small and large mass impact. The small mass impact occurs if the impactors mass is lighter than 0.29 times the plates mass, and a large mass impact occurs if the impactors mass exaggerates the plates mass twice. Thus it is worth noting, that two impacts with the same kinetic energy but different masses and velocities may lead to different damages in a plate. Within this paper, the initiated damage in a plate is considered to be dependent directly by the contact force. Thus, higher contact forces between impactor and plate leads to bigger damaged areas and lower contact forces to smaller damaged areas, respectively. 2 Methodology In order to calculate the impact on arbitrary position analytically, the work of Karas is chosen to be further investigated. With the analytical solution it is possible to calculate the dynamic deflection of impactor and plate during an impact event as well as the corresponding contact force between them. As seen in figure 2, the plate is considered to have the width a, length b and depth h. The impactor has the mass mg and radius r, while hitting the plate with the velocity v0. Plate and impactor are made out of the same isotropic material with Youngs Modulus E, Poisson’s Ratio ν and density ρ. As the impact location is considered to be arbitrary, this location is described by ξ and η. 2 2 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 Fig. 2. Plate and impactor dimension. Fig. 2. Plate and impactor dimension. Fig. 2. Plate and impactor dimension. Karas derived the following equation, which is based on the Kirchhoff hypothesis, to calculate the plate deflection w [1]: (, ) = ∑ ∑ ∫() sin + ( −)! " $ % × sin & sin ' (1) (1) (1) with with = - (./) (2) (2) (2) Where m, n are even numbers and t the considered time. The expression Qmn is the generalized force. In his work, Karas set the generalized force to be the contact force P. This expression is here developed to be () = 0() sin 1 sin 2 (3) (3) In contrast to Karas, who focused on an analytical formulation, this variant is concerned with the arbitrariness of the impact location. As mentioned above, this paper is not about the formulation of a new analytical approach, but about the effects of different impact locations. As P and w in Eq. 3 are unknown, another equation is needed to substitute the deflection w. As the impactor intendates the plate, a Hertzian Contact is obtained. This indentation and the plate deflection w must yield the overall travelled distance u of the impactor. 2 Methodology While the indentation is calculable by the Hertzian Contact law, the position of the impactor is determined by following equation: 3% − ∫0()( −)" $ 4 (4) 3% − ∫0()( −)" $ 4 (4) (4) Together with Eq. 3, Eq. 4 and the contact law, one is able to calculate the impact force P by numeric integration. To determine the plates deflection, Eq. 1 must be solved. Here, this routine is automated via a Python-Script. Within this script, it is possible to change the evaluation location x, y independently from the impact location ξ and η. This approach considers only linear bending of the plate as Kirchhoff hypothesis for bending plates is included, while non-linear effects such as membrane or shear forces are neglected. 3 3 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 4 Analytical Investigation and Discussion As discussed in section 1, it is important to distinguish between a small mass and a large mass impact. They may lead to different contact forces and thus to different damage-affected areas. Moreover, it is worth knowing if and how these contact forces differ on arbitrary plate location. For this purpose, a small mass and a large mass impact are studied on arbitrary impact locations. For both types of impact the kinetic energy equals 10 J. The initial velocity is set to fulfil the given mass and level of kinetic energy. The investigated plate is the same as in section 3, thus it has a dimension of 200 x 200 x 8 mm³ and is made out of steel with Young’s Modulus of 251,820 N/mm² and Poissons Ratio of 0,3. An isotropic material is choosed as Eq. 1 can only handle such materials. For investigations on anisotropic materials such as CFRPs, a quasi-isotropic laminate must be chosen. In figure 4 the maximum deflection of the plate, the maximum contact force and the number of subsequent impacts in such an event is shown for both types of impact. The calculated time was chosen to guarantee that no contact between plate and impactor is missed. Fig. 4. Evaluation of plate deflection, contact force and number of impacts for small and large mass impact on arbitrary position. f ll d l Fig. 4. Evaluation of plate deflection, contact force and number of impacts for small and large mass impact on arbitrary position. Fig. 4. Evaluation of plate deflection, contact force and number of impacts for small and large mass impact on arbitrary position. On the first look it is evident, that the deflection and force distribution over the plate are not equal for large and small mass impact. Although, as expected, the deflection decreases approaching the boundary while the contact force increases. This effect leads back to the higher plate stiffness near the boundary and no much movement of the plate is observed. This explanation is backed by the fact, that in the boundary region only one contact during the impact event occurs. For the small mass impact, more dynamic events are observed in the middle of the plate. Here, two to four single contacts are occurring leading to diverging maximum deflection of the plate. 3 Numerical verification To verify the derived equation, the obtained results are compared with a numeric finite element analysis (FEA). The program used is Abaqus, Dassault Systems. A plate with the dimensions of 200 x 200 x 8 mm³ is impacted by a sphere with radius 10 mm and an initial velocity of 1000 mm/s. Both plate and impactor are made out of steel with Youngs Modulus of 251,820 N/mm² and Poissons Ratio of 0,3. As seen in figure 3, three different locations are evaluated: mid, edge and corner. Following figure 2, the coordinates for the impacts are (100 mm \| 100 mm) for mid, (50 mm \| 100 mm) for edge and (50 mm \| 50 mm) for corner. The simply supported plate consists of 1,156,000 C3D8R elements, the impactor consists of 107,200 C3D8R elements. While the plate deflection can be measured in the post-processing, the impact force must be calculated separately. Therefore, the average acceleration of the impactor is extracted and multiplied by its mass. Using Newtons Second Law the impact force is now obtained. Fig. 3. Verification of the proposed model with FEA. Impact occurs a) in the mid, b) in the side and c) in the corner of the plate. Fig. 3. Verification of the proposed model with FEA. Impact occurs a) in the mid, b) in the side and c) in the corner of the plate. Following figure 3, the calculated plate deflection and the contact force between plate and impactor of the Python-Script matches the FEA well, regardless the impact location. Hence, there are some minor differences between proposed model and FEA: a slight shift for the impact force and diverging deflections of the plate to the end of the impact event. This circumstance can be justified by the non-linear calculation of the FEA, while the proposed model uses linear approaches. Similar differences were reported by other authors as well [5- 6]. Furthermore, it was verified that the results are perfectly symmetric along x- or y- axis or the diagonal thereof. Thus, the numerical Python-Script is found to work correctly. 4 4 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 4 Analytical Investigation and Discussion Interestingly, the maximum contact force is not much affected in the middle of the plate: from the center to the boundary the contact forces decreases a little bit and then it raises to the boundary. The large mass impact behaves the opposite way than the small mass impact. Here are mainly one, at some regions in circles around the center two, contacts in the impact event. While the maximum deflection reduces from the center to the boundary as expected, the 5 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 MATEC Web of Conference 349, 03002 (2021) ICEAF-VI 2021 https://doi.org/10.1051/matecconf /202134903002 contact force has some local abnormalities. Around the center exists a circle, where the contact force is dropping. Around the area at (0.68 \| 0.68) a local maximum is observed. contact force has some local abnormalities. Around the center exists a circle, where the contact force is dropping. Around the area at (0.68 \| 0.68) a local maximum is observed. Figure 4 shows the high dependency of the location on impact events regarding the plates deflection and contact force. If the impact event happens just 10 mm further away, a different damage area could be observed. Although the derived equations are only valid for thin plates and the here investigated plate does not fit that criteria well, the here obtained results can be used for a first rough estimation. The influence of shear or membrane forces is considered to be negligible in this case. 5 Conclusion In this paper the impact event of a round sphere on a flat plate is investigated. The analytical approach of Karas was expanded to calculate impact events on arbitrary plate positions by using a more detailed approach for the generalized force. The here analytical derived approach was compared and verified with a commercial FEA. Here, it was shown that the impact location has a tremendous influence on the impact event, regarding its plate deflection and contact force history. Thus, it appears that some areas on a plate are more susceptible to damages by impacts. These areas must not necessarily lay in the middle of the plate but a little bit outside of it. Furthermore, the mass ratio influences the position of this area. A general pattern, where the maximum contact force can be expected is not given here, though it must be further researched. Focus in further research must be given on the detailed dynamics of the impact and the here involved parts. The driving variables must be identified, which will lead to the differentiation of small and large mass impact. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References [2] S. Khalili, K. Malekzadeh, A. V. Gorgabad, Compos. Struct. 96 (2013) 64–74. doi:10.1016/j.compstruct.2012.07.035. [2] S. Khalili, K. Malekzadeh, A. V. Gorgabad, Compos. Struct. 96 (2013) 64–74. doi:10.1016/j.compstruct.2012.07.035. [3] D. Chakraborty, Mater. Des. 28 (4) (2007) 1142–1153. doi:10.1016/j.matdes.2006.01.029. [3] D. Chakraborty, Mater. Des. 28 (4) (2007) 1142–1153. doi:10.1016/j.matdes.2006.01.029. [4] R. Olsson, Compos. Part A Appl. Sci. Manuf. 31 (8) 490 (2000) 879–887. doi:10.1016/S1359-835X(00)00020-8. [4] R. Olsson, Compos. Part A Appl. Sci. Manuf. 31 (8) 490 (2000) 879–887. doi:10.1016/S1359-835X(00)00020-8. ( ) [5] M. R. Bayat, O. Rahmani, M. Mosavi Mashhadi, Polym. Compos. 39 (3) (2018) 730– 745. doi:10.1002/pc.23990. [5] M. R. Bayat, O. Rahmani, M. Mosavi Mashhadi, Polym. Compos. 39 (3) (2018) 730– 745. doi:10.1002/pc.23990. [5] M. R. Bayat, O. Rahmani, M. Mosavi Mashhadi, Polym. Compos. 39 (3) (2018) 730– 745. doi:10.1002/pc.23990. [6] M. Ghalami-Choobar, G. Liaghat, M. Sadighi, H. Ahmadi, Compos. Struct. 201 (2018) 315–325. doi:10.1016/j.compstruct.2018.06.027. [6] M. Ghalami-Choobar, G. Liaghat, M. Sadighi, H. Ahmadi, Compos. Struct. 201 (2018) 315–325. doi:10.1016/j.compstruct.2018.06.027. [6] M. Ghalami-Choobar, G. Liaghat, M. Sadighi, H. Ahmadi, Compos. Struct. 201 (2018) 315–325. doi:10.1016/j.compstruct.2018.06.027. 6
https://openalex.org/W2995963179	https://revistas.uneb.br/index.php/tabuleirodeletras/article/download/7727/5047	Portuguese	null	Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA)	Tabuleiro de Letras	2,019	cc-by	5,843	2 Universidade Federal do Pará / CNPq. E-mail: reginafcruz@gmail.com 1 Universidade Federal do Pará. E-mail: msebastcosta@gmail.com 1 Universidade Federal do Pará. E-mail: msebastcosta@gmail.com 2 Universidade Federal do Pará / CNPq. E-mail: reginafcruz@gmail.com Este é um artigo publicado em acesso aberto (Open Access) sob a licença Creative Commons Attribution, que permite uso, distribuição e reprodução em qualquer meio, sem restrições desde que o trabalho original seja corretamente citado. Entoacional analysis in the Amazon: an acoustic study in Portuguese spoken in Mocajuba and Cametá (PA) Maria Sebastiana da Silva Costa1 https://orcid.org/0000-0001-7918-1972 Maria Sebastiana da Silva Costa1 https://orcid.org/0000-0001-7918-1972 Regina Célia Fernandes Cruz2 https://orcid.org/0000-0003-3985-1024 Resumo: Este trabalho compreende um estudo comparativo das descrições prosódicas prévias sobre as variedades de Mocajuba e Cametá (PA) no seio do projeto AMPER-POR (COSTA, em andamento). Visa-se à descrição do comportamento entoacional de sentenças nas modalidades declarativa neutra e interrogativa total. Os parâmetros acústicos de frequência fundamental (doravante F0), Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) duração e intensidade foram foco de análise. Os procedimentos metodológicos adotados foram previamente estabelecidos pelo projeto AMPER-POR (CRUZ et al., 2012). Os movimentos melódicos das curvas de F0, nas modalidades frasais interrogativas e declarativas, para a variedade de Cametá, registraram menos variação intersilábica de F0, em relação a Mocajuba, indicando, portanto, essa variedade como tendo maior musicalidade, nos três tipos de acento lexical. Desta forma, apenas os resultados concernentes à variação melódica de F0 da variedade falada em Mocajuba assemelha-se com padrões entoacionais encontrados para outras regiões brasileiras, mais precisamente para o Norte e Nordeste brasileiro conforme descrito por Moraes (2003 apud SILVA e CUNHA, 2012, p. 10) e Silva e Cunha (2012). O parâmetro acústico de duração apresentou uma distinção dialetal significativa, posto que os dados de Cametá registraram sílabas mais longas, em relação a Mocajuba. A intensidade não apresentou variação significativa entre os dialetos estudados. duração e intensidade foram foco de análise. Os procedimentos metodológicos adotados foram previamente estabelecidos pelo projeto AMPER-POR (CRUZ et al., 2012). Os movimentos melódicos das curvas de F0, nas modalidades frasais interrogativas e declarativas, para a variedade de Cametá, registraram menos variação intersilábica de F0, em relação a Mocajuba, indicando, portanto, essa variedade como tendo maior musicalidade, nos três tipos de acento lexical. Desta forma, apenas os resultados concernentes à variação melódica de F0 da variedade falada em Mocajuba assemelha-se com padrões entoacionais encontrados para outras regiões brasileiras, mais precisamente para o Norte e Nordeste brasileiro conforme descrito por Moraes (2003 apud SILVA e CUNHA, 2012, p. 10) e Silva e Cunha (2012). O parâmetro acústico de duração apresentou uma distinção dialetal significativa, posto que os dados de Cametá registraram sílabas mais longas, em relação a Mocajuba. A intensidade não apresentou variação significativa entre os dialetos estudados. Palavras-chave: Descrição Prosódica; Projeto AMPER-POR; Análise Acústica. 3 http://www.varialing.eu/ Entoacional analysis in the Amazon: an acoustic study in Portuguese spoken in Mocajuba and Cametá (PA) Abstract: This work comprises a comparative study of previous prosodic descriptions from the varieties of Mocajuba and Cametá (PA) within the AMPER-POR project (COSTA, in progress). It aims to describe the intonation behavior of sentences in the neutral declarative and total interrogative modalities. The acoustic parameters of fundamental frequency (hereinafter F0), duration and intensity were the focus of analysis. The methodological procedures adopted were previously established by the AMPER-POR project (CRUZ et al., 2012). The melodic movements of the F0 curves, in the interrogative and declarative phrasal modalities, for the Cametá variety, registered less inter-syllabic variation of F0 in relation to Mocajuba, indicating, therefore, this variety as having greater musicality in the three lexical accent types. Thus, only the results concerning the melodic F0 variation from the variety spoken in Mocajuba resembles intonation patterns found for other Brazilian regions, more precisely for the North and Northeast of Brazil as described by Moraes (2003 apud SILVA and CUNHA, 2012, p. 10) and Silva and Cunha (2012). The acoustic duration parameter presented a significant dialectal distinction, since Cametá data recorded longer syllables in relation to Mocajuba. The intensity did not present significant variation among the dialects studied. Keywords: Prosodic Description; AMPER-POR Project; Acoustic analysis. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. Keywords: Prosodic Description; AMPER-POR project; Acoustic Analysis. Palavras-chave: Descrição Prosódica; Projeto AMPER-POR; Análise Acústica. Introdução A prosódia do Português Brasileiro (PB) tem sido alvo de investigação recente pelo projeto AMPER-POR3, neste sentido, tem-se realizado estudos de natureza prosódica com análises pautadas na entoação de frases. Por conseguinte, este artigo objetiva a descrição prosódica acústica de dados do projeto AMPER-POR para o estado do Pará, especialmente, em sentenças declarativas neutras e interrogativas totais do português falado em Cametá (PA) e Mocajuba (PA). Para tanto, foi realizada uma análise interdialetal, a fim de caracterizar o padrão entoacional do falar cametaense e mocajubense por meio de três parâmetros acústicos: Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 53 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Frequência Fundamental (medida em Hz), Duração (medida em ms) e Intensidade (medida em dB), a fim de responder questões, como: o padrão entoacional encontrado para as frases na modalidade declarativa neutra falada por locutores da cidade de Cametá é similar ao encontrado para Mocajuba? O padrão entoacional encontrado para as interrogativas da cidade de Cametá é semelhante ao encontrado para Mocajuba? Os padrões entoacionais encontrados para as declarativas neutras e interrogativas totais são os mesmos encontrados para outras regiões do país? Os parâmetros de duração (ms) e intensidade (dB) são fatores determinantes de distinção nas variedades em escopo? Este artigo pauta-se na teoria da Sociofonética, uma ciência nova que possui por objeto de estudo a aglutinação de dois campos de investigação mais antigos, a Fonética e a Sociolinguística, como toda investigação conduzida no seio do projeto AMPER-POR. Isto posto, este trabalho trata das variações fonéticas acústicas e as características sociais dos falantes, como escolaridade e dialeto. Entre alguns teóricos que se encarregam deste estudo, cita-se: Foulkes e Scobbie e Watt (2010), Baranowski (2013) e Hay e Drager (2007 apud SORIANO, 2016). A metodologia adotada é a mesma preestabelecida pelo projeto AMPER-POR4 (CRUZ et al., 2012) e para este estudo comparativo foram analisadas sentenças correspondentes as três pautas acentuais do Português: oxítona, paroxítona e proparoxítona, todas na fala de informante do sexo feminino, sendo o alvo da análise o último vocábulo do Sintagma Nominal Final de sentenças selecionadas previamente. 4 Atlas Prosódico Multimédia do Espaço Românico. Introdução Em vistas gerais, este trabalho divide-se nas seguintes seções: Fundamentação Teórica (seção 1), em seguida, a descrição dos procedimentos metodológicos adotados para a concretização deste trabalho (seção 2), seguido da apresentação dos resultados (seção 3) e suas considerações finais. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 1 Fundamentação teórica Segundo Foulkes e Scobbie e Watt (2010), a palavra Sociofonética surgiu em 1974, na Dissertação de Deshaies-Lafontaine, a qual tratava sobre a variação do Francês canadense. O intuito da utilização do termo foi para demonstrar a conformidade entre as ciências Sociolinguística e Fonética em detrimento da Sintaxe ou do Léxico. De acordo com Foulkes e Scobbie e Watt (2010), a Sociofonética é um campo de 54 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) investigação linguística que envolve a integração de princípios, técnicas e quadros teóricos da Fonética e da Sociolinguística, com a finalidade de explicar a variação social da fala. Para Baranowski (2013), a Sociofonética é um instrumento que contribui para a compreensão da natureza da variação linguística e mudança, deste modo, é uma ciência que estuda como a língua falada varia de acordo com o contexto social em que ocorre, e como falantes e ouvintes são sensíveis a esta variação, portanto, está atrelada à produção e à percepção da fala em variação. Como bem tem demonstrado estudos neste campo, a realização de qualquer palavra pode variar de acordo com o falante, o idioma, o contexto social, o tópico, o destinatário, a postura intencional tomada pelo orador, e inúmeros outros fatores. Neste contexto, a identidade linguística é construída simultaneamente a informações sociais, conforme explicita Soriano (2016). Isto posto, a Fonética Acústica, que se encarrega de estudar aspectos físicos da produção da fala, tem demonstrado que o fenômeno da variação é um campo de estudo desta ciência e que estes fenômenos são provenientes, na maioria dos casos, não somente de fatores linguísticos, mas também de fatores sociais. Este trabalho valeu-se da teoria da Sociofonética, exatamente por ser constituída de métodos baseados na Fonética Acústica e na Sociolinguística, haja vista a necessidade em considerar como fatores de análise, respectivamente, o parâmetro acústico fonético de F0 e fatores sociais, como procedência e escolaridade dos locutores entrevistados para se encontrar um padrão entoacional das localidades em escopo. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 2 Estudos geoprosódicos do PB Silva e Cunha (2012) caracterizaram o padrão entoacional de vinte e cinco capitais brasileiras. Para o trabalho intitulado “Caracterização prosódica dos falares brasileiros: a oração interrogativa total” que está vinculado ao projeto Atlas Linguístico do Brasil (ALiB), as autoras analisaram a variedade de cinco capitais: Teresina, João Pessoa, Maceió, Cuiabá e Florianópolis. As análises incidiram em sentenças na modalidade frasal, interrogativa total e procuraram responder, entre outras questões: 1) Quais são as implementações fonéticas dos acentos nucleares do sintagma entoacional da questão total nos falares das capitais brasileiras? 2) Qual é a distribuição geográfica dos padrões regionais da questão total no PB? Para tanto, foram inquiridos quatro informantes não escolarizados por capital, distribuídos em duas faixas etárias 18 a 30 anos e 50 a 65 anos e nos dois sexos, feminino e 55 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) masculino. Todos nascidos e criados na localidade pesquisada. O questionário aplicado segue o modelo do questionário fonético-fonológico (QFF) do ALiB. As autoras afirmam que a diferença regional está principalmente no acento nuclear. Elas confirmam que o comportamento entoacional ascendente-descente no acento nuclear descrito por Moraes (2008) através da notação fonológica L+H*L% está presente em praticamente todas as capitais brasileiras, com exceção de João Pessoa. Um outro trabalho relevante que serviu para respaldar nossas análises foi o de Reis e Cunha (2012). As autoras estudaram o comportamento prosódico dialetal do português falado na capital de São Luis (MA) e no município de Brejo (MA). Suas análises foram substanciadas no campo da fonologia entoacional, entre os principais teóricos citados estão Sosa (1999), Cunha (2011) e Moraes (2003). Alguns resultados encontrados para a caracterização entoacional do português brasileiro são citados pelas autoras, entre eles, o padrão entoacional para as declarativas, na região nuclear da sentença, relativo às variedades de Recife e Porto Alegre. Para Recife o movimento é identificado com um tom alto na sílaba pretônica, sofrendo uma descendência nas sílabas tônicas e postônicas. Por outro lado, para Porto Alegre, o movimento melódico inicia com tom alto na sílaba pretônica, espraiando-se até a sílaba tônica, para posteriormente sofrer descendência na postônica. Reis e Cunha (2012, p. 11) afirmam que Moraes (2003) apresenta um notação para a modalidade interrogativa. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 2 Estudos geoprosódicos do PB A descrição do movimento de F0 registra uma subida melódica na sílaba pretônica até a sílaba tônica para posteriormente sofrer descendência na sílaba postônica final. Os resultados relacionados a modalidade interrogativa no estudo realizado por Reis e Cunha (2012) registraram dois contornos melódicos, na região nuclear da sentença. Para a cidade de Brejo, o contorno melódico do último vocábulo da sentença inicia com um desenho de ascendência da sílaba pretônica até a sílaba tônica, mantendo-se com tom alto na postônica. Por outro lado, para São Luis, o movimento apresenta o desenho ascendente-descendente, com pico entoacional na sílaba tônica do vocábulo. Logo os resultados encontrados por Reis e Cunha (2012) para duas cidades do Maranhão registrou variação dialetal entoacional. As pesquisas realizadas por Reis e Cunha (2012) e Silva e Cunha (2012) serviram para consubstanciar nosso trabalho, haja vista que se enquadram em estudos que também buscam descrever a variação entoacional, em várias regiões do Brasil. Para além, os resultados encontrados pelas autoras serviram para a interpretação de nossas análises no correspondente a variação melódica de F0 na variedade mocajubense. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 56 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 3 Metodologia: formação e organização do corpus de Cametá (PA) e Mocajuba (PA) Adotaram-se, no presente estudo, todos os procedimentos metodológicos determinados pela coordenação geral do projeto AMPER-POR (CRUZ et al., 2012), e como um dos objetivos do projeto compreende uma análise contrastiva dos dialetos estudados, o corpus gravado é formado por seis repetições de 102 frases do corpus de base do projeto para a Língua Portuguesa. Cada um dos elementos constituintes das frases possui uma imagem correspondente, uma vez que não é permitido nenhum contato dos informantes com as frases escritas. No momento da coleta de dados, a cada informante, foram pedidas seis repetições da série de frases do corpus (em ordem aleatória), ao todo, são geradas 612 repetições, sendo selecionadas para análise acústica as três melhores repetições, a fim de serem estabelecidas médias dos parâmetros acústicos: Frequência fundamental (em Hz), Duração (em ms) e Intensidade (em dB). Sintaticamente, as frases são montadas de forma a apresentar Sujeito- Verbo-Complemento (SVC) e suas expansões com a inclusão de Sintagmas Preposicionais. Elas são concebidas de modo a contemplar as modalidades declarativa neutra e interrogativa total. Uma descrição mais detalhada da coleta dos dados, levantamento do corpus do trabalho e seleção dos informantes pode ser encontrada em Costa (2015, p. 54-58). O material gravado passou, então, por seis etapas de tratamento: a) codificação; b) isolamento das repetições em arquivos de áudios individuais; c) aplicação do script lance_batch_easyalign_v3.praat para obter o textgrid dos arquivos .wav; d) segmentação fonética no programa PRAAT 6.0.39 por meio de um script de correção_segmentação.praat para corrigir a segmentação; e) aplicação do script AMPER_Textgrid2Txt_V3_boucle_DepoisEasyAlign_v2.praat para gerar os arquivos com os parâmetros acústicos; f) seleção das três melhores repetições; g) rodadas no programa Matlab para se obter os arquivos fonos, estes que fornecem as médias de F0, duração e intensidade necessárias para o preenchimento das planilhas Excel e, consequentemente, para a feitura dos gráficos. De posse dos arquivos fonos gerados em arquivos .TXT, procedeu-se à feitura dos gráficos no Excel. Para tanto, preencheu-se planilhas com as medidas dos parâmetros físicos analisados do último vocábulo da sentença, acompanhado da partícula “do”, os quais estão 57 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) distribuídos nos três tipos de acento lexical, respectivamente, proparoxítono: “do pássaro”, paroxítono: “do Renato” e oxítono: “do bisavô”, confrontando sempre a variável dialetal, em vocábulos declarativos e interrogativos. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 3 Metodologia: formação e organização do corpus de Cametá (PA) e Mocajuba (PA) Os gráficos contendo as medidas de duração e intensidade foram montados, considerando, apenas, a média desses parâmetros na sílaba tônica dos vocábulos alvo, ou seja foram tomadas as medidas de duração e intensidade da sílaba ‘pá’ (v3) dos vocábulos proparoxítonos, da sílaba ‘na’ (v2) dos vocábulos paroxítonos e da sílaba ‘vô’ (v1) dos vocábulos oxítonos. Foram selecionadas duas frases de cada pauta acentual, conforme apresentado no quadro 1. Quadro 1 – Especificação do corpus selecionado para análise ACENTUAÇÃO (CÓDIGO/AMPER) MOCAJUBA CAMETÁ Oxítona (kwk) O bisavô gosta do bisavô O bisavô gosta do bisavô Oxítona (pwk) O pássaro gosta do bisavô O pássaro gosta do bisavô Paroxítona (kwt) O bisavô gosta do Renato O bisavô gosta do Renato Paroxítona (pwt) O pássaro gosta de Renato O pássaro gosta de Renato Proparoxítona (pwp) O pássaro gosta do pássaro O pássaro gosta do pássaro Proparoxítona (kwp) O bisavô gosta do pássaro O bisavô gosta do pássaro Elaboração: Da autora, 2019 Desta maneira, foram analisadas as três melhores repetições de seis sentenças na modalidade declarativa neutra e seis na modalidade interrogativa total, na fala de seis locutores do sexo feminino, pertencentes as duas variedades em escopo, três mulheres de Mocajuba e três mulheres de Cametá, dos níveis fundamental, médio e superior de escolaridade, totalizando um corpus de 432 repetições. A análise, como já mencionado, incidiu na variação dos parâmetros acústicos de F0, duração e intensidade – controlando o sintagma nominal final do enunciado. Foram calculadas Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 58 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) as médias de cada parâmetro físico controlado. As frases escolhidas apresentam a mesma estrutura frasal, do tipo SVC, todas com dez vogais, e compostas por vocábulos proparoxítonos, paroxítonos e oxítonos que ocupam a posição de sujeito e complemento verbal (objeto). Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados As análises respaldam-se no núcleo acentual do sintagma nominal final das frases, pelo qual se busca comparar as variáveis de procedência e incidiram na região nuclear da sentença, pois, como afirma Moraes (2008 apud SILVA; CUNHA, 2012), a identificação de uma frase interrogativa se dá pela movimento ascendente que ocorre, preferencialmente, no acento nuclear da frase fonológica, ou seja, no último vocábulo da sentença. As autoras ainda ressaltam a importância dessa subida melódica na distinção entre uma frase declarativa neutra e interrogativa total. Para análise foi considerado o acento lexical tônico do vocábulo-alvo, que ora pode incidir na sílaba v3, se o vocábulo analisado for proparoxítono, pássaro; ora na sílaba v2, se o vocábulo analisado for paroxítono, Renato; e ora na sílaba v1, se o vocábulo analisado for oxítono, bisavô. Todos os vocábulos foram antecedidos pela partícula preposicional, “do” que ocupará sempre a posição, nos gráficos, de v4. As interpretações consideraram as sílabas adjacentes, pretônicas; e as subjacentes, postônicas às sílabas tônicas. Nas figuras abaixo, lê-se o contorno prosódico encontrado para todas as frases de mesma estrutura acentual (tipo de acento) nas modalidades frasais alvo, a fileira de cima, representa sempre as frases declarativas neutras; e a fileira de baixo, as interrogativas totais. 59 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Figura 1 – Gráficos das variações de F0 nas quatro sílabas finais de enunciado por variedade: Cametá (azul) e Mocajuba (vermelho), em três tipos de acento lexical (respectivamente, proparoxítono, paroxítono e oxítono) e escolaridade fundamental Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Conforme observado na figura acima, referentes aos dados dos locutores do nível mais baixo de escolaridade do corpus (Ensino Fundamental), os contornos melódicos de F0, nas duas variedades em estudo, apresentaram distinção melódica intersilábica, nas declarativas neutras proparoxítona e paroxítona (fileira superior), enquanto que nas interrogativas total (fileira inferior) o movimento diferenciou-se apenas nas oxítonas. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados O movimento de F0 significativo que estabelece a distinção das duas modalidades alvo, caracterizado como ascendente em enunciados interrogativos e descendente para os enunciados declarativos, ocorre justamente na 60 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) sílaba tônica do vocábulo, independente do tipo de acento lexical, respectivamente, nas sílabas proparoxítonas (v3), paroxítonas (v2) e oxítonas (v1), apresentando uma maior inclinação das curvas melódicas de F0 na fala dos mocajubenses, nas duas modalidades em escopo. Apesar de o pico maior de ascendência de F0, característico dos enunciados interrogativos, ocorrer na sílaba tônica de cada vocábulo, o movimento é “desenhado” na sílaba pretônica. Nas proparoxítonas acontece da sílaba pretônica v4 (do), registrando queda continua de F0 até a sílaba postônica final v1; nas paroxítonas na sílaba pretônica v3 (Re) e atingiu seu pico na sílaba tônica v2 (na), registrando queda de F0 na sílaba postônica v1 (to); e nas oxítonas na sílaba pretônica v2 (sa) com ascendência constante até a sílaba tônica v1 (vô). Comportamento similar de F0 é registrado nas declarativas, uma vez que, também, é na sílaba pretônica que ocorre o pico de F0 necessário para antecipar o movimento de descendência de F0 característicos da entoação declarativa. Nos vocábulos proparoxítonos, o pico de F0 ocorre na sílaba pretônica v4 (do) para ocorrer a queda na sílaba tônica (pá); nas paroxítonas na sílaba pretônica v3 (Re) e imediatamente vai caindo até a sílaba postônica v1 (to), nas oxítonas na sílaba pretônica v2 (sa) caindo até a sílaba tônica v1 (vô). A variedade de Mocajuba acompanha a descrição prevista para as declarativas do PB, uma vez que o movimento melódico das curvas é o mesmo descrito por Moraes (2003 apud SILVA e CUNHA, 2012, p. 10) quando se refere as frases declarativas para o Português do Brasil, ao afirmar que as declarativas, de um modo geral, apresentam tom baixo na tônica que se espraia à postônica. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados 61 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Figura 2 – Gráficos das variações de F0 nas quatro sílabas finais de enunciado por variedade: Cametá (azul) e Mocajuba (vermelho), em três tipos de acento lexical, em três tipos de acento lexical (respectivamente, proparoxítono, paroxítono e oxítono) e escolaridade média Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Nos gráficos correspondentes ao Ensino Médio, nota-se, que os contornos melódicos, das variedades em estudo, apresentam uma distinção do tipo de movimento de F0 nos vocábulos referentes às pautas acentuais proparoxítona, paroxítona e oxítona, nas declarativas. Para as interrogativas, observou-se essa variação nas proparoxítonas e paroxítonas. Apesar de em ambas as variedades o movimento distintivo de F0 ocorrer na sílaba tônica das duas modalidades alvo, caracterizando descendência nas declarativas e ascendência nas interrogativas, registram-se valores mais altos nas sílabas contendo o pico de F0 na variedade de Mocajuba. Nas declarativas de Mocajuba, o movimento melódico inicia na sílaba pretônica de cada vocábulo, com movimento de descendência nas sílabas tônicas e postônicas, com a exceção das proparoxítonas, para as quais não foi possível encontrar um padrão entoacional. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 62 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Nas paroxítonas a F0 atingiu seu pico máximo na sílaba pretônica v3 (Re) registrando uma queda melódica na sílaba tônica v2 (na); e nas oxítonas na sílaba pretônica v2 (sa) com ascendência constante até a sílaba tônica v1 (vô). Nas paroxítonas a F0 atingiu seu pico máximo na sílaba pretônica v3 (Re) registrando uma queda melódica na sílaba tônica v2 (na); e nas oxítonas na sílaba pretônica v2 (sa) com ascendência constante até a sílaba tônica v1 (vô). A descrição das curvas melódicas das frases interrogativas nas paroxítonas, assemelha- se à feita por Silva e Cunha (2012) para a região Norte e Nordeste do Brasil, na qual ocorre uma subida melódica da F0 na sílaba tônica do acento nuclear, que se estende à postônica final, caracterizando um movimento ascendente nas interrogativas, o movimento foi melhor observado nas paroxítonas da variedade de Mocajuba. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados Figura 3 – Gráficos das variações de F0 nas quatro sílabas finais de enunciado por variedade: Cametá (azul) e Mocajuba (vermelho), em três tipos de acento lexical (respectivamente, proparoxítono, paroxítono e oxítono) e escolaridade superior Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Legenda: Cv.cv.cv (proparoxítono), cv.Cv.cv (paroxítono) e cv.cv.Cv (oxítono) Elaboração: Da autora, 2019 Os contornos melódicos nos gráficos acima são correspondentes aos dados de fala das locutoras do Ensino Superior das variedades analisadas. Registrou-se um pico entoacional nas Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 63 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) sílabas pretônicas, na modalidade frasal declarativa para os três tipos de acento lexical, respectivamente na sílaba v4 (do) para as proparoxítonas, nas paroxítonas na sílaba pretônica v3 (Re) e nas oxítonas na sílaba pretônica v2 (sa) para posteriormente ocorrer uma queda de F0 nas sílabas tônicas de cada vocábulo analisado. Para as interrogativas o pico entoacional aconteceu na sílaba tônica dos três tipos de acento analisado. Para além, notou-se que houve uma distinção melódica em relação as variedades em escopo, posto que apenas Mocajuba apresentou movimentos característicos das curvas melódicas de F0, principalmente para as interrogativas, correspondentes aos resultados encontrados por Silva e Cunha (2012) como já mencionado. Notou-se movimentos mais discretos de F0 para Cametá, não demonstrando variação significativa intersilábica nas três pautas acentuais. Abaixo serão apresentadas as análises referentes à Duração, sendo confrontados os fatores escolaridade e variação dialetal. Figura 4 – Gráficos das variações de Duração (ms) na sílaba tônica de vocábulos finais de enunciado por variedade: Cametá (azul) e Mocajuba (vermelho), em três tipos de acento lexical (respectivamente, proparoxítono, paroxítono e oxítono) nas três escolaridades analisadas Legenda: Enunciados declarativos (fileira de cima) e interrogativos (fileira de baixo) e escolaridades Fundamental (EF), Médio (EM) e Superior (ES) Elaboração: Da autora, 2019 Legenda: Enunciados declarativos (fileira de cima) e interrogativos (fileira de baixo) e escolaridades Fundamental (EF), Médio (EM) e Superior (ES) Elaboração: Da autora, 2019 Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados 64 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Nos gráficos referentes ao parâmetro acústico Duração (medido em ms), notou-se que os níveis de tempo foram sempre mais elevados para a variedade falada em Cametá, enquanto Mocajuba manteve-se com níveis baixos de duração, o fenômeno ocorreu para as três pautas acentuais em estudo. Parecendo, portanto, a duração um parâmetro físico importante na caracterização dialetal das variedades em análise. Figura 5 – Gráficos das variações de Intensidade (dB) na sílaba tônica de vocábulos finais de enunciado por variedade: Cametá (azul) e Mocajuba (vermelho), em três tipos de acento lexical (respectivamente, proparoxítono, paroxítono e oxítono) nas três escolaridades analisadas Legenda: Enunciados declarativos (fileira de cima) e interrogativos (fileira de baixo), escolaridades Fundamental (EF), Médio (EM) e Superior (ES) Elaboração: Da autora, 2019 Legenda: Enunciados declarativos (fileira de cima) e interrogativos (fileira de baixo), escolaridades Fundamental (EF), Médio (EM) e Superior (ES) Elaboração: Da autora, 2019 Uma análise dos gráficos concernentes ao parâmetro acústico de Intensidade (dB) demonstrou que não houve variação relevante de energia entre a fala dos locutores das variedades em estudo, haja vista que não houve uma diferença significativa entre os valores correspondentes à escolaridade e pauta acentual. Como mencionado na introdução deste artigo, 4 eram as hipóteses a serem verificadas: Como mencionado na introdução deste artigo, 4 eram as hipóteses a serem verificadas: Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 65 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) a) se o padrão entoacional encontrado para as frases na modalidade declarativa neutra falada por locutores da cidade de Cametá é similar ao encontrado para Mocajuba; b) se o padrão entoacional encontrado para as interrogativas da cidade de Cametá é semelhante ao encontrado para Mocajuba; c) se os padrões entoacionais encontrados para as declarativas neutras e interrogativas totais são os mesmos encontrados para outras regiões do país e; d) se os parâmetros de duração (ms) e intensidade (dB) são fatores determinantes de distinção nas variedades em escopo. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. BARANOWSKI, M. Sociophonetics. In: BAYLEY, Robert; CAMERON, Richard; LUCAS, Ceil (eds.). The Oxford handbook of sociolinguistics. Oxford: Oxford University Press, 2013. p.403-424. 4 Análise e discussão dos resultados O movimento é observado nas três pautas acentuais e nos três níveis de 66 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) escolaridade analisados; escolaridade analisados; escolaridade analisados; e) O parâmetro de duração (ms) é um fator determinante de distinção nas variedades em escopo, posto que os dados de Cametá registraram uma variação de tempo que parece ser significativa em relação a Mocajuba, registrando, sempre, sílabas mais longas que as de Mocajuba; f) O parâmetro de intensidade (dB) não é um fator determinante de distinção nas variedades em escopo, posto que não houve variação de energia entre os dialetos estudados. f) O parâmetro de intensidade (dB) não é um fator determinante de distinção nas variedades em escopo, posto que não houve variação de energia entre os dialetos estudados. Considerações finais Apresentaram-se aqui os resultados concernentes ao estudo entoacional da fala de duas variedades do estado do Pará, a cametaense e a mocajubense, resultado de descrição e análise acústica. Uma observação minuciosa das curvas melódicas referentes à F0 da sílaba tônica do último vocábulo das sentenças em escopo, nas três pautas acentuais do português (proparoxítona, paroxítona e oxítona) e nas duas modalidades frasais, declarativa neutra e interrogativa total, denotou distinção entoacional relevante, que ocorre na sílaba pretônica e tônica dos vocábulos analisados. Notou-se, claramente, para a variedade de Cametá a presença de curvas com pouca diferença de inclinação de F0 intersilábica. O comportamento “monotônico” das curvas melódicas de F0 encontrado para Cametá será ponto de investigação para os próximos estudos, posto que a causa desse comportamento, talvez seja a influência de algum substrato linguístico. O parâmetro acústico de Duração denotou maior duração de tempo na fala dos locutores do Cametá, o fenômeno incidiu nos vocábulos correspondentes as três pautas acentuais, já a Intensidade não demonstrou variação significativa. Nesse sentido, estas considerações contribuem para a caracterização prosódica do falar na Amazônia. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 4 Análise e discussão dos resultados Obtiveram-se as seguintes respostas: a) O padrão entoacional encontrado para as frases na modalidade declarativa neutra, falada por locutores da cidade de Cametá não é similar ao encontrado para Mocajuba, pois para Cametá registrou-se pouca variação de movimento de F0 na formação do contorno entoacional, enquanto que os resultados de Mocajuba registraram uma variação significativa intersilábica configurando o movimento ascendente na pretônica e descendente na tônica encontrado por Moraes (2003 apud SILVA e CUNHA, 2012, p. 10) para descrever o português brasileiro; a) O padrão entoacional encontrado para as frases na modalidade declarativa neutra, falada por locutores da cidade de Cametá não é similar ao encontrado para Mocajuba, pois para Cametá registrou-se pouca variação de movimento de F0 na formação do contorno entoacional, enquanto que os resultados de Mocajuba registraram uma variação significativa intersilábica configurando o movimento ascendente na pretônica e descendente na tônica encontrado por Moraes (2003 apud SILVA e CUNHA, 2012, p. 10) para descrever o português brasileiro; b) O padrão entoacional encontrado para as interrogativas da cidade de Cametá não é semelhante ao encontrado para Mocajuba, uma vez que, os movimentos melódicos das curvas de F0 para Cametá registraram altura de F0 uniforme, nas sílabas, alvo de análise, não denotando variação significativa no que concerne a altura melódica, nos três níveis de escolaridade analisados. Por outro lado, Mocajuba registrou uma musicalidade na variação de F0, haja vista que apresentou variação entoacional intersilábica nos vocábulos das três pautas acentuais, principalmente nos níveis médio e superior de escolaridade; c) Os padrões entoacionais encontrados para as interrogativas totais, da variedade de Mocajuba são os mesmos encontrados para a cidade de Brejo (MA) por Silva e Cunha (2012). As autoras descrevem que nas interrogativas há uma subida melódica da F0 na sílaba tônica do acento nuclear, que se estende à postônica final, caracterizando um movimento ascendente nas interrogativas, este movimento foi melhor observado nos resultados das paroxítonas de Mocajuba, nos três níveis de escolaridade; d) As declarativas, referentes a variedade de Mocajuba, assemelham-se, ao padrão entoacinal encontrado por Reis e Cunha (2012) para as declarativas, na região nuclear da sentença, correspondente a variedade de Recife. O movimento é identificado com um tom alto na sílaba pretônica, sofrendo uma descendência nas sílabas tônicas e pós- tônicas. Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. Recebido em 05/10/2019 e aprovado em 22/11/2019. Referências Revista Tabuleiro de Letras (PPGEL, Salvador, online), vol.: 13; n. 3, Especial, dezembro de 2019. ISSN: 2176-5782. 67 Análise entoacional na Amazônia: um estudo comparativo interdialetal das variedades do português falado em Mocajuba e Cametá (PA) COSTA, Maria Sebastiana Silva. Estudo comparativo e perceptual do português falado na Amazônia: a influência da LGA nos dialetos amazônicos. Tese (Doutorado em Letras) – UFPA/ILC/PPGL, Belém, Em andamento. ______. Análise acústica da relação acento versus entoação no português falado em Mocajuba: contribuições para o projeto AMPER Norte. 2015. 150 f. Dissertação (Mestrado em Letras) – UFPA/ILC/CML, Belém, 2015. CRUZ, Regina et al. Formação e Anotação do Corpus do Projeto AMPER Norte. In: Proceedings of GSCP 2012, Belo Horizonte: UFMG, fevereiro de 2012. FOULKES, Paul; SCOBBIE, James; WATT, Dominic. Sociophonetics. In: HARDCASTLE, W.; LAVER, J.; GIBBON, F. (Orgs.). The handbook of Phonetic Sciences. Oxford: Wiley-Blackwell, 2010. REIS, Gizelly, F. M.; CUNHA, Cláudia de Souza. Enunciados assertivos e interrogativos na prosódia maranhense: resultados preliminares. Revista Littera Online, Maranhão, Nº. 5, p. 7-22, 2012. SILVA, Joelma Castelo Branco; CUNHA, Cláudia de Souza. Caracterização prosódica dos falares brasileiros: A oração interrogativa total. Revista do GELNE, Natal, v. 14, p. 59-75, 2012. SORIANO, Larissa Grasiela M. Percepções sociofonéticas do (-R) em São Paulo. 2016. 137 f. Dissertação (Mestrado em Letras). Faculdade de Filosofia, Letras e Ciências Humanas, Universidade de São Paulo, São Paulo. Recebido em 05/10/2019 e aprovado em 22/11/2019. 68
https://openalex.org/W2900054628	https://europepmc.org/articles/pmc6219404?pdf=render	English	null	De novo assembly and annotation of three Leptosphaeria genomes using Oxford Nanopore MinION sequencing	Scientific data	2,018	cc-by	8,797	www.nature.com/scientificdata www.nature.com/scientificdata Received: 5 April 2018 Accepted: 13 July 2018 Published: 6 November 2018 1Genoscope, Institut de Biologie François-Jacob, Commissariat à lʼEnergie Atomique (CEA), Université Paris- Saclay, F-91057 Evry, France. 2UMR BIOGER, INRA, AgroParisTech, Université Paris-Saclay, Avenue Lucien Brétignières, BP 01, F-78850 Thiverval-Grignon, France. 3Génomique Métabolique, Genoscope, Institut de Biologie François Jacob, Commissarait à lʼEnergie Atomique (CEA), CNRS, Université dʼEvry, Université Paris- Saclay, 91057 Evry, France. Correspondence and requests for materials should be addressed to M.-H.B. (email: marie-helene.balesdent@inra.fr) or J.-M.A. (email: jmaury@genoscope.cns.fr) Background & Summary h l g y Fungi have an important ecological and economic role. Many of them are pathogens of animals or plants, especially Leptosphaeria maculans which is an ascomycete phytopathogen of Brassica napus (oilseed rape, canola). Two genomes of Leptosphaeria were already available, the ﬁrst genome sequence of this fungus (JN3) was obtained in the 2000’s using Sanger sequencing1 and composed of 41 scaffolds totaling 43.76 Mb (excluding ambiguous bases). In-depth analysis of this genome showed an unusual compartmenta- lization between AT-rich and GC-equilibrated blocks termed isochores. The location of effector genes in the plastic, AT-rich compartments of the genome was instrumental in establishing the “two-speed fungal genome” paradigm1. According to this paradigm, phytopathogen genomes encompass a plastic compartment enriched in genes involved in niche adaptation. Similar genome organisation was later on found to be a commonality in numerous ﬁlamentous phytopathogens, but its identiﬁcation was slowed down by the use of short-reads sequencing that tended to misrepresent the plastic, Transposable Element (TE)-rich genome in the assemblies. Thus the genome sequencing of Leptosphaeria biglobosa (B3.5), a species related to L. maculans resulted in a poor-quality assembly with complete absence of a putative dispensable genome2. This assembly of B3.5 was composed of 606 scaffolds totaling 29.44 Mb (excluding ambiguous bases). g g Generally, when using short-read sequencing strategies, TE complicate the assembly step and it results in an under estimation of the repetitive content by collapsing similar copies of repetitions. In this study, we generated genome assembly and annotation of three Leptosphaeria isolates. To overcome the limitation of short reads, we generated long reads using the Oxford Nanopore technology (Data Citation 1–3). First, we resequenced the JN3 isolate with the objective of improving the existing assembly in terms of both contiguity and gaps closure (Fig. 1). And then we de novo sequenced and assembled two isolates L. maculans Nz-T4 and L. biglobosa G12-14 (Fig. 1). For each isolate, we generated long and short reads using respectively the MinION device from Oxford Nanopore and the Illumina sequencing technology (Tables 1, 2, 3). Reconciling manually the data from nanopore sequencing, optical and genetic maps allowed the generation of a chromosome-scale assembly for JN3 composed of 19 chromosomes plus the conditionally dispensable chromosome3, with only four missing telomeric extremities. The three genome assemblies were composed of 33, 288 and 156 scaffolds totaling 45.99 Mb, 43.42 Mb and 34.95 Mb for JN3, Nz-T4 and G12-14, respectively (Table 4). Background & Summary h l Furthermore, the gene prediction was supported by integrating conserved proteins and RNA sequencing from Leptosphaeria-infected samples (Fig. 2 and Data Citation 4). It resulted in 13,047, 14,026 and 12,678 predicted protein-coding genes for JN3, Nz-T4 and G12-14, respectively (Table 5 and Fig. 3). Data Descriptor: De novo assembly and annotation of three Leptosphaeria genomes using Oxford Nanopore MinION sequencing (email: jmaury@genoscope.cns.fr) SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Design Type(s) sequence assembly objective • transcription proﬁling by high throughput sequencing design • sequence annotation objective Measurement Type(s) genome assembly • sequence annotation Technology Type(s) whole genome sequencing • RNA sequencing Factor Type(s) Organism Strain • Biospecimen Type • cultivar • culture medium • sampling time Sample Characteristic(s) Leptosphaeria maculans 'brassicae' group • Leptosphaeria biglobosa 'brassicae' group • Leptosphaeria maculans species complex Design Type(s) sequence assembly objective • transcription proﬁling by high throughput sequencing design • sequence annotation objective Measurement Type(s) genome assembly • sequence annotation Technology Type(s) whole genome sequencing • RNA sequencing Factor Type(s) Organism Strain • Biospecimen Type • cultivar • culture medium • sampling time Sample Characteristic(s) Leptosphaeria maculans 'brassicae' group • Leptosphaeria biglobosa 'brassicae' group • Leptosphaeria maculans species complex Data Descriptor: De novo assembly and annotation of three Leptosphaeria genomes using Oxford Nanopore MinION sequencing Received: 5 April 2018 Accepted: 13 July 2018 Published: 6 November 2018 Fabien Dutreux1,2, Corinne Da Silva1, Léo d’Agata1, Arnaud Couloux1, Elise J. Gay2, Benjamin Istace1, Nicolas Lapalu2, Arnaud Lemainque1, Juliette Linglin2, Benjamin Noel1, Patrick Wincker3, Corinne Cruaud1, Thierry Rouxel2, Marie-Hélène Balesdent2 & Jean-Marc Aury1 Leptosphaeria maculans and Leptosphaeria biglobosa are ascomycete phytopathogens of Brassica napus (oilseed rape, canola). Here we report the complete sequence of three Leptosphaeria genomes (L. maculans JN3, L. maculans Nz-T4 and L. biglobosa G12-14). Nz-T4 and G12-14 genome assemblies were generated de novo and the reference JN3 genome assembly was improved using Oxford Nanopore MinION reads. The new assembly of L. biglobosa showed the existence of AT rich regions and pointed to a genome compartmentalization previously unsuspected following Illumina sequencing. Moreover nanopore sequencing allowed us to generate a chromosome-level assembly for the L. maculans reference isolate, JN3. The genome annotation was supported by integrating conserved proteins and RNA sequencing from Leptosphaeria-infected samples. The newly produced high-quality assemblies and annotations of those three Leptosphaeria genomes will allow further studies, notably focused on the tripartite interaction between L. maculans, L. biglobosa and oilseed rape. The discovery of as yet unknown effectors will notably allow progress in B. napus breeding towards L. maculans resistance. 1Genoscope, Institut de Biologie François-Jacob, Commissariat à lʼEnergie Atomique (CEA), Université Paris- Saclay, F-91057 Evry, France. 2UMR BIOGER, INRA, AgroParisTech, Université Paris-Saclay, Avenue Lucien Brétignières, BP 01, F-78850 Thiverval-Grignon, France. 3Génomique Métabolique, Genoscope, Institut de Biologie François Jacob, Commissarait à lʼEnergie Atomique (CEA), CNRS, Université dʼEvry, Université Paris- Saclay, 91057 Evry, France. Correspondence and requests for materials should be addressed to M.-H.B. (email: marie-helene.balesdent@inra.fr) or J.-M.A. Biological material g Two L. maculans brassicae isolates, JN3 (v23.1.3) and Nz-T4 were sequenced here. JN3 is the reference sequenced isolate1 of European origin. Nz-T4 has been used in previous genetic analyses4 and originates from New Zealand. The L. biglobosa brassicae isolate G12-14, also sequenced here, was isolated in 2014 in France (Thiverval-Grignon) as a single-ascospore isolate from naturally infected oilseed rape stubble. G12-14 was chosen as a new L. biglobosa reference isolate since the previously sequenced isolate, B3.52, showed a very low level of aggressiveness when used in cotyledon inoculation tests. In contrast, G12-14 rapidly produced on oilseed rape plants typical and intense symptoms of L. biglobosa brassicae. In addition it was considered as a better representative of the current L. biglobosa populations infecting oilseed rape in France than B3.5. p For RNA sequencing, additional isolates were used, including L. maculans JN2 (v23.1.2), a sister isolate of JN3 with a high level of aggressiveness and the L. biglobosa brassicae reference isolate, B3.52. Plant materials infected with L. maculans and/or L. biglobosa were obtained under various conditions (in the ﬁeld or under controlled conditions) and with three different B. napus varieties. Naturally infected plant material was obtained in Thiverval-Grignon experimental ﬁelds as either infected stem base at the SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Nanopore 2D reads SmartDeNovo Pilon de novo genome assembly PBJelly Scaffolding and gapclosing Illumina reads High quality assembly Polishing G1214 & Nz-T4 JN3 G1214 & Nz-T4 x2 All Nanopore reads JN3 reference genome Chromosome-scale assembly Optical & genetic maps Figure 1. General description of the assembly workﬂow. Amelioration of the existing JN3 assembly and de novo assembly of Nz-T4 and G12-14 isolates. Figure 1. General description of the assembly workﬂow. Amelioration of the existing JN3 assembly and de novo assembly of Nz-T4 and G12-14 isolates. Table 1. Metrics of raw nanopore datasets. Table 1. Metrics of raw nanopore datasets. Biological material Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa G12-14 All Number of reads 1,736,075 (23 ﬂowcells) 883,625 (30 ﬂowcells) 931,376 (19 ﬂowcells) Cumulative size 6,044,436,091 4,813,632,244 3,362,956,224 Estimated coverage 134X 107X 96X Average Size (bp) 3,482 5,448 3,611 Longest read (bp) 1,612,597 1,769,119 949,732 N50 (bp) 6,441 7,197 7,115 # reads > 10Kb 72,482 76,176 61,856 1D Number of reads 1,278,253 443,063 482,374 Cumulative size 4,402,377,873 2,272,788,773 1,693,737,429 Estimated coverage 98X 51X 48X Average Size (bp) 3,444 5,130 3,511 Longest read (bp) 1,612,597 1,769,119 949,732 N50 (bp) 6,431 7,497 7,250 # reads > 10Kb 52,999 42,132 33,148 2D Number of reads 457,822 440,562 449,002 Cumulative size 1,642,058,218 2,540,843,471 1,669,218,795 Estimated coverage 36X 56X 48X Average Size (bp) 3,587 5,767 3,718 Longest read (bp) 237,404 301,948 84,061 N50 (bp) 6,469 7,008 7,005 # reads > 10Kb 19,483 34,044 28,708 SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa G12-14 Number of reads 36,421,608 19,106,872 22,108,636 Cumulative size 8,089,956,155 4,399,409,082 4,198,089,452 Estimated coverage 180 X 98 X 120 X Size (bp) 2×251 # HiSeq2500 2×251 # MiSeq 2×251# MiSeq Table 2. Metrics of illumina datasets. status Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa G12-14 complete dataset 2D dataset 2D dataset Number of reads 1,736,075 440,562 449,002 Cumulative size 6,044,436,091 2,540,843,471 1,669,218,795 Estimated coverage 134X 56X 48X Average Size (bp) 3,482 5,767 3,718 Longest read (bp) 1,612,597 301,948 84,061 N50 (bp) 6,441 7,008 7,005 # reads > 10Kb 72,482 34,044 28,708 status Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa G12-14 complete dataset 2D dataset 2D dataset Number of reads 1,736,075 440,562 449,002 Cumulative size 6,044,436,091 2,540,843,471 1,669,218,795 Estimated coverage 134X 56X 48X Average Size (bp) 3,482 5,767 3,718 Longest read (bp) 1,612,597 301,948 84,061 N50 (bp) 6,441 7,008 7,005 # reads > 10Kb 72,482 34,044 28,708 Table 3. Metrics of nanopore datasets used for genome assemblies. Reference Leptosphaeria maculans JN3 Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa B3.5 Leptosphaeria biglobosa G12-14 Rouxel et al.1 This study This study Grandaubert et al.2 This study # sequences 41 33 288 606 156 Cumulative size 44,892,605 45,986,477 43,426,637 31,788,051 34,950,111 N50 1,769,547 2,437,616 383,462 779,070 462,395 N90 1,020,521 1,391,278 64,152 125,916 123,107 L50 10 8 38 14 22 L90 22 18 150 49 77 # of N’s 1,128,152 (2.51%) 489,445 (1.06%) 0 (0%) 2,343,201 (7.37%) 0 (0%) % GC 45.24% 45.22% 45.69% 51.39% 49.13% Table 4. Biological material Metrics of the existing and new assemblies. p g end of the growing season, sampled one week before harvest (variety Darmor-Bzh) or as whole stem residues sampled two weeks after harvest (variety Alpaga). Under controlled conditions, two varieties were used, Darmor-Bzh displaying a high level of ﬁeld resistance to L. maculans, and Bristol, with a high level of susceptibility. Cotyledons and stems were inoculated as previously described5 and inoculated material was sampled at different time points after inoculation. For petiole inoculations, the petiole was cut under the leaf blade. Ten μL of inoculum (107 pycnidiospores mL−1) were placed over the wound. Inoculated plants were incubated in darkness for 48 h at room temperature under saturated humidity, and then transferred to a growth chamber at 16 °C (night) and 24 °C (day), with a 16 h photoperiod. Infected petioles were sampled after 7 and 14 days post infection (dpi). end of the growing season, sampled one week before harvest (variety Darmor-Bzh) or as whole stem residues sampled two weeks after harvest (variety Alpaga). Under controlled conditions, two varieties were used, Darmor-Bzh displaying a high level of ﬁeld resistance to L. maculans, and Bristol, with a high level of susceptibility. Cotyledons and stems were inoculated as previously described5 and inoculated material was sampled at different time points after inoculation. For petiole inoculations, the petiole was cut under the leaf blade. Ten μL of inoculum (107 pycnidiospores mL−1) were placed over the wound. Inoculated plants were incubated in darkness for 48 h at room temperature under saturated humidity, and then transferred to a growth chamber at 16 °C (night) and 24 °C (day), with a 16 h photoperiod. Infected petioles were sampled after 7 and 14 days post infection (dpi). RNA/DNA extraction h f d l d www.nature.com/sdata/ Reference Leptosphaeria maculans JN3 Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa B3.5 Leptosphaeria biglobosa G12-14 Rouxel et al.1 This study This study Grandaubert et al.2 This study # genes 12,611 13,047 14,026 11,390 12,678 # mono-exonic genes 2,931 5,204 5,898 2,726 4,630 Gene length (avg:med) 1,592:1,278 1,652:1,341 1,507:1,211 1,501:1,217 1,679:1,337 # exons per gene (avg:med) 2.94:2 2.28:2 2.20:2 2.67:2 2.35:2 CDS length (avg:med) 1,392:1,110 1,177:891 1,107:795 1,307:1,065 1,208:954 # introns 24,475 16,700 16,887 18,998 17,076 introns length (avg:med) 103:63 127:57 98:57 116:57 151:56 Coding fraction 38.9% 33.4% 35.8% 46.8% 43.8% BUSCO (euk) 89% 98% 96% 93% 96% BUSCO (fungi) 84% 96% 93% 90% 95% Table 5. Metrics of the existing and new gene predictions. Repeats identification Proteins Combiner RNAseq Quality control Genome visualization RNA-Seq samples Reads are assembled and resulting contigs are aligned on genome assembly Oases, blat, est2genome Genome masking Low complexity, known repeats… RepeatMasker, TRF, dust, RepeatScout Related proteomes Blat, blast, GeneWise Gene Prediction Gmove Quality evaluation of gene predictions Exon/inton structures and alignment with Uniprot proteins http://www.genoscope.cns.fr/leptolife Figure 2. General description of the gene prediction workﬂow. Repeats identification Proteins Combiner RNAseq Quality control Genome visualization RNA-Seq samples Reads are assembled and resulting contigs are aligned on genome assembly Oases, blat, est2genome Genome masking Low complexity, known repeats… RepeatMasker, TRF, dust, RepeatScout Related proteomes Blat, blast, GeneWise Gene Prediction Gmove Quality evaluation of gene predictions Exon/inton structures and alignment with Uniprot proteins http://www.genoscope.cns.fr/leptolife Quality evaluation of gene predictions Exon/inton structures and alignment with Uniprot proteins Figure 2. General description of the gene prediction workﬂow. Reference Leptosphaeria maculans JN3 Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa B3.5 Leptosphaeria biglobosa G12-14 Rouxel et al.1 This study This study Grandaubert et al.2 This study # genes 12,611 13,047 14,026 11,390 12,678 # mono-exonic genes 2,931 5,204 5,898 2,726 4,630 Gene length (avg:med) 1,592:1,278 1,652:1,341 1,507:1,211 1,501:1,217 1,679:1,337 # exons per gene (avg:med) 2.94:2 2.28:2 2.20:2 2.67:2 2.35:2 CDS length (avg:med) 1,392:1,110 1,177:891 1,107:795 1,307:1,065 1,208:954 # introns 24,475 16,700 16,887 18,998 17,076 introns length (avg:med) 103:63 127:57 98:57 116:57 151:56 Coding fraction 38.9% 33.4% 35.8% 46.8% 43.8% BUSCO (euk) 89% 98% 96% 93% 96% BUSCO (fungi) 84% 96% 93% 90% 95% Figure 2. General description of the gene prediction workﬂow. Table 5. Metrics of the existing and new gene predictions. Table 5. Metrics of the existing and new gene predictions. instructions. RNA/DNA extraction h f d l d Fresh infected cotyledons or freeze-dried cultures of fungal mycelia were ground using a Retsch MM300 mixer mill in Eppendorf tubes, with one tungsten carbide bead per tube, for 45 s with 30 oscillations per second. Petioles were ground with a pestle and mortar. Stem base and stem residues were ground with a Retsch MM300 mixer mill using Zirconium oxide jars and beads, for 40 s with 30 oscillations per second. For RNA extraction, all grinding material was frozen with liquid nitrogen prior grinding. RNA extraction was then performed using the Trizol reagent (Invitrogen, Cergy Pontoise, France) as previously described6. DNA was extracted from freeze-dried material using the plant DNeasy mini-kit or the Wizard® Genomic DNA Puriﬁcation Kit (Promega, Madison, WI, USA) according to the manufacturers’ SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ instructions. Input DNA for the nanopore sequencing, based on long DNA molecules, was extracted from agarose plugs of concentrated pycnidiospores (>1.109 spore ml−1) partially digested1,3. Brieﬂy, fungal 3 Reference Leptosphaeria maculans JN3 Leptosphaeria maculans JN3 Leptosphaeria maculans Nz-T4 Leptosphaeria biglobosa B3.5 Leptosphaeria biglobosa G12-14 Rouxel et al.1 This study This study Grandaubert et al.2 This study # genes 12,611 13,047 14,026 11,390 12,678 # mono-exonic genes 2,931 5,204 5,898 2,726 4,630 Gene length (avg:med) 1,592:1,278 1,652:1,341 1,507:1,211 1,501:1,217 1,679:1,337 # exons per gene (avg:med) 2.94:2 2.28:2 2.20:2 2.67:2 2.35:2 CDS length (avg:med) 1,392:1,110 1,177:891 1,107:795 1,307:1,065 1,208:954 # introns 24,475 16,700 16,887 18,998 17,076 introns length (avg:med) 103:63 127:57 98:57 116:57 151:56 Coding fraction 38.9% 33.4% 35.8% 46.8% 43.8% BUSCO (euk) 89% 98% 96% 93% 96% BUSCO (fungi) 84% 96% 93% 90% 95% Table 5. Metrics of the existing and new gene predictions. Repeats identification Proteins Combiner RNAseq Quality control Genome visualization RNA-Seq samples Reads are assembled and resulting contigs are aligned on genome assembly Oases, blat, est2genome Genome masking Low complexity, known repeats… RepeatMasker, TRF, dust, RepeatScout Related proteomes Blat, blast, GeneWise Gene Prediction Gmove Quality evaluation of gene predictions Exon/inton structures and alignment with Uniprot proteins http://www.genoscope.cns.fr/leptolife Figure 2. General description of the gene prediction workﬂow. Illumina PCR-Free library preparation and sequencing ( ) d b Illumina PCR Free library preparation and sequencing DNA (4.5 to 6 μg) was sonicated to a 100- to 1500-bp size range using a Covaris E210 sonicator (Covaris, Woburn, MA, USA). Fragments were end-repaired using the NEBNext End Repair Module (New England Biolabs, Ipswich, MA, USA) and 3΄-adenylated with the NEBNext dA-Tailing Module (New England Biolabs). Illumina adapters were then added using the NEBNext Quick Ligation Module (New England Biolabs) and ligation products were puriﬁed with AMPure XP beads (Beckmann Coulter Genomics, Danvers, MA, USA). Libraries were sequenced on an Illumina MiSeq (G12-14 and NzT4 genomes) or a HiSeq 2500 (JN3 genome) instrument (San Diego, CA, USA) using 250 base-length read chemistry in a paired-end mode. RNA/DNA extraction h f d l d Input DNA for the nanopore sequencing, based on long DNA molecules, was extracted from agarose plugs of concentrated pycnidiospores (>1.109 spore ml−1) partially digested1,3. Brieﬂy, fungal pycnidiospores were collected from V8-agar plates3 with sterile distilled water, centrifuged (15 min at 10000 ´ g) and adjusted to 6.109 spore mL1. The spore suspension was mixed with an equal volume of 2.5% low melting agarose (Seaplaque GTG® in TSE; Tris 25 mM, pH 7.5; Sorbitol 1 M, EDTA 25 mM) maintained at 40 °C. This suspension was poured in disposable plug molds (BioRad). After cooling, the resulting plugs were treated for 20 h in 0.5 M EDTA, SDS 10%, 1 mg.mL −1 proteinase K (Sigma) at 50 °C under mild stirring, then rinsed three times with 0.5 M EDTA, 50 °C and stored at 4 °C in 0.5 M EDTA. For a part of Nanopore sequencing runs, DNA was also extracted from freeze-dried mycelium ground in liquid nitrogen: cells have been then lysed using CTAB (Cetyltrimethylammonium bromide) with 2% of PVP (Polyvinylpyrrolidone) and 0.1% of 2-mercaptoéthanol and DNA was puriﬁed following a phenol- chloroform protocol. 5 SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Figure 3. Genome browser database for the three Leptosphaeria isolates. The genome browser is available at http://www.genoscope.cns.fr/leptolife and contains repeats (green track), coverage of genomic reads (black wiggle), gene prediction (blue track), RNA contigs (dark green track) and protein homologies (salmon track). Figure 3. Genome browser database for the three Leptosphaeria isolates. The genome browser is available at http://www.genoscope.cns.fr/leptolife and contains repeats (green track), coverage of genomic reads (black wiggle), gene prediction (blue track), RNA contigs (dark green track) and protein homologies (salmon track). Figure 3. Genome browser database for the three Leptosphaeria isolates. The genome browser is available at http://www.genoscope.cns.fr/leptolife and contains repeats (green track), coverage of genomic reads (black wiggle), gene prediction (blue track), RNA contigs (dark green track) and protein homologies (salmon track). Illumina PCR-Free library preparation and sequencing DNA (4 5 t 6 ) i t d t 100 t 1500 b i Illumina cDNA library preparation and sequencing y p p q g RNA-Seq library preparations were carried out from 1–2 μg total RNA using the TruSeq Stranded mRNA kit (Illumina, San Diego, CA, USA), which allows mRNA strand orientation (sequence reads occur in the same orientation as anti-sense RNA). Brieﬂy, poly(A)+ RNA was selected with oligo(dT) beads, chemically fragmented and converted into single-stranded cDNA using random hexamer priming. Then, the second strand was generated to create double-stranded cDNA. cDNA were then 30-adenylated, and Illumina adapters were added. Ligation products were PCR-ampliﬁed. Each library was sequenced using 101 bp paired end reads chemistry on a HiSeq2000 Illumina sequencer. Nanopore 8-kb and 20-kb libraries preparation p p p MinION sequencing libraries were prepared according to the nanopore Sequencing Kit protocol SQK- MAP006 or the Low Input Expansion Pack protocol for genomic DNA. Brieﬂy, 100 ng to 1.5 μg of genomic DNA was sheared to approximately 8 Kb with g-TUBE (Covaris, Woburn, MA, USA) and SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ cleaned-up using AMPure XP beads (Beckmann Coulter Genomics). For some libraries, DNA fragments were repaired using the NEBNext FFPE Repair Mix (New England Biolabs). After end-reparation and 30- adenylation with the NEBNext® UltraTM II End Repair/dA-Tailing Module (New England Biolabs), sequencing adapters provided by Oxford Nanopore Technologies (Oxford Nanopore Technologies Ltd, UK) were ligated using Blunt/TA Ligase Master Mix (New England Biolabs). Libraries were then enriched using Dynabeads MyOne C1 Streptavidin (ThermoFisher Scientiﬁc, Wilmington, DE, USA). g y y p g Nanopore 20-Kb libraries were prepared according to the same protocol, with the exception that 50 ng to 2.5 μg of genomic DNA was sheared to approximately 20-kb with g-TUBE (Covaris). Genome assembly update of L. maculans JN3 Genome assembly update of L. maculans JN3 With 33 scaffolds, a N50 equal to 2.40 Mb and a cumulative size of 44.8 Mb, the JN3 rearranged reference genome was already contiguous enough for our analyses. However, it contained gaps that represented 1.1 Mb in total. In addition, both the optical map2 and the high-density genetic map indicated that nine mis-assembled super contigs had to be either splitted or fused. The reference genome was scaffolded and gap-ﬁlled using PBJelly14 with the entire nanopore dataset as input (Table 3). As a result we obtained 33 scaffolds with a cumulative size of 45.8 Mb, the N50 increased to 2.43 Mb and we decreased by half the number of N’s (570 kb). 15 As the R7.3 nanopore reads were prone to errors, particularly in the homopolymeric regions15, we polished the consensus of the gap closed assembly by aligning 150X of Illumina paired-end 2 × 250 bp reads with BWA mem12 and giving the resulting alignment ﬁle to Pilon13. The ﬁnal assembly had a cumulative size of 45.99 Mb with 1% of unknown bases (Table 4). MinION sequencing and basecalling Read event data generated by MinKNOW control software (version 0.50.2.15 to 0.51.3.40) were base- called using the Metrichor software (version 2.34.3 to 2.39.3). The data generated (pores metrics, sequencing, and base-calling data) by MinION software were stored and organized using a Hierarchical Data Format. Three types of reads were obtained: template, complement, and two-directions (2D). The template and complement reads correspond to sequencing of the two DNA strands. Metrichor combines template and complement reads to produce a consensus (2D) sequence7. FASTA reads were extracted from MinION Hierarchical Data Format ﬁles using poretools8. Pass and fail reads were both considered as 1D reads. This dataset is described in Table 1 and (Genomic datasets, Data Citation 5). MinION ﬂow cell preparation and sample loading Each library was mixed with the fuel mix and the running buffer according to the SQK-MAP006 or the Low Input Expansion Pack protocols. The sequencing mix was then added to the R7.3 ﬂowcell for a 48- hour run. Several loading schedules were tested, but the main one used for the MAP006 libraries has been the following: a ﬁrst loading with 8 μl of the library, then three reloading after 5, 24 and 29 h of run with respectively 4 μl, 8 μl and 4 μl of the DNA library. Regarding the low-input libraries, the main schedule used was a ﬁrst loading with 10 μl of the DNA library and a reloading after 24 hours of run time with 10 μl of the DNA library. Genetic map A genetic map was built using 93 progeny of the JN2 ´ Nz-T4 in vitro cross9. A total of 150 polymorphic micro- and minisatellite markers, three biological markers (Mat locus, AvrLm3 and AvrLm7) and eight markers designed at the junction between two repeated elements10 were ﬁrst used to generate an unsaturated genetic map using Mapmaker 3.0, with a LOD score of 3 and a maximum genetic distance of 20 cM. The 93 progeny isolates and the two parental isolates were also sequenced (2 × 100 bp illumina sequencing; 48 samples per line; Beckman Coulter Genomics). SNP calling between parental isolates was done as described in the study11 following mapping of the reads on the JN3 reference genome. A total of 20,103 reliable SNPs were detected and assigned to one of the two parental isolate in the progeny. These SNPs were converted into 2,104 Bins. From all these markers, a saturated map comprising 20 linkage groups was constructed using MSTmap online. Long reads-based de novo assembly of L. biglobosa G12-14 and L. maculans Nz-T4 R7.3 nanopore 2D reads (Table 3) were assembled into contigs using SmartDeNovo (https://github.com ruanjue/smartdenovo), with a k-mer size parameter set to 21. Eventually, the consensus of the assemb Long reads-based de novo assembly of L. biglobosa G12-14 and L. maculans Nz-T4 R7 3 2D d (T bl 3) bl d i t ti i S tD N (htt // ith b g y g R7.3 nanopore 2D reads (Table 3) were assembled into contigs using SmartDeNovo (https://github.com/ ruanjue/smartdenovo), with a k-mer size parameter set to 21. Eventually, the consensus of the assembly was polished by aligning the Illumina paired-end 2 × 250 bp reads (Table 2) with BWA mem12 and using the resulting alignment ﬁle as input for Pilon13. We iteratively polished twice the assembly. g g p y p y L. biglobosa G12-14 ﬁnal assembly had a cumulative size of 34.9 Mb and a N50 equal to 462 kb. L. maculans Nz-T4 ﬁnal assembly had a cumulative size of 43.4 Mb and a N50 equal to 383 kb. Further usual metrics regarding those assemblies are provided in Table 4. Genome annotation The gene prediction of the three Leptosphaeria genomes was done by combining gene models information computed from transcript and protein alignments (Fig. 2). SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ First, repeated sequences in the reference genomes were masked using multiple tools. Tandem repeats were identiﬁed and masked using Tandem Repeats Finder16. Low complexity regions were identiﬁed and masked using Dust17. Interspersed repeats and other low complexity sequences were masked using RepeatMasker (http://www.repeatmasker.org). Furthermore 121 known transposable elements (identiﬁed in the L. maculans-L. biglobosa species complex in a previous study2) were also masked using RepeatMasker. p The Alternaria alternata, Cochliobolus heterostrophus, Pyrenochaeta sp., Pyrenophora tritici-repentis and Zymoseptoria tritici proteomes included in UniProt (version available as of 07 September 2016) were used to detect conserved proteins in L. maculans JN3, L. maculans Nz-T4 and L. biglobosa G12-14. L. maculans JN3 and swissprot proteins (release available as of 30 November 2016) were also used as a resource to detect conserved proteins in L. maculans Nz-T4 and L. biglobosa G12-14. Those proteins were ﬁrst aligned to L. maculans JN3, L. maculans Nz-T4 and L. biglobosa G12-14 genomes assemblies using BLAT18. BLAT results were then ﬁltered as follows: for each protein, the best match (based on the BLAT score) and other matches with a score > = 90% of the score of the best match were retained. Genewise19 was then used to reﬁne matches and identify exon-intron boundaries and alignment were ﬁltered out if less than 50% of the protein length was aligned. g p g g RNAseq reads from 23 samples were assembled using Velvet20 1.2.07 and Oases21 0.2.08, using a k-mer size of 63 bp. The results of this assembly for each sample are summarized in (Transcriptomic datasets, Data Citation 5). Reads were mapped back to the contigs with BWA-mem12 and the consistent paired-end reads were selected. Chimeric contigs were identiﬁed and splitted (uncovered regions) based on coverage information from consistent paired-end reads. Moreover, open reading frames (ORF) and domains were searched using respectively TransDecoder22 and CDDsearch23. We only allowed breaks outside ORF and domains. At the end, the read strand information was used to correctly orient the RNA- Seq contigs. In a second step, RNAseq contigs were aligned to the relevant genome assembly (L. maculans JN3, L. maculans Nz-T4, L. biglobosa G12-14) using BLAT. Technical Validation DNA and RNA sample quality p q y DNA quality was assessed using 1% agarose gel. RNA integrity was assessed using an Agilent 2 Bioanalyzer electrophoresis system (Agilent, Santa Clara, CA, USA). Genome annotation The best matches (based on BLAT score) for each contig and other matches with a score > = 90% of the score of the best match were selected. Then, Est2genome24 was used to reﬁne the alignments and we kept alignments with an identity percent > = 95% and at least 80% of the RNA-Seq contig that can be aligned (Transcriptomic datasets, Data Citation 5). 25 Gene predictions from mRNA and proteomes were given as input to the GMOVE combiner25 to build the gene models. This tool considers biological data such as RNA-seq from the organisms of interest and proteome from homologous species. Proteins give information about the frame of the coding sequence (CDS) and RNA-seq helps to ﬁnd splicing sites with more accurate alignments. This tool is designed for all eukaryotes and de novo studies with no need of calibration pre-processing. After gene models were produced by GMOVE, a ﬁnal ﬁlter was applied to remove artefactual untranslated regions (UTRs). UTRs were trimmed if it overlapped CDS in predicted gene models for more than 10 bases. Moreover, UTRs overlapping other UTRs for more than 10 bases were also trimmed to avoid large and potentially false UTRs. Final metrics of the gene prediction are available in Table 5 and Fig. 3 is a screenshot of the genome browser. Data Records The authors declare that all data reported herein are fully and freely available from the date of publication. Genomic data have been deposited in the ENA repository (Genomic datasets, Data Citation 5): L. maculans JN3 (Data Citation 1), L. maculans Nz-T4 (Data Citation 2) and L. biglobosa G12-14 (Data Citation 3). RNA data used for gene prediction have been deposited in the ENA repository (Data Citation 4 and Transcriptomic datasets, Data Citation 5). Moreover, a dedicated website brings together all the information: accession numbers, assembly and annotation ﬁles as well as a genome browser (Fig. 3): http:// www.genoscope.cns.fr/leptolife. Code availability y All the tools used for this study are cited in the method section and they were used with default settings unless options were speciﬁed. The in-house tools to clean illumina data are available from the following website: http://www.genoscope.cns.fr/fastxtend/. The integration of resources in the gene prediction workﬂow was achieved using GMOVE: http://www.genoscope.cns.fr/gmove/. Illumina libraries Ready-to-sequence Illumina libraries were quantiﬁed by qPCR using the KAPA Library Quantiﬁcation Kit for Illumina Libraries (KapaBiosystems, Wilmington, MA, USA), and libraries proﬁles evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Reference LmJN3∗ LmJN3 LmNz-T4 LbB3.5 LbG12-14 Rouxel et al.1 This study This study Grandaubert et al.2 This study GC blocks # blocks 399 534 587 389 318 % assembly 64 63.1 66.6 95.1 84.5 mean size (kb) 70.4 54.4 49.2 77.7 92.9 stdev (kb) — 63.1 61.9 226 157 min (kb) 1 1 1 1 1 max (kb) 500 431 349 1708 1180 # genes — 12,892 13,790 11,361 12,603 AT blocks # blocks 413 564 779 324 335 % assembly 36 36.9 33.4 4.87 15.5 mean size (kb) 38.6 30.1 18.6 4.8 16.2 stdev (kb) — 48.1 23.5 10.7 17.7 min (kb) 1 1 1 1 1 max (kb) 320 319 245 120 142 # genes — 233 236 29 75 Table 6. Genome compartmentalization of old and new assemblies. ∗Results from Grandaubert et al. AT rich regions were manually curated. Table 6. Genome compartmentalization of old and new assemblies. ∗Results from Grandaubert et al. AT rich regions were manually curated. Impact of the gap closing procedure on the JN3 assembly p g p g p y The JN3 assembly was gap closed using PBJelly and we detected several duplicated regions of several Kb in this gap-closed assembly that were not present in the previous assembly. We ﬁrst detected tandemly duplicated genes using Blast and revealed 28 candidate regions totaling 150 kb in length with a median size of 5.5 kb and covering 63 genes. Further investigations of these regions using genomic data coverage revealed that they were artefactual duplicates created by PBJelly. We mapped both Illumina and nanopore reads onto the gap-closed assembly and highlighted genomic regions without illumina and nanopore reads coverage. This method revealed 30 regions with a cumulative size of 185 kb and a median size of 5.1 kb spanning a total of 43 genes. Finally, after a careful expertise using the genome browser (Fig. 3), the genomic regions that presented duplicated genes and no proper read support were masked and overlapping genes were ﬁltered out. In total, we masked 41 regions with a cumulative size of 264 kb and a median size of 5.5 kb that spanned 81 genes (Duplicated genomic regions, Data Citation 5). Illumina reads processing and quality ﬁltering After the Illumina sequencing, an in-house quality control process was applied to the reads that passed the Illumina quality ﬁlters. The ﬁrst step discards low-quality nucleotides (Q o 20) from both ends of the reads. Next, Illumina sequencing adapters and primer sequences were removed from the reads. Then, reads shorter than 30 nucleotides after trimming were discarded. These trimming and removal steps were achieved using in-house-designed software based on the FastX package. The last step identiﬁes and discards read pairs that mapped to the phage phiX genome, using SOAP26 and the phiX reference sequence (GenBank: NC_001422.1). This processing resulted in high-quality data and improvement of the subsequent analyses. This dataset is described in Table 2 and (Genomic datasets, Data Citation 5). A speciﬁc ﬁlter aiming to remove ribosomal reads was applied to data generated from RNA samples. The reads and their mates that mapped onto a ribosomal sequences database were ﬁltered using SortMeRNA27. It contains different rRNA databases and we used it to split the data into two ﬁles: rRNA reads in a ﬁle (ribo_clean) and other reads in another ﬁle (noribo_clean). This dataset is described in (Transcriptomic datasets, Data Citation 5). 1. Rouxel, T. et al. Effector diversiﬁcation within compartments of the Leptosphaeria maculans genome affected by Repeat-Induced Point mutations. Nature communications 2, 202, doi:10.1038/ncomms1189 (2011). Contamination detection In order to detect possible contamination in our assemblies, we ﬁrst ran the metagene28 gene predictor on NzT4 and G12-14 contigs and on JN3 scaffolds. Then, we aligned all predicted proteins against the NR database using blastp. Hits were then ﬁltered by their e-value to only conserve the best hit with an e-value of at least 10 −10. Finally, a contig or scaffold was ﬂagged as non-contaminant if at least 50% of its predicted proteins were assigned as fungi. Following this procedure, we couldn’t detect any contaminant sequences in JN3 and G12-14 assemblies. We detected 13 small contigs that were not assigned as fungi in the NzT4 assembly, but further investigations (blat against the JN3 assembly and blastx using the NCBI blast server) revealed that they were not contaminants. SCIENTIFIC DATA \| 5:180235 \| DOI: 10.1038/sdata.2018.235 www.nature.com/sdata/ Comparison with existing assemblies p g An AT/GC region segmentation was performed on all assemblies with OcculterCut29 to detect potential genome compartmentalization as described in the following study1. L. maculans JN3 and Nz-T4 isolates showed similar genome architecture with a third of their assembly dispatched into AT blocks of 30.1 kb and 18.6 kb on average, respectively. The number of genes present within the AT-blocks was also steady, and ranged between 233 and 236. In contrast, there was a drastic modiﬁcation in genome architecture and genome size between L. biglobosa B3.5, sequenced using only Illumina, and G12-14 sequenced here. The two assemblies differed by more than 10% of their genome content. This difference corresponded to 3 Mb missing in B3.5 and recovered in G12-14 thanks to the nanopore long reads (Table 6). The difference mostly comprised of AT-blocks similar to those found in L. maculans and amounting to 15.5% of the G12-14 assembly vs. only 4.9% in the B3.5 assembly (Table 6). The increase in AT-block content is consistent with the lowering of the average GC content of the L. biglobosa genome in the new assembly (Table 4). p g An AT/GC region segmentation was performed on all assemblies with OcculterCut29 to detect potential genome compartmentalization as described in the following study1. L. maculans JN3 and Nz-T4 isolates showed similar genome architecture with a third of their assembly dispatched into AT blocks of 30.1 kb and 18.6 kb on average, respectively. The number of genes present within the AT-blocks was also steady, and ranged between 233 and 236. Contamination detection In contrast, there was a drastic modiﬁcation in genome architecture and genome size between L. biglobosa B3.5, sequenced using only Illumina, and G12-14 sequenced here. The two assemblies differed by more than 10% of their genome content. This difference corresponded to 3 Mb missing in B3.5 and recovered in G12-14 thanks to the nanopore long reads (Table 6). The difference mostly comprised of AT-blocks similar to those found in L. maculans and amounting to 15.5% of the G12-14 assembly vs. only 4.9% in the B3.5 assembly (Table 6). The increase in AT-block content is consistent with the lowering of the average GC content of the L. biglobosa genome in the new assembly (Table 4). 2. Grandaubert, J. et al. Transposable element-assisted evolution and adaptation to host plant within the Leptosphaeria maculans Leptosphaeria biglobosa species complex of fungal pathogens. BMC genomics 15, 891, doi:10.1186/1471-2164-15-891 (2014). 3. Leclair, S., Ansan-Melayah, D., Rouxel, T. & Balesdent, M. Meiotic behaviour of the minichromosome in the phytopathogenic ascomycete Leptosphaeria maculans. Current genetics 30, 541–548 (1996). Assessment of gene prediction Assessment of gene prediction First, protein sequences of each genome (L. maculans JN3, L. maculans Nz-T4 and L. biglobosa G12-14) were obtained by translating gene CDS and aligned to protein databases (uniprot_sprot and uniprot_trembl 2016_11 release however L. maculans matches were excluded to prevent the introduction of known biases from the previous annotation effort) using blastp. Gene models for which proteins aligned with an e-value o 10 −10 or with a CDS size > = 200 nt were retained. 30 Moreover gene completion for the newly generated annotations was evaluated using BUSCO30 v1.1b1 fungal and eukaryotic conserved genes databases. The relevance in the increase in gene number between the “old” and “new” gene prediction reﬂects a higher accuracy of the “new” annotation as evidenced by careful examination of genes encoding effectors. Effector genes encode for small (less than 300 amino- acids) secreted proteins (SSPs). They are usually poorly predicted de novo and may lack transcriptomic support due to expression in certain stages of parasitism only. The data obtained here drastically increased the number of predicted SSPs in both species (1,082 for L. maculans vs. 651 in the “old” annotation, 875 for L. biglobosa vs. 665 in the previous annotation) and drastically improved the gene models (data not shown). Also this new annotation allowed the identiﬁcation of 75 genes hosted within the AT-block of the genome of L. biglobosa, previously unidentiﬁed (Table 6). References A reference bacterial genome dataset generated on the MinION portable single-molecule nanopore sequencer. GigaScience 3, 22, doi:10.1186/2047-217X-3-22 (2014). p q g ( ) 8. Loman, N. J. & Quinlan, A. R. Poretools: a toolkit for analyzing nanopore sequence data. Bioinformatics 30, d i /bi i f i /b ( ) p q g ( ) 8. Loman, N. J. & Quinlan, A. R. Poretools: a toolkit for analyzing nanopore sequence data. 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https://openalex.org/W4290038689	https://pure.rug.nl/ws/files/622496599/1_s2.0_S0267364922000693_main.pdf	English	null	Legal and technical questions of file system reverse engineering	Computer law and security report/Computer law & security report	2,022	cc-by	12,566	University of Groningen Legal and technical questions of file system reverse engineering Stoykova, Radina; Nordvik, Rune; Ahmed, Munnazzar; Franke, Katrin; Axelsson, Stefan; Toolan, Fergus Published in: Computer Law & Security Review DOI: 10.1016/j.clsr.2022.105725 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2022 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Stoykova, R., Nordvik, R., Ahmed, M., Franke, K., Axelsson, S., & Toolan, F. (2022). Legal and technical questions of file system reverse engineering. Computer Law & Security Review, 46, Article 105725. https://doi.org/10.1016/j.clsr.2022.105725 University of Groningen Legal and technical questions of file system reverse engineering Stoykova, Radina; Nordvik, Rune; Ahmed, Munnazzar; Franke, Katrin; Axelsson, Stefan; Toolan, Fergus Published in: Computer Law & Security Review DOI: 10.1016/j.clsr.2022.105725 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2022 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Stoykova, R., Nordvik, R., Ahmed, M., Franke, K., Axelsson, S., & Toolan, F. (2022). Legal and technical questions of file system reverse engineering. Computer Law & Security Review, 46, Article 105725. https://doi.org/10.1016/j.clsr.2022.105725 University of Groningen University of Groningen Legal and technical questions of file system reverse engineering DOI: 10.1016/j.clsr.2022.105725 DOI: 10.1016/j.clsr.2022.105725 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Stoykova, R., Nordvik, R., Ahmed, M., Franke, K., Axelsson, S., & Toolan, F. (2022). Legal and technical questions of file system reverse engineering. Computer Law & Security Review, 46, Article 105725. https://doi.org/10.1016/j.clsr.2022.105725 Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Radina Stoykova a , b , ∗, Rune Nordvik a , c , Munnazzar Ahmed , Katrin Franke Stefan Axelsson a , d , Fergus Toolan c a Norwegian University of Science and Technology, Gjøvik Norway b University of Groningen Faculty of law, The Netherlands c Norwegian Police University College, Oslo, Norway d Stockholm University, Sweden a Norwegian University of Science and Technology, Gjøvik Norway b University of Groningen Faculty of law, The Netherlands c Norwegian Police University College, Oslo, Norway d Stockholm University, Sweden Copyright strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of th or copyright holder(s), unless the work is under an open content license (like Creative Commons). The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license. 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For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 24-10-2024 computer law & security review 46 (2022) 105725 ∗Corresponding author. E-mail addresses: adi.stoykova@gmail.com (R. Stoykova), rune.nordvik@phs.no (R. Nordvik), munnazzar@gmail.com (M. Ahmed) katrin.franke@ntnu.no (K. Franke), stefan.axelsson@ntnu.no (S. Axelsson), Fergus.Thomas.Toolan@phs.no (F. Toolan). closed source. Every update or improvement by the FS ven- https://doi.org/10.1016/j.clsr.2022.105725 0267-3649/© 2022 Radina Stoykova, Rune Nordvik, Munnazzar Ahmed, Katrin Franke, Stefan Axelsson, Fergus Toolan. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ ) katrin.franke@ntnu.no (K. Franke), stefan.axelsson@ntnu.no (S. Axelsson), Fergus.Thomas.Toolan@phs.no (F. Toolan). https://doi.org/10.1016/j.clsr.2022.105725 0267-3649/© 2022 Radina Stoykova, Rune Nordvik, Munnazzar Ahmed, Katrin Franke, Stefan Axelsson, Fergus Toolan. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ ) a b s t r a c t Keywords: Digital Forensics Reverse Engineering Criminal procedure Intellectual Property Government Vulnerability Disclosure Reverse engineering of file systems is indispensable for tool testing, accurate evidence ac- quisition, and correct interpretation of data structures by law enforcement in criminal inves- tigations. This position paper examines emerging techno-legal challenges from the practice of reverse engineering for law enforcement purposes. We demonstrate that this new context creates uncertainties about the legality of tools and methods used for evidence acquisition and the compliance of law enforcement with obligations to protect intellectual property and confidential information. Further identified are gaps between legal provisions and practice related to disclosure and peer-review of sensitive digital forensic methodology, trade se- crets in investigations, and governmental vulnerability disclosure. It is demonstrated that reverse engineering of file systems is insufficiently addressed by legislators, which results in a lack of file system interpretation and validation information for law enforcement and their dependence on tools. Outlined are recommendations for further developments of dig- ital forensic regulation. © 2022 Radina Stoykova, Rune Nordvik, Munnazzar Ahmed, Katrin Franke, Stefan Axelsson, Fergus Toolan. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ ) © 2022 Radina Stoykova, Rune Nordvik, Munnazzar Ahmed, Katrin Franke, Stefan Axelsson, Fergus Toolan. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ ) facilitating or furthering the reconstruction of events found to be criminal“ [23] . Ergo, sufficient understanding of those digi- tal sources and their underlying functionalities are crucial to capture, organize, and further interpret in order to derive in- ferences of user activities and digital evidence. File systems (FS) are one of the richest sources of user‘s activity, poten- tially keeping track of every file created, modified, copied or deleted during the entire life span of the device. Many file sys- tems are proprietary, patented or a combination of open and closed source. Every update or improvement by the FS ven- Introduction p // g/ /j 0267-3649/© 2022 Radina Stoykova, Rune Nordvik, Munnazzar Ahmed, Katrin Franke, Stefan Axelsson, Fergus Toolan. Published by Elsevier Ltd. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ ) ∗Corresponding author. Legal and technical questions of file system reverse engineering Radina Stoykova a , b , ∗, Rune Nordvik a , c , Munnazzar Ahmed , Katrin Franke a , Stefan Axelsson a , d , Fergus Toolan c Corresponding author. E-mail addresses: adi.stoykova@gmail.com (R. Stoykova), rune.nordvik@phs.no (R. Nordvik), munnazzar@gmail.com (M. Ahmed), katrin.franke@ntnu.no (K. Franke), stefan.axelsson@ntnu.no (S. Axelsson), Fergus.Thomas.Toolan@phs.no (F. Toolan). 4 The source code is not available in closed source file system, and it is arguable if assessing original source code can be consid- ered reverse engineering Introduction 1. In the context of criminal investigation digital forensics is de- fined as “the use of scientifically derived and proven methods towards the preservation, collection, validation, identification, analysis, interpretation, documentation and presentation of digital evidence derived from digital sources for the purpose of computer law & security review 46 (2022) 105725 2 the other hand, they are very valuable for both criminals and law enforcement, because file systems contain records with timestamps of files created or modified on a computer during its life span [14] , thus they are a silent witness of most digi- tal activities. Many vendors‘ efforts are dedicated to ensuring robust security of this data. As already observed, when digi- tal investigators are dealing with new, unknown data struc- tures in the file system, they need to reverse engineer or by- pass security settings in order to gain access to the file system and to understand its structures. A known technique in mo- bile forensics is the use of chip off, but it is obsolete due to encryption [35] , and therefore other techniques need to be ex- ploited. This may expose copyrighted data or security flaws in systems. dor could result in inaccuracy of the digital forensic (DF) tools and an inability to interpret the FS structures by both tools and examiners. Therefore, reverse engineering (RE) is neces- sary for law enforcement agencies (LEA) to understand new FS systems and / or changes in existing data structures and also to determine tool limitations. While most of the efforts in reverse engineering are focused on code research, this pa- per examines the importance of reverse data engineering for data structures and organization analysis within file systems. This is an indispensable part of the forensic examination of file systems for evidence acquisition [43] . Reverse engineering of closed source file systems is nec- essary because the vendor documentation is either missing, treated as a trade secret, or is not sufficient for law enforce- ment needs. In a digital forensic context, reverse engineering includes three main methods of analysis black-box testing, static-code analysis, and runtime analysis [1] . Commonly used are dis- assemblers, debuggers, file formats, system architectures, de- compilers and low level code [1, p. 267] [4] 1.1. Research questions • Why LEAs need to perform Reverse Engineering and what are the current and future challenges of reverse engineer- ing? Black-box testing is based on sending known data or ac- tions to a system (or part thereof), having the system perform its (perhaps undocumented) function, and then assessing the outcome by viewing the results [1, p. 267] . When performing file system reverse engineering we attempt to understand un- documented metadata structures. For example, creating a file is the input action (the known data), the file system driver uses hardware/software (the black box) to create the file, and the relevant outputs are the metadata structures and the file content as it is saved in the file system. File system metadata is any information that describes the files and directories in the file system [11] . Examples of metadata are file size, owner, timestamps, the file content on-disk location, etc. • How will reverse engineering of file systems impact the protection of intellectual property, personal data or secu- rity of vendors and third parties? • How LEAs handle copyrighted material and security flaws in the underlying data structures? These research questions are important as currently there is little information on how law enforcement handle copy- righted material. What are, if any, their obligations to report security flaws? How are methods and tools for investigation used and where in the current legal framework law enforce- ment are facing challenges? Can investigation methods be dis- closed in trial without causing damage to different stake hold- ers unrelated to the investigation? Static-code analysis is used to scrutinize the executable code (machine code) or the source code 4 in order to under- stand the logic of the program function (what the function is meant to do) [1, p. 268] . We continue with the previous exam- ple; we are trying to identify and understand the logic of the program used by the driver to create the file. We use a special tool (disassembler) to convert object code into a more under- standable language—assembly language—and we attempt to identify the driver’s logic and functionality. 1.2. Definition and RE methods The use of reverse engineering techniques is permissible in order to determine the ideas and principles that underlie the functional elements of the software. This is usually done by examining its external inputs and outputs or testing the pro- gram/tool for errors and security flaws. Reverse engineering is defined as a “process of analyzing a subject system to iden- tify the system’s components and their interrelationships“ [16, p. 15] .The purpose of it is not replication of a program, but examination of its code, performance, or activity traces. [22] Runtime analysis is the examination of executable code (machine code) by observing the instructions as the program executes, normally using special software called debuggers, or tools that register which files have been accessed, or moni- tor network activity [4] . The debugger also converts the object code into more understandable assembly language. This definition, however, requires a more detailed descrip- tion of how reverse engineering is actually performed in re- spect to file systems. The file system is not software, but rather a set of logical structures, which are stored on virtual or phys- ical storage and are largely independent of the operating sys- tem. The file system driver handles the storage and retrieval of files and directories. The file system is similar to a database management system rather than particular database content. Therefore, the provisions for reverse engineering of programs do not apply and, as a logical schema, and not as an instance of a database, file systems are not copyright protected per se . However, they can be protected as software-related inventions and are subject to trade secret, patents and licensing [51] . On There are several issues with RE techniques when used for forensic tasks in investigation. Black-box testing is a RE method which does not infringe intellectual property rights (IPRs), but requires experiments to be scientifically validated. The other methods may reveal IPR protected information since they are studying the source and object code of the FS driver functionality (looking inside the black box). Although existing decompilers can convert object code to source code computer law & security review 46 (2022) 105725 3 Fig. 1 – Black box testing. Fig. 2 – Overview of the impact of reverse engineering for law enforcement purposes. Fig. 1 – Black box testing. Fig. 2 – Overview of the impact of reverse engineering for law enforcement purposes. 1.3. Organisation of this paper In Section 1 we have introduced the reader to reverse en- gineering of file systems, and the impact reverse engineer- ing may have on revealing trade secrets, intellectual property rights, and security flaws. A brief description of the method used is given in Section 2 . In Section 3 we provide the litera- ture review of digital forensic studies in reverse engineering and legislative frameworks. Section 4 discusses the findings related to the research questions. In Section 5 we identify ad- ditional emerging challenges, while in Section 6 we describe the recommendations related to the research questions and further work. 5 35 U.S. Code §154, a-4: “A copy of the specification and drawing shall be annexed to the patent and be a part of such patent.” 1.2. Definition and RE methods they may not be accurate [28, p. 129] and do not interpret the intended purpose of structures for evidence purposes. to be tested for its accuracy, which also requires reverse engi- neering. Most file systems used by Macintosh, Windows, etc. are proprietary and protected by copyright, patent or trade secret laws. Microsoft’s exFAT file system is a patent-protected file system [72] , while older file systems such as NTFS are copy- righted and mainly undocumented by Microsoft. When a file system is protected by patents, then the specifications need to be added to the patent application 5 [70] meaning that some metadata structures will be published by the vendor leading to a decreased need for reverse engineering. Unfortunately, even if parts of a file system are protected by patents, not all struc- tures are documented. A file system may be updated after the patent application and therefore the specifications may not be correct or complete. These undocumented structures may be relevant to the criminal investigation and require reverse en- gineering. The documentation given by the FS vendor needs 3.2.1. EU vs US copyright law 3.2.1. EU vs US copyright law In the European Union, according to Art. 5 (3) of Directive 2009/24/EC (Software Directive) [31] “a person having a right to use a copy of a computer program shall be entitled, with- out the authorisation of the right-holder, to observe, study or test the functioning of the program in order to determine the ideas and principles which underlay any element of the pro- gram if he does so while performing any of the acts of load- ing, displaying, running, transmitting or storing the program which he is entitled to do”. In addition, European laws support interoperability between computer systems and software, and some proprietary software components could be reverse engi- neered in order to enable other or open-source services. This was common practice by the development of the NTFS driver for Linux [45, p. 16] . Also, interface specifications are consid- ered not protected by copyright in Europe. Art. 6 of the Soft- ware Direcrive defines decompilation 6 as a legally-compliant practice “where reproduction of the code and translation of its form [...] are indispensable to obtain the information neces- sary to achieve the interoperability of an independently cre- ated computer program with other programs”. However, the Directive does not address disassembling 7 as a separate issue. Sanger [64] describe that current Juris Doctor (JD) curricula do not contain education in forensic sciences, which they pro- pose should be mandatory in legal education. The lawyers, judges, and prosecutors must have sufficient knowledge on scientific validity of digital forensic methodology and the reli- ability of the digital evidence. Otherwise, they will not be able to perform the legal evaluation of the evidence, or to decide which investigation methods or trade secrets could be dis- closed publicly in trial and which need to be examined under non-disclosure agreements. Twelve years ago Garfinkel [37] predicted that the lack of standards in the extraction of information from cell phones, video game consoles, telecommunications equipment, etc would become a major challenge. The techniques used to pro- tect intellectual property make these systems more difficult to acquire and analyse. He describes that many law enforcement resources are used for reverse engineering without following a standard set of procedures. Le Khac et al. Method and assumptions This paper is a multidisciplinary literature review conducted by legal scholars, law-enforcement, and digital forensics spe- 5 35 U.S. Code §154, a-4: “A copy of the specification and drawing shall be annexed to the patent and be a part of such patent.” computer law & security review 46 (2022) 105725 4 cialists, with the objective being to identify gaps between legal provisions and current digital forensic practices. The assump- tion is made that reverse engineering is a necessity in the de- scribed scenarios and many law enforcement agencies rely on the results of reverse engineering methods, which require le- gal evaluation. The interdisciplinary nature of the paper re- quires a level of abstraction of potential legal issues. Since both IPRs and criminal procedure are under national legis- lation, with limited international standardization, the paper doesn’t examine concrete jurisdictions. Instead it aims at rais- ing high-level, legal-technical problems. the scope of their authorization”considering their negative ef- fects on privacy and security. Polanski [58] analysed whether reverse engineering is permissible under EU law. He posits that any computer program is protected as a literary work, and it is the high level source code and the object code that is pro- tected. The graphical user interface and file formats can also be protected, but the functionality, ideas or principles of a pro- gram are not protected by copyright law. He also describes that observation of a legally acquired program in order to under- stand the ideas and principles utilized by the program is al- lowed, and that reverse engineering cannot be limited by a contract either. 6 The process of converting a binary program to readable high level source code. 7 Similar to decompilation, but convert to an assembly code which is lower than the normal source code. 3.2.1. EU vs US copyright law [49] describe vehicle forensics and address the challenge that obtaining proprietary information about the vehicle may be impossible due to intel- lectual property protection, and sharing information of dis- covered vulnerabilities can be used to compromise the safety of the vehicles. Further they describe tools, such as iVe [54] , which are able to extract physical or logical data using the ODB II port (On-Board Diagnostic port) [75] , but it is uncertain how this proprietary extraction is performed. They may use the in- fotainment system to gather the data, or they may analyse the QNX or VxWorks file systems. They also found that there was no forensic tool available to extract information from immobi- lizers or other electronic control units (ECUs), and they relied on using the third party diagnostic tool VCDS [62] to gather in- formation from a 2012 VW Golf. Since it is necessary to turn on the car in order to use the VCDS tool to gather all information, important GPS location information is overwritten. In USA, software protection laws introduce some broad re- quirements on reverse engineering such as the need for au- thorisation (copy of the software must be legally obtained or the copyright owner must permit the reversing) and lawful- ness (only according to the exemptions provided by law) [60] . In addition, the tools used must be legal and source code re- production and further use of implementation details are pro- hibited. The Digital Millenium Copyright act (DCMA) includes ex- ception for lawfully authorised law enforcement, intelligence, and national security purposes, for making software interop- erable, and for encryption and computer security research un- der certain conditions [60] . Currently, most cell phone reverse engineering is per- formed by DF researchers and/or commercial digital forensic tool providers. The DF practitioners in law enforcement de- pend on these digital forensic tools [36] . The detailed method- ologies implemented are hidden as trade secrets within closed source tools. Adams [3] summarizes US case law to examine the legal issues which arise from digital forensic tool devel- opment. He argues that it is desirable for “forensic tools to be designed to avoid having them used in ways that exceed 7 Similar to decompilation, but convert to an assembly code which is lower than the normal source code. 6 The process of converting a binary program to readable high level source code. 8 Daubert v. Merrell Dow Pharmaceuticals, Inc., 509 U.S. 579, 1993. The Daubert criteria was further elaborated in General Electric Co. v. Joiner 522 U.S. 136 (1997), and Kumho Tire Co. v. Carmichaet 526 U.S. 137 (1999). Althought Daubert standard is jurisdiction-specific, it had a significant impact in the digital forensics domain and is considered a de facto standard. 3.2.3. Trade secrets law 4.1.2. Peer review challenges There are no universally recognized standards for evaluat- ing the reliability of digital evidence. However most countries have requirements for scientific validation of digital foren- sic findings. For example, the UK has complex reliability re- quirements, exclusionary rules, and follow ISO accreditation [27,66] . Others, such as Germany rely on formal accreditation and court-appointed experts [12] . In Norway, due to the early involvement of the court in the case, the judge has full dis- cretion on the mandate and evaluation of the expert evidence where, for digital forensics, no formal requirements or quality controls are established [63] . The US Supreme Court formu- lated the Daubert standard 8 that requires the forensic theory or technique to be tested, peer-reviewed, generally accepted in the scientific community, and having known error rates. Some reverse engineering of file systems techniques and the techniques of using metadata structures to recover files are published and peer-reviewed digital forensic methodologies. [41,56,57] However, such RE techniques do not fully meet the Daubert standard for scientific validity in court since the RE methods are often not tested and no error rate is known. 3.2.3. Trade secrets law European Directive 2016/943/EU [32] requires all countries to implement measures to protect trade secrets and to set obli- gations for law enforcement to protect confidential informa- tion during legal proceedings (Art. 9 in conjunction with Rec. 18). By contrast, according to US Trade Secret Law, an individual shall not be held criminally or civilly liable under any Federal or State trade secret law for the disclosure of a trade secret that–(A) is made -(i) in confidence to a Federal, State, or local government official, either directly or indirectly, or to an attor- ney; and (ii) solely for the purpose of reporting or investigating a suspected violation of law; or (B) is made in a complaint or other document filed in a lawsuit or other proceeding, if such filing is made under seal.” [71] 3.2.2. Patent law A software patent does not protect source code, but rather the functionality embedded in it. Kumar and Kumar [48] de- computer law & security review 46 (2022) 105725 5 scribe that a governmental authority awards exclusive rights (patent) for a fixed number of years to the individual who first discloses the invention. The patent is an intellectual property right in exchange for full disclosure of the invention. Under the agreement on Trade-Related Aspects of Intellectual Prop- erty Rights (TRIPS), signatory countries are obliged to make patents available for any technological invention. Important issues for software patents are where the boundary of what is patentable lies, and whether software patents discourage invention. They compare UK and US legislation and conclude that it is unclear what software categories are patentable. A patent emphasizing hardware rather than software is more likely to be patentable. When it comes to India and China soft- ware will need to be implemented in hardware or presented in material form in order to be considered patentable. The UK, India and China all require a technological application, while the US requires usefulness. The authors [48] describe that software is protected as works of literature, and patentable as a technical invention. Copyright protection does not pro- tect ideas, while patents do. They are concerned that the monopoly effect of patents may block progress and new in- ventions. Moreover, technological inventions are often a com- bination of one or more patents and other non disclosed tech- nologies, which may still require reverse engineering of the system. scribe that a governmental authority awards exclusive rights (patent) for a fixed number of years to the individual who first discloses the invention. The patent is an intellectual property right in exchange for full disclosure of the invention. Under the agreement on Trade-Related Aspects of Intellectual Prop- erty Rights (TRIPS), signatory countries are obliged to make patents available for any technological invention. Important issues for software patents are where the boundary of what is patentable lies, and whether software patents discourage invention. They compare UK and US legislation and conclude that it is unclear what software categories are patentable. A patent emphasizing hardware rather than software is more likely to be patentable. When it comes to India and China soft- ware will need to be implemented in hardware or presented in material form in order to be considered patentable. 3.2.2. Patent law The UK, India and China all require a technological application, while the US requires usefulness. The authors [48] describe that software is protected as works of literature, and patentable as a technical invention. Copyright protection does not pro- tect ideas, while patents do. They are concerned that the monopoly effect of patents may block progress and new in- ventions. Moreover, technological inventions are often a com- bination of one or more patents and other non disclosed tech- nologies, which may still require reverse engineering of the system. 4.1.1. Interpretation challenges Horsman argues that “tools for parsing of a file system may focus on displaying file and folder content to a user and inac- curately interpret file system metadata possibly due to insuf- ficient testing” [44] . Reverse engineering of file systems is used to understand metadata structures, and can be used to: • Discover malicious activity • Understand and interpret the data structure • Recover user data Alternate Data Streams (ADS) in the NTFS file system are known to be exploited to hide malicious content. The tech- nique of using tools that identify ADS have already been peer reviewed and published in the Digital Forensic commu- nity [10] . Even though not explicitly mentioned, since ADS was undocumented, the authors of tools supporting ADS had to initially perform reverse engineering or build on previous re- verse engineering efforts. Since file systems consist of metadata structures describ- ing the file system and the files and directories it contains, previous research has tried to interpret these undocumented structures within closed source file systems by using reverse engineering [40,41] . 4.1. Technical challenges with reverse engineering in investigations In a similar vein, if LEA develop methodology to interpret file systems (in relation to a criminal case), that are unknown or unsupported by existing DF tools, it is a form of applied re- search and the methods used must be peer-reviewed in or- der to be validated and reused by others. Publishing of tech- niques in scientific journals is the preferred method to do Although the legal provisions for interoperability and testabil- ity of software generally allow for reverse engineering, laws in the US and Europe are broadly written and do not take existing methods into account, or sufficiently address new application of reverse engineering for digital forensics and evidence ac- quisition purposes. Further we argue that LEAs depend on sys- tem vendors and DF tool vendors. In order to overcome those dependencies LEAs must rely on reverse engineering. We ar- gue, that current legislation does not reflect the full complex- ity of this new context, or the legality of different methods and tools used. computer law & security review 46 (2022) 105725 6 with respect to intellectual property, trade secrets, system se- curity, and data protection. so. However, public disclosure may create serious disadvan- tages. Firstly, criminals may misuse this information to break into systems, or to avoid prosecution by utilising anti-forensic techniques based on information found in these publications. LEA might need to protect their methods by keeping them confidential and reuse them in further investigations. If the LEA method describes weaknesses in the file system, vendors might update in order to prevent RE by others. Internal valida- tion might lack impartiality or competence, while no valida- tion can introduce inaccuracies in all investigations where the technique is subsequently used. Currently, there are no com- munication channels or independent bodies where applied re- search performed by LEA can be peer-reviewed for scientific validity and disseminated among LEAs without public disclo- sure. 4.2.1. Intellectual property challenges In the EU the source and object code of an original computer program are protected as literary work. [ 31 , Art. 1] Software copyright is mostly restricted to direct reproduction of the source or object code. Art. 1 (2) of the Software directive must be interpreted that it does not provide protection in cases of non-literal infringements [55, ch 9 para 9.4] . Such infringe- ments occur when the logic, structures or data formats of a software are reproduced. 4.1.3. Method and tools challenges 4.1.3. Method and tools challenges Understanding the inner workings of a closed source file system is a time consuming process for digital forensic re- search [67] , and it is difficult to prove that the interpretation of the file system structures are complete and accurate. It should be stressed that the metadata structures identified from file system reverse engineering may not be exactly what exist on disk [14, p. 199] . In order to acquire digital evidence, LEAs must be able to test if the DF tool correctly interprets the file system, and in the case of errors and uncertainties to validate the re- sults with reverse engineering techniques. Numerous skills are required to perform reverse engineer- ing of file systems including interpretation of hexadecimal and C structures, debugging, assembly, programming, knowl- edge of existing file system structure, performing reliable ex- periments, hypothesis testing, etc. These limitations in the forensic tools and the lack of RE knowledge by the examiner could result in loss of evidence for the investigation. For instance, Microsoft‘s Resilient file system released with Windows 8 and Windows Server 2012 has a completely different storage engine than previous Mi- crosoft file systems. It can manage vast amounts of data and has improved security and data integrity capabilities which reportedly limits the current forensic tools’ ability to interpret it [56] . Apple implemented the new APFS file system in 2017, and at the time of release digital forensic tools had no support for APFS. The missing APFS support had a serious impact on criminal cases that included devices using the APFS file sys- tem, meaning many iOS and MacOS devices could not be in- vestigated [41] . Current digital forensic tools do not support vehicle file systems such as QNX or VxWorks [49] . In the ab- sence of protocols to track vendors‘ changes in system design, forensic examiners rely on reverse engineering or manual in- spection of low-level processes in order to investigate the data, detect running malware, interpret underlying data structures for data recovery, or to create / improve forensic tools. Another possibility for law enforcement to lawfully per- form reverse engineering is under the interoperability exemp- tion. Art. 6 in conjunction with Rec. 15 of the Software Direc- tive [31] permits the use of reverse engineering e.g. 4.1.3. Method and tools challenges studying file systems in order to create interoperable DF tools or to vali- date the accuracy of existing ones. CJEU considered the refusal of Microsoft to provide sufficient interoperability information regarding its operating system a violation of competition and antitrust laws. The court held that Microsoft had a special re- sponsibility to supply and authorize the use of interoperabil- ity information to others in order to achieve a high degree of interoperability that the circumstances required and not to hinder effective and undistorted competition in the Common Market [18] . Since then, Microsoft has published a substantial amount of information pertaining to protocols, APIs, etc. to enable the development of compatible software by third par- ties [53] . Moreover, the Software Directive limits contractual restrictions on reverse engineering to the extent decompiling is concerned [65] . The legal protection of interoperability is of relevance for developing and testing digital forensic tools by law enforcement because more information about closed source file system will become available to support develop- ment of alternative open source file system drivers. However, there is a difference between reverse engineering for interop- erability and reverse engineering for investigative purposes since the former is focusing on creating a full implementa- 4.1. Technical challenges with reverse engineering in investigations In this respect, the Court of Justice of the European union (CJEU) gave a landmark decision in the SAS case [17] . The court declared that logic, algorithms, programming languages and data formats are not protected by copyright. In a further judgement, the court also stated that under Art. 5 (1) of the Software Directive, the lawful pur- chaser of a program can decompile it in order to correct errors in its operations without the authorisation of the copyright holder [19] . Consequently, black-box testing does not impose a threat to intellectual property rights and trade secrets, be- cause the forensic testing is performed without directly look- ing at source or object code. It is lawful and cannot be limited by a contractor licence agreement [58] . However, black-box RE might introduce inaccuracies and questions about the reliabil- ity of this method. For static-code and runtime analysis inves- tigators must work with debuggers, decompilers, or disassem- blers and use information from previous reverse engineering of the file system or similar open source work. Some of those RE methods may reveal IPR protected information since they study the source and object code of the FS driver functionality. Others may fall under the exemption of Art. 5 (3) the Software Directive as they aim to observe and study logical metadata structures and does not reproduce the source code. 4.2.3. Challenges with trade secrets disclosure However, a recent judgement in Oracle v. Google extended copyright protection to Java declaring code, which is considered an aberration of the global consensus for interop- erability [34] . Even when reverse engineering for law enforcement is per- missible under EU and US law, some questions remain unad- dressed by the legislator e.g. law enforcement obligations in relation to disclosure of RE results to the defence, discovered sensitive or confidential data, or found security vulnerabili- ties in systems and their cross-jurisdictional effects. The le- gality of RE tools and methods or their impact on security of systems is largely unaddressed. In addition, some LEAs buy digital forensics tools from vendors who’s responsibilities for reverse engineering are not clarified. As exemplified further, in most cases jurisdiction specific rules apply. 4.2.2. Challenges with IPRs disclosure In most jurisdictions, a judge can issue a warrant for disclo- sure of copyright protected information for the purpose of criminal investigation. In the UK for example, the PACE pro- duction orders [42] allow the court to oblige an organisation to provide confidential content data (e.g. confidential files or e-mails) to law enforcement. The provision refers to infor- mation, which the organization has a duty to keep confiden- tial, but it remains unclear if this includes security and copy- righted data, which the organisation has a right to keep con- fidential. Consequently, the right holder can limit the disclo- sure to the warrant‘s scope or challenge it in court. However, more commonly LEA can lawfully acquire the device (com- puter or mobile) proceed to a warrant and later on, during forensic analysis, establish the need of reverse engineering. The RE will be considered as digital evidence acquisition or analysis where no additional warrant will be necessary. In this case the right holder will not be able to protect their system, and it will be up to the LEA to follow general requirements for RE and to protect confidential information. Detailed docu- mentation of the forensic methodology is not only important for its reliability validation but moreover for proving that the digital forensic examination was limited to the search warrant scope and did not violate privacy and data protection [3] . How- ever, digital forensics methods are often poorly documented [43] , while search warrants are broadly formulated and do not limit the scope of the digital investigation [69] . 4.2.3. Challenges with trade secrets disclosure 4.2.3. Challenges with trade secrets disclosure 4.2.3. Challenges with trade secrets disclosure Reverse engineering for investigation purposes might have further impact on trade secret protection. To exemplify the issues we discuss relevant legislation in Germany and the United States. Germany is an example of a European coun- try that implemented the Trade secret Directive and regulates law enforcement activities in such context. According to cur- rent legislation, reverse engineering by LEA is considered un- lawful, but a major legislative change was introduced recently. The published Draft Bill [38] , in accordance with the Trade Se- cret Directive, states that reverse engineering is a permitted means for obtaining trade secrets when one of the follow- ing conditions is met: the product or object (i) has been made available to the public, or (ii) is lawfully in the possession of a party who is free from any legally valid duty of confidentiality. Thus, as long as the product or object has not been made pub- licly available, it remains possible to contractually prohibit re- verse engineering to the extent permitted by law. This is con- trary to the Software Directive as well as the German Copy- right Act, which limit contractual restrictions on reverse engi- neering to the extent decompiling is concerned [65] . Reverse engineering (i.e. analysis and dismantling of products for the decoding of secrets) is now generally permissible according to the new regulation in Section 3 para. 1 no. 2 GeschGehG 9 - but still only within the restrictions of the laws against the unfair competition and the intellectual property laws [46] . A possible inadequacy of the system has emerged since, in crim- inal proceedings, the defence counsel has to be given access to files, but there is no explicit provision requiring them not to disclose the trade secrets contained therein [7] . The issues however, have been assessed by the German Supreme Court, which stated that the defence counsel can disclose to its client only the information that is indispensable for a proper de- fence [7] . In US perspective similar exceptions are found in copy- right law. 17 U.S. Code §1201 (e) provides immunity for law enforcement authority who are lawfully authorized to circum- vent technology means for copyright protection, including for the purpose of identifying and addressing security vulnerabil- ities and 17 U.S. Code §1201 (f) codifies an interoperability ex- emption [60] . 9 German Act on the Protection of Trade Secrets (Gesetz zum Schutz von Geschäftsgeheimnissen vom 18. April 2019 (BGBl. I S. 466)). 4.2. Legal challenges with reverse engineering for law enforcement purposes Computer systems are a mixture of copyrights, patents, li- censes, and freely accessible code. Reverse engineering of data structures is not regulated specifically for the purpose of dig- ital evidence acquisition and analysis, but has implications computer law & security review 46 (2022) 105725 7 tion of the file system on an unsupported platform, while the latter is focusing on understanding the meaning of metadata structures mainly related to files. Arguably, information pro- vided by vendors for interoperability is insufficient and does not meet the LEA’s need for correct data interpretation. In the absence of legal provisions to compel software vendors to pro- vide such information for law enforcement purposes, reverse engineering remains the only possible solution, although very burdensome and not always fully accurate. transparent are poorly addressed in the literature and in leg- islation. The aim of greater transparency in digital forensics must be combined with mitigation of the negative impact dis- closure can have on trade secrets and system vendors’ copy- rights. 4.2.3. Challenges with trade secrets disclosure Problems with what is permissible after the device or electronic material is seized, and to what extent digital forensic techniques must be Under US Trade Secret Law law enforcement are exempt from the protection of trade secrets. The Parties have the right to disclose in confidence trade secrets to Federal, State, and lo- cal government officials, or to an attorney, for the sole purpose of reporting or investigating a suspected violation of law. Due to the peculiarities of competition law proceedings in the US when compared to the European model, the discovery process permits a party to demand disclosure of relevant information and potential evidence from the other party. This includes in- formation on copyrighted material or trade secrets that are relevant for litigation. However, a party from whom discovery is sought may move the court to issue a protective order re- quiring no public disclosure or to reveal only in a limited man- ner a trade secret [7] . Nothing in this Agreement is intended to conflict with 18 U.S.C.§1833(b) or create liability for disclo- sures of trade secrets that are expressly allowed by 18 U.S.C. computer law & security review 46 (2022) 105725 8 and the behaviour of the tools used to perform these oper- ations [52] . Current practices and legislation are not able to overcome system vendor and DF-tool vendor dependencies, which undermines the scientific validity of the performed dig- ital forensics. Secondly, DF-tools are multi-purposed and are used in many different security, intelligence and investiga- tion contexts. Commercial tool vendors do not necessarily ac- count for the higher standards LEA need to meet in respect to reliability of methods for court proceedings. Marshall and Paige [52] conducted a questionnaire asking DF-tool vendors if they consider law enforcement (customer) requirements for their tools, and 12 of 14 vendors were confused about the meaning of law enforcement requirements. Moreover, new file systems or versions remain unsupported by DF tools for sig- nificant time periods [44] , while investigations can not be dis- missed until a tool to interpret the data is created. There- fore, the digital forensic examiner cannot rely solely on digital forensic tools. Carrier argues that “software developers must... release their source code if it is used to generate evidence. If a developer is unwilling to do so [...this] can be a factor when purchasing an analysis tool” [13] . 4.3. Vulnerabilities disclosure During investigations LEA might discover security vulnerabili- ties. Currently, there is little information what are, if any, LEA’s obligations to report security flaws. In the absence of a swift and secure communication process they might have little in- centive to inform vendors, since this may impact law enforce- ment’s ability to acquire data. The more difficult it is to ac- quire the data, the more resources law enforcement need to use on acquisition, which will make less time available for analysis. Moreover, device producers gradually increase secu- rity and privacy measures in their devices [6] and, in a worst case scenario, the police might not be able to gain access to pertinent data, which then threatens public safety. 4.2.3. Challenges with trade secrets disclosure As long as forensic tool ven- dors are implementing secret techniques in their tools, there will always be a need to perform reverse engineering of file system drivers and file system structures in order to identify new or updated structures that may have an impact on the investigation. In addition, the investigator could detect errors or forensic tools performing poorly on new data structures, which might require reverse engineering not only for the evi- dence file system, but also reverse engineering parts of the DF tool. §1833(b). Wexler [74] argues against trade secret privileges in criminal cases, because if information on how evidence has been gathered is excluded from the case, this could jeopardize the accused‘s right to a fair trial. The ambiguous regulation in the US, as Wexler argues, leaves an open question whether trade secret privileges apply to criminal cases. 4.2.4. FS vendors’ copyrights 4.2.4. FS vendors copyrights When investigating a criminal case the aim is to find enough relevant information, which may have evidentiary value, to define hypotheses and then to try to falsify them in order to find the most likely hypothesis [33,59] . If such information is unavailable because of intellectual property rights protec- tion, then this may impact the accuracy of the hypothesis testing, and decrease the reliability of identifying the most likely hypothesis. This may impact the right to a fair trial, because relevant evidence can not be accessed. This is also why law enforcement is facing challenges with intellectual property protection of file system vendors and digital forensic tool vendors. Some of the file systems and underlying func- tionalities are proprietary. The file system driver, the man- ager, is the software responsible for all the file system activ- ity which it performs when users create, update, delete, move, or copy files. This driver can be a mixture of propriety and open-source code, and could be changed completely due to in- creased storage and processing capabilities of new computer systems when the vendor updates the driver. Law enforce- ment does not have access to the low-level changes of each file system version, since vendors’ keep this information secret in order to preserve the integrity and security of their products, but more importantly protect customers’ data and other con- fidential information. The vendors’ objective is always to im- prove performance and customer experience [40, p.7] , which can cause inaccuracy in digital forensic tools parsing the file system. Therefore, vendors may be reluctant to cooperate with LEA. Reverse engineering by LEA might lawfully reproduce IPR- protected information and trade secrets but the further use of such information depends on the jurisdiction specific rules. 5. Emerging challenges Due to the rapid growth of data and complexity, digital foren- sics requires more computational assistance and automation [15] . The introduction of automated, complex tools in inves- tigations will sharpen the identified challenges with security, privacy, intellectual property, or methodology validation and disclosure. The lack of a regulatory framework to ensure ac- countability and reliability in big data analytics for investi- gations is already outlined as an emerging challenge for law enforcement [9] . Further, in mobile forensics most investiga- tors rely on the digital forensic tools provided by commer- cial vendors. The current practice of “push-button forensics” and complex closed-source forensic tools [44] creates tool de- pendencies by LEA where they only report tool results, but can not sufficiently test the correctness of the tool interpre- tation in each case. DF tool vendors may implement auto- mated analytic features to assist the investigator. How such functionalities are implemented is kept secret. In order to un- derstand and validate the methods utilised by these digital forensic tools, the only option currently is to perform reverse engineering. Therefore, it is good practice to comply with the ABC model: “Assume nothing, Believe nothing, Challenge and Check everything“ [33] . However, this is not necessarily done by practitioners because they do not have the time or knowl- edge. It’s also legally unclear if they are allowed to perform reverse engineering and which methods they are allowed to use. Therefore, most investigators rely on the reverse engi- neering performed by commercial tool vendors, and use the tool that solves the investigative issues. It is also question- able if the commercial vendors are allowed to perform this reverse engineering. In the absence of robust formal proce- dure for validation, investigators can not avoid tool depen- dencies, which undermine the scientific soundness of the examination. File system encryption is a real challenge for law enforce- ment, especially since some devices have hardware encryp- tion (for instance the Mac with the T2 chip has hardware en- cryption support [5] ). Without the encryption key, as far as we know, it can not be decrypted within a reasonable time frame. This is of course good for privacy. However, law enforce- ment need access to data when investigating serious crime. It has also been suggested that vendors of devices should add back doors to their systems. 4.2.5. DF-tool vendors’ copyrights Without sufficient stan- dards and documentation of how LEA examined the device after decryption, it will be difficult to prevent abuse of those broad powers and excessive intrusion in individuals’ private sphere, including those not related to the investigation. In addition, file system encryption are measures taken by the vendors to protect personal data. The European data protec- tion supervisor emphasises that the “decryption, reverse en- gineering or monitoring of communications protected by en- cryption should be prohibited [as well as...] facilitating ’back- doors’ [30, p.36] . Interpol recently pointed out that tension be- tween privacy and digital forensics might lead to “the limita- tion of utility of digital evidence by means of regulation and legislation” [61] . recommends adoption of procedures by all LEA, where they have: (i) an obligation to report vulnerabilities; (ii) may only temporarily restrict knowledge of a vulnerability; and (iii) an oversight body ensures compliance. Currently, in the absence of a legal framework which reg- ulates the relation of digital forensics and evidence with data protection and security laws, the process imposes a higher burden on the forensic examiner, to make decisions on dis- closure of methodology, sensitive data or security vulnerabil- ities. Arguably, legal arguments with 3 rd rd parties for security protection or investigation methods disclosure will reduce the efficiency and reliability of digital forensics. 4.2.5. DF-tool vendors’ copyrights DF-tool vendors also rely on reverse engineering of FS and pro- tect their methods in order to have competitive products. Most of the digital forensic tools are closed source, and how the vendors have implemented the methods used is unknown to law enforcement. However, the use of closed-source forensic tools becomes increasingly problematic for law enforcement. Firstly, law enforcement can not blindly trust the accuracy of the tools, and best practice requires validation [44] , which might affect copyrighted functions of the DF tool. Adams [3] is summarizing legal practice in the US and the legal is- sues related to the development of digital forensic tools. He states that reliability of DF tools can not be assumed due to many bugs, errors, and timestamp inaccuracies, while defence lawyers might request to review the source code of the foren- sic tool. Marshall and Paige further explain that it “should also be borne in mind that updates to hardware and software may have no apparent effect on system behaviour as far as a typ- ical user is concerned, but may dramatically change the way in which internal processing is carried out and data is stored.” This impacts both the ability to recover and interpret data Sharing methodologies or knowledge of newly discovered data structures with the court or the wider security and digital forensic communities might expose system‘s sensitive infor- mation to competitors and criminals. Therefore, security ex- perts already discuss the danger of forensic data leakage and harden software against security flaws in file or operating sys- tems. For example, the HFS journal writes a cache of changes to the file system allowing encryption key and file content re- covery [76, p. 273] , which could be of great value to forensic ex- aminers, but is also considered providing insufficient security of the file systems unallocated space by the software devel- opers. A report on government disclosure processes in Europe stated that only a limited number of countries have a trans- parent procedure for vulnerability disclosure [39, p.86] . Simi- lar to the US Vulnerabilities Equities Process (VEP) the report computer law & security review 46 (2022) 105725 9 need for a legal framework to mitigate the risks of government hacking [26] . They recommend the adoption of guidelines for handling digital evidence, hacking only for “serious crimes”, transparent reporting, and binding requirements for vendors of government hacking tools [24] . 4.4. Circumvention of encryption/security and personal data leakage Often reverse engineering of file systems for digital forensics purposes includes circumvention of security measures or en- cryption [8] , in order to make the data available. One of the biggest concerns about reverse engineering of file systems for evidence acquisition is that it can compromise system secu- rity, exposing vendor secrets and customer data, which is oth- erwise encrypted at the application layer. The dispute around governmental access to encrypted and protected data drew public attention with Apple‘s refusal to compromise its iPhone security in response to an FBI request to get access to the phone belonging to one of the San Bernardino shooters [50] . The iPhone‘s data was encrypted, but the software control- ling the phone was not, which was a security flaw and the FBI requested this software to be substituted by Apple with one which allows rapid password guessing. It is interesting that the FBI withdrew the request because, according to anony- mous sources, the Israeli company Cellebrite assisted the FBI in opening the phone [68] . Cellebrite is specialised in develop- ing digital forensics tools and solutions. The US Court did not allow disclosure of the forensic examination and data recov- ery as “national security secrets and ‘intelligence sources or methods’ that are exempt from disclosure under the Freedom of Information Act” [21] . 5. Emerging challenges If the trial safeguards for admissibility and reliability of evi- dence are sufficient to scrutinize digital forensic techniques and tool results remains questionable [27] . One option may be the examination of the RE method under a non-disclosure agreement in closed door court proceedings for cases where vulnerabilities or trade secrets are disclosed. Another option is non-disclose of details of trade secrets or vulnerabilities. Both solutions will limit the right to information and ac- countability obligations for law enforcement. On the other hand the secrecy may protect third party copyrights, and mit- igate the danger of criminals adapting their modus operandi ac- cording to disclosed law enforcement methods and disclosed vulnerabilities. • Legislators must reconsider the impact of digital forensic methods on intellectual property, data protection and se- curity of commercial systems and to regulate the permissi- ble techniques and tools which do not jeopardize existing or future security and allows cross-examination of the evi- dence for trial [3,61] . A regulation at European level can har- monize and set minimum standards for methods, valida- tion, and disclosure which will support LEA’s cooperation in all countries [39] . Further research is needed to establish the legal basis and scope of such legislation. • We argue that file system vendors must sub-license and disclose the details regarding structures important for in- vestigation. This requires information on how data is logi- cally stored and how data structures should be interpreted for law enforcement purposes. Such measures will signif- icantly reduce the resources needed for reverse engineer- ing of file systems, which will remain reserved only for tool testing, validation and investigation. The approach could be further generalized to other storage solutions such as undocumented instances of databases used by applica- tions. • We argue that file system vendors must sub-license and disclose the details regarding structures important for in- vestigation. This requires information on how data is logi- cally stored and how data structures should be interpreted for law enforcement purposes. Such measures will signif- icantly reduce the resources needed for reverse engineer- ing of file systems, which will remain reserved only for tool testing, validation and investigation. The approach could be further generalized to other storage solutions such as undocumented instances of databases used by applica- tions. 10 See the requirements by European Court of Human rights in Allan v. the United Kingdom, judgement of 5 November 2002, para 47; Bykov v. Russia [GC], 4378/02, 10 March 2009; Dragojevi ´c v. Croa- tia, judgement of 15 April 2015, para 129. 5. Emerging challenges • Digital forensic tool developers must have an obligation to provide LEA with information necessary for scientific vali- dation of the tools and their interpretation of the file sys- tems. • New forensic methodologies including reverse engineer- ing or other types of applied research during investigations must be documented according to formalized procedures in order for their scientific validity to be evaluated. This re- quires a level of transparency in digital forensics which is not reached currently due to closed-source tools and meth- ods. • Law Enforcement units need to test and peer-review their own techniques and methods [29] . It is not efficient that ev- ery lab perform their own testing of the same tools or tool versions. However, a detailed public dissemination of the techniques and methods may be exploited by criminals. It could be sufficient in more complex cases for another forensic examiner to reproduce the results and validate the tools. • Law Enforcement units need to test and peer-review their own techniques and methods [29] . It is not efficient that ev- ery lab perform their own testing of the same tools or tool versions. However, a detailed public dissemination of the techniques and methods may be exploited by criminals. It could be sufficient in more complex cases for another forensic examiner to reproduce the results and validate the tools. 5. Emerging challenges This is not a good solution be- cause back doors can be exploited by criminals and third par- ties [47] and compromise privacy [2] . Even governments need to keep secret information confidential, and strong encryption is one method [47] through which this is achieved. Some ju- risdictions allow the police to compel the encryption key from suspects or their close circle [47] . Of course the disclosure of cryptographic keys also gives law enforcement access to more data than what is related to the crime under investigation, and may impact the right against self-incrimination and the right to silence. Additionally some EU countries have given LEAs hacking powers, allowing the police to hack into systems in order to gather information [47] . Authors emphasise the Further complications, are related to the trial proceed- ings. According to Art. 6 of the European Convention on Hu- man Rights, the right to a fair trial requires public cross- examination of the digital evidence, opportunity for the de- fence to challenge digital forensic findings, to assess the computer law & security review 46 (2022) 105725 10 reliability and accuracy of the evidence and to oppose its use 10 . This means that if reverse engineering is part of the forensic examination, the methodology must be documented sufficiently in the investigation in order to enable cross- examination during trial proceedings. Moreover, law enforce- ment must use legal methods and tools to reverse engineer the file system. Court and/or defense review of scientific evi- dence is considered important to ensure an equality of arms, a fair trial and a safe verdict. This is far more important, where the forensic tool and method are novel or insufficiently vali- dated [25] . The lawyers and the court need to verify the dig- ital forensic examination and to keep sub-standard scientific techniques or methods out of the court [64] . The investigator could be reluctant to testify about investigation methods or security flaws in systems, and unfortunately we have seen in the USA the use of alternative explanations for how evidence was found, a practice known as “parallel construction“ [20,73] . 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Current legis- lation insufficiently addresses issues with sensitive method- ology disclosure, security vulnerability disclosure, applied re- search by law enforcement, or tension between IPR protection and validation of forensic methodologies. Moreover, current practices and legislation are unable to overcome system- and DF-tool vendor dependencies, which undermine the scientific validity of the digital forensic process. File systems or appli- cations are not built to comply with law enforcement needs and vendors of digital forensic tools do not necessarily con- sider the law enforcement obligations [52] . Below we list a few topics which are important to address: • Law Enforcement need training in how to scientifically test tools and perform peer-review [29] . 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https://openalex.org/W2124330529	https://eprints.qut.edu.au/220121/1/64192.pdf	English	null	Physical activity patterns among South-Asian adults: a systematic review	The international journal of behavioural nutrition and physical activity	2,013	cc-by	9,036	This may be the author’s version of a work that was submitted/accepted for publication in the following source: Ranasinghe, Chathuranga, Ranasinghe, Priyanga, Mallika Arachchige, Ranil, & Misra, Anoop (2013) Physical activity patterns among South-Asian adults: a systematic review. International Journal of Behavioral Nutrition and Physical Activity, 10, pp. 1-11. This ﬁle was downloaded from: https://eprints.qut.edu.au/220121/ This may be the author’s version of a work that was submitted/accepted for publication in the following source: Ranasinghe, Chathuranga, Ranasinghe, Priyanga, Mallika Arachchige, Ranil, & Misra, Anoop (2013) Physical activity patterns among South-Asian adults: a systematic review. International Journal of Behavioral Nutrition and Physical Activity, 10, pp. 1-11. This ﬁle was downloaded from: https://eprints.qut.edu.au/220121/ This may be the author’s version of a work that was submitted/acc for publication in the following source: Ranasinghe, Chathuranga, Ranasinghe, Priyanga, Mallika Arach Ranil, & Misra, Anoop (2013) Physical activity patterns among South-Asian adults: a systematic r International Journal of Behavioral Nutrition and Physical Activity, 1 1-11. This ﬁle was downloaded from: https://eprints.qut.edu.au/220121/ c⃝Consult author(s) regarding copyright matters This work is covered by copyright. Unless the document is being made available Creative Commons Licence, you must assume that re-use is limited to personal u that permission from the copyright owner must be obtained for all other uses. If th ment is available under a Creative Commons License (or other speciﬁed license) th to the Licence for details of permitted re-use. 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Unless the document is being made available under a Creative Commons Licence, you must assume that re-use is limited to personal use and that permission from the copyright owner must be obtained for all other uses. If the docu- ment is available under a Creative Commons License (or other speciﬁed license) then refer to the Licence for details of permitted re-use. It is a condition of access that users recog- nise and abide by the legal requirements associated with these rights. If you believe that this work infringes copyright please provide details by email to qut.copyright@qut.edu.au License: Creative Commons: Attribution 2.5 Notice: Please note that this document may not be the Version of Record (i.e. published version) of the work. Author manuscript versions (as Sub- mitted for peer review or as Accepted for publication after peer review) can be identiﬁed by an absence of publisher branding and/or typeset appear- ance. If there is any doubt, please refer to the published source. https://doi.org/10.1186/1479-5868-10-116 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 6 Keywords: Physical activity, Inactivity, South Asia, Adults Studies in the United Kingdom have shown that the risk of diabetes is 3 to 5 times higher for immigrants from Bangladesh, India and Pakistan compared with the native white Caucasian population, with an associated increased risk of complications, morbidity and mortality [3]. South Asia has the highest number of patients with diabetes and the prevalence of diabetes among adults is over 10% in many parts of the region [4]. This increased metabolic risk among South Asians appears to be multi-factorial, where unhealthy dietary habits and physical inactivity are coupled with genetic predisposition [5]. Physical inactivity increases the risk of developing abdominal adiposity, diabetes and cardiovascular disease [6]. Furthermore, physical inactivity is considered the fourth leading risk factor for global Physical activity patterns among South-Asian adults: a systematic review Chathuranga D Ranasinghe1, Priyanga Ranasinghe2, Ranil Jayawardena3 and Anoop Misra4 Chathuranga D Ranasinghe1, Priyanga Ranasinghe2, Ranil Jayawardena3 and Anoop Misra4 © 2013 Ranasinghe et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. * Correspondence: chath_r@yahoo.com.au 1Allied Health Sciences Unit, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka Full list of author information is available at the end of the article Abstract Physical activity (PA) has many beneficial physical and mental health effects. Physical inactivity is considered the fourth leading risk factor for global mortality. At present there are no systematic reviews on PA patterns among South Asian adults residing in the region. The present study aims to systematically evaluate studies on PA patterns in South Asian countries. A five-staged comprehensive search of the literature was conducted in Medline, Web of Science and SciVerse Scopus using keywords ‘Exercise’, ‘Walking’, ‘Physical activity’, ‘Inactivity’, ‘Physical Activity Questionnaire’, ‘International Physical Activity Questionnaire’, ‘IPAQ’, ‘Global Physical Activity Questionnaire’ and ‘GPAQ’, combined with individual country names. The search was restricted to English language articles conducted in humans and published before 31st December 2012. To obtain additional data a manual search of the reference lists of articles was performed. Data were also retrieved from the search of relevant web sites and online resources. The total number of hits obtained from the initial search was 1,771. The total number of research articles included in the present review is eleven (India-8, Sri Lanka-2, Pakistan-1). In addition, eleven country reports (Nepal-3, Bangladesh-2, India-2, Sri Lanka-2, Bhutan-1, Maldives-1) of World Health Organization STEPS survey from the South-Asian countries were retrieved online. In the research articles the overall prevalence of inactivity was as follows; India (18.5%-88.4%), Pakistan (60.1%) and Sri Lanka (11.0%-31.8%). STEPS survey reports were available from all countries except Pakistan. Overall in majority of STEPS surveys females were more inactive compared to males. Furthermore, leisure related inactivity was >75% in studies reporting inactivity in this domain and people were more active in transport domain when compared with the other domains. In conclusion, our results show that there is a wide variation in the prevalence of physical inactivity among South-Asian adults within and between countries. Furthermore, physical inactivity in South Asian adults was associated with several socio-demographic characteristics. Majority of South Asian adults were inactive during their leisure time. These Factors need to be considered when planning future interventions and research aimed at improving PA in the region. Keywords: Physical activity, Inactivity, South Asia, Adults Search strategy A fi d A five staged comprehensive search of the literature was conducted in the following databases; PubMed® (U.S. National Library of Medicine, USA), Web of Science® [v.5.4] (Thomson Reuters, USA) and SciVerse Scopus® (Elsevier Properties S.A, USA) for studies published before 31st December 2012. During the first stage the above databases were searched using the following search criteria. The Medline database was searched using the MeSH (Medical Subject Headings) terms ‘Exercise’ and ‘Walking’, with keywords ‘Physical activity’, ‘Inactivity’, ‘Physical Activity Questionnaire’, ‘International Physical Activity Questionnaire’, ‘IPAQ’, ‘Global Physical Activity Questionnaire’ and ‘GPAQ’. The search terms were com- bined with the names of the individual South-Asian countries; ‘Sri Lanka’, ‘Lanka’, ‘Ceylon’, ‘India’, ‘Bangladesh’, ‘Pakistan’, ‘Nepal’, ‘Bhutan’, ‘Maldives’. The Web of Science database was searched using all of the above search terms in article topic. In the SciVerse Scopus database the above terms were searched in the article title, abstract or keywords. In the United Kingdom the levels of physical activity have long been known to be lower in the South Asian people than in the general white Caucasian population [14]. The desire to walk, cycle and participate in sports and recreational activity is low among South Asians in comparison with the general population in United Kingdom [14]. Data from the Health Survey for England found that South Asians were 60% less likely than native Caucasians to meet government recommendations for physical activity [15]. Likewise, a systematic review by Fischbacher et al. (2004) examined physical activity levels among South Asians and compared it to the general United Kingdom population and noted that rates were 50–75% less in South Asians [16]. Lack of understanding about benefits, communication gap with health care professionals, cultural beliefs and lack of culturally sensi- tive facilities are some of the potential barriers for physical activity in South Asians [17]. Swaminathan and Vaz, systematically reviewed the physical activity level of children in India [18]. Although there are several stud- ies from individual South Asian countries on the level of physical activity among adults in each country, at present there are no comprehensive systematic reviews on physical activity patterns among South Asian adults residing in the region. The aim of this exercise is to systematically evaluate published work on physical activ- ity from individual South Asian countries and summarize as for a common region enabling comparison with other regions. Introduction South Asia, commonly known as the Indian sub-continent, is home to almost one-fifth of the world’s population and is comprised of many diverse ethnic, linguistic and religious groups [1]. Altogether there are 7 countries in the region namely; Bangladesh, Bhutan, India, Maldives, Nepal, Pakistan and Sri Lanka. Although there are signifi- cant cultural differences between regional countries, South Asians are an inherently high-risk group for developing abdominal adiposity, diabetes, cardiovascular diseases [2]. * Correspondence: chath_r@yahoo.com.au 1Allied Health Sciences Unit, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka Full list of author information is available at the end of the article Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Page 2 of 11 Page 2 of 11 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Search strategy A fi d In the second stage the total hits obtained from searching these three databases were pooled together and duplicates were removed. This was followed by screening of the retrieved articles by reading the article title in the third stage and abstracts in stage four. In the fifth stage individual manuscripts were screened, and those not satisfying inclusion criteria (given below) were excluded. To obtain additional data a manual search of the reference lists of articles selected in stage five was performed. Wherever possible forward citations of the studies retrieved during the literature search was traced and screened for possible inclusion. Furthermore, data were also retrieved from the search of relevant web sites and online resources. This search process was conducted independently by two reviewers (PR and RJ) and the final group of articles to be included in the review was deter- mined after an iterative consensus process. The initial literature search using the above search criteria identified the following number of articles in the respective databases; Medline® (n = 121), Web of Science® (n = 424) and SciVerse Scopus® (n = 1226). The search strategy is summarized in Figure 1. Methods mortality causing an estimated 3.2 million annual deaths (6% of global deaths) [7]. A systematic review of published studies reporting physical activity among South Asian adults residing in South Asia was undertaken in accordance with the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) statement (Additional file 1) [19]. Physical activity is defined as any bodily movement produced by skeletal muscles that substantially elevates energy expenditure [8]. It is an established fact that physical activity has many beneficial health effects. Regular (≥3 times per week) moderate intensity physical activity such as brisk walking, dancing and gardening decreases the risk of non-communicable disease [9]. Adequate phys- ical activity is also known to increase mental well being [10]. The Harvard alumni study has shown that the alumni mortality rates were significantly lower among those who were physically active, even after adjusting for other lifestyle risk factors [10]. Regular physical activity has been reported to lower blood pressure in adults with hypertension [11]. Physical activity assists in weight loss or a reduction in visceral fat, which could ultimately help in reducing blood pressure [12]. Participation in regular physical activity improves blood glucose control by increasing insulin sensitivity and can prevent or delay onset of type 2 diabetes [13]. Hence it is evident that regular physical activity has numerous cardio-metabolic beneficial effects and the ongoing worldwide epidemic of cardiovascular and metabolic disease is evidence to the fact that the general population is physically inactive. Results The total number of research studies included in the present review is eleven (India-8, Sri Lanka-2, Pakistan-1). In addition, country reports (n = 11, Nepal-3, Bangladesh-2, India-2, Sri Lanka-2, Bhutan-1, Maldives-1) of World Health Organization (WHO) STEPS survey for non-communicable disease risk factors for the South-Asian countries were re- trieved by searching the WHO website (http://www.who. int/chp/steps/reports/en/index.html). The sample size in research studies varied from 416–6,940 and the study setting for research studies were urban (n = 2), rural (n = 3) or both (n = 6). In the STEPS surveys sample sizes were between 2,026 and 44,491. Most research studies used self designed interviewer administered questionnaires Quality assessment adults (>15 years of age), b) Cross‐sectional study design or being the first phase of a longitudinal study, c) Geographic- ally and temporally defined population from any of the South Asian countries mentioned above, d) Being an original study presenting data on physical activity, e) Evaluating physical activity using questionnaires, f) Published in English, or with detailed summaries in English and g) Peer-reviewed fully published research papers. Studies limited to adults engaged in a particular profession, based in hospitals/institutions and confined to those with diagnosed illnesses were excluded. In addition conference proceedings, editorials, commentaries and book chapters/book reviews were excluded. All included research studies were assessed for quality and assigned a rating of either “good” (6–7 points), “fair” (4–5 points) or “poor” (1–3 points) by two reviewers (DCR and RJ). The following seven criteria were used and each one was given a point; 1) appropriateness of research design, 2) appropriate recruitment strategy, 3) adequate response rate (>50%) 4) representativeness of sample, 5) usage of objective and reliable measures to assess physical activity 6) power calculation/justification of numbers and 7) appropriateness of statistical analysis. Inclusion and exclusion criteria The following inclusion criteria were used; a) Population- based studies among healthy non-institutionalized human Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 Page 3 of 11 http://www.ijbnpa.org/content/10/1/116 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Page 3 of 11 Figure 1 Summarized search strategy. Figure 1 Summarized search strategy. Findings from research studies d h ll l In India the overall prevalence of inactivity was 18.5%- 88.4% (Table 1). In Indian males the prevalence of inactivity was 12.7%-66.2%. Majority of the studies (n = 5, 62.5%) reported a prevalence of inactivity < 23% in Indian males. In Indian females the inactivity prevalence was 17.0%-79.6%, while majority of the studies (n = 5, 62.5%) reported it to be > 39.5%. The prevalence of inactivity in urban areas of India was reported as 20.7%-88.7%, while in rural areas it was 6.6%-88.1%. One study conducted in rural India reported that physical inactivity was more at leisure time (males - 85.2%, females - 97.3%), and at work (male - 57.2%, female - 59.9%) and less during transport (male - 18.8%, female - 45.7%) [23]. A similar study in both urban and rural areas of India reported inactivity more at leisure time (74.0%) and less at work (31.0%) [24]. There were two research studies on physical activity from Sri Lanka [30,31]. One study was conducted on a nationally representative sample and the other in the most populous western province of Sri Lanka. Both studies looked at urban and rural settings and Inactivity was defined using a uniformed method (IPAQ). The prevalence of Inactivity was 11% in the national sample and 31.8% in In India the overall prevalence of inactivity was 18.5%- 88.4% (Table 1). In Indian males the prevalence of inactivity was 12.7%-66.2%. Majority of the studies (n = 5, 62.5%) reported a prevalence of inactivity < 23% in Indian males. In Indian females the inactivity prevalence was 17.0%-79.6%, while majority of the studies (n = 5, 62.5%) reported it to be > 39.5%. The prevalence of inactivity in urban areas of India was reported as 20.7%-88.7%, while in rural areas it was 6.6%-88.1%. One study conducted in rural India reported that physical inactivity was more at leisure time (males - 85.2%, females - 97.3%), and at work (male - 57.2%, female - 59.9%) and less during transport (male - 18.8%, female - 45.7%) [23]. A similar study in both urban and rural areas of India reported inactivity more at leisure time (74.0%) and less at work (31.0%) [24]. Only one STEPS survey has been carried out in Bhutan in the capital city Thimphu (2007, n = 2,484). The overall inactivity prevalence was 58.6% and females (69.6%) were more inactive than males (49.8%). Findings from STEPS surveys In the STEPS survey reports there was uniformity in the sampling and the definition of inactivity. Overall in majority of studies females were more inactive compared to males (Table 2). Furthermore, leisure related inactivity was >75% in studies reporting inactivity in this domain and people were more active in the transport domain when compared with other domains (Table 2). Two surveys were done in 2002 (n = 11,409) and 2009 (n = 9,275) in Bangladesh. In 2002, overall prevalence of physical inactivity was 52.3%. However, in this survey physical activity was categorised as light, moderate, heavy based on a likert scale; always, usually, sometimes and never. Inactivity was defined as people who have never done light/moderate activities, which was different to the criteria used in all other WHO STEPS surveys. In the national sample (2009) the overall inactivity prevalence was 27%, with more females (41.3%) being inactive than males (10.5%) (Table 2). Data extraction A separate assessment is not included for the WHO STEPS surveys conducted according to the uniform STEPS study protocol which satisfies all of the above seven criterion used for quality assessment. Definition of inactivity How sedentary lifestyle or inactivity was defined varied significantly in the different studies that used self designed questionnaires to evaluate physical activity (Table 1). Phys- ical Activity Level (PAL) (Estimated total energy expend- iture per 24 hrs / basal metabolic rate) was calculated in 3 studies and a PAL for inactivity was defined (<1.5/<1.69) [20-22]. However, there was a wide variation in the def- inition of physical inactivity in the other studies (Table 1). In the WHO STEPS surveys using the GPAQ, the total of the categorical score in the 3 domains (work, leisure time and transport) classifies an individual into three categor- ies; ‘Inactive’, ‘Moderately active’ and ‘Highly active’. The ‘Inactive’ category includes those who do not perform any physical activity or those reporting some activity, but not enough to meet other categories. The same definition was used in the IPAQ for the ‘Inactive’ category. Data extraction Data were extracted from the included studies by one reviewer using a standardised form and checked for accuracy by a second reviewer. The data extracted from each study were: a) study details (lead author, country/ city, year published/year of survey), b) methods (sample size, sampling method, age of subjects in years and type of questionnaire used, definitions), and c) physical activity data (all adults, males, females and domain specific data). Discrepancies in the extracted data were resolved by discussion, with involvement of a third reviewer when necessary. Page 4 of 11 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 the other study conducted in western province (Table 1). Males (14.6% and 38.5%) were more inactive than females (8.7% and 24.7%). Inactivity of urban adults was 35.2% and higher than rural adults (27.6%) in the study from Western province (Table 1). In the National sample; female gender (OR:2.1), age > 70 (OR:3.8), urban-living (OR:2.5), Muslim ethnicity (Sri Lankan Moor) (OR:2.7), tertiary education (OR:3.6), obesity (OR:1.8), diabetes (OR:1.6), hypertension (OR:1.2) and metabolic syndrome (OR:1.3) all significantly increased odds of being ‘inactive’ [31]. The only study on physical activity conducted in urban Pakistan using IPAQ showed that prevalence of inactivity was 60.1% (males-52.1%, females-69.8%) [29]. for evaluation of physical activity (n = 7). International Physical Activity Questionnaire (IPAQ) was used by 3 research studies and 1 study used WHO STEPS survey tool. All WHO STEPS surveys used Global Physical Activity Questionnaire (GPAQ) for the evaluation of physical activity. Majority of the research studies received a “good” (n = 5) or “fair” (n = 3) ranking on the quality assessment, while only 3 were ranked “poor” (Additional file 2). A separate assessment is not included for the WHO STEPS surveys conducted according to the uniform STEPS study protocol which satisfies all of the above seven criterion used for quality assessment. for evaluation of physical activity (n = 7). International Physical Activity Questionnaire (IPAQ) was used by 3 research studies and 1 study used WHO STEPS survey tool. All WHO STEPS surveys used Global Physical Activity Questionnaire (GPAQ) for the evaluation of physical activity. Majority of the research studies received a “good” (n = 5) or “fair” (n = 3) ranking on the quality assessment, while only 3 were ranked “poor” (Additional file 2). Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 , Rural, M 579, (Ma s, Urban uster sa 416 (M es racterist 782 (mal s Urban minister ys, carryi activities y calcula dex inclu y onnaire, zed, 1.40–1.69 onnaire, physical nactive: hour per week exercise Findings from research studies d h ll l Two large STEPS surveys were done in India in 2003–2005 (6 states, n = 44,491) and 2007–2008 (7 states, n = 39,064). In 2003 the overall prevalence of physical inactivity was 15.8% and in 2007 it was 72.3%. This variation could be attributed to different states used in the two surveys. However Kerala and Maharashtra states were evaluated in both surveys, and the overall prevalence of physical inactivity in Kerala/ Maharashtra has increased from 6.7%/6.8% in 2003 to 75.8%/81.2% in 2007. Male and Female physical inactivity in 2003 was 12.6% and 18.9% respectively. In 2007 preva- lence of Male and Female physical inactivity was 58.5% and 75.7% respectively. Overall females were more inactive than males and urban residents were more inactive than rural (Table 2). The only STEPS survey conducted in Maldives (2004) has been conducted in its’ capital city Malē (n = 2026). p g g at work, home or tran discretionary time es 3,768, Females rban, rural and ng Interviewer administe PAL was calculated an PAL <1.40 extremely in sedentary/lightly active s 260, Females 260), nd rural, Random Interviewer administe Inactive: Sedentary job exercise or cycling, M Sedentary job and som physical exercise and/o OR Standing job and n or cycling ural po at work, discretio Interview PAL was PAL <1.4 sedentary Interview Inactive: exercise Sedentar physical OR Stand or cycling Findings from research studies d h ll l More than 90% (Males 91.2%, Females 94.6%) at work, home or transport and discretionary time ●In males inactivity was 19.7%, while in females it was 17.0% 6,447, (Males 3,768, Females 7-76 yrs, Urban, rural and xed sampling Interviewer administered questionnaire, PAL was calculated and categorized, PAL <1.40 extremely inactive, PAL 1.40–1.69 sedentary/lightly active ●Extreme inactivity prevalence 9.7% (Males 7.4%, Females 12.9%), ●Sedentary/lightly active prevalence 62.1% (Males 58.8%, Females 66.7%) 520, (Males 260, Females 260), rs, Urban and rural, Random Interviewer administered questionnaire, Inactive: Sedentary job and no physical exercise or cycling, Moderately inactive: Sedentary job and some but <1 hour physical exercise and/or cycling per week OR Standing job and no physical exercise or cycling ●Prevalence of inactivity 29.4%* (Males 12.7%, Females 46.1%, Urban 29.6%, Rural 29.2%), ●Prevalence of moderate inactivity 21.5%,* (Males 25.7%, Females 17.3%, Urban 30.0%, Rural 13.1%), ●Inactivity was more in Urban and in females, ●Urban females waist circumference reduced (p < 0.05) with increased physical activity, ●BMI showed a steady decline from inactivity to activity ,825, (Males 650, Females 1175), Rural population Interviewer administered questionnaire, Sedentary lifestyle: those who had never ●40.0% had a sedentary lifestyle (Males 40.8%, Females 39.7%) males it was ales 7.4%, e prevalence .7%, Females valence of %, Females ctivity was females with ed a steady e in all s 85.2%, 8.8%, 57.2% ghest in roup 2.9%, ore than 0%), more 8.9%, n 1.64 in an n the r old not d and cised vity lev 30-1.56 > 58 yr w ove tionary ●Wo ercise les, ● n/day ross ag 6 yrs ag 5%) and w nactivity m %), and le %), ●Ina - 59.9%; ● 4 yrs) and prevalen 1.9% ), ● poration 2 ●Inactivit (31.0%), ● %, RR = 1 killed wo (32.0%, R 3%, RR = prevalenc nder diffe activity w nactivity p %), ●Sed 58.8%, F of inactiv n 29.6%, 5%) and w nactivity m %), and le %), ●Ina - 59.9%; ● 64 yrs) and prevalen 1.9% ), ● poration 2 ●Inactivit (31.0%), ● 7%, RR = 1 killed wo (32.0%, R 3%, RR = prevalenc ender diffe activity w nactivity p %), ●Sed 58.8%, F of inactiv n 29 6%, Females 39.7%) Res ●P and olde fem mal you disc cho exe 40.9 high (rec R a o f m y d c e 4 ( e, e + ure e y onnaire, zed, 1.40–1.69 onnaire, hysical nactive: hour per week exercise ose who had never minister ys, carry activities y calcula dex inclu g home or tran nary time er administer calculated an 0 extremely in /lightly active er administer Sedentary job or cycling, Mo y job and som exercise and/o ng job and n ools an ministe y level diture/E ), Seden ools an ministe y level diture/E ), Seden y y y essm rview sical a rgy ex tabolic L) <1.5 Interview Inactive: light/sede travel, Ina Composit Interview Inactive: light/sede travel, Ina Composit Sedentar at work, discretio Interview PAL was PAL <1.4 sedentar Interview Inactive: exercise Sedentar physical OR Stand or cycling 441), ent pling les 3689), atified 188) pling les 3689), atified 188) pulation s 3,768, Females rban, rural and ng 260, Females 260), d rural, Random ural po 447, (M -76 yrs ed sam 20, (Ma Urban http://www.ijbnpa.org/content/10/1/116 http://www.ijbnpa.org/content/10/1/116 (Males 52.1%, Females 69.8%) ●Females were significantly more inactive than males (OR: 2.1, 95% CI 1.5–3.1, p < 0.001) ●Prevalence of inactivity 31.8%* (Males 38.5%, Females 24.7%), ●Inactivity in urban adults 35.2% (Males 41.0%, Females 29.0), ●Inactivity in rural adults (Males 35.0%, Females 19.0%), ●Physical inactivity had a significant association with high BMI among women irrespective of their urban or rural living ●Prevalence of inactivity 11.0% (Males 14.6%, Females 8.7%) th 2% th 2% ral with % an %, 35.2% l rban or %, rban or %, an 88.7% on incre activity 6 e inactiv < 0.001) * (Males rban ad activity irrespective of thei ivity 11.0% (Males 1 n 88.7% on incre ctivity e inactiv 0.001) (Males ban ad activity ve of thei % (Males 1 4% (U perte ce of %) tly m –3.1, y 31.8 vity in 9.0), ● Exercise .1%), ●P ck of exer Males 52.1 Females ales (OR: Prevalen males 24 Males 41.0 ults (Ma BMI amon rural livin ●Prevalen Females 8 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 Page 7 of 11 http://www.ijbnpa.org/content/10/1/116 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Page 7 of 11 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Table 2 Summary of findings from STEPS surveys Country Year of study Sample characteristics Results Inactive (%) Inactivity in each domain (%) Male Female Urban Rural Total Work Transport Recreation Bangladesh 2002 Sample size 11,409 NR NR 50.1# 56.7# 52# (Male 5,625, Female 5,784) Age 25–64 yrs Urban and rural In capital city - Dhaka Bangladesh 2009-2010 Sample size 9,275 10.5 41.3 NR NR 27.0 45.7 44.5 81.9 (Male 4,312, Female 4,963) Age > 25 yrs Urban and rural Bhutan 2007 Sample size 2,484 49.8 69.6 58.6 NA 58.6 69.0 63.2 78.7 (Male 1,138, Female 1,346) Age 25–74 yrs Urban In capital city - Thimpu India 2003-2005 Sample size 44,491 (Male 21,871, Female 22,620) Age 15–64 yrs Urban and rural 6 States in India 1. Assam 8.3 10.7 26.1* 1.6* 9.5* 28.3* 24.6* NR 2. Delhi 25.5 15.5 26.3* NR 20.4* 62.9* 39.3* NR 3. Haryana 16.9 47.6 38.3* 24.6* 32.7* 78.4* 41.9* NR 4. Kerala 4.8 4.3 7.6* 5.9* 6.7* 18.4* 31.8* NR 5. Maharashtra 7.7 6.1 14.9* 1.3* 6.8* 24.5* 21.1* NR 6. http://www.ijbnpa.org/content/10/1/116 Tamil Nadu 16.9 27.2 29.1* 19.9* 22.0* 90.8* 25.1* NR India 2007-2008 Sample size 38,064 (Male 16,891, Female 21,173) Age 24–64 yrs Urban and rural 7 states in India 1. Andhra Pradesh 55.9 79.7 77.5 63.8 67.7 NR NR NR 2.Kerala 64.7 86.2 70.8 74.5 75.8 NR NR NR 3. Madhya Pradesh 33.5 52.0 68.3 31.8 42.3 NR NR NR 4. Maharashtra 75.4 87.7 86.1 77.2 81.2 NR NR NR 5. Mizoram 60.9 82.4 79.1 62.5 71.1 NR NR NR 6. Tamil Nadu 57.3 74.2 79.4 61.6 65.8 NR NR NR 7. Uttarakhand 64.6 69.7 91.6 57.6 67.1 NR NR NR Maldives 2004 Sample size 2,026 NR NR NR NA NR 93.2* NR NR (Male 934, Female 1,092) Age 25–64 yrs Urban (In Male) Table 2 Summary of findings from STEPS surveys Country Year of study Sample characteristics Ranasinghe et al. 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International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Page 8 of 11 Table 2 Summary of findings from STEPS surveys (Continued) Nepal 2003 Sample size 2,030 73.6 90.9 NR NA 82.3* 82.3* 27.6* 94.5* (Male 1,010, Female 1,020) Age 25–64 yrs Urban Nepal 2004-2005 Sample size 7,792 NR NR NR NR NR 51.5 19.1 86.0 (Male 3,674, Female 4,118) Age 15–64 yrs Urban and rural Nepal 2007 Sample size 4,328 5.2 5.9 NR NR 5.5 10.6 19.0 83.2 (Male 1,907, Female 2,421) Age 15–64 yrs Urban and rural 15 of 75 districts Sri Lanka 2003 Sample size 3,000 12.1 19.1 NR NR 15.6 58.3* NR 94.8* (Male 1,500, Female 1,500) Age 15–74 yrs Urban One of nine provinces - Western Sri Lanka 2006 Sample size 11,680 17.9 31.9 NR NR 25.0 NR NR NR (Male 5,765, Female 5,915) Age 24–64 yrs Urban and rural 5 random districts out of all 25 districts * - calculated from available data; NR – Not reported; NA – Not applicable. http://www.ijbnpa.org/content/10/1/116 Table 2 Summary of findings from STEPS surveys (Continued) - calculated from available data; NR – Not reported; NA – Not applic of the respondents reported that they are physically inactive at work. In contrast STEPS survey data on physical activity were available from all countries except Pakistan. All research studies from the region collectively evaluated physical activity of 26,360 adults, while STEPS surveys contained data on 136,579 adults. We observed a marked variation between research studies in the definition of physical inactivity and in the questionnaires used. Hence, comparisons between studies were undertaken with caution. This barrier was overcome in the STEPS surveys as they used the GPAQ to evaluate physical activity. It is important that future researchers try and adhere to this uniformity in order to derive intra- and inter-regional comparable data and observe secular trends in physical activity. The present study mainly focuses on the preva- lence of physical inactivity and its variations with gender and area of residence. We also tried to identify inactivity levels in different domains (work, transport, leisure) as occupations, transport methods, social and cultural values differ in this region compared to other regions and the developed world. Three STEPS surveys have been conducted in Nepal. In 2003 only an urban sample was taken from the capital city (n = 2,030). In later STEPS surveys both urban and rural samples were studied (2004 n = 7,792 and 2007 n = 4,328). The 2003 survey reported an overall inactivity of 82.9% (Males 74.6%, Females 91.2%) in the urban residents. However in contrast the 2007 national sample (conducted in 15 out of 75 districts) reported that overall inactivity was only 5.5% (Males 5.2%, Females 5.9%) (Table 2). In Sri Lanka two STEPS surveys were done in 2003 (single province, n = 3,000) and 2006 (five districts, n = 11,680). In 2003 overall prevalence of physical inactivity was 15.6% and in 2006 it has increased to 25%. Females (2003–19.1%, 2006–31.9%) were more inactive than males (2003–12.1%, 2006–17.9%) in both studies. Discussion To our knowledge this is the first systematic review evaluating physical inactivity and physical activity patterns among adults from South Asian countries. Published research articles evaluating physical activity prevalence were only available from India, Sri Lanka and Pakistan. We observed several socio-economical factors associated with physical inactivity. Skilled workers and professionals were more inactive than unskilled workers in the region [24]. Similarly, higher education was a significant factor Page 9 of 11 associated with physical inactivity [31]. Among South Indians, Vaz et al. reported that people engaging in rec- reational exercise were inactive in other domains [20,21]. Gender is also an important factor to determining physical activity levels of a population. Several studies reported higher physical inactivity among South Asian women [23,25,29,31,32]. Cultural expectations may restrict the participation of women in certain forms of physical activity in some religious and ethnic groups in the region. The traditional role of South Asian women in taking care of household work and supporting extended family members may limit the time available for them to engage in physical activity, in particularly leisure time physical activities. Low physical activity is one of the contributing risk factors for the higher obesity levels seen among Asian Indians [33]. A qualitative study conducted among South Asians living in United Kingdom found that understanding external motivators and social context of their lives is very crucial for developing successful physical activity interventions [34]. Hence, although exercise is promoted in public health campaigns to increase the overall physical activity level of a population, it is important to under- stand socio-economical factors associated with different populations in order to deliver effective physical activity interventions. which were studied in both surveys also showed marked variation. This could be due to differences in the sampling frames used. However such dramatic increases in inactiv- ity over a short period warrants further investigation. The only surveys conducted in urban capitals of Maldives and Bhutan had significantly higher prevalence of inactivity. In Bhutan inactivity was lower compared to Maldives due to it being mostly an agrarian society with nearly 50% of the labour force engaged in agriculture as compared to Maldives where most are involved in the tourism industry [37]. The most active regional country in the STEPS surveys was Nepal, possibly due to 80% of the Nepalese population being involved in agriculture [38]. Discussion In all regional countries work and transport related activity was higher compared to leisure time activity. Hence it is important to consider individual country profiles, proportions of urbanity and cultures when delivering public health mes- sages focused on leisure time activity. Comparable nationally representative country data are available from the WHO World Health Survey (IPAQ questionnaire) conducted in 2002–2003, where the male and female inactivity prevalence was as follows; India 9.3% & 15.2%, Nepal 6.7% & 9.7% and Sri Lanka 7.3% & 13.8% [39]. The data are fairly comparable to those from WHO STEPS surveys in India (2003) and Nepal (2007). In contrast in Sri Lanka inactivity has markedly increased from the 2002 World Health Survey to the 2006 STEPS survey. Several studies show that IPAQ overestimates inactivity prevalence and under estimates prevalence in rural areas of developing countries [32,40,41]. The STEPS surveys have also been conducted in 80 developing countries around the world allowing for a meaningful comparison across regions [35]. In the Western Pacific region (19 countries) inactivity prevalence ranged from 7.5% in Mongolia to 75.3% in Cook Islands with majority of countries falling between 40%-66%. Eastern Mediterranean (14 countries) prevalence of inactivity ranged from 30%-60%. In Africa (31 countries) lowest prevalence of inactivity was observed in Mozambique (6.4%) and highest in Zimbabwe (79.3%). In nearly half of the African countries inactivity was <29%. East Asian countries like Myanmar (12.7%) and Indonesia (22.6%) had prevalence’s of inactivity comparable with the South Asia and Africans. With the available data we observe that the persons residing in South Asian region are more active compared with the Western Pacific and Mediterranean countries, lying in parallel with East Asia and Africa. In developing countries with agrarian economies inactivity was low irrespective of the region (Nepal, Mongolia, Malawi, Mozambique and Cambodia). However the inactivity prevalence seems to be on the rise in most Western Pacific countries due to recent economic transitions. Hence, countries in the South Asian region in economic transition also need public health planning to enable the people to maintain their already existing active The WHO STEPwise approach was introduced for chronic disease risk factor surveillance in order to close the gap in the availability of data from developing countries and to strengthen their capacity to conduct such surveillances [35]. The GPAQ used in these surveys not only gave uniformity to the gathered data, it also allowed meaningful comparisons. Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Discussion The GPAQ is known to be reliable for surveys in developing countries, adaptable to incorporate cultural differences [36]. The initial STEPS surveys from the region were mostly in sub-populations and confined to logistically favourable areas like capitals. Sri Lanka (2006), Nepal (2007) and Bangladesh (2009) (Inactivity: 25%, 5.5% and 27% respectively) had nationally representative samples in subsequent surveys. Prevalence of overall inactivity reduced in these subsequent surveys as the sample dispersed from populations in urban capitals to the rural parts of the countries. This highlights the effect of urbanity on physical activity. However, in the Sri Lankan STEPS surveys inactivity prevalence has increased in the second survey (2006), which used a national sample. This is probably due to the initial survey (2003) being conducted in more rural areas of the Western province of Sri Lanka, in contrast to Nepal and Bangladesh where the initial surveys were from more urbanised capitals. The two large STPES surveys (2003, 2007) from India contrasted in the prevalence of inactivity, possibly attribut- able to subjective variations in data collection, inter-state variations and differences in time and season of data col- lection. However, data from Kerala and Maharashtra states Page 10 of 11 life styles. We also observe that females were more inactive in the South Asian region when compared to males, a finding which is seen in most other regions and even in developed countries. leisure time. These factors need to be considered when planning future interventions and research aimed at improving PA in the region. However, future researchers should try and adhere to uniformity in definitions and assessment tools in order to derive intra- and inter-regional comparable data and observe secular trends. The International Prevalence Study conducted in 2002– 2004 using IPAQ categorized inactivity to be least in China (6.9%), New Zealand (12.2%), Canada (13.7%), USA (15.9%) and Australia (17.2%) [42]. We observe that South Asian activity levels were in parallel or sometimes better than these developed countries which have longer physical activity promotion strategies, mainly targeted at improving leisure time physical activity. However with increased urbanisation and busy work schedules the leisure time is also reduced. In the South Asian region we observed that activity during work and transport is higher than leisure time activity. Promoting leisure time activity has also become a challenge in South Asia due to cultural and attitudinal barriers. Discussion Hence public health messages in the region should be directed at improving all domains of activity with work and transport policies of the countries supporting them. Competing interests The authors declare that they have no competing interests. Received: 7 April 2013 Accepted: 9 October 2013 Published: 12 October 2013 Received: 7 April 2013 Accepted: 9 October 2013 Published: 12 October 2013 References 1. Gupta M, Singh N, Verma S: South Asians and cardiovascular risk: What clinicians should know. Circulation 2006, 113(25):e924–e929. 1. Gupta M, Singh N, Verma S: South Asians and cardiovascular risk: What clinicians should know. Circulation 2006, 113(25):e924–e929. 2. Eapen D, Kalra GL, Merchant N, Arora A, Khan BV: Metabolic syndrome and cardiovascular disease in South Asians. Vasc Health Risk Manag 2009, 5:731–743. 2. Eapen D, Kalra GL, Merchant N, Arora A, Khan BV: Metabolic syndrome and cardiovascular disease in South Asians. Vasc Health Risk Manag 2009, 5:731–743. 3. Erens B, Primatesta P, Prior G: The health of minority ethnic groups ′99 methodology & documentation. London: The health survey for england; 2001. 4 Jayawardena R Ranasinghe P Byrne NM Soares MJ Katulanda P Hills AP: 3. Erens B, Primatesta P, Prior G: The health of minority ethnic groups ′99 methodology & documentation. London: The health survey for england; 2001. 4. Jayawardena R, Ranasinghe P, Byrne NM, Soares MJ, Katulanda P, Hills AP: Prevalence and trends of the diabetes epidemic in South Asia: a systematic review and meta-analysis. BMC Public Health 2012, 12:380. 5. Gupta M, Brister S: Is South Asian ethnicity an independent cardiovascular risk factor? Can J Cardiol 2006 22(3):193 197 4. Jayawardena R, Ranasinghe P, Byrne NM, Soares MJ, Katulanda P, Hills AP: Prevalence and trends of the diabetes epidemic in South Asia: a systematic review and meta-analysis. BMC Public Health 2012, 12:380. 5. Gupta M, Brister S: Is South Asian ethnicity an independent cardiovascular risk factor? Can J Cardiol 2006, 22(3):193–197. 6. Qin L, Corpeleijn E, Jiang C, Thomas GN, Schooling CM, Zhang W, Cheng KK, Leung GM, Stolk RP, Lam TH: Physical activity, adiposity, and diabetes risk in middle-aged and older Chinese population: The Guangzhou Biobank Cohort Study. Diab Care 2010, 33(11):2342–2348. Authors’ contributions One of the strengths of the current systematic review is the comprehensive and easily replicable search strategy applied to 3 major medical databases. In addition we systematically selected the studies through the application of well-defined inclusion/exclusion criteria. We would also like to highlight several limitations in the present review. Firstly, most of studies were limited to regional populations. As South Asian countries have considerably diverse ethnic groups, the regional findings may not be generalized to whole country or for the entire South Asian region. Secondly, there is no uniformity on physical activity assessment tools. Although several of studies have used IPAQ, differences in the formats used limit comparability. For instance, Arambepola et al. has used longer version of IPAQ whereas Katulanda et al. has used a short version [30,31]. Thirdly, there is clear heterogeneity on the defin- ition of physical inactivity/sedentary life style and physical activity levels. Furthermore, only limited number of articles reported data on the sub-domains of physical activity (work, transport and leisure). In addition, although many studies have shown that resistance exercise improves over- all health and that it is beneficial for South Asians with Diabetes at present there is only very limited data available for South Asians [43,44]. DC, PR and RJ made substantial contribution to conception and study design. DC, PR and RJ were involved in data collection. DC, PR, RJ and AM were involved in refining the study design, statistical analysis and drafting the manuscript. DC, PR, RJ and AM critically revised the manuscript. All authors read and approved the final manuscript. Additional files Additional file 1: PRISMA 2009 checklist. Additional file 2: Assessment of Quality of the included Research Studies. Additional file 1: PRISMA 2009 checklist. Additional file 2: Assessment of Quality of the included Research Studies. Abbreviations Abbreviations GPAQ: Global physical activity questionnaire; IPAQ: International physical activity questionnaire; MeSH: Medical subject heading; MET: Metabolic equivalents; OR: Odds ratio; PAL: Physical activity level; PRISMA: Preferred reporting in systematic reviews & Meta-Analysis; WHO: World Health Organization. Author details 1 1Allied Health Sciences Unit, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka. 2Department of Pharmacology, Faculty of Medicine, University of Colombo, Colombo, Sri Lanka. 3Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, Australia. 4Fortis-C-DOC Centre of Excellence for Diabetes, Metabolic Diseases and Endocrinology, Chirag Enclave, New Delhi, India. Received: 7 April 2013 Accepted: 9 October 2013 Published: 12 October 2013 Conclusions Jepson R, Harris FM, Bowes A, Robertson R, Avan G, Sheikh A: Physical activity in South Asians: An in-depth qualitative study to explore motivations and facilitators. PLoS One 2012, 7(10):e45333. 12. Reaven PD, Barrett-Connor E, Edelstein S: Relation between leisure-time physical activity and blood pressure in older women. Circulation 1991, 83(2):559–565. 35. 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In the South Asian regions females, skilled workers, professionals and those with higher education were more inactive. Majority of South Asian adults were inactive during their 8. Aust NZJ, MedCaspersen CJ, Powell KE, Christenson GM: Physical activity, exercise, and physical fitness: Definitions and distinctions for health-related research. Public Health Rep 1985, 100(2):126–131. 9. Willett WC, Koplan JP, Nugent R, Dusenbury C, Puska P, Gaziano TA: 8. Aust NZJ, MedCaspersen CJ, Powell KE, Christenson GM: Physical activity, exercise, and physical fitness: Definitions and distinctions for health-related research. Public Health Rep 1985, 100(2):126–131. 9. Willett WC, Koplan JP, Nugent R, Dusenbury C, Puska P, Gaziano TA: Prevention of chronic disease by means of diet and lifestyle changes. In Disease control priorities in developing countries. 2nd edition. Edited by 9. Willett WC, Koplan JP, Nugent R, Dusenbury C, Puska P, Gaziano TA: Prevention of chronic disease by means of diet and lifestyle changes. In Disease control priorities in developing countries. 2nd edition. Edited by Page 11 of 11 Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Jamison DT, Breman JG, Measham AR, Alleyne G, Claeson M, Evans DB, Jha P, Mills A. Washington (DC): Musgrove P; 2006. Jamison DT, Breman JG, Measham AR, Alleyne G, Claeson M, Evans DB, Jha P, Mills A. Washington (DC): Musgrove P; 2006. 32. Ekelund U, Sepp H, Barge S: Criterion-related validity of the last 7-day, short form of the International Physical Activity questionnaire in Swedish adults. Public Health Nutr 2006, 9:258–265. 10. Fox KR: The influence of physical activity on mental well-being. Public Health Nutr 1999, 2(3A):411–418. 33. Chopra SM, Misra A, Gulati S, Gupta R: Overweight, obesity and related non-communicable diseases in Asian Indian girls and women. Eur J Clin Nutr 2013, 67(7):688–696. 11. Ishikawa K, Ohta T, Zhang J, Hashimoto S, Tanaka H: Influence of age and gender on exercise training-induced blood pressure reduction in systemic hypertension. Am J Cardiol 1999, 84(2):192–196. 34. Conclusions 19. Moher D, Liberati A, Tetzlaff J, Altman DG, Group P: Preferred reporting items for systematic reviews and meta-analyses: The PRISMA statement. BMJ 2009, 339:b2535. 43. Hills AP, Shultz SP, Soares MJ, Byrne NM, Hunter GR, King NA, Misra A: Resistance training for obese, type 2 diabetic adults: a review of the evidence. Obes Rev 2010, 11(10):740–749. 20. Vaz M, Bharathi AV, Kurpad AV: Exercising’ but not active: Implications for physical activity counselling. Natl Med J India 2006, 19(6):345. 44. Misra A, Alappan NK, Vikram NK, Goel K, Gupta N, Mittal K, Bhatt S, Luthra K: Effect of supervised progressive resistance-exercise training protocol on insulin sensitivity, glycemia, lipids, and body composition in Asian Indians with type 2 diabetes. Diabetes Care 2008, 31(7):1282–1287. 21. Vaz M, Bharathi AV: Perceptions of the intensity of specific physical activities in Bangalore, South India: Implications for exercise prescription. J Assoc Physicians India 2004, 52:541–544. 22. Sullivan R, Kinra S, Ekelund U, Bharathi AV, Vaz M, Kurpad A, Collier T, Reddy KS, Prabhakaran D, Ben-Shlomo Y, et al: Socio-demographic patterning of physical activity across migrant groups in India: results from the Indian Migration Study. PLoS One 2011, 6(10):e24898. doi:10.1186/1479-5868-10-116 Cite this article as: Ranasinghe et al.: Physical activity patterns among South-Asian adults: a systematic review. International Journal of Behavioral Nutrition and Physical Activity 2013 10:116. 23. Krishnan A, Shah B, Lal V, Shukla DK, Paul E, Kapoor SK: Prevalence of risk factors for non-communicable disease in a rural area of Faridabad district of Haryana. Indian J Public Health 2008, 52(3):117–124. y 24. Sugathan TN, Soman CR, Sankaranarayanan K: Behavioural risk factors for 24. Sugathan TN, Soman CR, Sankaranarayanan K: Behavioural risk factors for non communicable diseases among adults in Kerala, India. Indian J Med Res 2008, 127(6):555–563. non communicable diseases among adults in Kerala, India. Indian J Med Res 2008, 127(6):555–563. 25. Agrawal VK, Basannar DR, Sing RP, Dutt M, Abraham D, Mustafa MS: Coronary risk factors in a rural community. Indian J Public Health 2006, 50(1):19–23. g g Coronary risk factors in a rural community. Indian J Public Health 2006, 50(1):19–23. 26. Mittal M, Arora M, Bachhel R, Kaur N, Sidhu RS: Physical activity, indices of obesity and mean arterial blood pressure: Does place of living matters? rural vs urban. J Clin Diagn Res 2011, 5(5):1038–1042. 27. Haldiya KR, Mathur ML, Sachdev R: Lifestyle-related risk factors for cardiovascular disease in a desert population of India. Ranasinghe et al. International Journal of Behavioral Nutrition and Physical Activity 2013, 10:116 http://www.ijbnpa.org/content/10/1/116 Conclusions Curr Sci 2010, 99(2):190–195. 28. Agrawal R, Chaturvedi M, Singh S, Gupta SC: An epidemiological study of dietary and exercise habits as correlates of hypertension in persons aged 45 years and above in Agra District. Indian J Comm Health 2012, 24(2):91–96. Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: • Convenient online submission • Thorough peer review • No space constraints or color ﬁgure charges • Immediate publication on acceptance • Inclusion in PubMed, CAS, Scopus and Google Scholar • Research which is freely available for redistribution Submit your manuscript at www.biomedcentral.com/submit Submit your next manuscript to BioMed Central and take full advantage of: 29. Khuwaja AK, Kadir MM: Gender differences and clustering pattern of behavioural risk factors for chronic non-communicable diseases: Community-based study from a developing country. Chronic Illn 2010, 6(3):163–170. • Convenient online submission 30. Arambepola C, Allender S, Ekanayake R, Fernando D: Urban living and obesity: Is it independent of its population and lifestyle characteristics? Trop Med Int Health 2008, 13(4):448–457. 31. Katulanda P, Jayawardana R, Ranasinghe P, Rezvi Sheriff M, Matthews DR: Physical activity patterns and correlates among adults from a developing country: The Sri Lanka Diabetes and Cardiovascular Study. Public Health Nutr 2013, 16(9):1684–1692. 31. Katulanda P, Jayawardana R, Ranasinghe P, Rezvi Sheriff M, Matthews DR: Physical activity patterns and correlates among adults from a developing country: The Sri Lanka Diabetes and Cardiovascular Study. Public Health Nutr 2013, 16(9):1684–1692.
https://openalex.org/W1984632802	https://zenodo.org/records/2137248/files/article.pdf	English	null	The Bureau of Government Laboratories for the Philippine Islands, and Scientific Positions Under It	Science	1,902	public-domain	1,258	SCIENCE. SCIENCE. 579 OCTOBER 10, 1902.1 thus enabled to have his apparatus and ap- pliances in the most convenient form. The division of the space is as follows: thus enabled to have his apparatus and ap- pliances in the most convenient form. The division of the space is as follows: era1 special cryogenic laboratories have been established for the prosecution of such researches, and a liquid-air plant is becoming a common adjunct to the equip- ment of the ordinary laboratory. GROUND FLOOR. Physical laboratory with constant temperature room below. GROUND FLOOR. Physical laboratory with constant tem room below. Assay laboratory. Balance room. Combustion room. Distilling room. Research room products. for ~~egetable Chemical stores. Apparatus stores. Storekeeper's office: Bacteriological diagnosis, two rooms. Animal parasites. Culture media. Mechanic. Incubator and cold storage. JAMES DEWAR Assay laboratory. (To be concluded.) THE BUREAU OF GOVERNMENT LABORATO- RIES FOR THE PHILIPPINE ISLANDS, AND SCIENTIFIC POSITIONS UNDER IT. THE Bureau of Government Laboratories for the Philippine Islands has now been organized for nine months and is at present quartered in a temporary building. The commission contemplates the erection of a comprehensive and fitting structure for sci- entific work, the detailed plans of which are about completed, and the government archi- tect is ready to begin work as soon as the title to the land desired for the edifice is secured. This new structure will be fitted with all modern appliances for thorough scientific work. The individual working desks of the laboratory will be supplied with gas, water, vacuum and steam and air pressure; electric power is to be fnrnished wherever it is needed, and the equipment will be complete. FIRST FLOOR. Mineral analysis. Director chemical laboratory. Director's office. Sugar and food analysis. Library. Plant pathology. Biological director's office. Biological director's laboratory. Biological research. Spectroscopic rooms. Chemical research. Pharmacology. Balance room. Photography. , . Collections. Pathology. Physiological chemist. Three research rooms. Outdoor laboratory. FIRST FLOOR. Mineral analysis. Director chemical laboratory. Director's office. Sugar and food analysis. Library. Plant pathology. Biological director's office. Biological director's laboratory. Biological research. Spectroscopic rooms. Chemical research. Pharmacology. Balance room. Photography. , . Collections. Pathology. Physiological chemist. Three research rooms. Outdoor laboratory. SCIENCE. The scheme of the bureau contemplates a central institution in which laboratory work shall be done for all the bureaus which may need scientific assistance, so that a scattering of individual laboratories and a consequent loss of efficiency and equip- ment are avoided. The work is separated into two divisions, the chemical laboratory and the biological laboratory, each occupy- ing a wing of the new building, with the Serum Institute located to the rear, in con- junction with the power house. POWER HOUSE. Cold-storage plant and cold-storage rooms. Serum packing room. 6 Serum laboratory. Serum laboratory. Serum kitchen. Crematory. Engine room. Boiler room. The plans of the bureau contemplate re- search work not only in the resources of the islands, but also in the realm of tropical diseases. The work during the past year has included a large number of analyses for The building is divided into sixty rooms so that separate lines of work can be car- ried on in individual quarters, each person engaged in saientific investigation being SCIENCE. 580 [N. S. VOL. XVI. NO.406. the Custorn House, Mining .Bureau, Forest- ry Bureau, Agricultural Department and Board of Health; diagnostic work for the hospitals and others interested, and re- searches in gutta-percha, rubber and gums found in the islands, as well as investiga- tions of some previously unknown forms of tropical diseases. The scope of the work is continually widening, and there is no doubt but that the bureau offers large op- portunities for young men who desire to acquaint themselves with. the products of the tropics and to advance our knowledge of lines of work which are each year con- centrating more and more of the interest of the scientific world. the Custorn House, Mining .Bureau, Forest- ry Bureau, Agricultural Department and Board of Health; diagnostic work for the hospitals and others interested, and re- searches in gutta-percha, rubber and gums found in the islands, as well as investiga- tions of some previously unknown forms of tropical diseases. The scope of the work is continually widening, and there is no doubt but that the bureau offers large op- portunities for young men who desire to acquaint themselves with. the products of the tropics and to advance our knowledge of lines of work which are each year con- centrating more and more of the interest of the scientific world. sitions in the United States. It is the in- tention to engage none but the most efficient workers in the corps, and it is hoped, in the course of a few years, a connection with the Bureau of Government Laboratories will be equivalent to a certificate of the su- perior attainments. The plan of the institution contemplates the reservation of a certain number of re- search rooms in the laboratory building. POWER HOUSE. These are to be at the disposition of inde- pendent investigators who wish to come to the islands for a temporary period as the guests of the laboratories. These workers will be furnished all the laboratory facili- ties they desire, and it is hoped that the opportunities offered will render scientific study in the tropics easy of access to all who have planned to undertake certain lines of work in which they are interested. The positions in the bureau, outside of the directors, are all under the Civil Ser- vice, and qualifications can be obtained through the Civil Service Commission at MTashington. The scheme of the bureau contemplates the following additions to the laboratory force during the next year: - PAULC. FREER Superil~tendent of Governrizent I;nborntories. Superil~tendent of Governrizent I;nborntories. Superil~tendent of Governrizent I;nborntories. THE CARNEGIE INSTITUTION. 1 Soil and water analyst.. .......... $1,500 1 Plant pathologist.. ................ 2,500 1 Physical chemist.. . . . . . . . . . . . . . . . . 2,400 1 Analytical chemist for mineral analysis ....................... 2,000 1 Assayer ......................... 1,500 1 Entomologist ..................... 2,500 1 Animal parasitologist.. - ............. 2,500 1 Pathologist ...................... 2,400 1 Drug assayer and toxicologist.. .... 1,800 AID to research may be given either to individuals or to groups or organizations of individuals. One of the chief obstacles in this country at present to research by individuals is the lack of time for continuous, well-adjusted work. The majority of the persons engaged in active scientific investigation in the United States are connected with colleges or universities, and in nearly every instance definite accomplishmeilt is expected from them in the way of instruction and admin- istration. The exigencies-real or fancied -of university administration often lead to wasteful repetition of courses and to the exhaustion of energy in barren details of executive routine and elementary instruc- tion. The most common complaint heard from American men of science is not re- garding inadequate salaries, but regarding the scanty time afforded them for the work of investigation. While in some cases this The candidates for the higher salaried positions by understanding will not be sub- jected to a rigid examination, but their pre- vious research work, experience, university degrees and general lrnowledge will qualify them, after the facts have been submitted to the Civil Service Board and found satis- factory. The salaries for, young men are good, and, although expenses in Manila are higher than in the United States, nevertheless, the difference in salaries is large enough so that prospective workers will be better paid here than they would in the beginning po-
https://openalex.org/W4361241853	https://aacr.figshare.com/articles/journal_contribution/Supplementary_Figure_1_from_Tumor_Detection_by_Imaging_Proteolytic_Activity/22386581/1/files/39832049.pdf	English	null	Supplementary Figure 1 from Tumor Detection by Imaging Proteolytic Activity	null	2,023	cc-by	88	Supplementary Methods: Fluorescent Microscopy Image Correction Supplementary Methods: Fluorescent Microscopy Image Correction Figure 1: Images were collected using a Zeiss Axiocam and Axiovision Software and saved as TIFF files. To correct for dust/condensation on the camera, the image was collected under identical conditions with a sample of PBS only (Figure 1A). The aberrations were selected using Adobe Photoshop. This layer was then copied to cell images (1B) and filled with the appropriate color and Gaussian blurred to obtain final results (1C). + A B C + A A
https://openalex.org/W1565729601	https://ora.ox.ac.uk/objects/uuid:c0293da1-157d-4a96-8ab6-20f64e3e6260/files/mc061ba854dc071bab4d47bfd53379420	English	null	Facility location with double-peaked preferences	Autonomous agents and multi-agent systems	2,017	cc-by	19,818	Aris Filos-Ratsikas and Jie Zhang were supported by the ERC Advanced Grant 321171 (ALGAME) and the Sino-Danish Center for the Theory of Interactive Computation, funded by the Danish National Research Foundation and the National Science Foundation of China (under the Grant 61061130540), and by the Center for research in the Foundations of Electronic Markets (CFEM), supported by the Danish Strategic Research Council. Minming Li was partly supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China [Project No. CityU 117913]. Qiang Zhang was supported by FET IP Project MULTIPEX 317532. Auton Agent Multi-Agent Syst (2017) 31:1209–1235 DOI 10.1007/s10458-017-9361-0 B Aris Filos-Ratsikas Aris.Filos-Ratsikas@cs.ox.ac.uk Facility location with double-peaked preferences Aris Filos-Ratsikas1 · Minming Li2 · Jie Zhang3 · Qiang Zhang4 Published online: 20 March 2017 © The Author(s) 2017. This article is an open access publication Published online: 20 March 2017 © The Author(s) 2017. This article is an open access publication Published online: 20 March 2017 © The Author(s) 2017. This article is an open access publication Published online: 20 March 2017 Abstract We study the problem of locating a single facility on a real line based on the reports of self-interested agents, when agents have double-peaked preferences, with the peaks being on opposite sides of their locations. We observe that double-peaked preferences capture real- life scenarios and thus complement the well-studied notion of single-peaked preferences. As a motivating example, assume that the government plans to build a primary school along a street; an agent with single-peaked preferences would prefer having the school built exactly next to her house. However, while that would make it very easy for her children to go to school, it would also introduce several problems, such as noise or parking congestion in the morning. A 5-min walking distance would be sufﬁciently far for such problems to no longer be much of a factor and at the same time sufﬁciently close for the school to be easily accessible by the Aris Filos-Ratsikas and Jie Zhang were supported by the ERC Advanced Grant 321171 (ALGAME) and the Sino-Danish Center for the Theory of Interactive Computation, funded by the Danish National Research Foundation and the National Science Foundation of China (under the Grant 61061130540), and by the Center for research in the Foundations of Electronic Markets (CFEM), supported by the Danish Strategic Research Council. Minming Li was partly supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China [Project No. CityU 117913]. Qiang Zhang was supported by FET IP Project MULTIPEX 317532. B Aris Filos-Ratsikas Aris.Filos-Ratsikas@cs.ox.ac.uk Minming Li minming.li@cityu.edu.hk Jie Zhang jie.zhang@soton.ac.uk Qiang Zhang csqzhang@gmail.com 1 University of Oxford, Oxford, UK 2 City University of Hong Kong, Kowloon, Hong Kong 3 University of Southampton, Southampton, UK 4 Sapienza University of Rome, Rome, Italy 123 1 University of Oxford, Oxford, UK 123 1210 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 children on foot. There are two positions (symmetrically) in each direction and those would be the agent’s two peaks of her double-peaked preference. Motivated by natural scenarios like the one described above, we mainly focus on the case where peaks are equidistant from the agents’ locations and discuss how our results extend to more general settings. We show that most of the results for single-peaked preferences do not directly apply to this setting, which makes the problem more challenging. As our main contribution, we present a simple truthful- in-expectation mechanism that achieves an approximation ratio of 1+b/c for both the social and the maximum cost, where b is the distance of the agent from the peak and c is the minimum cost of an agent. For the latter case, we provide a 3/2 lower bound on the approximation ratio of any truthful-in-expectation mechanism. We also study deterministic mechanisms under some natural conditions, proving lower bounds and approximation guarantees. We prove that among a large class of reasonable strategyproof mechanisms, there is no deterministic mechanism that outperforms our truthful-in-expectation mechanism. In order to obtain this result, we ﬁrst characterize mechanisms for two agents that satisfy two simple properties; we use the same characterization to prove that no mechanism in this class can be group- strategyproof. Keywords Facility location · Strategyproofness · Double-peaked preferences · Approximate mechanism design without money · Social cost · Maximum cost 123 1 Introduction For that reason, following the literature we assume that 123 1211 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 the cost functions are the same for all agents and that the cost increases linearly, at the same rate, as the output moves away from the peaks. The straightforward extension to the double- peaked case is piecewise-linear cost functions, with the same slope in all intervals, which gives rise to the natural model of symmetric agents, i.e. the peaks are equidistant from the agent’s location. Note that this symmetry is completely analogous to the single-peaked case (for facility location problems, e.g. see [30]), where agents have exactly the same cost on two points equidistant from their peaks. Our lower bounds and impossibility results naturally extend to non-symmetric settings, but some of our mechanisms do not. We discuss those extensions in Sect. 6. the cost functions are the same for all agents and that the cost increases linearly, at the same rate, as the output moves away from the peaks. The straightforward extension to the double- peaked case is piecewise-linear cost functions, with the same slope in all intervals, which gives rise to the natural model of symmetric agents, i.e. the peaks are equidistant from the agent’s location. Note that this symmetry is completely analogous to the single-peaked case (for facility location problems, e.g. see [30]), where agents have exactly the same cost on two points equidistant from their peaks. Our lower bounds and impossibility results naturally extend to non-symmetric settings, but some of our mechanisms do not. We discuss those extensions in Sect. 6. Our model also applies to more general spaces, beyond the real line. One can imagine for instance that the goal is to build a facility on the plane where for the same reasons, agents would like the facility to be built at some distance from their location, in every direction. This translates to an agent having inﬁnitely many peaks, located on a circle centered around her location. In that case of course, we would no longer refer to agents’ preferences as double- peaked but the underyling idea is similar to the one presented in this paper. We do not explore such extensions here; we leave that for future work. Agents are self-interested entities that wish to minimize their costs. 1 Introduction We study the problem of locating a single facility on a real line, based on the input provided by selﬁsh agents who wish to minimize their costs. Each agent has a location xi ∈R which is her private information and is asked to report it to some central authority, which then decides where to locate the facility, aiming to optimize some function of the agents’ reported locations. This model corresponds to problems such as ﬁnding the ideal location for building a primary school or a bus stop along a street, so that the total distance of all agents’ houses from the location is minimized, or so that no agent’s house will lie too far away from that location. In our setting, we assume that agents have double-peaked preferences, i.e. we assume that each agent i has two unique most preferred points or peaks, located at some distances from xi on opposite sides, where her cost is minimum. Traditionally, preferences in facility location problems are assumed to be single-peaked, i.e. each agent’s location is her most preferred point on the line and furthermore the cost is assumed to be linear, i.e. it increases linearly (at the same rate) to the left and the right of that peak. Sometimes however, single- peaked preferences do not model real-life scenarios accurately. Take for instance the example mentioned above, where the government plans to build a primary school on a street. An agent with single-peaked preferences would deﬁnitely want the school built next to her house, so that she wouldn’t have to drive her children there everyday. However, it is quite possible that she is also not very keen on the inevitable drawbacks of having a primary school next to her house either, like unpleasant noise or trouble with parking. On the other hand, a 5- min walking distance is sufﬁciently far for those problems to no longer be a factor but also sufﬁciently close for her children to be able to walk to school. There are two such positions, (symmetrically) in each direction, and those would be her two peaks. Our primary objective is to explore double-peaked preferences in facility location settings similar to the ones studied extensively for single-peaked preferences throughout the years [1,11,14,18,20,23,26,27,30,32]. 1 Introduction We are interested in mechanisms that ensure that agents are not incentivized to report anything but their actual locations, namely strategyproof mechanisms. We are also interested in group strategyproof mechanisms, i.e. mechanisms that are resistant to manipulation by coalitions of agents. More- over, we want those mechanisms to achieve some good performance guarantees, with respect to our goals. If our objective is to minimize the sum of the agent’s costs, known as the social cost, then we are looking for strategyproof mechanisms that achieve a social cost as close as possible to that of the optimal mechanism, which need not be strategyproof. The prominent measure of performance for mechanisms in computer science literature is the approximation ratio [2,6,12,24], i.e. the worst possible ratio of the social cost achieved by the mechanism over the minimum social cost over all instances of the problem. The same holds if our objec- tive is to minimize the maximum cost of any agent. In the case of randomized mechanisms, i.e. mechanisms that output a probability distribution over points in R, instead of a single point, as a weaker strategyproofness constraint, we require truthfulness-in-expectation, i.e. a guarantee that no agent can reduce her expected cost from misreporting. 1.1 Our results the street). We prove that the approximation ratio of Mechanism M2 for the social cost is max{n − 1, 1 + 2b/c}, where n is the number of agents and conjecture that no deterministic strate- gyproof mechanism can achieve an o(n) approximation ratio in this case. For the maximum cost, the ratio of Mechanism M2 is max{1 + 2b/c, 3} which means that the mechanism is actually the best in the natural class of anonymous and position invariant mechanisms. For any deterministic strategyproof mechanism, we prove a lower bound of 2 on the approximation ratio, proving that at least for the natural case of b = c, Mechanism M2 is also not far from optimal. Finally, we prove an impossibility result; there is no (strongly) group strategyproof, anonymous and position invariant mechanism for the problem. This is in constrast with the single-peaked preference setting, where there is a large class of (strongly) group strategyproof mechanisms that satisfy those properties. Our results are summarized in Table 1. 1.1 Our results Our main contribution is a truthful-in-expectation mechanism (M1) that achieves an approx- imation ratio of 1+b/c for the social cost and max{1+b/c, 2} for the maximum cost, where b is the distance between an agent’s location and her peak and c is her minimum cost. We also prove that no truthful-in-expectation mechanism can do better than a 3/2 approximation for the maximum cost proving that at least for the natural special case where b = c, Mecha- nism M1 is not far from the best possible. For deterministic mechanisms, we prove that no mechanism in a wide natural class of strategyproof mechanisms can achieve an approxima- tion ratio better than 1 + b/c for the social cost and 1 + 2b/c for the maximum cost and hence cannot outperform mechanism M1. To prove this, we ﬁrst characterize the class of strategyproof, anonymous and position invariant mechanisms for two agents, showing that it consists only of a single Mechanism (M2). Intuitively, anonymity requires that all agents are handled equally (irrespectively of their names) by the mechanism while position invariance essentially requires that if we shift an instance by some amount, the location of the facility should be shifted by the same amount as well. This is a quite natural condition and can be 123 123 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1212 Table 1 Summary of our results Double-peaked Single-peaked Upper Lower Upper Lower Social cost Deterministic n −1 1 + b c 1 1 Randomized 1 + b c – 1 1 Maximum cost Deterministic∗ 1 + 2b c 1 + 2b c 2 2 Randomized 1 + b c 3/2 3/2 3/2 The lower bounds for deterministic mechanisms hold for anonymous and position invariant strategyproof h i For (∗), an additional lower bound of 2 holds under no conditions Fields indicated by (−) are not proven yet. For the social cost, the deterministic upper bound is actually max{n −1, 1 + 2b/c} For the maximum cost, the approximation ratios are actually max{1+2b/c, 3} and max{1+b/c, 2} respectively The results for single-peaked preferences are also noted for comparison interpreted as a guarantee that the facility will be located relatively to the reports of the agents and independently of the underlying network (e.g. the street). interpreted as a guarantee that the facility will be located relatively to the reports of the agents and independently of the underlying network (e.g. 1.2 Related work on facility location The strategic version of the facility location problem in computer science was ﬁrst studied by Proccacia and Tennenholtz [30] in a seminal paper, where the authors coined the term approximate mechanism design without money to study problems where, in the absence of monetary terms, strategyproof mechanisms can only achieve the desired objectives within some approximation factor. The main goal of [30] is the location of a single facility on the real line when agents have single-peaked preferences, and the corresponding bounds shown in Table 1 are from that paper. A series of papers have studied generalizations of the problem to more general metric spaces [1,11,32], multiple facilities [14,23,26,27] or even enhancing strategyproof mechanisms with additional capabilities [21,22]. Most of the related work actually considers the same objectives that we do here, namely the social cost or the maximum cost, with the notable exceptions of the least-squares objective [18], the L p norm of costs [17] or the minimax envy [5]. In a recent paper, Procaccia et al. [29] use the facility location problem to explore the trade-offs between the approximation guarantees and the variance of truthful-in-expectation mechanisms. 123 1213 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 In the literature of artiﬁcial intelligence and multi-agent systems, in recent years, several variations of the basic model have emerged, capturing different situations that may arise in practice. A series of papers [7,8,16] study the obnoxious facility location problem over an interval, where agents declare their least preferred point and their utility increases linearly in both directions away from that point. Preference relations that give rise to such utility struc- tures are referred to as single-dipped or single-caved preferences [25]. Seraﬁno and Ventre [33,34] introduced and studied the setting with heterogeneous facilities, in which agents’ locations are known and they declare their interests in different facilities. Building on the idea of heterogeneous facilities, a recent literature [16,39] considers a single facility location setting where the agents report their most preferred positions, along with a binary variable, indicating whether the facility is desirable or obnoxious. Preference proﬁles in such a setting can be seen as being a combination of single-peaked and single-dipped preference orderings. In a different direction, Todo et al. [37] and Sonoda et al. [35] study the capabilities of false- name proof mechanisms, which are strategyproof mechanisms that are also impervious to agents assuming false identities. 1.2 Related work on facility location In light of these recent developments, the current paper (the conference version of which preceded many of the aforementioned papers) can be seen as a different generalization of the basic facility location model, capturing different real-life scenarios and building on the vastly growing literature on such extensions. 2 Preliminaries Let N = {1, 2, . . . , n} be a set of agents. We consider the case where agents are located on a line, i.e. each agent i ∈N has a location xi ∈R. We will occasionally use xi to refer to both the position of agent i and the agent herself. We will call the collection x = ⟨x1, . . . , xn⟩a location proﬁle or an instance. A deterministic mechanism is a function f : Rn →R that maps a given location proﬁle to a point in R, the location of the facility. We assume that agents have double-peaked preferences, symmetric with respect to the origin. We discuss how our results extend to non- symmetric agents in Sect. 6. Given any instance x and a location y ∈R, the cost of agent i is cost(y, xi) = c + \|xi −b −y\| if y ≤xi c + \|xi + b −y\| if y > xi where c and b are positive constants. We will say that y admits a cost of cost(y, xi) for agent i on instance x. For a mechanism that outputs f (x) on instance x, the cost of agent i is cost( f (x), xi). Intuitively, each agent has two most favorable locations, i.e. xi −b and xi +b, which we refer to as the peaks of agent i. Note that these peaks are actually the troughs of the curve of the cost function, but much like most related work, we refer to them as peaks. The parameter c > 0 is the minimum cost incurred to an agent when the facility is built on one of her peaks.1 Note that the special case, where b = c corresponds to the natural setting where the incurred minimum cost of an agent is interpreted as the distance she needs to cover to actually reach the facility. This case is particularly appealing, since the bounds we obtain are clean numbers, independent of b and c. The bounds for the natural case can be obtained directly by letting b = c in all of our results. A randomized mechanism is a function f : Rn →Δ(R), where Δ(R) is the set of probability distributions over R. It maps a given location proﬁle to probabilistically selected locations of the facility. 1 It is not hard to see by our results that if we let an agent’s cost be zero on her peaks, then in very general settings, no determnistic strategyproof mechanism can guarantee a ﬁnite approximation ratio. 1.3 Related work on double-peaked preferences Egan provides additional evidence for the occurrence of double-peaked 123 1214 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 preferences supported by experimental results based on surveys on the U.S. population, for many different problems (education, health care, illegal immigration treatment, foreign oil treatment etc). More examples of double-peaked preferences in real-life scenarios are presented in [31]. The related work demonstrates that although they might not be as popular as their single- peaked counterpart, double-peaked preferences do have applications in settings more general than the street example described earlier. On the other hand, the primary focus of this paper is to study double-peaked preferences on facility location settings and therefore the modeling assumptions follow the ones of the facility location literature. In the literature of multi-agent systems, Yang and Guo [38] consider κ-peaked prefer- ences (a straightforward generalization of double-peaked preferences) and the problem of controlling elections for such preference proﬁles for several well-known voting rules, proving several hardness and parametrized complexity results. 1.3 Related work on double-peaked preferences Single-peaked preferences were introduced in [4] as a way to avoid Condorcet cycles in majority elections. Moulin [28] characterized the class of strategyproof mechanisms in this setting, proving that median voter schemes are essentially the only strategyproof mechanisms for agents with single-peaked preferences. Double-peaked preferences have been mentioned in social choice literature, to describe settings where preferences are not single-peaked, voting cycles do exist and majority elections are not possible. For example, Cooter ([10], pages 39– 42) argues that when the decisions involve multi-dimensional choices, preferences are more likely to be double-peaked rather than single-peaked and the intransitivity of the preferences disallows for majority winners. As a motivating example, he uses the choice of the level of expenditure in public schools in the U.S. and the example of “yuppies”, i.e. young urban professionals that would prefer either a high expenditure level or a low expenditure level rather than a moderate level, as in the ﬁrst case they could send their children to public school whereas in the second case they would send them to a private school without being burdened by large taxes for the support of public education. A simpler example of the same principle can be the choice of temperature in a room; both 15◦with a lightweight jacket and 25◦with just a regular shirt might be preferable to a moderate choice of 20◦. More broadly, in social choice settings similar to the example of [10], double-peaked preferences can be used to model situations where e.g. a left-wing party might prefer a more conservative but quite effective policy to a more liberal but ineffective one on a left-to-right political axis. In fact, Egan [13] provides a detailed discussion on double-peaked preferences in political decisions. He uses a 1964–1970 survey about which course of action the United States should take with regard to the Vietnam war as an example where the status quo (keep U.S. troops in Vietnam but try to terminate the war) was ranked last by a considerable fraction of the population when compared to a left-wing policy (pull out entirely) or a right- wing policy (take a stronger stand). This demonstrates that in a scenario where the standard approach would be to assume that preferences are single-peaked, preferences can instead be double-peaked. 2 Preliminaries Given an instance x and a location y ∈R, the social cost and the maximum cost of y are deﬁned respectively as: SCy(x) = n i=1 cost(y, xi), MCy(x) = max i∈N cost(y, xi). We will say that y admits a social cost of SCy(x) or a maximum cost of MCy(x). We will call y ∈R an optimal location (for the social cost), if y ∈arg miny SCy(x). The deﬁnition for the maximum cost is analogous. Let SCopt(x) and MCopt(x) denote the social cost and the maximum cost of an optimal location respectively, on instance x. For a mechanism f that outputs f (x) on instance x, we will call SCf(x)(x) the social cost of the mechanism and we will denote it by SCf(x); and analogously for the maximum cost. We are interested in strategyproof mechanisms that perform well with respect to the goal of minimizing either the social cost or the maximum cost. We measure the perfor- mance of the mechanism by comparing the social/maximum cost it achieves with the optimal social/maximum cost, on any instance x. The approximation ratio of mechanism f , with respect to the social cost, is given by r = sup x SCf(x) SCopt(x). r = sup x SCf(x) SCopt(x). r = sup x SCf(x) SCopt(x). The approximation ratio of mechanism f , with respect to maximum cost, is deﬁned similarly. F d i d h i th d ﬁiti i il d th i ti ti i The approximation ratio of mechanism f , with respect to maximum cost, is deﬁned similarly. For randomized mechanisms, the deﬁnitions are similar and the approximation ratio is calculated with respect to the expected social or maximum cost, i.e. the expected sum of For randomized mechanisms, the deﬁnitions are similar and the approximation ratio is calculated with respect to the expected social or maximum cost, i.e. the expected sum of costs of all agents and expected maximum cost of any agent, respectively. Finally we consider some properties which are quite natural and are satisﬁed by many mechanisms (including the optimal mechanism). A mechanism f is anonymous, if for every location proﬁle x and every permutation π of the agents, f (x1, . . . , xn) = f (xπ(1), . . . , xπ(n)). 2 Preliminaries The expected cost of agent i is Ey∼D [cost(y, xi)], where D is the probability distribution of the mechanism outputs. We will call a deterministic mechanism f strategyproof if no agent would beneﬁt by misreporting her location, regardless of the locations of the other agents. This means that for 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1215 every x ∈Rn, every i ∈N and every x′ i ∈R, cost( f (x), xi) ≤cost( f (x′ i, x−i), xi), where x−i = ⟨x1, . . . , xi−1, xi+1, . . . , xn⟩. A mechanism is truthful-in-expectation if it guarantees that every agent always minimizes her expected cost by reporting her location truthfully. Throughout the paper we will use the term strategyproofness when refering to deterministic mechanisms and the term truthfulness when refering to randomized mechanisms. every x ∈Rn, every i ∈N and every x′ i ∈R, cost( f (x), xi) ≤cost( f (x′ i, x−i), xi), where x−i = ⟨x1, . . . , xi−1, xi+1, . . . , xn⟩. A mechanism is truthful-in-expectation if it guarantees that every agent always minimizes her expected cost by reporting her location truthfully. Throughout the paper we will use the term strategyproofness when refering to deterministic mechanisms and the term truthfulness when refering to randomized mechanisms. A mechanism is (strongly) group strategyproof if there is no coalition of agents, who by jointly misreporting their locations, affect the outcome in a way such that the cost of none of them increases and the cost of at least one of them strictly decreases. In other words, there is no S ⊆N such that for some misreports xS of agents in S and some reports x−S of agents in N\S, cost( f (x′ S, x−S), xi) ≤cost( f (x), xi) for all i ∈S, and cost( f (x′ S, x−S), x j) < cost( f (x), x j) for at least one j ∈S. We note here that the deﬁnition above is often referred in the literature as strong group strategyproofness, to distinguish it from weak group strategyproofness, where for a group deviation to be possible, it should be the case that the cost of all agents of the deviating coalition strictly decreases. Throughout the paper, when referring to group strategyproofness, we will assume that the notion follows the strong deﬁnition. 2 Preliminaries We say that a mechanism f is onto, if for every point y ∈R on the line, there exists a location proﬁle x such that f (x) = y. Without loss of generality, for anonymous mechanisms, we can assume x1 ≤· · · ≤xn. A property that requires special mention is that of position invariance, which is a very natural property as discussed in the introduction. This property was independently deﬁned by [17] where it was referred to as shift invariance. One can view position invariance as an 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1216 analogue to neutrality in problems like the one studied here, where there is a continuum of outcomes instead of a ﬁnite set. Deﬁnition 1 A mechanism f satisﬁes position invariance, if for all location proﬁles x = ⟨x1, . . . , xn⟩and t ∈R, it holds f (x1 + t, x2 + t, . . . , xn + t) = f (x) + t. In this case, we will call such a mechanism position invariant. We will refer to instances x and ⟨x1 + t, x2 + t, . . . , xn + t⟩as position equivalent. Note that position invariance implies the onto condition. Indeed, for any location proﬁle x, with f (x) = y, we have f (x1 + t, x2 + t, . . . , xn + t) = y′ = y + t for any t ∈R, so every point y′ ∈R is a potential output of the mechanism. 3 A truthful-in-expectation mechanism This time, it might be that \|(xi +b)−(x′ m −b)\| < \|(xi +b)−(xm −b)\| but since (x′ m −b) −(xm −b) = (x′ m + b) −(xm + b), it will also hold that \|(xi + b) −(x′ m + b)\| > \|(xi + b) −(xm + b)\| and also \|(xi + b) −(xm −b)\| −\|(xi + b) −(x′ m −b)\| = \|(xi + b) − (x′ m + b)\| −\|(xi + b) −(xm + b)\|. Hence, the expected cost of agent i after misreporting is at least as much as it was before. Again, the cost of agent i is calculated with respect to xi + b for both outcomes of the mechanism. This time, it might be that \|(xi +b)−(x′ m −b)\| < \|(xi +b)−(xm −b)\| but since (x′ m −b) −(xm −b) = (x′ m + b) −(xm + b), it will also hold that \|(xi + b) −(x′ m + b)\| > \|(xi + b) −(xm + b)\| and also \|(xi + b) −(xm −b)\| −\|(xi + b) −(x′ m −b)\| = \|(xi + b) − ( ′ b) ( b) ( b) h d f f i i i Again, the cost of agent i is calculated with respect to xi + b for both outcomes of the mechanism. This time, it might be that \|(xi +b)−(x′ m −b)\| < \|(xi +b)−(xm −b)\| but since ( ′ b) ( b) ( ′ + b) ( + b) it ill l h ld th t \|( + b) ( ′ + b)\| mechanism. This time, it might be that \|(xi +b) (xm b)\| < \|(xi +b) (xm b)\| but since (x′ m −b) −(xm −b) = (x′ m + b) −(xm + b), it will also hold that \|(xi + b) −(x′ m + b)\| > \|(xi + b) −(xm + b)\| and also \|(xi + b) −(xm −b)\| −\|(xi + b) −(x′ m −b)\| = \|(xi + b) − (x′ m + b)\| −\|(xi + b) −(xm + b)\|. Hence, the expected cost of agent i after misreporting is at least as much as it was before. Case 3 xm < xi + b ≤xm + b (symmetrically xm −b ≤xi −b < xm). 3 A truthful-in-expectation mechanism We start the exposition of our results with our main contribution, a truthful-in-expectation mechanism that achieves an approximation ratio of 1 + b/c for the social cost and max{1 + b/c, 2} for the maximum cost. Mechanism M1 Given any instance x = ⟨x1, . . . , xn⟩, ﬁnd the median agent xm = median(x1, . . . , xn), breaking ties in favor of the agent with the smallest index. Output f (x) = xm −b with probability 1 2 and f (x) = xm + b with probability 1 2. Theorem 1 Mechanism M1 is truthful-in-expectation. Theorem 1 Mechanism M1 is truthful-in-expectation. Proof First, note that the median agent does not have an incentive to deviate, since her expected cost is already minimum, neither does any agent i for which xi = xm. Hence, for the deviating agent i it must be either xi < xm or xi > xm. We consider three cases when xi < xm. The proof for the case xi > xm is symmetric. Observe that for agent i to be able to move the position of the facility, she has to report x′ i ≥xm and change the identity of the median agent. Let x′ m be the median agent in the new instance ⟨x′ i, x−i⟩, after agent i’s deviation. If x′ m = xm, then obviously agent xi does not gain from deviating, so we will assume that x′ m > xm. Case 1 xi + b ≤xm −b (symmetrically xi −b ≥xm + b). In this case, the cost of agent i is calculated with respect to xi + b for both possible outcomes of the mechanism. Since x′ m −b > xm −b and x′ m + b > xm + b, it holds that \|(xi +b)−(x′ m −b)\| > \|(xi +b)−(xm −b)\| and \|(xi +b)−(x′ m +b\|) > \|(xi +b)−(xm +b)\| and agent i can not gain from misreporting. Case 2 xm −b < xi + b ≤xm (symmetrically xm ≤xi −b < xm + b). Case 2 xm −b < xi + b ≤xm (symmetrically xm ≤xi −b < xm + b) Again, the cost of agent i is calculated with respect to xi + b for both outcomes of the mechanism. 3.1 Social cost Next, we will calculate the approximation ratio of the mechanism for the social cost. In order to do that, we will need the following lemma. Lemma 1 Let x = ⟨x1, . . . , xm, . . . , xn⟩, where xm = median (x1, . . . , xn), breaking ties in favor of the smallest index. There exists an optimal location for the social cost in [xm − b, xm + b]. Proof Assume by contradiction that there exists a point y < xm −b or y > xm + b with a strictly smaller social cost than all the points in [xm −b, xm + b]. Assume ﬁrst that y < xm −b. Since xm is the median agent, it holds that for at least ⌈n/2⌉agents, xi −b ≥xm −b, that is xm −b admits a smaller cost for at least ⌈n/2⌉ agents when compared to y. Let X1 be the set of those agents. On the other hand, for each agent xi < xm, xm −b may admit a smaller or larger cost than y, depending on her position with respect to y. In the worst case, the cost is larger for every one of those agents, which happens when xi + b ≤y for every agent with xi < xm. Let X2 be the set of those agents. Now observe that for any two agents xα ∈X1 and xβ ∈X2, it holds that cost(xα, y) −cost(xa, xm −b) = cost(xβ, xm −b) −cost(xβ, y). Since \|X1\| ≥\|X2\|, it holds that that SCxm−b(x) ≤SCy(x). Since it holds that SCxm−b(x) > SCy(x), xm −b is an optimal location and we get a contradiction. Now assume y > xm + b. If the number of agents is odd, then we can use an exactly symmetric argument to prove that SCxm+b ≤SCy. If the number of agents is even, the argument can still be used, since our tie-breaking rule selects agent xn/2 as the median. Speciﬁcally, xm +b admits a smaller cost for exactly n/2 of the agents (including agent xn/2) and in the worst case, y admits a smaller cost for n/2 agents as well. If X1 and X2 are the sets of those agents respectively, then again it holds that cost(xα, y) −cost(xα, xm + b) = cost(xβ, xm + b) −cost(xβ, y) for xα ∈X1 and xβ ∈X2 and we get a contradiction as before. 3 A truthful-in-expectation mechanism The cost of agent i before misreporting is calculated with respect to xi −b when the outcome is xm −b and with respect to xi + b when the outcome is xm + b. For any misreport 123 1217 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 x′ i < xi + b, this is still the case (for x′ m −b and x′ m + b respectively) and since (x′ m − b) −(xm −b) = (x′ m + b) −(xm + b), her expected cost is not smaller than before. For any misreport x′ i > xi +b, her cost is calculated with respect to xi +b for both possible outcomes of the mechanism and for the same reason as in Case 2, her expected cost is at least as much as it was before misreporting. ⊓⊔ x′ i < xi + b, this is still the case (for x′ m −b and x′ m + b respectively) and since (x′ m − b) −(xm −b) = (x′ m + b) −(xm + b), her expected cost is not smaller than before. For any misreport x′ i > xi +b, her cost is calculated with respect to xi +b for both possible outcomes of the mechanism and for the same reason as in Case 2, her expected cost is at least as much as it was before misreporting. ⊓⊔ 3.1 Social cost ⊓⊔ We now proceed to proving the approximation ratio of Mechanism M1. Theorem 2 Mechanism M1 has an approximation ratio of 1 + b c for the social cost. Proof Consider an arbitrary instance x = ⟨x1, . . . , xn⟩and let xm be the median agent. By Lemma 1, there exists an optimal location y ∈[xm −b, xm + b]. Let δ = y −(xm −b). For every agent i, it holds that cost(xi, xm −b) ≤cost(xi, y) + δ. To see this, ﬁrst observe that \|(xi −b)−(xm −b)\| ≤\|(xi −b)−y\|+δ and that \|(xi +b)−(xm −b)\| ≤\|(xi +b)−y\|+δ. If the cost of an agent admitted by y and xm −b is calculated with respect to the same peak, then min(\|(xi −b)−(xm−b)\|, \|(xi +b)−(xm−b)\|) ≤min(\|(xi −b)−y\|, \|(xi +b)−y\|)+δ and the inequalityholds.Ifthecostiscalculatedwithrespecttodifferentpeaksfor y and xm−b,itmust be that cost(xi, xm −b) = c+\|(xi −b)−(xm −b)\| and cost(xi, y) = c+\|xi +b−y\|, because xm−b < y.Since\|(xi−b)−(xm−b)\| ≤\|(xi+b)−(xm−b)\| ≤\|(xi+b)−y\|+δ,theinequality holds. Similarily, we can prove that cost(xi, xm +b) ≤cost(xi, y)+(2b −δ) for every agent 123 1218 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 i. Hence, we can upper bound the cost of Mechanism M1 by 1 2 n i=1 cost(xi, xm −b)+ 1 2 n i=1 cost(xi, xm + b) ≤1 2 n i=1 (cost(xi, y) + δ) + 1 2 n i=1 (cost(xi, y) + 2b −δ) = SCy(x) + nb = SCopt(x) + nb. The approximation ratio then becomes 1 + nb SCopt(x), which is at most 1 + b c , since SCopt(x) is at least nc. c p For the lower bound, consider the location proﬁle x = ⟨x1, . . . , xn⟩with x1 = · · · = xk−1 = xk −b = xk+1 −2b = · · · = xn −2b. Note that the argument works both when n = 2k and when n = 2k + 1 because Mechanism M1 selects agent xk as the median agent in each case. The optimal location is x1 + b whereas Mechanism M1 equiprobably outputs fM1(x) = xk −b or fM1(x) = xk + b. The cost of the optimal location is SCopt(x) = nc + b whereas the cost of Mechanism M1 is SCM1(x) = nc + (1/2)(n −1)b + (1/2)(n −1)b = nc + (n −1)b. The approximation ratio then becomes nc+(n−1)b nc+b = 1 b c · n−2 n+(b/c). As the number of agents grows to inﬁnity, the approximation ratio of the mechanism on this instance approaches 1 + b/c. This completes the proof. ⊓⊔ 3.2 Maximum cost We also consider the maximum cost and prove the approximation ratio of Mechanism M1 as well as a lower bound on the approximation ratio of any truthful-in-expectation mechanism. The results are summarized in Table 1. Theorem 3 Mechanism M1 has an approximation ratio of max{1+b/c, 2} for the maximum cost. Proof Let x = ⟨x1, . . . , xn⟩be an arbitrary instance and let xm be the median agent. We will consider two cases, based on the location of fopt(x) with respect to xm −b (or symmetrically xm + b). Case 1 fopt(x) < xm −b (or fopt(x) > xm + b). Case 1 fopt(x) < xm −b (or fopt(x) > xm + b). Let δ = (xm −b) −fopt(x). For the same reason as in the proof of Theorem 2, for every agent i, it holds that cost(xi, xm −b) ≤cost(xi, fopt(x))+δ and also that cost(xi, xm +b) ≤ cost(xi, fopt(x)) + (2b + δ). p The maximum cost of Mechanism M1 is 1 1 p The maximum cost of Mechanism M1 is MCM1(x) = 1 2 max i∈N cost (xi, xm −b) + 1 2 max i∈N cost (xi, xm + b) ≤1 2 max i∈N cost xi, fopt(x) + δ + 1 2 max i∈N cost xi, fopt(x) + (2b + δ) = MCopt(x) + b + δ ≤2MCopt(x) + b −c The maximum cost of Mechanism M1 is MCM1(x) = 1 2 max i∈N cost (xi, xm −b) + 1 2 max i∈N cost (xi, xm + b) ≤1 2 max i∈N cost xi, fopt(x) + δ + 1 2 max i∈N cost xi, fopt(x) + (2b + δ) = MCopt(x) + b + δ ≤2MCopt(x) + b −c since MCopt(x) ≥c + δ. The approximation is at most 2 + b−c MCopt(x), which is at most 1 + b c if b ≥c (since MCopt(x) ≥c) and at most 2 if b < c (since MCopt(x) > 0). Case 2 xm −b ≤fopt(x) ≤xm + b. Case 2 xm −b ≤fopt(x) ≤xm + b. Now, let δ = fopt(x)−(xm−b). Again, it holds that cost(xi, xm−b) ≤cost(xi, fopt(x))+δ and also that cost(xi, xm + b) ≤cost(xi, fopt(x)) + (2b −δ). 3.2 Maximum cost p The maximum cost of Mechanism M1 is MCM1(x) = 1 2 max i∈N cost (xi, xm −b) + 1 2 max i∈N cost (xi, xm + b) MCM1(x) = 1 2 max i∈N cost (xi, xm −b) + 1 2 max i∈N cost (xi, xm + b) ≤1 2 max i∈N cost xi, fopt(x) + δ + 1 2 max i∈N cost xi, fopt(x) + (2b −δ) = MCopt(x) + b ≤1 2 max i∈N cost xi, fopt(x) + δ + 1 2 max i∈N cost xi, fopt(x) + (2b −δ) 123 123 1219 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 and the approximation ratio is at most 1 + b c (since MCopt(x) ≥c). and the approximation ratio is at most 1 + b c (since MCopt(x) ≥c). and the approximation ratio is at most 1 + b c (since MCopt(x) ≥c). and the approximation ratio is at most 1 + b c (since MCopt(x) ≥c). c p For the matching lower bound, consider an instance x = ⟨x1, . . . , xn⟩on which x1 + b < x2 −b and xi = x2 for all i /∈{1, 2}. It is fopt(x) = x1+x2 2 , i.e. the middle of the interval between x1 and x2, whereas Mechanism M1 selects equiprobably among x2−b and x2+b. Let d = fopt(x)−(x1+b).Then MCopt(x) = c+d,whereas MCM1(x) = c+ 1 22d+ 1 2(2d+2b) = b + c + 2d. The approximation ratio is 1 + b+d c+d which is 1 + b c as d goes to 0 and 2 as d goes to inﬁnity. ⊓⊔ ⊓⊔ Next, we provide a lower bound on the approximation ratio of any truthful-in-expectation mechanism. Theorem 4 For any values of c and b, no truthful-in-expectation mechanism can achieve an approximation lower than 3 2 for the maximum cost. First, we state a couple of lemmas which are in essence very similar to those used in the proof of the single-peaked preferences case in [30]. Let x = ⟨x1, x2⟩be an instance such that x1 + b < x2 −b and let λ = (x2 −b) −(x1 + b). Let f be a truthful-in-expectation mechanism and let D be the distribution that y = f (x) follows on instance x. 4 Deterministic mechanisms We now turn our attention to deterministic mechanisms. We will start by stating and proving the following lemma, which will be very useful throughout the paper. Variations of the instances used here will appear in several of our proofs. When n = 2, we deﬁne the following family of instances, called primary instances. Primary instance We will say that an instance is a primary instance, if it holds that x = ⟨x1, x2⟩with x1 +2b+ϵ = x2 −b, where ϵ is a positive real number. In the following we will ﬁx such an ϵ > 0 (e.g. ϵ = b/2) and refer to the resulting instance as the primary instance. Lemma 4 On the primary instance, there is no anonymous, position invariant and strate- gyproof mechanism such that f (x) ∈[x1 + b, x2 −b]. Proof For contradiction, suppose there exists an anonymous, position invariant strategyproof mechanism M that ouputs f (x) ∈[x1 + b, x2 −b]. Let’s denote δ1 = f (x) −(x1 + b), δ2 = (x2 −b)−f (x). Throughout the proof we start with the primary instance and construct some other instances to prove the lemma. There are 2 cases to be considered. Case 1 0 < δ1 ≤ 1 2(b + ϵ), i.e. x1 + b < f (x) ≤ x1+x2 2 (symmetrically, for the case 0 < δ2 ≤1 2(b + ϵ)). See Fig. 1. – Instance I : xI = ⟨xI 1, xI 2⟩, where xI 1 = x1 + δ1, xI 2 = x2. – Instance II : xII = ⟨xII 1 , xII 2 ⟩, where xII 1 = x1 + δ1, xII 2 = x2 + δ1. First, we know that on instance I, strategyproofness requires that f (xI) = xI 1−b or xI 1+b, otherwise agent xI 1 could misreport x1 and move the facility to position xI 1 + b, minimizing her cost. Second, since instance II and the primary instance are position equivalent (II is a shifted version of the primary instance to the right by δ1), by position invariance we know f (xII) = xII 1 + b + δ1 = xI 1 + b + δ1. Now let’s consider agent xI 2 in instance I. 3.2 Maximum cost Lemma 2 On instance x, at least one of Ey∼D [cost(x1, y)] ≥c+ λ 2 and Ey∼D [cost(x2, y)] ≥c + λ 2 holds. Proof Obviously, cost(x1, y) + cost(x2, y) ≥ 2c + λ for any choice of y, hence Ey∼D 2 i=1 cost(xi, y) = 2 i=1 Ey∼D[cost(xi, y)] ≥2c + λ. Therefore, it must be that Ey∼D[cost(xi, y)] ≥c + λ 2 for at least one of i = 1 or i = 2. ⊓⊔ Lemma 3 Let fopt(x) be the outcome of the optimal mechanism on instance x. If Ey∼D[\|y − fopt(x)\|] = Δ, then the maximum cost of the mechanism on this instance is E[MC f (x)] = c + λ 2 + Δ. Proof Since ∀y, MC f (x) = c + \|y −fopt(x)\| + λ/2, it holds Ey∼D MC f (x) = Ey∼D c + λ 2 + y −fopt(x) = c + λ 2 + Ey∼D y −fopt(x) = c + λ 2 + Δ. ⊓⊔ ⊓⊔ We can now prove the theorem. We can now prove the theorem. e can now prove the theorem. Proof Consideraninstancewithtwoagentsonwhich x1+b < x2−b and(x2−b)−(x1+b) = λ. It holds that fopt(x) = x1+x2 2 . Assume there is a truthful-in-expectation mechanism M on which y = f (x1, x2) follows a distribution D on this instance. According to Lemma 2, at least one of Ey∼D[cost(y, x1)] ≥c + λ/2 and Ey∼D[cost(y, x2)] ≥c + λ/2 holds. W.l.o.g., assume the second inequality is true (if the ﬁrst inequality is true then we can make a symmetric argument with agent x1 deviating). Next, consider the instance x′ = ⟨x′ 1, x′ 2⟩with x′ 1 = x1 and x′ 2 = x2 + λ. Let fopt(x′) = (x′ 1 + x′ 2)/2 = x2 −b. Let D′ be the distribution that y′ follows on instance x′. By strategyproofness, Ey′∼D′ cost(y′, x2) ≥Ey∼D[cost(y, x2)] ≥c + λ/2, since x′ 2 123 123 1220 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 could be a deviation of agent 2 on instance x. This implies Ey′∼D′ \|y′ −(x2 −b)\| ≥λ 2. 3.2 Maximum cost To see this, assume otherwise for contradiction, we would have Ey′∼D′ cost(y′, x2) = Ey′∼D′ c + min y′ −(x2 −b) , y′ −(x2 + b) ≤c + min Ey′∼D′ y′ −(x2 −b) , Ey′∼D′ y′ −(x2 + b) < c + λ 2 . Equivalently, we have Ey′∼D′ \|y′ −fopt(x′)\| ≥λ 2. Hence, by applying Lemma 3, we know that the maximum cost of the mechanism on the second instance is Ey′∼D′ MCM(x′) ≥ c+λ+ λ 2 = c+ 3λ 2 . Since the optimal mechanism locates the facility on fopt(x′), its maximum cost is c + λ. Therefore, the approximation ratio is at least c+3λ/2 c+λ . As λ grows to inﬁnity, the approximation ratio approaches 3 2. To generalize the proof to more than two agents, place every other agent on fopt(x) + b on instance x. Since λ is large enough, the maximum cost is still calculated with respect to x2 and all the arguments still hold. ⊓⊔ Case 2 δ1 = 0, i.e. f (x) = x1 + b (symmetrically, for the case δ2 = 0). See Fig. 2. 4 Deterministic mechanisms If agent xI 2 misreports xII 2 , she can move the facility from xI 1 −b or xI 1 + b to xI 1 + b + δ1 ≤xI 2 −b; in either case the facility is closer to her left peak. Therefore xI 2 can manipulate the mechanism, which violates strategyproofness. Hence f (x) /∈(x1 + b, x1+x2 2 ]. Case 2 δ1 = 0, i.e. f (x) = x1 + b (symmetrically, for the case δ2 = 0). See Fig. 2. Case 2 δ1 = 0, i.e. f (x) = x1 + b (symmetrically, for the case δ2 = 0). See Fig. 2. 123 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1221 Auton Agent Multi Agent Syst (2017) 31:1209 1235 1221 Fig. 1 Case 1 of Lemma 4. On instance I, agent xI 2 can report xII 2 and decrease her cost. Instances II and the Primary instance are position equivalent Fig. 2 Case 2(a) and 2(b) of Lemma 4. If f (xIII 1 , xIII 2 ) = xIII 1 −b on instance III, agent xIII 2 can misreport xIVa 2 and decrease her cost, since instances IVa and the Primary instance are position equivalent. If f (xIII 1 , xIII 2 ) = xIII 1 + b on instance III, then on the Primary instance, agent x2 can misreport xIVb 2 and decrease her cost. By anonymity, instances III and IVb are position equivalent Fig. 1 Case 1 of Lemma 4. On instance I, agent xI 2 can report xII 2 and decrease her cost. Instances II and the Primary instance are position equivalent Fig. 1 Case 1 of Lemma 4. On instance I, agent xI 2 can report xII 2 and decrease her cost. Instances II and the Primary instance are position equivalent Fig. 1 Case 1 of Lemma 4. On instance I, agent xI 2 can report xII 2 and decrease her cost. Instances II and the Primary instance are position equivalent Primary instance are position equivalent Fig. 2 Case 2(a) and 2(b) of Lemma 4. If f (xIII 1 , xIII 2 ) = xIII 1 −b on instance III, agent xIII 2 can misreport xIVa 2 and decrease her cost, since instances IVa and the Primary instance are position equivalent. 4 Deterministic mechanisms Obviously, instance IVb is position equivalent to instance III, so it holds that f (xIVb 1 , xIVb 2 ) = xIVb 2 +b as implied by anonymity and position invariance. Note that on instance III, the cost of agent xIII 2 is b +c −ϵ. If agent xIII 2 misreports xIVb 2 , then her cost becomes c + 2ϵ, which is smaller than when reporting truthfully (since ϵ is arbitrarily small). So f (xIII) ̸= xIII 1 + b if Mechanism M is strategyproof. 1 (b) f (xIII) = xIII 1 + b. See right hand side of Fig. 2, where we have instance IVb: xIVb 1 = xIII 1 , xIVb 2 = xIII 1 −(b + ϵ). Obviously, instance IVb is position equivalent to instance III, so it holds that f (xIVb 1 , xIVb 2 ) = xIVb 2 +b as implied by anonymity and position invariance. Note that on instance III, the cost of agent xIII 2 is b +c −ϵ. If agent xIII 2 misreports xIVb 2 , then her cost becomes c + 2ϵ, which is smaller than when reporting truthfully (since ϵ is arbitrarily small). So f (xIII) ̸= xIII 1 + b if Mechanism M is strategyproof. e, in order for mechanism M to be strategyproof, it must be f (x) ̸= x1 + b. Hence, in order for mechanism M to be strategyproof, it must be f (x) ̸= x1 + b. In all, there is no anonymous, position invariant and strategyproof mechanism such that f (x) ∈[x1 + b, x2 −b]. ⊓⊔ In all, there is no anonymous, position invariant and strategyproof mechanism such that f (x) ∈[x1 + b, x2 −b]. ⊓⊔ It is well known [28] that when agents have single-peaked preferences, outputting the location of the kth agent (kth order statistic), results in a strategyproof mechanism. This is not the case however, for double-peaked preferences and any choice of k. Lemma 5 Given any instance x = ⟨x1, . . . , xn⟩, any mechanism that outputs f (x) = xi −b, for i = 2, . . . , n or any mechanism that outputs f (x) = xi + b, for i = 1, . . . , n −1, is not strategyproof. Proof We prove the case when f (x) = x1 +b. The arguments for the other cases are similar. 4 Deterministic mechanisms If f (xIII 1 , xIII 2 ) = xIII 1 + b on instance III, then on the Primary instance, agent x2 can misreport xIVb 2 and decrease her cost. By anonymity, instances III and IVb are position equivalent Fig. 2 Case 2(a) and 2(b) of Lemma 4. If f (xIII 1 , xIII 2 ) = xIII 1 −b on instance III, agent xIII 2 can misreport xIVa 2 and decrease her cost, since instances IVa and the Primary instance are position equivalent. If f (xIII 1 , xIII 2 ) = xIII 1 + b on instance III, then on the Primary instance, agent x2 can misreport xIVb 2 and decrease her cost. By anonymity, instances III and IVb are position equivalent – Instance III : xIII = ⟨xIII 1 , xIII 2 ⟩, where xIII 1 = x1 + 2b, xIII 2 = x2. First, we know that on instance III, strategyproofness requires either that f (xIII) = xIII 1 −b or f (xIII) = xIII 1 +b, otherwise agent xIII 1 could misreport x1 and move the facility to position xIII 1 −b, minimizing her cost. Now, there are two subcases. First, we know that on instance III, strategyproofness requires either that f (xIII) = xIII 1 −b or f (xIII) = xIII 1 +b, otherwise agent xIII 1 could misreport x1 and move the facility to position xIII 1 −b, minimizing her cost. Now, there are two subcases. 123 1222 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 (a) f (xIII) = xIII 1 −b. See left hand side of Fig. 2, where we have instance IVa: xIVa 1 = x1 + 2b, xIVa 2 = x2 + 2b. Obviously, instance IVa is position equivalent to the primary instance, so f (xIVa 1 , xIVa 2 ) = xIVa 1 + b. Note that the cost of agent xIII 2 is b + c + ϵ. If agent xIII 2 misreports xIVa 2 , then her cost becomes b + c −ϵ, which is smaller than when reporting truthfully. So f (xIII) ̸= xIII 1 −b if Mechanism M is strategyproof. III III f ̸ 1 gyp (b) f (xIII) = xIII 1 + b. See right hand side of Fig. 2, where we have instance IVb: xIVb 1 = xIII 1 , xIVb 2 = xIII 1 −(b + ϵ). 4 Deterministic mechanisms Consider any instance where x2 = x1 + b and xi −b > x2 + b for i = 3, . . . , n. Since f (x) = f (x1, x2, x3, . . . , xn) = x1+b, the cost of agent 2 is cost( f (x), x2) = b+c. If agent 2 misreports x′ 2 = x1 −b, the outcome will be f (x′) = f (x1, x′ 2, x3, . . . , xn) = x′ 2 +c = x1, and cost( f (x′), x2) = c < b + c = cost( f (x), x2). Agent 2 has an incentive to misreport, therefore the mechanism is not strategyproof. ⊓⊔ This only leaves two potential choices among kth order statistics, either f (x) = x1 −b or f (x) = xn + b. Consider the following mechanism. Mechanism M2 Given any instance x = ⟨x1, . . . , xn⟩, locate the facility always on the left peak of agent 1, i.e. f (x) = x1 −b or always on the right peak of agent n, i.e. f (x) = xn +b. From now on, we will assume that M2 locates the facility on x1 −b on any instance x. The analysis for the other case is similar. Theorem 5 Mechanism M2 is strategyproof. Theorem 5 Mechanism M2 is strategyproof. Proof Obviously, agent 1 has no incentive to misreport, since her cost is already minimum. For any other agent i, i = 2, . . . , n, the cost is cost( f (x), xi) = c+xi −x1. For any misreport x′ i ≥x1, the facility is still located on x1 −b and every agent’s cost is the same as before. For some misreport x′ i < x1, the facility moves to f (x1, . . . , x′ i, . . . , xn) = x′ i −b < x1 −b, i.e. further away from any of agent i’s peaks and hence this choice admits a larger cost for her. ⊓⊔ 123 1223 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 In the following, we prove that for the case of two agents, Mechanism M2 is actually the only strategyproof mechanism that satisﬁes anonymity and position invariance. We start with the following lemma. Lemma 6 For any instance x = ⟨x1, x2⟩, where x1 + b < x2 −b, if an anonymous, position invariant and strategyproof mechanism outputs f (x1, x2) = x1 −b, then it must output f (x′ 1, x′ 2) = x′ 1 −b for any instance x′ = ⟨x′ 1, x′ 2⟩with x′ 1 ≤x′ 2. Proof Let (x2 −b) −(x1 + b) = γ . For any other instance x′ = ⟨x′ 1, x′ 2⟩with x′ 1 ≤x′ 2, we assume x′ 1 = x1 without loss of generality due to position invariance. In particular, we will ﬁrst prove that on any instance x′ with x′ 1 = x1 and x′ 2 > x1, the position of the facility has to be x′ 1 −b and we will establish that by only considering potential deviations of agent 2 on instance x such that x′ 2 ≥x1. If it holds for this set of deviations, then it certainly holds even if agent 2 can deviate anywhere on the real line. Let S be the set of those instances. But then, any instance ˜x /∈S is position equivalent to some instance x ∈S and therefore it has to be that f (˜x) = ˜x1 −b and the lemma will follow. On instance x, the cost of agent 2 is 2b + c + γ , so for any deviation of x2, her cost must be at least 2b + c + γ , as required by strategyproofness. Theorem 5 Mechanism M2 is strategyproof. This implies that if agent 2 misreports x′ 2, then on the resulting instance it must be either f (x1, x′ 2) ∈(−∞, x1 −b] or f (x1, x′ 2) ∈[x2 + 3b + γ, +∞). First, assume f (x1, x′ 2) ∈[x2 + 3b + γ, ∞). Let instance I be xI = ⟨xI 1, xI 2⟩, where xI 1 = x1, xI 2 = f (x1, x′ 2) + b. On instance I it must be either f (xI 1, xI 2) = f (x1, x′ 2) or f (xI 1, xI 2) = f (x1, x′ 2)+2b, otherwise agent 2 can deviate from xI 2 to x′ 2 and move the facility to xI 2 −b, minimizing her cost and violating strategyproofness. However then, on instance I, agent 1 can misreport ¯xI 1 = xI 2 −2b −γ and move the facility to xI 2 −3b −γ , reducing her cost and violating strategyproofness. This follows from the fact that the resulting instance ⟨¯xI 1, xI 2⟩and instance x are position equivalent and that f (x1, x2) = x1 −b on instance x. Hence, it can not be that f (x1, x′ 2) ∈[x2 + 3b + γ, ∞) on instance x′. 2 Second, assume f (x1, x′ 2) ∈(−∞, x1 −b). Then, since x′ 2 > x1, agent 2 can deviate from x′ 2 to x2 and move the facility to x1 −b, i.e. closer to her actual position. Hence, it can’t be that f (x1, x′ 2) ∈(−∞, x1 −b) on instance x′ either. f 2 In conclusion, it must be f (x1, x′ 2) = f (x′ 1, x′ 2) = x′ 1 −b for any instance x′ = (x′ 1, x′ 2) with x′ 1 ≤x′ 2. ⊓⊔ Theorem 6 When n = 2, the only strategyproof mechanism that satisﬁes position invariance and anonymity is Mechanism M2. cost(y, xi), is not strategyproof. Proof Consider an instance x = ⟨x1, x2, x3⟩, such that x2+b < x3−b, x2−b < x1+b < x2 and (x1+b)−(x2−b) = ϵ, where ϵ is an arbitrarily small positive quantity. On this instance, the optimal facility location is x2 + b and the cost of agent x1 is c + 2b −ϵ. Suppose now that agent x1 reports x′ 1 < x2 −2b. Moreover, suppose that when there are two locations y1 and y2, with y1 < y2 that admit the same social cost, the optimal mechanism outputs y1. If the mechanism outputs y2 instead, we can use a symmetric argument on the instance x′ = ⟨x′ 1, x′ 2, x′ 3⟩= ⟨x′ 1, x2, x2 + 2b −ϵ⟩with agent 3 misreporting x3.2 By this tie-breaking rule, on instance x = ⟨x′ 1, x2, x3⟩, the location of the facility is x2 −b and the cost of agent x1 is c + ϵ, i.e. smaller than before. Hence, the optimal mechanism is not strategyproof. To extend this to an arbitrary number of agents, let x j = x2 for every other agent x j. ⊓⊔ ⊓⊔ Unfortunately, when considering the social cost, in the extremal case, the approximation ratio of Mechanism M2 is dependent on the number of agents. The approximation ratio is given by the following theorem. Theorem 8 For n > 3 agents, the approximation ratio of Mechanism M2 for the social cost is max n −1, 1 + 2b c . max n −1, 1 + 2b c . Proof Consider any instance x′ = ⟨x′ 1, . . . , x′ n⟩and let y = fopt(x′). It also holds that fM2(x′) = x′ 1 −b. Denote the social costs of the optimal mechanism and Mechanism M2 on instancex′ by SCopt(x′)and SCM2(x′),respectively.Letx betheinstanceobtainedbyinstance x′ as follows. For every agent i, i ̸= 1, if x′ i + b ≤y, let xi = 2y −x′ i; if x′ i < y < x′ i + b, let xi = x′ i + 2b; if x′ i −b < y < x′ i, let xi = 2y −x′ i + 2b; otherwise let xi = x′ i. Observe that \|xi −b −y\| = min \|x′ i −b −y\|, \|x′ i + b −y\| and hence cost(xi, y) = cost(x′ i, y) for every agent i. 2 In fact, even if the optimal mechanism outputs a distribution over points that all admit the minimum social cost, the argument still works. Theorem 6 When n = 2, the only strategyproof mechanism that satisﬁes position invariance and anonymity is Mechanism M2. In conclus nymous and position inv gyproof, but how well do cost or the maximum co M2 achieves against the h minimizes the social c respect to the social co ⟩, such that x2+b < x3− rbitrarily small positive q the cost of agent x1 is c eover, suppose that when ame social cost, the opt we can use a symmetric a with agent 3 misreporting tion of the facility is x2 e, the optimal mechanism s, let x j = x2 for every cial cost, in the extremal he number of agents. Th ossible for a strategyproof me y instance. In conclusion, no mous and position invariant. proof, but how well does it pe ost or the maximum cost? In 2 achieves against the optima minimizes the social cost, is spect to the social cost, fopt such that x2+b < x3−b, x2 trarily small positive quantity e cost of agent x1 is c + 2b ver, suppose that when there me social cost, the optimal m can use a symmetric argume h agent 3 misreporting x3.2 B n of the facility is x2 −b an he optimal mechanism is no let x j = x2 for every other a al cost, in the extremal case, number of agents. The appr 4.1 Social cost We have seen that Mechanism M2 is strategyproof, but how well does it perform with respect to our goals, namely minimizing the social cost or the maximum cost? In other words, what is the approximation ratio that Mechanism M2 achieves against the optimal choice? First, we observe that the optimal mechanism, which minimizes the social cost, is not strategyproof. Theorem 6 When n = 2, the only strategyproof mechanism that satisﬁes position invariance and anonymity is Mechanism M2. Proof For contradiction, suppose there exists an anonymous, position invariant strategyproof mechanism M which is different from Mechanism M2 and consider the primary instance used in Lemma 4. We ﬁrst argue that it must be that f (x1, x2) ∈[x1 + b, x2 −b]. Assume on the contrary that f (x1, x2) < x1 + b (the argument for f (x1, x2) > x2 −b is symmetric). Then, consider the instance ⟨x′ 1, x′ 2⟩, where x′ 2 = x2 and x′ 1 = f (x1, x2) −b. On this instance, it must be that f (x′ 1, x′ 2) = x′ 1 −b or f (x′ 1, x′ 2) = x′ 1 + b, otherwise agent 1 can deviate from x′ 1 to x1 and move the facility to x′ 1 + b, minimizing her cost. In addition, if f (x′ 1, x′ 2) = x′ 1−b, then according to Lemma 6, the unique strategyproof mechanism that satisﬁes position invariance and anonymity is Mechanism M2, and thus our assumption is violated. So let’s assume f (x′ 1, x′ 2) = x′ 1 + b. Then, on the primary instance, agent 2 could report ˆx2 = x2 + (x1 + b −f (x1, x2)) and by position invariance (since instances ⟨x′ 1, x′ 2⟩and ⟨x1, ˆx2⟩ are position equivalent), it should be f (x1, ˆx2) = x1 + b. This would give agent 2 an 123 1224 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 incentive to misreport, violating strategyproofness. Hence, on the primary instance it must be that f (x1, x2) ∈[x1 + b, x2 −b]. However, according to Lemma 4, it is impossible for a strategyproof mechanism to output f (x1, x2) ∈[x1 + b, x2 −b] on the primary instance. In conclusion, no mechanism other than Mechanism M2 is strategyproof, anonymous and position invariant. ⊓⊔ mpossible for a strategypr ary instance. cost(y, xi), is not strategyproof. Similarily, it holds that (xi −b) −(x1 −b) ≥(x′ i −b) −(x′ 1 −b) and hence (since x1 = x′ 1), cost(xi −b, x1 −b) ≥cost(x′ i −b, x′ 1 −b) for all agents i. 1 i 1 g We will calculate an upper bound on the approximation ratio on instance x′. To do that, we will calculate an upper bound on the value of the ratio SCM2(x)/SCy(x) on instance x, 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1225 Fig. 3 Cases of Theorem 8. The dotted lines indicate the position of agent xi before moving her from the left side to the right side of y. The cost of every agent with respect to y is the same before and after moving her to the right. The ﬁgure depicts only three agents (including x1 and xn) Fig. 3 Cases of Theorem 8. The dotted lines indicate the position of agent xi before moving her from the left side to the right side of y. The cost of every agent with respect to y is the same before and after moving her to the right. The ﬁgure depicts only three agents (including x1 and xn) where SCM2(x) is the social cost of Mechanism M2 on instance x and SCy(x) is the social cost admitted by y on instance x. By the way the instance was constructed, it holds that SCM2(x) ≥SCM2(x′) and SCy(x) = SCy(x′) = SCopt(x′) and hence SCM2(x)/SCy(x) is an upper bound on SCM2(x′)/SCopt(x′). p Let di = (xi −b)−y, for i = 2, . . . , n and let k = i̸=1 di. Finally let d = \|y−(x1+b)\|. We consider three cases. (See Fig. 3). ( g ) se 1 x1 + b < y. Case 1 x1 + b < y. Case 1 x1 + b < y. In this case, the social cost admitted by y is SCy(x) = nc + k + d, and the cost of Mechanism M2 is SCM2(x) = nc + k + (n −1)(d + 2b). The ratio is SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(d + 2b) + nc k + d + nc . cost(y, xi), is not strategyproof. So the ratio is In this case, the social cost admitted by y is SCy(x) = k + d + nc, and the cost of Mechanism M2 is SCM2(x) = k + (n −1)(2b −d) + nc. So the ratio is SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(2b −d) + nc k + d + nc . SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(2b −d) + nc k + d + nc . Again, we calculate the partial derivative of g(k, d, n) with respect to k, ∂g(k, d, n) ∂k = k + d + nc −k −(n −1)(2b −d) −nc (k + d + nc)2 = d −(n −1)(2b −d) (k + d + nc)2 . Since 0 ≤d < b, d < 2b −d and the above value is negative. To get the maximum value of the ratio, we plug in k = 0 and the approximation ratio becomes Since 0 ≤d < b, d < 2b −d and the above value is negative. To get the maximum value of the ratio, we plug in k = 0 and the approximation ratio becomes SCM2(x) SCy(x) = g(d, n) = (n −1)(2b −d) + nc d + nc . By calculating the partial derivative with respect to d, we get ∂g(d, n) ∂d = −(n −1)(d + nc) −(n −1)(2b −d) −nc (d + nc)2 = −(n −2)nc −(n −1)2b (d + nc)2 < 0. So the ratio is maximized when d = 0, and the ratio is 2b(n−1)+nc nc < 1 + 2b c . Case 3 x1 b < y ≤x1 which means b ≤d < 2b Case 3 x1 −b < y ≤x1, which means b ≤d < 2b. 1 y ≤ 1, ≤ In this case, the social cost admitted by y is SCy(x) = k + 2b −d + nc, and cost of Mechanism M2 is SCM2(x) = k + (n −1)(2b −d) + nc. So the ratio is SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(2b −d) + nc k + 2b −d + nc . Mechanism M2 is SCM2(x) = k + (n −1)(2b −d) + nc. So the ratio is SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(2b −d) + nc k + 2b −d + nc . cost(y, xi), is not strategyproof. By calculating the partial derivative of the ratio with respect to k, we have By calculating the partial derivative of the ratio with respect to k, we have ∂g(k, d, n) ∂k = k + d + nc −k −(n −1)(d + 2b) −nc (k + d + nc)2 = d −(n −1)(d + 2b) (k + d + nc)2 < 0. When k = 0, i.e. xi = y + b for all agents i = 2, . . . , n, the ratio achieves the maximum value with respect to k. We plug in k = 0 to the above formula and get SCM2(x) SCy(x) = g(d, n) = (n −1)(d + 2b) + nc d + nc = (n −1)d + 2(n −1)b + nc d + nc . 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1226 Again, we calculate the partial derivative of g(d, n) with respect to d, Again, we calculate the partial derivative of g(d, n) with respect to d, ∂g(d, n) ∂d = (n −1)(d + nc) −(n −1)d −2(n −1)b −nc (d + nc)2 = c(n2 −4n + 2) (d + nc)2 . For sufﬁciently large n (n ≥4), the above value is positive, so the ratio is maximized when d approaches to inﬁnity, which means that agent x1 is positioned very far away from the rest of the agents coinciding on position xi. Therefore the ratio goes to n −1. For sufﬁciently large n (n ≥4), the above value is positive, so the ratio is maximized when d approaches to inﬁnity, which means that agent x1 is positioned very far away from the rest of the agents coinciding on position xi. Therefore the ratio goes to n −1. Case 2 x1 < y ≤x1 + b, which means 0 ≤d < b. In this case, the social cost admitted by y is SCy(x) = k + d + nc, and the Mechanism M2 is SCM2(x) = k + (n −1)(2b −d) + nc. So the ratio is SCM2(x) SCy(x) = g(k, d, n) = k + (n −1)(2b −d) + nc k + d + nc . In this case, the social cost admitted by y is SCy(x) = k + d + nc, and Mechanism M2 is SCM2(x) = k + (n −1)(2b −d) + nc. cost(y, xi), is not strategyproof. The main intuition behind the proof of the lemma is that we will simulate proﬁles with two agents with proﬁles with n agents, where two groups of n/2 agents coincide on two different positions. The two-agent mechanism will then output the same location that the n-agent mechanism outputs on the respective location proﬁle. Similar ideas have been used before in the literature, e.g. see [19,36]. Lemma 7 Let Mn be a strategyproof, anonymous and position invariant mechanism for n agents, where n is even. Then, for any location proﬁle x = ⟨x1 = · · · = xn/2, xn/2+1 = · · · = xn⟩, it holds that Mn(x) = x1 −b. Proof Let M2 be the following mechanism for two agents: On input location proﬁle ⟨x1, x2⟩, output Mn(x′), where x′ = ⟨x′ 1 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩, and x′ 1 = x1 and x′ n/2+1 = x2. First, we claim that M2 is strategyproof, anonymous and position invariant. If that is true, then by Theorem 6, M2 is Mechanism M2 and the lemma follows. Proof Let M2 be the following mechanism for two agents: On input location proﬁle ⟨x1, x2⟩, output Mn(x′), where x′ = ⟨x′ 1 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩, and x′ 1 = x1 and x′ n/2+1 = x2. First, we claim that M2 is strategyproof, anonymous and position invariant. If that is true, then by Theorem 6, M2 is Mechanism M2 and the lemma follows. First let x = ⟨x1, x2⟩, ˆx = ⟨ˆx1, ˆx2⟩be any two position equivalent location proﬁles. Observe that the corresponding n-agent proﬁles x′ and ˆx′ obtained by placing n/2 agents on x1 and ˆx1 and n/2 agents on x2 and ˆx2 respectively are also position equivalent. Since Mn is position invariant, it must hold that Mn(x′) = Mn(ˆx′) and hence by construction of M2, M2(x) = M2(ˆx). Since x and ˆx where arbitrary, Mechanism M2 is position invariant. Similarly, let x = ⟨x1, x2⟩, ˆx = ⟨ˆx1, ˆx2⟩be any two location proﬁles, such that ˆx is obtained by x by a permutation of the agents. cost(y, xi), is not strategyproof. We ﬁrst calculate partial derivative of the function g with respect to k, We ﬁrst calculate partial derivative of the function g with respect to k, ∂g(k, d, n) ∂k = k + 2b −d + nc −k −(n −1)(2b −d) −nc (k + 2b −d + nc)2 = (2 −n)(2b −d) (k + 2b −d + nc)2 < 0. The function is decreasing in k so let’s set k = 0. The function f becomes The function is decreasing in k so let’s set k = 0. The function f becomes SCM2(x) SCy(x) = g(d, n) = (n −1)(2b −d) + nc 2b −d + nc . Now, we calculate partial derivative of g with respect to d and get ∂g(d, n) ∂d = −(n −1)(2b −d) −n(n −1)c + (n −1)(2b −d) + nc (2b −d + nc)2 ≤0. 123 1227 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 So function g(d, n) is decreasing in d. Let’s set d = b, and the ratio goes to (n−1)b+nc b+nc , which is always smaller than n −1. In all, since SCM2(x)/SCy(x) ≤n −1 the approximation ratio of Mechanism M2 is at most max{n −1, 1 + 2b c }. The approximation ratio is exactly n −1 on any instance x = ⟨x1, x2, . . . , xn⟩with x2 = · · · = xn and x1 ≪x2, i.e. when agent 1 lies on the left, really far away from the other n −1 agents. We note that by our analysis, it follows that the other upper bound of the ratio of the mechanism is actually 1 + (n−1)2b nc but we instead wrote a larger bound of 1 + 2b c for ease of exposition. The ratio is exactly 1 + (n−1)2b nc on any instance x = ⟨x1, x2, . . . , xn⟩with x1 + b = x2 −b = · · · = xn −b and goes to 1 + 2b c as n goes to inﬁnity. ⊓⊔ ⊓⊔ Next, we will prove a lower bound of 1+b/c on the approximation ratio of any anonymous, position invariant, and strategyproof mechanism, when the number of agents is even. We will start with the following lemma. cost(y, xi), is not strategyproof. In fact, as we will p Auton Agent Multi-Agent S ﬁle ˆx = ⟨ˆx1, ˆx′ 2⟩and by e the choice of x1 (and h f. ven, no strategyproof me e an approximation ratio and position invariant me /2 + 1 = · · · = xn with t of Mn is nc + (n/2) · 2 r bound follows. tputsthelocationthatmi x2⟩with x1 + b < x2 − reports x′ 2 = 2x2 −2b − nism M2 for the social . In fact, as we will pro hi h b ibl Auton Agent ation proﬁle ˆx = ⟨ˆx1 and since the choice ategyproof. gents is even, no strate an achieve an approxi onymous and position = xn/2, xn/2 + 1 = · · · ocial cost of Mn is nc The lower bound follo smthatoutputstheloc ance ⟨x1, x2⟩with x1 nt x2 misreports x′ 2 = st. of Mechanism M2 fo d the case. In fact, as actually achieves the b 1228 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 x′ n⟩corresponds to the 2-agent location proﬁle ˆx = ⟨ˆx1, ˆx′ 2⟩and by construction of M2, cost(ˆx′ 1, M2(ˆx′)) ≤cost(ˆx′ 1, M2(ˆx)) and since the choice of x1 (and hence the choice of ˆx1) was arbitrary, Mechanism M2 is strategyproof. ⊓⊔ Theorem 9 When the number of agents is even, no strategyproof mechanism that satisﬁes position invariance and anonymity can achieve an approximation ratio lower than 1+(b/c) for the social cost. Proof Let Mn be a strategyproof, anonymous and position invariant mechanism and consider any location proﬁle x = ⟨x1 = · · · = xn/2, xn/2 + 1 = · · · = xn with xn/2+1 = x1 + 2b. By Lemma 7, Mn(x) = x1 −b and the social cost of Mn is nc + (n/2) · 2b while the social cost of the optimal allocation is only nc. The lower bound follows. ⊓⊔ 4.2 Maximum cost First,itiseasytoseethatthemechanismthatoutputsthelocationthatminimizesthemaximum cost is not strategyproof. On any instance ⟨x1, x2⟩with x1 + b < x2 −b the optimal location of the facility is (x1 + x2)/2. If agent x2 misreports x′ 2 = 2x2 −2b −x1 then the location moves to x2 −b, minimizing her cost. While the approximation ratio of Mechanism M2 for the social cost is not constant, for the maximum cost that is indeed the case. In fact, as we will prove, when the number of agents is even, Mechanism M2 actually achieves the best possible approximation ratio amongst strategyproof mechanisms. We start with the theorem about the approximation ratio of Mechanism M2. Theorem 10 For n ≥3, Mechanism M2 achieves an approximation ratio of 1 + 2b c for the maximum cost. Proof Let x = ⟨x1, . . . , xn⟩be any instance. We consider three cases, depending on the distance between agents x1 and xn. Case 1 x1 + b ≤xn −b ⇒xn −x1 ≥2b. Case 1 x1 + b ≤xn −b ⇒xn −x1 ≥2b. In this case, the cost of the optimal mechanism is MCopt(x) = (xn −b) −x1+xn 2 + c = xn−x1 2 −b + c, whereas the cost of Mechanism M2 is MCM2(x) = xn −x1 + c. The approximation ratio is MCM2(x) MCopt(x) = 2(xn −x1 + c) xn −x1 −2b + 2c = 2 + 4b −2c xn −x1 −2b + 2c ≤1 + 2b c . Hence, in this case, the approximation ratio is at most 1 + 2b c . For xn −b = x1 + b (the instance on which the right peak of the ﬁrst agent and the left peak of the last agent coincide), the approximation ratio is exactly 1 + 2b c . Hence, in this case, the approximation ratio is at most 1 + 2b c . For xn −b = x1 + b (the instance on which the right peak of the ﬁrst agent and the left peak of the last agent coincide), the approximation ratio is exactly 1 + 2b c . Case 2 x1 < xn −b < x1 + b ⇒c < xn −x1 < 2b. The cost of Mechanism M2 in this case is MCM2(x) = xn −b−(x1−b)+c = xn −x1+c, ile the cost of the optimal mechanism is at least c. cost(y, xi), is not strategyproof. The outcome of Mn on the corresponding n-agent location proﬁles (since the number of agents placed on x1 and x2 is the same) is the same and by construction of M2, M2(x) = M2(ˆx) and since the proﬁles where arbitrary, the mechanism is anonymous. Finally, for strategyproofness, start with a location proﬁle ˆx′ = ⟨ˆx′ 1, ˆx′ 2⟩and let x′ = ⟨x′ 1 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩be the corresponding n-agent location proﬁle. Let y = Mn(x′) and let cost(x′, y) be the cost of agents x′ 1, . . . , x′ n/2 on x′. For any x1, let ⟨x1, x′ 2 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩be the resulting location proﬁle. By strategyproofness of Mn, agent x′ 1 can not decrease her cost by misreporting x1 on proﬁle x′ and hence her cost on the new proﬁle is at least cost(x′, y). Next, consider the location proﬁle ⟨x1 = x2, x′ 3 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩and observe that by the same argument, the cost of agent x′ 2 is not smaller on the new proﬁle when compared to ⟨x1, x′ 2 = · · · = x′ n/2, x′ n/2+1 = · · · = x′ n⟩and hence her cost is at least cost(x′, y). Continuing like this, we obtain the proﬁle ⟨x1 = · · · = xn/2, x′ n/2+1 = · · · = x′ n⟩and by the same argument, the cost of agent x′ n/2 on this proﬁle is at least cost(x′, y). The location proﬁle ⟨x1 = · · · = xn/2, x′ n/2+1 = · · · = Auton Agent Multi-Agen on proﬁle ˆx = ⟨ˆx1, ˆx′ 2⟩and b nd since the choice of x1 (and gyproof. nts is even, no strategyproof m achieve an approximation ra ymous and position invariant m n/2, xn/2 + 1 = · · · = xn with ial cost of Mn is nc + (n/2) · he lower bound follows. mthatoutputsthelocationthatm ce ⟨x1, x2⟩with x1 + b < x2 − x2 misreports x′ 2 = 2x2 −2b Mechanism M2 for the soci he case. 4.2 Maximum cost The approximation ratio is The cost of Mechanism M2 in this case is MCM2(x) = xn −b−(x1−b)+c = xn −x1+c, while the cost of the optimal mechanism is at least c. The approximation ratio is MCM2(x) MCopt(x) ≤(xn −x1 + c) c < 1 + 2b c Case 3 x1 −b ≤xn −b ≤x1 ⇒xn −x1 ≤b. 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1229 The cost of Mechanism M2 is MCM2(x) = xn −x1 + c while the cost of the optimal mechanism is at least c. The approximation ratio is MCM2(x) MCopt(x) ≤(xn −x1 + c) c ≤1 + b c . MCM2(x) MCopt(x) ≤(xn −x1 + c) c ≤1 + b c . Over the three cases, the worst approximation ratio is at most 1 + 2b c and there is actually an instance of the problem with approximation ratio exactly 1 + 2b c , ensuring that the bound is tight. ⊓⊔ Over the three cases, the worst approximation ratio is at most 1 + 2b c and there is actually an instance of the problem with approximation ratio exactly 1 + 2b c , ensuring that the bound is tight. ⊓⊔ number of agents is even f even, no deterministic str ymity can achieve an app em 9, there are agents wh t mechanism is 2b + c, w n the approximation ratio number of agents (and wi eorem. f mechanism can achieve th x1 +b < x2 −b (the in on positions x1 and x2 and ty is fopt(x) = x1+x2 2 . As hanism with approximatio The lower bound for the case when the number of agents is even follows. umber of agents is even even, no deterministic s mity can achieve an ap m 9, there are agents w mechanism is 2b + c, the approximation ratio number of agents (and w eorem. mechanism can achieve h x1 +b < x2 −b (the i on positions x1 and x2 an y is fopt(x) = x1+x2 2 . A anism with approximat ∈[x2−b, ∞). Let d = −b, ∞), then it would b t 2 − c d+c which goes to each other). 4.2 Maximum cost Now, for M be fM(x) ∈[ fopt(x) x Corollary 1 When the number of agents is even, no deterministic strategyproof mechanism that satisﬁes position invariance and anonymity can achieve an approximation ratio lower than 1 + 2b c for the maximum cost. Proof On instance ˆx of the proof of Theorem 9, there are agents whose cost for any strat- egyproof, anonymous and position invariant mechanism is 2b + c, while under the optimal mechanism it is only c. The lower bound on the approximation ratio follows. ⊓⊔ A different lower bound that holds for any number of agents (and without using the position invariance property) is proved in the next theorem. Theorem 11 No deterministic strategyproof mechanism can achieve an approximation ratio lower than 2 for the maximum cost. Theorem 11 No deterministic strategyproof mechanism can achieve an approximation ratio lower than 2 for the maximum cost. Proof Consider an instance x = ⟨x1, x2⟩with x1 +b < x2 −b (the instance can be extended to arbitrarily many agents by placing agents on positions x1 and x2 and all the arguments will still hold). The optimal location of the facility is fopt(x) = x1+x2 2 . Assume for contradiction that M is a deterministic strategyproof mechanism with approximation ratio smaller than 2. First, we argue that it can not be that fM(x) ∈[x2−b, ∞). Let d = x2−b−fopt(x). It holds that MCopt(x) = c+d. If it was fM(x) ∈[x2−b, ∞), then it would be that MCM(x) ≥c+2d and the approximation ratio would be at least 2 − c d+c which goes to 2 as d grows to inﬁnity (i.e. the agents are placed very far away from each other). Now, for Mechanism M to achieve an approximation ratio smaller than 2, it must be fM(x) ∈[ fopt(x), x2−b) (or symmetrically fM(x) ∈(x1 + b, fopt(x)]). Now consider the instance x′ = ⟨x′ 1, x′ 2⟩with x′ 1 = x1 and x′ 2 = fM(x) + b. On this instance, it must be either fM(x′) = fM(x) or fM(x′) = fM(x) + 2b (the left or the right peak of agent x′ 2), otherwise agent x′ 2 could report x2 and move the facility to x′ 2 −b, minimizing her cost and violating strategyproofness. For calculating the lower bound, we need the choice that admits the smaller of the two costs, i.e. fM(x′) = fM(x). We calculate the approximation ratio on instance x′. The optimal choice for the facility is again fopt(x′) = (x1 + x′ 2)/2. Let λ = (x′ 2 −b) − fopt(x′). The approximation ratio then is 2 − c λ+c. We know that λ ≥d/2, so when d grows to inﬁnity as before, λ also grows to inﬁnity and the approximation ratio goes to 2. This means that there exists an instance for which the approximation ratio of the mechanism is 2, which gives us the lower bound. ⊓⊔ 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1230 5 Group strategyproofness , xn⟩, where x1 = · · · = xk+1, xk+2 = · · · = xn and (xn −b) −(x1 + b) = 2b. Similarly, any position invariant and group strategyproof mechanism that outputs f (xS) = xS n −b on instance S, must output f (x) = xn −b on any instance x = ⟨x1, . . . , xn⟩, where x1 = · · · = xk, xk+1 = · · · = xn and (xn −b) −(x1 + b) = 2b. Proof We prove the ﬁrst part of the lemma. The proof of the second part is symmetric. Note that the difference between instances xP and x is that the distance between the two groups of agents is 3b+ϵ in xP while it is 4b in x. First, we argue that f (x) ∈[x1+b, xn −b]. Indeed, if that was not the case, if f (x) < x1 +b, by the onto condition implied by position invariance, all agents could jointly misreport some different positions and move the facility to x1 + b. This point admits a smaller cost for all agents; speciﬁcally the cost of agents x1, . . . , xk+1 is minimized while the cost of agents xk+2, . . . , xn is reduced and group strategyproofness is violated. Using a symmetric argument, we conclude that it can’t be f (x) > xn −b either. Secondly, we argue that f (x) /∈(x1 + b, xn −b]. Indeed, assume that was not the case. Then agents xP k+2, . . . , xP n on instance xP could jointly misreport xk+2, . . . , xn and move the facility from f (xP) = xP 1 +b to f (x). Since by assumption f (x) ∈(x1 +b, xn −b], the cost of each deviating agent is smaller than her cost before deviating. Group strategyproofness is then violated and hence it must be that f (x) = x1 + b. By position invariance, it must be that f (¯x) = ¯x1 + b on any instance ¯x which is position equivalent to instance x. ⊓⊔ Proof We prove the ﬁrst part of the lemma. The proof of the second part is symmetric. Note that the difference between instances xP and x is that the distance between the two groups of agents is 3b+ϵ in xP while it is 4b in x. 5 Group strategyproofness As we mentioned in the introduction, under the reasonable conditions of position invariance and anonymity, there is no group strategyproof mechanism for the problem. We will prove this claim by using Lemma 4 and the following lemma. Recall the deﬁnition of the primary instance from Sect. 4. When n = 2k + 1, k ∈Z+, let P be the instance obtained by locating k + 1 agents on x1 and k agents on x2 on the primary instance. Similarly, let S be the instance obtained by locating k agents on x1 and k +1 agents on x2 on the primary instance. Formally, let xP = ⟨x P 1 , . . . , x P n ⟩, where x P 1 = · · · = x P k+1, x P k+2 = · · · = x P n , and (x P n −b) −(x P 1 + b) = b + ϵ and xS = ⟨x S 1 , . . . , x S n ⟩, where x S 1 = · · · = x S k , x S k+1 = · · · = x S n , and (x S n −b) −(x S 1 + b) = b + ϵ, where ϵ is the same quantity as in the primary instance. Lemma 8 When n = 2k + 1, any position invariant and group strategyproof mechanism that outputs f (xP) = xP 1 + b on instance P, must output f (x) = x1 + b on any instance x = ⟨x1, . . . , xn⟩, where x1 = · · · = xk+1, xk+2 = · · · = xn and (xn −b) −(x1 + b) = 2b. Similarly, any position invariant and group strategyproof mechanism that outputs f (xS) = xS n −b on instance S, must output f (x) = xn −b on any instance x = ⟨x1, . . . , xn⟩, where x1 = · · · = xk, xk+1 = · · · = xn and (xn −b) −(x1 + b) = 2b. Lemma 8 When n = 2k + 1, any position invariant and group strategyproof mechanism that outputs f (xP) = xP 1 + b on instance P, must output f (x) = x1 + b on any instance x = ⟨x1, . . . 5 Group strategyproofness First, we argue that f (x) ∈[x1+b, xn −b]. Indeed, if that was not the case, if f (x) < x1 +b, by the onto condition implied by position invariance, all agents could jointly misreport some different positions and move the facility to x1 + b. This point admits a smaller cost for all agents; speciﬁcally the cost of agents x1, . . . , xk+1 is minimized while the cost of agents xk+2, . . . , xn is reduced and group strategyproofness is i l d U i i l d h i ’ b f ( ) b i h Secondly, we argue that f (x) /∈(x1 + b, xn −b]. Indeed, assume that was not the case. Then agents xP k+2, . . . , xP n on instance xP could jointly misreport xk+2, . . . , xn and move the facility from f (xP) = xP 1 +b to f (x). Since by assumption f (x) ∈(x1 +b, xn −b], the cost of each deviating agent is smaller than her cost before deviating. Group strategyproofness is then violated and hence it must be that f (x) = x1 + b. By position invariance, it must be that f (¯x) = ¯x1 + b on any instance ¯x which is position equivalent to instance x. ⊓⊔ ⊓⊔ Theorem 12 There is no group strategyproof mechanism that is anonymous and position invariant. Theorem 12 There is no group strategyproof mechanism that is anonymous and position invariant. Hence, it must be that either f (xP) = x P 1 +b or f (xP) = x P n −b. Assume w.l.o.g. that f (xP) = x P 1 + b; the other case can be handled symmetrically. 1 Following the arguments of case 2 of Lemma 4 (using instance PIII instead of III), group strategyproofness and position invariance imply that f (xPIII) = x PIII 1 −b or f (xPIII) = P Following the arguments of case 2 of Lemma 4 (using instance PIII instead of III), group strategyproofness and position invariance imply that f (xPIII) = x PIII 1 −b or f (xPIII) = x PIII 1 + b on instance PIII. By the arguments of subcase (a) (using instance PIVa instead of IVa), it can’t be that f (xPIII) = x PIII 1 −b, so it must be that f (xPIII) = x PIII 1 + b. However, we can not simply apply the argument used in subcase (b) to get a contradiction, because since there is a different number of agents on x P 1 and x P 2 , instances P and PIVa are no longer position equivalent. S Next, consider instance S and observe that f (xS) /∈(x S 1 +b, x S n −b) by the same arguments as above and f (xS) ∈[x S 1 + b, x S n −b] by group strategyproofness and position invariance. Hence it is either f (xS) = x S 1 + b or f (xS) = x S n −b. Assume ﬁrst that f (xS) = x S 1 + b. By the same arguments as above (using instances SIII and SIVa), on instance SIII, it must be that f (xSIII) = x SIII 1 + b. Now, observe that if X PIII 2 misreport ¯x PIII i = x PIII i −2b −2ϵ, then we get instance PIVb which is position equivalent to instance SIII and hence it must be that f (xPIVb) = x PIVb n + b. The cost of X PIII 2 before misreporting was b + c −ϵ while it becomes c + 2ϵ after misreporting. This violates strategyproofness, which means that on instance S, it must be f (xS) = x S n −b. n Let’s denote instance T by xT = ⟨xT 1 , . . . Theorem 12 There is no group strategyproof mechanism that is anonymous and position invariant. Proof When n = 2, on the primary instance, according to Lemma 4, there is no anonymous, position invariant and strategyproof mechanism such that f (x) ∈[x1+b, x2−b]. In addition, if the facility was placed on some point f (x) < x1 + b (the argument for f (x) > x2 −b is symmetric), for any mechanism that satisﬁes position invariance which implies onto, agents 1 and 2 could jointly misreport some positions ˆx1 and ˆx2 such that f (ˆx1, ˆx2) = x1 + b. Obviously, this deviation admits the minimum possible cost for agent 1 and a reduced cost for agent 2, violating group strategyproofness. The proof can easily be extended to the case when n is even. On the primary instance, simply place n 2 agents on x1 and n 2 agents on x2. By considering deviations of coalitions of agents coinciding on x1 or x2 instead of deviations of agents x1 and x2 respectively, all the arguments still hold. However, additional care must be taken when n is odd. 123 1231 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 When n = 2k + 1, k ∈Z+, we denote by PJ the instance after placing k + 1 agents on xJ 1 and k agents on xJ 2 on instance J, where J is either instance I, II, III, IVa or IVb of the proof of Lemma 4. Similarly, let SJ be the instance after placing k agents on xJ 1 and k + 1 agents on xJ 2 on instance J. Finally, let XK 1 be the group of agents i for which xK i = xK 1 on instance K, where K is either P, PI, PII, PIII, PIVa, PIVb, S, SI, SII, SIII, SIVa or SIVb, and let XK 2 be the group of agents i for which xK i = xK n on instance K. i For contradiction, assume that there exists an anonymous, position invariant, and group strategyproof mechanism. On instance P, by group strategyproofness, it must be f (xP) ∈ [x P 1 + b, x P n −b] and by the same arguments used in case 1 of Lemma 4 (with instances PI, PII instead of I, II and with X P 1 and X P 2 instead of x1 and x2)3, it must be f (xP) /∈ (x P 1 +b, x P n −b). 3 Here we assume for convenience that any mechanism outputs the same location in [x P 1 + c, x Pn −c] on the primary instance and instance P. This is without loss of generality because the argument for any output in [x P 1 + b, x Pn −b] is exactly the same. Theorem 12 There is no group strategyproof mechanism that is anonymous and position invariant. 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1232 6 Conclusion and discussion In this paper, we studied a natural variant of a well-known problem, that of truthful facility location, when agents have double-peaked linear preferences over the set of possible loca- tions. As we saw, the fact that double-peaked preferences are not as well-behaved as their well-studied single-peaked counterpart makes the problem of designing good truthful mech- anisms quite more challenging. Given the standard interpretation of the median agent as a majority outcome in the single-peaked domain, our results seem to indicate that even slightly more complicated preference structures (that do not admit such majorities) might necessarily have very bad performance guarantees in the absence of such a consensus and that some sort of randomization is in fact essential. Our work can be placed directly in the middle of the extensive literature on truthful facility location that comprises of many interesting settings, modelling different situations. The original model [1,30] assumes single-peaked preferences, the obnoxious facility model [7,8,16] assumes singled-dipped (also known as single-caved preferences) whereas the dual preference model [33,34,39] assumes a combination of single-peaked and single-dipped preferences. Each preference structure in these works is motivated by corresponding real-life scenarios; in this light, our model can also be seen as another interesting preference structure, motivated by a different realistic scenario, which immediately places it in tight connection to the related work in artiﬁcial intelligence and theoretical computer science. Furthermore, as we mentioned in the introduction, the interest in these types of problems is expanding with works considering different objectives or more complicated cost structures. Given that double-peaked preferences are a natural preference structure for some scenar- ios, as also advocated by some of the related work in economics, one could consider their use in other problems, beyond facility location. We have already mentioned their use in [38] for the problem of controlling elections; one could think of other uses in computational aspects of social choice, an important subﬁeld of artiﬁcial intelligence. For example, one could ask the questions of whether efﬁcient algorithms for deciding whether a given incomplete pref- erence structure can be extended to a double-peaked proﬁle, similarly to the corresponding questions for single-peaked preferences [3,15]. Another question is whether it is possible to efﬁciently elicit double-peaked preference orderings using some kind of query operation, e.g. comparison queries, like for instance in [9] for the single-peaked case. Theorem 12 There is no group strategyproof mechanism that is anonymous and position invariant. , xT n ⟩, where xT 1 = · · · = xT k = xT k+1 −2b = xT k+2 −4b = · · · = xT n −4b. Let XT 1 be the set of agents i for which xT i = xT 1 , and let X T 2 be the set of agents j for which xT j = xT n , and let xt be agent xT k+1. On instance T, it must be either f (xT ) = xT 1 + b or f (xT ) = xT n −b, otherwise agent xt could misreport x′ t = xT 1 and then by Lemma 8 and the fact that on instance P it is f (xP) = x P 1 + b, it should be f (xT 1 , . . . , x′ t, . . . , xT n ) = xt −b, which admits a cost of c for agent xt. Similarily, by Lemma 8 and the fact that on instance S, it is f (xS) = x S n −b, if agent xt misreports x′′ t = xT n , then it should be f (xT 1 , . . . , x′′ t , . . . , xT n ) = xt + b. If f (xT) = xT n −b, agent xt could form a coalition with agents XT 1 and by misreporting x′ t, move the facility to xt −b, a choice that would admit the same cost for her, but a strictly smaller cost for every other member of the coalition. If f (xT) = xT 1 + b then agent xt could form a coalition with agents XT 2 and by misreporting x′′ t , move the facility to xt + b, a choice that would admit the same cost for her, but a strictly smaller cost for every other member of the coalition. In each case, there is a coalition of agents that can beneﬁt from misreporting and group strategyproofness is violated. This completes the proof. ⊓⊔ ⊓⊔ 3 Here we assume for convenience that any mechanism outputs the same location in [x P 1 + c, x Pn −c] on the primary instance and instance P. This is without loss of generality because the argument for any output in [x P 1 + b, x Pn −b] is exactly the same. 7 Future work and extensions Starting from randomized mechanisms, we would like to obtain lower bounds that are func- tions of b and c, to see how well Mechanism M1 fares in the general setting. For deterministic mechanisms, we would like to get a result that would clear up the picture. Characterizing strategyproof, anonymous and position invariant mechanisms would be ideal, but proving a lower bound that depends on n on the ratio of such mechanisms (for the social cost) would also be quite helpful. The techniques used in our characterization for two agents and our lower bounds seem to convey promising intuition for achieving such a task. The problem could be extended to the case of multiple peaks, instead of two peaks, while at the same time moving to more general metric spaces. Perhaps the most natural extension would be a setting where each agent is associated with a position on a metric space, a “center” and her most preferred locations lie on a sphere centered around this position. Equivalently, one could think of each agent being associated with all the points on the sphere centered around her location and her cost would be the distance between the location of the facility 123 Auton Agent Multi-Agent Syst (2017) 31:1209–1235 1233 Table 2 The results for the case when peaks are not required to be symmetric Non-symmetric Ratio Lower Social cost Deterministic n −1 1 + b1+b2 c Randomized n −1 − Maximum cost Deterministic 1 + b1+b2 c 1 + b1+b2 c Randomized 1 + b1+b2 c 3/2 Table 2 The results for the case when peaks are not required to be symmetric and the closest such point on the sphere. Crucially though, the agent can only manipulate the position of these points throught misreporting the position of the center of the sphere. This extension is interesting becauses it models precisely the same motivating example we used in the introduction, in an even more realistic setting. For example, case of the two-dimensional space corresponds to a scenario where the government is planning to build a primary school at some spot in a neighbourhood or a city block, instead of simply a street; a city block is much better modelled as a plane, instead of a real line. Table 2 The results for the case when peaks are not required to be symmetric 7 Future work and extensions This extension is also in accordance with the usual questions posed in the literature of facility location, where the investigations usually start from the case of one facility and the line metric and extend to multiple facilities on general metric spaces. The extension to multiple facilities, even for the line metric is of course also meaningful for our model of double-peaked preferences. Adifferentextensioncouldbeintermsofthecostfunctionsusedinthemodel.Althoughthe symmetric case is arguably the best analogue of the single-peaked preference setting, it could certainly make sense to consider a more general model, where the cost functions do not have the same slope in every interval and hence the peaks are not equidistant from the location of an agent. Let b1 and b2 be the distances from the left and the right peaks respectively. Clearly, all our lower bounds still hold, although one could potentially prove even stronger bounds by taking advantage of the more general setting. The main observation is that Mechanism M1 is no longer truthful-in-expectation, because its truthfulness depends heavily on the peaks being equidistant. On the other hand, mechanism M2 is still strategyproof and the approximation ratio bounds extend naturally. A summary of the results for the non-symmetric setting is depicted in Table 2. Future work could also consider a different choice for the objective function. Here, we studied the objective functions of the social cost and the maximum cost, following the original literature on the problem with single-peaked preferences [30]. Since then, several other objectives functions have been considered in the literature such as the least-squares objective, [18], the L p norm of costs[17] or the minimax envy [5]; it would make sense to consider the same or at least similar objectives for the case of double-peaked preferences as well. Finally, it would be meaningful to consider the problem under the veriﬁcation framework [21,22], especially if it turns out that strong inapproximability bounds apply, at least for the case of deterministic strategyproof mechanisms. 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https://openalex.org/W4229006324	https://eprints.gla.ac.uk/270669/1/270669.pdf	English	null	GIS and urban data science	Annals of GIS	2,022	cc-by	3,245	Annals of GIS ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/tagi20 Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=tagi20 CONTACT Yijing Li yijing.li@kcl.ac.uk CUSP London, Department of Informatics, King’s College London, London WC2B 4BG, UK © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Nanjing Normal University. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT With the emergence of new forms of geospatial/urban big data and advanced spatial analytics and machine learning methods, new patterns and phenomena can be explored and discovered in our cities and societies. In this special issue, we presented an overview of nine studies to understand how to use urban data science and GIS in healthcare services, hospitality and safety, transportation and mobility, economy, urban planning, higher education, and natural disasters, spreading across developed countries in North America and Europe, as well as Global South areas in Asia and the Middle East. The embrace of diverse geo-computational methods in this special issue brings forward an outlook to future GIS and Urban Data Science towards more advanced computational capability, global vision and urban-focused research. Background Mulligan, 2007; Fotheringham, Brunsdon, and Charlton 2002), spatial autocorrelation and regression models (Anselin, 2009), spatio-temporal Bayesian modelling (Hu et al. 2018; Haining and Li 2020; Law, Quick, and Jadavji 2020), Risk Terrain Modelling (RTM) (Caplan, Kennedy and Mille 2011), etc., had been utilized widely in urban fields, such as healthcare accessibility (Kim, Byon, and Yeo 2018), services resilience especially against the pandemic (Li, 2021); urban safety and crime mapping (Hart, Lersch, and Chataway 2020), crime pat tern detection (Chainey, 2020) aggregated at street seg ments (Tom-Jack, Bernstein, and Loyola 2019) and urban mobility (Sinclair et al. 2021: Sarim et al., 2021), hence brings forward the essentiality to apply such computa tional techniques in a geographically holistic sense. The special issue of Annals of GIS on GIS and Urban Data Science had compiled nine novel and original manu scripts demonstrating research on the main aspects of geo-informatics applications in urban contexts, in the hope of drawing follow-up inspirations towards further contributions to the literature. In the information age (Castells 1998), new forms of urban big data demand an in-depth understanding and diversi fied interpretation of human interactions with contem porary society. Urban Informatics (Shi et al. 2021) has surged as an interdisciplinary subject encompassing urban data science, informatics, GIS, various information and communication technology, big data analytics, smart cities, etc., and encourage people from academics, indus try and public sectors to jointly explore the open data emerging in the context of urban environments (Foth, Choi, and Satchell 2011), such as social media data (Wang and Li 2021; Wang, Gerber, and Brown 2012; Chen, Cho, and Jang 2015), mobile data (Bogomolov et al. 2014; Ros’es, Kadar, and Malleson 2021), Geo- locational data (Kikuchi, Amemiya, and Shimada 2012; Hutt, Bowers, and Johnson 2021; Song et al. 2019; Tu et al. 2021), image data (Law et al., 2020; Woodworth et al. 2014; Liu et al. 2020) and video data (Ashby 2019; Lindegaard and Bernasco 2018). It has often been applied to visualize the digital information from the society we reside in, to interpret the human interactions with sur rounding urban socio-economic environments, and to provide data-driven evidence for decision-making and industrial innovation. GIS and urban data science Yijing Li a, Qunshan Zhao b and Chen Zhong c aCUSP London, Department of Informatics, King’s College London, London, UK; bUrban Big Data Centre, School of Social and Political Sciences, University of Glasgow, Glasgow, UK; cCentre for Advanced Spatial Analysis, University College London, London, UK KEYWORDS GIS; urban data science; urban analytics; informatics KEYWORDS GIS; urban data science; urban analytics; informatics Yijing Li, Qunshan Zhao & Chen Zhong To cite this article: Yijing Li, Qunshan Zhao & Chen Zhong (2022) GIS and urban data science, Annals of GIS, 28:2, 89-92, DOI: 10.1080/19475683.2022.2070969 To cite this article: Yijing Li, Qunshan Zhao & Chen Zhong (2022) GIS and urban data science, Annals of GIS, 28:2, 89-92, DOI: 10.1080/19475683.2022.2070969 To link to this article: https://doi.org/10.1080/19475683.2022.2070969 To link to this article: https://doi.org/10.1080/19475683.2022.2070969 © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Nanjing Normal University. Published online: 07 May 2022. Submit your article to this journal Article views: 706 View related articles View Crossmark data ANNALS OF GIS 2022, VOL. 28, NO. 2, 89–92 https://doi.org/10.1080/19475683.2022.2070969 ANNALS OF GIS 2022, VOL. 28, NO. 2, 89–92 Overview of this special issue The articles included in this issue make significant con tributions to the research in GIS and urban data science. These nine papers represent the diversity of urban data applications in different sectors, healthcare services (e.g. hospital services), hospitality and safety (e.g. Airbnb and crime), transportation and mobility (e.g. bike sharing), In parallel boom of 21st century’s development of computer science and data visualization, GIS, geocom putation and urban analytics research methods, such as Kernel Density Estimation (KDE) (Bailey and Gatrell, 1995), geographically weighted regression (Cahill and CONTACT Yijing Li 90 Y. LI ET AL. Y. LI ET AL. 90 economy (e.g. retail business and labour force), urban planning (e.g. nightlight data), higher education (e.g. big scholarly data) and natural disasters (e.g. winter storm) across broad regions in North America, Europe and Asia, as depicted in Table 1. In combination of administrative data (e.g. Census) with the emerging forms of data such as bike sharing, airbnb, tweets, or night time light images, rich methods had been deployed including traditional (spatial) statistical or optimization methods such as OLS regression, GWR, and MCLP, as well as some advanced methods from GeoAI such as spatial clustering, travel pattern recognition, and text mining. in various applications, e.g. transport, retail, social interac tions, using conventional methods, e.g. OLS regression; kernel density estimation and comparatively more recent computation methods, e.g. spatial network analysis and density-based spatial clustering methods. In pursuing further advancements in GIS and data, the next three sub- outlooks present a research agenda for achieving such objectives. Advanced computational methods A clearly pointed research direction is to build on the knowledge and skill around the different domains. We see how much GIS has been advanced by new computa tional methods and the ‘invasion’ of data science. This trend will continue, as the 5Vs of big data challenge the conventional GIS way of handling and analysing data. Moreover, domain knowledge becomes even more important to assist and solve real-world problems with the support of AI. Acknowledgements We want to thank the authors who contributed to this Special Issue, as well as the reviewers who provided the authors with comments and very constructive feedback. Dr Qunshan Zhao has received UK ESRC’s ongoing support for the Urban Big Data Centre (UBDC) (ES/L011921/1 and ES/S007105/1). Dr Chen Zhong has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement No 949670), and from ESRC under JPI Urban Europe/NSFC (grant No. ES/T000287/ 1). Foth, M., J. H. Choi, and C. Satchell. 2011. Urban Informatics. Conference on Computer Supported Cooperative Work (CSCW). New York, NY, United States: Association for Computing Machinery, 1–8. 10.1145/1958824.1958826. Fotheringham, A. S., C. Brunsdon, and M. Charlton. 2002. Geographically Weighted Regression: The Analysis of Spatially Varying Relationships. Wiley. Haining, R. P., and G. Li. 2020. Modelling Spatial and Spatial- temporal Data: A Bayesian Approach, 608. Boca Raton: CRC Press. Hart, T. C., K. M. Lersch, and M. Chataway. 2020. Space, Time, and Crime. Durham, North Carolina: Carolina Academic Press. Urban focused research As most of the world’s population is moving to cities (UN report, 2018), they also generate new urban challenges, e.g. social inequality, housing issues, ageing society, and health hazards. Data-driven methods have been pro posed and demonstrated in this special issue to further tackle these problems, but more research and exploration need to be investigated to generate practical solutions and policy implementations for different urban issues. Caplan, J., L. Kennedy, and J. Miller. 2011. “Risk Terrain Modeling: Brokering Criminological Theory and GIS Methods for Crime Forecasting.” Justice Quarterly 28: 360–381. doi:10.1080/ 07418825.2010.486037. Castells, M. 1998. End of Millennium, the Information Age: Economy, Society and Culture Vol. III. Cambridge, MA; Oxford, UK: Blackwell. Chainey, S. 2020. Understanding Crime: Analyzing the Geography of Crime. Redlands, California: ESRI Press. Chen, X., Y. Cho, and S. Y. Jang. 2015. “Crime Prediction Using Twitter Sentiment and Weather.” 2015 Systems and Information Engineering Design Symposium 24 April 2015. Charlottesville, Virginia, USA: 63–68, doi: 10.1109/ SIEDS.2015.7117012. Outlook to the future Computational methods and data sciences are transform ing the conventional GIS into a new comprehensive disci pline, with the notation of urban analytics, location intelligence, and more recently GeoAI (Janowicz et al. 2020; Kandt and Batty 2021). As showcased in the paper on this special issue, geospatial research has been applied Table 1. Publication summary of the special issue. Table 1. Publication summary of the special issue. Publication Topic Data Spatial scale Methods Wang & Wang (2022) Hospital services ●2011 State Inpatient Databases (SID) of Florida (2,392,066 records); ●ZIP Code Tabulation Areas (ZCTAs) County/State (Florida, US) ●Refined Dartmouth method for defining HSAs; ●Spatialized network community detection methods for defining HSAs (constrained Louvain method-ScLouvain, and spatially constrained Leiden method- ScLeiden); ●3 quality measures for HSAs: localization index (LI), geographic compactness, and balanced region size. Beairsto et al. (2021) Sharing economy ●Bike sharing data from Nextbike Glasgow (2018– 2019) ●Scottish Index of Multiple Deprivation 2020 ●Scotland 2011 Census ●OpenStreetMap City (Glasgow, UK) ●OLS regression ●Two-step floating catchment area (2SFCA) methodology ●Maximum covering location problem (MCLP) Hladík et al. (2021) Retail services ●Places data from Facebook Graph API (6,800) City (Brno, Czech Republic) ●Choropleth map ●Nearest Neighbour Index ●Kernel density analysis ●Normalized Difference Index Maldonado- Guzmán (2020) Crime and sharing economy ●Number of crimes against property and people in 2017 (55,911) ●Airbnb lodging (InsideAirbnb) ●Census data City (Barcelona, Spain) ●OLS regression ●GWR model with Monte Carlo non-stationarity test Wu & Huang (2022) Human mobility patterns and social segregation ●Geotagged tweets December 2013 to May 2015 ●ACS data from US Census Cities (50 in US) ●DBSCAN for spatial clustering ●Race-ethnicity-economic mosaic ●Movement pattern analysis (home-activity) Yang et al. (2022) Urban morphology and urban planning Luojia-1A night-time light data Cities (331 in China) ●urban centre development index (UCDI) Zuo et al. 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https://openalex.org/W4387676785	https://ojs.uniska-bjm.ac.id/index.php/ziraah/article/download/11806/5637	Indonesian	null	PENGARUH PEMBERIAN TEPUNG SINGKONG (Manihot utilissima Pohl.) FERMENTASI TERHADAP BOBOT KARKAS DAN ORGAN PENCERNAAN AYAM BROILER	Ziraa'ah : Majalah Ilmiah Pertanian	2,023	cc-by	4,892	(The Effect Of Fermentation Cassava Flour (Manihot utillisima Pohl) On Percentage Of Carcass Weight And Digestive Organs Of Broiler Chicken) Lisnawaty Silitonga¹, Satrio Wibowo², Iis Yuanita³, Siti Ma’rifah⁴, Nopa Putriani⁵ 1,2,3,4,5Program Studi Peternakan, Fakultas Pertanian Universitas Palangka Raya Kampus UPR Tunjung Nyaho Jalan Yos Sudarso Palangka Raya Kalimantan Tengah Penulis koresponden: siti.ma’rifah@pet.upr.ac.id Lisnawaty Silitonga¹, Satrio Wibowo², Iis Yuanita³, Siti Ma’rifah⁴, Nopa Putriani⁵ 1,2,3,4,5Program Studi Peternakan, Fakultas Pertanian Universitas Palangka Raya Kampus UPR Tunjung Nyaho Jalan Yos Sudarso Palangka Raya Kalimantan Tengah Penulis koresponden: siti.ma’rifah@pet.upr.ac.id Naskah Diterima : 28-06-2023 Naskah Disetujui : 24-09-2023 Naskah Diterbitkan: 07-10-2023 This is an open-access article under the CC-BY 4.0 License. Copyright © 2023 by authors ABSTRACT This research was conducted to determine the effect of fermented cassava flour on the percentage of digestive organs and abdominal fat of broiler chickens. This study used a completely randomized design (CRD). The treatments were fermented cassava flour in feed P0=basal ration with 5% unfermented cassava flour; P1 = basal ration with 3% fermented cassava flour; P2=basal ration with 6% fermented cassava flour and P3=basal ration with 9% fermented cassava flour. Given fermented cassava flour until 9% in the feed was not a significant difference in the percentage of digestive organs of broiler chickens. ABSTRAK Penelitian ini bertujuan untuk mengetahui pengaruh pemberian tepung singkong fermentasi terhadap persentase organ pencernaan dan lemak abdominal ayam broiler. Penelitian ini menggunakan rancangan acak lengkap (RAL). Perlakuan yakni berupa pemberian tepung singkong fermentasi dalam pakan (P0=Ransum basal dan 5% tepung singkong tanpa fermentasi; P1=Ransum basal dan 3% tepung singkong fermentasi; P2=Ransum basal dan 6% tepung singkong fermentasi; dan P3=Ransum basal dan 9% tepung singkong fermentasi). Pemberian tepung singkong fermentasi hingga 9% dalam pakan tidak memberikan perbedaan yang nyata terhadap persentase bobot organ pencernaan ayam broiler. 394 394 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 PENDAHULUAN dipelihara, manajemen pemeliharaan dan pakan. Terpenuhinya kebutuhan pakan baik kualitas maupun kuantitas sangat menentukan penampilan produksi ternak Keberhasilan produksi suatu usah peternakan sangat ditentukan oleh beberapa faktor diantaranya sifat genetis ternak yang 395 p-ISSN 1412-1468 e-ISSN 2355-3545 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 kadar protein paling tinggi yaitu 40,860% dan kadar air rendah 6,64%, terjadi peningkatan kadar protein tepung ubi kayu seiring dengan lamanya fermentasi. Kenaikan protein ini disebabkan karena kemampuan dari Saccharomyces cerevisae maupun Rhizopus oryzae untuk mensekresikan beberapa enzim ektraseluler (protein) ke dalam singkong selama proses fermentasi, atau berkembangnya Saccharomyces cerevisae maupun Rhizopus oryzae ke dalam singkong dalam bentuk protein sel tunggal selama proses fermentasi (Kurniati et al., 2012). yang dibudidayakan terutama produksi ayam pedaging. Sifat genetis ayam pedaging memiliki laju pertumbuhan dan perkembangan yang sangat cepat, karena itu produksi yang optimal hanya bisa diwujudkan apabila ayam memperoleh pakan yang berkualitas baik dan dalam jumlah yang cukup (Hardini, 2003). Pakan adalah salah satu faktor yang sangat penting untuk mencapai suatu keberhasilan produktivitas ayam pedaging secara optimal, oleh karena itu kuantitas dan kualitas pakan hendaknya selalu diperhatikan. Biaya pakan merupakan komponen biaya terbesar yang mencapai 60- 70% dari total biaya produksi ternak unggas (Septiani et al., 2016). Menurut Badan Pusat Statistik Kota Palangka Raya, Produksi singkong yang ada di Provinsi Kalimantan Tengah berjumlah 45.712 ton (BPS, 2015). Dengan adanya produksi singkong yang berlimpah tersebut, maka dapat memberikan peluang bagi terciptanya pakan ternak unggas yang berkualitas dan terjangkau harganya. Tujuan dilaksanakannya penelitian ini adalah untuk mengetahui serta mengevaluasi pengaruh pemberian tepung singkong fermentasi terhadap bobot karkas dan organ pencernaan ayam broiler (hati, proventikulus, ventrikulus, usus halus dan usus besar). Singkong (Manihot utilissima Pohl.) merupakan salah satu tanaman tropis yang paling berguna dan secara luas dimanfaatkan sebagai sumber kalori yang murah, namun singkong mengandung asam sianida (HCN) yang bersifat toksik, sehingga masalah penurunan kadar HCN menjadi perhatian utama dalam pemanfaatan ubi kayu (Kobawila et al., 2005; Adamafio et al., 2010). Perbaikan karakteristik mutu tepung singkong dapat dilakukan dengan beberapa cara, antara lain secara fisika (temperatur dan tekanan), kimiawi (hidrolisa asam atau basa), dan biologis (fermentasi). Hasil perbaikan mutu tepung singkong dikenal dengan nama tepung mocaf (modified cassava flour) meliputi kenaikan viskositas, gelatinisasi, daya rehidrasi, dan kemudahan melarut (Rusdjijati, 2010). Ragi merupakan zat yang menyebabkan fermentasi. PENDAHULUAN Mikroorganisme yang terdapat pada ragi tape adalah kapang (Amylomyces rouxii, Mucor sp., dan Rhizopus sp.), khamir (Saccharomycopsis fibuligera, Saccharomycopsis malanga, Pichia burtonii, Saccharomyces cerevisiae, Candida utilis) dan bakteri (Pediococcus sp., Bacillus sp.). Amri dan Pratiwi (2014), mengemukakan proses pembuatan mocaf yaitu dengan proses fermentasi menggunakan beberapa jenis ragi salah satunya yaitu ragi tape, mocaf dengan menggunakan ragi tape merupakan tepung mocaf terbaik dengan METODE PENELITIAN Penelitian ini dilakukan pada bulan Juni hingga Agustus 2022 di Kandang Percobaan Program Studi Peternakan, Jurusan Budidaya Pertanian, Fakultas Pertanian, Universitas Palangka Raya. Alat yang digunakan dalam penelitian ini adalah ember plastik, nampan, timbangan digital, metern kain, oven, mesin pellet, kandang, bola lampu, tempat pakan dan tempat minum, pisau, plastik sampel, kamera, alat tulis, blender, pisau dan ayakan. Bahan yang digunakan dalam penelitian ini adalah ayam broiler sebayak 60 ekor (non sexing) strain cobb, ragi tape dan tepung singkong. Pengamatan dilakukan dari umur 1 hari hingga umur 35 hari.Pemberian air minum dilakukan secara ad libitum, pemberian pakan yang dicampur dengan tepung 396 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 singkong fermentasi dan diberikan setiap hari sesuai dengan kebutuhan. dilakukan dengan mengiris/menggunting bagian perut ke samping kiri dan kanan sampai pada bagian dada depan. Bagian yang telah diiris dibuka sehingga organ dalam terlihat. Kemudian organ dalam dikeluarkan dan diukur bobot dan panjangnya. Perlakuan yang diberikan terdiri atas : Penelitian ini menggunakan rancanggan acak lengkap (RAL) dengan 4 perlakuan dan 5 ulangan, jadi ada 20 unit perlakuan dan setiap perlakuan terdiri dari 3 ekor ayam (Purwinarto, dkk, 2020). Pada saat pemotongan diambil 2 ekor ayam dari setiap ulangan, sehingga jumlah ayam yang dipotong 40 ekor. Pemotongan dilakukan pada bagian antar tulang kepala dan tulang atas. Bagian yang dipotong terdiri atas 4 saluran, yaitu vena jugularis, arteri karotidae, oesophagus dan trakea. Ayam didiamkan sekitar 2 menit hingga mati agar darah keluar dengan sempurna. Ayam kemudian dicelupkan kedalam air panas sekitar 30 detik untuk mempermudah proses pencabutan bulu (Sucipto, 2020). Pembedahan ayam P0 = Ransum yang mengandung 5% tepung singkong tanpa fermentasi P1 = Ransum yang mengandung 3% tepung singkong fermentasi P2 = Ransum yang mengandung 6% tepung singkong fermentasi P3 = Ransum yang mengandung 9% tepung singkong fermentasi Berikut susunan ransum basal ayam broiler umur 0-35 hari dan kandungan nutrisnya yang disajikan dalam tabel 1 dan 2. nan Ransum Basal Ayam Broiler Umur 1-21 Hari dan kandungan nutrisinya Tabel 1. Susunan Ransum Basal Ayam Broiler Umur 1-21 Hari dan kandungan nutrisinya Tabel 1. Susunan Ransum Basal Ayam Broiler Umur 1-21 Hari dan kandungan nutrisinya No. Bahan Pakan R₀ (%) R₁ (%) R₂ (%) R₃ (%) 1. Jagung 37 35 37 35 2. Dedak Halus 23 27 16 10 3. AmpasTahu 20 20 26 31 4. Tepung Ikan 15 15 15 15 5. Variabel Penelitian Pemotongan dan pengambilan sampel dilakukan pada ayam broiler dengan umur 35 hari. Sampel yang dipotong sebanyak 2 ekor setiap ulangan. Pemotongan broiler sesuai dengan petunjuk Rasyaf (2003) yakni sebelum dipotong broiler dipuasakan 8 jam (air minum tetap diberikan), pemotongan pada pangkal leher broiler dengan memutuskan saluran pernafasan (trachea), saluran makan (esofagus) dan dua urat lehernya (pembuluh darah di kanan dan kiri leher), darah dibiarkan keluar minimal 3 menit (posisi broiler lebih baik digantung untuk melancarkan pengeluaran darah), broiler dicelupkan ke dalam air panas dengan temperatur 60 oC selama 30 detik (pastikan broiler sudah benar-benar dalam keadaan mati), kemudian seluruh bulu yang ada pada bagian tubuh broiler dicabut sampai bersih. Pengeluaran jeroan dimulai dari pemisahan tembolok dan trachea, membuka rongga badan dengan membuat irisan menyamping antara kloaka dan tulang dada, leher dan kepala serta kaki dipotong untuk mendapatkan karkas kosong kemudian dilakukan penimbangan. Setelah itu ayam akan dibedah dan diambil organ bagian dalamnya. Sebelum organ bagian dalam ditimbang, organ bagian dalam pencernaan akan dibersihkan dari lemak yang melekat. Bagian kepala, leher dan ceker dipisahkan. 3. Persentase hati (%), diperoleh dari perbandingan bobot hati dengan bobot potong ayam (Witantra, 2011). 4. Persentase proventikulus (%), diperoleh dari perbandingan bobot proventrikulus dengan bobot potong ayam (Witantra, 2011). 5. Persentase ventrikulus (%), diperoleh dari perbandingan bobot ventrikulus dengan bobot potong ayam (Witantra, 2011). 6. Persentase usus halus (%), diperoleh dari perbandingan bobot usus halus dengan bobot potongingan bobot hati dengan bobot potong ayam (Witantra, 2011). 7. Persentase usus besar (%), diperoleh dari perbandingan bobot usus besar dengan bobot potong ayam (Witantra, 2011). 8. Panjang usus halus (cm). 9. Panjang usus besar (cm). METODE PENELITIAN Tepung Singkong 5 0 0 0 6. Tepung Singkong Fermentasi 0 3 6 9 Jumlah 100 100 100 100 Premix 1 1 1 1 Kandungan Nutrisi 1. Protein Kasar (%) 20,443 20,702 21,166 21,62 2. Energi Metabolisme (kkal/kg) 3.011,77 3.071,93 3.012,59 3.003,45 3. Lemak Kasar (%) 6,18 6,57 5,63 5,09 4. Serat Kasar (%) 6,59 7,00 6,54 6,51 Tabel 2. Susunan Ransum Basal Ayam Broiler Umur 22-35 Hari No. Bahan Pakan R₀ (%) R₁ (%) R₂ (%) R₃ (%) 1. Jagung 47 40 45 35,5 2. Dedak Halus 15 26 10 10 3. AmpasTahu 18 16 24 30,5 4. Tepung Ikan 15 15 15 15 5. Tepung Singkong 5 0 0 0 6. Tepung Singkong Fermentasi 0 3 6 9 Jumlah 100 100 100 100 Kandungan Nutrisi 1. Protein Kasar (%) 20,279 20,702 20,7 21,53 2. Energi Metabolisme (kkal/kg) 3.009,05 3.020,61 3.062,35 3.000,5 3. Lemak Kasar (%) 5,4815 6,79 5,11 5,08 4. Serat Kasar (%) 5,4509 6,7 5,68 6,45 Tabel 2. Susunan Ransum Basal Ayam Broiler Umur 22-35 Hari 397 p-ISSN 1412-1468 e-ISSN 2355-3545 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 Analisis Data Penimbangan yang dilakukan meliputi karkas ayam, hati, proventrikulus, Data hasil penelitian dicatat dan ditabulasi dengan menggunakan program Microsoft Excel. Data bobot akhir, bobot karkas, persentase hati, persentase proventrikulus, persentase ampela, persentase usus halus, persentase usus besar, panjang usus halus dan usus besar dianalisis dengan analisis ragam uji F pada taraf α = 5% dan α = 1%. Apabila perlakuan berbeda sangat nyata, maka dilanjutkan dengan uji jarak berbeda Duncan (Duncan’s multiple range test) (Steel dan Torrie, 1995). ventrikulus/gizzard, usus halus dan usus besar serta dilakukan pengukuran panjang usus halus dan usus besar. Variabel yang diamati dalam penelitian ini adalah; Variabel yang diamati dalam penelitian ini adalah; 1. Bobot potong (g), diperoleh dari menimbang bobot ayam umur 35 hari setelah dipuasakan 8 jam (Sembiring, 2001). 2. Bobot karkas (g), diperoleh dari menimbang ayam pasca penyembelihan tanpa kepala, kaki dan jeroan (Sembiring, 2001) Bobot Potong dan Bobot Karkas Ayam Broiler Bobot Potong dan Bobot Karkas Ayam Broiler Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap bobot potong dan karkas ayam broiler dapat dilihat pada Tabel 3. 398 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 Tabel 3. Rataan bobot potong dan bobot karkas ayam broiler umur 35 hari Perlakuan Rata-Rata Bobot Potong (g) Karkas Bobot (g) Persentase (%) P0 (5%) 668,2a 411,6 62,14a P1 (3%) 690,3a 358,7 52,19a P2 (6%) 658,9a 400,1 60,80a P3 (9%) 653,2a 406,2 62,19a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) Tabel 3. Rataan bobot potong dan bobot karkas ayam broiler umur 35 hari Berdasarkan hasil diatas menunjukkan bahwa pemberian tepung singkong fermentasi pada ayam broiler tidak menunjukkan perberbedaan yang nyata (P>0,05) terhadap bobot potong dan bobot karkas. Hal ini sesuai dengan Nahashon et al. (2005) yang menyatakan bahwa bobot karkas ayam broiler berkaitan dengan bobot potong dan pertambahan bobot badan. Semakin tinggi bobot potong broiler, semakin tinggi bobot karkas yang dihasilkan, begitupun sebaliknya. Hal ini diduga karena tepung singkong fermentasi lebih terkonsentrasi dalam meningkatkan bobot non karkas. Ransum yang diberikan dengan tepung singkong fermentasi hingga 9% dari jumlah ransum menghasilkan presentase bobot potong dari masing-masing perlakuan hampir seragam. Hal ini juga menunjukkan bahwa meskipun hasilnya belum memperlihatkan perbedaan yang nyata secara statistik, namun pemberian tepung singkong tanpa fermentasi lebih baik dalam meningkatkan bobot karkas dibandingkan jika difermentasi. Bobot karkas merupakan gambaran dari pertumbuhan jaringan dan tulang. Tingginya bobot karkas dipengaruhi oleh bobot hidup dan komponen non karkas. Hal ini sejalan dengan Panjaitan et al. (2012) yang menyatakan bahwa faktor – faktor yang dapat mempengaruhi bobot karkas diantaranya yaitu umur, kandungan nutrisi pakan, laju pertumbuhan, konformasi tubuh dan bobot tubuh ternak serta jenis ternak yang digunakan. y g g Pemberian tepung singkong tanpa fermentasi sangat baik untuk meningkatkan bobot karkasnya, meskipun penggunaan tepung singkong fermentasi sebesar 9% tidak memperlihatkan penurunan presentase karkas ayam broiler yang rendah. Hal ini dikarenakan hasil fermentasi tepung singkong mengakibatkan peningkatan kandungan protein, sehingga terjadi peningkatan kandungan asam amino yang dibutuhkan untuk peningkatan karkas ayam pedaging. Nutrien yang berkaitan dengan proses pembentukan karkas adalah protein kasar. Protein merupakan penyusun jaringan sel tubuh dan kandungan protein dalam ransum sangat mempengaruhi pencapaian bobot badan ternak (Jumiati dkk., 2017). Bobot Hati Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap bobot hati ayam broiler umur 35 hari dapat dilihat pada Tabel 4. Tabel 4. Rataan Bobot Hati Ayam Broiler Perlakuan Bobot Hati (g) Persentase Hati (%) P0 16,5 2,466a P1 14,8 2,402 a P2 17,3 2,606 a P3 18,3 2,812 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) 399 p-ISSN 1412-1468 e-ISSN 2355-3545 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 Hasil analisis penelitian persentase hati ayam terhadap pemberian tepung singkong fermentasi dan tepung singkong tanpa fermentasi tidak menunjukkan perbedaan yang nyata (P>0,05). Rata-rata bobot hati ayam broiler berkisar antara 14,8-18,3 g (2,402-2,81% dari bobot potong). Menurut Yuwanta (2004), berat hati adalah 3% dari bobot badan ayam broiler. Meskipun berpengaruh tidak nyata (P>0,05), namun pemberian tepung singkong yang difermentasi pada level pemberian 9% kedalam ransum terlihat meningkatkan bobot hati. Meningkatnya bobot hati disebabkan karena semakin tinggi level tepung singkong yang diberikan, HCN semakin meningkat, Hasil analisis penelitian persentase hati ayam terhadap pemberian tepung singkong fermentasi dan tepung singkong tanpa fermentasi tidak menunjukkan perbedaan yang nyata (P>0,05). Rata-rata bobot hati ayam broiler berkisar antara 14,8-18,3 g (2,402-2,81% dari bobot potong). Menurut Yuwanta (2004), berat hati adalah 3% dari bobot badan ayam broiler. Meskipun berpengaruh tidak nyata (P>0,05), namun pemberian tepung singkong yang difermentasi pada level pemberian 9% kedalam ransum terlihat meningkatkan bobot hati. Meningkatnya bobot hati disebabkan karena semakin tinggi level tepung singkong yang diberikan, HCN semakin meningkat, sehingga hati bekerja ekstra untuk meningkatkan produksi dan sekresi empedu guna menetralisir HCN tersebut, sebagai konsekuensinya ukuran hati meningkat. Pernyataan ini sesuai dengan pendapat Sinurat (2002), yang mengemukakan peningkatan berat hati disebabkan oleh penyakit atau racun yang terbawa bersama makanan. Proventikulus Ayam Broiler 400 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 Ventrikulus Ayam Broiler Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap bobot ventrikulus ayam broiler dapat dilihat pada Tabel 6. Tabel 6. Rataan Bobot Ventrikulus Ayam Broiler Setelah Dibersihkan Perlakuan Rataan Bobot Ventrikulus Setelah Dibersihkan (g) Persentase Ventrikulus (%) P0 12,4 1,866 a P1 10,7 1,644 a P2 10,9 1,646 a P3 11,7 1,784 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) Ventrikulus Ayam Broiler Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap bobot ventrikulus ayam broiler dapat dilihat pada Tabel 6. terhadap bobot ventrikulus ayam broiler dapat dilihat pada Tabel 6. Ventrikulus Ayam Broiler Tabel 6. Rataan Bobot Ventrikulus Ayam Broiler Setelah Dibersihkan Perlakuan Rataan Bobot Ventrikulus Setelah Dibersihkan (g) Persentase Ventrikulus (%) P0 12,4 1,866 a P1 10,7 1,644 a P2 10,9 1,646 a P3 11,7 1,784 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) Tabel 6. Rataan Bobot Ventrikulus Ayam Broiler Setelah Dibersihkan Tabel 6 menujukkan bahwa rataan persentase bobot ventrikulus ayam broiler pada setiap perlakuan tidak berbeda nyata. Persentase bobot ventrikulus pada penelitian ini berkisar antara 1,64-1,86%. Ventrikulus berperan dalam mencerna ransum secara mekanik. Berdasarkan penelitian Mahmilia (2005) bahwa perkembangan organ ventrikulus dipengaruhi oleh aktivitas kerja ventrikulus dalam mencerna serat kasar pakan. Hasil penelitian ini sesuai dengan kandungan persentase ventrikulus unggas yang normal yaitu 1,6-2,3% dari bobot potong (Simamora, 2011). Persentase ventrikulus paling tinggi pada perlakuan P0, meskipun belum menunjukkan perbedaan yang nyata dengan perlakuan lainnya. Tingginya persentase bobot ventrikulus pada P0 dapat disebabkan karena serat kasar yang tinggi dalam tepung singkong tanpa fermentasi, sehingga mengakibatkan kerja ventrikulus lebih berat, sesuai dengan pendapat Ciptaan dan Mirnawati (2001), yang mengemukakan bahwa berat ventrikulus dipengaruhi serat kasar ransum. Semakin tinggi kadar serat kasar ransum maka ventrikulus akan lebih berat. Selain itu serat yang tinggi dalam pakan akan memperbesar ukuran ventrikulus, karena organ tersebut dipacu untuk lebih banyak bekerja secara fisiologis dalam memproses pencernaan serat, baik secara mekanik maupun enzimatis. Mahmilia (2005) juga menyatakan bahwa otot dinding organ ventrikulus akan bekerja lebih keras apabila kadar serat kasar yang dicerna semakin tinggi, sehingga mampu meningkatkan perkembangan organ ventrikulus. Berdasarkan penelitian Anggoro et al. (2019) menyatakan bahwa aktivitas kerja pencernaan organ ventrikulus yang meningkat akan mengakibatkan perubahan terhadap morfologi saluran tersebut sehingga dapat berkembang lebih optimal. Proventikulus Ayam Broiler Hasil pengamatan pengaruh pemberian tepung singkong yang fermentasi terhadap bobot proventikulus ayam broiler dapat dilihat pada Tabel 5. Tabel. 5. Rataan Bobot Proventikulus Ayam Broiler Perlakuan Rataan Bobot Proventikulus Setelah Dibersihkan (g) Persentase Proventikulus (%) P0 3,9 0,582a P1 3,2 0,47 a P2 3,1 0,468 a P3 3,5 0,53 a Keterangan : Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) Tabel. 5. Rataan Bobot Proventikulus Ayam Broiler Hasil analisis sidik ragam bobot proventikulus ayam broiler menunjukkan bahwa pemberian tepung singkong fermentasi tidak menunjukkan hasil yang berbeda nyata pada setiap perlakuan. Peran dari proventrikulus yaitu sebagai tempat terjadinya proses pencernaan protein pakan secara kimiawi atau enzimatis. Berdasarkan penelitian Handoko et al. (2013) bahwa dalam mencerna protein ransum, pepsinogen akan disekresi oleh organ proventrikulus. Proventrikulus berfungsi mencerna protein yang dibantu oleh enzim pepsin, proses tersebut menghasilkan hydrochloric acid (HCI) yang disekresikan oleh grandular cell. Hasil persentase bobot proventikulus pada setiap perlakuan belum mendekati 1% dari bobot potong, namun masih berada dalam kisaran normal dan sesuai dengan pendapat Elfiandra (2007). Siregar (2011) juga menyatakan bahwa rataan persentase bobot proventrikulus berkisar 0,45%-0,56%. Hasil penelitian ini membuktikan bahwa perlakuan pakan, tidak mempengaruhi fungsi dan kinerja dari proventrikulus. Hal ini sesuai dengan penelitian Iskandar (2004) yang menyatakan bahwa faktor yang mempengaruhi perkembangan organ proventrikulus salah satunya yaitu aktivitas pencernaan protein ransum. Rataan persentase bobot proventrikulus ayam broiler pada setiap perlakuan tidak berbeda nyata, hal ini dikarenakan kandungan protein pakan yang diberikan pada semua perlakuan sama. Kondisi ini menyebabkan intake protein relatif sama, sehingga kerja proventrikulus dalam mensekresikan pepsin untuk pencernaan protein tidak berbeda. Berdasarkan penelitian Sugiharto et al. (2018) bahwa apabila aktivitas pencernaan dalam organ proventrikulus meningkat akan mengakibatkan perubahan terhadap morfologi saluran tersebut sehingga mampu berkembang lebih optimal. Hasil penelitian ini membuktikan bahwa perlakuan pakan, tidak mempengaruhi fungsi dan kinerja dari proventrikulus. Hal ini sesuai dengan penelitian Iskandar (2004) yang menyatakan bahwa faktor yang mempengaruhi perkembangan organ proventrikulus salah satunya yaitu aktivitas pencernaan protein ransum. Rataan persentase bobot proventrikulus ayam broiler pada setiap perlakuan tidak berbeda nyata, hal ini dikarenakan kandungan protein pakan yang diberikan pada semua perlakuan sama. Kondisi ini menyebabkan intake protein relatif sama, sehingga kerja proventrikulus dalam mensekresikan pepsin untuk pencernaan protein tidak berbeda. Berdasarkan penelitian Sugiharto et al. (2018) bahwa apabila aktivitas pencernaan dalam organ proventrikulus meningkat akan mengakibatkan perubahan terhadap morfologi saluran tersebut sehingga mampu berkembang lebih optimal. Proventikulus Ayam Broiler Hal ini mengakibatkan beban ventrikulus menjadi lebih besar untuk memperkecil ukuran partikel ransum secara fisik, sehingga urat daging ventrikulus akan menjadi lebih tebal. Hal ini juga akan berdampak pada membesarnya ukuran ventrikulus, meskipun hal tersebut tidak mempengaruhi fungsi fisiologis dari ventrikulus (Rosyani, 2013). Tabel 7. Rataan Bobot Usus Halus Ayam Broiler y Perlakuan Rataan Berat Usus Halus (g) Persentase Berat Usus Halus (%) P0 6,8 1,03a P1 6 0,894 a P2 7,1 1,078 a P3 6,6 1,008 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) y Perlakuan Rataan Berat Usus Halus (g) Persentase Berat Usus Halus (%) P0 6,8 1,03a P1 6 0,894 a P2 7,1 1,078 a P3 6,6 1,008 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) 401 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 Bobot dan Panjang Usus Halus persentase normal berat usus halus yaitu 2,31-2,49%. Perkembangan usus halus unggas sangat dipengaruhi oleh kandungan serat kasar dalam ransum yang dikonsumsi oleh ayam broiler. Serat kasar merupakan salah satu zat makanan penting dalam ransum unggas, karena berfungsi merangsang gerak peristaltik saluran pencernaan sehingga proses pencernaan zat-zat makanan berjalan dengan baik. Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap bobot usus halus ayam broiler dapat dilihat pada Tabel 7. Hasil penelitian pada setiap perlakuan memperoleh persentase bobot usus halus berkisar antara 0,89-1,07%. Hasil analisis ragam pada penelitian ini menunjukkan bahwa tidak ada perbedaan yang nyata terhadap persentase bobot usus halus akibat pemberian tepung singkong fermentasi. Persentase bobot usus halus hasil penelitian ini lebih rendah dari pendapat Elfiandra (2007) yang mengemukakan bahwa Tabel 8. Rataan panjang usus halus ayam broiler Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap panjang usus halus ayam broiler dapat dilihat pada Tabel 8. Tabel 8. Rataan panjang usus halus ayam broiler Perlakuan Rataan Panjang Usus Halus (cm) P0 24,07 P1 23,57 P2 24,70 P3 23,26 Keterangan: Tidak berbeda nyata (P>0,05) singkong tanpa fermentasi sehingga peningkatan kadar serat kasar dalam ransum cenderung memperpanjang usus halus. Semakin tinggi serat kasar dalam ransum, maka semakin lambat laju pencernaan dan penyerapan zat makanan. Berdasarkan Tabel 8. terlihat bahwa panjang usus halus ayam broiler berkisar 23,36-24,7cm. Hasil ini menunjukkan bahwa panjang usus halus hasil penelitian ini lebih pendek dibandingkan dengan hasil penelitian Suprijatna et al., (2008), yang mencapai 150cm. Panjang usus pemberian tepung singkong tanpa fermentasi lebih panjang dibandingkan dengan hasil pemberian tepung singkong fermentasi. Hal ini diduga karena tingginya kandungan serat kasar pada tepung Bobot dan Panjang Usus Besar Hasil pengamatan pengaruh pemberian tepung singkong fermentasi terhadap berat usus besar ayam broiler dapat dilihat pada Tabel 9 Tabel 9. Rataan Bobot Usus Besar Ayam Broiler Perlakuan Rataan Bobot Usus Besar (g) Persentase Bobot Usus Besar (%) P0 1,8 0,27a P1 1,9 0,27 a P2 2,2 0,33 a P3 2,1 0,32 a Keterangan: Nilai rataan yang diikuti huruf yang sama ke arah kolom menunjukkan tidak berbeda nyata (P>0,05) Tabel 9. Rataan Bobot Usus Besar Ayam Broiler adalah untuk perombakan partikel pakan yang tidak tercerna oleh mikroorganisme menjadi feses. Berdasarkan hasil penelitian ini persentase usus besar ayam broiler berkisar antara 0,27-0,332%. Hal ini menunjukkan bahwa persentase bobot usus besar diatas rata-rata ukuran usus besar yang normal yaitu 0.17-0.20% (Neil, 2008). Fungsi usus besar Hasil pengamatan pengaruh pemberian tepung singkong fermentasi 402 Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 p-ISSN 1412-1468 e-ISSN 2355-3545 ZIRAA’AH, Volume 48 Nomor 3, Oktober 2023 Halaman 394-404 terhadap panjang usus besar ayam broiler dapat dilihat pada Tabel 10. Tabel 10. Rataan Panjang Usus Besar Ayam Broiler Perlakuan Rataan Panjang Usus Besar (cm) P0 6,36 P1 6,42 P2 6,88 P3 6,64 Keterangan: Tidak berbeda nyata (P>0,05) terhadap panjang usus besar ayam broiler dapat dilihat pada Tabel 10. Keterangan: Tidak berbeda nyata (P>0,05) Cassava Peel. Academic Journals 4(3): 51- 56. Berdasarkan hasil penelitian ini persentase panjang usus besar ayam broiler berkisar antara 0,94-1,04%. Menurut Ibrahim (2008), ukuran panjang usus akan bertambah seiring dengan laju pertumbuhan dan konsumsi pakan ayam broiler. Serat kasar dalam ransum akan mempengaruhi panjang usus besar ayam broiler. Hal ini sesuai dengan pernyataan Amrullah (2004), bahwa ukuran panjang, tebal, bobot berbagai saluran pencernaan dapat berubah selama perkembangan dan dipengaruhi oleh ransum. Ransum dengan kandungan serat kasar yang tinggi dapat memperberat, memperpanjang dan mempertebal berbagai saluran pencernaan. Amri, E. dan Pratiwi P. 2014. Pembuatan Mocaf (Modified Cassafa Flour) dengan Proses Fermentasi Menggunakan Berbagai Jenis Ragi. Jurnal Pelangi (2):171-179. Amrullah, I. K. 2004. Nutrisi Ayam Broiler. Cetakan ke-2. Lembaga Satu Gunung Budi. Bogor. Anggoro, L., H. I. Wahyuni, dan E. Widiastuti. 2019. Pengaruh Pemberian Kulit Pisang Fermentasi Dengan Chrysonilia Crassa dan Bacillus Subtilis Terhadap Bobot Relatif Organ Pencernaan Ayam Broiler. J. Peternakan. 11(4): 1-7. KESIMPULAN Berdasarkan hasil penelitian, maka dapat disimpulkan bahwa pemberian tepung singkong tanpa fermentasi mampu meningkatkan bobot karkas ayam broiler dibandingkan dengan tepung singkong fermentasi. Hasil evaluasi terhadap organ pencernaan diketahui bahwa pemberian tepung singkong fermentasi sebesar 6% mampu meningkatkan bobot ventrikulus, usus halus dan usus besar, sedangkan pemberian tepung singkong sebesar 9% mampu meningkatkan bobot hati. Badan Pusat Statistik. 2015. Produksi Singkong di Kota Palangka Raya Kalimantan Tengah. 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Perubahan Nilai Gizi Tepung Eceng Gondok Fermentasi dan Pemanfaatannya Sebagai Ransum Ayam Pedaging. JITV. 10 (2) 90–95. Mahmilia, F. 2005. Perubahan Nilai Gizi Tepung Eceng Gondok Fermentasi dan Pemanfaatannya Sebagai Ransum Ayam Pedaging. JITV. 10 (2) 90–95. Hardini, S.Y.P.K. 2003. Peningkatan Bobot Badan Ayam Merawang yang dipelihara Bersama Ayam Broiler dengan Memperhatikan Perilaku Makanannya. Skripsi. FMIPA Universitas Terbuka. Nahashon, S. N., Adefope, N., Amenyenu, A., & Wright, D. 2005. Effects of Dietary Metabolizable Energy and Crude Protein Concentration on Growth Performance and Carcass Characteristics of French Guinea Broiler. Poultry Science, 84, 337– 344. Ibrahim, S. 2008. Hubungan Ukuran-ukuran Usus Halus dengan Berat Badan Broiler. J. Agribisnis Peternakan. 8 (2): 42-46. Neil, A. C. 2008. Biology 2nd edition.The Benjamin Coming Publishing Company Inc., Pec Wood City. Iskandar, S. 2004. 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https://openalex.org/W2130728804	https://europepmc.org/articles/pmc1420271?pdf=render	English	null	Identification and targeted disruption of the mouse gene encoding ESG1 (PH34/ECAT2/DPPA5)	BMC developmental biology	2,006	cc-by	6,272	Published: 28 February 2006 Published: 28 February 2006 BMC Developmental Biology2006, 6:11 doi:10.1186/1471-213X-6-11 This article is available from: http://www.biomedcentral.com/1471-213X/6/11 © 2006Amano et al; licensee BioMed Central Ltd. ; This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. BioMed Central BioMed Central Abstract Background: Embryonic stem cell-specific gene (ESG) 1, which encodes a KH-domain containing protein, is specifically expressed in early embryos, germ cells, and embryonic stem (ES) cells. Previous studies identified genomic clones containing the mouse ESG1 gene and five pseudogenes. However, their chromosomal localizations or physiological functions have not been determined. Results: A Blast search of mouse genomic databases failed to locate the ESG1 gene. We identified several bacterial artificial clones containing the mouse ESG1 gene and an additional ESG1-like sequence with a similar gene structure from chromosome 9. The ESG1-like sequence contained a multiple critical mutations, indicating that it was a duplicated pseudogene. The 5' flanking region of the ESG1 gene, but not that of the pseudogene, exhibited strong enhancer and promoter activity in undifferentiated ES cells by luciferase reporter assay. To study the physiological functions of the ESG1 gene, we replaced this sequence in ES cells with a β-geo cassette by homologous recombination. Despite specific expression in early embryos and germ cells, ESG1-/- mice developed normally and were fertile. We also generated ESG1-/- ES cells both by a second independent homologous recombination and directly from blastocysts derived from heterozygous intercrosses. Northern blot and western blot analyses confirmed the absence of ESG1 in these cells. These ES cells demonstrated normal morphology, proliferation, and differentiation. Conclusion: The mouse ESG1 gene, together with a duplicated pseudogene, is located on chromosome 9. Despite its specific expression in pluripotent cells and germ cells, ESG1 is dispensable for self-renewal of ES cells and establishment of germcells. tain pluripotency, which is the ability to differentiate into a wide variety of cells, and rapid proliferation. These char- acteristics make mouse ES cells an essential component of gene targeting technology. These qualitiesalso make BMC Developmental Biology Open Access Hisayuki Amano1,3, Ken Itakura1, Masayoshi Maruyama1, Tomoko Ichisaka1,2, Masato Nakagawa1 and Shinya Yamanaka1,2 Address: 1Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan, 2CREST, Japan Science and Technology Cooperation, Kawaguchi, Japan and 3Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita, Japan Email: Hisayuki Amano - h-amano@molbio.med.osaka-u.ac.jp; Ken Itakura - k-itakura@kose.co.jp; Masayoshi Maruyama - maruyama@frontier.kyoto-u.ac.jp; Tomoko Ichisaka - ichisaka@frontier.kyoto-u.ac.jp; Masato Nakagawa - nakagawa@frontier.kyoto-u.ac.jp; Shinya Yamanaka - yamanaka@frontier.kyoto-u.ac.jp * Corresponding author Received: 29 December 2005 Accepted: 28 February 2006 http://www.biomedcentral.com/1471-213X/6/11 http://www.biomedcentral.com/1471-213X/6/11 http://www.biomedcentral.com/1471-213X/6/11 BMC Developmental Biology 2006, 6:11 human ES cells attractive sources for cell transplantation therapy to treat various diseases, including spinal cord injuries and juvenile diabetes. The molecular mechanisms underlying the pluripotency and rapid proliferation of ES cells are currently a major focus of the field of stem cell biology [4-6]. noma cells [13]. The expression of this gene was confined in mice to early embryos and germ cells [14]. It is also expressed in pluripotent cells, including ES cells, embry- onic germ cells, and multipotent germline stem cells [15]. ESG1 encodes a polypeptide of 118 amino acids that con- tains a single KH domain, which is an RNA-binding domain [16]. It remains unclear, however, if ESG1 func- tions as an RNA-binding protein or the roles it plays in ES cells and mice. To identify molecules essential in ES cells for these prop- erties, several groups have utilized transcriptome analyses to identify genes specifically expressed in ES cells and early embryos. These analyses, including DNA microarray analyses [7] and expressed sequence tag analyses [8-12], identified several common transcripts, including ESG1 that was also designated dppa5 or ECAT2. Previous genomic library screening by identified genomic clones containing the mouse ESG1 gene and seven pseu- dogenes [17]. Two of these pseudogenes exhibit a similar exon-intron structure as the ESG1 gene, indicating their generation by gene duplication. The five remaining pseu- dogenes did not contain any introns, indicating that these were generated by retrotransposition of ESG1 transcripts. ESG1 was originally identified as a transcript Ph34 that was down-regulated by retinoic acid in embryonic carci- ESG1 pseudogenes identified by a Blast search of mouse genomic databases Figure 1 ESG1pseudogenes identified by a Blast search of mouse genomic databases. Substitution mutations are indicated by black lines. Intron-like gap sequences are indicated with triangles. Chromosomal localizations are shown on the right. chromosome 5 11 12 14 16 17 X unknown unknown 9 1 NNNNNNN ESG1 pseudogenes identified by a Blast search of mouse genomic databases Figure 1 ESG1pseudogenes identified by a Blast search of mouse genomic databases. Substitution mutations are indicated by black lines. Intron-like gap sequences are indicated with triangles. Chromosomal localizations are shown on the right. chromosome 5 11 12 14 16 17 X unknown unknown 9 1 NNNNNNN ESG1 pseudogenes identified by a Blast search of mouse genomic databases Figure 1 ESG1pseudogenes identified by a Blast search of mouse genomic databases. Background g Embryonic stem (ES) cells were first derived from the blas- tocysts of mice in 1981 [1,2] and humans in 1998 [3]. ES cells have two important properties: theability to main- Page 1 of 9 (page number not for citation purposes) Page 1 of 9 (page number not for citation purposes) http://www.biomedcentral.com/1471-213X/6/11 Targeted disruption of the mouse ESG1 gene Targeted disruption of the mouse ESG1 gene To study the function of ESG1, we deleted the gene by homologous recombination in mouse ES cells. We replaced the three exons with either a fusion of the neo- mycin-resistance and β-galactosidase genes (β-geo) or the hygromycin resistant gene (HygR) using two targeting vec- tors (Figure 4A) introduced into RF8 ES cells by electropo- ration. We obtained eight ES cell clones with correct homologous recombination of the β-geo targeting vector, which was confirmed by Southern blot analysis (Figure 4B). We obtained only one clone with correct homolo- gous recombination of the HygR targeting vector. On chromosome 9, we identified a DNA fragment similar to the ESG1 gene that included two putative introns. These putative first and second exons, however, contained (4) multiple mutations of the ESG1 cDNA sequence. The putative third exon was identical to that of the previously reported ESG1 gene. Also on chromosome 9, we identi- fied another DNA fragment that was similar, but not iden- tical, to the third exon of the ESG1 gene. These findings suggest that these ESG1-like sequences on chromosome 9 have not been correctly assembled. To obtain homozygous mutant ES cells, we introduced the β-geo vector into HygR heterozygous ES cells. Of 105 G418-resistant colonies tested, 49 were homozygous for ESG1 deletion. Northern blot and western blot analyses confirmed the absence of ESG1 in these cells (Figure 4C). In 29 clones, the β-geo vector had replaced the HygR vec- tor, such that the cells remained heterozygous. In the remaining 27 clones, the β-geo vector was integrated at non-homologous sites. To obtain DNA fragments containing the ESG1 gene, we screened the bacterial artificial chromosome (BAC) DNA pool by PCR using primer pairs that would only amplify the real gene, not the pseudogenes. We obtained two inde- pendent, but overlapping BAC clones. Southern blot anal- yses and sequencing demonstrated that these clones contained a sequence exhibiting complete identity with ESG1 cDNA that was interrupted by two putative introns (Figure 2A). The two intron sequences begin with GT and terminate with AG, fulfilling the GT-AG rule of exon- intron junctions [19]. The 5' flanking region of this DNA fragment exhibited strong promoter/enhancer activity by luciferase reporter assays in undifferentiated ES cells, but not in somatic cells (Figure 3). The same fragment showed much weaker activity after induction of differentiation by retinoic acid. http://www.biomedcentral.com/1471-213X/6/11 BMC Developmental Biology 2006, 6:11 The chromosomal localizations of the mouse ESG1 gene and pseudogenes, however, have not been reported. The chromosomal localizations of the mouse ESG1 gene and pseudogenes, however, have not been reported. We also found that the two BAC clones contained another ESG1-like sequence (Figure 2A). The two sequences, sepa- rated by a 68 kbp intergenic sequence, were oriented in opposite directions. The ESG1-like sequence exhibited greater than 95% identity to the exons and introns of the ESG1 gene. This sequence, however, contained critical nucleotide substitutions in all of the exons and one nucle- otide insertion in exon 2 (Figure 2B). Although 675 base pairs of the 3' flanking regions were conserved between the ESG1 gene and the pseudogene, only five base pairs of the 5' flanking region were identical. This 5' flanking region (~6 kbp) did not possess any promoter/enhancer activity in luciferase reporter assays (Figure 3). It is thus unlikely that this sequence is transcribed or translated into a functional protein. This sequence likely represents a duplication pseudogene. Bierbaum previously reported the existence of two pseudogenes with similar exon-intron organization as the ESG1 gene [17]. We could not deter- mine which of these two pseudogenes corresponds to the one we identified or the location of the remaining pseu- dogene. In this study, we determined the structure of the mouse gene encoding this protein and related pseudogenes. We also performed gene targeting to determine the physiolog- ical function of ESG1. Chromosomal localization and structures of mouse ESG1 gene and psedogenes To determine the chromosomal localizations of the mouse ESG1 gene and pseudogenes, we performed a Blast analysis of the mouse genomic database with the ESG1 cDNA sequence as a query. We identified several putative pseudogenes without introns on chromosomes 1, 5, 11, 12, 14, 16, 17, and X (Figure 1). In addition, two intron- less pseudogenes were identified in DNA fragments for which the chromosomal localization remained unmapped. While these pseudodgenes have significant homology to ESG1 cDNA, they could not produce func- tional proteins, because of critical mutations. This result suggests that there are a larger number of intron-less pseu- dogenes than previously anticipated. Existence of multi- ple retropseudogenes is a hallmark of pluripotent cell- specific genes [18]. http://www.biomedcentral.com/1471-213X/6/11 Substitution mutations are indicated by black lines. Intron-like gap sequences are indicated with triangles. Chromosomal localizations are shown on the right. http://www.biomedcentral.com/1471-213X/6/11 http://www.biomedcentral.com/1471-213X/6/11 Page 3 of 9 (page number not for citation purposes) Targeted disruption of the mouse ESG1 gene Histo- logical examination of testis and ovary could not identify any abnormalities (not shown). These data demonstrated that ESG1 is dispensable for both mouse development and germ cell formation. We examined the gene expression profiles of ESG1-/- ES cells using oligonucleotide-based DNA microarrays repre- senting ~20,000 genes. In comparison to control ES cells, ESG1 was identified as the gene reduced to the greatest extent in ESG1-/- ES cells (Figure 6A). The expression of ES cell marker genes, such as Nanog and Oct3/4, was normal in ESG1-/- ES cells. We confirmed normal Oct3/4 expres- sion at protein levels by Western blot (Figure 6B). The overall gene expression profiles were similar between con- trol ES cells and ESG1-/- ES cells. Several genes exhibited a greater than two-fold reduction in ESG1-/- cells, including Krt1-8, Pem, Ctgf, Ptgs2, Igf2 and Inhba. These genes might be regulated directly or indirectly by ESG1. Since ESG1 contains a KH-type RNA-binding domain, it may stabilize mRNA of these genes. Further studied are required to clar- ify this possibility. We also generated ES cells from blastocysts obtained by intercrosses of ESG1+/- males and ESG1-/- females. Of the eight ES cell lines established, two clones were ESG1-/-. These ESG1-null ES cells demonstrated normal morphol- ogy, proliferation, and differentiation (not shown), con- firming that ESG1 is dispensable in ES cells. Targeted disruption of the mouse ESG1 gene We concluded that this sequence is the ESG1 gene. ESG1-/- ES cells exhibited normal morphology (Figure 5A). These cells also proliferated at a speed comparable to that of the control (heterozygous and wild-type) cells (Figure 5B). ESG1-/- cells differentiated normally after the removal of leukemia inhibitory factor (Figure 5B) or retin- oic acid treatment (not shown). When transplanted into hind flanks of nude mice, these cells produced teratomas, tumors containing components of all three germ layers (Figure 5C). These results indicate that ESG1 is dispensa- Page 3 of 9 (page number not for citation purposes) BMC Developmental Biology 2006, 6:11 http://www.biomedcentral.com/1471-213X/6/11 BAC clones containing the ESG1 gene and a duplication pseudogene (PS) Figure 2 BAC clones containing the ESG1 gene and a duplication pseudogene (PS). A) Localization of the gene and PS on chromosome 9. B) Sequence comparison of the gene and PS revealed that these are homologous from eight base pairs upstream of the first exon to 675 bp downstream of the third exon. Substitution mutations are indicated by black lines. The insertion is indicated by an open triangle. ESG1 exon PS 68kbp ESG1 PS 675bp 1 2 3 8bp A. B. 2 1 3 2 1 3 A. B. BAC clones containing the ESG1 gene and a duplication pseudogene (PS) Figure 2 BAC clones containing the ESG1 gene and a duplication pseudogene (PS). A) Localization of the gene and PS on chromosome 9. B) Sequence comparison of the gene and PS revealed that these are homologous from eight base pairs upstream of the first exon to 675 bp downstream of the third exon. Substitution mutations are indicated by black lines. The insertion is indicated by an open triangle. ble for the self-renewal properties and pluripotency of ES cells. We obtained germline transmission from three clones. We obtained ESG1-/- mice at the Mendelian ratios (36 wild- type, 69 ESG1+/-, and 45 ESG1-/-) from intercrosses of ESG1+/- mice. These animals exhibited normal develop- ment, gross appearance, and fertility (not shown). Histo- logical examination of testis and ovary could not identify any abnormalities (not shown). These data demonstrated that ESG1 is dispensable for both mouse development and germ cell formation. We obtained germline transmission from three clones. We obtained ESG1-/- mice at the Mendelian ratios (36 wild- type, 69 ESG1+/-, and 45 ESG1-/-) from intercrosses of ESG1+/- mice. These animals exhibited normal develop- ment, gross appearance, and fertility (not shown). Identification and analyses of BAC clones containing the mouse ESG1 gene g To identify bacterial artificial chromosome (BAC) clones containing mouse ESG1 gene, we performed PCR-based screening of mouse BAC library DNA pools (Research Genetics) using the pH34-u38 (5'-GAAGTCTGGTTCCTT- GGCAGG-3') and pH34-L394 (5'-ACTCGATACACT- GGCCTAGC-3') primers. Following restriction enzyme digestion, we performed Southern blot analyses of BAC clones as described [20] using the pH34-U258 (5'-CTC- GAGTGTACAGTCAAGTGGTTGCTGGGA-3'), pH34-U65 (5'-GTGACCCTCGTGACCCGTAA-3'), pH34-intron1L (5'-CTGCGTGAGAGAAACACCAAACAGGC-3'), pH34- L545 (5'-TGTGAATGGGAAGGTTACCACTCT-3') and pH34-SCL1 (5'-GCCCTCTTCTGGTTTGTCTCGAAAT-3') probes. Hybridization with these probes revealed bands containing either the ESG1 gene or pseudogenes. Conclusion To analyze the physiological roles of ESG1, we identified the mouse gene on chromosome 9 and deleted it by homologous recombination in ES cells. Despite specific expression in early embryos, germ cells, and pluripotent To generate ESG1-knockout mice, we injected β-geo- ESG1+/- ES cell clones into the blastocysts of C57BL6 mice. Page 4 of 9 (page number not for citation purposes) BMC Developmental Biology 2006, 6:11 http://www.biomedcentral.com/1471-213X/6/11 Promoter/enhancer activity of the ESG1 gene and pseudogene Figure 3 Promoter/enhancer activity of the ESG1 gene and pseudogene. DNA fragments of ~6 kbp isolated from the 5' flanking regions of the gene and PS were transferred into luciferase reporter plasmids. We introduced the reporter genes into undiffer- entiated ES cells (open columns), retinoic acid-treated ES cells (grey columns), and NIH3T3 cells (closed columns). Data repre- sent the averages and standard deviations of three independent experiments. PS 0 100 200 -6kbp -6kbp Luc ESG1 LUC activity (RLU) 300 PS -6kbp -6kbp ESG1 0 y g p g g Promoter/enhancer activity of the ESG1 gene and pseudogene. DNA fragments of ~6 kbp isolated from the 5' flanking regions of the gene and PS were transferred into luciferase reporter plasmids. We introduced the reporter genes into undiffer- entiated ES cells (open columns), retinoic acid-treated ES cells (grey columns), and NIH3T3 cells (closed columns). Data repre- sent the averages and standard deviations of three independent experiments. cloned into pBluescript KS(-) for sequencing. To sequence the ESG1 pseudogene and the 3' flanking region, an 8 kbp NotI/XhoI fragment was cloned into pBluescript KS(-). BamHI- or PstI- fragments of this vector were also cloned into pBluescript KS(-). To identify the sequence contain- ing the 5' flanking regions of the ESG1 gene and the related pseudogenes, we used a TOPO walker kit (Invitro- gen) with the pH34-T2L (5'-ACTAGTCGCAGCAG- GGATCCAGGAATATCT-3') and pH34-L394 primers. The resulting sequence was cloned into pCR2.1 (Invitrogen). We obtained a ~6 kbp band from the NsiI-digensted library; XbaI-, SpeI-, EcoRI-, and PstI-digested fragments of this band were cloned into pBluescript KS(-) for sequenc- ing. This fragment contained the 5' flanking region of the ESG1 gene. A ~3 kbp fragment, obtained from the SacI- digested library, was cloned into pCR2.1 for sequencing. This fragment was contained the 5' region flanking the pseudogene. cells, our data demonstrated that ESG1 is dispensable for mouse development, germ cell formation, and ES cell self- renewal. Construction of ESG1 targeting vectors We replaced all of the ESG1 exons with two targeting vec- tors containing either an IRES-β-geo cassette [21] or an IRES-HygR cassette by promoter trap selection. We ampli- fied the 5' arm (1.8 kbp) using KOD plus (TOYOBO) with the pH34-targetpair5-U (5'-CCGCGGAAAGTCAAGA- GATTGGGTGG-3') and pH34-targetpair5-L (5'-GCG- GCCGCCTTTACGGGTCACGAGGGTCAC-3') primers. To sequence the region containing the mouse ESG1 gene and the 3' flanking region, we subcloned a ~15 kbp XhoI- SalI fragment into the pZERO-2 vector (Invitrogen). Hin- dIII- or EcoRI-digested fragments of this vector were then Page 5 of 9 (page number not for citation purposes) Page 5 of 9 (page number not for citation purposes) BMC Developmental Biology 2006, 6:11 http://www.biomedcentral.com/1471-213X/6/11 Targeted disruption of the mouse ESG1 gene Figure 4 Targeted disruption of the mouse ESG1 gene. A) Targeting strategy. Homologous regions are indicated by thick lines. Recognition sites of PstI (P) and SpeI (S), which were used for Southern blot analyses, are shown. The gene encoding diphtheria toxin A (DTA) was inserted at the 3' end of the targeting vectors to facilitate negative selection. B) Southern blot analyses con- firming homologous recombination. WT, wild-type ES cells; β, β-geo +/- ES cells; H, HygR +/- ES cells; -/-, ESG1-null ES cells. Numbers indicate clone numbers. C) Northern blot (upper) and western blot (lower) analyses of wild-type ES cells (WT), ESG1-null ES cells (-/-, three clones) and heterozygous ES cells (+/-). Northern blot was performed as previously described [20]. To confirm the loading of equal amounts of RNA, ethidium bromide staining of ribosomal RNA is also shown (middle). 1 2 3 P S S P ESG1 locus targeting vector KH domain IRES ββββ-geo pA S P DTA P IRES HygR pA S DTA WT 27 35 36 -/- b 78 H 4 A. B. WT protein RNA rRNA 27 35 36 -/- +/- C. 18 15 12 kbp 1 2 3 P S S P ESG1 locus targeting vector KH domain IRES ββββ-geo pA S P DTA P IRES HygR pA S DTA A. A. 1 2 3 P targeting vector IRES ββββ-geo pA S DTA P IRES HygR pA S DTA WT 27 35 36 -/- b 78 H 4 B. WT protein RNA rRNA 27 35 36 -/- +/- C. 18 15 12 kbp DTA DTA WT 27 35 36 -/- b 78 H 4 B. WT protein RNA rRNA 27 35 36 -/- +/- C. Construction of ESG1 targeting vectors 18 15 12 kbp WT 27 35 36 -/- b 78 H 4 B. protein RNA rRNA C. 18 15 12 kbp B. C. Targeted d Figure 4 Targeted disruption of the mouse ESG1 gene Figure 4 Targeted disruption of the mouse ESG1 gene. A) Targeting strategy. Homologous regions are indicated by thick lines. Recognition sites of PstI (P) and SpeI (S), which were used for Southern blot analyses, are shown. The gene encoding diphtheria toxin A (DTA) was inserted at the 3' end of the targeting vectors to facilitate negative selection. B) Southern blot analyses con- firming homologous recombination. WT, wild-type ES cells; β, β-geo +/- ES cells; H, HygR +/- ES cells; -/-, ESG1-null ES cells. Numbers indicate clone numbers. C) Northern blot (upper) and western blot (lower) analyses of wild-type ES cells (WT), ESG1-null ES cells (-/-, three clones) and heterozygous ES cells (+/-). Northern blot was performed as previously described [20]. To confirm the loading of equal amounts of RNA, ethidium bromide staining of ribosomal RNA is also shown (middle). The 3' arm (5.8 kbp) was amplified using the pH34- targetpair3-U (5'-TGTGGCCAGTGTTTGGTTCT- GGCGGG-3') and pH34-targetpair3-L (5'-CTCGAG- GACTCGCCATTCTAGCCAAG-3') primers. The IRES β- geo or IRES HygR cassettes were ligated in between the two PCR fragments. The diphtheria toxin A cassette was placed downstream of the 3' arm. After linearization with SacII, these targeting vectors were electroporated into 2.0 × 107 RF8 ES cells [22] using a Gene pulser (BIORAD). Transfected cells were selected with 250 µg/mL G418 or 100 µg/mL hygromycin B, respectively. Genomic DNA from G418- or hygromycin B-resistant colonies was screened for homologous recombination by Southern blotting. Southern blot screening for homologous recombination ES cells genomic DNA was extracted with PUREGENE™ Cell Lysis Solution (Gentra systems). For 5' Southern blot analysis, genomic DNA was first digested with PstI, then separated on an 0.8% agarose gel and transferred to a nylon membrane as described [20]. A 560 bp 5' probe was amplified using the ESG1S5 (5'- GATGGTGGTGGTGACT- CAGAG -3') and ESG1AS5 as (5'- CCTCCATTGCCTC- TATATCAG -3') primers. The probe specifically labeled an 18 kbp band from the wild-type locus, a 15 kbp band from the β-geo locus, and a 12 kbp band from the HygR locus. Genomic DNA was also digested with SpeI for 3' Southern blot analysis. Targeted d Figure 4 A 1,010 bp 3' probe was amplified with the pH34U-8000 (5'- CCAACCAGCCAGAGTT- Page 6 of 9 (page number not for citation purposes) Page 6 of 9 (page number not for citation purposes) http://www.biomedcentral.com/1471-213X/6/11 BMC Developmental Biology 2006, 6:11 Analyses of ESG1-null ES cells Figure 5 Analyses of ESG1-null ES cells. A) The morphology of ESG1-null ES cell colonies grown on STO feeder cells. B) Growth curve of wild-type (WT), ESG1-null (-/-) and heterozygous (+/-) ES cells. Each clone (1 × 104 cells/well) was plated in 24-well plates. Cell numbers were determined with a Coulter counter at 2, 4, and 6 days. Data of +/- and -/- cells are shown as aver- ages and standard deviations of three independent clones. C) A section of teratoma derived from ESG1-null ES cells (hematox- ylin & eosin staining). A. 0 5 10 15 20 LIF+ LIF- Cell number ( x105) B. C. +/- -/- WT +/- -/- WT A. 0 5 10 15 20 LIF+ LIF- Cell number ( x105) B. +/- -/- WT +/- -/- WT 0 5 10 15 20 LIF+ LIF- Cell number ( x105) B. +/- -/- WT +/- -/- WT B. A. 0 Analyses of ESG1-null ES cells Figure 5 Analyses of ESG1-null ES cells. A) The morphology of ESG1-null ES cell colonies grown on STO feeder cells. B) Growth curve of wild-type (WT), ESG1-null (-/-) and heterozygous (+/-) ES cells. Each clone (1 × 104 cells/well) was plated in 24-well C. C. Analyses o Figure 5 a yses o SG u S ce s gu e 5 Analyses of ESG1-null ES cells. A) The morphology of ESG1-null ES cell colonies grown on STO feeder cells. B) Growth curve of wild-type (WT), ESG1-null (-/-) and heterozygous (+/-) ES cells. Each clone (1 × 104 cells/well) was plated in 24-well plates. Cell numbers were determined with a Coulter counter at 2, 4, and 6 days. Data of +/- and -/- cells are shown as aver- ages and standard deviations of three independent clones. C) A section of teratoma derived from ESG1-null ES cells (hematox- ylin & eosin staining). y g Analyses of ESG1-null ES cells. A) The morphology of ESG1-null ES cell colonies grown on STO feeder cells. B) Growth curve of wild-type (WT), ESG1-null (-/-) and heterozygous (+/-) ES cells. Each clone (1 × 104 cells/well) was plated in 24-well plates. Derivation of ESG1-deficient ES cells from blastocysts Derivation of ESG1-deficient ES cells from blastocysts Esg1+/-or ESG1-/- mutant female mice were injected with Tamoxifen (10 µg) and Depo-provera (1 mg) subcutane- ously on the third day of pregnancy. Four days later, embryos in diapause were flushed out of the uterus and cultured on STO feeder cells in four-well plates in Dul- becco's Modified Eagle Medium (DMEM) supplemented with 20% Fetal Bovine Serum (Hyclone), 0.1 mM Non- Essential Amino Acids (Invitrogen), 2 mM L-glutamine (Invitrogen), 50 U/ml Penicillin-Streptomysin (Invitro- gen), and 0.11 mM 2-mercaptoethanol (Invitrogen). After six days, the central mass of each explant was harvested, rinsed in PBS, and placed in a drop of trypsin for a few minutes. The cell mass was collected with a finely drawn- out Pasteur pipette preloaded with medium, ensuring minimal carryover of the trypsin. The cells were gently transfered into a fresh well with 20% FBS-containing medium. The resulting primary ES cell colonies were indi- vidually passaged into wells of four-well plates containing Targeted d Figure 4 ESG1 Oct3/4 CDK4 +/- -/- ESG1 101 100 102 103 104 105 101 100 102 103 104 105 ESG1-null ES cell Oct4 Nanog A. B. ESG1 Oct3/4 CDK4 +/- -/- ESG1 101 100 102 103 104 105 101 100 102 103 104 105 ESG1-null ES cell Oct4 Nanog A. A. B. p y g Gene expression analyses of ESG1-null ES cells. A) DNA microarray analyses. Total RNA from wild-type ES cells and ESG1-null ES cells were labeled with Cy3 and Cy5, respectively. Samples were hybridized to Agilent Mouse Developmental Microarrays. The averages of two independent clones are shown. B) Western blot analyses. Cell lysates from ESG1+/- and ESG1-/- cells were examined for expression of ESG1, Oct3/4 and CDK4 with immunoblotting. Cell Signaling) were used to detect antibody binding. We visualized bound antibody with an ECL Western Blotting Detection System (Amersham). construct pDONR-pH34, the resulting PCR product was subcloned into pDONR201 (Invitrogen). pDONR-pH34 was interacted with pDEST17 (Invitrogen) by LR recombi- nation. After introduction of the resulting expression vec- tor pDEST17-pH34 into BL21-AI E. coli (Invitrogen), recombinant protein production was induced according to the manufacture's protocol. Histidine-tagged ESG1 was purified using Ni-nitrilotriacetic acid agarose (Qiagen) under denaturing conditions in the presence of 8 M urea. After dialysis against 6 M urea, the recombinant proteins were injected into New Zealand White rabbits to generate anti-ESG1 polyclonal antibodies. Targeted d Figure 4 Cell numbers were determined with a Coulter counter at 2, 4, and 6 days. Data of +/- and -/- cells are shown as aver- ages and standard deviations of three independent clones. C) A section of teratoma derived from ESG1-null ES cells (hematox- ylin & eosin staining). TCAGTTAT -3') and pH34L-9000 (5'-GATAAGCTGCT- GCCAAAAGACAAG -3') primers. The probe hybridized to an 11.5 kbp band from the wild-type locus, a 12.5 kbp band from the β-geo locus, and a 9.5 kbp band from the HygR locus. Generation of anti-ESG1 polyclonal antibodies The coding sequence of Esg1 was amplified by PCR with the pH34-gw-s (5'- AAAAAGCAGGCTGGATGATGGT- GACCCTCGTGA-3') and pH34-gw-as (5'- AGAAAGCT- GGGTCTGCATCCAGGTCGGAGACA-3') primers. To Page 7 of 9 (page number not for citation purposes) TCAGTTAT -3') and pH34L-9000 (5'-GATAAGCTGCT- GCCAAAAGACAAG -3') primers. The probe hybridized to an 11.5 kbp band from the wild-type locus, a 12.5 kbp band from the β-geo locus, and a 9.5 kbp band from the HygR locus. Generation of anti-ESG1 polyclonal antibodies The coding sequence of Esg1 was amplified by PCR with the pH34-gw-s (5'- AAAAAGCAGGCTGGATGATGGT- GACCCTCGTGA-3') and pH34-gw-as (5'- AGAAAGCT- GGGTCTGCATCCAGGTCGGAGACA-3') primers. To TCAGTTAT -3') and pH34L-9000 (5'-GATAAGCTGCT- GCCAAAAGACAAG -3') primers. The probe hybridized to an 11.5 kbp band from the wild-type locus, a 12.5 kbp band from the β-geo locus, and a 9.5 kbp band from the HygR locus. Generation of anti-ESG1 polyclonal antibodies The coding sequence of Esg1 was amplified by PCR with the pH34-gw-s (5'- AAAAAGCAGGCTGGATGATGGT- GACCCTCGTGA-3') and pH34-gw-as (5'- AGAAAGCT- GGGTCTGCATCCAGGTCGGAGACA-3') primers. To Generation of anti-ESG1 polyclonal antibodies The coding sequence of Esg1 was amplified by PCR with the pH34-gw-s (5'- AAAAAGCAGGCTGGATGATGGT- GACCCTCGTGA-3') and pH34-gw-as (5'- AGAAAGCT- GGGTCTGCATCCAGGTCGGAGACA-3') primers. To Page 7 of 9 (page number not for citation purposes) BMC Developmental Biology 2006, 6:11 http://www.biomedcentral.com/1471-213X/6/11 Gene expression analyses of ESG1-null ES cells Figure 6 Gene expression analyses of ESG1-null ES cells. A) DNA microarray analyses. Total RNA from wild-type ES cells and ESG1-null ES cells were labeled with Cy3 and Cy5, respectively. Samples were hybridized to Agilent Mouse Developmental Microarrays. The averages of two independent clones are shown. B) Western blot analyses. Cell lysates from ESG1+/- and ESG1-/- cells were examined for expression of ESG1, Oct3/4 and CDK4 with immunoblotting. ESG1 101 100 102 103 104 105 101 100 102 103 104 105 Wild-type ES cell ESG1-null ES cell Oct4 Nanog A. B. Authors' contributions ( ) 14. Western P, Maldonado-Saldivia J, van den Bergen J, Hajkova P, Saitou M, Barton S, Surani MA: Analysis of Esg1 Expression in Pluripo- tent Cells and the Germline Reveals Similarities with Oct4 and Sox2 and Differences Between Human Pluripotent Cell Lines. Stem Cells 2005, 23(10):1436-1442. HA carried out the phenotypic studies of ESG1 knockout mice. KI determined the chromosomal localizations of the ESG1 gene and pseudogenes and constructed the tar- geting vector. TI carried out mouse embryo manipulation. MM and MN carried out ES cell culture. SY conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript. ( ) 15. Kanatsu-Shinohara M, Inoue K, Lee J, Yoshimoto M, Ogonuki N, Miki H, Baba S, Kato T, Kazuki Y, Toyokuni S, Toyoshima M, Niwa O, Oshimura M, Heike T, Nakahata T, Ishino F, Ogura A, Shinohara T: Generation of pluripotent stem cells from neonatal mouse testis. Cell 2004, 119(7):1001-1012. 16. Gibson TJ, Thompson JD, Heringa J: The KH domain occurs in a diverse set of RNA-binding proteins that include the antiter- minator NusA and is probably involved in binding to nucleic acid. FEBS Lett 1993, 324(3):361-366. Western blot After preparation of ES cell extracts with M-Per (Pierce), cellular proteins were separated on sodium dodecyl sul- fate (SDS)-14% polyacrylamide gels and transferred to nitrocellulose membranes (Millipore). Membranes were incubated with anti-ESG1 (1/500 dilution), anti-Oct3/4 (1/500; Santa Cruz Biotechnology), anti-CDK4 (1/200; Santa Cruz Biotechnology), and anti-GFP (1/1000; MBL) primary antibodies. Horseradish peroxidase-conjugated anti-rabbit and anti-mouse immunoglobulins (1/5000; Page 8 of 9 (page number not for citation purposes) Page 8 of 9 (page number not for citation purposes) Microarrays 10. Takahashi K, Mitsui K, Yamanaka S: Role of ERas in promoting tumour-like properties in mouse embryonic stem cells. Nature 2003, 423(6939):541-545. Total RNA from wild-type ES cells and ESG1-/- ES cells was labeled with Cy3 and Cy5, respectively. The samples were hybridized to a Mouse Development Microarray (Algi- lent) according to the manufacturer's protocol. Arrays were scanned with a G2565BA Microarray Scanner System (Agilent). Hybridization was repeated with two independ- ent clones. Data were analyzed with GeneSprings software (Silico Genetics). ( ) 11. Mitsui K, Tokuzawa Y, Itoh H, Segawa K, Murakami M, Takahashi K, Maruyama M, Maeda M, Yamanaka S: The Homeoprotein Nanog Is Required for Maintenance of Pluripotency in Mouse Epi- blast and ES Cells. Cell 2003, 113(5):631-642. ( ) 12. Maruyama M, Ichisaka T, Nakagawa M, Yamanaka S: Differential roles for sox15 and sox2 in transcriptional control in mouse embryonic stem cells. J Biol Chem 2005, 280(26):24371-24379. 13. Astigiano S, Barkai U, Abarzua P, Tan SC, Harper MI, Sherman MI: Changes in gene expression following exposure of nulli-SCCl murine embryonal carcinoma cells to inducers of differenti- ation: characterization of a down-regulated mRNA. Differen- tiation 1991, 46(1):61-67. http://www.biomedcentral.com/1471-213X/6/11 BMC Developmental Biology 2006, 6:11 STO feeder cell layers. Thereafter, cells were expanded by trypsinization of the entire culture. 9. Tokuzawa Y, Kaiho E, Maruyama M, Takahashi K, Mitsui K, Maeda M, Niwa H, Yamanaka S: Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic stem cell self-renewal and mouse development. Mol Cell Biol 2003, 23(8):2699-2708. 10. References References 1. Evans MJ, Kaufman MH: Establishment in culture of pluripoten- tial cells from mouse embryos. Nature 1981, 292(5819):154-156. ( ) 22. Meiner VL, Cases S, Myers HM, Sande ER, Bellosta S, Schambelan M, Pitas RE, McGuire J, Herz J, Farese RVJ: Disruption of the acyl- CoA:cholesterol acyltransferase gene in mice: evidence sug- gesting multiple cholesterol esterification enzymes in mam- mals. Proc Natl Acad Sci U S A 1996, 93(24):14041-14046. ( ) 22. Meiner VL, Cases S, Myers HM, Sande ER, Bellosta S, Schambelan M, Pitas RE, McGuire J, Herz J, Farese RVJ: Disruption of the acyl- CoA:cholesterol acyltransferase gene in mice: evidence sug- gesting multiple cholesterol esterification enzymes in mam- mals. Proc Natl Acad Sci U S A 1996, 93(24):14041-14046. ( ) 2. Martin GR: Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by terato- carcinoma stem cells. Proc Natl Acad Sci U S A 1981, 78(12):7634-7638. ( ) 3. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM: Embryonic stem cell lines derived from human blastocysts. Science 1998, 282(5391):1145-1147. ( ) 3. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM: Embryonic stem cell lines derived from human blastocysts. Science 1998, 282(5391):1145-1147. Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Page 9 of 9 (page number not for citation purposes) Publish with BioMed Central and every scientist can read your work free of charge "BioMed Central will be the most significant development for disseminating the results of biomedical research in our lifetime." Sir Paul Nurse, Cancer Research UK Your research papers will be: available free of charge to the entire biomedical community peer reviewed and published immediately upon acceptance cited in PubMed and archived on PubMed Central yours — you keep the copyright Submit your manuscript here: http://www.biomedcentral.com/info/publishing_adv.asp BioMedcentral Publish with BioMed Central and every scientist can read your work free of charge y , ( ) 4. Chambers I, Smith A: Self-renewal of teratocarcinoma and embryonic stem cells. Oncogene 2004, 23(43):7150-7160. y , ( ) 4. Chambers I, Smith A: Self-renewal of teratocarcinoma and embryonic stem cells. Oncogene 2004, 23(43):7150-7160. y g , ( ) 5. Wobus AM, Boheler KR: Embryonic stem cells: prospects for developmental biology and cell therapy. Physiol Rev 2005, 85(2):635-678. y g ( ) 5. Wobus AM, Boheler KR: Embryonic stem cells: prospects for developmental biology and cell therapy. Physiol Rev 2005, 85(2):635-678. ( ) 6. 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Devel- opment 2003, 130(8):1673-1680. Acknowledgements ( ) 17. Bierbaum P, MacLean-Hunter S, Ehlert F, Moroy T, Muller R: Cloning of embryonal stem cell-specific genes: characterization of the transcriptionally controlled gene esg-1. Cell Growth Differ 1994, 5(1):37-46. g We thank Chihiro Takigawa, Junko Iida, Masako Shirasaka, Yumi Ohuchi, and Megumi Narita for their technical and administrative assistance. We are grateful to Drs. Minoru Ko and Azim Surani for sharing helpful data prior to publication, Dr. Hitoshi Niwa for generously providing various vectors, and Dr. Robert Farese Jr. for his kind gift of the RF8 ES cells. This work was supported in part by research grants from the Ministry of Education, Cul- ture, Sports, Science, and Technology of Japan, the Uehara Memorial Foun- dation, the Naito Foundation, the Sumitomo Research Foundation, the Mitsubishi Foundation. S.Y. was supported by a Toray Science and Technol- ogy Grant (to S.Y.). 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https://openalex.org/W2058666334	https://hal.archives-ouvertes.fr/hal-00771939/file/journal.pone.0049406.pdf	English	null	A Generic Model to Simulate Air-Borne Diseases as a Function of Crop Architecture	PloS one	2,012	cc-by	11,037	To cite this version: Pierre Casadebaig, Gauthier Quesnel, Michel Langlais, Robert Faivre. A Generic Model to Sim- ulate Air-Borne Diseases as a Function of Crop Architecture. PLoS ONE, 2012, 7 (11), pp.1-11. 10.1371/journal.pone.0049406. hal-00771939 Pierre Casadebaig1, Gauthier Quesnel1, Michel Langlais2, Robert Faivre1 1 Unite´ de Biome´trie et Intelligence Artificielle UPR 875, Institut National de la Recherche Agronomique, Centre de Recherches de Toulouse, Castanet-Tolosan, France, 2 Institut de Mathematiques de Bordeaux, Universite´ de Bordeaux, Bordeaux, France 1 Unite´ de Biome´trie et Intelligence Artificielle UPR 875, Institut National de la Recherche Agronomique, Centre de Recherches de 2 Institut de Mathematiques de Bordeaux, Universite´ de Bordeaux, Bordeaux, France HAL Id: hal-00771939 https://hal.science/hal-00771939v1 Submitted on 9 Jan 2013 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. Abstract This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was granted by Agence Nationale de la Recherche (project ARCHIDEMIO, grant ANR-08-STRA-04-07). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Funding: This work was granted by Agence Nationale de la Recherche (project ARCHIDEMIO, grant ANR-08-STRA-04-07) data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. E-mail: pierre.casadebaig@toulouse.inra.fr In a context of pesticide use reduction, alternatives to chemical- based crop protection strategies are needed to control diseases. Before the pesticide era, the canopy structure - resulting from the plant architecture and crop management - was the main natural lever to try to control disease development in cultivated fields. An update of this expertise is proposed through a research project focused on four architecturally contrasted systems and their main associated diseases: pisum-aschochyta blight, potato-late blight, vineyard-powdery mildew and yam-anthracnose systems. The first two systems were choosen as case studies for this paper. Knowledge in epidemiology tends to be very specific to the studied system. Modeling can help identify and share similarities between systems to unravel crop and pathogen interactions on a wider range of systems. Later, as a more operational step, modeling could be used to quantify how the canopy structure affects disease outbreaks and to identify architectural traits or cropping practices that lower disease severity. Provided that the model embeds genetic information (genetic basis of phenotypic parameters) it may later become a tool to design adapted genotypes [6,7]. Abstract In a context of pesticide use reduction, alternatives to chemical-based crop protection strategies are needed to control diseases. Crop and plant architectures can be viewed as levers to control disease outbreaks by affecting microclimate within the canopy or pathogen transmission between plants. Modeling and simulation is a key approach to help analyze the behaviour of such systems where direct observations are difficult and tedious. Modeling permits the joining of concepts from ecophysiology and epidemiology to define structures and functions generic enough to describe a wide range of epidemiological dynamics. Additionally, this conception should minimize computing time by both limiting the complexity and setting an efficient software implementation. In this paper, our aim was to present a model that suited these constraints so it could first be used as a research and teaching tool to promote discussions about epidemic management in cropping systems. The system was modelled as a combination of individual hosts (population of plants or organs) and infectious agents (pathogens) whose contacts are restricted through a network of connections. The system dynamics were described at an individual scale. Additional attention was given to the identification of generic properties of host-pathogen systems to widen the model’s applicability domain. Two specific pathosystems with contrasted crop architectures were considered: ascochyta blight on pea (homogeneously layered canopy) and potato late blight (lattice of individualized plants). The model behavior was assessed by simulation and sensitivity analysis and these results were discussed against the model ability to discriminate between the defined types of epidemics. Crop traits related to disease avoidance resulting in a low exposure, a slow dispersal or a de-synchronization of plant and pathogen cycles were shown to strongly impact the disease severity at the crop scale. Citation: Casadebaig P, Quesnel G, Langlais M, Faivre R (2012) A Generic Model to Simulate Air-Borne Diseases as a Function of Crop Architecture. PLoS ONE 7(11): e49406. doi:10.1371/journal.pone.0049406 Editor: Vittoria Colizza, INSERM & Universite Pierre et Marie Curie, France Received February 3, 2012; Accepted October 10, 2012; Published November 30, 2012 Editor: Vittoria Colizza, INSERM & Universite Pierre et Marie Curie, France Editor: Vittoria Colizza, INSERM & Universite Pierre et Marie Curie, France Received February 3, 2012; Accepted October 10, 2012; Published November 30, 2012 Received February 3, 2012; Accepted October 10, 2012; Published November 30, 2012 Copyright: ! 2012 Casadebaig et al. November 2012 \| Volume 7 \| Issue 11 \| e49406 Analysis Essentially, a crop-pathogen system can be viewed as the juxtaposition of individual hosts (plants) and infectious agents (pathogens) whose contacts are restricted through dispersal network; these three elements evolving under a fluctuating climate. By adopting a shared point of view on the crop and the pathogen, this dispersal network is defined by both the crop structure and the pathogen’s dispersal mode. The properties of the generic objects defining the system are here detailed and are the conceptual basis of the model. This conceptual basis merges basic concepts from epidemiology, crop modeling and graph theory. The structure and dynamics of the formal model are presented in the Results section. We assume that an approximation of the geometry rather than a fully detailed 3D plant structure is an acceptable simplification when considering the crop at an integrative level. When dealing with processes at crop scale, such as epidemic development, the underlying complexity could partly be discarded. If complexity is ignored, the model is able to describe the epidemic (using logistic growth for example) but significant interactions between elements of the system will not be highlighted. Merging concepts from basic epidemiology, crop modeling and graph theory Brief history of epidemic models in phytopathology. When observing the evolution of disease severity at the field scale, the crop leaf area is usually logistically destructed by the pathogen. As reviewed by Hau [8], two main approaches assuming different levels of complexity were investi- gated in the bibliography to model this observation: (1) mecha- nistic detailed models dealing with small-scale processes to understand the functioning of the whole system and (2) analytical models, based mostly on differential equations, with a limited number of processes and parameters [9]. The detailed approach tends to be very specific of the system (crop, pathogen species). An important feature of our approach is to build a model suitable for different systems, so we focused on the latter approach, based on simpler dynamics. We have chosen to keep sub-crop scale objects (plants or organs) but discard their geometric attributes (size, shape, spatial position and orientation). The canopy was represented by an object that was simply named functional unit, the scale of this object was relative to the epidemic type. It was also more convenient to describe processes at the level where they are better known for each system. For example, in dense canopy, where epidemics tend to ascend from lower crop layers [20], individual plants are less pertinent than a representation of a mean plant divided in stem segments (phytomers). In patchy epidemics, individual plants get infected and disperse infectious agents in their local neighbourhood. These two epidemic archetypes were taken as a case study. Thus, functional units can represent either a mean plant, individual plants or a sub-part of a plant. The canopy architecture was characterized by a network of functional units. Dynamics of plant and pathogen growth were described on functional units, meaning that a homogeneous behavior of plant tissues was supposed within the unit. In the first models from Vanderplank [9] the evolution of disease severity (y) can be described by the logistic growth rate dy=dt~ry(1{y) as a function of time (t) and an infection rate (r). Such simple growth functions can be used to describe epidemics but not to explain the underlying biological processes [8]. Customized models could be built from this starting point to include biological processes such as the partitioning of the diseased individuals into different compartments (healthy, latent, infectious and removed) [10], namely SEIR models in human epidemiology. Epidemiological Modelling and Crop Architecture also more complicated. In analytical models, parameters such as r in the logistic growth contains information about the contacts underlying the disease transmission process. In this case, random contacts are assumed in the population (random mixing network). Considering non-mobile hosts, the number of contacts that each individual has is considerably smaller than the population size and, in such circumstances, random mixing does not occur [13]. It is thus important to model the contact network that depends not only on the type of pathogen’s dispersion (wind, splashing, …) but also on the host’s spatial arrangement. canopy function and structure to design a model with a generic scope, i.e. based on a single representation for different pathosystems, to explain contrasted dynamics of epidemics. Secondly, as the analysis to understand and use the model would be heavily based on simulation, the model design should minimize computing time by both limiting the complexity and setting an efficient software implementation. Our aim in this paper was to present a model that suited these constraints so it can first be used as a research and teaching tool to promote discussions about the control of diseases. We first present the conceptual design phase (Analysis section) that defines constraints for the following phase of model design and implementation (Results section). Model evaluation is present- ed in the Discussion section before presenting conclusions and perspectives. Representing the canopy structure with a variable scale object. Plant epidemiology [14] and crop modeling [15,16] are very active research topics but approaches that link these two domains are less frequent [2,17,18]. As the case studies for these models range from improving scientific knowledge to engineering, finding an appropriate coupling scale is difficult. For example, the effect of plant architecture on disease dispersal and severity was modeled by coupling a structural plant model (3D ‘‘virtual plant’’) and experimental disease inoculations [19]. This organ-scale model would not be the most appropriate basis for adding dynamics of host or pathogen growth that would fit different crops, because of the overwhelming parameters fit needed. In our case study, we will use modelling concepts from crop models at a broad scale to allow an easier setup of generic features. Introduction In agriculture, growers have to compete with harmful organisms (animal pests, plant pathogens and weeds), collectively called pests [1]. These pests have different ways to injure plants [2] and cause crop losses (quantitative or qualitative damages), causing economic losses. Among pathogens, diseases related to fungi can severely impair plant physiology by lowering light interception or reducing photosynthetic rate. Thus, yield increases in the last decades resulted essentially from genetic progress and massive use of pesticides. However, current practices in crop protection generate a set of issues: (1) economical, as the cost of chemical treatments lowers margins when crop prices or yields are low; (2) durability, as the use of pesticides causes strong environmental risks: some pathogen populations can adapt and gain resistance rapidly (around 4 years); (3) technical, as the integration of alternative methods of crop protection (genetic, chemical, physical, biological and cultural control) is hard to achieve in a farm and finally (4) scientific, as the joint management of plant and pathogen populations in time (crop rotations) and space (landscape structure, [3]) is still a research question (integrated avirulence management, [4,5]). This scientific and operational context set several constraints on the model development. The main objective of this work was to cross the point of view of ecophysiologists and pathologists on the November 2012 \| Volume 7 \| Issue 11 \| e49406 November 2012 \| Volume 7 \| Issue 11 \| e49406 1 PLOS ONE \| www.plosone.org Epidemiological Modelling and Crop Architecture Epidemiological Modelling and Crop Architecture Setting up an experiment to determine the real network of connection is tedious for spore-transmitted infections as relation- ships between individuals are not observable in field. Therefore, the representation of the real network results from an a priori point of view on both crop structure and dispersal mode rather than direct observation. pathosystems were taken as example to test the model genericity: (1) the pisum-aschochyta system [20], where the description of epidemics combines a local spore dispersal (splashing, stem-flow) and organ scale knowledge and (2) the potato late blight system that with long distance wind-dispersal between individualized plants [24]. Two corresponding methods, respectively named ‘‘stem’’ and ‘‘grid’’ are presented here to generate these contrasted canopy archetypes. Model overview Stem method is used to simulate pathosystems presenting homogeneous canopies where epidemics follow a vertical dynamic. In this method, a single plant is represented as a succession of growing stem segments. The hypothesis of an homogeneous crop in the plot (mean plant) allowed aggregation at crop scale. The horizontal plot variability coming from soil, climate or primary inoculum repartition was not considered. In the model, functional units are created sequentially and recursively linked together to mimic stem growth. Thus, each functional unit represents a layer of organs having a similar physiological age over the whole canopy. The dynamic construction of the units was implemented with a finite state automaton (modelled with an Hortel statechart [25], a variant of the UML statechart [26]). In the statechart presented in figure 2, states correspond to units and the transitions between states are conditioned by thermal time accumulation. Different phyllochron values were defined to simulate preformed stem segments (tu is a vector). The unit construction process stops at flowering time (tf ), determined at crop level. This method also defines the relationship between units: each unit is connected to its upper neighbour (linear graph). The epidemiological model consists of individual-based sub- models for pathogen and leaf area dynamics linked together by a network that represents the canopy architecture. Figure 1 shows these two scales, a global one (crop) and a local one (individual hosts referred to as functional units) which interplay by using common attributes. This conception allowed individual host dynamics to be affected by global crop properties that are in turn dependent of individual attributes. Figure 1 presents a static view of the model before the beginning of the simulation. As the simulation progresses, functional units are created and linked together according to the specifications of the connection network. The canopy structure and development processes are detailed in the Unit Construction sub-model. The Functional Units model describes the dynamics of plant (leaf area) and pathogen (leaf area injured) growth at local scale as a set of ordinary differential equations. The environmental effect on pathogen growth is computed in the Environment sub-model. The formal model is deterministic with a discrete time-step (daily). Model’s variables are summarized in table 1. System input variables are climatic measurements (mean air temperature). Input parameters are defined in table 2. Merging concepts from basic epidemiology, crop modeling and graph theory The interaction between host and pathogen growth could also be added to simpler models, as initiated by Jeger [11]. The main hypothesis underneath this model was the potential equilibrium state between host and pathogen populations. Such model design could extend to the integration of different organization scales still without using explicitly spatialized variables [12]. Modeling spore dispersal by using a graph-theory perspective. Early epidemiological models (logistic growth or SEIR-derived) have a long and successful history but are based on population-wide random mixing [13]. In agronomic systems, where canopy results mainly from interaction between cropping practices and the environment, there is often a strong spatial structure of the host population (rows, trellis). In practice, each individual has a finite set of contacts that they can infect. An analogy with graph theory, with hosts as graph nodes and neighbourhood of infection as edges, seems an adapted approach to deal with these limited contacts [13,21]. In our model, a graph, namely the network of connections between functional units, was used to represent the spore dispersal process. If further complexity is added in theoretical models, represent- ing a generic behavior of a pathogen class (fungi) is trickier and less pertinent. Consequently, the computational study of the model is November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 2 Epidemiological Modelling and Crop Architecture Individual dynamics for plant and pathogen growth (Functional Units sub-model) This model relies on two hypotheses: (1) the disease develop- ment does not affect the plant growth rate (ke) nor the potential size (Emax), (2) there is no specific feedback of the disease on natural leaf senescence (DS is disease-independent). ( ) The dynamic of functional units, whether they represent plants or organs, was defined by two interacting equation systems. The first one is related to healthy plant functioning and the second depicts disease development on leaf tissues. These two systems include terms that depend on state variables from linked functional units, allowing to model disease development at the canopy scale. Functional units were mainly described by leaf area because it is a driving variable for light interception (linked to crop productiv- ity) and a physical support for pathogen development. The units also included height and porosity attributes. As in SEIR family models [4,27], total leaf area for each functional unit is divided into distinct compartments. From the plant point of view, we defined expanded (E), senescent (S) and photosynthetically active (A) leaf area. For the pathogen, the healthy (H), infected (L, Latent), infectious (I) and removed (R) compartments. The removed class combined leaf area destruction caused by natural senescence and by disease development. If there is no infection, all the leaf The dynamic of functional units, whether they represent plants or organs, was defined by two interacting equation systems. The first one is related to healthy plant functioning and the second depicts disease development on leaf tissues. These two systems include terms that depend on state variables from linked functional units, allowing to model disease development at the canopy scale. Dynamic of leaf area in disease condition. The diffusion of pathogenic agents (i.e. spores) was not modelled explicitly and is approximated by rates of conversion between the different leaf area compartments. The aim of the pathogen model (eq. 4) was to merge a compartmental model of leaf injury and the host plant growth. This model was adapted from previously developed model on the powdery mildew - vineyard system [29]. In our case, two processes were refined: (1) healthy area was dependent of host growth and was simulated with a basic crop model and (2) both pathogen-induced and natural senescence (DS) contributed to the removal of leaf area from the system. Canopy development (Unit Construction sub-model) Z(t)~ Zmax 1ze{kz:(t{tz) DZ(td)~Z(tdz1){Z(td) 8 < : ð1Þ E(t)~ Emax 1ze{ke:(t{te) DE(td)~E(tdz1){E(td) 8 > < > : ð2Þ S(t)~ Emax 1ze{ks:(t{ts) DS(td)~S(tdz1){S(td) 8 > < > : ð3Þ Z(t)~ Zmax 1ze{kz:(t{tz) DZ(td)~Z(tdz1){Z(td) 8 < : ð1Þ ð1Þ E(t)~ Emax 1ze{ke:(t{te) DE(td)~E(tdz1){E(td) 8 > < > : ð2Þ ð2Þ S(t)~ Emax 1ze{ks:(t{ts) DS(td)~S(tdz1){S(td) 8 > < > : ð3Þ ð3Þ Horizontal lines separate crop attributes from unit specific attributes. Rate- based variables directly linked to state variables were not specified in the table. doi:10.1371/journal.pone.0049406.t001 Emax;Zmax the value of asymptote, representing the potential of the elongation (Zmax), expansion or senescence (Emax) process whether a whole plant (total leaf area, plant height) or an organ (phytomer area, internode length) is considered. These parameters are measurable on plants growing in healthy conditions and without environmental stress. sented by defining asymmetric graphs (D, E, F). For example, graph E is used to simulate long-range dispersal events occurring at a low frequency. Graphs C and F are defined to represent row-structured crops, with a weak inter-row connection for graph F. At the crop scale, this theoretical network of unit connections is parameterized by the adjacency matrix of the corre- sponding graph. The outward edges of each node are weighted to adjust dispersal rate in relation to the leaf area development of the emitting unit (porosity attribute, cf. eq. 6). te; ts; tz the value of the abscissa at the inflection point, representing thermal-time at half the value of Emax or Zmax. Their value was estimated through optimization. We stated that te~tz, meaning that elongation is synchronized with expansion. ke; ks; kz the value of the slope at the inflection point. This value is supposed to be constant among functional units (different between processes) and was optimized. Canopy development (Unit Construction sub-model) In plant models, time scale is frequently coupled with temperature as most physiological processes are temperature- dependent. The concept of thermal time [22] implies that time, as sensed by plants, elapses more rapidly at higher than at lower temperatures in a given range of temperatures [23] and is stopped at a base temperature. It is convenient as dates or rates expressed in thermal time are constant between environments. The main development stages for the crop (emergence, flowering, maturity) are expressed as a sum of degree-days (mean air temperature). As a consequence, calendar time (d, day) and thermal time (t, uC:d) were defined as different mathematical notations. Grid method is used to simulate patchy epidemics, with each unit representing an individual plant. In this case, all units are created simultaneously at the start of the simulation and are linked according to a network of connection. This global network is build from an a priori on the neighbourhood of individual units about both the pathogen dispersal process and the crop structure. This neighbourhood, relatively to individual units, is defined as a deterministic parameter (graph). Examples of possible individual neighbourhoods are presented in figure 3. The strength of the dispersal process was related to the number of plants considered in theses graphs (i.e graphs A vs B and D vs E). Constraints on range or direction of the dispersal process are repre- In the Unit Construction model, the development process of the canopy was represented by the creation of functional units. This structural aspect is distinct from the functioning of each unit. Two Figure 1. General model structure. This summary view represents a hierarchical organization of sub-models which describes objects and processes of the system. Model inputs are shown in dotted frames. Scales (crop and unit) are shown in bold lines. Sub-model Unit Construction acts as a controller for the dynamically created and individual based models Functional Units. doi:10.1371/journal.pone.0049406.g001 Figure 1. General model structure. This summary view represents a hierarchical organization of sub-models which describes objects and processes of the system. Model inputs are shown in dotted frames. Scales (crop and unit) are shown in bold lines. Sub-model Unit Construction acts as a controller for the dynamically created and individual based models Functional Units. doi:10.1371/journal.pone.0049406.g001 November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 3 Epidemiological Modelling and Crop Architecture Table 1. Summary table for variables. Canopy development (Unit Construction sub-model) symbol name unit eq. Tm mean air temperature uC t thermal time uC:d unit number (parameter in ‘‘grid’’ method) - LAI leaf area index m2=m2 t thermal age uC:d 1, 2, 3, 5 Z height m 1 w porosity - 6 r receptivity - 5 h thermal stress - 8 did allo-deposition m2 7 tu:I auto-deposition m2 4 H healthy area m2 4 L latent infected area m2 4 I infectious area m2 4 R removed area m2 4 E expanded area m2 2 A photosynthetically active area m2 S senescent area m2 3 Horizontal lines separate crop attributes from unit specific attributes. Rate- based variables directly linked to state variables were not specified in the table. doi:10.1371/journal.pone.0049406.t001 Table 1. Summary table for variables. symbol name unit eq. Tm mean air temperature uC t thermal time uC:d unit number (parameter in ‘‘grid’’ method) - LAI leaf area index m2=m2 t thermal age uC:d 1, 2, 3, 5 Z height m 1 w porosity - 6 r receptivity - 5 h thermal stress - 8 did allo-deposition m2 7 tu:I auto-deposition m2 4 H healthy area m2 4 L latent infected area m2 4 I infectious area m2 4 R removed area m2 4 E expanded area m2 2 A photosynthetically active area m2 S senescent area m2 3 Horizontal lines separate crop attributes from unit specific attributes. Rate- based variables directly linked to state variables were not specified in the table. doi:10.1371/journal.pone.0049406.t001 area is thus removed through natural senescence. At each time- step, the identity HzLzI~A~E{R holds to maintain leaf area conservation (as for biomass or energy conservation). Table 1. Summary table for variables. Table 1. Summary table for variables. Dynamics of plant elongation, leaf expansion and senescence. Stem height elongation (eq. 1), leaf area expansion (eq. 2) and leaf senescence (eq. 3) rates were modeled with a symmetric logistic equation driven by thermal time t and 3 parameters. This formalism is frequently used at the crop scale in ecophysiology. It was previously adapted at organ-scale [28]. This approach is similar to ours when considering an analogy between organs and functional units. Individual dynamics for plant and pathogen growth (Functional Units sub-model) It was also assumed that the natural senescence process removed leaf area independently from its health status (infected or infectious). It was a way to model disease resistance via avoidance, as leaf area can be removed Functional units were mainly described by leaf area because it is a driving variable for light interception (linked to crop productiv- ity) and a physical support for pathogen development. The units also included height and porosity attributes. As in SEIR family models [4,27], total leaf area for each functional unit is divided into distinct compartments. From the plant point of view, we defined expanded (E), senescent (S) and photosynthetically active (A) leaf area. For the pathogen, the healthy (H), infected (L, Latent), infectious (I) and removed (R) compartments. The removed class combined leaf area destruction caused by natural senescence and by disease development. If there is no infection, all the leaf November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 4 Epidemiological Modelling and Crop Architecture Table 2. Summary table for parameters. symbol name stem grid uncertainty unit eq. Individual dynamics for plant and pathogen growth (Functional Units sub-model) Tb base temperature 0 4 uC tu phyllochrons 70 ; 50 NA uC .d tf crop thermal time to flowering 1500 2000 uC .d Zmax potential height of unit 1 1 m 1 kz increase of elongation rate 0.01 0.015 uC21.d21 1 tz unit / crop thermal time for Zmax=2 300 1000 uC.d 1 Emax potential area of unit 1 1 m2 2 ke increase of expansion rate 0.01 0.015 uC21.d21 2 te unit / crop thermal time for Emax=2 300 1000 uC.d 2 ks increase of senescence rate 0.03 0.06 uC21.d21 3 ts unit / crop thermal time for Emax=2 1000 2500 [70; 130] % uC.d 3 ip duration of infectious period 10 10 [1;15] d 4 lp duration of latency period 3 5 [1;15] d 4 I0 quantity of inoculation 0.01 0.01 [0.01; 0.3] m2 Di day of disease initiation 50 50 [30; 140] d tn scalar for between-unit infection rate 0.4 0.5 [0.05; 1] - 4 tu scalar for within-unit infection rate 0.6 0.5 [0.05; 1] - 4 tr shape of receptivity function 500 500 [100; 800] uC.d 5 kr slope of receptivity function 0.05 0.01 uC21.d21 5 w0 scalar for porosity level 0.8 0.9 [0.7; 1] m21 6 Tio optimal temperature for infection 25 25 [10;35] uC 8 Tiw shape parameter of infection response to temperature 2.2e23 2.2e23 - 8 Horizontal lines separate the parameters related to canopy, pathogen and environment. Columns ‘‘stem’’ and ‘‘grid’’ indicate the value of parameters for the specific use-cases. Column ‘‘uncertainty’’ indicates the range of selected parameters in sensitivity analysis (in % when the meaning of the parameter is different in both models). doi:10.1371/journal.pone.0049406.t002 Horizontal lines separate the parameters related to canopy, pathogen and environment. Columns ‘‘stem’’ and ‘‘grid’’ indicate the value of parameters for the specific use-cases. Column ‘‘uncertainty’’ indicates the range of selected parameters in sensitivity analysis (in % when the meaning of the parameter is different in both models). doi:10.1371/journal.pone.0049406.t002 before it became infectious. For com i l before it became infectious. For computing simplicity, these periods are not simulated as a time-lag model, meaning that a fraction of infected area becomes infectious from day one after infection (with a rate of 1=lp). DH~DE{ tu:I: H E :rd:hd ! " { Did: H E :rd:hd ! " { DS: H A ! " DL~ tu:I: H E :rd:hd ! " z Did: H E :rd:hd ! Individual dynamics for plant and pathogen growth (Functional Units sub-model) " { 1 lp :L ! " { DS: L A ! " DI~ 1 lp :L ! " { 1 ip :I ! " { DS: I A ! " DR~ 1 ip :I ! " zDS 8 > > > > > > > > > > > > < > > > > > > > > > > > > : ð4Þ Disease regulation by crop properties. Receptivity, namely the level of ontogenic resistance of the plant tissues, affects local and distant infection rates. Equation 5 presents an example of decreasing ontogenic resistance with the unit aging and showing an increase in the rate of infection on the onset of leaf senescence (rdw1). Ontogenic resistance was modeled as a pathosystem-specific function [30] as tissues could get less resistant to infection with aging (pisum-aschochyta [31], potato late blight [32]) or, conversely, could become nearly immune in older organs (vitis- powdery mildew [33]). Biotrophic or necrotrophic behaviours were partly reproduced by this function. ð4Þ Disease initiation. In the model, the primary infection event occurs at a pre-defined time (at day Di) and a fraction of healthy area (I0) is converted into the infected state. Depending on the epidemic archetype, either all units (for the ‘‘Stem’’ model) or a fixed proportion of the existing units at the time of the infection (‘‘Grid’’ model) are affected. Disease’s development. The dynamic for healthy area (DH) is given by the host growth (DE) minus the area infected from a local source minus the area infected from the neighbour- hood minus the area that became senescent. In consequence, two parameters acting as rates of infection drive the disease progression in the unit (auto-deposition, tu) and between neighbouring units (allo-deposition, tn). The other terms in equation 4 are either regulation function (rd,hd,wd) or normalizing terms (area in the compartment divided by total area). The disease development is also a function of a latency period (lp) and an infectious period (ip). The attribute wd of unit porosity integrates the architectural characteristics (e.g leaf area orientation, leaf surface properties) that are assumed to impact disease transmission and which are not explicitly modeled. Porosity is approximated (Eq. 6) by leaf area density (area/height) weighted by parameter w0. This regulation through porosity impacts the rate of infection from neighbouring units. Individual dynamics for plant and pathogen growth (Functional Units sub-model) r(td)~1{ 1 1zexp{kr:(t{tr) ð5Þ ð5Þ November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org PLOS ONE \| www.plosone.org 5 Epidemiological Modelling and Crop Architecture Figure 2. Hortel statechart for the ‘‘stem’’ method. Hortel statechart for semi-determinate development of plant structure. The conditions for building a new unit (i.e thermal time above a threshold) are specified in transition between states and represents phyllochrons (tu(n)) or crop flowering time (tf ). doi:10.1371/journal.pone.0049406.g002 Figure 2. Hortel statechart for the ‘‘stem’’ method. Hortel statechart for semi-determinate development of plant structure. The conditions for building a new unit (i.e thermal time above a threshold) are specified in transition between states and represents phyllochrons (tu(n)) or crop flowering time (tf ). doi:10.1371/journal.pone.0049406.g002 Figure 2. Hortel statechart for the ‘‘stem’’ method. Hortel statechart for semi-determinate development of plant structure. The conditions for building a new unit (i.e thermal time above a threshold) are specified in transition between states and represents phyllochrons (tu(n)) or crop flowering time (tf ). doi:10 1371/journal pone 0049406 g002 [34]. To face this challenge, we formulated two hypotheses: (1) the individual-based compartment models (Eq. 4 and 7) are functions of crop-scale attributes, to make individual dynamics sensitive to global conditions and (2) the number of connections between units depends on the pathogen dispersal mode. The outgoing infection from a unit Dod is modeled as multiplicative effect of infected area and unit porosity (wd). For each unit, the incoming infection (Did) is modeled as the sum of neighbouring emissions (cf. fig. 3.). Input wd~w0: Ed{Rd Zd ð6Þ ð6Þ Software implementation and data analysis This result is interpreted by the combination of two effects: a high host growth rate (caused by a favorable oceanic climate) and a weak disease transmission rate between plant nodes (tn~0:3). Refining the model prediction at this scale will require additional experiments on controlled conditions (e.g. on leaf receptivity, [31]). Data analysis. The sensitivity analysis and data visualization was performed with R core software [38] with R packages sensitivity and ggplot2 [39]. Global sensitivity analysis aims at explaining the variability of model outputs with respect to the input parameter uncertainties. It is based on intensive use of computer simulations. After setting an experimental design of parameter values, model outputs are evaluated for each parameter set and sensitivity indices are computed. Various methods of global sensitivity analysis exist [40]. To avoid the m10 model evaluations of a full factorial design (10 parameters with each m modalities), we used the extended- FAST method [41] (size 10m, with m~1000) to perform the sensitivity analysis. It allows to evaluate a first-order sensitivity index (influence of only one parameter on the output) and a total Data analysis. The sensitivity analysis and data visualization was performed with R core software [38] with R packages sensitivity and ggplot2 [39]. Global sensitivity analysis aims at explaining the variability of model outputs with respect to the input parameter uncertainties. It is based on intensive use of computer simulations. After setting an experimental design of parameter values, model outputs are evaluated for each parameter set and sensitivity indices are computed. Various methods of global sensitivity analysis exist [40]. To avoid the m10 model evaluations of a full factorial design (10 parameters with each m modalities), we used the extended- FAST method [41] (size 10m, with m~1000) to perform the sensitivity analysis. It allows to evaluate a first-order sensitivity index (influence of only one parameter on the output) and a total Potato late blight. In this case, the model parameters intend to represent a spatial epidemic, where random primo-infection generates primary foci which in turn infect plants in their neighbourhood [24]. It is not clear how to infer the connection network from the real system. On one hand, observation of patches of infection means a short-range diffusion process (new infections are more likely to occur near a previously infected plant). On the other hand, a long-range dispersal is often observed [47]. Qualitative model evaluation A strong hypothesis is that the model genericity is fully supported by the parameter set (cf. table 2) and the individual graph generating the global connection network (cf. figures 2 and 3). This hypothesis was tested here by running the model with two sets of parameters. The first set of parameter values characterizes a vertical epidemic in a homogeneously layered canopy whereas the second represents a spatial epidemic in a field (lattice of individual plants). These parameter sets will be later referred to as vertical or spatial epidemics. In our approach, the model has been built from a conceptual point of view to get gradually closer to the biological reality. In this sense, the parameter estimation on real data was not our priority in the development of the model. Nevertheless, some parameters are quite easy to estimate, especially those related to phenology as they are well studied for a number of crops (e.g. in pea [42,43], potato [44,45]). In the present work, parameters were not estimated with a numerical optimization method, but were defined so that the epidemic dynamics are meaningful according to the pathologists’ knowledge. architecture impact on transmission. Predicting the population dynamics from the behaviour of individuals is a challenge when modeling epidemics transmission. Figure 3. Individual neighbourhood for the grid method. Nodes are individual plants and edges are pathogen dispersal processes. Local dispersion graphs are defined from a individual unit point of view (bold circle) and are used to infer the global network of connections between all functional units at the crop scale. doi:10.1371/journal.pone.0049406.g003 Figure 3. Individual neighbourhood for the grid method. Nodes are individual plants and edges are pathogen dispersal processes. Local dispersion graphs are defined from a individual unit point of view (bold circle) and are used to infer the global network of connections between all functional units at the crop scale. doi:10.1371/journal.pone.0049406.g003 November 2012 \| Volume 7 \| Issue 11 \| e49406 November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 6 Epidemiological Modelling and Crop Architecture Epidemiological Modelling and Crop Architecture and output disease flows are not synchronized: emissions are computed at the next time-step (the day after) in the target units. sensitivity index (influence of one parameter alone and in interaction with the others parameters). These two indices correspond, in the classical variance analysis, to the principal effect and the total (principal+interactions) effect. dod~tn:I:wd did~ P n dod{1 ( ð7Þ ð7Þ Software implementation and data analysis a varying disease score upon the canopy height, caused by the development of ascochyta blight on pea fields [20]. This observed behaviour is assumed to be a consequence of two processes: a frequent disease initiation on lower leaves (older and more receptive) and a local dispersion to upper nodes [31]. The bell shape of the disease profiles is interpreted as a gradient of age-based (ontogenic) resistance, as upper and younger leaves are more resistant to infection and lower leaves could become senescent before being fully infected. The model outputs are presented in figure 4. They display a classical behaviour as the disease starts developing firstly on lower nodes, moving upwards with time (figure 4, lower graph) [46]. Even if individual unit dynamics are rarely observed in field, the upper section of figure 4 indicates that after the beginning of infection (t~50), the system was perturbed to time about t~100 before returning to a healthy state. This result is interpreted by the combination of two effects: a high host growth rate (caused by a favorable oceanic climate) and a weak disease transmission rate between plant nodes (tn~0:3). Refining the model prediction at this scale will require additional experiments on controlled conditions (e.g. on leaf receptivity, [31]). Ascochyta blight on pea. The stake was to reproduce ‘‘infection profiles’’, i.e. a varying disease score upon the canopy height, caused by the development of ascochyta blight on pea fields [20]. This observed behaviour is assumed to be a consequence of two processes: a frequent disease initiation on lower leaves (older and more receptive) and a local dispersion to upper nodes [31]. The bell shape of the disease profiles is interpreted as a gradient of age-based (ontogenic) resistance, as upper and younger leaves are more resistant to infection and lower leaves could become senescent before being fully infected. The model outputs are presented in figure 4. They display a classical behaviour as the disease starts developing firstly on lower nodes, moving upwards with time (figure 4, lower graph) [46]. Even if individual unit dynamics are rarely observed in field, the upper section of figure 4 indicates that after the beginning of infection (t~50), the system was perturbed to time about t~100 before returning to a healthy state. Impact of environmental stress on plant or pathogen dynamics (Environment sub-model) The Crop Climate model aimed to compute environmental impacts on pathogen development. At this step, only the temperature effect on disease development is modeled (Eq. 8). This factor was introduced as a multiplicative term that modulates disease transmission rates within and between units (Eq. 4). This bell-shape function depends on the crop mean temperature (Tm) and two parameters: the optimum temperature for pathogen growth (Tio) and the width of the response at this optimum temperature (Tiw). The parameter T0~1 was defined as a normalizing term to allow dimensional consistency. hd~1{Tiw: (Tm{Tio)2 T2 0 ð8Þ ð8Þ The first step concerning the model evaluation was to reproduce real behaviours, those that pathologists observe in field conditions. The output variable used to evaluate the model and perform a sensitivity analysis is the proportion of leaf area removed by the sole effect of disease (1=ip:I), integrated over time (sowing to harvest) and space (units). Software implementation and data analysis Software implementation. The model development and simulations rely on the VLE software platform (Virtual Laboratory Environment) [35] and the INRA RECORD platform [36]. This platform implements a mathematical specification (DEVS, Dis- crete EVent system Specification [37]) that was originally developed for research in modeling and simulation independently of the domain of application. The main feature of the VLE platform is that it proposes software sub-formalisms between the abstract DEVS specifications and common mathematical formal- isms in environmental sciences (e.g ODE, Hortel statechart, cellular automatons). Consequently, most of the formal model was implemented by using these extensions rather than manipulating underlying DEVS formalism and making it possible to integrate heterogeneous formalisms in the same model. Dynamic creation of model structure. Following the class/object paradigm, the Functional units model is defined as a class model whose instantiation depends on the model graph structure. Depending on the pathosystem, instantiation is static (at plant emergence) or dynamic during simulation (following the plant development); these new units interact according to the specification of the connection network. This allowed dissociating the biological representation from its consequence on the graph and simulation. Software implementation. The model development and simulations rely on the VLE software platform (Virtual Laboratory Environment) [35] and the INRA RECORD platform [36]. This platform implements a mathematical specification (DEVS, Dis- crete EVent system Specification [37]) that was originally developed for research in modeling and simulation independently of the domain of application. The main feature of the VLE platform is that it proposes software sub-formalisms between the abstract DEVS specifications and common mathematical formal- isms in environmental sciences (e.g ODE, Hortel statechart, cellular automatons). Consequently, most of the formal model was implemented by using these extensions rather than manipulating underlying DEVS formalism and making it possible to integrate heterogeneous formalisms in the same model. Dynamic creation of model structure. Following the class/object paradigm, the Functional units model is defined as a class model whose instantiation depends on the model graph structure. Depending on the pathosystem, instantiation is static (at plant emergence) or dynamic during simulation (following the plant development); these new units interact according to the specification of the connection network. This allowed dissociating the biological representation from its consequence on the graph and simulation. Ascochyta blight on pea. The stake was to reproduce ‘‘infection profiles’’, i.e. Sensitivity analysis We have seen in the former section that it was possible to reproduce observed real behaviors using specific sets of parameter values. Sensitivity analysis can be useful to determine which parameters are the most influential on the variability of the model output, which are those having no influence (precise knowledge on these parameters may not be not necessary), which are those interacting together. It can help the modeler to check if the model respects the a priori knowledge on relative effect of parameters and their interactions. This analysis can also help set research priorities. For example, to help decide which experiments are needed to improve the precision on the estimation of some parameters or to identify regions of the parameter space with a With our model, a quantitative proportion of injured leaf area is used as output data, as pest injuries are usually observed on quantitative scales. Observations to discriminate between patho- gen-induced or senescence-induced removal of leaf area is a very difficult task, if observation is not performed continuously. This modeling approach permits to estimate the respective proportion of leaf area removal caused by these two processes. Protocols for characterization of pest injuries in the field [48] should be Figure 5. Spatial epidemics: disease development in a lattice of individualised plants. The four maps show the disease severity (grey scale) on grids of 50650 plants at the end of simulation. Disease initiation was done at random on 1 percent of total cell number for both cases. These maps differed in the parameterization of the connection network between plants (cf. figure 3): upper maps resulted from symmetrical graphs (i.e. graphs A and B in figure 3), lower maps resulted from asymmetrical graphs (D and E); right and left maps resulted from a variation in the number of plants considered for local dispersion (4 and 8). doi:10.1371/journal.pone.0049406.g005 Figure 5. Spatial epidemics: disease development in a lattice of individualised plants. The four maps show the disease severity (grey scale) on grids of 50650 plants at the end of simulation. Disease initiation was done at random on 1 percent of total cell number for both cases. These maps differed in the parameterization of the connection network between plants (cf. figure 3): upper maps resulted from symmetrical graphs (i.e. Epidemiological Modelling and Crop Architecture Epidemiological Modelling and Crop Architecture Figure 4. Vertical epidemics: disease development in a homogeneously layered canopy. The four upper graphs show individual dynamics of functional units, with each graph corresponding to a state of infection. The output variables are relative to the total leaf area in the unit. The lower graph presents the evolution of ‘‘infection profiles’’, namely the severity of the disease in relation to the height (stem nodes) in the crop. doi:10.1371/journal.pone.0049406.g004 Figure 4. Vertical epidemics: disease development in a homogeneously layered canopy. The four upper graphs show individual dynamics of functional units, with each graph corresponding to a state of infection. The output variables are relative to the total leaf area in the unit. The lower graph presents the evolution of ‘‘infection profiles’’, namely the severity of the disease in relation to the height (stem nodes) in the crop. doi:10.1371/journal.pone.0049406.g004 addressed in order to evaluate the predictive quality of our modeling. addressed in order to evaluate the predictive quality of our modeling. the dispersal neighbourhood. Four examples were chosen from graphs in figure 3. The asymmetry of some graphs, used as connection network, provide interesting heterogeneity at the canopy scale. This parameterization could represent epidemics in heterogeneous fields (e.g. prevailing winds, asynchronous plant emergence). These results illustrate the plasticity we achieved in simulation rather than a formal sensitivity analysis on this connection network. Software implementation and data analysis Thus, the simulations showed in figure 5 aim at exploring the sensitivity of the epidemic process to changes in size or range in November 2012 \| Volume 7 \| Issue 11 \| e49406 7 PLOS ONE \| www.plosone.org Epidemiological Modelling and Crop Architecture November 2012 \| Volume 7 \| Issue 11 \| e49406 November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 8 Epidemiological Modelling and Crop Architecture Epidemiological Modelling and Crop Architecture distribution within this range (figure 6, column ‘‘uncertainty’’). The Extended-FAST method [41] was used to explore parameter space and compute sensitivity indices. experiments would be necessary to develop an intuitive under- standing of network-based epidemics and the effect of network structure [13]. Thereby, to simulate systems with a high number of units and a nearly-complete connection graph (e.g. vineyards) will be highly demanding in computing time: statistics on connection network on real pathosystems would help reduce this uncertainty. A first step, to improve the present model, is to focus on the climatic environment at the local scale (microclimate or phyllo- climate [50]) as this local heterogeneity could probably explain finer host\|pathogen\|environment interactions. The modular model conception makes this improvement easy to take into account. Additionally, environmental stresses are most probably not sensed independently by plant (or organs). An interesting model combining the action of temperature and wetness on pathogen response, proposed by Duthie [51], could be considered to integrate the joint effect of stresses for a range of foliar pathogens. Cropping practices (e.g. pruning, lodging) that dramatically affect the crop structure are not yet included in the present model and should improve its relevance for practical applications. Whatever the model, the three most influential parameters on disease severity are the disease initiation day, the latency period and the onset of senescence. All three parameters are time-related. This analysis points out that avoidance strategy strongly explains disease severity: whether this avoidance came from a low exposure (late inoculum arrival), a slow disease dispersal (long latency period) or a de-synchronisation of plant and pathogen cycle (early plant senescence). The high impact of the disease initiation day parameter on the severity suggests that the control of primary inoculum production is a key point for crop protection. This point should be addressed in connection with management strategies, i.e. interaction between tillage and crop sequence to manage infected stubble, [49]. As expected, estimated sensitivity indices highlight the strong impacts of both the network transmission and senescence parameters. The impact of senescence on severity is higher in spatial than in vertical epidemics. This is interpreted as the consequence of the synchronisation of host development in the spatial model. Interestingly, all parameters interacted more tightly with each other in spatial epidemics (larger total index). About practical development. Acknowledgments The authors warmly thank agronomists and phytopathologists implicated in the project for their fruitful collaboration. The authors also specially thanks Jean-Noe¨l Aubertot for his helpful review of the manuscript. Conclusions As stated before, the developed model lies in the research domain. It needs more modeling iterations with phytopathologists to be able to produce outputs that can be used for agricultural applications. Nevertheless, the modeling process raises interesting questions that could be scrutinised with experimental approaches: the disease impact on plant senescence, previously viewed as an additional process to the model, was finally represented as a simple competition for leaf area. Simulation plays a great role to help grasping the functioning of agricultural systems where direct observations are difficult and tedious, providing a sound balance between real and virtual experiments. Author Contributions About conceptual development. A consequent part of the model adaptation to specific systems was achieved through the structure of unit network. As its impact cannot be directly evaluated through classical sensitivity analysis, additional virtual About conceptual development. A consequent part of the model adaptation to specific systems was achieved through the structure of unit network. As its impact cannot be directly evaluated through classical sensitivity analysis, additional virtual Conceived and designed the experiments: PC RF ML. Performed the experiments: PC GQ. Analyzed the data: PC RF. Contributed reagents/ materials/analysis tools: PC GQ RF. Wrote the paper: PC RF GQ. Designed the modeling and simulation platform: GQ. Conceived and designed the experiments: PC RF ML. Performed the experiments: PC GQ. Analyzed the data: PC RF. Contributed reagents/ materials/analysis tools: PC GQ RF. Wrote the paper: PC RF GQ. Designed the modeling and simulation platform: GQ. Epidemiological Modelling and Crop Architecture The modeling of two types of epidemics within one single model structure was a first step demonstrating that the defined objects (functional units and a network of connection) had sufficient flexibility to mimic different crop/pathogen systems. Because we focused on comparative epidemiology rather than on quantitative evaluation, operational conclusions are still out of the scope of the model, and are therefore not presented in this paper. Very few studies have integrated canopy architecture and functioning as a way to control disease [52,53]. Among these studies, the crop-pathogen system is mainly modeled with functional-structural plant framework (FSPM) including 3D representation of the canopy. Using a coarser-grained and less computationally intensive model makes uncertainties (climate variability and change, diverse production systems) easier to analyse within a simulation scheme. Biological complexity is thus easier to integrate and assess [54]. Both models are sensitive to climate (temperature) and show contrasted response. In Mediterranean climate, the disease severity is globally more sensitive variation of model parameters, for the two types of epidemics. In the warmer climate, disease initiations concomitant with a senescent crop (resulting from a hastened crop development) appeared to be more frequent and thus a higher epidemic variance was observed. For the spatial epidemic case, the impact of host ontologic resistance (receptivity) and optimal temperature for pathogen development shifted from none to significant with the change in environmental conditions. This indicates that the model is able to reproduce interesting host\|pathogen\|environment interactions and could be useful to better understand some mechanisms of plant epidemics. Integrative biology can be seen not only as integration between scales but also between the different points of view (agronomists, pathologists and mathematicians). This work also highlighted the role of software simulation platforms as tools to integrate the knowledge coming from these different fields [55]. 1. Oerke E (2006) Crop losses to pests. The Journal of Agricultural Science 144: 31–43. 2. Boote K, Jones J, Mishoe J, Berger R (1983) Coupling pests to crop growth simulators to predict yield reductions. Phytopathology 73: 1581–1587. Sensitivity analysis graphs A and B in figure 3), lower maps resulted from asymmetrical graphs (D and E); right and left maps resulted from a variation in the number of plants considered for local dispersion (4 and 8). doi:10.1371/journal.pone.0049406.g005 November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org 9 Epidemiological Modelling and Crop Architecture Figure 6. Sensitivity analysis. Impact of 10 parameters from table 2 on disease severity for the and bottom, potato late blight. Main effect (circle) and total effect (cross) for each parameters are e contrasted climatic conditions: Mediterranean (black) and oceanic (grey). doi:10.1371/journal.pone.0049406.g006 Figure 6. Sensitivity analysis. Impact of 10 parameters from table 2 on disease severity for the two pathosystems: top, ascochyta blight on pea, and bottom, potato late blight. Main effect (circle) and total effect (cross) for each parameters are estimated with Extended FAST method [41] on two contrasted climatic conditions: Mediterranean (black) and oceanic (grey). doi:10.1371/journal.pone.0049406.g006 Figure 6. Sensitivity analysis. Impact of 10 parameters from table 2 on disease severity for the two pathosystems: top, ascochyta blight on pea, and bottom, potato late blight. Main effect (circle) and total effect (cross) for each parameters are estimated with Extended FAST method [41] on two contrasted climatic conditions: Mediterranean (black) and oceanic (grey). doi:10.1371/journal.pone.0049406.g006 high (or low) model output which can be useful on a prospective point of view. the two pathosystems (ascochyta blight on pea and potato late blight) on two contrasted climatic environments (Oceanic - Rennes, Western France and Mediterranean - Montpellier, South of France). The range of variation for the input parameters was adjusted to keep a biological meaning. We assumed a uniform A global sensitivity analysis of the model was conducted to further apprehend its behavior (cf. figure 6). The impact of 10 parameters on disease severity at the crop scale was assessed for November 2012 \| Volume 7 \| Issue 11 \| e49406 November 2012 \| Volume 7 \| Issue 11 \| e49406 PLOS ONE \| www.plosone.org PLOS ONE \| www.plosone.org 10 Epidemiological Modelling and Crop Architecture 7. Chenu K, Chapman S, Tardieu F, McLean G, Welcker C, et al. (2009) Simulating the yield impacts of organ-level quantitative trait loci associated with 6. Quilot B, Kervella J, Genard M, Lescourret F (2005) Analysing the genetic control of peach fruit quality through an ecophysiological model combined with a QTL approach. Journal of Experimental Botany 56: 3083–3092. 5. Aubertot J, West J, Bousset-Vaslin L, Salam M, Barbetti M, et al. 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https://openalex.org/W3201925748	https://revistas.rcaap.pt/millenium/article/download/24482/18669	English	null	Nurses’ perception of simulation-based basic life support training in clinical practice	DOAJ (DOAJ: Directory of Open Access Journals)	2,021	cc-by	6,441	Corresponding Author Maria João Baptista Rente Rua da Madragoa, Número 98 3880-164 Ovar - Portugal maria.j.b.rente@gmail.com CIÊNCIAS DA VIDA E DA SAÚDE LIFE AND HEALTH SCIENCES CIENCIAS DE LA VIDA Y LA SALUD millenium Millenium, 2(16), 65-72. PERCEÇÃO DOS ENFERMEIROS SOBRE PRÁTICA SIMULADA EM SUPORTE BÁSICO DE VIDA NA PRÁTICA CLÍNICA NURSES’ PERCEPTION OF SIMULATION-BASED BASIC LIFE SUPPORT TRAINING IN CLINICAL PRACTICE PERCEPCIÓN DE LOS ENFERMEROS SOBRE LA PRÁCTICA SIMULADA EN SOPORTE VITAL BÁSICO EN LA PRÁCTICA CLÍNICA en CIÊNCIAS DA VIDA E DA SAÚDE LIFE AND HEALTH SCIENCES CIENCIAS DE LA VIDA Y LA SALUD millenium Millenium, 2(16), 65-72. PERCEÇÃO DOS ENFERMEIROS SOBRE PRÁTICA SIMULADA EM SUPORTE BÁSICO DE VIDA NA PRÁTICA CLÍNICA NURSES’ PERCEPTION OF SIMULATION-BASED BASIC LIFE SUPPORT TRAINING IN CLINICAL PRACTICE PERCEPCIÓN DE LOS ENFERMEROS SOBRE LA PRÁCTICA SIMULADA EN SOPORTE VITAL BÁSICO EN LA PRÁCTICA CLÍNICA en RESUMEN Los participantes fueron 612 enfermeros de diferentes instituciones de salud en Portugal. Los datos se obtuvieron mediante un cuestionario en línea y fueron analizados utilizando el Statistical Package for the Social Sciences (versión 21) con estadística descriptiva e inferencial, según la naturaleza de las variables. Package for the Social Sciences (versión 21) con estadística descriptiva e inferencial, según la naturaleza de las variables. Resultados: El estudio demuestra que los participantes tienen una percepción positiva de las competencias desarrolladas en su práctica clínica, con valores promedio de 4.56±0.71 en la comprensión del algoritmo de soporte vital básico, así como 4.52±0.76 por considerar una excelente contribución a la práctica clínica y 4.51±0.74 en una mejor capacidad para evaluar a la víctima. Conclusión: La práctica simulada es una herramienta essencial en la formación inicial y continua del enfermero. La promoción de un adecuado plan de práctica simulada en soporte vital básico para enfermeros tiene un impacto positivo en la prestación de cuidados dirigidos a las necesidades reales del paciente en parada cardiorrespiratoria Resultados: El estudio demuestra que los participantes tienen una percepción positiva de las competencias desarrolladas en su práctica clínica, con valores promedio de 4.56±0.71 en la comprensión del algoritmo de soporte vital básico, así como 4.52±0.76 por considerar una excelente contribución a la práctica clínica y 4.51±0.74 en una mejor capacidad para evaluar a la víctima. p p y j p p Conclusión: La práctica simulada es una herramienta essencial en la formación inicial y continua del enfermero. La promoción de un adecuado plan de práctica simulada en soporte vital básico para enfermeros tiene un impacto positivo en la prestación de cuidados, dirigidos a las necesidades reales del paciente en parada cardiorrespiratoria. Palabras clave: simulación; reanimación cardiopulmonar; enfermeros ABSTRACT Introduction: Simulation-based basic life support training allows nurses to develop their skills to intervene in an emergency context, solidifying procedures and ensuring safety in practice. etive: Understand nurses’ perception of simulation-based basic life support trainings in clinical practice. Methods: Quantitative, descriptive, and correlational study. Participants were 612 nurses from different health institutions in Portugal. The data was collected through an online questionnaire, and analyzed using the Statistical Package for the Social Sciences (version 21) with descriptive and inferential statistics, according to the nature of the variables. p g Results: The study demonstrates that the participants have a positive perception of the developed skills on their clinical practice, with mean values of 4.56±0.71 for understanding the basic life support algorithm, as well as 4.52±0.76 for considering it an excellent contribution to clinical practice and 4.51±0.74 for improved capacity to assess victims. p p p y Conclusion: Simulation-based training is an essential tool in the initial and continuing education of nurses. The promotion of an adequate simulation-based training plan in basic life support for nurses has a positive impact on care provision, meeting the real needs of the patient in cardiorespiratory arrest. RESUMEN Introducción: La práctica simulada en soporte vital básico permite a los enfermeros desarrollar sus capacidades para intervenir en contexto de emergencia, solidificando procedimientos y garantizando la seguridad en la práctica. en contexto de emergencia, solidificando procedimientos y garantizando la seguridad en la práctica. Objetivo: Comprender la percepción de los enfermeros sobre la práctica simulada en soporte vital básico en la práctica clínica. Métodos: Estudio cuantitativo, descriptivo y correlacional. Los participantes fueron 612 enfermeros de diferentes instituciones de salud en Portugal. Los datos se obtuvieron mediante un cuestionario en línea y fueron analizados utilizando el Statistical Package for the Social Sciences (versión 21) con estadística descriptiva e inferencial, según la naturaleza de las variables. Resultados: El estudio demuestra que los participantes tienen una percepción positiva de las competencias desarrolladas en su práctica clínica, con valores promedio de 4.56±0.71 en la comprensión del algoritmo de soporte vital básico, así como 4.52±0.76 por considerar una excelente contribución a la práctica clínica y 4.51±0.74 en una mejor capacidad para evaluar a la víctima. Conclusión: La práctica simulada es una herramienta essencial en la formación inicial y continua del enfermero. La promoción de un adecuado plan de práctica simulada en soporte vital básico para enfermeros tiene un impacto positivo en la prestación de cuidados, dirigidos a las necesidades reales del paciente en parada cardiorrespiratoria. g p y g g p Objetivo: Comprender la percepción de los enfermeros sobre la práctica simulada en soporte vital básico en la práctica clínica. Objetivo: Comprender la percepción de los enfermeros sobre la práctica simulada en soporte vital básico en la práctica clínica. Métodos: Estudio cuantitativo, descriptivo y correlacional. Los participantes fueron 612 enfermeros de diferentes instituciones Objetivo: Comprender la percepción de los enfermeros sobre la práctica simulada en soporte vital básico en la práctica clínica. Métodos: Estudio cuantitativo, descriptivo y correlacional. Los participantes fueron 612 enfermeros de diferentes instituciones de salud en Portugal. Los datos se obtuvieron mediante un cuestionario en línea y fueron analizados utilizando el Statistical Package for the Social Sciences (versión 21) con estadística descriptiva e inferencial, según la naturaleza de las variables. Resultados: El estudio demuestra que los participantes tienen una percepción positiva de las competencias desarrolladas en su práctica clínica con valores promedio de 4 56±0 71 en la comprensión del algoritmo de soporte vital básico así como 4 52±0 76 Métodos: Estudio cuantitativo, descriptivo y correlacional. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 , , , , , , p , , , ( ) Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 RESUMO Introdução: A prática simulada em suporte básico de vida permite aos enfermeiros desenvolver as suas capacidades para intervir num contexto de emergência, solidificando procedimentos e garantindo a segurança na prática. num contexto de emergência, solidificando procedimentos e garantindo a segurança na prática. Objetivo: Compreender a perceção dos enfermeiros sobre prática simulada em suporte básico de vida na prática clínica. Métodos: Estudo quantitativo, descritivo e correlacional. Os participantes foram 612 enfermeiros de diferentes instituições de saúde em Portugal. Os dados foram adquiridos através de questionário online e analisados com recurso ao Statistical Package for the Social Sciences (versão 21) com estatística descritiva e inferencial, de acordo com a natureza das variáveis. Resultados: O estudo demonstra que os participantes têm uma perceção positiva das competências desenvolvidas na sua prática clínica, com valores médios de 4.56±0.71 na compreensão do algoritmo de suporte básico de vida, bem como 4.52±0.76 por considerarem um excelente contributo para a prática clínica e 4.51±0.74 numa melhor capacidade de avaliação das vítimas. Conclusão: A prática simulada é uma ferramenta essencial na formação inicial e contínua dos enfermeiros. A promoção de um plano de prática simulada adequado em suporte básico de vida para enfermeiros apresenta um impacto positivo na prestação de cuidados indo de encontro às reais necessidades do doente em paragem cardiorrespiratória Objetivo: Compreender a perceção dos enfermeiros sobre prática simulada em suporte básico de vida na prática clínica. Métodos: Estudo quantitativo, descritivo e correlacional. Os participantes foram 612 enfermeiros de diferentes instituições de saúde em Portugal. Os dados foram adquiridos através de questionário online e analisados com recurso ao Statistical Package for the Social Sciences (versão 21) com estatística descritiva e inferencial, de acordo com a natureza das variáveis. Resultados: O estudo demonstra que os participantes têm uma perceção positiva das competências desenvolvidas na sua prática clínica, com valores médios de 4.56±0.71 na compreensão do algoritmo de suporte básico de vida, bem como 4.52±0.76 por considerarem um excelente contributo para a prática clínica e 4.51±0.74 numa melhor capacidade de avaliação das vítimas. ender a perceção dos enfermeiros sobre prática simulada em suporte básico de vida na prática clínica. Objetivo: Compreender a perceção dos enfermeiros sobre prática simulada em suporte básico de vida na prática clínica. Métodos: Estudo quantitativo, descritivo e correlacional. Os participantes foram 612 enfermeiros de diferentes instituições de saúde em Portugal. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). RESUMO Os dados foram adquiridos através de questionário online e analisados com recurso ao Statistical Package for the Social Sciences (versão 21) com estatística descritiva e inferencial, de acordo com a natureza das variáveis. Resultados: O estudo demonstra que os participantes têm uma perceção positiva das competências desenvolvidas na sua prática Métodos: Estudo quantitativo, descritivo e correlacional. Os participantes foram 612 enfermeiros de diferentes instituições de saúde em Portugal. Os dados foram adquiridos através de questionário online e analisados com recurso ao Statistical Package for the Social Sciences (versão 21) com estatística descritiva e inferencial, de acordo com a natureza das variáveis. Resultados: O estudo demonstra que os participantes têm uma perceção positiva das competências desenvolvidas na sua prática clínica, com valores médios de 4.56±0.71 na compreensão do algoritmo de suporte básico de vida, bem como 4.52±0.76 por considerarem um excelente contributo para a prática clínica e 4.51±0.74 numa melhor capacidade de avaliação das vítimas. Resultados: O estudo demonstra que os participantes têm uma perceção positiva das competências desenvolvidas na sua prática clínica, com valores médios de 4.56±0.71 na compreensão do algoritmo de suporte básico de vida, bem como 4.52±0.76 por considerarem um excelente contributo para a prática clínica e 4.51±0.74 numa melhor capacidade de avaliação das vítimas. Conclusão: A prática simulada é uma ferramenta essencial na formação inicial e contínua dos enfermeiros. A promoção de um plano de prática simulada adequado em suporte básico de vida para enfermeiros apresenta um impacto positivo na prestação de Conclusão: A prática simulada é uma ferramenta essencial na formação inicial e contínua dos enfermeiros. A promoção de um plano de prática simulada adequado em suporte básico de vida para enfermeiros apresenta um impacto positivo na prestação de cuidados, indo de encontro às reais necessidades do doente em paragem cardiorrespiratória. Palavras-chave: simulação; reanimação cardiopulmonar; enfermeiros CIÊNCIAS DA VIDA E DA SAÚDE LIFE AND HEALTH SCIENCES CIENCIAS DE LA VIDA Y LA SALUD CIÊNCIAS DA VIDA E DA SAÚDE LIFE AND HEALTH SCIENCES CIENCIAS DE LA VIDA Y LA SALUD 65 Millenium, 2(16), 65-72. en Millenium, 2(16), 65-72. PERCEÇÃO DOS ENFERMEIROS SOBRE PRÁTICA SIMULADA EM SUPORTE BÁSICO DE VIDA NA PRÁTICA CLÍNICA NURSES’ PERCEPTION OF SIMULATION-BASED BASIC LIFE SUPPORT TRAINING IN CLINICAL PRACTICE PERCEPCIÓN DE LOS ENFERMEROS SOBRE LA PRÁCTICA SIMULADA EN SOPORTE VITAL BÁSICO EN LA PRÁCTICA CLÍNICA PERCEÇÃO DOS ENFERMEIROS SOBRE PRÁTICA SIMULADA EM SUPORTE BÁSICO DE VIDA NA PRÁTICA CLÍNICA NURSES’ PERCEPTION OF SIMULATION-BASED BASIC LIFE SUPPORT TRAINING IN CLINICAL PRACTICE PERCEPCIÓN DE LOS ENFERMEROS SOBRE LA PRÁCTICA SIMULADA EN SOPORTE VITAL BÁSICO EN LA PRÁCTICA CLÍNICA Maria Rente1 Mafalda Costa2 https://orcid.org/0000-0001-9706-856X Manuela Pontes3 Fernanda Príncipe4 https://orcid.org/0000-0002-1142-3258 Liliana Mota4 https://orcid.org/0000-0003-3357-7984 1 Local Health Unit of the Alentejo Coast, Santiago do Cacém, Portugal 2 Hospital Center between Douro and Vouga, Santa Maria da Feira, Portugal 3 Portuguese Institute of Oncology, Oporto, Portugal Corresponding Author Maria João Baptista Rente Rua da Madragoa, Número 98 3880-164 Ovar - Portugal maria.j.b.rente@gmail.com RECEIVED: 25th April, 2021 ACCEPTED: 30th July, 2021 RECEIVED: 25th April, 2021 ACCEPTED: 30th July, 2021 66 1. THEORETICAL FRAMEWORK In Portugal, every year, around 10 thousand people are victims of cardiorespiratory arrest Portuguese Cardiology Foundation, n.d.). This epidemiological transition, to global phenomenon, must be considered in nurses’ training programs for victim assistance in emergencies. Team performance may be improved by simulation-based basic life support trainings, executed in a safe environment (Armstrong, Peckler, Pilkinton, McQuade, & Rogan, 2021), in which the participants can learn from their mistakes. The main goals of basic life support are cardiorespiratory arrest prevention and survival improvement after cardiopulmonary resuscitation. Survival chance after sudden cardiorespiratory arrest can be doubled or tripled if cardiopulmonary resuscitation is started quickly. For each minute of delay at the start of cardiopulmonary resuscitation maneuvers, the chances of survival decrease between 7 to 10% (Herbers & Heaser, 2016). This epidemiological transition, to global phenomenon, must be considered in nurses’ training programs for victim assistance in emergencies. Team performance may be improved by simulation-based basic life support trainings, executed in a safe environment (Armstrong, Peckler, Pilkinton, McQuade, & Rogan, 2021), in which the participants can learn from their mistakes. The priorities of cardiopulmonary resuscitation are defined by time and sequence of resuscitator responses, which are the immediate start of chest compressions, immediate defibrillation, and minimization of chest compressions interruptions (Chu & Robilotto, 2018). Resuscitation success and its neurological outcome depend on the efficient and successive implementation of these actions, so simulation-based basic life support training demonstrates positive results in nurses’ clinical practice (Buléon et al., 2016). Thus, even in institutions that have an in-hospital emergency team, all health professionals should have basic life support training to be able to act in situations of peri-arrest and cardiorespiratory arrest until the arrival of a defibrillator and/or specialized team (Vincelette, Quiroz-Martinez, Fortin, & Lavoie, 2018). Nurses are, most of the time, the first to respond to medical emergencies, so they should be confident to act and provide the required care when facing cardiorespiratory arrests; simulation-based basic life support training contributes positively to the development of these skills (Herbers & Heaser, 2016). The basic life support course lasts four hours (European Resuscitation Council, n.d.). Regarding the recertification time, there is no consensus, however the interval of one year between training in basic life support is not enough for your quality execution by nurses (Everett-Thomas et al., 2016). Simulation-based training increases the acquisition/consolidation of skills compared with traditional courses. INTRODUCTION Cardiorespiratory arrest is one of the leading causes of death worldwide, with a global incidence of about 20 thousand people per day (Portuguese Cardiology Foundation, n.d.). Basic life support increases the probability of survival when initiated in the first moments after cardiorespiratory arrest; it comprises two actions: chest compressions and ventilation (Smith, 2017). 67 Recognizing and responding to a cardiorespiratory arrest situation is an event of great stress and anxiety for all nurses, requiring effective performance of cardiopulmonary resuscitation and response prioritization (Sok, Kim, Lee, & Cho, 2020). effective performance of cardiopulmonary resuscitation and response prioritization (Sok, Kim, Lee, & Cho, 2020). Sudden events are present in nurses’ daily life and these professionals must be ready to act and implement life-saving interventions. Simulation-based basic life support training provides an effective and safe environment for nurses to learn and practice the clinical skills necessary in emergencies (Morton, 2017). Sudden events are present in nurses’ daily life and these professionals must be ready to act and implement life-saving interventions. Simulation-based basic life support training provides an effective and safe environment for nurses to learn and practice the clinical skills necessary in emergencies (Morton, 2017). Nurses are able to develop their capabilities to intervene in emergencies through simulation-based basic life support, solidifying procedures and ensuring practice safety. The simulation effects on nursing practice include greater knowledge of clinical competence, communication and clinical decision making skills, problem solving, and critical thinking (Kim & Choi, 2019). Nurses are able to develop their capabilities to intervene in emergencies through simulation-based basic life support, solidifying procedures and ensuring practice safety. The simulation effects on nursing practice include greater knowledge of clinical competence, communication and clinical decision making skills, problem solving, and critical thinking (Kim & Choi, 2019). This investigation study aims to understand nurses’ perception of simulation-based basic life support training on the clinical i f h i i i i kill i i i competence, communication and clinical decision making skills, problem solving, and critical thinking (Kim & Choi, 2019). This investigation study aims to understand nurses’ perception of simulation-based basic life support training on the clinical practice of nurses, to emphasize its importance in skills acquisition. This investigation study aims to understand nurses’ perception of simulation-based basic life support training on the clinical practice of nurses, to emphasize its importance in skills acquisition. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 m 67 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 2. METHODS This is a quantitative, descriptive, and correlational study. m16 identification and understanding of the problems in implementing techniques and the proper training of professionals in emergencies. Continuous training in basic life support, carried out through simulation-debriefing, is an effective way to acquire adequate and long-lasting knowledge. The achieved satisfaction level is high since this learning method corresponds to professionals’ expectations and it is similar to real clinical practice (Fraga-Sampedro, 2018). identification and understanding of the problems in implementing techniques and the proper training of professionals in emergencies. Continuous training in basic life support, carried out through simulation-debriefing, is an effective way to acquire adequate and long-lasting knowledge. The achieved satisfaction level is high since this learning method corresponds to professionals’ expectations and it is similar to real clinical practice (Fraga-Sampedro, 2018). The debriefing session after the practice ensures that professionals learn from the experience and value their participation in basic lif (H b & H 2016) p p p ( g p ) The debriefing session after the practice ensures that professionals learn from the experience and value their participation in basic life support (Herbers & Heaser, 2016). 2.3 Statiscal analysis For statistical analysis, descriptive and inferential statistics were used with the statistical Package for the Social Sciences Program version 21. All questions were answered by the participants. Data normality was verified with the Shapiro-Wilk test. The perception of simulation-based skills of basic life support were correlated with the categorical variables using the Pearson correlation coefficient. The significance level was set at p < 0.05. 2.2 Data collection instruments and procedures Data collection was carried out using the online questionnaire, available during June 2020. ng the online questionnaire, available during June 20 The data collection instrument consists of two parts: Part 1 — sociodemographic characterization; and Part 2 — perception of simulation-based skills of basic life support. For socio-demographic characterization, the following characteristics were used: sex, age, educational qualifications, job category, area of specialization, years of experience, training in basic life support, the date of the last training in basic life support, the last training in basic life support attended–whether it was a formal training course in basic life support and if it was simulation-based. The second part of the questionnaire consists of 13 items, operationalized on a Likert-like scale that varies between semantic fields (1 — totally disagree; 2 — disagree; 3 — neither disagree / nor agree; 4 – agree; 5 — totally agree). The internal consistency value (Cronbach's Alpha) was 0.94. Participation in the study was voluntary with guaranteed anonymity; the participants could withdraw from the study without any prejudice. The study was approved by the Ethics Committee (Protocol 08/2020). 2.1 Sample The target population was Portuguese nurses; the sample was non-probabilistic and by convenience. The participants were 612 nurses who develop their practice in clinical health care contexts and accepted to participate in the study. Nurses who exercised their practice exclusively in the field of teaching were excluded from the study. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). 1. THEORETICAL FRAMEWORK Simulation-based training can be of low, medium, or high fidelity. Low-fidelity simulation is developed using role playing/case studies; medium- fidelity simulation involves realistic scenarios with simple mannequins; high-fidelity simulation uses realistic scenarios in situ or within simulation centers, using computer-controlled mannequins (Chu & Robilotto, 2018). Compared with traditional trainings, high-fidelity simulation allows professionals to practice and apply skills in basic life support situations and to learn in a safe environment. Thus, frequent simulations improve basic life support skills (Halm & Crespo, 2018). Simulation-based training in situ are a quick and efficient way of applying theoretical knowledge and developing muscle memory; it is also a great teamwork opportunity. Moreover, basic life support simulations significantly improve response time and increase the confidence of professionals (Herbers & Heaser, 2016). The adequate performance of a basic life support team relies on technical and non-technical skills; non-technical skills are leadership, teamwork, proper task management, and interpersonal communication. Highly effective teams are associated with coordinated functions of leadership and clarity of responsibilities when dealing with cardiorespiratory arrests (Armstrong et al., 2021). Teamwork is crucial and its development in simulation-based training is necessary for the effective evaluation of patients, due to increased specialization and complexity of care, risks associated with treatment options, and the need to ensure patient safety (Armstrong et al., 2021). Effective resuscitation requires a coordinated response from several professionals. The quality of teamwork can affect results. Identifying an efficient procedure for in-hospital response to emergencies involves at least four teamwork functions: shared mental models, clarity of roles, communication, and workload management (Armstrong, et al., 2021). Basic life support simulation with debriefing has positive effects on the way nurses control emotions, as well as performance and cognitive skills; moreover, it is an excellent pedagogical tool because due to being practical and objective, allowing the 68 m16 3. RESULTS Regarding the sociodemographic characteristics of the 612 participants (Table 1), we found that they were on average 39.9±9.8 years old — the youngest was 22 and the oldest was 64 years old. Most of them were female (79.7%) The overall qualification degree was licensed; Medical-surgical Nursing was the most frequent specialization. They had an average of 16.3±10.1 years of professional experience, ranging from 0 to 44 years. Most participants of the study work in Urgency and Emergency sectors. k in Urgency and Emergency sectors. Most participants of the study work in Urgency and Emergency sectors. Table 1 – Sociodemographic characterization of nurses. Variable N % Qualifications Bachelor 2 0.3 Licensed 479 78.3 Master 127 20.8 PhD 4 0.7 Professional Category Nurse 373 60.9 Specialist Nurse 231 37.7 Nurse Manager 8 1.3 Nursing Specialties Medical-Surgical 109 17.8 Rehabilitation nursing 44 7.2 Community health nursing 40 6.5 Psychiatric nursing 15 2.5 Pediatric nursing 31 5.1 Obstetrical nursing 22 3.6 Table 1 – Sociodemographic characterization of nurses. 69 m16 Most nurses claim to have basic life support training (98.2%), and on average the last training occurred 3.4±3.48 years ago (ranging from 0 to 30 years ago). Regarding the last basic life support training, 90.8% of the nurses reported that it was a formal course and 97.7% of them were simulation-based. Regarding the acquired skills in simulation-based training, the participants presented higher mean values of understanding the basic life support algorithm and considering it an excellent contribution to clinical practice — mainly in better evaluating the victim (Table 2). Table 2 – Descriptive statistics of simulation-based skills. Table 2 – Descriptive statistics of simulation-based skills. Table 2 – Descriptive statistics of simulation-based skills. Simulation-based skills Positive feedback after simulation-based training Realism of basic life support activities Realism of simulation-based training The lower mean value is related to the simulation realism of the stress level, which are higher in clinical practice than in basic life support trainings. The Pearson correlation (r) verified that the different variables present a significant and negative correlation with age and professional experience (Table 3). The lower mean value is related to the simulation realism of the stress level, which are higher in clinical practice than in basic life support trainings. Th P l ti ( ) ifi d th t th diff t i bl t i ifi t d ti l ti ith d The lower mean value is related to the simulation realism of the stress level, which are higher in clinical p support trainings. The Pearson correlation (r) verified that the different variables present a significant and negative co professional experience (Table 3). able 3 – Correlation matrix of the variables with age and professional experience. Table 3 – Correlation matrix of the variables with age and professional experience. Simulation-based skills Age Professional experience Contribution of regular basic life support skills training on my clinical practice r -.183 -.159 Understanding of the basic life support algorithm r -.200 -.175 Knowledge of the European Resuscitation Council guidelines r -.197 -.177 Capacity of assessing victims r -.130 -.110 Improvement of my technical and non-technical skills r -.165 -.143 Teamwork r -.165 -.153 Self-confidence r -.174 -.163 Positive feedback after simulation-based training r -.216 -.196 Realism of basic life support activities r -.092* -.075 Realism of clinical cases r -.139 -.121 Realism of simulation-based training r -.135 -.119 Realism of stress level r .055 .043 Relevance of simulation-based basic life support practical courses r -.119 -.121 * Significant correlation for p = 0.05. ** Significant correlation for p < 0.001. Positive feedback after simulation-based training Realism of basic life support activities Realism of simulation-based training ealism of stress level Relevance of simulation-based basic life support practical courses * Significant correlation for p = 0.05. ** Significant correlation for p < 0.001. We found that as the age and professional experience of the participant’s increases, they recognize less impact of simulation- based basic life support trainings in their clinical practice in the different variables evaluated. Age and experience did not correlate with realism of stress level in simulation-based training. 3. RESULTS Simulation-based skills Minimum Maximum Mean Standard deviation Contribution of regular basic life support skills training on my clinical practice 1 5 4.52 0.76 Understanding of the basic life support algorithm 1 5 4.56 0.71 Knowledge of the European Resuscitation Council guidelines 1 5 4.23 0.96 Capacity of assessing victims 1 5 4.51 0.74 Improvement of my technical and non-technical skills 1 5 4.48 0.76 Teamwork 1 5 4.27 0.85 Self-confidence 1 5 4.11 0.96 Positive feedback after simulation-based training 1 5 4.31 0.82 Realism of basic life support activities 1 5 4.06 0.91 Realism of clinical cases 1 5 4.08 0.89 Realism of simulation-based training 1 5 4.03 0.92 Realism of stress level 1 5 3.04 1.07 Relevance of simulation-based basic life support practical courses 1 5 3.97 0.97 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 m 69 Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. 69 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). N ’ i f i l i b d b i lif i i i li i l i Mill i 2(16) 65 72 69 m16 We found with the assessment of the data that simulation-based training of basic life support promotes better capacity of assessing victims, with high mean values that are corroborated by the evidence (Halm & Crespo, 2018) and improves technical and non-technical skills (Kim & Choi, 2019; Więch et al., 2019). ( ę ) We noticed from the analysis of the collected data that simulation-based training encourages teamwork, as evidenced in the studies by Colquitt et al. (2019) and Halm & Crespo (2018), which increases nurses’ self-confidence in non-technical skills, such as interpersonal communication (Kim & Choi, 2019; Morton, 2017). In the data analysis, we can also verify that nurses gave positive feedback after receiving simulation-based basic life support training, as evidenced by Kim, & Choy (2019), since the activities and clinical cases of simulation-based basic life support were realistic, as in the studies by Halm & Crespo (2018) and Whitmore et al., (2019). g, y , y ( ), istic, as in the studies by Halm & Crespo (2018) and Whitmore et al., (2019). Although nurses’ answers do not present high mean levels regarding the level of stress, the evidence argues that simulation-based training decreases stress levels experienced by nurses in real contexts (Sok et al., 2020). From the data analysis, it appears that the simulation-based courses have a greater impact on clinical practice of nurses than conventional trainings, as corroborated by other studies (Halm & Crespo, 2018; Kim & Choi, 2019). From the data analysis, it appears that the simulation-based courses have a greater impact on clinical practice of nurses than conventional trainings, as corroborated by other studies (Halm & Crespo, 2018; Kim & Choi, 2019). Since cardiorespiratory arrest is a sudden event that requires priority response and cardiopulmonary resuscitation, the simulation- based basic life support training allows nurses to develop skills to intervene in emergencies, with a high analyzed mean value. The acquired and consolidated skills contribute not only to teamwork, but also help team members to focus on the care priorities during cardiopulmonary resuscitation and situation management (Colquitt et al., 2019; Halm & Crespo, 2018; Whitmore et al., 2019). Regarding basic life support, simulation-based training increases performance effectiveness and develops critical thinking and decision-making skills, which promote confidence in clinical nursing practice, attesting the mean values obtained, increasing nurses’ satisfaction and competence (Halm & Crespo, 2018; Kim & Choi, 2019; Whitmore et al., 2019). m16 However, the statistical analysis showed that as the age of participants and professional experience increase, professionals recognize less impact of simulation-based trainings on their skills. So, the simulation-based models of basic life support training should be rethought to better develop skills throughout the entire professional cycle of nurses. Basic life support training with simulation improves nurses’ competence and performance times in resuscitation situations, proving with evidence, which promotes the reduction of complications during provision of care, as well as the difficulties experienced in sudden events (Fraga-Sampedro, 2018; Whitmore et al., 2019). Therefore, it is important to include Good Training Guides when planning simulation-based courses. Nurses reported that the acquisition/consolidation of basic life support skills was significantly positive in simulation-based trainings, with high mean values, compared with conventional trainings. Frequent simulation-based training improves skills and prevents their degradation (Halm & Crespo, 2018). High fidelity simulation-based training with advanced software and realistic scenarios provides clinical realism to basic life support formation, which promotes the understanding and involvement of nurses (Everett-Thomas et al., 2016; Halm & Crespo, 2018; Whitmore et al., 2019). High fidelity simulation-based training with advanced software and realistic scenarios provides clinical realism to basic life support formation, which promotes the understanding and involvement of nurses (Everett-Thomas et al., 2016; Halm & Crespo, 2018; Whitmore et al., 2019). , ) Basic life support simulations do not replace real experience, but it is of higher fidelity compared with other training methods (Fraga-Sampedro, 2018). Basic life support simulations do not replace real experience, but it is of higher fidelity compared with other training methods (Fraga-Sampedro, 2018). Analyzing the knowledge and skills acquired by nurses in simulation-based basic life support trainings, we found that they positively perceive the benefit for clinical practice. Analyzing the knowledge and skills acquired by nurses in simulation-based basic life support trainings, we found that they positively perceive the benefit for clinical practice. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). 4. DISCUSSION Data analysis of knowledge and skills acquired in simulation-based basic life support training show that recurring training contributes to the clinical practice of nurses (Colquitt, Walker, & Haney, 2019; Whitmore et al., 2019). m16 70 70 REFERENCES Armstrong, P., Peckler, B., Pilkinton, C. J., McQuade, D., & Rogan, A. (2021). Effect of simulation training on nurse leadership in a shared leadership model for cardiopulmonary resuscitation in the emergency department. Emergency Medicine Australasia, 33(2), 255–261. doi: 10.1111/1742-6723.13605. Buléon, C., Delaunay, J., Parienti, J.-J., Halbout, L., Arrot, X., Gérard, J.-L., & Hanouz, J.-L. (2016). Impact of a feedback device on chest compression quality during extended manikin CPR: a randomized crossover study. The American Journal of Emergency Medicine, 34(9), 1754–1760. doi: 10.1016/j.ajem.2016.05.077. Chu, R., & Robilotto, T. (2018). Mock code training to enhance CPR skills. Nursing Made Incredibly Easy, 16(2), 11–15. doi: 10.1097/01.NME.0000529957.11904.8d. Colquitt, J. D., Jr, Walker, A. B., & Haney, N. S. (2019). Applying the Pit Crew Resuscitation Model to the Inpatient Care Setting. Journal for Nurses in Professional Development, 35(1), E1–E7. doi: 10.1097/NND.0000000000000495. tation Council. (n.d.). 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Implementing an in Situ Mock Code Quality Improvement Program. American Journal of Critical Care: An Official Publication, American Association of Critical-Care Nurses, 25(5), 393–399. doi: 10.4037/ajcc2016583. Kim, S.-O., & Choi, Y.-J. (2019). Nursing competency and educational needs for clinical practice of Korean nurses. Nurse Education in Practice, 34, 43–47. doi: 10.1016/j.nepr.2018.11.002. Morton, S. B. (2017). Impact of High-Fidelity Simulation Training on Medicalsurgicalnurses’ Self-Confidence & Mock Code Blue Performance: A Pilot Study (Doctoral dissertation). Obtained from https://ninercommons.uncc.edu/islandora/object/etd%3A1022/datastream/PDF/download/citation.pdf. Portuguese Cardiology Foundation (n.d.). Statistic data. http://www.fpcardiologia.pt/atividades/projeto-salva-vidas/dados- estatisticos/. Smith, L. R. (2017). Improving Cardiopulmonary Resuscitation Skills Using Unit Based Simulation (Doctoral project). Obtained from https://digitalcommons.gardner-webb.edu/cgi/viewcontent.cgi?article=1296&context=nursing_etd. Sok, S. R., Kim, J. A., Lee, Y., & Cho, Y. (2020). Effects of a Simulation-Based CPR Training Program on Knowledge, Performance, and Stress in Clinical Nurses. Journal of Continuing Education in Nursing, 51(5), 225–232. doi: 10.3928/00220124- 20200415-07. CONCLUSION Simulation-based learning is an essential tool in the initial and continuous training of nurses, crucial to maintain and train basic life support skills. The training situations should be as realistic as possible, in a pedagogical and safe environment. The representation of clinical reality and stress environments are fundamental in the context of cardiorespiratory arrest. In situations of cardiopulmonary resuscitation, the simulated training appears as one of the most promising pedagogical options, since it overcomes the difficulties in adequately training nurses to act under stress in sudden events. It is an effective improvement in safety, efficacy, and nursing care in basic life support to critical patients. To ensure the efficiency of training, it is necessary to develop clinical cases that can transport the trainee to a reality similar to sudden events, whether in incidents, difficulties, or stress. Simulation-based trainings should be implemented unequivocally, whether in initial or continuous trainings, because they provide active and systematic learning experiences, enhancing the development of technical and non-technical skills, such as interpersonal communication and teamwork. Allowing nurses to demonstrate their knowledge, skills, attitudes, and behaviors in a safe environment. Simulations create an integrated vision that facilitates the balance between the adequate provision of care, the fulfillment of basic life support, and the correct execution of the maneuvers. 71 Adequate simulation-based training in basic life support for nurses has a positive impact on the provision of care that meets the real needs of patients suffering from cardiorespiratory arrest. Nurses should rely on these practices to answer questions regarding cardiorespiratory arrest. In nursing, research is indispensable to allow the acquisition of skills that contribute, not only to improve people’s quality of life, but also to recognize and appreciate the nursing profession. In future studies, it is important to analyze the impact of planning of basic life support trainings, using good training guides to develop nurses’ clinical skills. ACKNOWLEDGEMENTS We thank all participants who dedicated themselves to this study and the translators for their work. 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R., VanDyck, T., Hebert, C., … Neumar, R. W. (2019). Simulation training enables emergency medicine providers to rapidly and safely initiate extracorporeal cardiopulmonary resuscitation (ECPR) in a simulated cardiac arrest scenario. Resuscitation, 138, 68–73. doi: 10.1016/j.resuscitation.2019.03.002. Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). Nurses’ perception of simulation-based basic life support training in clinical practice. Millenium, 2(16), 65-72. DOI: https://doi.org/10.29352/mill0216.24482 Rente, M., Costa, M., Pontes, M., Príncipe, F., & Mota, L. (2021). REFERENCES Vincelette, C., Quiroz-Martinez, H., Fortin, O., & Lavoie, S. (2018). Timely Recognition of Ventricular Fibrillation and Initiation of Cardiopulmonary Resuscitation by Intensive Care Unit Nurses: A High-Fidelity Simulation Observational Study. Clinical Simulation in Nursing, 23, 1–9. doi: 10.1016/j.ecns.2018.07.005. 72 m16 Whitmore, S. P., Gunnerson, K. J., Haft, J. W., Lynch, W. R., VanDyck, T., Hebert, C., … Neumar, R. W. (2019). Simulation training enables emergency medicine providers to rapidly and safely initiate extracorporeal cardiopulmonary resuscitation (ECPR) in a simulated cardiac arrest scenario. Resuscitation, 138, 68–73. doi: 10.1016/j.resuscitation.2019.03.002. Więch, P., Sałacińska, I., Muster, M., Bazaliński, D., Kucaba, G., Fąfara, A., … Januszewicz, P. (2019). Use of Selected Telemedicine Tools in Monitoring Quality of In-Hospital Cardiopulmonary Resuscitation: A Prospective Observational Pilot Simulation Study. Medical Science Monitor: International Medical Journal of Experimental and Clinical Research, 25, 2520–2526. doi: 10.12659/MSM.913191.
https://openalex.org/W3159366615	https://www.mdpi.com/2073-4433/12/5/601/pdf?version=1620372256	English	null	Characteristics of Particle Size Distributions of Falling Volcanic Ash Measured by Optical Disdrometers at the Sakurajima Volcano, Japan	Atmosphere	2,021	cc-by	13,978	Keywords: disdrometer; eruption cloud; fall velocity; gamma function; particle size distribution; quantitative ash fall estimation; Sakurajima; volcanic ash; weather radar Academic Editors: Meelis Zidikheri and Lars Hoffman Received: 5 February 2021 Accepted: 21 April 2021 Published: 6 May 2021 Received: 5 February 2021 Accepted: 21 April 2021 Published: 6 May 2021 Received: 5 February 2021 Accepted: 21 April 2021 Published: 6 May 2021 atmosphere atmosphere atmosphere atmosphere Masayuki Maki 1,, Ren Takaoka 2 and Masato Iguchi 3 1 Research and Education Center for Natural Hazards, Kagoshima University, Kagoshima 890-0065, Japan 2 Graduate School of Science and Engineering, Kagoshima University, Kagoshima 890-0065, Japan; k6636104@kadai.jp 3 Disaster Prevention Research Institute, Kyoto University, Kyoto 611-0011, Japan; iguchi.masato.8m@kyoto-u.ac.jp Correspondence: maki@gm.kagoshima-u.ac.jp 1 Research and Education Center for Natural Hazards, Kagoshima University, Kagoshima 890 1 Research and Education Center for Natural Hazards, Kagoshima University, Kagoshima 890-0065, Japan 2 Graduate School of Science and Engineering, Kagoshima University, Kagoshima 890-0065, Japan; k6636104@kadai.jp 3 Disaster Prevention Research Institute, Kyoto University, Kyoto 611-0011, Japan; iguchi.masato.8m@kyoto-u.ac.jp * Correspondence: maki@gm.kagoshima-u.ac.jp 1 Research and Education Center for Natural Hazards, Kagoshima University, Kagoshima 890-0065, Japan 2 Graduate School of Science and Engineering, Kagoshima University, Kagoshima 890-0065, Japan; k6636104@kadai.jp 3 jp 3 Disaster Prevention Research Institute, Kyoto University, Kyoto 611-0011, Japan; iguchi.masato.8m@kyoto-u.ac.jp Abstract: In the present study, we analyzed the particle size distribution (PSD) of falling volcanic ash particles measured using optical disdrometers during six explosive eruptions of the Sakurajima volcano in Kagoshima Prefecture, Japan. Assuming the gamma PSD model, which is commonly used in radar meteorology, we examined the relationships between each of the gamma PSD parameters (the intercept parameter, the slope parameter, and the shape parameter) calculated by the complete moment method. It was shown that there were good correlations between each of the gamma PSD parameters, which might be one of the characteristics of falling volcanic ash particles. We found from the normalized gamma PSD analysis that the normalized intercept parameter and mass-weighted mean diameter are suitable for estimating the ash fall rate. We also derived empirical power law relationships between pairs of integrated PSD parameters: the ash fall rate, the volcanic ash mass concentration, the reﬂectivity factor, and the total number of ash particles per unit volume. The results of the present study provide essential information for studying microphysical processes in volcanic ash clouds, developing a method for quantitative ash fall estimation using weather radar, and improving ash transport and sedimentation models. Citation: Maki, M.; Takaoka, R.; Iguchi, M. Characteristics of Particle Size Distributions of Falling Volcanic Ash Measured by Optical Disdrometers at the Sakurajima Volcano, Japan. Atmosphere 2021, 12, 601. https://doi.org/10.3390/ atmos12050601 Article Characteristics of Particle Size Distributions of Falling Volcanic Ash Measured by Optical Disdrometers at the Sakurajima Volcano, Japan Masayuki Maki 1,*, Ren Takaoka 2 and Masato Iguchi 3 1. Introduction The physical properties and chemical components of ash particles emitted by explo- sive volcanic eruptions represent useful basic data for investigating the mode, scale, and mechanisms of eruptions. Previous studies have investigated the generation mechanism of individual eruptions [1,2] and the fragmentation process during explosive eruptions by estimating the total grain size distribution following sediment surveys [3–5]. Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional afﬁl- iations. In recent years, volcanic sediment data analysis has facilitated many types of research, such as the detection, tracking, and prediction of ash fall to prevent volcanic disasters [6–8], estimation of the content fraction of particles smaller than 63 µm (which affect aircraft operation) from total grain size distribution [7], and investigation of variation in tephra features with distance from the volcano crater using the 100-year eruption records of volcanoes along the western coast of North America [8]. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). Although investigating ground sediments deposited following volcanic eruptions is an important method in volcanology research, and for volcanic disaster prevention, sediment data acquired on the ground are inevitably time-integrated, they are usually accumulated hours or days after an eruption. Large historical eruptions have resulted in centuries of accumulated sediment. Conventional geological methods for investigating https://www.mdpi.com/journal/atmosphere Atmosphere 2021, 12, 601. https://doi.org/10.3390/atmos12050601 Atmosphere 2021, 12, 601 2 of 24 sediments are limited in that the properties and sedimentary environment of volcanic ash particles change depending on weather conditions, such as wind and rain, during and after sedimentation [9], such that sediment data are often unsuitable for remote sensing and numerical forecasting of ash fall distribution and transportation, which change from moment to moment. Recent studies have attempted to clarify the physical properties of ash particles at high temporal resolution using meteorological optical disdrometers, which were developed to measure the physical characteristics of precipitation particles [10–12] including particle size distribution (PSD), shape, falling velocity, and canting. Optical disdrometers have also been used to study microphysics in precipitation clouds, to develop quantitative precipitation estimates, and to provide ground truth data for satellite remote sensing of precipitation. Because the optical disdrometer is a non-contact sensor, it can measure falling volcanic ash particles without altering local environmental conditions. 1. Introduction A certain type of optical disdrometer can acquire the shape of a falling single particle using its fast scanning frequency, which is typically a few thousands per second. q y yp y p Free-fall experiments conducted in a large experimental facility measured the falling velocity, shape, and canting angles of ash particles using a two-dimensional video disdrom- eter (2DVD) [10], which was developed to observe precipitation particles [13,14]. Another type of optical disdrometer, Parsivel2 [15,16], was installed on Sakurajima, a volcano in Kagoshima Prefecture, Japan to automatically measure falling ash particles [11]. This in- strument measured temporal changes in the PSD and fall velocity of tephra from a total of 76 eruptions of Sakurajima over a 2-year period. An analysis of these records showed that temporal changes in the Parsivel2 PSD data were synchronized with the crustal movements associated with eruptions. Based on the success mentioned above, automated measure- ments of ash particles are ongoing with 13 Parsivel2 instruments that have been arranged in all directions centering on the Minami-dake summit of Sakurajima [12]. The goal of the present study was to ﬁnd characteristics of volcanic ash PSD. One of the unique features of the present study is an analysis of PSD data measured by disdrometers every 1 min. The results obtained will be useful for the radar monitoring of ash falls and improvement of volcanic ash transport and diffusion models. The structure of the present paper is as follows. In Section 2, we describe the princi- ples by which ash particles are measured by Parsivel2, the data analysis, and processing methods. In Section 3, we describe the functional forms and integrated parameters of ash particle PSD, which provide a theoretical basis for quantitative ash fall estimation and analysis of volcanic ash cloud microphysics. Our results are described and discussed in Sections 4 and 5, respectively. The phenomenon of fragmentation, which is an important eruption mechanism based on the physical and chemical characteristics of volcanic ash particles, is beyond the scope of this study, and will not be discussed. 2.1. Parsivel2 The particle diameter is estimated from ∆V, and fall velocity is estimated from ∆t. Table 1. Main speciﬁcations of the Parsivel2 disdrometer, according to the instruction manual (OTT, 2011). Category Subcategory Speciﬁcation Sensor Type Laser diode Wavelength 650 nm Output rating 0.2 mW Laser class 1 (IEC/EN 60825-1: 2014) Sampling Light strip (W × D) 30 × 1 mm (54 cm2) Area (W × D) 180 × 30 mm (54 cm2) Interval 10 s to 60 min Output Particle size 32 classes: 0.2, . . . , 8 mm (liquid particles) 32 classes: 0.2, . . . , 25 mm (solid particles) Fall velocity 32 classes: 0.2, . . . , 20 m s−1 Precipitation intensity (accuracy) 0.001, . . . , 1200 mm h−1 (±5% for liquid, ±20% for solid) Precipitation type 8 types (drizzle, drizzle/rain, rain, snow, mixed rain/snow, snow grains, sleet, hail) Radar reﬂectivity factor −9.999, . . . , 99.999 dBZ ± 20% Kinetic energy 0, . . . , 999.999 J/(m2 h) Visibility in precipitation 0, . . . , 20,000 m Figure 1. Schematic diagram illustrating ash particle diameter and fall velocity measurements by Parsivel2 (Löfﬂer-Mang and Joss, 2000). As a particle passes through the measurement laser beam sheet, voltage reduction ∆V is generated during time interval ∆t at the light-receiving device. The particle diameter is estimated from ∆V, and fall velocity is estimated from ∆t. The PSD estimation error for Parsivel has been compared with that for other in- Table 1. Main speciﬁcations of the Parsivel2 disdrometer, according to the instruction manual (OTT, 2011). Figure 1. Schematic diagram illustrating ash particle diameter and fall velocity measurements by Parsivel2 (Löfﬂer-Mang and Joss, 2000). As a particle passes through the measurement laser beam sheet, voltage reduction ∆V is generated during time interval ∆t at the light-receiving device. The particle diameter is estimated from ∆V, and fall velocity is estimated from ∆t. The PSD estimation error for Parsivel has been compared with that for other in- struments [15,16,18–20]. The accuracy of the ﬁrst-generation Parsivel and Parsivel2 was evaluated by comparing measured data with those from an impact disdrometer observing raindrops [16,21]. The results showed that Parsivel2 has improved the accuracy of rainfall and PSD measurements of particles 0.5 and 4 mm in size. The accuracy of Parsivel2 is acceptable for estimating the fall velocity of particles < 1 mm in diameter. 2.1. Parsivel2 In this study, we used the Parsivel2 (PARticle SIze and VELocity) optical disdrometer; its main speciﬁcations are listed in Table 1 [17]. Parsivel2 measures falling particles as follows. A ﬂat, horizontal laser measuring surface (180 mm × 30 mm × 1 mm) is formed between the light-emitting and light-receiving devices. The diameter (0.2–25 mm) and fall speed (up to 20 ms−1) of the particles are calculated in real time based on the voltage drop and duration as falling particles pass through the laser measurement surface (Figure 1). In these calculations, it was assumed that particles < 1 mm in diameter are spherical, the axial ratio of particles 1–5 mm in diameter varies linearly from 1 to 0.7, and the axial ratio of particles > 5 mm in diameter is 0.7. These assumptions are based on the original function of Parsivel, which was designed to measure raindrops, and may lead to errors in measurements of volcanic ash particles. At present, it is difﬁcult to estimate these errors because the relationship between the axis ratio and diameter of volcanic ash particles is unclear. Atmosphere 2021, 12, 601 3 of 24 Table 1. Main speciﬁcations of the Parsivel2 disdrometer, according to the instruction manual (OTT, 2011). Category Subcategory Speciﬁcation Sensor Type Laser diode Wavelength 650 nm Output rating 0.2 mW Laser class 1 (IEC/EN 60825-1: 2014) Sampling Light strip (W × D) 30 × 1 mm (54 cm2) Area (W × D) 180 × 30 mm (54 cm2) Interval 10 s to 60 min Output Particle size 32 classes: 0.2, . . . , 8 mm (liquid particles) 32 classes: 0.2, . . . , 25 mm (solid particles) Fall velocity 32 classes: 0.2, . . . , 20 m s−1 Precipitation intensity (accuracy) 0.001, . . . , 1200 mm h−1 (±5% for liquid, ±20% for solid) Precipitation type 8 types (drizzle, drizzle/rain, rain, snow, mixed rain/snow, snow grains, sleet, hail) Radar reﬂectivity factor −9.999, . . . , 99.999 dBZ ± 20% Kinetic energy 0, . . . , 999.999 J/(m2 h) Visibility in precipitation 0, . . . , 20,000 m Figure 1. Schematic diagram illustrating ash particle diameter and fall velocity measurements by Parsivel2 (Löfﬂer-Mang and Joss, 2000). As a particle passes through the measurement laser beam sheet, voltage reduction ∆V is generated during time interval ∆t at the light-receiving device. 2.1. Parsivel2 Parsivel2 was found to underestimate fall velocity near 1 m s−1, but this trend decreased as particle size increased. 2.2. Data Collection The ash particle data analyzed in this study were collected by the Parsivel2 network (Figure 2), which was installed on Sakurajima and operated by the Disaster Prevention Re- search Institute of Kyoto University. We extracted PSD data, including eruption onset time Atmosphere 2021, 12, 601 4 of 24 and cloud top height, from the Parsivel2 database for six volcanic eruptions that occurred in 2018 (Table 2), according to reports of Sakurajima volcanic eruptions published by the Kagoshima Local Meteorological Observatory of the Japan Meteorological Agency [22,23]. and cloud top height, from the Parsivel2 database for six volcanic eruptions that occurred in 2018 (Table 2), according to reports of Sakurajima volcanic eruptions published by the Kagoshima Local Meteorological Observatory of the Japan Meteorological Agency [22,23]. Figure 2. The network of 13 Parsivel2 instruments installed on Sakurajima to measure volcanic ash particle size distribution (PSD). Triangle indicates the southern summit (Minami-dake); square indicates the position of X-band multi-parameter radar (XMP). Figure 2. The network of 13 Parsivel2 instruments installed on Sakurajima to measure volcanic ash particle size distribution (PSD). Triangle indicates the southern summit (Minami-dake); square indicates the position of X-band multi-parameter radar (XMP). Table 2. List of Sakurajima volcanic eruptions analyzed in the present study. Table 2. List of Sakurajima volcanic eruptions analyzed in the present study. Case Eruption Onset (LST) Cloud Top Height (m) Movement Disdrometer Code 1 08:08, 15 May 2018 2500 Top NABE 2 19:48, 22 May 2018 Unknown Unknown HIKP, HART 3 08:01, 30 May 2018 2500 Eastward KURP 4 11:35, 10 June 2018 3500 Top HART 5 07:19, 16 June 2018 4700 Westward SBTT 6 15:38, 16 July 2018 4600 Westward AKAM To conﬁrm that an eruption cloud had passed over a Parsivel2 site, we used X-band multi-parameter radar (XMP), which was installed approximately 11 km from Sakurajima’s Minami-dake summit. XMP is operated by the Ministry of Land, Infrastructure and Trans- port to monitor debris ﬂows on Sakurajima; although it is used to observe precipitation, it has also been shown to capture ash particles falling from eruption clouds [24]. Eruption cases were selected for analysis as follows. First, eruptions in which the eruption cloud top height was >2000 m above the vent were extracted. 2.2. Data Collection Then, XMP data for the extracted eruptions were downloaded from the Extended Rainfall Indicator Network (XRAIN) data download system [25] to draw time-cumulative plan position indicator (PPI) images of reﬂectivity at 2 elevation angles, of 1.7◦and 6◦. Finally, eruptions in which the installation point of Parsivel2 was within the cumulative reﬂectivity distribution area were selected for analysis. Atmosphere 2021, 12, 601 5 of 24 2.3. Data Processing Various physical quantities of particles falling through the atmosphere were calculated from PSD data measured by Parsivel2 [26]. The PSD of falling particles is calculated as follows: N(Di) = nd ∑ j=1 Cij Ai · ∆t · Vj · ∆Di ; N(Di) [mm−1m−3], Di[mm] (1) (1) where N(Di) is the number of particles from Di to Di + ∆Di in a unit volume, and Di, ∆Di are the average and bin spacing of the i-th size bin, respectively. Cij is the number of particles measured in the i-th size bin and the j-th speed bin, and nd is the number of size bins (32 in this study). Vj is the measured time interval (60 s for this instrument), and Ai is the fall rate measured for the j-th speed bin. The effective measurement area of the i-th size bin is calculated as follows: Ai = [180 × (30 −0.5Di)] × 10−6 [m2] (2) (2) The mean fall velocity of particles with a mean diameter Di is calculated as follows: V(Di) = nv ∑ j=1 Vj · Cij/ nv ∑ j=1 Cij [m s−1] (3) (3) where nv is the number of velocity bins (32 for this instrument). The ash fall rate RA was calculated as follows: RA = πρp 6 nd ∑ i=1 (V(Di) · N(Di)·D3 i ); RA [kg m−2s−1] (4) (4) where ρp is the solid density of ash particles. We assumed that ρp = 2.5 g cm−3 (= 2.5 × 103 kg m−3). From Equation (1), RA = π 6 10−9ρp nd ∑ i=1 nv ∑ j=1 D3 i Cij Ai ∆t; RA [kg m−2s−1] (5) (5) If RA is expressed in [mm h−1], If RA is expressed in [mm h−1], RA = 6π 10−4 ρp ρb nd ∑ i=1 nv ∑ j=1 D3 i Cij Ai ∆t; RA [mm h−1] (6) (6) where ρb is the bulk density of ash deposits. We assumed that ρb = 1.25 g cm−3 (= 1.25 × 103 kg m−3). 3.1. Radar Meteorological Approach 3.1. Radar Meteorological Approach In radar meteorology, precipitation PSD is indispensable for quantitative estimation of precipitation and hydrometeor classiﬁcation. Therefore, functions to approximate observed precipitation particles have been proposed in several previous studies. Functional forms of PSD can be used to investigate spatiotemporal variation in the microphysical processes of precipitation clouds, derive theoretical formulae for quantitative precipitation estimates, and simulate precipitation particle scattering. In this study, we applied radar meteorology concepts and techniques to volcanic ash fall phenomena. In this section, we describe volcanic ash PSD using either gamma or normalized gamma functions. In addition, we show that the integrated PSD parameters such as RA, CA, Z, etc. can be expressed in terms of either gamma or normalized gamma PSD models. 2.3. Data Processing The volcanic ash mass concentration CA is deﬁned as the amount of ash particles in a unit volume of air, is calculated as follows: CA = π 6 10−6ρp nd ∑ i=1 nv ∑ j=1 D3 i Cij Ai ∆t Vj ; CA [kg m−3] (7) (7) and the reﬂectivity factor Z is calculated as follows: and the reﬂectivity factor Z is calculated as follows: and the reﬂectivity factor Z is calculated as follows: Z = nd ∑ i=1 nv ∑ j=1 D6 i Cij Ai ∆t Vj ; Z [mm6m−3] (8) (8) Atmosphere 2021, 12, 601 6 of 24 The mass-weighted diameter Dm is a parameter characterizing the normalized PSD, and is calculated as the ratio of the fourth moment of PSD to the third moment, as follows: Dm = nd ∑ i=1 N(Di)D4 i ∆Di nd ∑ i=1 N(Di)D3 i ∆Di ; Dm [mm] (9) (9) Dm is closely related to the median volume particle diameter D0 (Sections 3.2 and 3.4). The normalized PSD intercept parameter Nw is calculated as follows: Dm is closely related to the median volume particle diameter D0 (Sections 3.2 and 3.4). The normalized PSD intercept parameter Nw is calculated as follows: Nw = 44 πρp CA D4m ; Nw [mm−1m−3] (10) (10) The basic parameters of volcanic ash particles described above were directly calculated from PSD data measured by Parsivel2. The basic parameters of volcanic ash particles described above were directly calculated from PSD data measured by Parsivel2. 3.2. Gamma PSD Model Raindrop size distribution is commonly described using the gamma PSD model. In this study, we replaced raindrops with falling ash particles in the gamma PSD model, which is expressed as follows [27,28]: N(D) = N0Dµ exp(−ΛD) (11) (11) where N(D) is the number of ash particles per unit diameter increment within a unit volume, N0 (mm−1−µ m−3) is the intercept parameter characterizing PSD, Λ (mm−1) is the slope parameter, and µ is the shape parameter. D is expressed in mm, and µ, which is normally dimensionless, takes the units of N0. Thus, N0 and µ are not completely independent, such that N0 is affected by the variation of µ and N0 has no physical meaning [29]. Equation (11) can be expressed using the median volume diameter D0, which has a physical meaning and is used in radar meteorology instead of Λ. Thus, N(D) = N0Dµ exp[−G(D/D0)] (12) (12) When Dmax/D0 ≥2.5, and µ ≥−3, the following approximation holds, with an accuracy of ≤0.5% [27]: G = ΛD0 = 3.67 + µ. The median volume diameter D0 is deﬁned as the diameter that divides the total volume of all falling ash particles within a unit volume into two equal parts in the falling ash PSD. Since the computation of D0 is cumbersome, the mass-weighted mean Dm is often used instead of D0. The relationship between Dm and D0 is as follows [27]: When Dmax/D0 ≥2.5, and µ ≥−3, the following approximation holds, with an accuracy of ≤0.5% [27]: G = ΛD0 = 3.67 + µ. The median volume diameter D0 is deﬁned as the diameter that divides the total volume of all falling ash particles within a unit volume into two equal parts in the falling ash PSD. Since the computation of D0 is cumbersome, the mass-weighted mean Dm is often used instead of D0. The relationship between Dm and D0 is as follows [27]: Dm = (4 + µ)/(3.82 + µ)D0 (13) (13) Atmosphere 2021, 12, 601 7 of 24 The value 3.82 in the above equation is attributed to a power law approximation of the fall velocity of ash particles (D ≤2 mm; Table 3). The value 3.67 is typically used for raindrops. From Equation (13), when µ ≥0, the difference between Dm and D0 is less than 9%, and as µ becomes larger, this difference becomes smaller; when µ = 10, the difference is about 2%. 3.2. Gamma PSD Model Using Dm to represent the gamma PSD, the model becomes: N(D) = N0Dµ exp[−(4 + µ)(D/Dm)] (14) N(D) = N0Dµ exp[−(4 + µ)(D/Dm)] (14) The intercept parameter Λ can be expressed as follows: The intercept parameter Λ can be expressed as follows: Λ = (4 + µ)/Dm (15) (15) As a special case of the gamma distribution, µ = 0, As a special case of the gamma distribution, µ = 0, N(D) = N0 exp(−ΛD) = N0 exp[−G(D/D0)] (16) (16) or or N(D) = N0 exp[−4(D/Dm)] (17) (17) This PSD model is a well-known exponential PSD function [30]. Table 3. List of coefﬁcients and exponents of ash particle fall velocity formulae proposed in previous st where Vt is expressed in m s−1 and D is expressed in mm. ﬁcients and exponents of ash particle fall velocity formulae proposed in previous studies. Vt (D) = a Db, ed in m s−1 and D is expressed in mm. exponents of ash particle fall velocity formulae proposed in previous studies. Vt (D) = a Db, Table 3. List of coefﬁcients and exponents of ash particle fall velocity formulae proposed in previous studies. Vt (D) = a Db, where Vt is expressed in m s−1 and D is expressed in mm. Table 3. List of coefﬁcients and exponents of ash particle fall velocity formulae proposed in previous studies. Vt (D) = a Db, where Vt is expressed in m s−1 and D is expressed in mm. a b Note Reference 5.90 0.530 0.25 ≤D < 4 mm (free fall experiments, Parsivel2) Miwa et al. [31] 5.96 0.530 0.25 ≤D < 4 mm (free fall experiments, 2DVD) Suh et al. [10] 4.04 0.830 0.25 ≤D < 2 mm (Sakurajima, Parsivel2, 26,075 particles) Kozono et al. [32] 3.96 0.748 0.25 ≤D < 4 mm (Sakurajima, Parsivel2, 79,170 particles) Kozono [33] 3.18 0.728 0.25 ≤D < 4 mm (Sakurajima, Parsivel2, 63,237 particles) Present study 3.14 0.817 0.25 ≤D < 2 mm (Sakurajima, Parsivel2, 63,237 particles) Present study 3.42 0.371 0.2 ≤D ≤6.0 mm (Mt. St. Helens, 19 March 1982) Harris and Rose [34] 5.02 0.335 0.22 ≤D ≤1.3 mm (Mt. St. Helens, 19 March 1982) Harris and Rose [34] 6.87 1.0 0.02 ≤D ≤0.38 mm (Mt. St. Helens, 18 May 1980) Harris and Rose [34] 4.33 0.437 0.02 ≤D ≤6.0 mm (Mt. St. Helens, all data) Present study (average) 3.3. Normalized Gamma PSD Model 3.3. Normalized Gamma PSD Model 3.3. Normalized Gamma PSD Model Generally, falling ash PSDs ﬂuctuate depending on CA or RA. To investigate the shape of the ash PSD, which is not affected by CA, normalized (scaled) PSD concepts have been proposed [35–40]. The normalized PSD is generally expressed as follows [39]: N(D) = NwF(D/Dm) = NwF(X) (18) (18) where Nw is the normalized intercept parameter, which normalizes N (D), and Dm is the mass-weighted mean diameter, which normalizes D. F is a factor that determines the geometry of the PSD, and X = D/Dm. The normalized PSD expressed in Equation (18) is not a functional form of PSD. In this section, we derive a normalized PSD assuming a gamma function for measured PSD. When N(D) is represented by the gamma function, i.e., Equation (12), the nth moment Mn of PSD is expressed as follows: Mn = Z ∞ 0 DnN(D)dD = N0Γ(n + 1 + µ) Λn+µ+1 (19) (19) Atmosphere 2021, 12, 601 8 of 24 Dm and Nw are expressed as follows, in terms of Mn: Dm and Nw are expressed as follows, in terms of Mn: Dm and Nw are expressed as follows, in terms of Mn: Dm = M4/M3 = (4 + µ)/Λ (20) Dm = M4/M3 = (4 + µ)/Λ (20) Nw = 44 Γ(4) M5 3 M4 4 = N0Dµ m Γ(4 + µ) Γ(4) 44 (4 + µ)4+µ (21) (20) (21) From Equations (14), (18), (20), and (21), From Equations (14), (18), (20), and (21), F(X) = Γ(4) 44 (4 + µ)4+µ Γ(4 + µ) Xµ exp[−(4 + µ)X] (22) (22) Therefore, the normalized gamma PSD can be represented as follows: Therefore, the normalized gamma PSD can be represented as follows: N(D) Nw = Γ(4) 44 (4 + µ)4+µ Γ(4 + µ) D Dm µ exp[−(4 + µ) D Dm ] (23) (23) Importantly, the normalized gamma PSD described in Equation (23) is represented by three independent parameters, Nw, Dm, and µ. Importantly, the normalized gamma PSD described in Equation (23) is represented by three independent parameters, Nw, Dm, and µ. 3.4. Calculation of PSD Parameters 3.4. Calculation of PSD Parameters 3.4. Calculation of PSD Parameters Three parameters of the gamma PSD, N0 (or Nw), Λ (D0 or Dm), and µ can be calculated using the nth moment Mn of the PSD [41,42], as follows: Mn = Z ∞ 0 DnN(D)dD (24) (24) Mn can be calculated by the following equation using PSDs measured by Parsivel2: Mn can be calculated by the following equation using PSDs measured by Parsivel2: Mn = 32 ∑ i=1 Dn i Ni(Di) (25) (25) For convenience of computation, we introduce a variable η, which is deﬁned as the combination of M2, M4, and M6, as follows: For convenience of computation, we introduce a variable η, which is deﬁned as the combination of M2, M4, and M6, as follows: η = M2 4 M2M6 = [Γ(5 + µ)]2 Γ(3 + µ)Γ(7 + µ) = (3 + µ)(4 + µ) (5 + µ)(6 + µ). (26) (26) such that η is a monotonically increasing function with respect to µ. By solving Equation (26), the shape parameter µ of the gamma PSD model can be obtained from the following equation: µ = (7 −11η) −(η2 + 14η + 1)1/2 2(η −1) . (27) (27) If µ is known, the slope parameter Λ of the gamma proﬁle is given by [42]: Λ = M2Γ(5 + µ) M4Γ(3 + µ) 1/2 = M2(4 + µ)(3 + µ) M4 1/2 (28) (28) where Γ(x) = (x −1)Γ(x −1). N0 is calculated as follows: where Γ(x) = (x −1)Γ(x −1). N0 is calculated as follows: N0 = MnΛµ+n+1 Γ(µ + n + 1) (n = 2, 4, or 6) (29) (29) Atmosphere 2021, 12, 601 9 of 24 where N0 can be obtained by specifying any value of n if µ and Λ are obtained. Thus, n can be 2, 4, or 6, and the difference in the calculated value of N0 for each case is about 1%. D0 can be computed as follows if µ and Λ are obtained: where N0 can be obtained by specifying any value of n if µ and Λ are obtained. Thus, n can be 2, 4, or 6, and the difference in the calculated value of N0 for each case is about 1%. 3.4. Calculation of PSD Parameters D0 can be computed as follows if µ and Λ are obtained: D0 = 3.82 + µ Λ (30) (30) The present paper utilizes the complete moment method to calculate the gamma PSD parameters. However, it may be necessary to mention that the calculation of gamma PSD parameters is more complex than the complete moment method when the lower and upper bounds of the integral are ﬁnite in Equation (24), i.e., an incomplete gamma distribution. The effects of Dmin and Dmax on the calculated gamma PSD parameters have been examined assuming an incomplete gamma distribution [42]; the effect of Dmin on the gamma PSD parameters is small, such that Dmin = 0 may be assumed [43,44]. The present paper use the observed Dmax to estimate the nth moment of the PSD. 3.5. Integrated PSD Parameters The gamma PSD parameters obtained by the full moment method in Section 3.4 may then be substituted into these equations. 3.5. Integrated PSD Parameters The integrated PSD parameters include the total number of falling ash particles NT, CA, RA, and Z. The integrated parameters are expressed by PSD moments, as follows: NT = Z Dmax 0 N(D)dD = m0 (31) CA = ρwπ 6 Z Dmax 0 D3N(D)dD = ρwπ 6 m3 (32) RA = ρwπ 6 R Dmax 0 D3vt(D)N(D)dD aρwπ R Dmax D3+bN(D)dD ρwπ m (33) NT = Z Dmax 0 N(D)dD = m0 (31) (31) CA = ρwπ 6 Z Dmax 0 D3N(D)dD = ρwπ 6 m3 (32) (32) RA = ρwπ 6 R Dmax 0 D3vt(D)N(D)dD = aρwπ 6 R Dmax 0 D3+bN(D)dD = ρwπ 6 m3+b (33) (33) where v(D) is the falling velocity of volcanic ash particles of diameter D, calculated as follows: where v(D) is the falling velocity of volcanic ash particles of diameter D, calculated as follows: vt(D) = aDb (34) (34) Z = Z Dmax 0 D6N(D)dD = m6 (35) (35) The integrated PSD parameters deﬁned in Equations (31)–(33) are determined by PSD moments calculated from the measured PSD data, and the relationships among integrated PSD parameters can be obtained by regression analyses of the integrated PSD parameters. If the PSD data are not available, then the relationships among integrated PSD parameters are derived by assuming the functional form of the PSD. Assuming the gamma PSD model, the following relationships are derived: NT = m0 = N0 Λµ+1 γ µ + 1, α Dmax Dm (36) (36) CA = ρwπ 6 m3 = ρwπ 6 N0 Λµ+4 γ µ + 4, α Dmax Dm (37) RA = ρwπ 6 m3+b = ρwπ 6 N0 Λµ+b+4 γ µ + b + 4, α Dmax Dm (38) Z = m6 = N0 Λµ+7 γ µ + 7, α Dmax Dm (39) (37) (38) Z = m6 = N0 Λµ+7 γ µ + 7, α Dmax Dm (39) (39) The gamma PSD parameters obtained by the full moment method in Section 3.4 may then be substituted into these equations. The gamma PSD parameters obtained by the full moment method in Section 3.4 may then be substituted into these equations. Atmosphere 2021, 12, 601 10 of 24 10 of 24 4. Results 4.1. Fall Speed of Volcanic Ash Particles 4.1. Fall Speed of Volcanic Ash Particles The terminal velocity of falling particles is generally determined according to the balance among gravitational, buoyancy, and aerodynamic drag forces [45]. In the present study, we express the particle fall velocity as a power law function of particle diameter, as shown in Equation (33), which is obtained by applying regression analysis to fall velocity data measured using Parsivel2, as shown in Equation (3). g q The distribution of the number of ash particles in the parameter space of fall velocity and diameter for the six selected volcanic eruptions is shown in Figure 3. The total number of samples was 63,237, and most of the observed data were for ash particles, i.e., D ≤2 mm. Particles with diameters ranging from 0.4 to 0.8 mm were the most frequent. The fall velocities corresponding to these particles were widely distributed, from 0.2 to 4 ms−1, with larger particles occurring less frequently but with greater variance. This variation may be caused by Parsivel2 measurement error or particle density variation. The falling velocity of particles increased in proportion to the square of the particle density [45]. Another reason for the large scatter observed in Figure 3 may be the particle shape, which determines the drag force acting on the particle [10]. The ﬂatter the particle shape, the slower the falling velocity. Figure 3. Density plots of the particle size and fall velocity of volcanic ash particles for six selected eruptions of the Sakurajima volcano. Data were collected by Parsivel2. Color scale indicates drop counts on a log scale. Line indicates the regression curve based on a power law. Figure 3. Density plots of the particle size and fall velocity of volcanic ash particles for six selected eruptions of the Sakurajima volcano. Data were collected by Parsivel2. Color scale indicates drop counts on a log scale. Line indicates the regression curve based on a power law. Figure 3. Density plots of the particle size and fall velocity of volcanic ash particles for six selected eruptions of the Sakurajima volcano. Data were collected by Parsivel2. Color scale indicates drop counts on a log scale. Line indicates the regression curve based on a power law. The largest particle observed in the present study was 4 mm in diameter. 4.1. Fall Speed of Volcanic Ash Particles The power law equation for the fall velocity of all data shown in Figure 3 is: vt = 3.18D0.728; vt [ms−1], D [mm] (40) (40) For volcanic ash particles (D ≤2 mm), the fall velocity equation is: For volcanic ash particles (D ≤2 mm), the fall velocity equation is: vt = 3.14D0.817; vt [ms−1], D [mm] (41) (41) which is quite similar to Equation (40). which is quite similar to Equation (40). Table 3 summarizes the fall velocity equation derived in the present study and those proposed in previous studies. The equations for fall velocity versus diameter listed in Table 3 are shown in Figure 4. Among all curves shown in Figure 4, the formulae derived Atmosphere 2021, 12, 601 11 of 24 from experiments of ash particle free fall from a height of 17 m [10,31] give the largest fall velocity for the same diameter, likely because the fall velocity was accelerated by the ﬁngering phenomenon [46,47]. Parsivel2, installed on Sakurajima, was used to observe the falling velocity of actual ash particles. [11,32]; the falling velocity of ash particles for a total of 76 eruptions from 2014 to 2016 was described as a power law (Figure 4) derived from observed data [33] and previously developed theoretical equations [11,34,48]. The power law formulae obtained in the present study provided the lowest fall velocity for the same diameter, and smaller than with the theoretical equations [34] in which the particle density was assumed to be 2500 (kg m−3). Figure 4. Relationships between ash particle fall velocity and diameter proposed in previous studies and the present one. Figure 4. Relationships between ash particle fall velocity and diameter proposed in previous studies and the present one. 4.2.1. Temporal Changes in PSD 4.2.1. Temporal Changes in PSD Changes in the PSDs of the six selected eruptions over time, as eruption clouds passed over the Parsivel2 device, are shown in Figure 5. The maximum RA (11.7 mm h−1) was observed during eruption 4. The number of particles observed per unit volume (NT) ranged from 10 to 103 m−3. RA and log10NT were somewhat correlated. Particles with a maximum diameter of 3.75 mm were observed in eruptions 2b and 3. Dmax was not necessarily correlated with RA. In all eruptions except eruption 1, large values of Dmax were observed immediately after the onset of ash fall (Figure 5). Dmax and Dm were positively correlated, and Dmax was negatively correlated with both log10NT and log10 Nw. These new ﬁndings on the PSD of ash particles will be examined in detail in the following sections. A temporal change in the PSD at a certain point is considered to reﬂect a spatial change in PSD within the eruption cloud. Therefore, it is necessary to know which part of the eruption cloud was observed by Parsivel2 when discussing a temporal change in the PSD. The distribution of the time-integrated reﬂectivity factor in eruption clouds during eruption 6 is shown in Figure 6. We also obtained the trajectories of the centroids of the eruption clouds (data not shown). Calculations of time-integrated radar reﬂectivity and the trajectories are based on PPI data at an elevation angle of 1.7◦. Parsivel2 data for ash particles in the central part of eruption cloud as it passed over the observation site are shown in Figure 5. Unfortunately, there is no Parsivel2 site in the area where the accumulated reﬂectivity factor reached a maximum, since this area is 2 km from the vent and is therefore restricted. 12 of 24 12 of 24 Atmosphere 2021, 12, 601 Figure 5. Temporal changes in gamma PSD parameters observed by Parsivel2 during eruption 6 (16 July 2018). (a) Ash fall rate RA. (b) Total number of volcanic ash particles NT. (c) Maximum diameter Dmax. (d) Normalized intercept parameter Nw. (e) Mass-weighted diameter Dm. Figure 5. Temporal changes in gamma PSD parameters observed by Parsivel2 during eruption 6 (16 July 2018). (a) Ash fall rate RA. (b) Total number of volcanic ash particles NT. (c) Maximum diameter Dmax. (d) Normalized intercept parameter Nw. (e) Mass-weighted diameter Dm. Figure 6. 4.2.1. Temporal Changes in PSD Plan position indicator (PPI) image of time-integrated radar reﬂectivity for eruption clouds formed in eruption 6 (16 July 2018). PPI images were obtained every 2 min at an elevation angle of 1.7◦. Circle indicates the location of Parsivel2; triangle indicates the summit of Minami-dake. Figure 6. Plan position indicator (PPI) image of time-integrated radar reﬂectivity for eruption clouds formed in eruption 6 (16 July 2018). PPI images were obtained every 2 min at an elevation angle of 1.7◦. Circle indicates the location of Parsivel2; triangle indicates the summit of Minami-dake. 4.2.2. Gamma PSD Parameters Temporal changes in the observed PSD proﬁles for eruption 6 are shown in Figure 7, as a representative example. Two ﬁtting curves of the gamma PSD function are superimposed on each set of observed PSD data; one curve was obtained by the momentum method, and the other curve, by non-linear regression analysis. It should be noted that non-linear regression analysis must be carried out to the logarithmic form of the gamma PSD formula because N(D) distributes over a wide range (from 100 to 104). A good ﬁt was obtained for all samples, except at 15:54 LST, when large particles were observed. The gamma PSD obtained by the momentum method was approximately exponential, while the gamma PSD obtained by non-linear regression analysis exhibited an upward convex shape that ﬁtted small particles. It should be noted that the results obtained by the two methods employed agree quite well for PSDs at 16:00 LST to 16:04 LST, a period during which only particles smaller than 2 mm were observed. The results suggest that the difference in the gamma PSD Atmosphere 2021, 12, 601 13 of 24 functional forms obtained by non-linear regression analysis and the momentum method is acceptable. We analyzed all the PSD data obtained in order to quantitatively conﬁrm the results, which are shown in the discussion section of the present paper. Figure 7. Temporal changes in measured PSD for eruption 6 (16 July 2018). Solid lines and dashed lines are the gamma PDS ﬁtting curves for the measured PSD data by the moment method and non-linear regression analysis, respectively. Figure 7. Temporal changes in measured PSD for eruption 6 (16 July 2018). Solid lines and dashed lines are the gamma PDS ﬁtting curves for the measured PSD data by the moment method and non-linear regression analysis, respectively. To clarify the statistical characteristics of the PSDs of volcanic ash particles, we in- vestigated the frequency distribution and probability density function of four parameters (log10N0, Λ, D0, and µ) of gamma PSDs, and of the additional PSD parameters Dmax and NT observed in all eruptions. The results are shown in Figure 8. The six parameters (log10N0, D0, Λ, µ, Dmax, and NT) showed modes (standard deviation [SD]) of 8.8 (9.6), 0.65 (0.29), 13.4 (25.0), 8.3 (13.7), 0.97 (0.679), and 182 (380), respectively. It interesting that each of three parameters (log10N0, Λ, µ) had a wider range compared with that of precipitation particles. 4.2.2. Gamma PSD Parameters Other descriptive statistics, such as the maximum, median, and skewness, are summarized in Table 4. al values of gamma, normalized, and integrated particle size distribution (PSD) parameters. ρb = 1.25 × 103 = 2.50 × 103 kg m−3 were assumed for the estimation of RA. Table 4. Statistical values of gamma, normalized, and integrated particle size distribution (PSD) parameters kg m−3 and ρp = 2.50 × 103 kg m−3 were assumed for the estimation of RA. g ρp g Parameter Unit Mode Med Mean SD Max Skew D10 D90 log10 N0 mm−1 m−3 8.83 9.64 15.8 9.64 47.9 0.581 4.79 28.2 L mm−1 13.4 27.2 33.5 25.0 118 0.742 6.05 65.4 D0 mm 0.65 0.69 0.79 0.292 2.13 1.88 0.54 1.21 m – 8.3 17.1 18.9 13.7 67.3 0.661 3.10 36.8 Dmax mm 0.97 1.19 1.42 0.679 3.75 1.42 0.81 2.36 log10Nw mm−1 m−3 4.60 4.42 4.15 0.704 5.15 −0.875 3.07 4.87 Dm mm 0.853 1.19 0.853 0.350 2.36 1.83 0.559 139 NT m−3 182 252 381 380 1788 1.75 26 998 RA mm h−1 0.52 1.08 2.01 2.17 11.7 1.64 0.23 5.43 CA g m−3 0.0931 0.180 0.225 0.189 0.845 1.21 0.0323 0.478 10log10Z dBZ 17.1 18.2 19.3 7.43 36.9 0.087 9.28 28.8 Atmosphere 2021, 12, 601 14 of 24 Figure 8. Histogram and probability density function of gamma PSD parameters including the (a) log of the intercept parameter (log10N0), (b) median volume diameter (D0), (c) slope parameter (Λ), (d) shape parameter (µ), (e) maximum particle diameter (Dmax), and (f) total particle number (NT). Figure 8. Histogram and probability density function of gamma PSD parameters including the (a) log of the intercept parameter (log10N0), (b) median volume diameter (D0), (c) slope parameter (Λ), (d) shape parameter (µ), (e) maximum particle diameter (Dmax), and (f) total particle number (NT). The main objective of radar remote sensing is determining the physical quantities of eruption clouds from measured radar parameters. Normally, the number of physical quantities, which are unknowns, is larger than the number of parameters that can be measured by radar. Therefore, the number of unknowns is reduced by theoretically or empirically deﬁning relationships between unknown parameters. If two parameters are strongly correlated, then the obtained relationship will be incorporated into a weather radar monitoring system for eruption clouds. Theoretical interpretations of such correlations are useful for understanding the microphysical processes of eruption clouds. 4.2.2. Gamma PSD Parameters Clear correlations were detected between gamma PSD parameters in this study (Figure 9), which suggests that the gamma PSD model well describes the observed volcanic ash PSDs. The relationships between gamma PSD parameters are summarized in Table 5. Thus, if one gamma PSD parameter is obtained, the remaining parameters can be estimated using the information listed in Table 5. A scatter plot of Dmax and µ is also shown in Figure 9; the shape parameter µ approaches 0 (i.e., exponential PSD) when Dmax > ~3, and becomes large (i.e., upward concave PSD) when Dmax is small. Thus, once Dmax is given for an initial condition of the PSD, µ is determined by the µ–Dmax relationship. Next, N0 and Λ are estimated by the N0–µ and Λ–µ relationships, respectively. Table 5. Summary of relationships among gamma PSD parameters. The numbers in the brackets are the correlation coefﬁcient and the root-mean-square error, respectively. log10N0 Λ µ Dmax log10N0 - log10N0 = 0.383 Λ + 2.942 (0.995, 0.91) log10N0 = 0.684 µ + 2.89 (0.973, 2.211) log10N0 = 19.67 Dmax −1.606 (0.920, 3.784) Λ Λ = 2.585 log10N0 −7.308 (0.995, 2.361) - Λ = 1.751 µ + 0.469 (0.960, 7.005) Λ = 42.23 Dmax−2.046 (0.937, 8.76) µ µ = 1.386 log10N0 −3.02 (0.974, 3.148) µ = 0.527 Λ + 1.222 (0.960, 3.842) - µ = 23.87 Dmax−1.713 (0.853, 7.178) Dmax Dmax = 5.863 log10N0 − 0.595 (0.922, 0.263) Dmax = 4.862 Λ−0.418 (0.937, 0.238) Dmax = 3.018 µ−0.314 (0.779, 0.393) - Table 5. Summary of relationships among gamma PSD parameters. The numbers in the brackets are the correlation coefﬁcient and the root-mean-square error, respectively. 15 of 24 Atmosphere 2021, 12, 601 Figure 9. Scatter plots of gamma PSD parameters: (a) Λ vs. log10N0, (b) Λ vs. µ, (c) Λ vs. Dmax, (d) µ vs. log10N0, (e) µ vs. Dmax, and (f) Dmax vs. log10N0. Figure 9. Scatter plots of gamma PSD parameters: (a) Λ vs. log10N0, (b) Λ vs. µ, (c) Λ vs. Dmax, (d) µ vs. log10N0, (e) µ vs. Dmax, and (f) Dmax vs. log10N0. 4.2.3. Normalized PSD Figure 10 is a scatter plot showing PSDs before and after normalization. The normal- ized PSD parameters (Nw and Dm) were calculated directly from the PSD data measured by Parsivel2, without assuming a functional form of the PSD. Figure 10a shows the PSD before normalization, which is characterized by wide scattering due to variation in ash fall strength, such that the characteristic distribution could not be observed. After normal- ization, the PSDs converged to reduce the scatter (Figure 10b), allowing us to observe the form of the distribution. The shape parameter µ was determined by least-squares ﬁtting of the measured N(D)/Nw data according to Equation (23). It may be interesting to examine the associations between Nw or Dm with the type and magnitude of volcanic eruption in a future study, using larger sample sizes. Figure 10. Comparison of PSD before and after normalization: (a) N(D) vs. D for the six selected volcanic eruptions measured by Parsivel2 and (b) corresponding plot of N(D)/Nw vs. D/Dm. Figure 10. Comparison of PSD before and after normalization: (a) N(D) vs. D for the six selected volcanic eruptions measured by Parsivel2 and (b) corresponding plot of N(D)/Nw vs. D/Dm. Atmosphere 2021, 12, 601 16 of 24 Despite the small sample sizes in the present study, they were useful for determining the statistical characteristics of normalized gamma PSD parameters. Figure 11 shows the frequency distributions and probability density functions of the two parameters (Dm and log10Nw) used to normalize the PSD. The shape parameter µ is shown in Figure 8d. The modes (SD) of Dm and log10Nw of the normalized gamma PSD parameters were 0.656 (0.350) and 4.60 (0.704), respectively, and the mode (SD) of µ was 8.31 (13.7). Figure 11. The same as Figure 8, but for normalized PSD parameters including (a) log10Nw and (b) Dm. Figure 11. The same as Figure 8, but for normalized PSD parameters including (a) log10Nw and (b) Dm. 4.2.4. Quantitative Ash Fall Estimation Using Normalized PSD Parameters Next, we examined correlations between RA and gamma PSD parameters (Nw, Dm, and µ). Figure 12 shows a scatter plot of RA/Nw and Dm. Applying nonlinear regression analysis, we obtained the following power law equation: RA/Nw = 2.54 × 10−4Dm4.97 (42) RA/Nw = 2.54 × 10−4Dm4.97 (42) Figure 12. The same as Figure 9, but for normalized PSD parameters including (a) Dm vs. log10Nw, (b) Dm vs. µ, (c) Dm vs. 4.2.3. Normalized PSD Dmax, (d) log10Nw vs. µ, and (e) log10Nw vs. Dmax. By rearranging Equation (42), we obtained: (42) Figure 12. The same as Figure 9, but for normalized PSD parameters including (a) Dm vs. log10Nw, (b) Dm vs. µ, (c) Dm vs. Dmax, (d) log10Nw vs. µ, and (e) log10Nw vs. Dmax. By rearranging Equation (42), we obtained: RA = 2.54 × 10−4 Dm4.97Nw (43) (43) Atmosphere 2021, 12, 601 17 of 24 17 of 24 The power law equations for the upper and lower 95% confidence limits of Equation (43) are as follows: RA = 2.72 × 10−4 Dm5.07Nw; 95% upper limit (44) RA = 2.36 × 10−4 Dm4.87Nw; 95% lower limit (45) (44) (45) (45) In Figure 12, the correlation coefﬁcient (root-mean-square error) between RA/Nw and Dm was 0.991 (0.000177), which was higher (lower) than that between RA and Z, as described in the next section. This result suggests that Equation (42) provides an advanced method for estimating RA if Nw and Dm are estimated with a sufﬁcient degree of accuracy. For precipitation phenomena, two-frequency radar observations based on satellite remote sensing [49] and dual polarization radar observation [50] have been proposed to estimate Nw and Dm. Similar techniques may be available for ash fall phenomena, and should be explored in a future study. As physical quantities that represent the microphysical processes within precipitation clouds, logNw and Dm have also been used to distinguish precipitation types (stratiform or convective) [51,52]. Similarly, Dm and Nw are expected to provide new insight into volcanic eruption clouds and their microphysical processes in future studies [53–55]. 4.3. Conventional Relationship for Quantitative Ash Fall Estimation 4.3. Conventional Relationship for Quantitative Ash Fall Estimation 4.3. Conventional Relationship for Quantitative Ash Fall Estimation Note that RA (mm h−1), CA (g m−3), Z (mm6 m−3). R: correlation coefﬁcient, RMSE: root mean square error. ρb = 1.25 × 103 kg m−3 and ρp = 2.50 × 103 kg m−3 were assumed for the estimation of RA. Case Site Sampling Period (LST) (min) RA–Z Relationship (R, RMSE) CA–Z Relationship (R, RMSE) Range 1 NABE 08:22–08:35, 15 May 2018 14 RA = 8.62 × 10−2 Z 0.809 (0.963, 0.300) CA = 1.51 × 10−2 Z 0.680 (0.912, 0.046) 0 ≤Z ≤1 × 102 2 HART 20:03–20:11, 22 May 2018 9 RA = 5.44 × 10−2 Z 0.595 (0.910, 0.907) CA = 0.629 × 10−2 Z 0.569 (0.950, 0.058) 0 ≤Z ≤2 × 103 HIKP 20:00–20:14, 22 May 2018 12 RA = 3.03 × 10−2 Z 0.826 (0.977, 0.397) CA = 0.496 × 10−2 Z 0.770 (0.980, 0.043) 0 ≤Z ≤1 × 103 3 KURP 08:09–08:13, 30 May 2018 4 RA = 14.6 × 10−2 Z 0.522 (0.959, 0.613) CA = 1.72 × 10−2 Z 0.423 (0.903, 0.055) 0 ≤Z ≤1 × 103 4 HART 12:03–12:20, 10 June 2018 12 RA = 9.45 × 10−2 Z 0.568 (0.972, 0.929) CA = 4.49× 10−2 Z 0.294 (0.899, 0.073) 0 ≤Z ≤5 × 103 5 SBTT 07:35–08:21, 16 June 2018 31 RA = 3.55 × 10−2 Z 0.778 (0.917, 0.339) CA = 1.20 × 10−2 Z 0.692 (0.960, 0.036) 0 ≤Z ≤4 × 102 6 AKAM 15:53–16:12, 16 July 2018 18 RA = 10.0 × 10−2 Z 0.653 (0.851, 1.123) CA = 0.553 × 10−2 Z 0.782 (0.913, 0.100) 0 ≤Z ≤7 × 102 All - - 88 RA = 22.8 × 10−2 Z 0.436 (0.819, 1.247) CA = 6.56 × 10−2 Z 0.261 (0.634, 0.147) 0 ≤Z ≤5 × 103 Figure 14. Quantitative ash fall estimation based on the relationships among integrated PSD param- eters including (a) RA–Z, (b) RA/Nw–Dm, where RA, Nw, and Dm were calculated using PSD data obtained by Parsivel2. Figure 14. Quantitative ash fall estimation based on the relationships among integrated PSD param- eters including (a) RA–Z, (b) RA/Nw–Dm, where RA, Nw, and Dm were calculated using PSD data obtained by Parsivel2. Figure 14. Quantitative ash fall estimation based on the relationships among integrated PSD param- eters including (a) RA–Z, (b) RA/Nw–Dm, where RA, Nw, and Dm were calculated using PSD data obtained by Parsivel2. 4.3. Conventional Relationship for Quantitative Ash Fall Estimation Figure 13 shows the frequency and probability density distributions of the three integrated PSD parameters (RA, CA, and 10log10Z). In this study, the modal values (SD) of RA, CA, and 10log10Z of the integrated PSD parameters were 0.52 (2.17), 0.0931 (0.189), and 17.1 (7.48), respectively. Figure 13. The same as Figure 8, but for integrated PSD parameters including (a) RA and (b) CA. Figure 13. The same as Figure 8, but for integrated PSD parameters including (a) RA and (b) CA. Next, we investigated the correlations among these integral parameters. In particular, the relationship between RA and Z is crucial as a practical equation for estimating the RA from weather radar observations. The CA–Z relationship is used to estimate the CA of volcanic ash. These relationships were derived from regression analyses of observed data for each eruption, and are summarized in Table 6. A scatter plot of RA and Z values obtained from Parsivel2 observations for all eruptions is shown in Figure 14. The RA–Z relationship was deﬁned using the nonlinear least-squares method, as follows: RA = 22.8 × 10−2 Z0.436; RA [mm h−1], Z [mm6m−3] (46) (46) The RA–Z relationships for the upper and lower 95% conﬁdence limits are as follows: RA = 31.0 × 10−2 Z0.489; 95% upper limit (47) RA = 14.6 × 10−2 Z0.383; 95% lower limit (48) RA = 31.0 × 10−2 Z0.489; 95% upper limit RA = 31.0 × 10−2 Z0.489; 95% upper limit (47) RA = 14.6 × 10−2 Z0.383; 95% lower limit (48) (47) (48) Atmosphere 2021, 12, 601 18 of 24 It must be mentioned that the R and RMSE for all eruptions are not as good as those of an individual eruption because the PSDs are changeable depending on the eruption case, which is similar to the variation of PSD in precipitation. Table 6. RA–Z and CA–Z relationships obtained from regression analyses of PSD data measured by Parsivel2. Note that RA (mm h−1), CA (g m−3), Z (mm6 m−3). R: correlation coefﬁcient, RMSE: root mean square error. ρb = 1.25 × 103 kg m−3 and ρp = 2.50 × 103 kg m−3 were assumed for the estimation of RA. 4.3. Conventional Relationship for Quantitative Ash Fall Estimation Case Site Sampling Period (LST) (min) RA–Z Relationship (R, RMSE) CA–Z Relationship (R, RMSE) Range 1 NABE 08:22–08:35, 15 May 2018 14 RA = 8.62 × 10−2 Z 0.809 (0.963, 0.300) CA = 1.51 × 10−2 Z 0.680 (0.912, 0.046) 0 ≤Z ≤1 × 102 2 HART 20:03–20:11, 22 May 2018 9 RA = 5.44 × 10−2 Z 0.595 (0.910, 0.907) CA = 0.629 × 10−2 Z 0.569 (0.950, 0.058) 0 ≤Z ≤2 × 103 HIKP 20:00–20:14, 22 May 2018 12 RA = 3.03 × 10−2 Z 0.826 (0.977, 0.397) CA = 0.496 × 10−2 Z 0.770 (0.980, 0.043) 0 ≤Z ≤1 × 103 3 KURP 08:09–08:13, 30 May 2018 4 RA = 14.6 × 10−2 Z 0.522 (0.959, 0.613) CA = 1.72 × 10−2 Z 0.423 (0.903, 0.055) 0 ≤Z ≤1 × 103 4 HART 12:03–12:20, 10 June 2018 12 RA = 9.45 × 10−2 Z 0.568 (0.972, 0.929) CA = 4.49× 10−2 Z 0.294 (0.899, 0.073) 0 ≤Z ≤5 × 103 5 SBTT 07:35–08:21, 16 June 2018 31 RA = 3.55 × 10−2 Z 0.778 (0.917, 0.339) CA = 1.20 × 10−2 Z 0.692 (0.960, 0.036) 0 ≤Z ≤4 × 102 6 AKAM 15:53–16:12, 16 July 2018 18 RA = 10.0 × 10−2 Z 0.653 (0.851, 1.123) CA = 0.553 × 10−2 Z 0.782 (0.913, 0.100) 0 ≤Z ≤7 × 102 All - - 88 RA = 22.8 × 10−2 Z 0.436 (0.819, 1.247) CA = 6.56 × 10−2 Z 0.261 (0.634, 0.147) 0 ≤Z ≤5 × 103 Figure 14. Quantitative ash fall estimation based on the relationships among integrated PSD param- eters including (a) RA–Z, (b) RA/Nw–Dm, where RA, Nw, and Dm were calculated using PSD data obtained by Parsivel2. 5. Discussion In radar meteorology, various mathematical formulas for ‘precipitation particles’ have been proposed by researchers. These include exponential [30], gamma [27,28], log- normal [56,57], Poisson [58], and Weible [59]. The present study assumed a gamma PSD Atmosphere 2021, 12, 601 18 of 24 18 of 24 It must be mentioned that the R and RMSE for all eruptions are not as good as those of an individual eruption because the PSDs are changeable depending on the eruption case, which is similar to the variation of PSD in precipitation. Table 6. RA–Z and CA–Z relationships obtained from regression analyses of PSD data measured by Parsivel2. Note that RA (mm h−1), CA (g m−3), Z (mm6 m−3). 4.3. Conventional Relationship for Quantitative Ash Fall Estimation R: correlation coefﬁcient, RMSE: root mean square error. ρb = 1.25 × 103 kg m−3 and ρp = 2.50 × 103 kg m−3 were assumed for the estimation of RA. Case Site Sampling Period (LST) (min) RA–Z Relationship (R, RMSE) CA–Z Relationship (R, RMSE) Range 1 NABE 08:22–08:35, 15 May 2018 14 RA = 8.62 × 10−2 Z 0.809 (0.963, 0.300) CA = 1.51 × 10−2 Z 0.680 (0.912, 0.046) 0 ≤Z ≤1 × 102 2 HART 20:03–20:11, 22 May 2018 9 RA = 5.44 × 10−2 Z 0.595 (0.910, 0.907) CA = 0.629 × 10−2 Z 0.569 (0.950, 0.058) 0 ≤Z ≤2 × 103 HIKP 20:00–20:14, 22 May 2018 12 RA = 3.03 × 10−2 Z 0.826 (0.977, 0.397) CA = 0.496 × 10−2 Z 0.770 (0.980, 0.043) 0 ≤Z ≤1 × 103 3 KURP 08:09–08:13, 30 May 2018 4 RA = 14.6 × 10−2 Z 0.522 (0.959, 0.613) CA = 1.72 × 10−2 Z 0.423 (0.903, 0.055) 0 ≤Z ≤1 × 103 4 HART 12:03–12:20, 10 June 2018 12 RA = 9.45 × 10−2 Z 0.568 (0.972, 0.929) CA = 4.49× 10−2 Z 0.294 (0.899, 0.073) 0 ≤Z ≤5 × 103 5 SBTT 07:35–08:21, 16 June 2018 31 RA = 3.55 × 10−2 Z 0.778 (0.917, 0.339) CA = 1.20 × 10−2 Z 0.692 (0.960, 0.036) 0 ≤Z ≤4 × 102 6 AKAM 15:53–16:12, 16 July 2018 18 RA = 10.0 × 10−2 Z 0.653 (0.851, 1.123) CA = 0.553 × 10−2 Z 0.782 (0.913, 0.100) 0 ≤Z ≤7 × 102 All - - 88 RA = 22.8 × 10−2 Z 0.436 (0.819, 1.247) CA = 6.56 × 10−2 Z 0.261 (0.634, 0.147) 0 ≤Z ≤5 × 103 It must be mentioned that the R and RMSE for all eruptions are not as good as those of an individual eruption because the PSDs are changeable depending on the eruption case, which is similar to the variation of PSD in precipitation. Table 6. RA–Z and CA–Z relationships obtained from regression analyses of PSD data measured by Parsivel2. Note that RA (mm h−1), CA (g m−3), Z (mm6 m−3). R: correlation coefﬁcient, RMSE: root mean square error. ρb = 1.25 × 103 kg m−3 and ρp = 2.50 × 103 kg m−3 were assumed for the estimation of RA. Table 6. RA–Z and CA–Z relationships obtained from regression analyses of PSD data measured by Parsivel2. 5. Discussion In radar meteorology, various mathematical formulas for ‘precipitation particles’ have been proposed by researchers. These include exponential [30], gamma [27,28], log- normal [56,57], Poisson [58], and Weible [59]. The present study assumed a gamma PSD model for ‘volcanic ash particles’, for the following two reasons. First, gamma PS models have been used widely in previous radar meteorology studies, because the model is relatively simple and its error structure has been investigated [43]. Second, we can compare the results of our analysis with those of previous studies using the same PSD model. The differences in gamma PSD parameters between precipitation particles and volcanic ash Atmosphere 2021, 12, 601 19 of 24 19 of 24 particles, if any, can be used to discriminate eruption clouds from precipitation clouds, one of the goals of monitoring volcanic ash falls using radar. particles, if any, can be used to discriminate eruption clouds from precipitation clouds, one of the goals of monitoring volcanic ash falls using radar. However, when the gamma PSD model is applied to volcanic ash particles, one point remains unclear. ‘Does the gamma PSD model represent observed volcanic ash PSD?’ This question results from the fact that validation studies have been limited due to the lack of observed ash PSD. In this section, we discuss the validity of the gamma PSD model of volcanic ash using PSD data obtained by Parsivel2. First, we examined the validity of the momentum method used to calculate the gamma PSD. As we have already shown in Figure 7 in this paper, the gamma N(D) obtained by the momentum method agreed with that obtained by non-linear regression analysis. However, as the number of analyzed PSD samples was limited in Figure 7, we examined all PSD samples obtained in six eruption cases. It should be mentioned again here that non-linear regression analysis must be executed not to N(D) but to log10N(D). If we used linear N(D) for the analysis, the obtained gamma PSD regression curve would represent only small particles that have large N(D) values. Figure 15 shows comparisons of gamma PSD parameters (N0, µ, Λ) calculated by the momentum method and non-linear regression analysis. Regression lines obtained were as follows. Log10 N0_mom = 0.934 × log10 N0_reg + 1.47, µ_mom = 0.904 × µ_reg + 2.14, Λ_mom = 0.959 × Λ_reg + 2.72. 5. Discussion where, the subscripts ‘mom’ and ‘reg’ in the variables mean the momentum method and regression analysis, respectively. The correlation coefﬁcient R (the root mean square error RMSE) for N0, µ, and Λ were 0.962 (2.23), 0.940 (4.02), and 0.964 (5.38), respectively. We can conﬁrm from these statistical values that the gamma PSD parameters estimated by the moment method agreed with those obtained by non-linear regression analysis. VIEW Figure 15. Comparison of gamma PSD parameters obtained from the complete momentu method and non-linear regression analysis. (a) log N0, (b) μ, and (c) Λ. Figure 15. Comparison of gamma PSD parameters obtained from the complete momentum method and non-linear regression analysis. (a) log N0, (b) µ, and (c) Λ. Figure 15. Comparison of gamma PSD parameters obtained from the complete momentu method and non-linear regression analysis. (a) log N0, (b) μ, and (c) Λ. Figure 15. Comparison of gamma PSD parameters obtained from the complete momentum method and non-linear regression analysis. (a) log N0, (b) µ, and (c) Λ. Atmosphere 2021, 12, 601 20 of 24 mentum Next, we evaluated the validity of the gamma PSD model itself, which is in turn based on the complete momentum method. The result is shown in Figure 16a. Comparing N(D)_g based on the gamma model with N (D)_o observed by a Parsivel2, we obtained N (D)_g = 0.917 × N (D)_o + 0.175. The R and RMSE were 0.825 and 0.554, respectively. According to Figure 16b, most of the residuals of N (D)_g were within ±10% of their values. Given that the value of N (D) ranged from 100 to 104, these statistical values conﬁrmed that the gamma PSD model is appropriate for describing volcanic ash PSD. Next, we evaluated the validity of the gamma PSD model itself, which is based on the complete momentum method. The result is shown in Figure 16a. Com N(D)_g based on the gamma model with N (D)_o observed by a Parsivel2, we obta (D)_g = 0.917 × N (D)_o + 0.175. The R and RMSE were 0.825 and 0.554, respe According to Figure 16b, most of the residuals of N (D)_g were within ±10% o values. Given that the value of N (D) ranged from 100 to 104, these statistical confirmed that the gamma PSD model is appropriate for describing volcanic ash P Figure 16. (a) Comparison of the gamma PSD model and observed PSDs. 5. Discussion The solid line rep the regression line, and the dashed lines are 95% confidence limits. (b) The residual of gamm 0 1 2 3 4 log 10 N(D) o -1 0 1 2 3 4 log 10 N(D) g (a) 0 1 2 3 4 log 10 N(D) g -2 -1 0 1 2 Residual, log 10 N(D) g (b) Figure 16. (a) Comparison of the gamma PSD model and observed PSDs. The solid line represents the regression line, and the dashed lines are 95% conﬁdence limits. (b) The residual of gamma N(D) expressed on a logarithmic scale. The subscripts ‘g’ and ‘o’ in the variables mean the gamma PSD model and observed PSD, respectively. 0 1 2 3 4 log 10 N(D) o -1 0 1 2 3 4 log 10 N(D) g (a) Figure 16. (a) Comparison of the gamma PSD model and observed PSDs. The solid line rep the regression line, and the dashed lines are 95% confidence limits. (b) The residual of gamm Figure 16. (a) Comparison of the gamma PSD model and observed PSDs. The solid line represents the regression line, and the dashed lines are 95% conﬁdence limits. (b) The residual of gamma N(D) expressed on a logarithmic scale. The subscripts ‘g’ and ‘o’ in the variables mean the gamma PSD model and observed PSD, respectively. 6. Summary In the present study, we examined the characteristics of the PSD of volcanic ash particles and the relationships among PSD parameters. We used a total of 166 PSD samples collected by Parsivel2 during six explosive eruptions of the Sakurajima volcano in 2018. The PSD data were analyzed using methods developed for use in radar meteorology. We proposed gamma and normalized PSD models to describe the PSDs of volcanic ash particles. The observed PSD data were well-described by both models. Strong correlations among the gamma PSD parameters (log10N0, Λ, and µ) were found and relationships between the gamma PSD parameters were derived. Interestingly, the µ–Rmax relationship showed that µ changes inversely with Rmax. When Dmax was large, µ approached zero; i.e., the PSD became exponential. These results could be applied to set up initial PSD conditions for a volcanic ash transport and diffusion model. It must be noted that the range of each of the gamma PSD parameters (log10N0, Λ, and µ) of ash particles was wider than that of precipitation particles. The relationships between gamma PSD parameters can be used to solve the inverse problem in radar meteorology, i.e., the retrieval of PSD parameters from weather radar observations. In precipitation studies, D0 (or Λ) can be estimated from the differential reﬂectivity ZDR, which is measured by polarimetric radar, and N0 is estimated from D0 and observed Z, assuming that µ = 0 [60]. This method is based on the relationship between Atmosphere 2021, 12, 601 21 of 24 21 of 24 raindrop shape and diameter, such that ﬂatter raindrops are associated with larger raindrop diameters. Recently, volcanic ash particle shape has been studied using 2DVD, which was designed to measure the shape of a falling raindrop. The classiﬁcation of ash particle shapes can then be used to identify the volcanic eruption type [61]. To date, a clear relationship between particle shape and diameter has not been found for volcanic ash particles. Further studies of volcanic ash particle shape are required to establish a method for weather radar retrieval of ash particle PSD. In this study, we also proposed a normalized PSD model, which does not require consideration of the PSD variation associated with ash fall intensity. We found a clear correlation between RA/Nw and Dm, and derived its power law functional relationship. 6. Summary Once the two parameters Nw and Dm are retrieved from radar measurements, RA can be estimated from the RA/Nw–Dm relationship proposed in this study. However, further theoretical and observational studies are needed to retrieve parameters Nw and Dm; for example, scattering simulations based on observed PSD could be used to establish theo- retical relationships between Nw and Dm, and polarimetric radar parameters such as ZDR and KDP. Polarimetric radar observations of volcanic ash particles are necessary to validate such theoretical relationships. p Finally, we proposed conventional formulae for estimating the RA and CA. The RA–Z relationship was derived through regression analysis and the CA–Z relationship was calculated from measured PSD data. Although these relationships have large errors associated with instantaneous monitoring of ash fall caused by spatiotemporal variation in PSD, they are convenient and applicable for radar ash fall monitoring. It should be noted that the relationships obtained in this study were derived from a limited number of data. Thus, further analysis using larger PSD datasets is necessary. Author Contributions: Conceptualization, M.M.; methodology, M.M.; software, R.T.; validation, M.M. and R.T.; formal analysis, M.M.; investigation, M.M.; resources, M.I.; data curation, M.I.; writing—original draft preparation, M.M.; writing—review and editing, M.M.; visualization, M.M. and R.T.; supervision, M.M.; project administration, M.I.; funding acquisition, M.I. All authors have read and agreed to the published version of the manuscript. Funding: This research was funded mainly by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) “Integrated Program for Next-generation Volcano Research and Human Resource Development” and partially by the Council for Science, Technology and Innovation (CSTI), “Cross-ministerial Strategic Innovation Promotion Program (SIP)”. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable. Data Availability Statement: Not applicable. Acknowledgments: This work was supported by the MEXT “Integrated Program for Next-generation Volcano Research and Human Resource Development”. The radar data analyzed in the present study were obtained from the Ministry of Land, Infrastructure, Transport and Tourism (MLIT) of Japan using a framework designed by the Data Integration and Analysis System (DIAS), which is funded by MEXT. Conﬂicts of Interest: The authors declare no conﬂict of interest. The funders had no role in the design of the study, the collection, analysis, or interpretation of the data, the writing of the manuscript, or the decision to publish the results. Glossary All symbols and units used in the present study are listed below. Conventional units in radar meteo- rology were used instead of Système international (SI) units. gy y List of symbols and units used in the present study. gy y List of symbols and units used in the present study. 22 of 24 Atmosphere 2021, 12, 601 Symbol Description Unit Ai Effective measured area of the i-th size bin mm2 CA Volcanic ash mass concentration kg m−3 Cij Number of particles measured in i-th diameter bin and j-th Velocity bin - D Particle diameter mm D0 Median volume diameter mm Dm Mass-weighted mean diameter mm Dmax Maximum particle diameter mm Dmin Minimum particle diameter mm ∆Di Bin size of the i-th diameter channel mm N(D) Particle size distribution (PSD) mm−1 m−3 N(Di) Number of particles from Di to Di + ∆Di per unit volume mm−1 m−3 N0 Intercept parameter of gamma PSD mm–1–µ m−3 NT Total number of volcanic ash particles m−3 Nw Normalized intercept parameter of gamma PSD mm−1 m−3 nd Number of diameter bins - nv Number of velocity bins - RA Ash fall rate; 1 (kg m−2h−1) = ρp/ρb (mm h−1) mm h−1 ∆t Sampling time (60 s) s Vj Fall velocity measured at the j-th velocity bin m s−1 Vt Fall velocity of a volcanic ash particle m s−1 Z Equivalent reﬂectivity factor mm6 m−3 Λ Slope parameter of gamma PSD mm−1 µ Shape parameter of gamma PSD - ρb Bulk density of ash deposits (= 1.25 × 103 kg m−3) kg m−3 ρp Density of a solid ash particle (= 2.5 × 103 kg m−3) kg m−3 References 1. 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Introduction: Part 2: Volcanic ash: Generation and spatial variations. In Volcanic Ash Hazard Observation; Mackie, S., Cashman, K., Ricketts, H., Rust, A., Watson, M., Eds.; Elsevier: Amsterdam, The Netherlands, 2016; pp. 5–22. 9. Pioli, L.; Bonadonna, C.; Pistolesi, M. Reliability of total grain-size distribution of tephra deposits. Sci. Rep. 2019, 9, 10006. [CrossRef] 9. Pioli, L.; Bonadonna, C.; Pistolesi, M. Reliability of total grain-size distribution of tephra deposits. Sci. Rep. 2019, 9, 10006. [CrossRef] 10. Suh, S.-H.; Maki, M.; Iguchi, M.; Lee, D.-I.; Yamaji, A.; Momotani, T. Free-fall experiments of volcanic ash particles using a 2-D video disdrometer. Atmos. Meas. Tech. 2019, 12, 5363–5379. [CrossRef] 11. References Drop size distribution comparisons between Parsivel and 2-D video 19. Lanza, L.G.; Vuerich, E. The WMO ﬁeld intercomparison of rain intensity gauges. Atmos. Res. 2009 20. Thurai, M.; Petersen, W.A.; Tokay, A.; Schultz, C.; Gatlin, P. Drop size distribution comparisons between Parsivel and 2-D video disdrometers. Adv. Geosci. 2011, 30, 3–9. [CrossRef] 21. Joss, J.; Waldvogel, A. Ein spektrograph für niederschlagstropfen mit automatischer auswertung (A with automatic interpretation). Pure Appl. Geophys. 1967, 68, 240–246. [CrossRef] 22. Kagoshima Local Meteorological Observatory, 2019. List of Sakurajima Eruptions in 2013. Available online: https://www.jma- net.go.jp/kagoshima/vol/data/skr_exp_list/skr_exp_2019.html (accessed on 23 April 2021). (In Japanese) 23. JMA. Available online: https://www.jma-net.go.jp/kagoshima/vol/data/skr_exp_2013.html (accessed on 3 October 2019). 24 M ki M Ki Y K b i T Hi K L D I I hi M A l f h di i l h d d f l i 23. JMA. Available online: https://www.jma-net.go.jp/kagoshima/vol/data/skr_exp_2013.html (accessed on 3 October 2019). 24. Maki, M.; Kim, Y.; Kobori, T.; Hirano, K.; Lee, D.-I.; Iguchi, M. Analyses of three-dimensional weather radar data from volcanic eruption clouds. J. Volcanol. Geotherm. Res. 2021. [CrossRef] J p // j g jp/ g / / / _ p_ ( 24. Maki, M.; Kim, Y.; Kobori, T.; Hirano, K.; Lee, D.-I.; Iguchi, M. Analyses of three-dimensional weather eruption clouds. J. Volcanol. Geotherm. Res. 2021. [CrossRef] 25. DIAS. XRAIN Precipitation Original Data search and Download System. Available online: https://auth.diasjp.net/cas/login? service=http://xrain.diasjp.net/original/&locale=en (accessed on 13 March 2019). 26. Guyot, A.; Pudashine, J.; Protat, A.; Uijlenhoet, R.; Pauwels, V.R.N.; Seed, A.; Walker, J.P. Effect of disdrometer type on rain drop size distribution characterisation: A new dataset for south-eastern Australia. Hydrol. Earth Syst. Sci. 2019, 23, 4737–4761. [CrossRef] 27. Ulbrich, C.W. Natural variations in the analytical form of the drop size distribution. J. Clim. Appl. Meteorol. 1983, 22, 1764–1775. [CrossRef] 28. Ulbrich, C.W.; Atlas, D. Assessment of the contribution of differential polarization to improved rainfall measurements. Radio Sci. 1984, 19, 49–57. [CrossRef] 29. Chandrasekar, V.; Bringi, V.N. Simulation of radar reﬂectivity and surface measurements of rainfall. J. Atmos. Ocean. Technol. 1987, 4, 464–478. [CrossRef] 30. Marshall, J.S.; Palmer, W.M. The distribution of raindrops with size. J. Meteorol. 1948, 5, 165–166. [C 31. Miwa, T.; Maki, M.; Kozono, T.; Fujita, E.; Tanada, T.; Iguchi, M. Experimental measurement on falling velocity of volcanic ash from Sakurajima volcano by using PARSIVEL disdrometer. Annu. Disas. Prev. Res. Inst. Kyoto Univ. 2015, 58, 91–94. References Kozono, T.; Iguchi, M.; Miwa, T.; Maki, M.; Maesaka, T.; Miki, D. Characteristics of tephra fall from eruptions at Sakurajima volcano, revealed by optical disdrometer measurements. Bull. Volcanol. 2019, 81. [CrossRef] 12. Iguchi, M.; Nakamichi, H.; Tanaka, H.; Ohta, Y.; Shimizu, A.; Miki, D. Int Sakurajima volcano, Japan. J. Disast. Res. 2019, 14, 798–809. [CrossRef] j p 13. Schönhuber, M.; Lammer, G.; Randeu, W.L. The 2D video disdrometer. In Precipitation: Advances in Measurement, Estimation and Prediction; Michaelides, S., Ed.; Springer: Berlin/Heidelberg, Germany, 2008; pp. 3–31. p g g y pp 14. Kruger, A.; Krajewski, W.F. Two-dimensional video disdrometer: A description. J. Atmos. Ocean. Technol. 2002, 19, 602–617. [CrossRef] [ ] 15. Löfﬂer-Mang, M.; Joss, J. An optical disdrometer for measuring size and velocity of 852 hydrometeors. J. Atmos. Ocean. Technol. 2000, 17, 130–139. [CrossRef] 15. Löfﬂer-Mang, M.; Joss, J. An optical disdrometer for measuring size and velocity of 852 hydrometeors. J. Atmos. Ocean. Technol. 2000, 17, 130–139. [CrossRef] 23 of 24 23 of 24 Atmosphere 2021, 12, 601 16. Tokay, A.; Wolff, D.B.; Petersen, W.A. Evaluation of the new version of the laser-optical disdrometer, OTT Parsivel2. J. Atmos. Ocean. Technol. 2014, 31, 1276–1288. [CrossRef] 2 17. OTT. Operating Instructions, Prese Kempten, Germany, 2011; 46p. 17. OTT. Operating Instructions, Present Weather Sensor, OTT Parsivel2; Document Number 70.210.001.B.E 04-0911; Hydromet GmbH: Kempten, Germany, 2011; 46p. Kempten, Germany, 2011; 46p. p y p 18. Krajewski, W.F.; Kruger, A.; Caracciolo, C.; Golé, P.; Barthes, L.; Creutin, J.-D.; Delahaye, J.-Y.; Nikolopoulos, E.I.; Ogden, F.; Vinson, J.-P. DEVEX—Disdrometer evaluation experiment: Basic results and implications for hydrologic studies. Adv. Water Resour. 2006, 29, 311–325. [CrossRef] 19. Lanza, L.G.; Vuerich, E. The WMO ﬁeld intercomparison of rain intensity gauges. Atmos. Res. 2009, 94, 534–543. [CrossRef] 20 Th i M P t W A T k A S h lt C G tli P D i di t ib ti i b t P i l d 2 D id 19. Lanza, L.G.; Vuerich, E. The WMO ﬁeld intercomparison of rain intensity gauges. Atmos. Res. 2009, 94, 534–543. [CrossRef] 20. Thurai, M.; Petersen, W.A.; Tokay, A.; Schultz, C.; Gatlin, P. Drop size distribution comparisons between Parsivel and 2-D video disdrometers. Adv. Geosci. 2011, 30, 3–9. [CrossRef] 19. Lanza, L.G.; Vuerich, E. The WMO ﬁeld intercomparison of rain intensity gauges. Atmos. Res. 2009, 94, 534–543. [CrossRef] 20. Thurai, M.; Petersen, W.A.; Tokay, A.; Schultz, C.; Gatlin, P. References Available online: http://hdl.handle.net/2433/210085 (accessed on 23 April 2021). (In Japanese with English abstract). 32. Kozono, T.; Miwa, T.; Maki, M.; Maesaka, T.; Miki, D.; Iguchi, M. PARSIVEL tephra-fall observations at Sakurajima Volcano. Annu. Disast. Prev. Res. Inst. Kyoto Univ. 2015, 58B, 86–90, (In Japanese with English abstract). Available online: http: //hdl.handle.net/2433/210086 (accessed on 23 April 2021). (In Japanese with English abstract). Annu. Disast. Prev. Res. Inst. Kyoto Univ. 2015, 58B, 86 90, (In Japanese with English abstract). Available online: http: //hdl.handle.net/2433/210086 (accessed on 23 April 2021). (In Japanese with English abstract). //hdl.handle.net/2433/210086 (accessed on 23 April 2021). (In Japanese with English abstract). 34. Harris, D.M.; Rose, W.I. Estimating particle sizes, concentrations, and total mass of ash in volcanic clouds using weather radar. JGR Oceans 2019, 88, 10969–10983. [CrossRef] 35. Sekhon, R.S.; Srivastava, R.C. Doppler radar observations of drop-size distributions in a thunderstorm. J. Atmos. Sci. 1971, 28, 983–994. [CrossRef] 36. Willis, P.T. Functional ﬁts to some observed drop size distributions and parameterization of rain. J. Atmos. Sci. 1984, 41, 1648–1661. [CrossRef] 37. Sempere Torres, D.; Porrà, J.M.; Creutin, J.-D. 1998: A general formulation for raindrop size distri 103, 1785–1797. [CrossRef] 38. Testud, J.; Le Bouar, E.; Obligis, E.; Ali Mehenni, M. The rain proﬁling algorithm applied to polarimetric weather radar. J. Atmos. Ocean. Technol. 2000, 17, 332–356. [CrossRef] 39. Testud, J.; Oury, S.; Black, R.A.; Amayenc, P.; Dou, X. The concept of “normalized” distribution to describe raindrop spectra: A tool for cloud physics and cloud remote sensing. J. Appl. Meteorol. 2001, 40, 1118–1140. [CrossRef] p y g pp 40. Delanoë, J.; Protat, A.; Testud, J.; Bouniol, D.; Heymsﬁeld, A.J.; Bansemer, A.; Brown, P.R.A.; Forbes, R.M. Statistical properties of the normalized ice particle size distribution. J. Geophys. Res. 2005, 110, D10201. [CrossRef] 41. Kozu, T.; Nakamura, K. Rainfall parameter estimation from dual-radar measurements combining reﬂectivity proﬁle and path- integrated attenuation. J. Atmos. Ocean. Technol. 1991, 8, 251–271. [CrossRef] 42. Ulbrich, C.W.; Atlas, D. Rainfall microphysics and radar properties: Analysis methods for drop size spectra. J. Appl. Meteorol. 1998, 37, 912–923. [CrossRef] 42. Ulbrich, C.W.; Atlas, D. Rainfall microphysics and radar properties: Analysis methods for drop size spectra. J. Appl. Meteorol. 1998, 37, 912–923. [CrossRef] 44. Sekhon, R.S.; Srivastava, R.C. Snow size spectra and radar reﬂectivity. J. Atmos. Sci. 1970, 28, 983–994. [CrossRef] 44. Sekhon, R.S.; Srivastava, R.C. Snow size spectra and radar reﬂectivity. J. Atmos. Sci. 1970, 28, 983–994. References [CrossRef] Atmosphere 2021, 12, 601 24 of 24 24 of 24 45. Bagheri, G.; Bonadonna, C. Aerodynamics of volcanic particles: Characterization of size, shape, and settling velocity. In Volcanic Ash Hazard Observation; Mackie, S., Cashman, K., Ricketts, H., Rust, A., Watson, M., Eds.; Elsevier: Amsterdam, The Netherlands, 2016; pp. 39–52. [CrossRef] pp 46. Manzella, I.; Bonadonna, C.; Phillips, J.C.; Monnard, H. The role of gravitational instabilities in depositio 2015, 43, 211–214. [CrossRef] [ ] 47. Freret-Lorgeril, V.; Gilchri, J.; Donnadieu, F.; Jellinek, A.M.; Delanoë, J.; Latchimy, T.; Vinson, J.P.; Caudoux, C.; Peyrin, F.; Hervier, C.; et al. Ash sedimentation by ﬁngering and sediment thermals from wind-affected volcanic plumes. Earth Planet. Sci. Lett. 2020, 534. [CrossRef] 48. Ganser, G.H. A rational approach to drag prediction of spherical and nonspherical particles. 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Soc. Japan 2020, 98, 299–317. [CrossRef] Meteorol. Soc. Japan 2020, 98, 299–317. [CrossRef] p 53. Coata, A.; Folch, A.; Macedonio, G. A model for wet aggregation of ash particles in volcanic plumes and clouds: 1. Theoretical formulation. J. Geophys. Res. 2010, 115, B09201. [CrossRef] 53. Coata, A.; Folch, A.; Macedonio, G. A model for wet aggregation of ash formulation. J. Geophys. Res. 2010, 115, B09201. [CrossRef] p y 54. Folch, A.; Costa, A.; Durant, A.; Macedonio, G. A model for wet aggregation of ash particles in volcanic plumes and clouds: 2. Model application. J. Geophys. Res. 2010, 115, B09202. [CrossRef] 55. Brown, R.J.; Bonadonna, C.; Durant, A.J. A review of volcanic ash aggregation. Phys. Chem. Earth Parts A/B/C 2012, 45–46, 65–78. [CrossRef] 56. Markowitz, A.H. Raindrop size distribution experiments. J. Appl. Meteorol. References 1976, 15, 1029–1166. [Cross 57. Feingold, G.; Levin, Z. 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W3180674549.txt	https://diglib.uibk.ac.at/ulbtirolfodok/content/titleinfo/6306640/full.pdf	de		Die mittelalterlichen Handschriften in der Bibliothek des Augustiner Chorherrenstiftes Neustift	Verlag der österreichischen Akademie der Wissenschaften eBooks	2,021	cc-by	259,780	ÖSTERREICHISCHE AKADEMIE DER WISSENSCHAFTEN PHILOSOPHISCH-HISTORISCHE KLASSE DENKSCHRIFTEN, 529. BAND VERÖFFENTLICHUNGEN ZUM SCHRIFT- UND BUCHWESEN DES MITTELALTERS ISBN 978-3-7001-8625-0 9 783700 STAMPFER · SCHRETTER-PICKER · DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT REIHE IV, BAND 9 186250 Made in Europe Dph. 529 URSULA STAMPFER – CLAUDIA SCHRETTER-PICKER DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT UNTER MITARBEIT VON PETRA AUSSERLECHNER, GIULIA GABRIELLI, PATRIK KENNEL, GABRIELA KOMPATSCHER GUFLER, WALTER NEUHAUSER, ANNA PINTER, SUSANNE RISCHPLER, MARIA STIEGLECKER, LAV ŠUBARIĆ URSULA STAMPFER – CLAUDIA SCHRETTER-PICKER DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT ÖSTERREICHISCHE AKADEMIE DER WISSENSCHAFTEN PHILOSOPHISCH-HISTORISCHE KLASSE DENKSCHRIFTEN, 529. BAND VERÖFFENTLICHUNGEN ZUM SCHRIFT- UND BUCHWESEN DES MITTELALTERS HERAUSGEGEBEN VON WALTER POHL REIHE IV MONOGRAPHIEN HERAUSGEGEBEN VON CHRISTINE GLASSNER UND MARIA THEISEN BAND 9 URSULA STAMPFER – CLAUDIA SCHRETTER-PICKER DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT UNTER MITARBEIT VON PETRA AUSSERLECHNER, GIULIA GABRIELLI, PATRIK KENNEL, GABRIELA KOMPATSCHER GUFLER, WALTER NEUHAUSER, ANNA PINTER, SUSANNE RISCHPLER, MARIA STIEGLECKER, LAV ŠUBARIĆ Angenommen durch die Publikationskommission der philosophisch-historischen Klasse der ÖAW: Michael Alram, Bert G. Fragner, Andre Gingrich, Hermann Hunger, Sigrid Jalkotzy-Deger, Renate Pillinger, Franz Rainer, Oliver Jens Schmitt, Danuta Shanzer, Peter Wiesinger, Waldemar Zacharasiewicz Veröffentlicht mit Unterstützung des Austrian Science Fund (FWF): PUB 751-Z Open Access: Wo nicht anders festgehalten, ist diese Publikation lizenziert unter der Creative Commons Lizenz Namensnennung 4.0 Open access: Except where otherwise noted, this work is licensed under a Creative Commons Attribution 4.0 Unported License. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ Bibliografische Information der Deutschen Bibliothek Die Deutsche Bibliothek verzeichnet diese Publikation in der Deutschen Nationalbibliografie, detaillierte bibliografische Daten sind im Internet über <http://dnb.ddb.de> abrufbar. Diese Publikation wurde einem anonymen, internationalen Begutachtungsverfahren unterzogen. This publication was subject to international and anonymous peer review. Peer review is an essential part of the Austrian Academy of Sciences Press evaluation process. Before any book can be accepted for publication, it is assessed by international specialists and ultimately must be approved by the Austrian Academy of Sciences Publication Committee. Die verwendete Papiersorte in dieser Publikation ist DIN EN ISO 9706 zertifiziert und erfüllt die Voraussetzung für eine dauerhafte Archivierung von schriftlichem Kulturgut. Bestimmte Rechte vorbehalten. ISBN 978-3-7001-8625-0 Copyright © Österreichische Akademie der Wissenschaften, Wien 2021 Satz: Auer Grafik Buch Web, 1050 Wien Druck: Prime Rate, Budapest https://epub.oeaw.ac.at/8625-0 https://verlag.oeaw.ac.at Made in Europe Inhaltsübersicht Vorwort �� 7 TEIL I: EINLEITUNG �� 11 1. DIE STIFTSBIBLIOTHEK VON NEUSTIFT: GESCHICHTE DER BIBLIOTHEK UND DES BESTANDES .......................................................................................................... 13 2. ÜBERSICHT ÜBER DEN HANDSCHRIFTENBESTAND ................................................... 29 2.1 Signaturen �� 2.2 Fachgebiete / Inhalte �� 2.3 Zeitliche Verteilung �� 2.4 Entstehungsorte �� 2.5 Schriften und Schreiberhände �� 2.6 Beschreibstoffe �� 2.7 Sprachliche Verteilung �� 2.8 Provenienzen und Besitzeinträge ............................................................................................. 3. 29 30 31 32 33 33 34 34 BESONDERHEITEN DER ERSCHLIESSUNG UND IHRE ERGEBNISSE ........................ 37 3.1 Wasserzeichen �� 3.2 Illuminierte Handschriften �� 3.3 Musikalien �� 3.4 Einbände ................................................................................................................................... 37 40 44 48 4. BISHERIGE ERSCHLIESSUNG DES HANDSCHRIFTENBESTANDES �� 55 5. ZUR ANLAGE DES KATALOGS �� 57 5.1 Zur Erschließungsmethode ....................................................................................................... 57 5.2 Zur Anlage der Handschriftenbeschreibungen ......................................................................... 58 TEIL II: KATALOGTEIL �� 61 Verzeichnis der beschriebenen Handschriften ............................................................................... Verzeichnis der Abkürzungen und Siglen ...................................................................................... Verzeichnis der abgekürzt zitierten Literatur ................................................................................. 63 69 71 KATALOG �� 79 ANHANG �� 381 Verzeichnis der heute noch an der ULB Tirol befindlichen, zu Beginn des 19. Jahrhunderts aus Neustift dorthin gebrachten Handschriften �� Konkordanz Signaturen der Stiftsbibliothek Neustift – Projektnummern der Hill Monastic Manuscript Library �� Verzeichnis der Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren �� Verzeichnis der Hymnen �� Verzeichnis der Reimoffizien �� Verzeichnis der Sequenzen �� Verzeichnis und Abbildungen der Abreibungen ............................................................................ 383 390 392 398 405 406 411 REGISTER �� 421 Textanfänge (Incipit) ...................................................................................................................... Unvollständige Textanfänge (Incipit mutile) ................................................................................. Verfasser und Werktitel .................................................................................................................. Vorbesitzer, Benützer, Auftraggeber, Widmungsträger und Käufer ............................................... Schreiber, Schreiberverse und -sprüche �� Entstehungszeiten (Datierungen) ................................................................................................... Entstehungsorte .............................................................................................................................. 423 458 461 470 471 472 473 7 Vorwort Die vorliegende Publikation ist das Ergebnis eines von der Autonomen Provinz Bozen-Südtirol, Abteilung Bildungsförderung, Universität und Forschung geförderten Forschungsprojektes (1. Wettbewerbsausschreibung für Projekte im Bereich der wissenschaftlichen Forschung der Autonomen ProvinzBozen-Südtirol). Zwischen 2011 und 2014 konnten in diesem Kooperationsprojekt zwischen der Philosophisch-Theologischen Hochschule Brixen und der Universitäts- und Landesbibliothek Tirol (im Folgenden „ULB Tirol“) in Innsbruck die mittelalterlichen Handschriften in der Bibliothek des Augustiner Chorherrenstiftes Neustift und in der Bibliothek des Priesterseminars in Brixen bearbeitet werden.1 Die geografische Nähe des Klosters Neustift zum Brixner Priesterseminar und die Wurzeln bzw. Entwicklungen der beiden Institutionen bedingen vielfache wechselseitige Beziehungen, die sich nicht zuletzt in ihren Büchersammlungen niederschlagen. Aus diesem Grund schien eine parallele Bearbeitung der beiden Bestände äußerst sinnvoll. So konnten die Bezüge klar herausgearbeitet und wertvolle Synergieeffekte erzielt werden. In den vorliegenden Beschreibungen der Neustifter Handschriften sind daher auch immer wieder Verweise auf die Bestände des Priesterseminars zu finden. Seit der vorübergehenden Auflösung des Stiftes in der Zeit der Zugehörigkeit Tirols zu Bayern (1806–1814) befindet sich ein Teil der Neustifter Handschriften in den Beständen der ULB Tirol. Da sie dort bereits erschlossen wurden2, konnte durch das Projekt eine „virtuelle“ Zusammenführung der heute getrennten Bestände ermöglicht werden. Zum Gelingen des Katalogs haben zahlreiche Personen und Institutionen entscheidend beigetragen: An erster Stelle geht der Dank an die Autonome Provinz Bozen-Südtirol für ihre großzügige finanzielle Förderung. Nicht möglich gewesen wäre das Projekt ohne das Vertrauen von Prälat Prof. Dr. Franz Georg Untergaßmair, Altabt des Stiftes Neustift, der den Bearbeiterinnen und Bearbeitern ungehinderten Zutritt zu den wertvollsten Schätzen der Stiftsbibliothek gewährte und sie im Kloster gastfreundlich aufnahm. Herzlich gedankt sei auch dem ehemaligen Stiftsdechant und Stiftsarchivar Herrn Dr. Theobald Innerhofer (gest. 2012), der das Unternehmen mit großem Interesse und sachkundiger Hilfe begleitete. Ebenso danken wir dem ehemaligen Stiftsbibliothekar Herrn Dipl.-Rpfl. Mag. Rüdiger Weinstrauch für seine vielfältige fachliche und persönliche Unterstützung. Ein Dank gebührt auch dem Prälaten Mag. Eduard Fischnaller (seit 2015), der nach Abschluss der Erschließungsarbeiten die Kosten für die in Zusammenarbeit mit der Universitätsbibliothek Bozen durchgeführte Digitalisierung des Handschriftenbestandes übernahm. Für die Bereitschaft, das Gesamtprojekt zur Erschließung des Bestandes in der Stiftsbibliothek Neustift und in der Priesterseminarbibliothek in Brixen an der Philosophisch-Theologischen Hochschule Brixen organisatorisch ansiedeln zu dürfen, sei besonders dem ehemaligen Regens des Priester1 2 Der Katalog zu den mittelalterlichen Handschriften des Brixner Priesterseminars ist in Vorbereitung. Das seitens des Fonds zur Förderung der wissenschaftlichen Forschung in Österreich (FWF) geförderte Langzeitprojekt zur Erschließung der Handschriften der ULB Tirol in Innsbruck konnte 2017 abgeschlossen werden. Unter der Leitung von Walter Neuhauser (gest. 2016) erschienen seit 1987 insgesamt zehn Teilbände, die auch die ehemaligen Neustifter Bestände mitberücksichtigen; vgl. Anm. 100. 8 Vorwort seminars und seit 2011 Bischof der Diözese Bozen-Brixen, Prof. Dr. Ivo Muser, den Bibliotheks direktoren, Prof. Dr. Arnold Stiglmair und seinem Nachfolger Prof. Dr. Ulrich Fistill, sowie dem Verwalter Dr. Thomas Schraffl gedankt. Dank gilt auch Mag. Claudia Kaser und ihrem Team der Biblio thekder Philosophisch-Theologischen Hochschule für die Möglichkeit, dort arbeiten und recherchieren zu dürfen. Gedankt sei der Leopold-Franzens-Universität Innsbruck und dem ehemaligen Rektor Univ.-Prof. Dr. Karlheinz Töchterle, der diesem Projektunternehmen von Anfang an positiv gegenüberstand und so die Funktion der ULB Tirol als Kooperationspartner ermöglichte. Den Mitarbeitenden in der Verwaltung beider Institutionen ein herzliches Danke für ihre Be gleitung bei der Projektanbahnung und -abwicklung. Die ULB Tirol mit ihrem früheren Direktor HR Dr. Martin Wieser (bis 2014) begrüßte nicht nur die Initiative zur Erschließung dieses Komplementärbestandes zu den in Innsbruck verwahrten Handschriften Neustifter Provenienz. Er stellte dem Projekt auch über die Projektmittel hinausgehende personelle, infrastrukturelle und finanzielle Ressourcen zur Verfügung. Ihm und seinen Nachfolgerinnen im Amt, Dr. Elisabeth Frasnelli (2014–2015) und HR Mag. Eva Ramminger (seit 2016), sowie dem interimistischen Leiter HR Dr. Klaus Niedermair (2015–2016), sei für ihr stetes Wohlwollen und ihre Unterstützung gedankt. Der Abteilung für Sondersammlungen mit ihrem Abteilungsleiter OR Mag. Peter Zerlauth gebührt nicht nur für die Bereitstellung der dortigen Infrastruktur und die Schaffung idealer Arbeitsbedingungen, sondern auch für mannigfache Hilfestellungen und das stete Entgegenkommen ein großer Dank, ebenso dem ehemaligen Mitarbeiter Michael Springer für seine Unterstützung. Ein abschließender Dank gilt der Direktorin der Universitätsbibliothek Bozen, Mag. Gerda Winkler, die die Digitalisierung der Neustifter Bestände unter der Ägide ihrer Mitarbeiterin Mag. Dr. Ursula Stampfer (2017–2020) begrüßte und förderte. Die Projektleitung oblag Mag. Dr. Ursula Stampfer, die hierfür mit Projektmitteln von der Philo sophisch-Theologischen Hochschule in Brixen angestellt wurde. Für die ULB Tirol als Projektpartner wurde MMag. Dr. Claudia Schretter-Picker im Rahmen ihrer Tätigkeit an der dortigen Abteilung für Sondersammlungen die Verantwortung übertragen. Beide übernahmen neben der Projektadministration und Qualitätssicherung auch die Beschreibung von Handschriften sowie die Redaktion bzw. die Vorbereitung der Drucklegung. An der Beschreibung der Handschriften arbeiteten mehrere Fachkräfte mit unterschiedlichem Beschäftigungsausmaß und unterschiedlicher Anstellungsdauer mit, angesiedelt je weils an der Abteilung für Sondersammlungen der ULB Tirol (in alphabetischer Reihenfolge): Mag. Petra Ausserlechner (2011, philosophisch-theologische Handschriften), Mag. Patrik Kennel (2013–2014, historisch-juristische Handschriften), MMag. Anna Pinter (2013–2014, philosophisch-theologische sowie philologische Handschriften, Redaktion). Die kunsthistorische Analyse und Auswertung übernahm Dr. Susanne Rischpler im Rahmen eines Werkvertrags (2012–2014). Mit der Erstellung der Register wurde MMag. Sissy Mederle im Rahmen eines Werkvertrages (2013–2014) betraut, die Überarbeitung und Vereinheitlichung wurde durch die Projektleiterinnen vorgenommen. Die Erstellung der Anhänge sowie die Durchreibungen der Einbände besorgte Mag. Dr. Ursula Stampfer, fallweise mit Unterstützung von PraktikantInnen. Ein Dank geht an Gionata Brusa MA und Mag. Vera-Maria Kathrein für die kritische Durchsicht der liturgischen Handschriften. Zurückgegriffen werden konnte auch auf die Unterstützung „von außen“, nämlich von Spezialisten, die bei der Bearbeitung ohne Kosten für das Projektbudget aktiv mitwirkten. Ihnen sei herzlichst gedankt: Ao. Univ.-Prof. Mag. Dr. Gabriela Kompatscher Gufler (philologische Handschriften) sowie Ass.-Prof. Mag. Dr. Lav Šubarić (naturwissenschaftliche Handschriften) vom Institut für Klassische Philologie und Neulateinische Studien der Universität Innsbruck bzw. vom Ludwig-Boltzmann-Institut für Neulateinische Studien in Innsbruck. Dr. Giulia Gabrielli (Freie Universität Bozen, Fakultät für Bildungswissenschaften Brixen) widmete sich musikwissenschaftlichen Fragen. Die betaradiografischen Aufnahmen und die Bestimmung der Wasserzeichen besorgte Mag. Dr. Maria Stieglecker MAS im Vorwort 9 Rahmen ihrer Tätigkeit an der Abteilung Schrift- und Buchwesen des Mittelalters am Institut für Mittelalterforschung der Österreichischen Akademie der Wissenschaften (im Folgenden „ÖAW“). Großer Dank gebührt HR Mag. Dr. Walter Neuhauser (gest. 2016), früherer Direktor der ULB Tirol und ehemaliger Leiter des 2017 abgeschlossenen Langzeitprojektes zur Erschließung der Handschriften der ULB Tirol. Mit Interesse verfolgte und förderte er den Fortgang der Arbeit und gab im Hinblick auf konkrete Fragen zum Handschriftenbestand der Stiftsbibliothek Neustift und zur Kodikologie im Allgemeinen wichtige Anregungen und unverzichtbare Hinweise. Darüber hinaus brachte er sich in die praktische Erschließungsarbeit vor Ort und an der Abteilung für Sondersammlungen durch die Übernahme von Beschreibungen maßgeblich ein. Ihm sei auch für die kritische Durchsicht der Katalogisate herzlich gedankt. Bei fachspezifischen Einzelfragen konnten die Bearbeiterinnen und Bearbeiter auf die Hilfe zahlreicher Kolleginnen und Kollegen sowie Experten (vgl. entsprechende Hinweise bei den einzelnen Katalogisaten) zählen, wofür wir ihnen sehr zu Dank verpflichtet sind. Zudem bedanken wir uns bei den Verantwortlichen der Handschriftenabteilungen anderer Bibliotheken, insbesondere der Kathedralbibliothek der Erzdiözese Esztergom-Budapest, der Österreichischen Nationalbibliothek (im Folgenden „ÖNB“) in Wien und der Bayerischen Staatsbibliothek (im Folgenden „BSB“) in München, die durch die Gewährung guter Arbeitsbedingungen den Fortgang der Katalog isierung förderten. Für die Durchsicht des Manuskriptes und wertvolle Anregungen und Hinweise im Rahmen eines Werkvertrages (2014) ist Dr. Alois Haidinger und Dr. Franz Lackner herzlich zu danken. Für die Letztdurchsicht des Manuskripts im Rahmen ihrer Tätigkeit an der Abteilung für Sondersammlungen der ULB Tirol gilt MMag. Magdalena Rufin Dank. Ein Dank gebührt MMag. Anna Pinter für das umsichtige Lektorat. Die erhobenen Daten wurden bereits während der Erschließung über das von Dr. Alois Haidinger entwickelte und von DI Emanuel Wenger und Mag. Dr. Christine Glaßner betreute Handschriftenportal www.manuscripta.at zugänglich gemacht. Ihnen sei für die gute Zusammenarbeit und das stete Ent gegenkommen herzlich gedankt. Im Sinne des Open Access sind die Daten des gedruckten Katalogs über dieses Portal abrufbar. Ebendort werden auch die dank der großzügigen Finanzierung seitens des Klosters Neustift von den beiden Werkvertragsnehmerinnen Edith Kapeller BA MA MA und Mag. Julia Schön MA erstellten Digitalisate zur Verfügung gestellt. Für Unterstützung bei der Abwicklung der Digitalisierung ist dem Bibliothekar der Klosterneuburger Stiftsbibliothek MMag. Dr. Martin Haltrich zu danken. In Zusammenarbeit zwischen der Universitätsbibliothek Bozen und der Universität Venedig (Prof. Paolo Eleuteri) ist auch eine Dateneingabe in die italienische Datenbank www.nuovabibliotecamanoscritta.it geplant. Besonderer Dank gilt der ÖAW, insbesondere dem Direktor des Instituts für Mittelalterforschung, Univ.-Prof. Mag. Dr. Walter Pohl, der Leiterin der Abteilung für Schrift- und Buchwesen des Mittel alters, Mag. Dr. Christine Glaßner, dem Obmann der Publikationskommission, Univ.-Prof. Dr. Hermann Hunger, sowie dem Verlag unter der Leitung von Dipl.-Medienberater Thomas Jentzsch für die Aufnahme dieser Publikation in die Reihe der „Denkschriften“. Ursula Stampfer und Claudia Schretter-Picker Völs am Schlern/Innsbruck, im Frühjahr 2021 TEIL I: EINLEITUNG 13 1. Die Stiftsbibliothek von Neustift Geschichte der Bibliothek und des Bestandes Ursula Stampfer „Neustift besitzt eine große Kirche mit kostbarem Ornat und eine gute Bücherei.“ So charakterisiert der Augsburger Dominikaner Felix Faber 1483 das Augustiner Chorherrenstift Neustift bei Brixen, als er auf seinem Weg ins Heilige Land dort Halt macht.3 Auch der Besucher von heute ist von der licht durchfluteten und mit Fresken reich ausgestatteten Kirche sowie vom großen Bibliothekssaal tief beeindruckt. In der Tat stellt die Neustifter Bibliothek seit dem späten Mittelalter die bedeutendste Tiroler Klosterbibliothek dar, und dies, obwohl sie im Zuge der Bauernkriege, aufgrund mehrerer Brände und durch die vorübergehende Klosteraufhebung zu Beginn des 19. Jahrhunderts große Verluste erfahren hat. Angesichts dieser Bedeutung der Büchersammlung mag es erstaunen, dass bis ins 18. Jahrhundert kaum schriftliche Quellen – weder zum Bestand selbst noch zum Ort, wo die Bücher aufbewahrt wurden – vorliegen und der reiche Schatz an mittelalterlichen Handschriften bis dato noch keiner systematischen wissenschaftlichen Erschließung unterzogen wurde. Als Hartmann, Propst des Augustiner Chorherrenstiftes Klosterneuburg bei Wien, 1140 zum ischof von Brixen geweiht wurde, sollte er die Reformbestrebungen seines Vorgängers Reginbert fort B setzen und dem Brixner Klerus zu neuem Aufschwung verhelfen. Allerdings verpflichtete er nicht die Domherren selbst zu einem Leben nach der Augustinusregel, sondern errichtete mit Unterstützung des Ministerialen Reginbert von Säben und dessen Frau Christina am Kreuzungspunkt zweier wichtiger Reise- und Pilgerrouten, nur wenige Kilometer von seinem Bischofssitz entfernt, ein neues Kloster: Neustift.4 Bücher zählten zweifelsohne zur Grundausstattung des Klosters: Für die Feier der Liturgie, das Gebet sowie die Verwaltung war eine Büchersammlung im Haus unabdingbar. Über die Anfänge der Bibliothek sind wir allerdings nicht unterrichtet. Es ist wohl anzunehmen, dass bereits mit den ersten 3 4 J. Garber, Die Reisen des Felix Faber durch Tirol in den Jahren 1483 und 1484 (Schlern-Schriften 3). Innsbruck 1923, 9. Vgl. C. D. Hassler (Hrsg.), Fratris Felicis Fabri Evagatorium in Terrae Sanctae, Arabiae et Aegipti peregrinationem I. Stuttgardiae 1843, 70: „Est enim monasterium valde solemne et multum abundans […]. Ecclesiam habet magnam cum pretioso ornatu, et librariam bonam“. Zur Geschichte des Stiftes siehe u. a. Th. (H.) Innerhofer, Das Augustiner-Chorherrenstift Neustift, in: H. Obermair, K. Brandstätter, E. Curcel (Hrsg.), Dom- und Kollegiatstifte in der Region Tirol – Südtirol – Trentino in Mittelalter und Neuzeit. Collegialità ecclesiastica nella regione trentino-tirolese dal medioevo all’età moderna (Schlern-Schriften 329). Innsbruck 2006, 223–238 (mit weiterführender Literatur); 850 Jahre Augustiner Chorherrenstift Neustift, hrsg. vom Augustiner Chorherrenstift Neustift. Brixen 1992; Th. (H.) Innerhofer (Red.), 850 Jahre Chorherrenstift Neustift. 1. Südtiroler Landesausstellung. Stift Neustift 30. Mai bis 31. Oktober 1992. Katalog. Brixen 1992; Ders., Die Grundherrschaft des Chorherrenstiftes Neustift bis um 1500. 2 Bde. Diss. Innsbruck 1971; J. Huber (Hrsg.), Aus der Chronik des Stiftes Neustift bei Brixen. Neustift 1956; M. Schrott, Aus der Baugeschichte des Stiftes, in: A. Sparber, M. Schrott (Hrsg.), Aus der Neustifter Klosterchronik. Brixen 1930, 13–59; A. Sparber, Abriß der Geschichte des Chorherrenstiftes Neustift bei Brixen. Brixen 1920. 14 Die Stiftsbibliothek von Neustift Augustiner Chorherren, die von Klosterneuburg nach Neustift übersiedelten, Bücher ins neue Stift gelangten. Zudem dürften sowohl der Gründer als auch die Stifter selbst die Klostergemeinschaft mit Büchern ausgestattet haben. Mit dem Ausbau der Klosteranlage sowie der Gründung einer Schule, zu nächst nur für den stiftsinternen Nachwuchs gedacht, doch schon bald auch für andere Kinder zugäng lich5, waren weitere Bücher notwendig. Vielfach von den Chorherren selbst geschrieben, teilweise käuflich erworben bzw. bei Lohn schreibern in Auftrag gegeben, teilweise als Geschenke dem Stift überlassen, nahm die Anzahl der Bücher sicherlich rasch zu.6 So übergab Berthold, Hospitaler am Heilig-Kreuz-Spital im nahegelege nen Brixen, Propst Nikolaus I. (1346–1359) und dem Konvent von Neustift am dritten Fastensonntag 1353 (24. Februar) vier Bücher mit kanonistischem Inhalt: Primo Decretum cum glosa Bartholomei ordinaria. Item Decretales sextum librum et Clementinas cum suo apparatu. Item Summam Gantfredi et librum feodorum.7 Im Gegenzug erhielt er Begräbnisrecht im Stift, zudem sollte sein Jahrtag wie der eines Konventualen begangen werden. 1383 wurde Hainricus de Uberwisen, plebanus in Lawant, ein Jahrtag sicut unius prelati in oratorio et refectorio als Dank für drei Bücher zugesichert.8 Mit dem zunehmenden wirtschaftlichen Erstarken des Stiftes ging im 15. Jahrhundert auch eine kulturelle Blütezeit einher. Mehr als 500 Höfe, vorwiegend im Pustertal und im Eisacktal gelegen, zinsten dem Stift9 und ermöglichten bauliche Änderungen und die Anschaffung von bedeutenden Kunstwerken. Zahlreiche Altäre, Tafelbilder und Fresken von namhaften Künstlern wie Michael und Friedrich Pacher, Leonhard von Brixen und Marx Reichlich, aber auch die reich mit Miniaturen geschmückten Handschriften aus dem stiftseigenen Skriptorium legen hiervon Zeugnis ab. Eine Schreibstube hat wohl seit jeher im Stift bestanden10, aber erst im 15. Jahrhundert sollte sich ihr guter Ruf verbreiten. 5 6 7 8 9 10 Die Anfänge der Schule in Neustift liegen im Dunkeln, als eigentlicher Gründer der Klosterschule gilt gemeinhin Propst Konrad von Rodank (1178–1200), der vorher als scholasticus am Brixner Dom wirkte. Doch bereits um 1150 übertrug ein Adeliger aus Latzfons dem Stift Güter in Lajen als Dank für die Ausbildung und Ordensauf nahme seines Sohnes. 1281 übergab der Adelige Rupert Maulrapp aus Kastelruth dem Stift einen Hof für die Ausbildung seiner beiden Söhne; nur einer trat anschließend in den Orden ein. Vgl. hierzu Th. (H.) Innerhofer, M. Peintner, E. Delmonego, H. Torggler, Neustift. Von der Klosterschule zum Schülerheim. Brixen 2007, 6–7. Vgl hierzu auch B. Kieferl, Die Stiftsbibliothek in Neustift. Geschichtliche Notizen, zur Feier der dortigen Abtweihe. Brixener Chronik Jg. 26 Nr. 10 (25. Jänner 1913) 2–3, Nr. 13 (1. Februar 1913) 1–2, Nr. 24 (27. Februar 1913) 1–3, Nr. 30 (13. März 1913) 1–2; Jg. 27 Nr. 24 (26. Februar 1914) 1–2. Der einstige Stiftsbibliothekar veröffentlichte in vier Teilen eine sehr anschauliche und lebendige Darstellung der Geschichte der Stiftsbibliothek. Dieser Darstellung folgt Waltraud Lamprecht im ersten Teil ihrer Diplomarbeit über das Historische Werden der Stiftsbibliothek von Neustift, vgl. W. Lamprecht, Die Stiftsbibliothek von Neustift mit besonderer Berücksichtigung der Sektion der deutschen Literatur, Tesi di laurea. Padova, Verona 1980–1981, 6–36. Neustift, Stiftsarchiv, WW 59; vgl. auch Th. Mairhofer, Urkundenbuch des Augustiner-Chorherrenstiftes Neustift in Tirol (Fontes Rerum Austriacarum / Österreichische Geschichts-Quellen II, 34). Wien 1871, 264–265.Drei dieser genannten Handschriften befinden sich bis heute in Neustift: Cod. 7, 99 und 253. Neustift, Stiftsarchiv, WW 66; vgl. auch Mairhofer, Urkundenbuch (wie Anm. 7) 372. In dieser auf den 15. Dezember 1383 datierten Urkunde werden folgende Titel genannt: Summam confessorum maiorem in sollempni volumine; item abreviata libri Sentenciarum cum glossa et postillas perennes de tempore per totum annum. Eine eindeutige Identifizierung der genannten Titel im heute erhaltenen Handschriftenbestand ist nicht möglich. Der erste Eintrag dürfte wohl als „Summa de poenitentia“ oder „Summa confessorum“ des Raimundus de Pennaforte zu identifzieren sein (möglicherweise T. I von Cod. 614). Innerhofer, Grundherrschaft (wie Anm. 4). Zu den Anfängen des stiftseigenen Skriptoriums liegen keine Quellen vor. In der Debatte über den genauen Entstehungsort des berühmten Codex Buranus (München, BSB, Clm 4660) wird seit geraumer Zeit auch immer wieder das Kloster Neustift angeführt. Zuletzt war insbesondere Peter Godman (gest. 2018) davon überzeugt, dass die Liedersammlung um 1230 im Augustiner Chorherrenstift Neustift aufgeschrieben wurde. Vgl. hierzu u. a. G. Steer, „Carmina Burana“ in Südtirol. Zur Herkunft des clm 4660. Zeitschrift für deutsches Altertum und deutsche Literatur 112/1 (1983) 1–37; J. Drumbl, Studien zum Codex Buranus. Aevum 77/2 (2003) 323–356; P. Godman, Rethinking the Carmina Burana (I): The Medieval Context and Modern Reception of the Codex Buranus. Journal of Medieval and Early Modern Studies 45/2 (2015) 245–286; Ders., Die Stiftsbibliothek von Neustift 15 In diese Zeit fällt auch die erste auf uns gekommene Information über einen Bibliotheksraum: Propst Ulrich II. Weingartner (1427–1439) ließ im Jahr 1431 einen „doppelten Bibliotheksraum“ errichten11, der wohl einerseits als Aufbewahrungsort für die Bücher, andererseits als Archiv dienen sollte.12Über den genauen Standort und die Größe des Baus sind wir nicht unterrichtet. Im Juli 1438 reiste der Augustiner Chorherr Friedrich Zollner aus dem Stift Langenzenn bei Nürn berg13, mit dem Neustift seit 1421 geistlich verbrüdert war14, nach Neustift. Zur Förderung des geist lichen Lebens (propter spiritualis vite maiorem fructum ut prefatum est cupiensque saluti anime sue limpidius providere indulsi) hatte Propst Petrus von Langenzenn gemeinsam mit dem Rat seiner Kapi tularen beschlossen, ihn in ein Augustiner Chorherrenstift seiner Wahl (ad locum regularem sibi deputatum) ziehen zu lassen.15 Warum die Wahl Zollners ausgerechnet auf jenes südlich des Brenners fiel, lässt sich heute lediglich vermuten. Sie war aber mit Sicherheit nicht dem Zufall überlassen – nicht umsonst war die Gebetsbrüderschaft mit Neustift im Juni desselben Jahres erneuert worden.16 Ein Grund mag möglicherweise ein vorbildhaftes religiöses Leben der Neustifter Chorherren gewesen sein17; als hochbegabter Schreiber hatte Zollner zudem wohl auch von dem blühenden kulturellen Leben in Neustift und von der dortigen Schreibstube gehört18 und wollte sich vielleicht ebendort weiter bilden bzw. einbringen. Zollner sollte schließlich dem Neustifter Skriptorium ein bleibendes Denkmal setzen: Im Auftrag des Propstes Nikolaus III. Scheiber (1439–1449) schrieb er zwei umfangreiche Gradualbände in Großfolio-Format. Auf etwa 600 Blatt hielt er Text und Melodie sämtlicher Wechsel gesänge der Messen im Laufe eines Kirchenjahres fest, wobei der erste Band dem Temporale und der zweite Band dem Sanctorale vorbehalten ist. Die Rubriken wurden wohl vom Schreiber und somit von Friedrich Zollner selbst ausgeführt, die Fleuronnée-Initialen und der reiche dekorative und figürliche Buchschmuck, der sich durch große Detailfreude auszeichnet, stammen von mehreren Buchmalern (nähere Details siehe Beitrag von Susanne Rischpler). Der üppige Einsatz von Blattgold verlieh der liturgischen Handschrift den bis heute faszinierenden festlichen Charakter. Dieses aufwändige Werk entstand in den Jahren 1439 bis 144619 und somit in einer Zeit, als sich – zumindest zeitweise – auch Oswald von Wolkenstein in Neustift aufhielt. Er war im November 1411 wegen seiner und seiner Familie Verdienste als Pfründner im Stift aufgenommen worden und hatte lebens 11 12 13 14 15 16 17 18 19 Rethinking the Carmina Burana (II). The Child, the Jew, and the Drama. Viator. Medieval and Renaissance Studies 47/1 (2016) 107–122. A. Sparber, Abriß (wie Anm. 4) 43. Dies wird nicht zuletzt dadurch deutlich, dass in jener Zeit das Archiv neu geordnet wurde und die Urkunden mit Signaturen und Kurzregesten versehen wurden. H. Miekisch, Das Augustinerchorherrenstift Neunkirchen am Brand. Seine Geschichte und seine Bedeutung für die Verbreitung der Raudnitzer Reform, Diss. Bamberg 2005; S. Habel, Das Augustiner-Chorherrenstift in Langenzenn von 1409 bis 1533. Horb a. N. 2004. Habel, Langenzenn (wie Anm. 13) 59. Die in Neustift erhalten gebliebenen Rotula (insgesamt fünf, davon lediglich zwei vollständig) belegen, dass Langenzenn regelmäßig von Neustifter Rotelboten aufgesucht wurde (vgl. Rotula aus den Jahren 1515, 1516/17, 1525, siehe Neustift, Stiftsarchiv, XX 58.5). Neustift, Stiftsarchiv, XX 21.1. Das Schreiben des Propstes Petrus von Langenzenn datiert auf den 13. Juli 1438. Neustift, Stiftsarchiv, XX 21 (1438 Juni 19). Im Gegensatz zu vielen anderen Klöstern gab das religiöse Leben im Augustiner Chorherrenstift Neustift Mitte des 15. Jahrhunderts Kardinal Nikolaus von Kues offensichtlich keinen Anlass zur Kritik, im Gegensatz: Er be auftragte den damaligen Propst Kaspar Aigner (1449–1467), dem er erstmals wohl beim Konzil in Basel begeg net sein dürfte, mit der Visitation mehrerer Klöster; vgl. H. Hallauer, Nikolaus von Kues und das Chorherren stift Neustift, in: L. Carlen, F. Steinegger (Hrsg.), Festschrift Nikolaus Grass zum 60. Geburtstag dargebracht von Fachgenossen, Freunden und Schülern, Bd. 1. Innsbruck, München 1974, 309–324, hier insbes. 309–310. Dies kann wohl als Indiz für die dank des Rotelbotensystems bestens funktionierende Kommunikation zwischen den Klöstern gesehen werden; vgl. E. P. Pallhuber, Der Neustifter Rotelbote, in: 850 Jahre Augustiner Chorherrenstift Neustift (wie Anm. 4) 104–111; M. Schrott, Die Conföderationen Neustifts mit anderen Klöstern. Unum Congregati. Mitteilungen der Österreichischen Chorherrenkongregation 4 (1958) 127–158. Im Jahr 1439 wurde Nikolaus Scheiber zum Propst gewählt, der im Kolophon als Auftraggeber genannt wird; 1446 verstarb Friedrich Zollner. Der erste Gradualband wurde im Jahr 1442 abgeschlossen, vgl. Graduale Neocellense I, Bl. 325r–v. 16 Die Stiftsbibliothek von Neustift langes Wohnrecht in einem Haus in unmittelbarer Nähe zur Margarethenkirche sowie Begräbnisrecht er halten.20 Während er zunächst nur kurze Rastzeiten als Durchreisender in Neustift eingelegt zu haben scheint, dürfte er sich ab Mitte der 1430er Jahre bis zu seinem Tod am 2. August 1445 wiederholt und länger dort aufgehalten haben. Die Frage, ob die beiden großformatigen Pergamenthandschriften, in denen er seine Lieder aufzeichnen ließ – die ältere (Hs. A) ist auf 142521, die jüngere (Hs. B) auf 143222 datiert –, im Neustifter Skriptorium entstanden bzw. in irgendeinem Zusammenhang mit diesem zu sehen sind, wird immer wieder aufgeworfen, konnte aber bislang noch nicht eindeutig beantwortet werden.23 Einen anschaulichen Einblick in die Neustifter Schreibstube in ebendieser Blütezeit bietet ein Rezept zur Herstellung von Tinte24, überliefert am Ende einer Handschrift über die Fassmessung aus dem Jahr 1443:25 Item zu ainer mass heffenwassers vier lot gallis, das zerstossent / klain und wann er wol gestossen sey so nempt das heffenwasser un[d] / giessent es daran und lannd das stan funf tag und wann die funf / tag vergangen sind so siedent es dann ain viertail ainer stund / und weil es dann warm ist, so nempt V lot galitzenstain, daz / zerstossent ze mel und tund es darein und ain lot gumi, daz / zerstossent auch und virvent es dar ein, die weil es warm sey / und lannd die dintten stan vier tag. So ist sy gut und schwartz.26 Nur wenige Jahre später sollte der Buchdruck die Herstellung und Verbreitung von Büchern revolutionieren und die Neustifter Bibliothek stark verändern. Wohl dank der verkehrstechnisch äu ßerst günstigen Lage, aber nicht zuletzt auch dank der guten Vernetzung mit anderen Klöstern im süddeutschen Raum27, die sicherlich bei der Beschaffung von Informationen über Neuerscheinungen hilfreich war, konnte Neustift bereits in der zweiten Hälfte des 15. Jahrhunderts eine Vielzahl an gedruckten Büchern erwerben: Nicht weniger als 820 Inkunabeln waren Ende des 18. Jahrhunderts in der Stiftsbibliothek vorhanden.28 Rechnungen über Bücherkäufe sind leider kaum auf uns gekommen, 20 21 22 23 24 25 26 27 28 A. Schwob, „… er ezze mit vns obgenantem Brobst Niklasen … auz vnser schussel“. Beziehungen Oswalds von Wolkenstein zum Propst und Konvent von Neustift, in: 850 Jahre Augustiner Chorherrenstift Neustift (wie Anm. 4) 252–266. Wien, ÖNB, Cod. Vind. 2777. Innsbruck, ULB Tirol, ohne Sign. (Wolkenstein-Hs.); vgl. Katalog ULB Tirol I (wie Anm. 100) 47–50. Vgl. hierzu u. a. W. Neuhauser, Wissenschaftspflege in Neustift im Mittelalter im Spiegel der Hand schriften, in: Ders. (Hrsg.), Beiträge zur Handschriftenkunde und mittelalterlichen Bibliotheksgeschich te. Referate der 7. Tagung österreichischer Handschriftenbearbeiter in Innsbruck/Neustift (Südtirol), Juni 1979 (Innsbrucker Beiträge zur Kulturwissenschaft 47). Innsbruck 1980, 78–79. Unter www.manuscripta. at (Innsbruck, ULB Tirol, Cod. s.n.) ist eine ausführliche aktuelle Bibliografie zur Handschrift B des Oswald von Wolkenstein zu finden, in der Digitalen Bibliothek der ULB Tirol (https://diglib.uibk.ac.at/ulbtirolhds/ content/titleinfo/5914060) ein Volldigitalisat. Eine Auswahlbibliografie zu Tinten- und Farbrezepten des späten Mittelalters ist unter https://mittelalter. hypotheses.org/11576 zu finden (wie alle anderen im Folgenden angeführten Links im April 2021 geprüft). Innsbruck, ULB Tirol, Cod. 593. Entstanden wohl im Wiener Raum, dürfte die Handschrift schon bald Eingang in die Neustifter Bibliothek gefunden haben, vgl. hierzu Katalog ULB Tirol VI (wie Anm. 100) 366–368; K. Girstmair, U. Stampfer, Rätselhafte mathematische Tabellen – der Innsbrucker Codex 593. Codices manuscripti 78/79 (2011) 11–22; G. Storeck, Zahlentafeln in der Fassmessung des 15. Jahrhunderts, in: R. Gebhardt (Hrsg.), Arithmetik, Geometrie und Algebra der frühen Neuzeit. Tagungsband zum wissenschaftlichen Kolloquium „Arithmetik, Geometrie und Algebra der frühen Neuzeit“ vom 11.–13. April 2014 in der Berg- und Adam-Ries Stadt Annaberg-Buchholz (Schriften des Adam-Ries-Bundes AnnabergBuchholz 23). Annaberg-Buchholz 2014, 129–143. Innsbruck, ULB Tirol, Cod. 593, Bl. 23v. Laut den erhalten gebliebenen Rotula war Kloster Neustift zu Beginn des 16. Jahrhunderts mit 91 Klöstern und Stiften in Gebetsbrüderschaft verbunden, vgl. Anm. 18. F. X. Grass, Verzeichnis typographischer Denkmäler aus dem fünfzehenten Jahrhundert, welche sich in der Bibliothek des regulirten Korherrenstiftes des heil. Augustin zu Neustift in Tyrol befinden. Brixen 1798. Vgl. auch G. Manzini, Gli incunaboli della Biblioteca del Convento di Novacella secondo il Catalogo Settecentesco di F. S. Grass. Archivio per l’Alto Adige 47 (1953) 299–317; F. Morandini, Studi sugli incunaboli della biblioteca di Novacella. Archivio per l’Alto Adige 40 (1945) 324–345. Die Inkunabeln wurden 1809 nach Innsbruck gebracht und werden bis heute mehrheitlich dort aufbewahrt; heute befinden sich in der Bibliothek in Neustift nur mehr 33 Wiegendrucke. Die Stiftsbibliothek von Neustift 17 lediglich für den Erwerb einiger Bücher unter Propst Leonhard Pacher (1467–1482) ist eine Notiz mit dem Titel Exposita per Leonhardum prepositum pro libris ornamentis et clenodiis29 erhalten.30 Neben diesem käuflichen Erwerb von Büchern wurden aber bestimmte Texte weiterhin in der stiftseigenen Schreibstube von Hand geschrieben. Spezielle, an den lokalen Ritus und persönliche Bedürfnisse angepasste Mess- und Gebetbücher mussten auch im 16. Jahrhundert händisch hergestellt werden. So enthält Cod. 405, ein um 1501 entstandenes Brevier, das großteils mit dem 1482 in Venedig gedruckten Breviarium Salisburgense31 übereinstimmt, zusätzlich Suffragien von Neustift und von dem Augustiner Chorherrenstift Waldsee.32 In beiden Klöstern war Lukas Härber als Propst tätig.33 Einem gedruckten Brevier nach Salzburger Ritus wurde auf Veranlassung des Brixner Bischofs Georg Golser 1487 ein Anhang beigefügt, damit es in Neustift verwendet werden konnte (Innsbruck, ULB Tirol, Ink. 156 F 20). Ergänzt wurden einige Feste im Sanctorale (u. a. die Feste Ingenuin und Albuin, Monika, Kassian), Angaben zu liturgischen Besonderheiten für Neustift (u. a. Abänderungen von Festrängen bei bestimmten Heiligen) sowie spezielle Suffragien.34 Weiterhin handschriftlich festgehalten wurden auch vor Ort entstandene, oft persönliche Sammlungen von Texten bzw. Kompilationen von Textausschnit ten. Als Beispiele seien hier das 1509 datierte knappe Lehrbuch der Kosmografie mit anschließenden Exzerpten aus antiken Autoren (Innsbruck, ULB Tirol, Cod. 1015), das man möglicherweise für den Schulunterricht benötigte, sowie die von Propst Hieronymus Piesendorfer (1542–1561) 1535/36 verfasste Sermonessammlung (Innsbruck, ULB Tirol, Cod. 1031) genannt. Als besonders begabter Schreiber jener Zeit ragt der Chorherr Stefan Stet(t)ner (gest. 12. Dezem ber 1526)35 heraus. Neben seinem bekanntesten Werk, einem Missale, das er 1524 für bzw. im Auftrag von Propst Augustin Posch (1519–1527) geschrieben hat, sind ein Neustifter Rituale aus dem Jahr 1507 (Cod. 194) und ein Sermo (Innsbruck, ULB Tirol, Cod. 718, dat. 1510) erhalten geblieben. Mit großer 29 30 31 32 33 34 35 Neustift, Stiftsarchiv, Cod. 19, Bl. 190r. Insgesamt sind 19 Werke aufgelistet, die ein breites Interessensspektrum erkennen lassen. Schriften der Kirchenväter Augustinus (De civitate Dei) und Laktanz sind ebenso genannt wie die beiden Hauptwerke des römischen Geschichtsschreibers Flavius Iosephus (Antiquitates Iudaices und De bello Iudaico) und das pflanzenkundliche Werk Ruralia commoda von Petrus de Crescentiis. An der ULB Tirol befinden sich mehrere Wiegendrucke von Augustinus’ „De civitate Dei“: GW 2879 (Venedig: Nicolas Jenson, 2. Oktober 1475), GW 2880 (Venedig: Gabriele di Petro, 1475), GW 2883 (Strassburg: Johann Mentelin, nicht nach 1468), GW 2887 (Basel: Johann Amerbach, 13. Februar 1489). Auch die Werke des Flavius Iosephus (GW M15160 – Augsburg: Johann Schüssler, 28. Juni und 23. August 1470) sowie ein Exemplar der „Ruralia commoda“ (GW 7820 – Augsburg: Johann Schüssler, 16. Februar 1471) werden an der ULB Tirol aufbewahrt. Daneben finden sich auch eine Sermonessammlung, wohl die „Sermones aurei de sanctis“, von Leonardus de Utino (GW M17891 – Nürnberg: Anton Koberger, 22. Jänner 1478; an der ULB Tirol vorhanden) und die weit verbreitete „Summa de casibus conscientiae“ des Bartholomaeus de Sancto Concordio. Bei Letzterer ist sogar eine Jahreszahl beigefügt: 74. Es ist also anzunehmen, dass es sich um die Ausgabe, die am 17. Jänner 1474 von Arnold ter Hoernen in Köln gedruckt wurde (GW 3451), handelt. Ein Exemplar dieser Ausgabe ist weder in Neustift noch an der ULB Tirol erhalten; in Innsbruck wird ein Exemplar GW 3453 (Augsburg: Günther Zainer, 1475) und eines GW 3455 (Speyer: Peter Drach d. Ä., nicht nach 1479) aufbewahrt. In dasselbe Jahr datiert auch ein nicht näher definierter kleiner, aus dem Griechischen ins Lateinische übersetzter Traktat, der gemeinsam mit einem großformatigen Buch mit dem Titel Planctus ecclesie um 35 Gulden erworben wurde. Letzteres Werk ist möglicherweise zu identifizieren als „Planctus ruinae ecclesiae“ von Iohannes Fabri; allerdings sind laut GW nur Ausgaben in Quartformat erhalten; weder für Neustift noch für die ULB Tirol ist eine entsprechende Ausgabe nachgewiesen.Weitere 20 Gulden waren vonnöten, ad eundem illigandum et corpora faciendum, also um das Buch binden zu lassen. Breviarium Salisburgense. Venedig: Nikolaus von Frankfurt, 1482 (GW 5442). Vgl. u. a. M. Barczyk, P. Schurer, Kirche und Stift St. Peter zu Waldsee (Veröffentlichungen des Stadtarchivs Bad Waldsee B/3). Bad Waldsee 1979 sowie Eintrag „Augustiner Chorherrenstift Waldsee“ in der Datenbank „Klöster in Baden-Württemberg“ (https://www.kloester-bw.de/index.php). Vgl. P. A. Beck, Lukas Härber, Probst von Waldsee und Neustift: ein Beitrag zur Geschichte des Chorherrenstifts Waldsee. Diözesan-Archiv von Schwaben 18 (1900) 97–101. Vgl. Katalog ULB Tirol X (wie Anm. 100) 469–472. A. Giner, Chorherrenverzeichnis 1142–1942, in: A. Giner (unter Mitwirkung von A. Sparber, M. Schrott) (Hrsg.), Festschrift zum 800jährigen Jubiläum des Stiftes Novacella. Brixen 1942, Anhang 1–61, hier 12. 18 Die Stiftsbibliothek von Neustift Wahrscheinlichkeit stammen auch ein Evangelistar und Lektionar (Cod. 190) aus seiner Hand. Die Frage, ob Stetner auch als Buchmaler tätig war, wie wohl aufgrund der Signatur in Cod. 194 auf Bl. 70v, das eine ganzseitige Miniatur zeigt, in der Literatur diskutiert wurde36, kann mit ziemlicher Sicherheit verneint werden. Die kulturelle Blütezeit in Neustift erlebte am 12. Mai 1525 ein jähes Ende: Aufständische Bauern aus der Umgebung, angeführt von Michael Gaismair, plünderten das Stift und richteten, wie der Chro nist und Hofrichter von Neustift Georg Kirchmair von Ragen berichtet, großen Schaden an Prief vnd Puechern37 an. Den Bauern gelang es zwar nicht, die gesuchten Urbare mit den darin festgehaltenen Abgabenverpflichtungen an sich zu reißen, doch hinterließen sie eine Spur der Verwüstung: … in summa alle pücher was [?] geschriben gewest ist aus allen cästn, truchn, stubn sonderlich canzlei stübl schuldbrief alles zerissen, auf den hof tragen, zerhackt, zerschnidten, zerissen, zerstochn auf die spieß und hellpardt gelegt und gegeneinannder geworfen …38 Welche Dokumente und Bücher ihnen tat sächlich in die Hände fielen und zerstört wurden, entzieht sich unserer Kenntnis. Überliefert wird, dass das sogenannte Posch-Missale (Cod. 100) nur deswegen nicht zerstört worden sei, da es Stetner als Pfarrer von Natz im dortigen Widum und nicht im Stift aufbewahrt hatte. Als Beweis hierfür wird gemeinhin die Datierung auf der ganzseitigen Miniatur auf Bl. Iv angeführt: 1526. Das Messbuch habe demnach ein Jahr lang im Widum von Natz gelegen, ehe es – wohl von einem nicht näher bekannten Wanderminiator – detailreich und aufwändig illuminiert wurde. Der Aufstand konnte zwar nach drei Tagen niedergeschlagen werden, aber das Stift erholte sich nur langsam von diesem Einfall und den Plünderungen. In der zweiten Hälfte des 16. Jahrhunderts ließen die Pröpste Augustin II. Schabl (1569–1581) und seine Nachfolger Augustin III. Distelfink (1589–1589) sowie Jakob Fischer (1589–1621) zahlreiche Handschriften und Bücher neu binden. Ob dies in Zusammenhang mit den Verwüstungen während des Bauernaufstandes zu sehen oder einer Neuaufstellung geschuldet ist, bleibt offen. Unter den von Propst Augustin Distelbrink getätigten Aus gaben finden sich auch mehrere Buchkäufe bei einem Buchführer namens Albrecht in Brixen39 sowie Entgelte für überreichte Widmungsschriften.40 Besondere Erwähnung verdient ein Eintrag vom 10. März 1588, zumal er anschaulich belegt, dass auch gegen Ende des 16. Jahrhunderts besondere Werke weiterhin selbst hergestellt wurden: Am 10. dem Steffan Wenzl khauffmann zu Braunegg der mir das pergamen, auch rot unnd plabe farben zu unser mespuech hergeben hat ime völlig bezalt 33 fl. 56 kr.41 Ein Messbuch auf Pergament aus jener Zeit ist nicht auf uns gekommen – auch dies wohl einer seits ein Beispiel, wie bruchstückhaft unsere Kenntnisse über die Neustifter Schreibstube und deren Erzeugnisse sind, andererseits aber auch ein Beleg dafür, dass das handgeschriebene Buch auch nach Erfindung des Buchdrucks keineswegs seine Berechtigung verloren hatte.42 Gerade für den gemein 36 37 38 39 40 41 42 Vgl. hierzu das Katalogisat zu Cod. 194 im vorliegenden Band. Zit. nach Th. G. v. Karajan (Hrsg.), Johannes Tichtels Tagebuch, Sigmunds von Herberstein Selbstbiographie, Johannes Cuspinians Tagebuch, Georg Kirchmairs Denkwürdigkeiten (Fontes Rerum Austriacarum / Österreichische Geschichts-Quellen I, 1). Graz 1855, 472. Zit. nach K. Pattis, Neustift zur Zeit des Bauernaufstandes. Wirtschaftliche, soziale und religiöse Hintergründe. Brixen 2012, 35. Neustift, Stiftsarchiv, Cod. 26, S. 22 (2. Juli 1585): Von Albrecht Puechfüerer zu Brichsen die thayll jurii khauff 5 fl.; S. 76 (8. Jänner 1587): Am 8 dits dem Albrecht buechfüerer zu Brixen fur etlich büecher bezalth 3 fl. 12 kr. Im September desselben Jahres folgte noch der Kauf einer Postille, eines Gebetbuches und einer lanornung (?), vgl. ebda, S. 86, 88. Neustift, Stiftsarchiv, Cod. 26, S. 3 (6. März 1585): Den herrn Donat alda zu Brichsen, der zu meiner consecration ministriern helffen, auch etlicher carmina halber die er mir dediciert und druckht hat verehrt 5 fl.; S. 33 (17. September 1588): Ainem studenten der mir carmine dediciert geben 30 kr. Neustift, Stiftsarchiv, Cod. 26, S. 101. Sehr deutlich wird auf die weiterhin große Bedeutung des handschriftlichen Buches auch in einer Neustifter Bibliotheksordnung, überliefert als Abschrift im frühen 18. Jahrhundert durch den Chorherrn Petrus Ephensteiner, hingewiesen: „Trotz der Erfindung des Buchdrucks soll man im Kloster das Bücherschreiben als eine echt monastische Aufgabe beibehalten und dabei mehr auf Lesbarkeit und Dauerhaftigkeit der Schriften als auf äußeren Glanz bedacht sein.“ Die Bibliotheksordnung befindet sich im Stiftsarchiv, allerdings ohne Die Stiftsbibliothek von Neustift 19 schaftlichen Chorgesang erfüllte möglicherweise aufgrund des notwendigen Formats in Übergröße b isweilen ein Codex besser die Bedürfnisse als ein Druck, dessen Format aus technischen Gründen limitiert war. Zudem galt das Bücherschreiben auch weiterhin als besondere Form des „Gottes Diens tes“. Insbesondere bei liturgischen Büchern machten auch nach der Einführung des einheitlichen Römischen Ritus lokale Besonderheiten eine manuelle Anfertigung erforderlich. So datiert das mit einer Höhe von 82 cm und einer Breite von 55 cm größte erhalten gebliebene Choralbuch in die 1620er Jahre.43 Laut Chorherrenverzeichnis hat auch Michael Schwarzkröll (1628–1674) „einige Choralbücher geschr[ieben]“44, vom Chorherrn Gregor Oberpla(t)zer (1643–1701) ist ein zweibändiges Werk in Großfolio-Format mit aufwändig gestalteten Initialen erhalten.45 Die seit der zweiten Hälfte des 16. Jahrhunderts zunehmende wissenschaftliche Betätigung der Chorherren46 sowie die Errichtung einer stiftseigenen theologisch-akademischen Hauslehranstalt mit dem Ziel der Verbesserung der Ausbildung von Novizen und Klerikern als Folge des Trienter Konzils trugen zweifelsohne zu einem großen Bücherzuwachs bei. „Bereits seit dem 16. Jahrhundert war hier ein umfangreicher Bestand an theologischen Büchern und Lehrwerken für die Bibliothek angeschafft worden, zum Teil das Beste, das aus den Druckereien in Nürnberg, Basel, Rom und Lyon zu haben war.“47 Besondere Verdienste um die Hauslehranstalt erwarb sich Propst Markus Hauser (1621–1665). Er holte zahlreiche gelehrte Ordensmänner aus anderen Klöstern nach Neustift, darunter auch den Jesuiten Adam Weber. Innerhalb kurzer Zeit hatte Neustift als Studienort einen so guten Ruf, dass viele Kleriker aus anderen Klöstern dorthin kamen. Von dieser Zeit und dem hohen Niveau der Hauslehr anstalt, die bis 1807 Bestand haben sollte, legen die erhalten gebliebenen Thesenblätter, die anlässlich des Examens und der Defensio hergestellt und verteilt wurden, beredtes Zeugnis ab.48 Zahlreiche Werke, darunter der 1517 in Nürnberg bei Johann Schönsperger d. Ä. gedruckte und reich illustrierte „Theuerdank“ (Barockbibliothek, Sign. 5)49, gelangten als Geschenke in die Neustifter Bibliothek, zudem wurden Bibliotheken von Verstorbenen den Stiftssammlungen einverleibt. Sämtliche Bücher des Grafen Ulrich von Enzenberg wurden z. B. nach seinem Ableben aufgrund der verwandtschaft lichen Beziehungen zu Propst Fortunat Troyer (1678–1707) in die Neustifter Bibliothek eingereiht.50 Seit dem 17. Jahrhundert, als aufgrund des großen Bücherzuwachses und insbesondere der v ermehrten Benutzung der Bibliothek eine verstärkte Sorge um die Buchpflege, die Ordnung und Auf stellung sowie um eine systematische Erwerbung vonnöten war, sind auch Namen der Stiftsbibliothekare von Neustift bekannt. Es ist zwar davon auszugehen, dass das Offizium seit jeher besetzt war – schließ lich fand der Umgang mit Büchern sogar Eingang in die Ordensregel des hl. Augustinus (V, 9–10) –, 43 44 45 46 47 48 49 50 Hinweis auf Petrus Ephensteiner. Zur Zuordnung samt Abschrift vgl. M. Peintner, Die geistesgeschichtliche Bedeutung der Stiftsbibliothek, in: 850 Jahre Augustiner Chorherrenstift Neustift (wie Anm. 4) 127. M. Peintner, Schreibkunst, Studium und Musikleben im mittelalterlichen Kloster. Eine Darstellung am Bei spiel der Musikgeschichte des Augustiner-Chorherrenstifts Neustift, in: K. Drexel, M. Fink (Hrsg.), Mu sikgeschichte Tirols I: Von den Anfängen bis zur Frühen Neuzeit (Schlern-Schriften 315). Innsbruck 2001, 353–380, hier 372. Chorherrenverzeichnis (wie Anm. 35) 20. Peintner, Die geistesgeschichtliche Bedeutung der Stiftsbibliothek (wie Anm. 42) 112–133, hier 128. Während für das 15. und 16. Jahrhundert nur vereinzelt Nachrichten von Neustifter Chorherren an Universitäten erhalten sind, lassen sich seit 1600 mehrere Neustifter Kleriker an den Universitäten Dillingen, Ingolstadt und Salzburg nachweisen; vgl. Th. (H). Innerhofer, 850 Jahre Erziehung und Unterricht, in: 850 Jahre Augustiner Chorherrenstift Neustift (wie Anm. 4) 154–167, hier 160. F. Brendle, Die Augustiner-Chorherren, in: F. Jürgensmeier, R. E. Schwerdtfeger (Hrsg.), Orden und Klöster im Zeitalter von Reformation und Katholischer Reform 1500–1700, Bd. 3 (Katholisches Leben und Kirchenreform im Zeitalter der Glaubensspaltung 67). Münster 2007, 39–64, hier 60. Innerhofer, 850 Jahre Erziehung (wie Anm. 46) 162. Der „Theuerdank“ soll dem Stift von Erzherzog Maximilian III. von Österreich als Dank für die Aufnahme des Hofes in Neustift während der Fleckfieberepidemie in Innsbruck im Herbst 1611 überreicht worden sein. C. Schretter-Picker, L. Šubarić, U. Stampfer, W. Neuhauser, Barocke Buchkultur im Tiroler Raum, in: E. Klecker, Ch. Gastgeber (Hrsg.), Buchkultur des Barock (Geschichte der Buchkultur VII). Graz 2015, 295–382, hier 347. 20 Die Stiftsbibliothek von Neustift doch war das Amt des bibliothecarius vorab meist mit anderen Aufgaben verbunden51 und erlangte erst durch die deutliche Vergrößerung der Bestände an Bedeutung, sodass dies auch vermehrt in Quellen vermerkt wurde. So hatten Martin Warell (1665), der insbesondere als Stiftsarchivar und durch die von ihm veranlasste, bis heute gültige Ordnung des Archivbestandes Bekanntheit erlangte52, Christoph Ach müller (1679), Anton Steiger (1687–1693), Markus von Sayburg (1706), Matthias Alban (1709), Hieronymus Kon (1735), Dominikus Pussieger (1735), Bartholomaeus Perathoner (1735), Martin Raith (1741), Caspar Mayr (1742), Philipp Neri Puell (1758), Johannes Chrysostomus Pertinger (1764, 1769), Alois von Lidl (1767), Stanislaus Pertinger (1782), Franz Xaver Grass (1787, 1828), Lorenz Hueber (1798), Ferdinand Steiner (1799) und Gaudenz Amort (1800) dieses Offizium inne.53 Der enorme Bücherzuwachs machte eine Vergrößerung der Bibliotheksräumlichkeiten bzw. eine Neuorganisation der Sammlung bald unumgänglich. Der Wunsch nach Repräsentation und der Anspruch, barockes Selbstbewusstsein zur Schau zu stellen, erforderten einen Neubau an prominenter Stelle. Propst Leopold de Zanna (1767–1787) ließ im Süden des Stiftshofes anstelle des domus capellanorum54 einen dreigeschossigen Bau aufführen, der der reichen Büchersammlung des Stiftes angemessene Räumlichkeiten bot und den Stiftshof nach Süden hin abschloss. Der Baumeister Antonio Giuseppe Sartori aus Sacco bei Rovereto, der als einer der besten Tiroler Architekten seiner Zeit galt, folgte dem Beispiel zahlreicher anderer Stifte und errichtete in den Jahren 1771 bis 1778 einen doppel stöckigen Saal mit umlaufender Galerie, die man durch zwei wie kleine Türmchen in den Ecken gestaltete Wendeltreppen erreicht – in den anderen beiden befinden sich Drehschränke. Der 11 × 23 m große Saal wurde von Hans Mussack aus Sistrans bei Innsbruck mit reichen Stuckaturen in Weiß und Gold versehen, die ihm einen äußerst festlichen Charakter verleihen. Dazu tragen auch die aufwändigen Portale mit Intarsien und der Natursteinboden in Weiß, Schwarz und Rot mit Mittelrosette wesentlich bei. Die 42 Bücherschränke, jeweils mit einem geschnitzten Aufsatz für die Titel der unterschiedlichen Sachbereiche versehen, sind an die Wände zwischen den Fenstern gestellt, im Raum selbst befanden sich lediglich zwei lange Schranktische, die Platz für besonders große Bücher und möglicherweise wissenschaftliches Instrumentarium wie Globen boten. Der Saal konnte so für Veranstaltungen wie öffentliche Disputationes und repräsentative Zwecke genutzt werden. Insgesamt fanden im Saal, der auf jeder Längsseite von zehn Fenstern großzügig belichtet wird, ca. 20.000 Bände Platz. Der räum lichen Gliederung entsprechend waren die verschiedenen theologischen Disziplinen im Untergeschoss und die Werke aus dem Bereich der Humanwissenschaften auf der Galerie deutlich voneinander getrennt. Die Aufstellung nach Größen und die weitestgehende Vereinheitlichung der Einbände – viel fach wurden die alten Einbände auch lediglich mit einem neuen Rücken aus braunem Kalbsleder mit einer Titelaufschrift in Gold versehen – sorgten für einen geschlossenen Eindruck und betonten den Repräsentationsanspruch. 51 52 53 54 So ist der als Archivar und wohl auch Bibliothekar tätige Chorherr Johannes Librarius (gest. 1467), der um 1465 das „Memoriale benefactorum“ verfasst hat, auch als Johannes Cellerarius nachgewiesen, vgl. A. Dörrer, Johannes Librarius, in: Die deutsche Literatur des Mittelalters. Verfasserlexikon, begründet von W. Stammler, hrsg. von K. Ruh (u. a.), IV. Berlin, New York 1983, Sp. 672–673; J. Riedmann, F. Schaffenrath, Geschichtsschreibung, in: M. Korenjak, F. Schaffenrath, L. Šubarić, K. Töchterle (Hrsg.), Tyrolis Latina. Geschichte der lateinischen Literatur in Tirol I: Von den Anfängen bis zur Gründung der Universität Innsbruck. Wien, Köln, Weimar 2012, 105–122, hier 115–117. L. Šubarić, F. Schaffenrath, P. Kennel, Geschichtsschreibung, in: M. Korenjak, F. Schaffenrath, L. Šubarić, K. Töchterle (Hrsg.), Tyrolis Latina. Geschichte der lateinischen Literatur in Tirol II: Von der Gründung der Universität Innsbruck bis heute. Wien, Köln, Weimar 2012, 726–777, hier 727–728. Vgl. Chorherrenverzeichnis (wie Anm. 35) 13–37. Nachdem Propst Hieronymus Rottenpuecher 1667 das damalige Schulhaus im Mittelhof hatte abreißen lassen, waren in diesem Kaplanhaus das Schulzimmer und die Lehrerwohnung untergebracht; vgl. Innerhofer u. a., Von der Klosterschule (wie Anm. 5) 15–17; A. Sparber, Aus der inneren Geschichte unseres Klosters, in: Festschrift zum 800jährigen Jubiläum (wie Anm. 35) 69–139, hier 115; B. Rutz, Die Chorknaben zu Neustift. Ein Beitrag zur Geschichte der Schule und der Musik in Tirol. Innsbruck 1911 (Separatdruck aus Neue Tiroler Stimmen), 22. Die Stiftsbibliothek von Neustift 21 Kurze Zeit nach Fertigstellung dieses Saales besuchte Adalbert Blumenschein55, der in der zweiten Hälfte des 18. Jahrhunderts Bibliotheksreisen durch ganz Europa unternahm, Kloster Neustift. Seinen Reisebeschreibungen ist zu entnehmen: „Dieses von justerwehnter Stadt [Brixen] nur eine Stunde ent legene dennen Canonic. Regularib. S. Augustini angehörige Stüfte besizet eine sehr gute Bibliotheck. Das Behältniß der Bücher ist ganz neu erbauet. Sie steht in einem viereckigtem zimlich breiten, jedoch von einer nicht sonderlichen Höhe aufgeführten Saale, welcher von beiden Seiten unter dem an der Wand herumlaufend holzernen Gange durch zehn grosse, und oberhalb durch eben so viele kleinere Fenster ungemein hell erleuchtet wird. Die flache Oberdecke ist zwar nicht gemalen, jedoch mit sehr schön vergoldeter Stukatorarbeit nach der neuesten Art gezieret. Die Repositorien sind weis ange strichen, und mit Firniß überzohen, sodann die Leisten, und Ausschnitte ebenfals gut übergoldet, und weilen auch überhin die Bücher meist französisch gebunden, so läßt Solches vortreflich. Dieses Behält niß hat nun seinen Ein= und Ausgang sowohl von der Probstey, als von dem Convent.“56 Als Arbeitsund Studierräume waren unmittelbar an den großzügigen Saal angeschlossene Räume und Kabinette vorgesehen, die zudem noch reichlich Platz für weitere Bücher boten. „Belangend nun den hiesig ganzen Bücherschaz zusammen, so schäz ich Sölchen ungefähr etwas über 8000: Bände, dann 20000: wie mir Sölcher von dennen hiesigen Geistlichen angegeben worden, ist in der Thatt zu viel.“57 Die Einrichtung von kleineren Nebenräumen entsprach auch der Forderung, die bereits in einer älteren Neustifter Bibliotheksordnung, erhalten als Abschrift des Chorherrn Petrus Ephensteiner (1691–1739), nachzulesen ist: „Neben der Hauptbibliothek soll auch eine Handbibliothek da sein, damit jene Werke, die häufig gebraucht werden, auch stets zur Verfügung stehen.“58 In diesem Schreiben werden auch die Aufgaben eines Bibliothekars genau festgehalten: „Der Bibliothekar hat die Bücher vor allem vor Feuchtigkeit und Regen zu schützen. An schönen Tagen soll die Bibliothek fleißig gelüftet werden. Die Bücher sollen nach ihrem Inhalt in verschiedenen Schränken einen bestimmten Standort haben. Der Bibliothekar soll ein Verzeichnis haben, in dem die vorhandenen Werke eingetragen sind, und er soll dem Prälaten Rechenschaft geben, so derselbe diese verlangt. Jedes einzelne Buch soll seinen Titel tragen und am Ende einen Index. Desgleichen sind die Erwerbsdaten einzutragen und der Vermerk: Bibliotheca Collegii B.M.V. ad gratias Neocellae.“59 In der Tat wurde der Großteil der Druckwerke und Handschriften in Neustift mit einem Bucheignerzeichen, in Holzschnitt technik gefertigt, ausgestattet; am häufigsten vertreten ist ein Exlibris mit Zierrahmen und dem Ver merk Bibliothecae Novacellensis samt einem T (Tau-Kreuz), dem Wappenzeichen des Stiftes.60 In den Handschriften sind meist auch noch ältere, handschriftliche Eintragungen zu finden: Der Großteil der Besitzvermerke stammt aus dem 15. Jahrhundert und weist stets einen ähnlichen Wortlaut auf.61 Als zentrale Aufgabe des Bibliothekars wird des Weiteren die Erstellung von Bibliotheksverzeich nissen und Katalogen zur besseren Orientierung und zur Übersicht über die Bestände genannt. Aus früheren Zeiten sind keine Verzeichnisse oder Listen überliefert, erst der Neustifter Bibliothekar und 55 56 57 58 59 60 61 Vgl. G. Osti, Adalbert Blumenschein: l’uomo e l’opera. Atti della Accademia Roveretana degli Agiati 250 (2000), ser. VII, vol. X, A, 269–337. Zit. nach W. Neuhauser, Blumenscheins Bibliotheksreisen durch Tirol (Nordtirol und Südtirol). Atti della Accademia Roveretana degli Agiati 250 (2000), ser. VII, vol. X, A, 339–415, hier 347. Ebda. Anschließend folgt eine kurze Beschreibung der vorhandenen Handschriften (Blumenschein hält allerdings gleich zu Beginn fest: „Von alten MSS: ist zwar alda nichts Sonderliches vorfindig“) sowie der Inkunabeln („Von dennen gedruckten Wercken aber aus dem XV: Jahrh: sind mehr dann 300: Stück zugegen“). Vgl. Anm. 42. Vgl. ebda. Zum Stiftswappen vgl. F.-H. v. Hye, Wappen in Tirol: Zeugen der Geschichte. Handbuch der Tiroler Heraldik (Schlern-Schriften 321). Innsbruck 2004, 169–171; H. G. Ströhl, Die Wappen der Ordensstifte in Tirol und Vorarlberg. Kunst und Kunsthandwerk XV/4 (1912) 193–196. Vgl. Anm. 120 und 122. 22 Die Stiftsbibliothek von Neustift Orientalist Franz Xaver Grass (1758–1833) verfasste Ende des 18. Jahrhunderts gleich mehrere der artige Verzeichnisse. Es waren dies die ersten gedruckten Bibliotheksverzeichnisse im Tiroler Raum.62 Nur wenige Jahre nach Fertigstellung der neuen Bibliothek, die der reichen Neustifter Bücher sammlung einen angemessenen Raum bot, folgte eine schwere Zeit, die dem Stift stark zusetzte. Kloster Neustift blieb zwar von den Josephinischen Klosteraufhebungen verschont, doch wurde es während der drei Koalitionskriege gegen Frankreich (1792–1805) zwischenzeitlich als Lazarett benutzt und hatte immer wieder umfangreiche Einquartierungen von Truppen beider Kriegsparteien zu erdulden63 und drückende Abgaben zu leisten. Nachdem Tirol schließlich 1806 den Bayern zugesprochen worden war, folgte am 17. September 1807 die Aufhebung aller noch bestehenden tirolischen Klöster und Stifte. Sämtliche Kostbarkeiten, Kunstgegenstände und Bücher mussten abgeliefert werden, die Stiftsgüter wurden veräußert bzw. verpachtet. Bereits im November desselben Jahres wurden die ersten wertvollen Paramente, das Haus- und Kirchensilber sowie andere Pretiosen von Neustift nach Innsbruck ge schickt.64 Auch das gotische Chorpult wurde aus Neustift entfernt.65 Für die Bücher sollte hingegen, ebenso wie für die Tafelbilder und Gemälde, vor Ort eine Auswahl getroffen werden. Der kurfürstliche „Bilder-Galerie-Inspector“ Johann Georg von Dillis reiste daher im Sommer 1808 aus München an, um in Neustift und anderen Klöstern besonders wertvolle Bilder für den Abtransport auszusuchen (u. a. den Kirchenväter-Altar von Michael Pacher); bei der Auswahl der Druckwerke und Handschriften sollte Franz Xaver Grass behilflich sein. Als dieser sich jedoch weigerte – „offenbar blutete ihm darob das Herz“66 –, wurde hiermit der Chorherr Hermann Mader beauftragt. Wegen der umfassenden, kurz vorher von Grass erstellten Kataloge konnten aber keine Raritäten und Kostbarkeiten der Bibliothek übergangen werden. Ein genaues Verzeichnis über die ausgewählten Bücher wurde allerdings im Gegensatz zu anderen Klöstern67 nicht erstellt. Nicht weniger als 60 alte Zentner (ca. 3 Tonnen) Bücher, darunter zahlreiche Handschriften und Inkunabeln, bzw. 20 Kisten wurden schließlich am 7. und 8. April 1809 an die Universitätsbibliothek nach Innsbruck überstellt. Ob einige Bücher auch in die Hofbibliothek nach München oder anderswohin gelangten, lässt sich aufgrund des Fehlens einer genauen Liste nicht mit Sicherheit feststellen. Eine entsprechende Überprüfung vor Ort ist noch aus ständig. Zudem wurden die reiche Mineralien- und Elfenbeinsammlung, das Münzkabinett und allerlei sonstige Besonderheiten, die sich – einer Kunst- und Wunderkammer gleich – im Vorraum der Biblio thek befunden hatten, aus dem Stift entfernt. 62 63 64 65 66 67 F. X. Grass, Raritas Librorum In Bibliotheca Novacellensi Canonicorum Regularium S. Augustini Delitescentium Luci Publicae Exposita … Brixinae 1777; Ders., Verzeichniß typographischer Denkmäler aus dem fünfzehenten Jahrhundert, welche sich in der Bibliothek des regulirten Korherrnstiftes des heil. Augustin zu Neustift in Tyrol befinden. Brixen 1789; Ders., Verzeichniß einiger Büchermerkwürdigkeiten aus den sechzehenten und siebenzehenten Jahrhunderten, welche sich in der Bibliothek des regulirten Korherrenstiftes des heil. Augustin zu Neustift in Tyrol befinden. Brixen 1790; Ders., Nachtrag zu den typographischen Denkmälern aus dem fünfzehenden und Büchermerkwürdigkeiten aus dem sechzehenden und siebenzehenden Jahrhunderte, welche sich in der Bibliothek des regulirten Korherrenstiftes des heil. Augustin zu Neustift in Tyrol befinden. Brixen 1791. A. Sparber, Aufhebung und Wiederherstellung des Stiftes (1807–1816), in: J. Huber (Hrsg.), Aus der Chronik des Chorherrenstiftes Neustift bei Brixen. Neustift 1956, 66–105, hier 67. Ebda 73–78. Ein Foto des Chorpultes findet sich in: Festschrift zum 800jährigen Jubiläum (wie Anm. 35) Bild 47. Es ist in privatem Besitz und befindet sich heute als Dauerleihgabe im Bayerischen Nationalmuseum in München. Sparber, Aus der inneren Geschichte (wie Anm. 54) 115. Laut Sparber konnte sich Grass „wegen Arbeitsüberbürdung, nämlich als Pfarrer von Natz“ entschuldigen (Sparber, Aufhebung [wie Anm. 63] 89). Als Beispiel sei hier der an der ULB Tirol aufbewahrte 80 Blatt starke Übergabekatalog der Kartause Schnals aus dem Jahr 1784 genannt (Innsbruck, ULB Tirol, Cod. 915). Die Stiftsbibliothek von Neustift 23 Als das Stift am 12. Jänner 1816 mit kaiserlichem Edikt wiedererrichtet wurde68, hat man wenig später auch die Rückführung der Bücher angeordnet.69 Gerade weil an die Wiedererrichtung u. a. die Bedingung geknüpft war, dass die Neustifter Chorherren Lehrtätigkeit auszuüben bzw. sämtliche Lehr kräfte für das k. k. Gymnasium in Brixen zu stellen hatten70, war man um eine möglichst rasche Rück führung bemüht. Sie gestaltete sich jedoch als äußerst schwierig. Obwohl Propst Leopold Erlacher (1790–1832) wiederholt […] um Zurückgabe der seinem Stifte angehörigen bey der Bibliothek allhier befindlichen Bücher71 gebeten und sogar bei der „Landes Stelle“ vorgesprochen hatte, tat der damalige Bibliothekar an der Universitätsbibliothek, Johann Bertholdi72, alles, um die Rückstellung hinauszu zögern bzw. um nicht den gesamten Bestand zurückgeben zu müssen. Er soll erklärt haben, der Kaiser könne nicht mehr zurückgeben, was der König von Bayern einmal der Universitätsbibliothek geschenkt habe.73 Schließlich bot Propst Erlacher an, die bereits der k. k. Lycealbibliothek eingereihten Bücher dortzubelassen – wohl in der Hoffnung, so die anderen schneller zu erhalten; doch selbst die Anwei sung der Tiroler Landesregierung an Bertholdi, dem Bevollmächtigten des Herrn Prälaten zu Neustift ungesäumt alle Werke des Stiftes Neustift zu übergeben, welche noch nicht den Katalogen eingereiht worden und zur klösterlichen Lektüre geeignet sind 74, blieb ohne Wirkung. Wie ein Schreiben des Landeshauptmanns von Tirol, Ferdinand von Bissingen-Nippenburg, an Bertholdi vom Mai 1818 deutlich macht, erklärte sich Propst Erlacher vielmehr zu einem neuerlichen Geschenk bereit und ver zichtete auf die typographischen Denkmäler aus seiner Bibliothek; alle anderen Bücher sollten aber sobald möglich75 ausgehändigt werden. Dass die Rückstellung immer noch nicht erfolgt sei, habe laut Bertholdi aber daran gelegen, dass der Propst die Bücher nicht habe abholen lassen.76 Es sollten schließ lich noch mehr als zehn Jahre vergehen, ehe die Bücher dann auch tatsächlich im August 1833 auf Betreiben des Propstes Ludwig Mair (1832–1851) und vor allem nach Einschreiten des Guberniums77 68 69 70 71 72 73 74 75 76 77 Vgl. Wien, Haus-, Hof- und Staatsarchiv, Kabinettsarchiv, Staatsrat, Protokolle und Indizes, 241 Protokoll 1815: […] Ich will die vier Stifter der Prämonstratenser zu Wilten, der Zisterzienser in Stams, der regulirten Chorherrn zu Neustift u. Benediktiner in Marienberg beibehalten, und genehmige die in dieser Hinsicht gemachten Anträge der Z: O: Hofkommission mit dem Beisatze, daß diese Stifter stets bemüht seyn sollen, einen an Talenten, Fleiß und Sitten sich auszeichnenden, wohlgebildeten Nachwuchs zu bekommen, um ihn außer der Seelsorge, vorzugsweise auch bei den öffentlichen Lehr- und Erziehungsanstalten mit Beruhigung verwenden zu können […], dat. Mailand, 12. Jänner 1816 (freundlicher Hinweis David Fliri, Wien). Vgl. Innsbruck, ULB Tirol, Direktionsakt 1816, Nr. 204, 22. Februar 1816. Sparber, Aufhebung (wie Anm. 63) 95. Innsbruck, ULB Tirol, Direktionsakt 1817, Nr. 209, 13. Februar 1817. Johann Bertholdi war 1806 als Bibliothekar von der bayerischen Regierung eingesetzt und 1814 von der österreichischen in seinem Amt bestätigt worden (vgl. W. Neuhauser, Die Geschichte der Handschriftensammlung der UB Innsbruck, in: Ders. [Hrsg.], Handschriftenkunde [wie Anm. 23] 51–72, 64; A. Hittmair, Geschichte der k. k. Universitätsbibliothek in Innsbruck, Innsbruck 1910 [Sonderabdruck aus: Zeitschrift des Ferdinandeums für Tirol und Vorarlberg 3/54], 73–74). Zit. nach Hittmair, Geschichte (wie Anm. 72) 85. Innsbruck, ULB Tirol, Direktionsakt 1817, Nr. 218, 4. Dezember 1817. Innsbruck, ULB Tirol, Direktionsakt 1818, Nr. 220, 14. Mai 1818. Innsbruck, ULB Tirol, Direktionsakt 1819, Nr. 227, 30. Juni 1819 (Antwortschreiben auf eine Aufforderung von Seiten der Landesverwaltung vom 27. Mai 1819, der Landesstelle [wohl Studienhofkommission] ein Verzeichnis der an die Stifte zurückgegebenen Bücher vorzulegen). Die Landesstelle kann keinen Anstand nehmen, dem Stifte Neustift jene Bücher zurückstellen zu lassen, welche sich von diesem Stifte in der hiesigen Universitätsbibliothek befinden und dermalen fortan von eben diesem Stifte noch angesprochen werden. Der Umstand, daß diese Werke bereits in die Kataloge eingereiht, und daß sie der Universitätsbibliothek wichtig sind, kann ebenso wenig, als die seit dem Jahre 1817 verzögerte Hinausgabe dieser Bücher als entscheidend, oder hemmend angesehen werden, ob aber die inzwischen eingetretenen Verfügungen mit den Duplikaten an der hiesigen Universitätsbibliothek nicht etwa eine, und welche Schwierigkeit in Ausführung dieser Rückgabe begründen, wird sich durch die weitere Verhandlung zeigen. Der Herr Universitätsbibliothekär erhält daher den Auftrag, alsobald, als es die Jahreszeit erlaubt, anzuzeigen, ob sich die im schriftlichen Verzeichniße, welches vom Stifte Neustift bei Gelegenheit der Inventur am 16. Juli v. J. vorgelegt, und welches dem Herrn Universitätsbibliothekär mit Gubernialdekret vom 4ten v. M. Zahl 28167 übergeben wurde, aufgeführten Bücher noch alle vorfinden, und wohin die etwa abgängigen gekommen seyen. 24 Die Stiftsbibliothek von Neustift wieder nach Neustift gebracht wurden – nicht ohne vorher noch mehrmals um die Überlassung einiger Bücher zu bitten.78 Bitten, denen das Stift auch nachkam.79 Als Vincenzo Tizzani, General-Prokurator des Augustiner Chorherrenordens, 1842 anlässlich des 700-Jahr-Jubiläums nach Neustift reiste und die dortige Bibliothek besichtigte, fand er dort wieder eine stattliche Zahl an Büchern vor: „Sie ist reich an Büchern, welche nach dem Inhalte geordnet sind: die religiösen Inhaltes bilden bei Weitem den wichtigsten Theil. […] Der Bibliothek fehlt der Catalog, der aber nächstens wird verfertiget werden. Die Zahl der Bücher möchte die Nummer 9000 erreichen.“80 Während die „typographischen Denkmäler des 15. Jahrhunderts“, d. h. die Inkunabeln, bereits 1819 der Universitätsbibliothek als Geschenk überlassen wurden – was auch im Schriftwechsel 1833 immer wieder Erwähnung fand –, werden die mittelalterlichen Handschriften nie eigens genannt. Erst als nach dem Ersten Weltkrieg Südtirol Italien zugesprochen wurde, gewann die Frage nach der Rück gabe von Kulturgütern erneut an Bedeutung. Der Friedensvertrag von Saint-Germain-en-Laye vom 10. September 1919 sah klare Bestimmungen hierfür vor.81 Die Republik Österreich verpflichtete sich, sämtliche Kunstgegenstände und expressis verbis auch Handschriften, die in Verletzung des Rechtes 78 79 80 81 Damit erlediget man vorläufig den Bericht des k. k. Herrn Universitätsbibliothekärs vom 19. v. Mts Zahl 47. Innsbruck am 11. Jänner 1833 (Innsbruck, ULB Tirol, Direktionsakt 1833, Nr. 345–67). Da die im anliegenden Verzeichnisse aufgeführten juridischen und die wenigen am Ende erscheinenden medizinischen nebst ein paar andern Werke, meist rein theoretischen Gehaltes dem löbl. Stifte keinen beträchtlichen praktischen Nutzen gewähren zu können vermuthen lassen, hier aber an der Universität den h. Professoren zum Nachschlagen besonders in Bezug auf das römische Recht dessen Gehaltes die meisten sind ersprießliche dienste förderhin thun würden, so glaubt der gehorsamst Unterzeichnete anliegendes Verzeichniß ohne unbescheiden zu erscheinen mit der bitte begleiten zu dürfen die hochwürdige Vorstehung des löblichen Stiftes Neustift, wolle diese Werke der hiesigen Bibliothek gefälligst belassen und dagegen erwarten, daß der gehorsamt Unterzeichnete alle übrigen noch vorhandenen Werke (mit Ausnahme der schon im Jahre 1819 vom H. sel. Prälaten durch die hohe Landesstelle der hiesigen Bibliothek großmüthig cedirten typographischen Denkmählern des 15. Jahrhunderts in Bälde mit aller Gewissenhaftigkeit ausfolgen werde. Betreffend den so genannten Teyerdank erlaubt sich der gehorsamst Unterzeichnete noch die Bemerkung, daß der selbe bekanntlich nur einen affectations Werth habe und wirklich wie das Verzeichniß sagt defect ist, hier aber aus dem Grund behaltenswerth wäre, weil die hiesige Bibliothek noch einen hat der defect ist und beyde zusammen einen ganzen bilden. Die schönen Beyspiele der übrigen hochverehrlichen Stifte unserer Provinz die so gefällig waren zum Vortheile der öffentlich guten Sache viele ihrer Werke dazulassen (das lobliche Stift Stams hat die seinen hier befindlichen der Univers. Bibliothek samt und sonders geschenkt) und das vom h. sel. Prälaten des Stiftes Neustift durch Abtrettung der schon genannten typogr. Denkmahler erzeigte Wohlwollen des Stiftes Neustift lassen auch dermahl keine fehlbitte befürchten. Innsbruck, den 19. Mai 1833 (Innsbruck, ULB Tirol, Direktionsakt 1833, Nr. 114). In einem Nachtrag vom 25. Mai desselben Jahres zu jenem Schreiben ersucht Bibliothekar Bertholdi die Stiftsleitung nochmals um völlige Resignation für jene Bücher und Werke, die in dem vorigen Schreiben beigelegten Verzeichnis offenbar angemerkt wurden, um eine neue Stöhrung in der nun zu Stande zu bringenden Stellung und Signatur wie auch in die anzufertigenden neuen Cataloge zu vermeiden. Eine nächste Bitte, weitere Bücher der Universitätsbibliothek zu überlassen und diese in einem beigelegten Verzeichnis anzuzeigen (wie aus einem anderen Schreiben hervorgeht, ist dieses Verzeichnis sieben Bogen stark), folgt bereits am 14. Juni 1833; begründet wird dieses Ansuchen damit, weil viele von den zu übersendenden die einzigen waren, die wir in dieser Art oder aus diesem und jenem fache besassen (Innsbruck, ULB Tirol, Direktionsakt 1833, Nr. 126). Vgl. auch Brief des Bibliothekars Scherer an das Landesgubernium für Tirol und Vorarlberg vom 15. Juni 1833 (Innsbruck, ULB Tirol, Direktionsakt 1833, Nr. 127). Als Dank für die Zusage, der Universitätsbibliothek jene Bücher zu überlassen, welche dem Stifte minder nothwendig seyn dürften, schlägt Bibliothekar Scherer vor, den offensichtlich in Innsbruck beschädigten „Theuerdank“ auf Kosten der Landesstelle ergänzen zu lassen (Innsbruck, ULB Tirol, Direktionsakt 1833, Nr. 135, 8. Juli 1833). V. Tizzani, Reise nach Neustift zur Saecularfeier im Jahre 1842 und ein Ausflug auf den großen S. Bernhard. Terni 1846, 22. Y. Huguenin-Bergenat, Kulturgüter bei Staatensukzession. Die internationalen Verträge Österreichs nach dem Zerfall der österreichisch-ungarischen Monarchie im Spiegel des aktuellen Völkerrechts (Schriften zum Kulturgüterschutz / Cultural Property Studies). Berlin 2010, 113–122 (Art. 195 und 196). Die Stiftsbibliothek von Neustift 25 der Provinzen Italiens fortgebracht worden sind 82 zurückzugeben. Zur Umsetzung dieser Vorgabe sollten ehestmöglich Verhandlungen wegen Abschlusses eines gütlichen Uebereinkommens 83 auf genommen werden. Die Ausführungskonvention vom 4. Mai 1920 sah schließlich vor, dass Österreich „alle Kulturgüter samt den Archiven“ zurückerstatte, die „nach dem 1. Januar 1790 von den abge tretenen Gebieten nach Österreich gelangt waren“ und „nach ihrem Ursprung (origine) zum histo rischen und kulturellen Besitz (patrimoine historique et intellectuel) Italiens oder der abgetretenen Gebiete gehörten“.84 Gerade dieser Aspekt des Lokalbezugs und der historischen Bedeutung traf auch auf die Neustifter Handschriften zu. Im Namen der Neustifter Stiftsvorstehung richtete daher der Augustiner Chorherr Anselm Sparber am 5. November 1920 schließlich an den Direktor der Universitätsbibliothek Inns bruck die Frage, ob es nicht möglich wäre, die eine und andere historische Handschrift, welche für die Geschichte Neustifts von besonderer Bedeutung ist, speziell Cod. 931 […] zurückzuerhalten. Wie Ihnen wohl bekannt ist, finden sich in der Universitätsbibliothek ziemlich einige Handschriften, die von Neustift stammen, darunter auch historische. Von kompetenter italienischer Seite ist der Stiftsvorstehung nahegelegt worden, sie möchte die Rückgabe aller Handschriften fordern und betreiben. Doch davon will die Stiftsvorstehung nichts wissen. Vielmehr bin ich ersucht worden, den Vorschlag zu machen, ob nicht auf gütigem Wege etwas zu erreichen sei. Aus diesem und anderen Gründen gedenke ich um Neujahr hinauszufahren. Ich erlaube mir aber jetzt schon die Anfrage höflichst zu richten, ob irgendwelche Aussicht auf Erfolg vorhanden ist und ob Sie eine Eingabe in das österreichische Unterrichtsministerium befürworten würden. Es würde sich um Handschriften handeln, die für Neustift lokal geschichtlich von großem Werte oder fast unentbehrlich sind.85 Diese Bitte bzw. diese Ansprüche stießen allerdings auf wenig Entgegenkommen auf österreichischer Seite. Hans Tietze, dem als Referent für museale Angelegenheiten im Unterrichtsministerium die Verteidigung des österreichischen Kunst besitzes gegen Ansprüche von anderen Staaten oblag, teilte einem nicht näher benannten Kollegen mit, daß von den aus den abgetretenen Gebieten stammenden Gegenständen nur jene auszufolgen sind, die nach ihrem Ursprung (origine) zum historischen und kulturellen Besitz Italiens oder der an dasselbe abgetretene Provinzen gehören. Daher sei eine Abgabe der Inkunabeln nicht notwendig, denn unzweifelhaft ist für die Zugehörigkeit eines Inkunabels [!] der typographische Gesichtspunkt maßgebend. Das ist nach der ganzen Inkunabelforschung unzweideutig. Ganz genau dasselbe gilt von den Handschriften. Handschriften, die in Deutschland oder überhaupt außerhalb von Südtirol entstanden sind, können nicht beansprucht werden.86 Aus diesem Grund bat er als Unterlage für die weiteren Verhand lungen um eine Liste mit möglichst genauen Angaben. In einem weiteren Schreiben regte er die Möglichkeit eines Tausches an: Hier frage ich Sie nun, ob Sie es für diskutabel hielten, ein [!] ganz freien Ausgleich vorzuschlagen, z. B. für Ihre Handschrift 100 87, Ihre Handschrift 87 88 anzubieten. Es ist natürlich schwer zwei derartig unvergleichliche Dinge gegen einander abzuschätzen, zumal ja auch der Seneca alte Tiroler Beziehungen besitzt. Immerhin aber könnte es geltend gemacht werden, daß es sich hier um eine charakteristische italienische Arbeit handelt. Während Nr. 100 doch in der Tat die 82 83 84 85 86 87 88 Innsbruck, ULB Tirol, Direktionsakt 1920 ad Nr. 135, 2. Ebda 3. Huguenin-Bergenat, Kulturgüter (wie Anm. 81) 128. Innsbruck, ULB Tirol, Direktionsakt 1920, Nr. 135. Innsbruck, ULB Tirol, Direktionsakt 1921, Restitution, Fasz. 2 (Wien, 26. Februar 1921). Einen entsprechenden Hinweis betreffend die Abgabepflicht an Italien im Frühjahr 1921 findet sich auch in: Wien, Haus-, Hof- und Staatsarchiv, SB Registratur Archivbevollmächtigter, Zahl 28/1921. Freundlicher Hinweis David Fliri, Wien. Heute Neustift, Stiftsbibl., Cod. 100 („Posch-Missale“). Heute Innsbruck, ULB Tirol, Cod. 87. Diese Pergamenthandschrift enthält die „Tragoediae“ des Seneca und wurde Ende des 14. Jahrhunderts in Bologna geschrieben. Ein Bezug zu Neustift ist nicht gegeben; die Handschrift war im Besitz des Salzburger Fürsterzbischofs Wolf Dietrich von Reichenau (1559–1617), fand anschließend Eingang in die sogenannte Wappenturmbibliothek in Innsbruck und gelangte von dort in die ULB Tirol; vgl. Katalog ULB Tirol I (wie Anm. 100) 237–241 sowie Abb. 6 und 36. 26 Die Stiftsbibliothek von Neustift bedeutendste Tiroler Miniaturmalerei ist.89 Ob dieser Vorschlag dem Direktor des Ufficio regionale per i Monumenti, le Belle Arti e le Antichità (ab Dezember 1923 Soprintendenza) Giuseppe Gerola dann auch tatsächlich unterbreitet wurde, ist nicht bekannt. Er kam jedenfalls nicht zustande. Bereits zwei Wochen später folgte ein Schreiben Tietzes an eben diesen Kollegen, dass es ihm gelungen sei, den größten Teil der deutschen Handschriften und sämtliche Inkunabeln […] für Innsbruck zu retten. Die Italiener haben verzichtet auf die Nummern 59, 82, 164, 169, 170, 172, 175, 176, 596, 598, 842, 922, 157, 159, 763, 1015.90 Glücklicherweise befinden sich darunter die Rechts-Handschriften, auf die Sie ja das größte Gewicht gelegt haben. Dagegen ist es mir leider nicht gelungen, die Handschrift 100 frei zu bekommen, weil ja die Beziehungen zu Neustift zu offensichtliche sind.91 Eine Einigung mit dem italienischen Staat dürfte bereits kurze Zeit später erfolgt sein. Am 29. August desselben Jahres wurden die Handschriften entsprechend einem Auftrage des Unterrichtsamtes Ettore Modigliani (Soprinten dente alle Gallerie, ai Musei Medievali e Moderni e agli Oggetti d’arte di Milano) und Giuseppe Gerola übergeben92 und gelangten als Staatsbesitz an die Soprintendenza alle Belle Arti nach Trient.93 Fast acht Jahre sollte es dauern, ehe am 7. August 1929 schließlich 100 Handschriften a titolo di semplice deposito temporaneo nach Neustift zurückkehrten94; bereits zwei Jahre später wurde um eine Dauer leihgabe ersucht. Das Stift verpflichtete sich, sie mit größter Sorgfalt aufzubewahren und sie gegen 89 90 91 92 93 94 Innsbruck, ULB Tirol, Direktionsakt 1921, Restitution, Fasz. 3 (Wien, 3. März 1920 [korrigiert zu 21]). Diese hier genannten Handschriften werden bis heute in der ULB Tirol in Innsbruck aufbewahrt; vgl. die Liste im Anhang. Innsbruck, ULB Tirol, Direktionsakt 1921, Restitution, Fasz. 1 (Wien, 14. März 1921). Das Präsidium der Tiroler Landesregierung wurde am 22. März darüber informiert. Am 7. April teilte Ludwig Sprung, Direktor der Universitätsbibliothek Innsbruck, dem Prälaten von Neustift mit, dass von Seite der italienischen Regierung durch den Herrn Kommissär Dr. Gerola ursprünglich sämtliche aus Neustift stammenden Handschriften und Bücher beansprucht wurden, weil die italienischen Vertreter die Giltigkeit der Schenkung vom 18. Juli 1833 nicht anerkennen. Unserem Einspruche und den Bemühungen der österreichischen Vertreter in Wien ist es gelungen, aus diesem für uns äusserst wertvollen Bestande durch Verzicht der italienischen Komissäre alle Drucke und von den Handschriften die Nr. 59, 82, 157, 159, 170, 172, 175, 176, 596, 598, 763, 842, 922 & 1015, ausserdem die nach Herkunft zweifelhaften Nr. 111, 255, 324, 593, 607, 642, 692, 693, 717, 718, 723, 765, 799, 800, 829, 988 für unsere Anstalt zu retten. Alle übrigen, darunter auch alle auf Neustift selbst bezüglichen sollen an einem noch unbestimmten Termine abgeliefert werden. Es ist natürlich für das Kloster und das Land von grosser Wichtigkeit, dass diese Handschriften nicht verschleppt werden, sondern wieder im Kloster selbst oder wenigstens in einem deutschen Südtirolerort ihre Aufbewahrung finden (Neustift, Stiftsarchiv, ohne Sign., 7. April 1921). Schreiben des Direktors der Universitätsbibliothek Innsbruck Ludwig Sprung an den Prälaten von Neustift, 30. August 1921 (Neustift, Stiftsarchiv, ohne Sign.). In ebendiesem Schreiben äußert der Bibliotheksdirektor auch den Wunsch, dass es Ihren Bemühungen gelingen wird, diese Handschriften dem Kloster und damit dem deutschen Besitzstande Südtirols zu retten. Vgl. Schreiben des Propstes Bernhard Haller an das Commissariato Generale Civile, Uffizio Belle Arti, 5. September 1921 (Archiv des Amtes für Bau- und Kunstdenkmäler Bozen, Abteilung Denkmalpflege, Fasz. Neustift). Der Propst bat darum, die Handschriften nach Neustift zu überstellen. Neustift, Stiftsarchiv, ohne Sign., „Verbale“ (ohne Datum). Eine Kopie dieses Verbale war dem Schreiben vom 2. August 1929 (Oggetto: Deposito di manoscritti) beigelegt und musste unterschrieben zur Übergabe nach Trient mitgebracht werden (vgl. Neustift, Stiftsarchiv, ohne Sign.). Im Verbale sind 99 Handschriften maschinschriftlich aufgelistet, Cod. 1015 ist handschriftlich ergänzt. Bereits seit Juli stand die Soprintendenza mit dem Konvent in regem Austausch, um die Voraussetzungen für eine Übergabe zu schaffen. Am 23. Juli wurde nach Trient berichtet, dass in jüngster Zeit nicht weniger als 10.000 Lire für die Bibliothek aufgewendet wurden und u. a. das Dach der Bibliothek erneuert worden sei. Diesem Entschluss sind Überlegungen vorausgegangen, die Handschriften im Staatsarchiv Bozen oder im Diözesanmuseum in Brixen unterzubringen. Propst Bernhard Haller wies in mehreren Schreiben an Giuseppe Gerola darauf hin, dass Neustift über geeignete Räumlichkeiten zur Aufbewahrung der Handschriften verfüge und aufgrund anderer im Stift vorhandener Handschriften eine Zusammenführung sinnvoll sei. Zudem unterstrich er die Bedeutung der Handschriften für die Geschichte des Stiftes. Warum nicht alle an der Universitätsbibliothek Innsbruck aufbewahrten Handschriften mit eindeutig Neustifter Provenienz abgegeben werden mussten, sondern das Unterrichtsministerium nicht nur auf die oben genannten Signaturen, sondern auch noch auf weitere Handschriften verzichtete (vgl. Liste im Anhang), bleibt offen. Die Stiftsbibliothek von Neustift 27 Diebstahl, Feuchtigkeit und Feuer zu schützen. Eine weitere Neustifter Handschrift, Cod. 142, wurde erst 1931 definitiv von Trient nach Neustift überstellt95; Cod. 1015, dessen Signatur dem Verzeichnis zu einem späteren Zeitpunkt handschriftlich hinzugefügt wurde, ist in Innsbruck verblieben.96 Genau diese Handschriften stehen im Zentrum des vorliegenden Katalogs; ausgenommen sind lediglich die Archivhandschriften, die offensichtlich 1807 ebenfalls nach Innsbruck gebracht worden waren und dort eine Bibliothekssignatur erhalten hatten, sowie Handschriften aus späteren Jahrhunder ten. Dazu zählen: Cod. 121: Cod. 501: Cod. 503: Proprium sanctorum … ab norma Romani Breviarii accomodatum97 [17. Jh.] Urbario [14. Jh.] Calendarium [Kalendar mit Eintragung von Jahrtagen, Märkten, Gerichtstagen und diversen Anweisungen aus dem Jahr 1511 mit späteren Nachträgen] Cod. 643: Partecipazione di morte di P. P. [Bitte des Klosters Neustift an die konföderierten Klöster, verstorbene Priester in den Nekrolog einzutragen; 45 Klöster haben ihr Willfahren dieser Bitte vermerkt, 1618] Cod. 689: Johanis Stroza Oratio [17. Jh.] Cod. 836: Rendite di Novacella 1441 [Urbar, 13. Jh. mit späteren Nachträgen] Cod. 836: Urbario [15. Jh. mit späteren Nachträgen] Cod. 839: Urbario [Abschrift der Privilegien Neustifts, Hinweise zu Jahrtagen von Stiftern, Stiftungen, 15. Jh.] Cod. 845: Nomina Praepositorum novacellensium [Übersicht über die Pröpste von Neustift von 1143 bis 1689 mit Darstellung ihrer Wappen, 17. Jh.] Cod. 857: Relation [Sammlung von Akten aus der Zeit Propst Augustin Schabls, 1569–1571, zusammengestellt 1572] Cod. 924: Urbario [15. Jh.] Cod. 927: Officium [Urbar, 14. Jh.] Cod. 930: Elenco degli Abati di Novacella [Übersicht über die Pröpste von Neustift, 18. Jh.] Cod. 931: Epitome de gestis Praelatorum Novacellensium [umfangreiches Werk zu den einzelnen Pröpsten, Propst Fortunat Troyer gewidmet, 1693] Cod. 1012: Regula [Bulla papae Nicolai IV. „Supra Montem“, 1. Viertel 16. Jh.] Zusätzlich zu den 85 aus Innsbruck wieder nach Neustift zurückgebrachten Bibliotheks handschriften sind im vorliegenden Katalog weitere sieben Codices berücksichtigt, die zu Beginn des 19. Jahrhunderts nicht nach Innsbruck gebracht worden sind. Hierzu zählen drei Handschriften ohne Signatur: das sogenannte Neustifter Wappenbuch und die zwei großformatigen Gradualbände aus dem 15. Jahrhundert; zwei Handschriften mit einer Bibliothekssignatur (Numerus currens, wohl aus dem 95 96 97 Die Signatur Cod. 142 ist in der dem unter Anm. 94 erwähnten „Verbale“ beigelegten Liste „Codici del Convento di Novacella“ handschriftlich ergänzt. Die Handschrift war bereits zu einem früheren Zeitpunkt als Leihgabe nach Neustift geschickt worden. Anselm Sparber verlieh sie an Leo Santifaller, der sie schließlich beim Staatsarchiv in Bozen zurückgab, vgl. Schriftwechsel zwischen Giuseppe Gerola und Antonio Zieger, Direktor des Staatsarchivs von Bozen, 23. und 24. März 1931. Im März 1931 gelangte sie wieder an die Soprintendenza all’arte medioevale e moderna Trento; Gerola sicherte daraufhin dem Stift zu, sie bei nächster Gelegenheit und zu den gleichen Bedingungen wie die anderen nach Neustift zu bringen, vgl. Schreiben vom 31. März 1931 (Archiv des Amtes für Bau- und Kunstdenkmäler Bozen, Abteilung Denkmalpflege, Fasz. Neustift). Innsbruck, ULB Tirol, Cod. 1015 (Cosmographia. Scriptores geographiae. Neustift 1509); vgl. Katalog ULB Tirol X (wie Anm. 100) 188–189. Die Bezeichnungen sind allesamt dem Anhang des „Verbale“ entnommen (siehe Anm. 94), in eckigen Klammern wurden Informationen, vorwiegend entnommen aus dem Zettelkatalog „Die historischen Handschriften des Archivs bzw. der Bibliothek des Augustiner-Chorherrenstiftes Neustift“ (online verfügbar unter http://www.ksbm.oeaw.ac.at/kataloge/IT/5000/histHss/histHss.pdf), ergänzt. 28 Die Stiftsbibliothek von Neustift späten 19. Jahrhundert stammend): Sign. 14820 (Rituale), Sign. 15063 (Antiphonarium); sowie zwei Handschriften, die erst in jüngster Zeit mit Nummern versehen wurden: ein Graduale und Antiphona rium, das ursprünglich die Bibliothekssignatur Sign. 1481998 aufwies und um 1970 die Bezeichnung Nr. 139 erhielt, sowie ein weiteres Graduale und Antiphonarium, geschrieben von der gleichen Hand wie Nr. 139, das erst um das Jahr 2000 mit Nr. 940 versehen wurde.99 Keine Berücksichtigung finden hingegen jene Neustifter Handschriften, die heute an der ULB Tirol aufbewahrt werden; vgl. hierzu Anhang. Sie wurden bereits im Rahmen des Projektes zur Er schließung des dortigen Handschriftenbestandes bearbeitet.100 98 99 100 Für die Freilegung der Bibliothekssignatur aus der Zeit um 1910 und weitere Auskünfte geht ein Dank an Stiftsarchivar Simon Terzer. Vgl. hierzu „Übersicht über den Handschriftenbestand“, Abschnitt „Signaturen“. Beim österreichischen Bibliothekartag in Eisenstadt 1972 brach der damalige Direktor der Handschriften- und Inkunabelsammlung der ÖNB Otto Mazal (1932–2008) eine Lanze für den Beginn von Generalkatalogen nach dem Vorbild der Deutschen Forschungsgemeinschaft (DFG) und rief eine Arbeitsgruppe ins Leben, die die genauen Beschreibungsrichtlinien festsetzen sollte. Nach intensiven Diskussionen und Arbeitstagungen in Kremsmünster (1973) und Zwettl (1974) lagen die Richtlinien 1975 in gedruckter Form vor; bereits kurze Zeit später wurde mit der entsprechenden Erschließung an der ULB Tirol unter der Leitung von Walter Neuhauser begonnen. Seither wurden, dem Numerus currens folgend, die gesamten dort aufbewahrten Handschriften nach diesen Richtlinien erschlossen. Die Beschreibungen liegen in insgesamt zehn Bänden vor: Katalog der Handschriften der Universitätsbibliothek bzw. Universitäts- und Landesbibliothek Tirol in Innsbruck, unter der Leitung von W. Neuhauser. T. 1: Cod. 1–100, bearb. von W. Neuhauser; T. 2: Cod. 101–200, bearb. von W. Neuhauser; T. 3: Cod. 201–300, bearb. von G. Kompatscher; T. 4: Cod. 301– 400, bearb. von W. Neuhauser, L. Šubarić; T. 5: Cod. 401–500, bearb. von D. Mairhofer, W. Neuhauser, M. Rossini, C. Schretter; T. 6: Cod. 501–600, bearb. von D. Mairhofer, W. Neuhauser, C. Schretter, U. Stampfer; T. 7: Cod. 601–700, bearb. von P. Ausserlechner, W. Neuhauser, C. Schretter, U. Stampfer; T. 8: Cod. 701–800, bearb. von P. Ausserlechner, W. Neuhauser, A. Ohlenschläger, C. Schretter, U. Stampfer; T. 9: Cod. 801–950, bearb. von P. Ausserlechner, H. Gritsch, P. Kennel, W. Neuhauser, A. Ohlenschläger, C. Schretter-Picker, U. Stampfer; T. 10: Cod. 951–1198, bearb. von H. Gritsch, P. Kennel, W. Neuhauser, R. Neyer, A. Ohlenschläger, A. Pinter, C. Schretter-Picker (Denkschriften der ÖAW, phil.-hist. Kl. 192, 214, 271, 327, 365, 375, 414, 456, 479, 489 = Veröffentlichungen der Kommission für Schrift- und Buchwesen des Mittelalters II, 4, 1–10). Wien 1987, 1991, 1999, 2005, 2008, 2009, 2011, 2014, 2015, 2017. Seit 2009 werden die Ergebnisse zudem auch in die von der ÖAW betreuten Datenbank www.manuscripta.at eingepflegt. Dort sind auch die in Neustift verwahrten Handschriften zu finden, sodass nunmehr zumindest eine virtuelle Zusammenführung des einstigen Bestandes gewährleistet ist. Zum Projekt der Handschriftenerschließung in Tirol vgl. u. a. W. Neuhauser, Generalkataloge zu Handschriften österreichischer Bibliotheken, in: K. Niedermair (Hrsg.), Die neue Bibliothek. Anspruch und Wirklichkeit. 31. Österreichischer Bibliothekartag Innsbruck 2011 (Schriften der Vereinigung Österreichischer Bibliothekarinnen und Bibliothekare 11). Graz, Feldkirch 2012, 38–43. 29 2. Übersicht über den Handschriftenbestand Ursula Stampfer Insgesamt werden heute in Neustift 92 mittelalterliche Bibliothekshandschriften101 aufbewahrt. Zudem sind noch zahlreiche Archiv- sowie neuzeitliche Handschriften erhalten. Die einzelnen Bestände sind weder von ihrer Aufstellung noch hinsichtlich ihrer Signaturen klar voneinander abgegrenzt; jeweils mehrere unterschiedliche Signatursysteme kommen zum Tragen. An der ULB Tirol werden weitere 40 Handschriften aufbewahrt, die zu Beginn des 19. Jahr hunderts von Neustift nach Innsbruck gebracht wurden (siehe oben). Bei zusätzlich 37 Handschriften kann nicht eindeutig festgestellt werden, ob sie aus Neustift nach Innsbruck gelangten. Für zwei andere Handschriften kann hingegen davon ausgegangen werden, dass sie einst zur Neustifter Bibliothek gehörten, aber bereits zu einem früheren Zeitpunkt ausgeschieden wurden. Im Folgenden richtet sich der Fokus auf den heute in der Stiftsbibliothek Neustift aufbewahrten Bestand; eine Zusammenschau des gesamten ursprünglichen Neustifter Bestandes stellt derzeit noch ein Desiderat dar. 2.1 Signaturen Nach der vorübergehenden Klosteraufhebung wurden nicht nur mittelalterliche Bibliothekshandschrif ten,sondern auch Teile der Archiv- und neuzeitlichen Handschriften nach Innsbruck gebracht und der Universitätsbibliothek einverleibt. All diese tragen heute jene Signaturen, die sie im Zuge der Neuauf stellung des Buchbestandes im 19. Jahrhundert ebendort erhielten. Die zahlreichen Handschriften, die aufgrund der kirchenpolitischen Maßnahmen im ausgehenden 18. und beginnenden 19. Jahrhundert aus unterschiedlichen Klöstern nach Innsbruck gebracht worden waren102, wurden hier aber nicht nach Provenienz aufgestellt und sind deshalb auf den gesamten Handschriftenbestand verteilt. Unter Direktor Martin Scherer (1832–1857) erhielten alle an der Universitätsbibliothek Innsbruck aufbewahrten Drucke und Handschriften eine mehrteilige Standortsignatur; zudem wurde für die Handschriften eine laufende Inventarnummer vergeben. Während die Standortsignaturen unter Friedrich Leithe (1868– 1874) abgeändert wurden, behielten die Handschriften den Numerus currens bei, der sowohl am Spie gel des Vorderdeckels als auch auf einem gelben Papierschildchen am Rücken vermerkt wurde.103 Daher reichen die Signaturnummern der heute in Neustift aufbewahrten Handschriften von 4 bis 1012; 101 102 103 Als Grenze zwischen mittelalterlichen und neuzeitlichen Handschriften wurde die Mitte des 16. Jahrhunderts festgesetzt, da der überwiegende Teil der Handschriften bis zu jenem Zeitpunkt mittelalterlichen Charakter aufweist. Zur Geschichte der Handschriftensammlung an der ULB Tirol siehe insbes. W. Neuhauser, C. SchretterPicker, P. Zerlauth, P. Kennel, Das Alte Buch an der ULB Tirol. Erbe und Auftrag, in: K. Niedermair, D. Schuler (Hrsg.), Die Bibliothek in der Zukunft: regional – global. Lesen, Studieren und Forschen im Wandel. Festschrift für Hofrat Dr. Martin Wieser. Innsbruck 2015, 245–276; Neuhauser, Geschichte (wie Anm. 72); Hittmair, Geschichte (wie Anm. 72). Zu diesen Signaturen vgl. Katalog ULB Tirol I (wie Anm. 100) 20–21. 30 Übersicht über den Handschriftenbestand die durch die Abgabe der Bücher an Italien nach dem Ersten Weltkrieg hervorgerufenen Lücken in der Signaturenreihe der Innsbrucker Handschriften wurden nicht geschlossen.104 Drei Handschriften, Sign. 14819, 14820 und 15063, weisen eine Signatur in Tinte auf natur farbenem Papierschildchen am unteren Rücken auf. Diese Signaturen stammen aus der Zeit um 1910, als sämtliche im barocken Bibliothekssaal aufgestellten Bücher mit einem Numerus currens versehen wurden.105 Sie zeigen sehr anschaulich auf, dass einst Drucke und Handschriften – zumindest in Teilen – gemischt eingereiht waren.106 Aus wesentlich jüngerer Zeit stammen die Signatur Nr. 940 sowie die Neu-Nummerierung von Sign. 14819 mit Nr. 139. Sowohl 940 als auch 139 würden sich zwar gut in die Innsbrucker Signaturen reihe einfügen, doch sind sie dort bereits anderweitig vergeben.107 Warum gerade diese beiden N ummern für die von derselben Hand geschriebenen Handschriften gewählt wurden, bleibt offen. Die Nummer 139 wurde vom Stiftsbibliothekar H. Martin Peintner vor 1972 in Rot auf einem weißen Etikett am unteren Buchrücken festgehalten.108 Das Signaturschild mit der Nummer 940 in schwarzer Tinte stammt aus der Zeit um 2000 und wurde von H. Theobald Innerhofer auf den wohl kurz vorher neu gestalteten Bibliotheksband aufgeklebt. Spuren einer älteren Signatur sind hier keine auszumachen. Die beiden großformatigen Gradualbände aus der Mitte des 15. Jahrhunderts sowie das sogenannte Neustifter Wappenbuch weisen hingegen gar keine Signaturen auf. Der Grund hierfür liegt möglicher weise darin, dass sie ursprünglich nicht in der Bibliothek aufbewahrt wurden. Für die Gradualbände, in denen Bleistiftnotizen auf eine Verwendung bis ins späte 19./frühe 20. Jahrhundert schließen lassen, darf wohl die Kirche bzw. die Sakristei als Aufbewahrungsort angenommen werden. Von einer Vergabe von neuen, alle mittelalterlichen Handschriften zu einer Einheit zusammen fassenden Signaturen wurde aufgrund der in bisherigen Publikationen üblichen Verwendung der „Inns brucker Signaturen“ abgesehen. Auch die in jüngerer Zeit in Neustift vergebenen Signaturen wurden beibehalten. 2.2 Fachgebiete/Inhalte Erwartungsgemäß sind die meisten überlieferten Werke theologischen Inhalts (61): Missalien, Gra dualien, Ritualien und Antiphonarien, Bibeln bzw. Bibelauszüge, Psalmenkommentare, Gebetbücher sowie Ordensliteratur, d. h. Ordensregeln und deren Auslegungen, Berichte sowie Texte zur Ordensund Hausgeschichte, zahlreiche Predigtsammlungen, homiletische Traktate, Texte von Kirchenvätern und Schriften aszetischen sowie dogmatischen Inhalts. 104 105 106 107 108 Vgl. hierzu Katalog ULB Tirol I–X (siehe Anm. 100): Bei allen abgetretenen Handschriften bzw. deren Signaturen wurde ein entsprechender Eintrag vorgenommen. Zu diesen Signaturen ist ein Zettelkatalog in Karteikartenformat erhalten, geschrieben vorrangig von zwei Händen mit einzelnen späteren Ergänzungen. Hand 2 hat mehrere Karteikarten am unteren Blattrand datiert (Juli 1911). Heute befinden sich im Bibliothekssaal ausschließlich Drucke. Auch die Karteikarten mit Einträgen zu handgeschriebenen Büchern werden heute separat aufbewahrt. Wo sich die zahlreichen, vorwiegend aus dem 17. und 18. Jahrhundert stammenden hierin genannten Choralhandschriften befinden, ist nicht bekannt. Das Musikarchiv war zum Zeitpunkt der Recherchearbeit nicht zugänglich. Innsbruck, ULB Tirol, Cod. 139: Thomas de Aquino, Albertus de Diessen, entstanden in Süddeutschland und Schnals (?) um 1425/35 (Letztbesitzer: Schnals); vgl. Katalog ULB Tirol II (wie Anm. 100) 108–109. Innsbruck, ULB Tirol, Cod. 940: Iacobus de Voragine, Entstehungsort unbestimmt, 14. (u. a. 1372) und 15. Jh. (Letztbesitzer: Schnals); vgl. Katalog ULB Tirol IX (wie Anm. 100) 351–354. Die Signatur stimmt mit der Nummer im maschinschriftlichen „Verzeichnis der Handschriften. Aus der Bibliothek des Klosters Neustift b. Brixen. Zusammengestellt von Prof. Martin Peintner – Bibliothekar. 1972“ überein. Das Verzeichnis listet insgesamt 504 Handschriften auf; der Großteil stammt aus dem 18. und 19. Jahrhundert, einige datieren in die erste Hälfte des 20. Jahrhunderts. Übersicht über den Handschriftenbestand 31 Die restlichen Handschriften bieten einen Querschnitt durch den Bildungskanon jener Zeit und zeigen einen breit gefächerten Bibliotheksbestand, ohne besondere Schwerpunkte. Juristische bzw. insbesondere kanonistische Texte (9), vielfach begleitet von bekannten Glossen, sind ebenso vorhanden wie medizinische und naturwissenschaftliche Abhandlungen (6). Die schöngeistige Literatur ist trotz des seit dem 13. Jahrhundert in Neustift belegten Schulbetriebs eher spärlich vertreten (12), Grammatik en u. ä. fehlen zur Gänze. Für den Unterricht vorgesehen waren möglicherweise auch die zwei in einem Band vereinten musiktheoretischen Traktate samt Schemata zu Tonschlüssen, denen astronomische Texte beigefügt sind. Drei Handschriften sind schließlich den Historica zuzuordnen, darunter auch ein Wappenbuch. Ein ergänzender grober Überblick über die Inhalte der heute in Innsbruck aufbewahrten Neustifter Handschriften findet sich im Anhang. 2.3 Zeitliche Verteilung Die Handschriften datieren vom 10. Jahrhundert bis ins Jahr 1524 bzw. 1526: Während die Schrift dieses jüngsten im vorliegenden Katalog berücksichtigten Codex noch eindeutig der mittelalterlichen Tradition verpflichtet ist, weist die Buchmalerei bereits in die Renaissance. So beschließt diese Hand schrift das mittelalterliche Skriptorium von Neustift. Auffallend ist die geringe Anzahl an frühen Handschriften – auch die an der ULB Tirol aufbewahr ten Neustifter Handschriften datieren von der zweiten Hälfte des 14. bis ins 16. Jahrhundert; sie ist aber angesichts der zahlreich erhaltenen Fragmente109 zu relativieren. Es ist vielmehr davon auszugehen, dass die Buchbestände aus der Gründungszeit des Klosters lediglich nicht die Zeiten überdauert haben und durch neuere Handschriften und später Inkunabeln bzw. gedruckte Werke ersetzt werden konnten. Hierzu zählen u. a. die im Mittelalter weit verbreiteten Lehrbücher wie die „Ars maior“ und die „Ars minor“ des Aelius Donatus oder das „Doctrinale“ des Alexander de Villa Dei. Reste von Letzterem sind als Fälze in Cod. 52 wiederverwendet worden. Der überwiegende Teil der erhalten gebliebenen Handschriften stammt aus dem 15. Jahrhundert. Sie legen einerseits ein anschauliches Zeugnis über die wirtschaftliche und kulturelle Blütezeit des Stiftes in jener Zeit ab, machen andererseits aber auch deutlich, dass auch nach der Erfindung des Buch druckes weiterhin Bücher handschriftlich angefertigt wurden. Der gleichzeitige Ankauf von mehr als 800 Inkunabeln zeigt auf, dass anstelle eines Entweder-oder vielmehr ein Sowohl-als-auch der Realität entsprach. 10. Jh. 12. Jh. 12./13. Jh. 13. Jh. 13./14. Jh. 14. Jh. 14./15. Jh. 109 495 154, 282, 315, 327, 360, 483 20, 293, 485 19, 86, 107 156, 171, 605 7, 52 (u. a. dat. 1396), 31, 99, 161, 162, 168, 253, 308, 532, 606 83 (dat. 1400), 158, 163, 182 (dat. 1400), 183 Zahlreiche abgelöste Fragmente werden in einer eigenen Mappe im Stift Neustift aufbewahrt; andere wurden als Makulatureinbände für Drucke, aber auch für Archivhandschriften und Placita wiederverwendet. Im Rahmen eines Projektes der Freien Universität Bozen unter der Leitung von Giulia Gabrielli werden die Neustifter Fragmente in der Stiftsbibliothek und an der ULB Tirol von Gionata Brusa und Giovanni Varelli erfasst. Die abgelösten Neustifter Fragmente an der ULB Tirol fanden in einem vom Jubiläumsfonds der Österreichischen Nationalbank geförderten Projekt („Die abgelösten Fragmente der ULB Tirol und ihre digitale Erschließung“, 2018–2020) unter der Leitung von Martin Wagendorfer und der Mitarbeit von Claudia Sojer (Universität Innsbruck, München) Berücksichtigung. Die Ergebnisse werden über https://fragmentarium.ms bereitgestellt. Vgl. die weiterführenden Informationen auf der Homepage der Abteilung für Sondersammlungen der ULB Tirol: https://www.uibk.ac.at/ulb/sondersammlungen/2.1.1.2-fragmente.html. 32 Übersicht über den Handschriftenbestand 15. Jh. 15./16. Jh. 16. Jh. 4, 38, 53 (u. a. dat. 1452), 76, 77 (dat. 1465), 78, 106, 123, 125, 142 (u. a. dat. 1459), 145 (dat. 1427), 146, 153, 155 (u. a. dat. 1463), 160, 165 (u. a. dat. 1407), 166, 167 (dat. 1419), 173, 174 (u. a. dat. 1472), 177, 181, 196, 200, 204, 217, 252, 353, 397, 440, 447 (dat. 1414), 453, 550 (u. a. dat. 1465), 578 (dat. 1480), 579 (dat. 1491), 597, 614 (u. a. dat. 1419), 617 (u. a. dat. 1455, 1467), 654 (dat. 1496), 748, 754, 756 (dat. 1479), 780, Sign. 15063, Nr. 139 (olim Sign. 14819) (u. a. dat. 1495, 1496), Nr. 940 (u. a. dat. 1490, 1493), Neustifter Wappenbuch, Graduale Neocellense I (dat. 1442), Graduale Neocellense II 190, 755 85, 100 (dat. 1524–1526), 142 (u. a. dat. 1521), 194 (dat. 1507), 405 (dat. 1501), 691, 694, 726 (dat. 1523), 776 (u. a. dat. 1502), Sign. 14820 2.4 Entstehungsorte Die Entstehungsorte der Handschriften reichen von Nordfrankreich bis nach Oberitalien, wobei der Schwerpunkt auf dem Süddeutschen Raum im Allgemeinen und Tirol im Speziellen liegt. Elf Hand schriften weisen eindeutige Hinweise auf eine Herstellung in Neustift selbst auf, bei weiteren acht kann diese mit großer Wahrscheinlichkeit angenommen werden, drei andere Handschriften lassen eine Produktion im stiftseigenen Skriptorium vermuten. Zusätzlich dürften sieben Handschriften im Raum Brixen bzw. im Umfeld des Stiftes entstanden sein, so z. B. jene liturgischen Bücher, die eindeutig auf eine Verwendung in einer der inkorporierten Pfarreien Neustifts schließen lassen. Dazu zählt u. a. das Teilmissale aus dem 12. Jahrhundert (Cod. 154), das aufgrund der Hervorhebung des Festes der hl. Margaretha (in Verbindung mit Matthias und Matthäus) nach Margen bei Terenten weist, oder das Graduale und Antiphonarium aus dem späten 15. Jahrhundert (Nr. 940), das in der Kirche St. Ulrich in Ilstern (Pfarrei St. Sigmund/Kiens) verwendet worden sein dürfte. Nicht zuletzt dank der genauen Analyse der Wasserzeichen konnten mehrere Handschriften nach Wien bzw. in den Wiener Raum lokalisiert werden. Seit jeher bestanden zwischen Kloster Neustift und dem Augustiner Chorherrenstift Klosterneuburg bei Wien enge Beziehungen, die sich auch auf den Bücherbesitz auswirkten.110 Zudem trug wohl auch das Studium einiger Neustifter Chorherren an der 1365 gegründeten Universität Wien hierzu bei111: Sie dürften Bücher in Wien kopiert oder erworben und schließlich nach Neustift mitgebracht haben. Bei diesen Büchern handelt es sich vorwiegend um auf Papier geschriebene Gebrauchshandschriften, die kaum Buchschmuck aufweisen. Sorgfältiger und aufwändiger gestaltet sind hingegen jene Handschriften, die wohl im Umfeld der Universität Bologna entstanden sind. Eine besondere Erwähnung verdient hierbei Cod. 217 („Satyrae“ des Iuvenal und des Persius Flaccus), der durchgehend palimpsestiert ist: Mithilfe einer Quarzlampe konnte ein ursprüng lich quer zum heutigen Schriftspiegel eingetragenes formularartiges Register festgestellt werden, das wohl einem stationarius zur Dokumentation des Verleihs der Pecien diente. Die Tradition solcher Pecienhandschriften, d. h. unter Aufsicht von einer Kommission von Universitätsprofessoren (peciarii) hergestellte und von der Universität als Lehrmittel anerkannte „Normexemplare“ in ungebundenen Lagen, die anschließend von stationarii zum Zweck weiterer Abschriften verliehen wurden, ist vor wiegend für die Universitäten Paris und Bologna belegt.112 110 111 112 W. Neuhauser, Buchgeschichtliche Beziehungen zwischen dem Wiener Raum und Tirol im Mittelalter, in: C. Schretter, P. Zerlauth (Hrsg.), In libris. Beiträge zur Buch- und Bibliotheksgeschichte Tirols von Walter Neuhauser (Schlern-Schriften 351). Innsbruck 2010, 405–424. Neuhauser, Wissenschaftspflege (wie Anm. 23) 93–94. Bei L. Santifaller, Verzeichnis der Deutschtiroler an der Wiener Universität im Mittelalter. Tiroler Heimat N.F. 1 (1928) 258–272 werden nur drei Studenten aus Neu stift angeführt (260); aus Brixen studierten zwischen 1365 und 1499 nicht weniger als 116 Studenten in Wien. G. Murano, Opere diffuse per exemplar e pecia (Textes et Études du Moyen Âge 29). Turnhout 2005; K. Christ, Petia. Ein Kapitel mittelalterlicher Buchgeschichte. Zeitschrift für Bibliothekswesen 55 (1938) 1–44; J. Destrez, La „pecia“ dans les manuscrits universitaires du XIIIe et du XIVe siècle, Paris 1935. Übersicht über den Handschriftenbestand 33 2.5 Schriften und Schreiberhände Den unterschiedlichen Entstehungszeiten und -orten sowie den diversen Verwendungszwecken ent sprechend lassen sich in den vorliegenden Handschriften zahlreiche verschiedene Schriftarten nach weisen.113 Nur wenige Schreiberhände sind namentlich bekannt. Aus dem ersten Viertel des 15. Jahrhunderts begegnen der nicht näher fassbare Nicolaus Frechen, Hainricus Chirchmair, der möglicherweise aus dem Benediktinerkloster Mallersdorf in der Diözese Regensburg stammt (de Malhenstorff), sowie der kaiserliche Notar Albert Weinkern aus Rosenheim, der um 1420 offensichtlich im Tiroler Raum wirkte. Von ihm stammt auch die Handschrift Brixen, Priesterseminarbibliothek, Cod. D.19 (dat. 1421) sowie München, BSB, Clm 8375 (dat. Innsbruck 1418). Zudem ist er als notarius publicus 1426 in Stift Wilten nachgewiesen.114 In der Mitte des Jahrhunderts schrieb der bereits erwähnte Augustiner Chor herr Friedrich Zollner das umfangreiche Graduale, wie er selbst im Kolophon des ersten Bandes fest hielt. Möglicherweise wies ebendieser Zollner auch in der zweiten Hälfte des 15. und zu Beginn des 16. Jahrhunderts tätige Neustifter Chorherren wie Erasmus Lämpl, Stephan Stet(t)ner und Franciscus Prensteiner in die Kunst des Schreibens ein. Erwähnung verdienen auch Georg Hölzl aus Mattighofen im oberösterreichischen Innviertel, der im Auftrag des Stadtrichters und bischöflichen Zöllners in Klau sen Kaspar Neuhauser 1496 ein zweibändiges Gebetbuch schrieb, sowie der aus Passau stammende Heinrich Edling(er). Er ist als Schreiber zweier Gradualien und Antiphonarien aus der Zeit um 1495/96 belegt, die schließlich in zwei inkorporierten Stiftspfarreien Verwendung fanden. 2.6 Beschreibstoffe 42 der insgesamt 92 Handschriften sind auf Pergament geschrieben, 48 auf Papier und zwei Codices sind Mischhandschriften. Das Pergament ist sehr unterschiedlich, teilweise sehr dick, teilweise ist es sehr fein bearbeitet und geglättet. Bei unterschiedlicher Bearbeitung der beiden Seiten – Kennzeichen des sogenannten südlichen Pergaments115 – wurde beim Binden immer sorgfältig auf eine regelmäßige Abfolge geachtet. Besonders bei den älteren Handschriften sind vermehrt auch Knochenlöcher und Nahtstellen festzustellen. Die Frage, ob das bei den Handschriften verwendete Pergament teilweise auch in Neustift selbst hergestellt wurde, muss offenbleiben. In der Mitte des 13. Jahrhunderts ist ein erster pergamenarius in Brixen belegt.116 Sämtliche Papierhandschriften wurden von Maria Stieglecker auf ihre Wasserzeichen hin genau untersucht (siehe Abschnitt 3.1), woraus sich wertvolle Hinweise zur Lokalisierung und Datierung er gaben. 113 114 115 116 Zur Entwicklung der Schrift im Brixner Raum von der Karolingischen Schrift bis in die zweite Hälfte des 16. Jahrhunderts mit Beispielen aus Neustift siehe L. Santifaller, Bozner Schreibschriften der Neuzeit 1500–1851. Beiträge zur Paläographie (Schriften des Instituts für Grenz- und Auslanddeutschtum an der Universität Marburg 7). Jena 1930, 19–36. Mairhofer, Urkundenbuch (wie Anm. 7) 514. Beim „nördlichen Pergament“ wurden hingegen die Haar- und Fleischseite gleichmäßig bearbeitet. Zu beiden Pergamenten vgl. L. Santifaller, Beiträge zur Geschichte der Beschreibstoffe im Mittelalter. Mit besonderer Berücksichtigung der päpstlichen Kurie, T. 1: Untersuchungen (Mitteilungen des Instituts für Österreichische Geschichtsforschung, Erg.-Bd. 16, 1). Graz, Köln 1953, 80–82; Ders., Vom Schrift- und Schreibwesen unserer Heimat im Altertum und im Mittelalter. Der Schlern 13 (1932) 178–191, hier 187; Ders., Über südliches und nördliches Pergament. Der Schlern 13 (1932) 458– 463. Santifaller, Vom Schrift- und Schreibwesen unserer Heimat (wie Anm. 115) 187. 34 Übersicht über den Handschriftenbestand 2.7 Sprachliche Verteilung Von den 92 in Neustift befindlichen mittelalterlichen Handschriften ist lediglich in einer einzigen ein Teil in deutscher Sprache verfasst. Cod. 146 umfasst auf den ersten fünf Blättern die Regel des hl. Augustinus auf Deutsch, die mitten in Kapitel 7 abbricht; anschließend folgen zwei verschiedene Auslegungen und Erläuterungen der Ordensregel in lateinischer Sprache. Alle anderen Werke sind zur Gänze in lateinischer Sprache geschrieben. Auch unter den in Innsbruck aufbewahrten Neustifter Handschriften finden sich nur wenige volkssprachliche Texte, darunter ein „Spiegel deutscher Leute“ (Innsbruck, ULB Tirol, Cod. 922) und zwei „Schwabenspiegel“ (Innsbruck, ULB Tirol, Cod. 169 und 842), von denen einer möglicherweise in Neustift selbst um 1410/20 abgeschrieben wurde. Die zur selben Zeit möglicherweise ebenso in Neustift entstandene Sammlung an Auszügen und Sentenzen aus den „Pluemen der Tugend“ des Hans Vintler (Innsbruck, ULB Tirol, Cod. 961) für Predigtzwecke macht deutlich, dass die deutsche Sprache auch im geistlichen Umfeld zunehmend an Bedeutung ge wann. Dasselbe Format und denselben Einband weist auch die sogenannte „Innsbrucker-Neustifter Spielehandschrift“ (Innsbruck, ULB Tirol, Cod. 960, dat. 1391) auf; sie enthält ein Maria-Himmel fahrts-Spiel, ein Osterspiel und ein Fronleichnamsspiel in Deutsch und bezeugt die lange Tradition der geistlichen Spiele in Neustift. Zumal aber in diesen Texten keine tirolischen Spracheigenheiten und -färbungen zu erkennen sind, ist nicht anzunehmen, dass die Handschrift auch vor Ort entstanden ist.117 Aufgrund der eingetragenen Todesnotiz zu Oswald von Wolkenstein 1445 wurde sie aber spätestens seit damals im Stift aufbewahrt.118 Aus dem letzten Drittel des 15. Jahrhunderts stammt schließlich auch ein umfangreiches deutschsprachiges Evangelistar (Innsbruck, ULB Tirol, Cod. 765, dat. 1472), aus dem 16. Jahrhundert hat sich eine volkssprachliche Fronleichnamspredigt (Innsbruck, ULB Tirol, Cod. 47) erhalten. 2.8 Provenienzen und Besitzeinträge Bei vielen Neustifter Handschriften lässt sich nicht genau feststellen, wann und auf welchem Weg sie Eingang in die Stiftsbibliothek gefunden haben. Nur zwei Codices weisen einen expliziten Schen kungsvermerk auf119; für eine Handschrift (Cod. 4) ist eine testamentarische Überlassung nach dem Tod des einstigen Besitzers festgeschrieben. Für weitere vier Handschriften konnte eine Schenkung dank einer erhaltenen Urkunde im Stiftsarchiv nachgewiesen werden. Die ersten handschriftlichen Besitzeinträge von Seiten des Stiftes reichen ins 15. Jahrhundert zu rück: Elf Handschriften weisen entsprechende Hinweise auf, teilweise sogar in zweifacher Ausführung; ihr Wortlaut ist stets ähnlich: Iste liber est monasterii sancte Marie vulgo in Novacella.120 Dem Schrift vergleich nach kann vermutet werden, dass sie wohl von einer einzigen Hand eingetragen wurden, 117 118 119 120 Der Entstehungsort der Handschrift ist umstritten; eine Entstehung in Thüringen (Schmalkalden) ist wahrscheinlich, vgl. Katalog ULB Tirol X (wie Anm. 100) 64–68 mit ausführlicher Bibliografie. Dieselbe Notiz findet sich auch in der sogenannten Sterzinger Miszellaneen-Handschrift, die heute im Stadtarchiv Sterzing aufbewahrt wird und deren Entstehungsort ebenso nicht eindeutig geklärt ist. Cod. 123, Bl. Vv und Cod. 748, Bl. IIIv. Mit dieser Schenkung dürfte vermutlich auch die Übergabe von Cod. 453 einhergegangen sein. Cod. 52, HDS; Cod. 748, Bl. 219r. Andere Besitzeinträge lauten: Iste liber est monasterii sancte Marie vulgo ad Novamcellam (Cod. 52, Bl. 2r; Cod. 107, Bl. 1r; Cod. 155, Bl. 164v; Cod. 485, Bl. 66v); Iste liber est monasterii sancte Marie ad Novamcellam (Cod. 78, Bl. 119v); Iste liber est monasterii sancte Marie ad Novamcellam prope Brixinam (Cod. 83, Bl. 56r); Iste liber est monasterii sancte Marie ad gracias alias ad Novamcellam dicti (Cod. 158, Bl. IIr); Iste liber est monasterii sancte Marie virginis ad gracias alias ad Novamcellam dicti (Cod. 617, Bl. Ir); Iste liber est monasterii sancte Marie in Novacella (Cod. 617, Bl. 281v); Iste liber est monasterii sancte Marie vulgo ad Novamcellam Brixinensis dyocesis (Cod. 453, VDS); Iste liber est monasterii sancte Marie in Novacella extra civitatem Brixinensem sita (Cod. 200, Bl. 377rb). Übersicht über den Handschriftenbestand 35 möglicherweise von Johannes Librarius (gest. 1467)121, dessen Beiname auf eine bibliothekarische Tätigkeit schließen lässt. Weitere handschriftliche Einträge reichen ins späte 17. bzw. frühe 18. Jahrhundert, als die Zuge hörigkeit ad Collegium Neocell. betont wurde. Insgesamt 13 Handschriften weisen einen entsprechen den Eintrag auf.122 Auch hier fällt der durchwegs ähnliche Wortlaut auf; mit Ausnahme von zwei Hand schriften123 dürften die Einträge von einer Hand gemacht worden sein. Wohl im späten 18. Jahrhundert und damit nach der Neugestaltung des Bibliothekssaals wurde in fast allen Neustifter Handschriften am Spiegel des Vorderdeckels ein Bucheignerzeichen eingeklebt: Der Großteil der Exlibris weist einen Zierrahmen auf und führt im Genitivus possessivus die Bibliothecae Novacellensis als Besitzerin an, darunter ist ein schlichtes Tau-Kreuz zu erkennen, das Wappenzeichen des Stiftes. Dieses Exlibris in Holzschnitttechnik, das sich auch in zahlreichen Drucken nachweisen lässt, ist ebenso wenig wie das zweite nachweisbare Bucheignerzeichen in Holzschnitt technik datiert. Letzteres zeichnet sich durch eine schlichte Rahmung aus, der Text sowie das Wappen zeichen sind identisch, und dürfte wohl ein paar Jahrzehnte später in Verwendung gekommen sein. Einige wenige Handschriften weisen einen zusätzlichen Besitzvermerk in Form eines Neustifter Rundstempels am Beginn der Handschrift auf. In jenen Codices, die nach Innsbruck gebracht worden waren, wurde zudem meist ein Rundstempel der dortigen Universitätsbibliothek angebracht; bei der Rückkehr nach Italien drückte auch die „Soprintendenza alle belle arti per la Venezia Tridentina“ ihren Stempel hinein. Leider wurden die Stempel nicht immer mit ausreichender Sorgfalt angebracht, sodass sie vereinzelt auch Texte abdecken. 121 122 123 Vgl. Anm. 51. Collegii Neocell. (Cod. 53, Bl. 1r; Cod. 158, Bl. IIIr; Cod. 177, Bl. 1r; Cod. 405, Bl. Iar; Cod. 440, Bl. 1r; Cod. 453, Bl. 1r; Cod. 550, Bl. 1r); Ad Collegium Neocell. (Cod. 52, Bl. 2r; Cod. 204, Bl. 1r; Cod. 253, Bl. 1r); Collegii Novacellensis (Cod. 4, Bl. 1r; Cod. 397, Bl. 1r); Ad Bibliothecam Neocell. (Cod. 748, Bl. 1r). Die Einträge in Cod. 4 und 397 lassen eine zweite Hand vermuten. 37 3. Besonderheiten der Erschließung und ihre Ergebnisse 3.1 Wasserzeichen Maria Stieglecker Für die Erfassung der Wasserzeichen in den Papierhandschriften der Stiftsbibliothek Neustift wurde die Vorgehensweise gewählt, die seit den 1970er Jahren an der ÖAW entwickelt wurde und sich bereits bei mehreren Handschriftenerschließungsprojekten bewährt hat.124 Ziel einer solch genauen Analyse ist, den wahrscheinlichen Beschreibzeitraum des Papiers zu ermitteln, Hinweise auf den Beschreibort zu finden und mögliche Beziehungen zwischen einzelnen Handschriften oder Handschriftengruppen aufzuzeigen. Um aussagekräftige Ergebnisse zu erzielen, werden sämtliche Blätter einer Handschrift auf ihre Wasserzeichen hin durchgesehen und ihre Standorte festgehalten, d. h. die Verteilung der verwendeten Papiersorten festgestellt. Diese genaue Durchsicht erweist sich als notwendig, da immer wieder einzel ne Blätter, etwa bei einer Neubindung, nachträglich eingefügt oder älteres Restpapier, das über Jahre im Skriptorium liegengeblieben war, in Einzelbögen oder ganzen Lagen untergemischt worden ist. Wird eine nur stichprobenartige Durchsicht nach Wasserzeichen vorgenommen, kann solches Papier zu einer Verfälschung der Datierung führen.125 Weiters kann über die Verteilungsübersicht des verwende ten Papiers die Zusammensetzung einer Handschrift verifiziert werden, da ein Papierwechsel häufig den Beginn einer neuen kodikologischen Einheit markiert.126 Von jeder Gruppe identischer Wasserzei chen eines Manuskripts bzw. eines seiner Teile wurde je nach den gegebenen Möglichkeiten eine Beta radiografie-Aufnahme, eine Abreibung oder ein Digitalfoto angefertigt. Diese Abbildungen der Papier marken und die zu den Trägerhandschriften erhobenen Daten wie Provenienz, Datierung oder Lokalisierung wurden in die Datenbank „WZMA – Wasserzeichen des Mittelalters“127 eingepflegt, wodurch eine gegenseitige Inbezugsetzung aller in WZMA erfassten Papiermarken ermöglicht wird. Die digitale Aufbereitung der Bilder und Daten erlaubt die dynamische Verknüpfung verwandter 124 125 126 127 A. Haidinger, Datieren mittelalterlicher Handschriften mittels ihrer Wasserzeichen. Anzeiger der phil.-hist. Klasse der ÖAW 139 (2004) 5–27. Als Beispiel sei Cod. 155 genannt, eine Sammelhandschrift, deren Entstehungszeitraum mit „um 1463“ einzuordnen ist, wie auch der Wasserzeichenbefund bestätigt. Nur wenige Blätter fallen aus der Reihe: Im Septernio Bl. 51–64 zeigen die inneren fünf Doppelblätter Wasserzeichen, die auf einen Beschriftungszeitraum um 1425/1435 schließen lassen. Hier kam allerdings kein Restpapier zum Einsatz, sondern es wurden wohl einige Blätter aus einer früher angelegten Handschrift eingebunden. Untermauert wird dies durch die Feststellung, dass der betroffene Text (Bl. 51ra–105r) bis auf die genannten Doppelblätter von einer Hand geschrieben wurde. Cod. 52 zum Beispiel besteht aus zwei Teilen, wobei Teil II mit 1396 datiert ist und Teil I nach Wasserzeichenbefund mit Ende 14. Jahrhunderts eingeordnet wird. Dieser Teil weist ein Wasserzeichenpaar Ochsenkopf mit Blume auf, während in Teil II ein dazu unterschiedliches Paar Ochsenkopf mit Blume zu finden ist sowie ein Paar Ochsenkopf ohne Beizeichen und zwei Paare Glocken. www.wzma.at 38 Besonderheiten der Erschließung und ihre Ergebnisse asserzeichen, sodass auf jeweils aktuellem Stand ersichtlich ist, für welche Handschriften Papier aus W derselben Produktionscharge verwendet wurde.128 Für verlässliche Untersuchungen von Wasserzeichen im Hinblick auf Aussagen zum Verwendungs zeitraum bestimmter Papiere ist eine repräsentative Menge an Vergleichsmaterial vonnöten. Erst aus mehreren unterschiedlichen datierten Quellen vorliegende identische Wasserzeichen erlauben eine sichere zeitliche Einordnung undatierten Materials.129 Umfangreiche Untersuchungen zu Wasserzeichen in Papierhandschriften, die sich zum einen in Sammlungen in Nord- und Südtirol befinden, zum anderen in Bibliotheken und Archiven in Wien und Niederösterreich, zeigen, dass in den jeweiligen Regionen unterschiedliches Importpapier aus Italien gehandelt und verwendet wurde. Eine Beobachtung, die durch die Analyse des Neustifter Bestandes bestätigt wird. Weiteres Material steuert hier die Sammlung Piccard130 bei, die Tausende von Wasser zeichen aus Tiroler Archivalien nachweist.131 Gerade die Auswertung von Wasserzeichen in Archiv alien aus dem Bereich der Verwaltung und Wirtschaftsaufzeichnungen tragen maßgeblich dazu bei, Fragen zu Datierung und Lokalisierung zu klären. Ihrem Verwendungszweck nach weisen sie in der Regel Datierungen auf und liefern über ihre lokale Verwendung und Aufbewahrung Hinweise auf Beschreiborte. Der dabei gesammelte bzw. über die Sammlung Piccard zur Verfügung stehende große Pool an Vergleichsmaterial erbrachte erwartungsgemäß viele Treffer für die Papiermarken der Neustifter Hand schriften. Sehr genau konnte z. B. der wahrscheinliche Beschreibzeitraum für Cod. 78 und 177 nach gewiesen werden, bei denen das verwendete Papier zwei Paare des Motivs Mohrenkopf mit Varianten zeigt. Allein in der Sammlung Piccard ergeben sich für diese Marken 56 Treffer, die folgende Datierun gen aufweisen: 1 × 1427, 7 × 1429, 18 × 1430, 11 × 1431, 8 × 1432, 4 × 1433 und 1 × 1435. In der Biblio thek des Stiftes Wilten findet sich ebenfalls eine Handschrift, deren Papier beide Paare zeigt: der mit der Datierung 1431 versehene Cod. 32 02 05 (Teil IV). Jeweils eines der genannten Wasserzeichen paare findet sich in einer ganzen Reihe weiterer Handschriften verschiedener Sammlungen, so in den 128 129 130 131 Die Sammlung WZMA enthält nach derzeitigem Stand knapp 19.000 Wasserzeichenbelege, haupt sächlich aus Handschriften und Archivalien, die sich in Bibliotheken und Sammlungen im Raum Wien / Niederösterreich sowie im geografischen Raum Tirol (Nord- und Südtirol) befinden. Im Zuge von Handschriftenerschließungsprojekten erfolgen laufend Ergänzungen. Eine Liste von Sammlungen, für deren Bestände bereits Wasserzeichen erhoben wurden, bietet der Reiter „Sammlungen“ auf www.wzma.at. Als Partner in das EU-Projekt „Bernstein“ (eContentPlus, 2006–2009) eingebunden, kann die Sammlung auch über das Portal „Bernstein – The Memory of Paper“ (www.memoryofpaper.eu) angesteuert werden. Nach Gerhard Piccard wie Theodor Gerardy wäre von einem Beschriftungszeitraum des Papiers innerhalb von vier Jahren ab Produktion auszugehen. Statistische Auswertungen für die Sammlung WZMA haben allerdings ergeben, dass Papier mit identischen Wasserzeichen durchaus auch über einen Zeitraum von sechs bis zehn Jahren – weniger häufig auch über eine noch größere Zeitspanne – Verwendung fand. Je mehr Vergleichsmaterial aber vorliegt, umso eher können „Ausreißer“ erkannt und für eine Datierung vernachlässigt werden; d. h. es kann ein Zeitraum festgestellt werden, in dem das Papier vorwiegend beschrieben worden ist; vgl. Haidinger, Datieren mittelalterlicher Handschriften 17–20. Die von Gerhard Piccard (1909–1989) angelegte Wasserzeichensammlung umfasst etwa 92.000 Karteikarten und wird am Hauptstaatsarchiv Stuttgart (Bestand J 340, Wasserzeichensammlung Piccard) verwahrt. Knapp zwei Drittel der Sammlung wurde über die 17 „Findbücher“ (1961–1997) in gedruckter Form zugänglich gemacht, in den Jahren 2003–2006 wurde die Sammlung digital aufbereitet und als Onlinefindmittel zur Verfügung gestellt, mittlerweile ist dieser Piccard-Online in das „WZIS – Wasserzeichen-Informationssystem“ (https://www.wasserzeichen-online.de/wzis/struktur.php) integriert; vgl. etwa P. Rückert, E. Frauenknecht (Hrsg.), Wasserzeichen und Filigranologie. Beiträge einer Tagung zum 100. Geburtstag von Gerhard Piccard (1909–1908). Stuttgart 2011. G. Piccard, Piccard Wasserzeichen. Veröffentlichungen der Staatlichen Archivverwaltung Baden-Württemberg. Sonderreihe: Die Wasserzeichenkartei Piccard im Hauptstaatsarchiv Stuttgart. 17 Findbücher in 25 Bänden. Stuttgart 1961–1997; Piccard: Hauptstaatsarchiv Stuttgart, Bestand J 340, Wasserzeichensammlung Piccard. Online zugänglich unter https://www.wasserzeichen-online.de/wzis/struktur.php; M. Stieglecker, Wasserzeichen des Mittelalters (WZMA) und Piccard-Online. Vom gegenseitigen Nutzen, in: Wasserzeichen und Filigranologie (wie Anm. 130) 79–89, hier 80–81. Besonderheiten der Erschließung und ihre Ergebnisse 39 Bibliotheken des Priesterseminars Brixen in Cod. A.1 (dat. 1432/1433) und B.16 (4. Jz. 15. Jh.) sowie in der ULB Tirol in Innsbruck in Cod. 228 (Vorsatzbl., um 1430/1432), Cod. 234 (Teil VI, um 1430/1431), Cod. 236 (Teil I, dat. 1429), Cod. 437 (Teil II, dat. 1432), Cod. 451 (dat. 1431), Cod. 481 (dat. 1431), Cod. 522 (Teil IV, um 1430/1432), Cod. 548 (dat. 1424132), Cod. 666 (Teil III, dat. 1433) und Cod. 763 (Teil VII, um 1429/1433). Diese Handschriften aus Sammlungen in Nord- und Südtirol weisen zum Teil Lokalisierungen auf, die sämtlich auf den Raum Tirol weisen: 4 × Schnals, je 1 × Neu stift (?), Hall, Salern und Stams. Auch ein Drittel der oben genannten Verweise in der Sammlung Piccard zeigt Beschreiborte im Raum Tirol auf. In einigen Beständen des Raumes Wien/Niederösterreich sowie Salzburg finden sich ebenfalls Handschriften mit verwandten Wasserzeichen, deren Beschreiborte allerdings unbekannt bzw. mit Fragezeichen versehen sind: Klosterneuburg, Stiftsbibliothek, CCl 300 (dat. 1431) und Cod. 683 (Teil V, um 1430), Wien, Schottenstift, Archiv HS 28 (Teil V, 4./5. Jz. 15. Jh.) und Archiv HS 377 (Teil I, dat. 1431), Wien, ÖNB, Cod. 5319 (dat. 1431–1432) und Michaelbeuern, Stiftsbibliothek, Man. cart. 65 (2. Viertel 15. Jh.). Dieses umfangreiche Vergleichsmaterial lässt den Schluss zu, dass die Neustifter Handschriften Cod. 78 und 177 nach Wasserzeichenbefund mit „um 1431/1433“ einzuordnen sind. Über Papier und Wasserzeichen der Neustifter Handschriften können Beziehungen des Stiftes zu Klosterneuburg bei Wien deutlich werden. Durch die bereits erfolgte Erfassung der Wasserzeichen mittelalterlicher Papierhandschriften etwa des Stiftes Klosterneuburg oder des Schottenstiftes in Wien ist hier gleichfalls eine breite Grundlage zur Recherche geboten. Als Beispiel sei der Neustifter Cod. 53 genannt: Papier mit Wasserzeichen der Motive Dreiberg mit Kreuz, Waage und Waage im Kreis, wie es für diese Handschrift verwendet wurde, findet sich ebenso in verschiedenen Handschriften bzw. Hand schriftenteilen und Archivalien des Stiftes Klosterneuburg sowie weiterer Sammlungen im Raum Wien/ Niederösterreich. Allein in der Stiftsbibliothek Klosterneuburg ist Papier mit diesen Marken in 20 Handschriften nachzuweisen, im Stiftsarchiv in zwei Grundbüchern und in zwei Rechnungs büchern. Diese Archivalien sowie eine der Handschriften sind eindeutig nach Klosterneuburg lokalisiert, bei zwei weiteren ist Wien als Entstehungsort im Kolophon genannt. In 20 weiteren Handschriften bzw. Handschriftenteilen der Sammlungen Schottenstift Wien, ÖNB, Haus-, Hof- und Staatsarchiv Wien, Benediktinerstift Melk und ULB Tirol wird als eindeutige Lokalisierung je einmal Wien und Gaming angegeben, je dreimal Wien sowie Niederösterreich als wahrscheinlicher Beschreibort. Damit kann für Cod. 53 ebenfalls eine Entstehung im Wiener Raum angenommen werden.133 Beziehungen zwischen Handschriften können sich auch in der Materialität widerspiegeln, wie die beiden Neustifter Manuskripte Cod. 453 und 748 zeigen. Bei beiden handelt es sich um medizinische Sammelhandschriften, die als Geschenk des Präbendars und Klosterarztes Bertoldus de Saxonia in den Besitz des Stiftes gelangten. Ein und derselbe Schreiber (möglicherweise Bertoldus de Saxonia selbst) ist über weite Strecken sowohl in Cod. 453 wie in Cod. 748 nachzuweisen: In Cod. 748 ist er als Haupt schreiber für die Teile I–III zu nennen, in Cod. 453 für die Teile II–IV. Für kleinere Texte, Notizen, Ergänzungen und Zählungen findet sich ein weiterer Schreiber sowohl für Teil I von Cod. 748 wie für Teil I von Cod. 453. Über Besitzer, Inhalt und Schreiber zeigt sich also eine Verschränkung der beiden Handschriften, gleichzeitig aber auch über das verwendete Material. Papier mit identischen Wasser zeichen des Motivs Dreiberg mit Kreuz wurde für die Teile I und III des Cod. 748 und Teil IV des Cod. 453 herangezogen. Ein Wasserzeichenpaar Horn findet sich in identischer Form in Cod. 748, Teil II und Cod. 453, Teil III. Jeweils in Teil III beider Handschriften zeigt das Papier ein weiteres Zeichen paar Dreiberg mit Kreuz, in Teil III von Cod. 748 sowie in Teil II von Cod. 453 ein Paar Horn. 132 133 Hier handelt es sich um ein nachträglich eingefügtes Einzelblatt, das besagtes Mohrenkopf-Wasserzeichen aufweist. Die Datierung dieses Codex kann damit für die Analyse vernachlässigt werden. M. Stieglecker, Verborgene Zeichen. Handschriftenbeschreibung und Wasserzeichenanalyse, in: B. Sára (Hrsg.), Quelle & Deutung I. Beiträge der paläographischen Tagung Quelle und Deutung I am 27. November 2013. Budapest 2014, 29–52, hier 46–48 (auch online verfügbar unter honlap.eotvos.elte.hu/wp-content/ uploads/2016/02/quelle.pdf). 40 Besonderheiten der Erschließung und ihre Ergebnisse Insgesamt wurden 50 Papierhandschriften des Bestandes Neustift auf ihre Wasserzeichen hin u ntersucht und 383 Abbildungen von Papiermarken in die Wasserzeichen-Datenbank WZMA ein gebunden. Mit diesen konnten für 47 bisher nur vage datierte Handschriften bzw. Handschriftenteile des Bestandes wesentliche Hinweise auf deren wahrscheinlichen Beschriftungszeitraum gegeben werden, weiters wurden die Datierungen der datierten Manuskripte über den Wasserzeichenbefund be stätigt. Auf der anderen Seite steht mit diesen Eingaben in WZMA für filigranologische Forschungen wieder mehr Vergleichsmaterial zur Verfügung, sodass zur Entstehungsgeschichte von Manuskripten im Tiroler Raum künftig immer genauere Ergebnisse zu erwarten sind. Systematische Analysen zu weiteren Beständen in der Region, seien es Handschriften, Inkunabeln oder Archivalien, böten hier Möglichkeiten, neben Fragen zu Datierung und Lokalisierung undatierter Papiere auch fundierte Aus sagen auf papiergeschichtliche Fragestellungen etwa im Hinblick auf Handelswege und die Verbreitung bestimmter Papiersorten134 zu treffen. 3.2 Illuminierte Handschriften Susanne Rischpler Der Bestand der Neustifter illuminierten Handschriften ist erstklassig, einige Stücke (siehe unten) sind als kostbare Zimelien einzustufen. Gut ein Viertel der Handschriften beinhaltet höherrangigen Buch schmuck. Daher wurde der Buchschmuckerschließung135 im vorliegenden Katalog gebührend Augen merk und Raum gewidmet – ohne jedoch den einfacheren Dekor zu vernachlässigen. Innerhalb der Katalogisate wird die Buchzier jeweils im Abschnitt A (Ausstattung) behandelt. Besteht eine Hand schrift aus mehreren Teilen, so wird im gegebenen Fall der Buchschmuck eines jeden Teils gesondert betrachtet. Die Erschließung orientiert sich an der Methode der „Mitteleuropäischen Schulen“ in der Katalogreihe zu den „Illuminierten Handschriften und Inkunabeln der Österreichischen National bibliothek“.136 Die Erfassung des Buchschmucks erfolgt nach hierarchischen Gesichtspunkten: Zuerst 134 135 136 Vgl. hierzu etwa M. Zaar-Görgens, Champagne – Bar – Lothringen. Papierproduktion und Papierabsatz vom 14. bis zum Ende des 16. Jahrhunderts. Trier 2004; R. Graziaplena, Paper Trade and Diffusion in Late Medieval Europe. A First Approach, in: R. Graziaplena (Hrsg.), Paper as a Medium of Cultural Heritage. Archaeology and Conservation (26th IPH-Congress. Rome, Verona, August 30th–September 6th 2002). Roma 2004, 343–354; M. Stieglecker, Papier in Tirol und Niederösterreich, in: Papers of the XXXII International Congress of Paper Historians: Fabriano – Amalfi 21–27 September 2014. Waimes 2018, 233–239. Zu Einsatzmöglichkeiten der Wasserzeichenforschung, zu Papiergeschichte oder Aufnahmeverfahren vgl. Ochsenkopf und Meerjungfrau. Papiergeschichte und Wasserzeichen vom Mittelalter bis zur Neuzeit. Begleitbuch und Katalog zur Ausstellung des Landesarchives Baden-Württemberg, Hauptstaatsarchiv Stuttgart und der Österreichischen Akademie der Wissenschaften, Kommission für Schrift- und Buchwesen des Mittelalters, Wien. Stuttgart und Wien 32009 (auch als PDF auf www.wzma.at unter „Materialien“ einzusehen). Ohne die umstandslose Unterstützung und den wertvollen fachlichen Rat der Kollegenschaft wäre die Buchschmuckerschließung nicht zu leisten gewesen. Mein herzlicher Dank gilt daher Ursula Stampfer und Claudia Schretter-Picker sowie Ulrike Bauer-Eberhardt (München, BSB), Regina Cermann (Wien, ÖAW), Giulia Gabrielli (Freie Universität Bozen), Katharina Hranitzky (Wien, Kunsthistorisches Institut), Edith Madas (Budapest, Országos Széchényi Könyvtár), Walter Neuhauser (gest. 2016), Karl-Georg Pfändtner (Augsburg, Staats- und Stadtbibliothek), Martin Roland (Wien, ÖAW) und Michaela Schuller-Juckes (Wien, Kunsthistorisches Institut). Nicht zuletzt danke ich der Bibliotheksstiftung Otto Pächt, Wien, für den Zugang zu ihrem reichen Bilderfundus. Bislang erschienen, jeweils in Wien (als Bände 10–14 der Reihe „Die Illuminierten Handschriften und Inkunabeln der Österreichischen Nationalbibliothek“): Mitteleuropäische Schulen I (ca. 1250–1350), 1997; Mitteleuropäische Schulen II (ca. 1350–1410): Österreich, Deutschland, Schweiz, 2002; Mitteleuropäische Schulen III (ca. 1350–1400): Böhmen, Mähren, Schlesien, Ungarn (mit Ausnahme der Hofwerkstätten Wenzels IV. und deren Umkreis), 2004; Mitteleuropäische Schulen IV (ca. 1380–1400). Hofwerkstätten König Wenzels IV. und deren Umkreis, 2014; Mitteleuropäische Schulen V (ca. 1410–1450): Wien und Niederösterreich, 2012. Besonderheiten der Erschließung und ihre Ergebnisse 41 werden die übliche Ausstattung und der niederrangige Dekor, wie Überschriften, Paragraphzeichen, Zeilenfüllsel, Zeigehände und Lombarden summarisch angeführt, dann wird, soweit vorhanden, der höherrangige Dekor ausführlich beschrieben, wobei sowohl für den ornamentalen als auch für den figürlichen Schmuck die stilistischen Charakteristika herausgearbeitet werden. Darüber hinaus wird gegebenenfalls auf stilistische und ikonografische Zusammenhänge (Vergleichsbeispiele) hingewiesen. Wenn sich aus der stilistischen und ikonografischen Einordnung stichhaltige datierungs‑ und/oder lokalisierungsrelevante Fingerzeige ergeben, fließen diese in den Katalogisatkopf der jeweiligen Be schreibung ein. Lediglich eine der insgesamt 92 in den Katalog aufgenommenen Handschriften (Cod. 578) trägt keinen nennenswerten Buchschmuck. 66 Codices weisen niederrangigen Buchschmuck auf. Zu diesem Ausstattungsniveau gehören neben der Rubrizierung, zu der von Überschriften bis hin zu Paragraph zeichen alle farblichen Hervorhebungen zählen, beispielsweise Zeilenschlussfüllungen, aufwändigere Zeigehände sowie verzierte Cadellen und Lombarden. Auch diese werden sorgsam erfasst, da sie immer wieder zur Gruppierung und Identifizierung von Schreibern, Rubrikatoren, Lombardenmalern und auch Illuminatoren beitragen können. 25 Handschriften sind mit höherrangigem Buchschmuck verziert, der von sorgfältig ausgeführten Fleuronnée-Lombarden und ‑Initialen, über kolorierte Federzeichnungen, ornamentale und figürliche Deckfarbeninitialen bis hin zu ganzseitigen Miniaturen reicht. Da sie sehr qualitätvoll bzw. ausgespro chenreich mit ornamentalem wie mit figürlichem Schmuck ausgestattet sind, werden folgende Codices ausführlicher behandelt: das Missale des Propstes Hieronymus Posch (Cod. 100), das „Speculum humanae salvationis“ (Cod. 166), das Rituale Cod. 194, das Lektionar Cod. 196, das Ordinarium missae Cod. 252, das zweibändige Stundenbuch des Klausner Stadtrichters Kaspar Neuhauser (Cod. 353 und 654), das Brevier des Propstes Lukas Härber von Ringelsperg (Cod. 405), das Rituale Sign. 14820 sowie die beiden Bände des berühmten Neustifter Graduales (ohne Sign.). Die meisten dieser Handschriften wurden im Jahr 1905 von Hermann Julius Hermann erstmals unter kunsthistor ischen Gesichtspunkten erfasst.137 Ein gutes Viertel und damit die meisten der in der Neustifter Bibliothek aufbewahrten illuminierten Handschriften stammen wohl aus Neustift selbst oder aus Brixen, das Gros davon entstand im 15. und 16. Jahrhundert. So sind bis auf das „Speculum humanae salvationis“, das zu Beginn des 15. Jahr hunderts im süddeutschen Raum geschaffen worden sein dürfte, auch alle Zimelien mit Sicherheit oder größerer Wahrscheinlichkeit nach Neustift zu lokalisieren, wo sie an der Wende vom Spätmittelalter zur Frühen Neuzeit geschrieben und ausgeschmückt wurden. Durch die Untersuchung des Buchschmucks konnten in nicht wenigen Fällen die Datierung und auch die Lokalisierung der Handschriften präzisiert werden: Die Miszellanhandschrift Cod. 76 enthält zwar nur eine größere Initiale, doch trägt diese dazu dabei, den entsprechenden Handschriftenteil nach Neustift zu lokalisieren und in die Mitte des 15. Jahrhunderts zu datieren, da die Initiale von der Hand des Hauptflorators der Neustifter Gradualien stammt. Mit Hilfe des Fleuronnées lassen sich Hand schriften zudem gruppieren, wodurch wiederum Datierungen ermöglicht werden. So haben u. a. die teilweise datierten Neustifter Liturgica Cod. 405 und 194 sowie die Sammelhandschrift Cod. 142 (Datierungen von 1501 bis 1521) eine Floratorenhand gemeinsam, deren Hauptwerk, das bis auf spätere Zusätze undatierte Rituale Sign. 14820, somit in das erste Fünftel des 16. Jahrhunderts gegeben werden kann. 137 H. J. Hermann, Die illuminierten Handschriften in Tirol (Beschreibendes Verzeichnis der illuminierten Handschriften in Österreich 1). Leipzig 1905. Hermann beschreibt ausführlich die damals noch in der Universitätsbibliothek Innsbruck befindlichen Cod. 4, 76, 99, 100, 166, 194, 196, 217, 252 und 654 sowie die beiden in Neustift verbliebenen Gradualbände (die genauen bibliografischen Verweise zu Hermann siehe im vorliegenden Katalog jeweils im Literaturabschnitt der Beschreibungen). 42 Besonderheiten der Erschließung und ihre Ergebnisse Um die Mitte des 15. Jahrhunderts wird das buchmalerische Geschehen durch das Großprojekt der beiden Gradualbände beherrscht, die sehr wahrscheinlich vor Ort in Neustift entstehen. Das Vorhaben bindet die Mitarbeiter aller Ausstattungsebenen für längere Zeit, sie können ihre Fähigkeiten entfalten und werden dann gerne für weitere illuminatorische Tätigkeiten herangezogen – sowohl in Neustift selbst als auch in Brixen. So ordern die Brixner Bischöfe der Zeit, Johannes Röttel (1444–1450) und wohl auch Nikolaus Cusanus (1450–1464), Liturgica, die sie von Mitarbeitern der Gradualbände aus statten lassen. Schöne Beispiele, die Zeugnis für die enge Verbindung von Neustift und Brixen in dieser Phase ablegen, sind die Missalien Cod. C.1 und F.8 der Brixner Priesterseminarbibliothek, in denen sich Illuminationen von der Hand des Servatius, eines der Deckfarbenmaler der Neustifter Gradualien, finden. Der von Bischof Röttel in Auftrag gegebene Cod. C.1 ist 1447 datiert, entstand also, nachdem bereits ein Großteil der Ausstattungsarbeiten an den Gradualbänden abgeschlossen gewesen sein dürfte. Des Weiteren schmückte auch der zweite Deckfarbenmaler der Neustifter Gradualien bischöfliche Li turgica aus, und zwar das Brevier Cod. E.21 und das Missale Cod. F.4 der Brixner Priesterseminarbib liothek, die keine expliziten Datierungen tragen, sich aber dank der Zusammenhänge im Buchschmuck kurz vor die Mitte des 15. Jahrhunderts datieren lassen. Diese bereits vor der aktuellen Katalogisierung bekannten Verbindungen auf der illuminatorischen Ebene machten deutlich, dass nur eine eng verzahn te Erschließung der Handschriftenbestände in Neustift und in der Brixner Priesterseminarbibliothek eine sinnvolle sein konnte: So zeigt sich nicht nur das 1480 datierte Brevier des Erasmus Pugsinger (Priester seminarbibliothek, Cod. T.3) im Deckfarbenschmuck, der naturalistische, goldverzierte Blütenzweige variiert, mit dem Lektionar Cod. 196 verwandt und stützt dessen Datierung, sondern auch das Brixner Graduale Cod. F.5, für das sich somit zumindest eine Datierung ans Ende des 15. Jahrhundert recht fertigen lässt. Auch die annähernd 3000 Initialen und Cadellen, die der Hauptflorator der Neustifter Gradualien mit seinem akkuraten Fleuronnée verzierte und mit unzähligen charakteristischen Profil köpfen besetzte, übten einen starken Einfluss aus, z. B. auf das Cadellen-Fleuronnée im Brixner Missale Cod. C.1 oder noch in der um einige Jahrzehnte jüngeren Neustifter Bruno de Segni-Handschrift Cod. 190. Berührungspunkte ergeben sich auch auf der Ebene der Lombarden. Im ersten Band des Neuhauser-Stundenbuchs (Cod. 353), das im Deckfarbendekor einige Affinitäten zur Neustifter Buch malerei zeigt, beweist der Lombardenschmuck Verbindungen zu den aufwändiger ausgeführten Lom barden in den Brixner Pontifikalien Cod. C.8 und Cod. F.6. Da Cod. 353 durch seinen 1496 datierten zweiten Band mit Sicherheit um oder kurz nach 1496 einzuordnen ist, kann so für die beiden Brixner Codices eine Datierung ans Ende des 15. Jahrhunderts ins Auge gefasst werden. Ein Ergebnis der kunsthistorischen Untersuchung des Neustifter Bestandes, das es hervorzuheben gilt, ist die Entdeckung des Servatius, bei dem es sich mit sehr großer Wahrscheinlichkeit um die erste Illuminatorenhand der Neustifter Gradualien handelt. Dieser Buchmaler führte auch Fleuronnée aus, ist aber nicht mit dem Hauptflorator der Gradualien gleichzusetzen, der wahrscheinlich gemeinsam mit Friedrich Zollner, dem Schreiber, Rubrikator und Notenmaler der Gradualien, nach Neustift kam. Eigentlich muss man im Fall des Servatius von einer Wiederentdeckung sprechen. Er war in Vergessen heit geraten, obwohl er seine mit einem Bildnis verknüpfte Signatur Servacius Lichtensteyn de Prussia im zweiten Gradualband hinterlassen hatte und in der Literatur bereits sporadisch als Illuminator genannt worden war. Während der Schreiber Friedrich Zollner, der in der Literatur immer wieder irrigerweise als Illuminator vereinnahmt wurde, nachweislich aus Langenzenn bei Nürnberg kam138, kann man von Servatius nur vermuten, auf welchen Pfaden er nach Neustift gelangte. Mutmaßlich war er ein wan dernder Buchmaler. Es ist folglich offensichtlich, dass man für das Gradualienprojekt leistungsfähige und begabte Buchkünstler – sowohl Kleriker wie den Augustiner Chorherrn Zollner als auch Laien wie 138 Zur Gebetsbrüderschaft zwischen den Augustiner Chorherrenstiften Neustift und Langenzenn vgl. Anm. 14 und 18. Besonderheiten der Erschließung und ihre Ergebnisse 43 Servatius – von außerhalb heranzog.139 Diese Vorgehensweise war nicht unüblich. Als beispielsweise in der ersten Hälfte der 1420er Jahre im niederösterreichischen Klosterneuburg ein vierbändiges Anti phonar in Angriff genommen wurde, holte man sich ebenfalls externe Kräfte, was in diesem Fall durch die Eintragungen in den Klosterneuburger Rechnungsbüchern gut belegt ist.140 Der umfangreiche Neustifter Handschriftenbestand des 15. und 16. Jahrhunderts birgt nicht zu letzt anschauliche Beispiele für den Umgang mit Vorlagen in einem Skriptorium dieser Zeit. Eine wichtige Rolle spielte dabei das bereits genannte Lektionar Cod. 196, welches sich aus einem Epistolar und einem unvollständigen Evangelistar zusammensetzt. Da es möglich war, den fehlenden Beginn des Evangelistarteils, der sich seit dem 19. Jahrhundert in der Kathedralbibliothek von Esztergom (Ungarn) befindet, in die Untersuchung einzubeziehen141, konnte klar herausgearbeitet werden, dass Epistolar und Evangelistar von je einem Buchmaler ausgeschmückt worden waren, die auf einen gemeinsamen Vorlagenfundus – eventuell ein Musterbuch – zurückgegriffen haben.142 Diese Vorlagen wurden in Neustift so gut gehütet, dass sie noch in der Renaissancezeit wirksam werden konnten. Daher inspirier te Cod. 196 nicht nur in den 1490er Jahren die Seitenranddekorationen in den zusammengehörigen Brevierbänden Cod. 353 und 654 (dat. 1496), sondern lieferte noch in den 1520er Jahren Modelle für das exquisite Posch-Missale – und zwar vom Randschmuck mit seinen naturalistisch anmutenden Pflanzendarstellungen bis hin zu den Kompositionen figürlicher Initialen. Selbstverständlich diente in dieser Zeit auch Gedrucktes als Grundlage für gemalten Buchschmuck. So wurden für den Großteil der Vollbilder im Brevierband Cod. 654 vermutlich die Illustrationen einer knapp zehn Jahre zuvor erschie nenen Inkunabel als Grundlagen benutzt, wobei man auch die dreiregistrige Unterteilung der Bildfelder übernahm.143 Von einer Inkunabel mit gemaltem Buchschmuck wurden wiederum Cod. 196 und der in Esztergom befindliche Handschriftenteil beeinflusst, und zwar in Bezug auf Maltechnik und Motiv repertoire. Der Schmuck dieser Inkunabelbibel dürfte seinerseits in der Nachfolge des Innicher Missales stehen, das sich im Verlauf der Buchschmuckerschließung in Neustift und Brixen als ein weiteres maß gebliches Werk für die Buchmalerei in Neustift-Brixen und Umgebung herauskristallisierte.144 Daher sind weitere Projekte zur Erschließung von Handschriftenbeständen sowie zur Erforschung von Inku nabelbeständen in (Süd-)Tiroler Bibliotheken nur zu begrüßen, da sie mit Sicherheit dazu beitragen werden, die Buchschmuckbezüge zwischen den erhaltenen Handschriften im Tiroler Raum zu erhellen. 139 140 141 142 143 144 Zu Servatius (und zu Zollner) siehe S. Rischpler, „Servacius Lichtensteyn de Prussia“, in: J. F. Hamburger, M. Theisen (Hrsg.), Unter Druck. Mitteleuropäische Buchmalerei im 15. Jahrhundert. Tagungsband zum internationalen Kolloquium in Wien, ÖAW, 13.–17.1.2016 (Buchmalerei des 15. Jahrhunderts in Mitteleuropa 15). Petersberg 2018, 41–52. Vgl. S. Rischpler, Der Illuminator Michael. Purkersdorf 2009 (Codices Manuscripti. Supplementum 1) 1 8–19, 46–49 (Nr. 1), 94. An dieser Stelle sei Edith Madas (Budapest, Országos Széchényi Könyvtár) nochmals für ihre freundliche Hilfe gedankt, die sowohl die Zusammensicht des Cod. 196 mit Esztergom, Kathedralbibliothek, Ms. II.2 als auch des Neustifter Cod. 146 mit Esztergom, Kathedralbibliothek, Ms. II.516 ermöglichte. Auch im Datierungskontext ist das Lektionar Cod. 196 von Interesse: In dem in Esztergom befindlichen Handschriftenteil begegnet, eingeschrieben in eine figürliche Deckfarbeninitiale, eine Jahreszahl, die für die Datierung der Gesamthandschrift richtungsweisend sein dürfte (Esztergom, Kathedralbibliothek, Ms. II.2, Bl. 5v: Darbringung im Tempel; an der Wand über den Gesetzestafeln die Jahreszahl 1482; die 2 abgerieben). Itinerarium seu peregrinatio BMV: lat., Johann Reger, Ulm, ca. 1487/88, GW M15415 oder dt., Johann Schobser, Augsburg, 1489, GW M15419 (jeweils mit Links zu Volldigitalisaten). Inkunabelbibel: Wien, ÖNB, Ink. 3.C.6; Johann Mentelin, Straßburg, vor dem 27. Juni 1466 bzw. um 1470, GW 4295 (http://data.onb.ac.at/rec/AC07662248, auf dieser Seite Link zum Volldigitalisat); zu dieser Bibel vgl. M. Roland, Buchmalerei der Gotik, in: P. Naredi-Rainer, L. Madersbacher (Hrsg.), Kunst in Tirol I (Kunstgeschichtliche Studien – Innsbruck N. F. 3). Innsbruck, Wien, Bozen 2007, 267–294, 291–292, Kat. 188. – Missale: Innichen, Kollegiatstift, ohne Sign., zu dieser Handschrift, die wohl in der 2. Hälfte der 1450er Jahre entstanden ist, siehe Hermann, Die illuminierten Handschriften (wie Anm. 137) 75–78, Nr. 81; E. Kühebacher, Kirche und Museum des Stiftes Innichen. Bozen 1993, Nr. 163, Abb. 116, 117a–g; Roland, Gotik 275 Anm. 79, Abb. 17. 44 Besonderheiten der Erschließung und ihre Ergebnisse 3.3 Musikalien Giulia Gabrielli Musik spielte in Neustift seit jeher eine große Rolle. Choralbücher, Musikarchivalien sowie die von 1390 bis in die Neuzeit nachweisbaren geistlichen Spiele dokumentieren den hohen Stellenwert der Musik im Kloster. Ins Jahr 1466 reicht die erste urkundliche Erwähnung eines Orgelspiels in der Stifts kirche zurück.145 Auch wenn zunächst wohl noch als curiositatis vanitas abgetan, bildet die Orgel seit her einen wichtigen Bestandteil der Kirche. Unter Propst Markus Hauser waren sogar vier Orgeln in der Kirche aufgestellt.146 Eine besondere Bedeutung kommt seit den Anfängen dem Choralgesang zu, der maßgeblich zur feierlichen Gestaltung der Liturgie beitrug. Die bis heute erhaltenen liturgischen Handschriften mit Notation dürften, wie nicht zuletzt auch die zahlreichen Fragmente belegen, wohl nur den Rest einer ursprünglich weit umfangreicheren Sammlung an Musikhandschriften bilden. Dennoch machen sie die große Musiktradition im Augustiner Chorherrenstift deutlich. „Es sind dort gereifte und ehrerbietige Männer, und ich glaube, ich hab nie einen genaueren und besseren Chorgesang als in diesem Kloster gehört.“147 Der Dominikaner Felix Faber betonte in seinem Reisetagebuch aus den Jahren 1483/84 nicht nur den reichen Bücherbestand von Neustift, sondern hob insbesondere auch die Qualität des liturgischen Gesangs hervor. Eine intensive musikalische Tätigkeit und die Produktion von Musikhandschriften zeichnen das Stift vermutlich schon seit den ersten Jahr hunderten seines Bestehens aus. Dies bezeugen u. a. Persönlichkeiten, die in Neustift lebten oder dort zu Besuch waren. Propst Albert (1298–1314) wird im „Memoriale benefactorum Novacellensium“ als metrista perfectus, musicus subtilis bezeichnet.148 Derselben Chronik zufolge hat Albert auch liturgi 145 146 147 148 A. und M. Reichling, Orgelgeschichte der Stiftskirche zu Neustift bei Brixen, in: Festschrift. Orgelweihe der Neuen Metzler-Orgel. Stiftskirche Neustift. Sonntag, 18. Mai 2014. Neustift 2014, 31–43, hier 31. Ebda 33. „Sunt ibi viri maturi et observantiales: numquam videtur mihi me audivisse tam regularem et bonum chori cantum, sicut in hoc monasterium.“ Vgl. Anm. 3. Johannes Librarius, Memoriale benefactorum Novacellensium, Hs. aus dem Jahr 1463, ursprünglich in Neustift verwahrt, heute verloren. Eine Transkription des auf Albert bezogenen Abschnitts ist enthalten in: A. Dörrer, Bozner Bürgerspiele. Alpendeutsche Prang- und Kranzfeste, Bd. 1. Leipzig 1941, 142. Vgl. auch M. Frei, Das Memoriale benefactorum des Augustiner-Chorherrenstiftes Neustift verfasst von Johannes Librarius, Diss. Wien 1960. Albert von Neustift wird für gewöhnlich die Urheberschaft der marianischen Se quenz Ave cella novae legis zugeschrieben, in deren erstem Vers angeblich die alte Bezeichnung des Klosters, „cella nova“, anklingt. In der Chronik des Johannes Librarius findet sich jedoch kein Hinweis auf bestimmte von Albert verfasste Werke, ebensowenig auf diese Sequenz. Eine Zuschreibung der marianischen Sequenz an Propst Albert erfolgte erstmals durch Anton Dörrer (vgl. Dörrer, Bozner Bürgerspiele 142). Er stützte sich dabei auf die Aussage des Kanonikers und Historikers Max Schrott, dass er diese Sequenz in einer Hand schrift des 14. Jahrhunderts aus Neustift gefunden und den Gleichklang zwischen der ersten Strophe und dem Namen des Stiftes bemerkt habe. Diese hypothetische Zuschreibung an Propst Albert wurde auch in der spä teren Literatur übernommen, vgl. u. a. A. Dörrer, Albert von Neustift (Albertus Novacellensis), in: Verfasser lexikon (wie Anm. 51) V, 21–22; L. Oberrauch, Musik, in: Tyrolis Latina (wie Anm. 51) 143–146, hier 144. Derzeit ist es nicht möglich, die von Max Schrott genannte Handschrift aus dem 14. Jahrhundert zu identifizieren. Die einzigen beiden im Augustiner Chorherrenstift Neustift gefundenen Zeugen für die Se quenz sind ein Fragment aus dem 14. Jahrhundert, das für den Einband von Cod. 440 verwendet wurde und am Rand zu einem Totenoffizium den Text der Sequenz enthält (siehe Beschreibung im Katalog), und die Ver sion mit Notation im zweiten Band des Zollner-Graduales aus dem 15. Jahrhundert (Bl. 217v). Jedenfalls aus zuschließen ist die von Oberrauch vorgeschlagene Übereinstimmung der von Schrott genannten Handschrift mit Cod. 710 der ULB Tirol, da diese höchstwahrscheinlich aus Wilten stammt und sich kein Bezug zu Neu stift herstellen lässt; vgl. Katalog ULB Tirol VIII (wie Anm. 100) 60–65; St. Engels, Einstimmige liturgische Handschriften des Mittelalters in Tirol, in: Musikgeschichte Tirols I (wie Anm. 43) 119–322, insbes. 293–296. Besonderheiten der Erschließung und ihre Ergebnisse 45 sche und musikalische Handschriften kompiliert.149 Im Hinblick auf die weltliche Musiktradition in Neustift darf die enge Beziehung des Minne sängers Oswald von Wolkenstein zu Neustift nicht unerwähnt bleiben. Möglicherweise ließ er im damals sehr renommierten Skriptorium des Stiftes die zwei Liederhandschriften herstellen, die sich heute in Wien und in Innsbruck befinden.150 In der ULB Tirol wird außerdem noch eine weitere berühmte Musikhandschrift des 15. Jahrhunderts aus Neustift verwahrt, nämlich Cod. 457. Die Handschrift über liefert eine umfangreiche Sammlung einfacher polyphoner liturgischer Musik; aufgrund der großen Bedeutung des enthaltenen Repertoires war die Handschrift in den vergangenen Jahrzehnten Gegen stand intensiver wissenschaftlicher Untersuchungen.151 Ein ganz anderes Schicksal erlebten die fünf mittelalterlichen Handschriften mit Kompositionen in Cantus planus, Cantus fractus und einfacher Polyphonie, die noch heute in der Stiftsbibliothek zu finden sind. Sie sind wichtige Zeugnisse für die Geschichte des liturgischen Gesangs in Neustift. Alle fünf Handschriften stammen aus dem 15. Jahrhundert – von den älteren Musikhandschriften ist keine vollständig erhalten geblieben; daneben existiert eine große Anzahl an Fragmenten, die erst vor Kurzem systematisch untersucht und beschrieben wurden.152 Die Musikhandschriften aus Neustift werden im vorliegenden Katalog zum ersten Mal vollständig und systematisch beschrieben.153 Drei davon – Nr. 139 (olim Sign. 14819) und die beiden Bände des Zollner-Graduales – waren in der Forschung bereits bekannt, während zwei – Nr. 940 und Sign. 15063 – erstmals genauer untersucht werden konnten. Alle fünf Handschriften enthalten musikalische Werke aus dem sogenannten Gregorianischen Repertorium in gotischer Choralnotation auf Vierliniensystem. Es handelt sich um zwei großformatige Choralbücher mit reichem Dekor, die gemeinsam das sogenannte Zollner-Graduale bilden und die über Jahrhunderte hinweg in der Stiftskirche verwendet wurden, sowie um drei kleinere und weniger aus geschmückte, von denen zwei womöglich für Kirchen in der Umgebung erstellt wurden. Die zwei großformatigen Bände des Graduale stellen wohl die berühmtesten in Neustift ver wahrten Handschriften dar. Es ist anzunehmen, dass die Chorsänger, von denen Felix Faber in den 1480er Jahren so begeistert war, aus ebendiesen, etwa 40 Jahre zuvor geschriebenen Handschriften gesungen haben. Wie bereits an anderer Stelle ausführlich dargelegt, entstanden die zwei Bände zwischen 1439 und 1446 und wurden vom Augustiner Chorherrn Friedrich Zollner aus dem Kloster Langenzenn geschrieben (siehe hierzu Abschnitt 3.2). In den reichen Buchschmuck, gestaltet von mehreren Händen, werden bisweilen auch Textzeilen und Notation eingebunden. So sind z. B. in der Initiale R von Resurrexit (Introitus zum Ostersonntag, Graduale Neocellense I, Bl. 156v) drei Schrift rollen mit den Initien der drei Ostergesänge dargestellt, einer davon zweistimmig.154 149 150 151 152 153 154 Claret in libris missalibus quibusdam […] et libris cantualibus („Memoriale benefactorum Novacellensium“, zit. nach Dörrer, Bozner Bürgerspiele [wie Anm. 148] 142). Es ist jedoch keine liturgisch-musikalische Handschrift aus der Zeit Alberts vollständig erhalten geblieben. Die erste (Cod. A) befindet sich in Wien, ÖNB (Cod. Vind. 2777), die zweite (Cod. B) in der ULB Tirol (Wolkenstein, o. S.). Vgl. Katalog ULB Tirol V (wie Anm. 100) 358–375, mit ausführlicher Bibliografie. Die Katalogisierung erfolgte im Rahmen eines von der Freien Universität Bozen finanzierten Forschungs projekts unter der Leitung von Giulia Gabrielli, vgl. Anm. 109. Aufgrund der erschwerten Zugänglichkeit berücksichtigte Praßl in seiner Arbeit über die Sequenzen der österreichischen Augustiner 1987 die Neustifter Musikhandschriften nicht (F. K. Prassl, Psallat ecclesia mater. Studien zu Repertoire und Verwendung von Sequenzen in der Liturgie österreichischer Augustinerchorherren vom 12. bis zum 16. Jahrhundert, Diss. Graz 1987). Jüngere Untersuchungen wie jene von Peintner, Schreibkunst (wie Anm. 43), insbes. 366–368, und von Engels, Einstimmige liturgische Handschriften (wie Anm. 148) behandeln nur Teile des vorhandenen Materials. Es handelt sich um den Hymnus Salve festa dies, die marianische Antiphon Regina celi und den zweistimmi gen Tropus Alle[uia] Dei filius. 46 Besonderheiten der Erschließung und ihre Ergebnisse Die detaillierte Erforschung des musikalischen Inhalts der beiden Handschriften steht noch am Anfang.155 Doch bereits bei einer ersten genauen Durchsicht fallen einige seltene oder gar einzigartige Kompositionen auf. Es handelt sich hierbei in erster Linie um Tropen, also Ergänzungen textlicher und/ oder melodischer Art, mit dem Ziel, Inhalt und Bedeutung der kanonisierten Stücke zu aktualisieren und weit verbreiteten Kompositionen lokale oder identitätsstiftende Merkmale zu geben. Gerade für das 15. Jahrhundert sind insbesondere im süddeutschen Raum eine Fülle solcher Tropen bekannt.156 Im Neustifter Graduale finden sich derartige Ergänzungen bei den Festen bzw. liturgischen Formularen für die wichtigsten Heiligen des Stiftes: für den Ordensvater Augustinus und für die Gottesmutter Maria. Die Initien eines Tropus zum Introitus Statuit ei beim Fest des hl. Augustinus (Graduale Neocellense II, Bl. 175r) sind in keiner anderen Quelle belegt (Initien: Augustinus* – Hic est manu* – Ut pro nobis* – Hic est cleri). Auch die Alleluia-Komposition in derselben Messe (Alleluia Protege pastor bone, Bl. 175r) hat großen Seltenheitswert, ihre Melodie ist auch im Graduale Magnum von ca. 1490 aus dem Augustiner Chorherrenstift Seckau überliefert.157 Die Beziehungen zwischen Neustift und Seckau reichen bis zur Gründung jenes Stifts zurück: Bischof Hartmann von Brixen, Gründer von Kloster Neu stift, nahm in seinem Todesjahr 1164 die Weihe der Stiftskirche von Seckau vor.158 Möglicherweise ist gerade das Vorhandensein derselben Komposition in beiden Augustiner Chorherrenstiften ein Zeichen für den engen Kontakt der Stifte auch noch im 15. Jahrhundert. Auch die Tropen zum Introitus Gau deamus für die Feste der Himmelfahrt Mariens (Graduale Neocellense II, Bl. 171v, Inc.: Quia virgo) und Mariä Geburt (Graduale Neocellense II, Bl. 186r, Inc.: Nata summa) stellen Unikate dar. Wie auch beim Fest des hl. Augustinus enthält die Handschrift nur die Textanfänge der Tropen, wohl ein Zeichen dafür, dass Text und Melodie bekannt waren und von den Sängern aus dem Gedächtnis vor getragen wurden. Ebenso einzigartig ist der kurze Tropus zum Agnus Dei O dulcis Iesu Christe mit rhythmischer Notation, der gleich in beiden Bänden vorkommt (Graduale Neocellense I, Bl. 305v und II, Bl. 288r). Weiters fällt die große Fülle an Sequenzen auf, die im Graduale überliefert werden – auch dies ein typisches Merkmal der deutschen Quellen des späten 15. Jahrhunderts. Die über 90 Sequenzen in beiden Bänden entsprechen im Wesentlichen den in zeitgenössischen österreichischen Augustiner Chorherren-Handschriften enthaltenen Sequenzen-Reihen und weisen signifikante Überschneidungen mit dem Repertorium von Seckau auf.159 Hervorzuheben sind zwei Sequenzen für die Brixner Diözesan patrone: Innoventur hac in die (Graduale Neocellense II, Bl. 107v) für das Fest der hl. Ingenuin und Albuin und Gloriosa fulget dies für jenes des hl. Kassian (Graduale Neocellense II, Bl. 274v–274br). Die erste Sequenz wurde eigens für diese Heiligen komponiert und wird auch in anderen Handschriften der Diözese Brixen überliefert160, im Repertorium anderer österreichischer Augustiner Chorherrenstifte ist sie hingegen nicht zu finden. Die zweite, Gloriosa fulget dies, taucht in ebendiesem Repertorium 155 156 157 158 159 160 Eine erste Untersuchung der Neustifter Tropen wurde von der Verfasserin für ihren Vortrag „Tropes in cantus planus sources from South Tyrol“, gehalten beim 20th International Musicological Society Congress, Study Group Cantus Planus, Venice, July 28th– August 1st 2014, durchgeführt. Der Beitrag wird im Tagungsband publiziert: J. Borders u. a. (Hrsg.), Cantus planus. Venice meeting 2014 (im Druck). Zur Verbreitung und zu den charakteristischen Merkmalen der Tropen aus jüngeren Quellen im deutschsprachigen Raum sowie über die verschiedenen Quellenarten siehe A. Haug, Troparia tardiva. Repertorium später Tropenquellen aus dem deutschsprachigen Raum (Monumenta monodica Medii Aevi. Subsidia 1). Kassel 1995, insbes. 3–10. Graz, UB, Hs. 17, Bl. 188v. Die Handschrift ist unter http://143.50.26.142/digbib/handschriften/Ms.00010199/Ms.0017/index12.html einsehbar. Vgl. A. Sparber, Leben und Wirken des seligen Hartmann, Bischofs von Brixen (1140–1164). Wien 1957, 82. Eine Verbrüderung des Augustiner Chorherrenstiftes Seckau mit Neustift ist seit 1331 belegt (vgl. Pallhuber, Der Neustifter Rotelbote [wie Anm. 18] 111). Diese und die beiden folgenden Beobachtungen basieren auf dem Vergleich mit Quellen und Repertorien aus Prassl, Psallat ecclesia mater (wie Anm. 153). G. Gabrielli, Katalog der in Bozen und Brixen verwahrten Musikhandschriften im Cantus planus und Cantus fractus, Diss. Graz 2015, 369, 483. Besonderheiten der Erschließung und ihre Ergebnisse 47 auf.161 Dort ist allerdings die Verwendung dieser üblicherweise für die Bekenner verwendeten Sequenz162 für das Fest der Translatio mehrerer Heiliger, insbesondere des hl. Rupertus, Virgilius, Nicolaus, Valentinus und Augustinus, belegt. Im Neustifter Graduale ist Gloriosa fulget dies gleich in zwei Versionen enthalten: für eben diese beiden Feste (Graduale Neocellense II, Bl. 272v–274v) und unmittelbar darauffolgend wie erwähnt für das Fest des hl. Kassian. Hier zeigt sie sich in einer wesent lich ausgefeilteren Form als in der vorangehenden Variante, reich mit Melismen verziert und mehreren Textvarianten; die auf den vorhergehenden Blättern angeführten Strophen 4 und 5 sind hier nun weg gelassen. Am Rand – vermutlich Ende des 18. Jahrhunderts – ergänzte Anmerkungen und Hinweise für die praktische Umsetzung lassen darauf schließen, dass das Graduale wohl über einen sehr langen Zeit raum in Neustift in Gebrauch war. An zahlreichen Stellen wurden die Melodien außerdem durch das Weglassen von Melismen verändert; diese wurden mit Papierstreifen überklebt oder mittels grafischer Zeichen wie senkrechte Striche in Rot oder Schwarz gestrichen. Die Herstellung von Choralbüchern wurde auch in neuzeitlicher Zeit in Neustift fortgesetzt, wie zahlreiche erhaltene Handschriften bele gen.163 Die beiden Papierhandschriften Nr. 139 (olim Sign. 14819) und Nr. 940 stammen von einer Hand und wurden Ende des 15. Jahrhunderts von Heinricus Edlinger geschrieben; beide enthalten ein Anti phonar sowie ein Graduale. Nr. 139 (olim Sign. 14819) wurde bereits von Stefan Engels genau erforscht:164 Die Handschrift enthält u. a. Beispiele für Cantus fractus in gotischer Choralnotation. Nr. 940 ist hingegen in der Forschung noch weitgehend unbekannt. Die Handschrift weist dieselben Merkmale und denselben Inhalt wie Nr. 139 (olim Sign. 14819) auf, möglicherweise diente sie als Vor lage für Nr. 139. Beide Handschriften dürften für zwei Kirchen in der (Neustift inkorporierten) Pfarre St. Sigmund/ Kiens im Pustertal hergestellt und zu einem nicht näher bestimmbaren späteren Zeitpunkt in die Stifts bibliothek Neustift überstellt worden sein.165 Wie bei den beiden obgenannten Antiphonarien-Gradualien ist auch bei Sign. 15063 nicht sicher, ob die Handschrift in Neustift verwendet wurde, da sie weder einen Stempel noch ein Exlibris der Stiftsbibliothek aufweist. Wie bereits bei einer ersten Analyse festgestellt werden kann, ist dieser Codex aus musikwissenschaftlicher Sicht äußerst interessant. Leider ist der ursprüngliche Inhalt an zahlrei chen Stellen verloren gegangen, da viele Blätter radiert und zu einem späteren Zeitpunkt überschrieben wurden. In jenem Teil, der aus dem 15. Jahrhundert stammt, überliefert die Handschrift zahlreiche rhythmische Offizien und Tropen, darunter einen zweistimmigen Tropus zum Benedicamus Procedentem sponsum de thalamo, der auch im bereits genannten Cod. 457 der ULB Tirol vorkommt. Der Stil der sogenannten einfachen Polyphonie scheint sich über lange Zeit in Neustift gehalten zu haben: den handschriftlichen Quellen zufolge bis zur Schwelle zum 19. Jahrhundert. So taucht in einem Psalte rium-Hymnar von 1779, das in der Stiftsbibliothek verwahrt wird, ein „Hymnus secundatus“ auf: Die Hauptmelodie wird von einer in Quarten und Quinten tieferen Stimme begleitet.166 161 162 163 164 165 166 Diese Verwendung ist in Augustiner Chorherren-Handschriften aus Salzburg, Vorau und Seckau belegt; vgl. Prassl, Psallat ecclesia mater (wie Anm. 153) 359 passim. Die von Praßl untersuchten Sequenzen sind auch online unter www.cantusplanus.at verfügbar, zu finden in der Rubrik der Sequenzen in der Liturgie österreichischer Augustiner Chorherren: http://www.cantusplanus.at/de-at/austriaca/prassl/index.php. Siehe AH 54, Nr. 89. Siehe z. B. Cod. 136, Choralhandschrift auf Pergament aus dem Jahr 1762: Sie überliefert eine Reihe von Alleluia-Versen, Gradualien und marianischen Sequenzen, die in der Liturgie eindeutig bis ins späte 18. Jahrhundert gesungen wurden. Ein weiteres Beispiel für die lange Pflege der althergebrachten Gesänge ist zweifellos der genannte „Hymnus secundatus“ im Psalterium-Hymnar von 1779. Engels, Einstimmige liturgische Handschriften (wie Anm. 148) 309–310. Für weitere Informationen hierzu siehe die Beschreibungen der beiden Handschriften im nachfolgenden Katalogteil. Es handelt sich um den Hymnus Iam lucis, in dem eine zweite, in Rot notierte Stimme die in Schwarz notierte Hauptstimme begleitet, ähnlich einem Orgelpunkt (Neustift, Stiftsbibl., Cod. 129,6). 48 Besonderheiten der Erschließung und ihre Ergebnisse Eine weitere Besonderheit dieses Antiphonars stellen die Reihen der Antiphonen zum Magnificat und zum Benedictus für die Sonntage nach Pfingsten dar. Diese stimmen mit der Klosterneuburger Reihe überein.167 Die Reihe der Responsorien zur Nokturn des Totenoffiziums (Bl. 185v–192v) ist hin gegen nirgendwo sonst belegt168, auch nicht in Brixen, und spiegelt daher offensichtlich eine besondere, möglicherweise lokale Tradition wider. Sign. 15063 enthält am 5. Februar außerdem das Offizium der Brixner Diözesanheiligen Ingenuin und Albuin. Der Text des Offiziums entspricht jenem in den handschriftlichen und gedruckten Bre vieren der Diözese Brixen, die hier bezeugte Version mit Notation ist hingegen die zweitälteste und die erste mit diastematischer Notation.169 Die Antiphon zum Magnificat der ersten Vesper Adest dies celebris hat denselben Textanfang wie die Antiphon zum Magnificat des Offiziums für den hl. Augustinus, die in mehreren Handschriften des 13. Jahrhunderts auftaucht, und könnte uns möglicherweise einen Hin weis auf einen Zusammenhang zwischen diesem Offizium und dem Augustiner Chorherrenstift Neu stift geben. Gerade hinsichtlich diverser Zusammenhänge und Beziehungen sind dank musikwissenschaftlicher Detailuntersuchungen in den kommenden Jahren noch Neuerkenntnisse zu erwarten. 3.4 Einbände Ursula Stampfer In der Stiftsbibliothek Neustift sind unterschiedlich gestaltete Einbände auf uns gekommen, die einen Einblick in die Geschichte und Entwicklung des Buchbinderhandwerks erlauben. Zudem lässt sich be stätigen, dass der Einband nicht zwingend mit der Entstehungszeit des Textes übereinstimmen muss bzw. aus diversen Gründen im Laufe der Jahrhunderte ersetzt und erneuert wurde. Für einen möglichst vollständigen Überblick über die Einbände bzw. die Buchbindekunst wäre es notwendig, nicht nur die Einbände der überlieferten Handschriften, sondern auch jene der Archivhandschriften sowie der Inku nabeln zu untersuchen und die Archivalien nach allfälligen Hinweisen auf Stempel- und Rollenkäufe sowie Buchbinder genauestens zu sichten. Die Einbände spiegeln die abwechslungsreiche Geschichte der Neustifter Handschriften wider. Sie legen Zeugnis ab von der Buchpflege in der Romanik, Gotik und insbesondere in der Renaissance, zeigen auf, wie sich die Änderung der räumlichen Verhältnisse in der zweiten Hälfte des 18. Jahrhunderts auch auf die einzelnen Bücher auswirkte, und veranschauli chen die Restaurierpraxis im ausgehenden 20. Jahrhundert. Nur mehr ein kleiner Teil der Codices weist heute noch den ursprünglichen Einband auf. 13 Hand schriften werden von einem gotischen Ledereinband, teilweise schmucklos, teilweise mit Streicheisen linien, teilweise mit Blinddruck verziert, geschützt. Die sieben Renaissance-Einbände sind mehrfach bereits eine Zweitbindung bzw. nicht unmittelbar nach Fertigstellung der Handschrift entstanden. Im Gegensatz zu Archivhandschriften und Urbaren ist nur ein einziger den Bibliothekshandschriften zugeordneter Codex mit einem Makulatureinband versehen: die „Vita Hartmanni“ aus der Zeit um 1200. Ein Codex ist in einen roten Samteinband eingebunden, der wohl aus dem 17. Jahrhundert stammen 167 168 169 Vgl. die Datenbank „Post-Pentecost Cantica Antiphons“ (https://www.uni-regensburg.de/Fakultaeten/phil_ Fak_I/Musikwissenschaft/cantus/). Zur engen Beziehung zwischen den beiden Augustiner Chorherrenstiften Neustift und Klosterneuburg bei Wien siehe allgemeine Einleitung (Abschnitt 1). Vgl. K. Ottosen, The responsories and versicles of the Latin Office of the Dead. Aarhus 1993. Hier werden zu dieser Responsorien-Reihe keine Parallelen aufgezeigt. Das älteste Offizium mit Notation ist im „Calendarium Wintheri“ aus Brixen (Bozen, Staatsarchiv, Cod. 147) erhalten: Die darin enthaltenen Melodien sind allerdings nicht erschließbar, da sie in linienlosen, deutschen adiastematischen Neumen angegeben sind. Eine genaue Beschreibung der Handschrift findet sich in: G. Gabrielli, I manoscritti liturgico-musicali di Bolzano (secoli XIII–XIX) (Bibliotheca Mediaevalis 3). Lucca 2015, 3–8. Besonderheiten der Erschließung und ihre Ergebnisse 49 dürfte. Der weitaus größte Teil der Handschriften (64) zeigt heute einen Bibliothekseinband im Stile eines Halbfranzeinbandes aus der Zeit um 1780, samt Titelaufschrift in Golddruck am Rücken. Dass das sogenannte Posch-Missale wohl im selben Zeitraum mit einem schlichten Kalbsledereinband ge bunden und Vorder- und Hinterdeckel nicht wie bei den anderen mit einem schwarz-braun gesprenkel tenPapier überzogen wurden, hängt wohl mit dessen bereits damals erkannter Bedeutung zusammen. Im Zuge von Restaurierungen erhielten vier Handschriften in der zweiten Hälfte des 20. Jahrhunderts einen neuen Einband. Die älteste heute in der Stiftsbibliothek aufbewahrte Handschrift (Cod. 495) stammt aus dem 10. Jahrhundert und ist mit einem schlichten Einband, der wohl nur kurze Zeit nach Abschluss des Textes angefertigt worden sein dürfte, versehen: Holzdeckel, die mit einem schmucklosen, im Laufe der Jahrhunderte stark nachgedunkeltem Schweinsleder überzogen sind. Eine genaue Datierung und Lokalisierung des Einbands ist aufgrund mangelnder Sekundärhinweise nicht möglich. Die Titelauf schrift am Vorderdeckel aus dem 12. Jahrhundert sowie die noch erhaltenen hervorstehenden Dorne der einstigen Schließen machen deutlich, dass das Buch ursprünglich liegend aufbewahrt wurde. Der blau gefärbte Rücken mit einer weißen Titelaufschrift – es lässt sich nicht feststellen, wann dieser Eingriff erfolgte170 – bezeugt nicht nur die spätere Aufstellung in Regalen, sondern auch eine einstige Zugehö rigkeit zu einer Teilbibliothek. Solche blau gefärbten Rücken weisen nicht nur einige weitere Hand schriften auf (Cod. 142, 194, 440, 726 und Nr. 139 [olim Sign. 14819] sowie Innsbruck, ULB Tirol, Cod. 688, 731), sondern mehr als 300 Druckwerke. Wozu diese Blaufärbung diente, ist bis dato noch nicht geklärt. Möglicherweise waren diese Bücher zu einer bestimmten Zeit Teil einer Art „Spezial bibliothek“ im Haus; nicht zuletzt aufgrund der Tatsache, dass sich unter diesen Bänden auch das „Liber anniversariorum Neocellensis“, das „Necrologium Neocellense“ sowie ein besonders schön ausgeschmücktes Neustifter Rituale befinden, könnte man an eine Aufstellung in der Prälatur denken. Die zweite, wohl etwas ältere Form der besonderen Rückengestaltung, nämlich ein an den Bünden und im zweiten Rückenfeld schwarz gefärbter Rücken171, lässt sich bei den Handschriften nicht nachweisen. Bei den erhalten gebliebenen gotischen Einbänden aus dem 15. Jahrhundert ist der Lederbezug, der bisweilen rot gefärbt ist, vielfach nur sehr spärlich mit Blindlinien verziert. Einige Einbände weisen aber auch eine reiche Dekoration mit Einzelstempeln auf, die meist in von mehrfachen Streicheisen linien umrandeten Feldern platziert sind. Diese Stempel, diverse Blüten, Lilien, Adler, Einhorn, Spruch bänder Maria u. a., konnten bislang keinem konkreten Buchbinder bzw. keiner Werkstatt zugeordnet werden; eine Lokalisierung in den süddeutschen, wenn nicht gar Tiroler Raum ist anzunehmen. Eine genauere Erforschung derselben, verbunden mit einer umfangreichen Archivrecherche bleibt bis auf Weiteres noch ein Desiderat. Reicher verziert sind lediglich die Einbände zweier Handschriften mit liturgischem Inhalt. Die Antiphonarien und Gradualien (Nr. 139 [olim Sign. 14819] und Nr. 940) wurden in den 1490er Jahren von Heinrich Edlinger geschrieben, wohl im direkten Umfeld des Augustiner Chorherrenstiftes, ebendort dürfte auch die Bindung zu lokalisieren sein. Hierfür spricht insbesondere der Einzelstempel Osterlamm mit Fahne in Kreis, der auch beim Einband von Cod. 11 des Stiftsarchivs Verwendung fand (dat. 1580: Rechnungsbuch des Propstes Augustin Schabl), dort in Kombination mit Rollen (Neuhauser R 48 und R 56, Gruppe 7172), die eindeutig nach Neustift weisen. Der Einzelstempel dürfte also möglicherweise zu einem Formenschatz gehört haben, der im Kloster selbst aufbewahrt und 170 171 172 Diese Färbung dürfte mit einer Neuaufstellung einhergegangen sein, die möglicherweise im Zuge der Errichtung der akademisch-theologischen Hauslehranstalt in der zweiten Hälfte des 17. Jahrhunderts, der Neugestaltung der Prälatur in der ersten Hälfte des 18. Jahrhunderts oder der Neugestaltung der Stiftsbibliothek in der zweiten Hälfte des 18. Jahrhunderts erfolgte. Neuhauser, Wissenschaftspflege (wie Anm. 23) 98. W. Neuhauser, Die Rolleneinbände des Buchdruckers und Buchbinders Gallus Dingenauer: Meister G-D, in: G. Colin (Hrsg.), De libris compactis: miscellanea (Studia Bibliothecae Wittockianae 1). Bruxelles 1984, 133–249, hier 196–199. 50 Besonderheiten der Erschließung und ihre Ergebnisse den jeweiligen Buchbindern für Auftragsarbeiten173 zur Verfügung gestellt wurde. Wenn wir mit der Annahme, dass Nr. 139 (olim Sign. 14819) und Nr. 940 Ende des 15./Anfang des 16. Jahrhunderts, also kurz nach Erstellung des Textes gebunden wurden, richtigliegen, so wäre dieser Stempel fast 100 Jahre im Einsatz gewesen. Zwei weitere Einbände aus dem 15. Jahrhundert verdienen eine besondere Erwähnung: Cod. 4, der ein Repertorium beider Rechte von Iohannes Calderinus enthält und vom Brixner Domherrn Johann Ebner174 nach seinem Tod 1461 dem Stift überlassen wurde, weist am Hinterdeckel oben mittig die Spur eines ursprünglich dort befestigten runden Beschlags auf. Es ist also anzunehmen, dass es sich hierbei ursprünglich um ein Kettenbuch handelte.175 Völlig anderer Natur ist hingegen der Einband der um 1200 entstandenen „Vita beati Hartmanni“ (Cod. 293), die im 15. Jahrhundert neu gebunden und mit einer – ansonsten in Neustift insbesondere für Archivhandschriften üblichen – Makulatur umschlagen wurde: Das beidseitig beschriebene und mit gotischer Choralnotation versehene Pergament-Doppelblatt aus einem Antiphonar, aufgrund der Schrift ins 14. Jahrhundert zu datieren, das offensichtlich seiner ursprünglichen Verwendung entledigt worden war, ist über dünne Pappdeckel gespannt. Während die erhaltenen gotischen Einbände größtenteils wohl zeitnah zur Herstellung bzw. zum Erwerb der Handschriften entstanden, belegt das Gros der Einbände aus späterer Zeit eine Welle an Neubindungen, die wohl in engem Zusammenhang mit dem steigenden Interesse an Büchern und Biblio theken in Tirol in der zweiten Hälfte des 16. Jahrhunderts steht.176 Durch das Aufblühen des Buchhan dels im Allgemeinen und den verstärkten Bücherkauf durch Klöster im Zuge der Gegenreformation erhielten nicht nur die Buchhändler, sondern insbesondere auch die Buchbinder vermehrt Aufträge. Die Einbände aus jener Zeit sind reich mit Einzelstempeln, Rollen und Platten verziert, teilweise in Blind druck, teilweise in Golddruck, bei mehreren Büchern ist auch der Schnitt golden gefärbt und punziert. Eine systematische Erforschung sämtlicher in Tirol wirkender spätmittelalterlicher und frühneuzeitli cher Buchbinder und deren Werkstätten ist zwar noch ausständig, doch dank der grundlegenden Arbeit von Walter Neuhauser zu den im Tiroler Raum auffindbaren Einbänden aus der zweiten Hälfte des 173 174 175 176 Bislang konnte kein Hinweis auf einen stationären Buchbinder im 15. und beginnenden 16. Jahrhundert in Neustift gefunden werden; man kann wohl von Wanderbuchbindern ausgehen. Möglicherweise identisch mit Johannes Ebner aus Krems, der 1414 an der Universität Wien immatrikuliert war; vgl. L. Santifaller, Das Brixner Domkapitel in seiner persönlichen Zusammensetzung im Mittelalter (Schlern-Schriften 7). Innsbruck 1924, 298–299; Die Matrikel der Universität Wien I: 1377–1450, bearb. von F. Gall (Publikationen des Instituts für Österreichische Geschichtsforschung VI, 1). Graz, Köln 1956, 104. In Neustift lassen sich ansonsten keine Hinweise auf Kettenbücher finden. Ob diese gerade im Umfeld von Schultätigkeit weit verbreitete Sicherungspraxis tatsächlich in Neustift nicht üblich war oder ob sich lediglich keine entsprechenden Einbände erhalten haben, muss offenbleiben. Möglicherweise belegt gerade der Einband, dass Cod. 4 aus dem Besitz eines Brixner Domherrn ursprünglich an der seit dem 10. Jahrhundert urkundlich belegten Domschule Brixen verwendet worden war. Hier sei insbesondere an den bibliophilen Erzherzog Ferdinand II. (1529–1595) und dessen umfangreiche Bibliothek auf Schloss Ambras erinnert; vgl. I. Purš, Die Bibliothek Erzherzog Ferdinands II. auf Schloss Ambras, in: S. Haag, V. Sandbichler (Hrsg.), Ferdinand II. 450 Jahre Tiroler Landesfürst. Jubiläumsausstellung. Innsbruck, Wien 2017, 99–106; Das Inventarium der Ambraser Sammlungen aus dem Jahre 1621. T. 2: Bibliothek, hrsg. und kommentiert von A. Auer, in: Jahrbuch des Kunsthistorischen Museums Wien, Bd. 2. Wien 2001, 281–345; A. Auer, E. Irblich, Natur und Kunst. Handschriften und Alben aus der Ambraser Sammlung Erzherzog Ferdinands II. (1529–1595). Ausstellungskatalog. Wien 1995. Als Landesfürst kam ihm sicherlich Vorbildfunktion zu; in der zweiten Hälfte des 16. Jahrhunderts entstanden vermehrt private Bibliotheken. Über die wohl umfangreichste verfügte die Familie der Freiherren von Wolkenstein-Rodenegg; vgl. U. Stampfer, Adelige Lesekultur um 1600 am Beispiel der WolkensteinRodenegg, in: G. Pfeifer, K. Andermann (Hrsg.), Die Wolkensteiner. Facetten des Tiroler Adels in Spätmittelalter und Neuzeit (Veröffentlichungen des Südtiroler Landesarchivs 30). Innsbruck 2009, 323–334; A. Dörrer, Die Bibliothek des Freiherrn Christoph von Wolkenstein auf Schloss Rodenegg. Ein adeliges Gegenstück zur landesfürstlichen Ambraser Sammlung. Zentralblatt für Bibliothekswesen 57 (1940) 18–43; Ders., Tirolisches aus der Bücherei des Freiherrn Christoph von Wolkenstein auf Rodenegg, in: Tirolensia zum 80. Geburtstag Konrad Fischnaler’s (Schlern-Schriften 30). Innsbruck 1935, 21–26. Besonderheiten der Erschließung und ihre Ergebnisse 51 16. Jahrhunderts und der Erfassung von Schmuckformen177 können mehrere der Neustifter Einbände bereits bekannten Buchbindern bzw. Buchbindewerkstätten zugewiesen werden. In Neustift nament lich fassbar ist allerdings bis dato nur Narziß Schuechl. Er stammte aus Augsburg, dürfte kurz nach 1560 nach Innsbruck gekommen sein und wurde ebendort am 18. März 1568 als Bürger aufgenommen. Über sein Leben sind wir nur spärlich unterrichtet; über seine Tätigkeit als Buchbinder informieren uns die erhalten gebliebenen Einbände für die Bibliothek des Jesuitenkollegs in Innsbruck, das Prämons tratenser Chorherrenstift Wilten, das Zisterzienserstift Stams, die Franziskanerklöster in Innsbruck und in Schwaz sowie für Privatpersonen. Man kann also auf eine gute Auftragslage schließen. Einträge im Raitbuch178 des Neustifter Propstes Augustin Distelfink (1585–1589) belegen, dass er in der Zeit von 1585 bis 1589 auch mehrere Bücher für das Augustiner Chorherrenstift eingebunden hat.179 Zumal er in den Rechnungen als Narzischg puechbinter zu Brichsen bezeichnet wird, ist anzunehmen, dass er sich immer wieder für längere Zeit in der Bischofsstadt aufhielt und dort über eine eigene Buchbinderwerk stätte verfügte.180 Auf der Basis der bereits erschlossenen und im Detail untersuchten Handschriften im Tiroler Raum, insbesondere der Stiftsbibliothek Wilten, der Priesterseminarbibliothek Brixen und der ULB Tirol, sind ausgehend von den verwendeten Stempeln und Rollen – sie sind allesamt nicht sig niert – Rückschlüsse auf seine Arbeiten möglich. Die bei den Einbänden der Cod. 353, 654, 726, Sign. 15063 sowie des ersten Bandes des Neustifter Gradualbuches (bei Band 2 ist der Einband zur Gänze Ende des 20. Jahrhunderts erneuert worden) verwendeten Rollen und Platten lassen sich der von Neu hauser definierten Gruppe 1 und 2 sowie 7 zuordnen, die miteinander sehr eng verknüpft sind.181 Trotz Schwierigkeiten bei der Abgrenzung der einzelnen Gruppen untereinander können all diese Dekor elemente mit größter Wahrscheinlichkeit Schuechl bzw. seiner Werkstatt zugewiesen werden. Auf grund der großen Ähnlichkeit des Einbandes von Cod. 405 mit jenen von Cod. 353 und 654 (vgl. ins besondere auch die Schließen) kann auch jener Band Schuechl und seiner Werkstatt zugeordnet werden. Während die Einbände der Gruppe 1/2 laut Neuhauser in den Zeitraum 1568–70 datieren, stammen jene mit Dekorelementen aus Gruppe 7 aus der Zeit zwischen ca. 1570 und nach 1600. 177 178 179 180 181 Neuhauser, Dingenauer (wie Anm. 172). Im Kontext von Tiroler Bucheinbänden dürfen zudem folgende Arbeiten nicht unerwähnt bleiben; sie wurden hinsichtlich des vorliegenden Neustifter Bestandes konsultiert, es konnten allerdings keine Übereinstimmungen festgestellt werden: W. Neuhauser, Eriber. Ein Beitrag zum süddeutschen und Tiroler Bucheinband des 15. Jahrhunderts, in: A. De Coster, C. Sorgeloos (Hrsg.), Biblio philies et reliures. Mélanges offerts à Michel Wittock. Bruxelles 2006, 317–373; E. Ramminger, Augsburger Rolleneinbände des 16. Jh.s an der Universitätsbibliothek Innsbruck, bibliothekarische Hausarbeit. Innsbruck 1987 (maschinschr.); S. Sepp, Spätgotische Kölner Einbände aus der ehemaligen Haller Jesuitenbibliothek in der Universitätsbibliothek Innsbruck. Codices manuscripti 6 (1980) 89–111; W. Neuhauser, Erfurter Blind stempeleinbände an der Universitätsbibliothek Innsbruck. Codices manuscripti 5 (1979) 8–24; O. Mazal, Gotische Einbände aus der Kartause Schnals. Gutenberg-Jahrbuch (1973) 423–428. Neustift, Stiftsarchiv, Cod. 26. 1585: 27. Mai: Dem Narzischg puechbinter zu Brichsen fur das rodl püechl und zway andere püecher einzubinden bezalt 1fl 20 kr.; 22. August: Den Narzischgg puechpintter zway postille lassen einpinden geben 2 fl. 1586: 10. August: Am 10 dito dem Narcis puechpinter zu Brixen umb willen etlich püecher einzubinden bezalldt 3 fl. 28 kr. 1587: 25. Jänner: Am 25 dits dem puechpinter zu Brixen das er 15 tag bei mir hat eingepunden darvir bezalt ich ime 4 fl. 1588: 13. Jänner: Am 13. dem buechpinter zu Brixen, dem maister Narziß, der mir im gotshauß etlich corpuecher unnd anndrer puecher eingebunden hat, fur 32 tag bezalt 8 fl. 52 kr.; 12. Mai: Dem Narciß puechpinter fur 2 misell unnd ain corporaetaschen zu uberziechen zalt 4 fl 30 kr.; 18. September: Am 18. dem Narciß buechpindter fur ain prefier einzupindten geben 24 kr.; 9. November: Eodem die dem Narciß puechpindter zu Brixen das er mir im gotshauß hat 7 tag einpundten bezalt ain tag 16 kr – 1 fl. 52 kr. Die bisherige Annahme, dass er erst 1588 seine Werkstätte nach Brixen verlegt habe, dürfte hiermit entkräftet sein; möglicherweise ist Narziß Schuechl in jenem Jahr lediglich vollständig nach Brixen übersiedelt. Vgl. Neuhauser, Dingenauer (wie Anm. 172) 156; F. Waldner, Quellenstudie zur Geschichte der Typographie in Tirol. Innsbruck 1888, 103. Sowohl Gruppe 2 als auch Gruppe 7 sind wohl als Fortsetzung der Gruppe 1 anzusehen (Neuhauser, Dingenauer [wie Anm. 172] 196–197). 52 Besonderheiten der Erschließung und ihre Ergebnisse Als für Neustift wohl bedeutendste Rolle ist jene hervorzuheben, die das Stiftswappen, aufgeteilt auf drei einander abwechselnde Einzelmedaillons (vier Spitzen der Herren von Säben, Tau-Kreuz und Patriarchenkreuz), zeigt. Möglicherweise wurde diese Wappenrolle, die sich auch am Einband des Pla citum aus dem Jahr 1579 sowie am Rechnungsbuch von Propst Augustin Schabl (1569–1581) befindet, von eben jenem Propst in Auftrag gegeben.182 Ein Wappensupralibros, das einen Erzengel (Michael?) mit einem Schild mit zwei aufsteigenden Löwen und damit den Wappenschild dieses Propstes zeigt (Neuhauser P 3), belegt anschaulich die bibliophile Haltung Schabls. Mit dieser Platte sind die beiden Gebetbücher des Klausner Stadtrichters Kaspar Neuhauser geschmückt (Cod. 353, 654), die zu einem nicht näher definierten Zeitpunkt in den Besitz des Klosters übergingen und unter Propst Schabl neu gebunden wurden. In der ULB Tirol lassen sich zudem weitere drei Bände mit dieser Platte nachweisen (Ink. 106 E 1, Ink. 106 E 2: Breviarium Salisburgense, Pars hiemalis und Pars aestivalis. Nürnberg: Georg Stuchs 1497 [GW 5444]; Sign. 100306: H. Pantaleon, Teutscher Nation Heldenbuch … erst lich … in Latein zusammen gezogen … jetzmalen aber von dem ersten authore selbs verteütschet … Basel: Nicolaus Brylinger, 1567–1570).183 Während das Wappensupralibros selbstredend später keine Verwendung mehr fand, taucht die Wappenrolle auch noch auf einem Druck aus dem Jahr 1609 auf.184 Diese lebhafte (Neu-)Bindetätigkeit Ende des 16./Anfang des 17. Jahrhunderts lässt sich auch in anderen Klöstern beobachten: So wurden im Prämonstratenserstift Wilten zwischen ca. 1590 und 1610 von Leonhard Schuechl – möglicherweise ein Sohn von Narziß Schuechl – zahlreiche Handschriften und Inkunabeln neu gebunden.185 Bei den allermeisten Handschriften wurde in der zweiten Hälfte des 18. Jahrhunderts der mittel alterliche oder frühneuzeitliche Einband entfernt und der Einheitlichkeit wegen durch einen schlichten Bibliothekseinband ersetzt. Diese Neubindungen stehen zweifelsohne allesamt im Zusammenhang mit dem Neubau des Bibliothekssaales und der Neuaufstellung der Bibliothek unter Propst Leopold de Zanna. Auch zahlreiche Druckwerke wurden mit demselben Einband versehen; vielfach wurden hier bei aber die ursprünglichen Deckel beibehalten und lediglich der Rücken erneuert, um so dem Biblio thekssaal ein möglichst harmonisches Bild zu verleihen. Dieser Halbfranz- bzw. Halbledereinband zeichnet sich durch Pappdeckel, die mit schwarz gesprenkeltem braunem Papier überzogen sind, sowie einen braunen Lederrücken aus. Mit demselben braunen Leder sind auch die Ecken jeweils verstärkt. Die einfachen Bünde am Rücken werden von schlichten Bordüren in Golddruck gesäumt, in den Feldern zwischen den Bünden ist M. S. (Manuscriptum) in Golddruck festgehalten, zudem ist meist ein Werk titel oder vereinzelt auch der Autor des Textes angegeben. Werke von geringerem Umfang, wie z. B. Cod. 578, 579, 597, 691, 694, 754, 755, 756 und 780, wurden hingegen mit demselben Papier, mit dem bei anderen Bänden die Pappe umschlagen wurde, lediglich broschiert. Auffallend ist die Tatsache, dass im Rahmen dieser Neubindungen mehrere Sammelbände in ihre Einzelbestandteile aufgelöst wurden, von denen jeder einen entsprechenden Halbfranzeinband erhielt.186 Aus den Rechnungsbüchern des 182 183 184 185 186 Aufgrund dieser beiden Archivhandschriften, die wohl kaum eine Neubindung wenige Jahre nach der Erstellung erfahren haben dürften, ist die von Neuhauser geäußerte These, dass die Wappenrolle auf eine Bindung unter Propst Jakob Fischer (1589–1621) schließen ließe, wohl kaum haltbar. Die Tatsache, dass das Tau-Kreuz am unteren Ende einen Sporn habe – laut Neuhauser ein Hinweis darauf, dass die Rolle in jene Zeit zu datieren sei, zumal Propst Fischer der letzte war, unter dem das Tau-Kreuz diese Form aufwies (vgl. Neuhauser, Wissenschaftspflege [wie Anm. 23] 99) – spricht meines Erachtens nicht gegen eine frühere Datierung. Neuhauser, Dingenauer (wie Anm. 172) 214. Neuhauser, Wissenschaftspflege (wie Anm. 23) 99. Neuhauser, Dingenauer (wie Anm. 172) 141, 160, 189. So bildeten die ursprünglich selbständigen Handschriften aus dem 14. und 15. Jahrhundert Cod. 158, 83, 82, 183, 161, 182 und 162 sowie Cod. 157, 159, 168, 163, 181 und 164 (in eben dieser Reihenfolge) seit dem 15. Jahrhundert bis dahin jeweils eine Einheit. Heute werden Teile dieser einstigen Sammelhandschriften mit durchgehender Foliierung und gemeinsamem Inhaltsverzeichnis aus dem 15. Jahrhundert in der Stiftsbibliothek Neustift, andere an der ULB Tirol (Cod. 82 sowie Cod. 157, 159 und 164) aufbewahrt. Diese Bände führen die Notwendigkeit bzw. Sinnhaftigkeit einer virtuellen Rekonstruktion einstiger Bestände deutlich vor Augen. Besonderheiten der Erschließung und ihre Ergebnisse 53 Propstes187 geht hervor, dass diese Neubindungen bereits Anfang der Sechzigerjahre des 18. Jahr hunderts begonnen wurden188 und sich bis ins Jahr 1778189, also eben jenes Jahr, in dem auch der Neu bau des Bibliothekssaales abgeschlossen werden konnte, fortsetzten. Im Zuge dieser Neubindungen dürfte auch das Exlibris der Stiftsbibliothek in Holzdrucktechnik mit Zierrahmen, das bis heute viele Handschriften und Drucke schmückt, eingeklebt worden sein. Insgesamt hat der bzw. haben die Buch binder in diesen Jahren 464 Gulden, 59 Kreuzer verdient. Wer diese Arbeiten durchgeführt hat, ist leider nicht erwähnt; zumal sie aber Eingang in die Rechnungsbücher gefunden haben, kann davon ausgegangen werden, dass kein Chorherr bzw. Klostermitglied diese Neubindungen vorgenommen hat. Warum nicht ausnahmslos alle Einbände erneuert bzw. umgestaltet wurden, muss offenbleiben; eine mögliche Erklärung könnte der Aufbewahrungsort sein. Die Handschriften, die auch heute noch einen mittelalterlichen oder frühneuzeitlichen Einband aufweisen, sind großteils liturgischen Inhalts und wurden möglicherweise nicht im neuen Bibliothekssaal aufgestellt, sondern wurden im näheren Um feld der Kirche aufbewahrt. Cod. 142, der neben der Ordensregel des hl. Augustinus und der Auslegung derselben durch Hugo de Sancto Victore das Anniversarium und Necrologium des Stiftes enthält, dürfte hingegen im Archiv geblieben sein und daher seinen schlichten Renaissance-Einband behalten haben. Sieben Handschriften erhielten im Rahmen einer Neubindung bzw. Restaurierung in den ver gangenen Jahrzehnten (wohl um 1980)190 einen völlig neuen Einband. Wie den – meist am Spiegel des Hinterdeckels – aufgeklebten Etiketten zu entnehmen ist, wurden Cod. 453, 483, 532, 550191, 614 und der zweite Band des Zollner-Graduales in der Restaurierwerkstätte des Benediktinerinnenstiftes Santa Maria di Rosano in Rignano sull’Arno bei Florenz neu gebunden. Cod. 776 enthält kein Etikett; der eingebundene schmale Papierstreifen mit italienischer Beschriftung lässt vermuten, dass auch er seinen neuen Einband in einer italienischen Restaurierwerkstätte erhielt. Auch der erste Gradualband und Cod. 123 wurden in Santa Maria di Rosano neu gebunden und deren Einbände unter Verwendung des ursprünglichen Lederbezugs restauriert. In den letzten Jahren wurden die Restaurierungen im Sinne einer bestmöglichen Konservierung der Handschriften fortgesetzt. Die Arbeiten wurden von Buchrestauratoren aus Süd- und Nordtirol durchgeführt.192 187 188 189 190 191 192 Neustift, Stiftsarchiv, Cod. 24. Der erste entsprechende Eintrag reicht in den Juni 1761 zurück: an büchern pro Bibliotheca item büchbindern 92 fl. (Neustift, Stiftsarchiv, Cod. 24, S. 141). Neustift, Stiftsarchiv, Cod. 24, S. 188 ist unter Ausgaben in Jenner 1778 vermerkt: dem büchbündter laut conto 102 fl., 9 kr. Ein Restaurierbericht konnte leider für keine dieser Handschriften ausfindig gemacht werden. Die ursprünglichen, wohl aus dem 15. Jahrhundert stammenden, mit einem rot gefärbten Leder überzogenen Holzdeckel sind erhalten und werden getrennt aufbewahrt (ursprünglich im Stiftsarchiv, heute liegen sie bei den Fragmenten). Ein Dank geht an Simon Terzer für die Übermittlung von Bildmaterial. Auf beiden Holzdeckeln ist innen jeweils der Abklatsch einer Urkunde zu sehen. Auf dem ursprünglichen Vorderdeckel – darauf lassen die an der Universitätsbibliothek in Innsbruck notierten Signaturen (in Tinte 550 und II […] sowie in Bleistift II3 D 8) schließen – ist der Abklatsch einer quer eingeklebten, 22zeiligen, seitlich rechts beschnittenen Urkunde mit klarem Brixenbezug zu erkennen, u. a. ist von der Pfarre St. Michael die Rede. Besonders interessant ist die Nennung des Domherrn Johannes Ebner (siehe Anm. 174), der dem Kloster Neustift nach seinem Tod 1461 eine Handschrift (Cod. 4) testamentarisch überlassen hat. Auch in der 33zeiligen Urkunde am ursprünglichen Hinterdeckel begegnet uns Johannes Ebner. Ob dies ein Hinweis darauf ist, dass die Handschrift auf demselben Weg wie Cod. 4 Eingang in die Neustifter Stiftsbibliothek gefunden hat, muss offenbleiben. Sofern Informationen zu Restaurierungen vorliegen, wurde dies in den Katalogisaten vermerkt. 55 4. Bisherige Erschließung des Handschriftenbestandes Ursula Stampfer Die vorliegende Publikation stellt die erste systematische Erfassung und wissenschaftliche Erschlie ßung sämtlicher in Neustift aufbewahrter mittelalterlicher Handschriften dar. Bislang lag lediglich ein nach Signaturen geordneter Zettelkatalog vor, der an der ULB Tirol unter Direktor Friedrich Leithe (1868–1874) bzw. Ludwig von Hörmann (1882–1902) erstellt wurde. Bei der damaligen Beschreibung der Handschriften stützte man sich nur auf die in ihnen selbst enthaltenen Daten und unternahm keine weiteren Literaturrecherchen u. ä. zur Identifizierung von Texten, sodass die Angaben vielfach sehr un genau und unvollständig sind.193 Wohl im Zuge der Rückgabe von knapp 100 Handschriften an Italien wurden aus dem Konvolut an Beschreibungskärtchen aller Handschriften an der ULB Tirol die Katalo gisate zu den Neustifter Beständen zu einer separaten Einheit zusammengefasst. Sie wurden zu einem späteren Zeitpunkt – handschriftlich findet sich die Ergänzung [1973] – nochmals in die Hand genom men und um einige maschinschriftliche Karteikärtchen zu den Archivhandschriften des Stiftes ergänzt. Bei den Buchhandschriften selbst wurden nur sehr wenige Ergänzungen gemacht; u. a. wurde vermerkt, ob ein Mikrofilm vorliegt oder nicht. Ein handschriftlicher Hinweis fehlt am linken oberen Blattrand bei jenen Handschriften, die an der ULB Tirol verblieben sind, lässt darauf schließen, dass diese Addenda des Zettelkatalogs in Neustift erfolgt sein dürften. Kopien des Zettelkatalogs mit dem Titel „Die historischen Handschriften des Archivs bzw. der Bibliothek des Augustiner Chorherrenstiftes Neustift“ befinden sich heute sowohl in der Stiftsbibliothek Neustift als auch an der ULB Tirol; unter https://manuscripta.at/kataloge/IT/5000/histHss/histHss.pdf kann er zudem online eingesehen werden. H. Martin Peintner erstellte 1972 einen Zettelkatalog zu jenen Handschriften, die aus Platzgründen nicht wie die wertvollen hist. Codices im Archiv aufbewahrt werden konnten. Die in diesem maschin schriftlichen Verzeichnis aufgelisteten Handschriften sind – mit Ausnahme von Nr. 139 (olim Sign. 14819) – allesamt jüngeren Datums. H. Theobald Innerhofer erstellte ein Verzeichnis aller im Archiv aufbewahrten Bibliotheks- und Archivhandschriften; seine Ausführungen zu den Bibliothekshandschriften reichen aber nicht über die Notizen des früheren Zettelkatalogs hinaus. Ein wissenschaftlicher Katalog lag lediglich für die illuminierten Handschriften vor: Der Wiener Kunsthistoriker Hermann Julius Hermann (1869–1953) veröffentlichte 1905 eine umfassende Arbeit zu den illuminierten Handschriften in Tirol (siehe Anm. 137 und Literatur). Die Neustifter Bestände, damals noch großteils in Innsbruck verwahrt, sind zum Teil unter den dortigen Beständen, zum Teil im Kapitel zu Neustift selbst zu finden. In den folgenden Jahren folgten mehrere Einzelstudien zu aus gewählten Handschriften; seit jeher ziehen insbesondere die reich mit Buchmalerei ausgestatteten Werke die Aufmerksamkeit der Forschung auf sich. In jüngerer Zeit verfasste vor allem H. Martin Peintner mehrere Publikationen zum Neustifter Skriptorium. 1979 fand schließlich die siebte Tagung der österreichischen Handschriftenbearbeiter teilweise in Neustift statt – wohl ein deutliches Zeichen für die Bedeutung, die der dortigen Handschriftensammlung einerseits im Allgemeinen, andererseits aber wohl auch als zentralem Komplementärbestand zu den Innsbrucker Handschriften im Speziellen 193 Katalog ULB Tirol I (wie Anm. 100) 21. 56 Erschließung des Handschriftenbestandes beigemessen wird. Letztere wurden seit Mitte der 1970er Jahre systematisch nach den Richtlinien der ÖAW tiefenerschlossen, in Neustift sollten noch mehr als drei Jahrzehnte vergehen, ehe die wissen schaftliche Katalogisierung in Angriff genommen werden konnte. Viele Initialen und Miniaturen von Neustifter Handschriften sind in diversen Publikationen ab gebildet. Die erste systematische Aufnahme sämtlicher mittelalterlicher Handschriften wurde durch Mitarbeiter der Hill Monastic Manuscript Library (HMML) in Collegeville (Minnesota) in den 1960er Jahren durchgeführt; Kopien der Mikrofilme liegen im Stift. Kurze Zeit später erfolgte eine weitere Mikroverfilmung durch das staatliche Denkmalamt in Verona; auch hiervon werden Kopien in Neustift aufbewahrt. Nach Abschluss der Katalogisierungstätigkeit ließ das Stift alle im vorliegenden Katalog berück sichtigten Handschriften digitalisieren. Die Digitalisate sind unter www.manuscripta.at einsehbar. Die datierten und/oder mit einer Schreibernennung versehenen Neustifter Handschriften finden zudem Berücksichtigung im Band zu den „Manoscritti datati della provincia di Bolzano“. Dieser Katalogist derzeit in Vorbereitung und wird in der Reihe „Manoscritti datati d’Italia“ erscheinen.194 194 Vgl. hierzu http://www.manoscrittidatati.it. Basierend auf den Diplomarbeiten von Cristina Stefani (I manoscritti datati della Provincia di Bolzano. Bolzano, Biblioteca del Monastero di Muri-Gries. Bressanone, Biblioteca del Seminario Maggiore. Novacella, Archivio dell’Abbazia di S. Maria, Tesi di laurea. Trento 1995–1996) und Sissi Mattiazzi (Un esempio di biblioteca francescana: la Biblioteca dei Frati Minori di Bolzano, Tesi di laurea. Padova 2011–2012) werden hierin von Leonardo Granata und Ursula Stampfer sämtliche datierte Handschriften in Südtiroler Bibliotheken erfasst. 57 5. Zur Anlage des Katalogs Claudia Schretter-Picker 5.1 Zur Erschließungsmethode Mit der vorliegenden Publikation wird im Rahmen der im Umfeld der Abteilung Schrift- und Buch wesen, Institut für Mittelalterforschung, der ÖAW angesiedelten Projekte zur wissenschaftlichen Erschließung von Handschriften eine bisher nur in Ansätzen erprobte Methode der Handschriften beschreibung intensiviert, nämlich die der arbeitsteiligen Bearbeitung der einzelnen Handschriften. Damit soll dem Umstand Rechnung getragen werden, dass es sich beim vorliegenden Handschriften bestand der Neustifter Stiftsbibliothek um einen Komplementärbestand zu den bereits bearbeiteten Handschriften Neustifter Provenienz in der ULB Tirol handelt. Erst eine intensive, bestandsüber greifende Untersuchung der verschiedenen Aspekte der jeweiligen Handschrift – Zusammensetzung, Beschreibstoff, Schrift, Einband, Ausstattung, Notation, Geschichte, Inhalt – liefert die Basis für die virtuelle Zusammenführung dieses heute getrennten Bestandes. Für die im Neustifter Bestand reichlich vertretenen Handschriften mit höherrangigem Buchschmuck erschien eine nach den Richtlinien und der Terminologie für die Handschriftenbeschreibung in Österreich erfolgende nur knappe Beschreibung als nicht zielführend, zumal ein Spezialkatalog der illuminierten Handschriften für den Tiroler Raum nicht in absehbarer Zeit zu erwarten ist. Auch die Handschriften mit Notation lohnen eine intensivere Betrachtung. Trotz des hohen zeitlichen, personellen und finanziellen Aufwandes wurde also nicht nur an der Methode der Tiefenerschließung festgehalten, sondern diese durch leichte Adaptionen sogar in tensiviert. Der dadurch lukrierte Mehrwert wurde in den Gutachten und Evaluationen zum Antrag für die Finanzierung dieses Erschließungsprojektes durchwegs positiv bemerkt und die Methodik begrüßt. Noch vor der eigentlichen Katalogisierungsarbeit waren bereits die Daten zu allen Neustifter Handschriften aus dem Alten Zettelkatalog an der ULB Tirol und den Anhängen der bereits erschienenen gedruckten Kataloge zu den heute an der ULB Tirol befindlichen Neustifter Handschriften gesammelt worden. Diese betrafen einerseits summarische Angaben zum Inhalt der Handschriften, aber auch Hinweise auf Beschreibstoffe, Format und Datierung. In einem zweiten Schritt wurden Verweise zu den jeweiligen Handschriften aus Handschriften repertorien und Sekundärliteratur gesammelt. Danach erfolgte eine Autopsie aller Handschriften, wobei in rudimentärer Form Informationen zu äußeren und inhaltlichen Merkmalen erfasst wurden. Diese wurden mit den bereits vorhandenen Daten kumuliert und für die Weiterbearbeitung vorbereitet. Auf dieser Basis konnten Handschriftengruppen gebildet werden, differenziert nach Beschreibstoff, Datierung, Ausstattungselementen, Notation sowie inhaltlichen Kriterien. Damit war die Voraussetzung für eine arbeitsteilige Bearbeitung des Bestandes durch ein Team von Spezialistinnen und Spezialisten geschaffen. Um den hohen Ansprüchen der detaillierten Beschreibungsrichtlinien Genüge leisten zu können und dennoch zeitökonomische Bedingungen zu erfüllen, wurde die Beschreibung der äußeren Merk male der Handschriften zunächst durch Autopsie vorgenommen. Die zeitaufwändige Identifikation der 58 Zur Anlage des Katalogs Texte und der Nachweis von Parallelüberlieferungen erfolgten teils anhand von Mikrofilmen und teils mit Arbeitsdigitalisaten in Forschungseinrichtungen, ergänzt durch Einsichtnahme der Bestände vor Ort. Unabdingbar war hierfür die Infrastruktur der Abteilung für Sondersammlungen der ULB Tirol (Arbeitsplätze, PCs und Laptops, gedruckte und elektronische Hilfsmittel, UV-Lampe, Leuchtfolie zur Analyse der Wasserzeichen usw.), aber auch der Bibliothek des Priesterseminars in der PhilosophischTheologischen Hochschule Brixen. 5.2 Zur Anlage der Handschriftenbeschreibungen Die Beschreibungen halten sich grundsätzlich an die „Richtlinien und Terminologie für die Handschriften beschreibung in Österreich“ der Kommission für Schrift- und Buchwesen (seit 2012 Institut für Mittel alterforschung, Abteilung Schrift- und Buchwesen) der ÖAW in Wien.195 Durch praktische Gepflogen heiten und technische Neuerungen notwendig gewordene Zusatzbestimmungen, die im Zuge des Austausches mit anderen HandschriftenbearbeiterInnen besprochen bzw. schriftlich niedergelegt wurden196, fanden auch in dieser Arbeit ihren Niederschlag. Bei einzelnen Fragestellungen waren die Kataloge der Handschriften der ULB Tirol sowie der Katalog der mittelalterlichen Handschriften der Bibliothek des Prämonstratenser Chorherrenstiftes Wilten in Innsbruck richtungsweisend.197 Zudem wurde den oben dargelegten methodischen Überlegungen Rechnung getragen und die Richtlinien dahin gehend adaptiert (siehe unten). Der Bibliothekssignatur als Kopfzeile folgt der Autor bzw. der anonyme Sachtitel, wobei bei umfangreicheren Textsammlungen ein fingierter Gesamttitel verwendet wird. Die Schlagzeile enthält relevante Daten zu Material, Umfang (gezählte Blätter), Maße des Buchblocks, Entstehungsort und Datierung der Handschrift. Die Beschreibung des Äußeren gliedert sich in mehrere Bereiche, gekennzeichnet durch folgende Siglen: B S N A E G L Beschreibstoff, Zustand und Zusammensetzung der Handschrift Schrift Notation Ausstattung Einband Geschichte der Handschrift Literatur Die Beschreibung des Äußeren bezieht sich in der Regel auf die Handschrift als Ganzes. Aufgrund der heterogenen Teile bei Sammelhandschriften wird dementsprechend eine allgemeine, die gesamte 195 196 197 O. Mazal, Richtlinien und Terminologie für die Handschriftenbeschreibung, in: Ders. (Hrsg.), Handschriftenbeschreibung in Österreich. Referate, Beratungen und Ergebnisse der Arbeitstagungen in Kremsmünster (1973) und Zwettl (1974) (Denkschriften der ÖAW, phil.-hist. Kl. 122 = Veröffentlichungen der Kommission für Schrift- und Buchwesen des Mittelalters II, 1). Wien 1975, 135–172. Vgl. G. Hayer, Richtlinien oder Dogma? Ein Beitrag zur Diskussion über die Richtlinien für die Handschriftenbeschreibung in Österreich, in: I. Reiffenstein (Hrsg.), Beiträge zur Überlieferung und Beschreibung deutscher Texte des Mittelalters. Referate der 8. Arbeitstagung österreichischer HandschriftenBearbeiter vom 25.–28.11.1981 in Rief bei Salzburg (Göppinger Arbeiten zur Germanistik 402). Göppingen 1983, 213–227; A. Derolez, Die Handschriftenbeschreibung und die Bibliotheks- und Buchgeschichte des Mittelalters, in: A. Rapp, M. Embach (Hrsg.), Rekonstruktion und Erschließung mittelalterlicher Bibliotheken (Beiträge zu den historischen Kulturwissenschaften 1). Berlin 2008, 105–118, hier 116–117. Wie Anm. 100; G. Kompatscher Gufler, D. Mairhofer, C. Schretter, Katalog der mittelalterlichen Handschriften der Bibliothek des Prämonstratenser Chorherren Stiftes Wilten. Redigiert von P. Ausserlechner (Denkschriften der ÖAW, phil.-hist. Kl. 425 = Veröffentlichungen der Kommission für Schrift- und Buchwesen des Mittelalters II, 10). Wien 2012. Zur Anlage des Katalogs 59 Handschrift betreffende Beschreibung des Äußeren an den Anfang gestellt; die einzelnen Teile werden mit speziellen Beschreibungen versehen. Abschnitt B verzeichnet Angaben zu Beschreibstoff, Erhaltungszustand, Zusammensetzung der Handschrift nach der Chroust’schen Lagenformel, Eigentümlichkeiten des Buchblocks, Existenz von Kustoden bzw. Reklamanten, Foliierung bzw. Paginierung sowie Textverlust. Die Angabe zu den Wasser zeichen reduziert sich auf einen Verweis auf die stetig erweiterte Internetplattform www.wzma.at. Verwandte Wasserzeichen werden nur im begründeten Anlassfall angeführt. Abreibungen sowie Betaradiografie-Aufnahmen der nachgewiesenen Wasserzeichen sind neben www.wzma.at auch über www.manuscripta.at ansteuerbar. Im Abschnitt S werden Informationen zur Schrift festgehalten: Schriftraum, Spaltenzahl, Zeilen zahl, Linierung, Schriftart, Schreiber, Schriftwechsel, Datierung. Da es sich bei den behandelten Hand schriften um einen Teilbestand handelt, konnten im Zuge detaillierter Untersuchungen des Komple mentärbestandes an der ULB Tirol Querverbindungen zu anderen Handschriften hergestellt werden. Die noch ausständige Untersuchung auch des archivalischen Materials in der Stiftsbibliothek bzw. im Stiftsarchiv in Neustift lässt weitere Erkenntnisse zu den Schreibern erwarten. Ebenso können Spuren zum Skriptorium des Augustiner Chorherrenstiftes Klosterneuburg noch weiter nachverfolgt werden. Hierfür soll dieser Katalog als Basis dienen. Die paläografische Terminologie folgt – mit leichten Adap tierungen – den Richtlinien Mazals.198 Die bei den vorliegenden Beschreibungen erstmals angeführte Sigle N bezieht sich auf Informa tionen zur Notation: Gestaltung, Notensysteme, Textzeilen, Art der Notation, Datierung und Lokalisierung, Besonderheiten. Die ausführlichen Beschreibungen der liturgischen Handschriften richten sich nach dem Vorbild der Innsbrucker Katalogbände. Abweichend davon werden die in den Missalien und Brevieren enthaltenen Sequenzen und Hymnen sowie die in den Sanktoralteilen ent haltenen Offizien in eigenen Tabellen übersichtlich dargestellt. Die Sigle A enthält Angaben zur Ausstattung der Handschrift. Aufgenommen wird sowohl einfa cher Dekor als auch höherrangiger Buchschmuck (s. Einleitung 3.2 Illuminierte Handschriften). Im Abschnitt E folgt eine Beschreibung des Einbandes. Angeführt werden Hinweise auf Material des Deckels und des Bezuges, Stil, Technik der Verzierung, Entstehungsort und Entstehungszeit. Auf grund der meist vorgenommenen Ablösung der ursprünglichen Einbände und Neubindung in Form von einfachen Halbfranz-Bibliothekseinbänden ist es leider nicht mehr möglich, eine umfassende Aus wertung der ursprünglichen Einbandtypen zu geben. Trotzdem wurde versucht, Querverbindungen zwischen den wenigen noch erhaltenen ursprünglichen Einbänden in Neustift und in anderen Tiroler Bibliotheken herzustellen und daraus buch- und bibliothekshistorisch relevante Schlüsse zu ziehen. Abbildungen der Einbände bzw. Abreibungen aller Einbandstempel und -rollen finden sich unter www.manuscripta.at bzw. im Abbildungsteil im Anhang. Abschnitt G fasst Hinweise auf die Geschichte der Handschrift zusammen und nennt alle Angaben über Vorbesitzer sowie Benützung der Handschrift. Damit soll versucht werden, die Handschrift in ihren bibliotheksgeschichtlichen Kontext zu betten und Zusammenhänge herzustellen. Um die histori schen Fakten deutlich darzustellen, werden alle entsprechenden Angaben im Wortlaut in möglichst chronologischer Reihenfolge wiedergegeben. Berücksichtigt werden u. a. Besitzeinträge (soweit zu ermitteln, auch getilgte), Kauf-, Schenkungsvermerke, Supralibros sowie Exlibris und alte Signaturen. Unter der Sigle L wird die gesamte Literatur, in der die Handschrift bzw. einzelne Teile daraus behandelt oder zumindest erwähnt bzw. zitiert sind, chronologisch geordnet aufgelistet. Auch Editionen, 198 Zur Frage der Nomenklatur vgl. K. Schneider, Paläographie und Handschriftenkunde für Germanisten. Eine Einführung. Tübingen 1999, 28 Anm. 46; Dies., Gotische Schriften in deutscher Sprache 1, 12–13; A. Fingernagel, Die Buchschriften des 12. Jahrhunderts, in: Ders. (Hrsg.), Romanik (Geschichte der Buchkultur 4/1). Graz 2007, 89–90; A. Derolez, The Palaeography of Gothic Manuscript Books. Cambridge 2003, 57; H. Foerster, Abriss der lateinischen Paläographie. Erweiterte 3. Auflage von T. Frenz. Stuttgart 2004, 217; J. Autenrieth, Probleme der Lokalisierung und Datierung von spätkarolingischen Schriften (10. und 11. Jahrhundert). Codicologia 4 (1978) 68. 60 Zur Anlage des Katalogs denen die betreffende Handschrift zugrunde liegt oder in deren Handschriftenlisten sie genannt ist, finden Berücksichtigung. Bibliografische Vollständigkeit wurde angestrebt, kann jedoch nicht zu gesichert werden. Mehrfach genannte Werke werden in Kurzform wiedergegeben und sind im Verzeichnis der abgekürzt zitierten Literatur aufgelistet. Relevante Publikationen ohne Nennung der Handschrift sind an entsprechender Stelle der Inhaltsbeschreibung angegeben. Im Anschluss an die Beschreibung des Äußeren folgt die Beschreibung des Inhalts. Einzelne Texte werden mit arabischen Ziffern durchnummeriert. In runden Klammern wird der Standort angegeben. Der Autor wird, wenn möglich, in normierter Form angeführt. Ist er in der Handschrift selbst nicht genannt, wird er in Spitzklammern gesetzt. Der Anführung des Titels folgen in Klammern der Nach weis des Textes in Standardeditionen oder zumindest in einem älteren Druck, Nachweise in gängigen Repertorien, Werks- bzw. Handschriftenverzeichnissen. Titel, Incipit und Explicit werden in der Regel nur dann angegeben, wenn sie im Wortlaut stark von den genannten Ausgaben abweichen oder keine gängige Edition vorliegt. Auch Schreibervermerke und Kleintexte werden berücksichtigt. Überschriften und Schlussschriften werden nur vermerkt, sofern sie nähere Angaben zum Text, z. B. die Zuschreibung zu einem anderen als dem ermittelten Autor, beinhalten. Die wiedergegebenen Textstellen werden im Wortlaut der Handschrift kursiv aufgenommen; im Initienregister finden sie sich in normalisierter Form. Eindeutige Abkürzungen werden ohne eigenen Ausweis aufgelöst. Auf etwaige Glossen ist hin gewiesen. Die noch in situ befindlichen Fragmente werden sowohl äußerlich als auch inhaltlich beschrieben. Zusammengehörige Fragmente in einer Handschrift werden an der Stelle ihres Erst vorkommens zusammengefasst. Von einer Beschreibung der abgelösten, (noch) nicht zugewiesenen Fragmente wurde abgesehen.199 Bisweilen folgen der inhaltlichen Beschreibung Hinweise zum Text, zur Textgeschichte, auf weiterführende Literatur oder zu Parallelüberlieferungen, vorwiegend im süd deutschen Raum. Kataloge mit Beschreibungen der zum Vergleich herangezogenen Handschriften werden nicht eigens ausgewiesen. Auch EDV-gestützte Hilfsmittel und Internetdatenbanken werden nicht eigens zitiert. Die Handhabung der Beschreibungen soll durch mehrere Register, Verzeichnisse und Übersichten erleichtert werden. Hinweise, Ergänzungen und Korrekturen zu den vorliegenden Beschreibungen sowie biblio grafische Addenda werden im Webportal www.manuscripta.at berücksichtigt. 199 Die Fragmente werden derzeit im Rahmen anderer Projekte erschlossen, vgl. Anm. 109. TEIL II: KATALOGTEIL 63 Verzeichnis der beschriebenen Handschriften Cod. 4 IOHANNES CALDERINUS. ARNOLDUS DE GEILHOVEN Pap. II, 432 Bl. 410 × 285. Oberitalien (Bologna?), Anfang 15. Jh. Cod. 7 GUILELMUS DURANTIS. IOHANNES ANDREAE Perg. I, 164, I Bl. 410 × 275. Bologna, 1. H. 14. Jh. Cod. 19 PETRUS LOMBARDUS Perg. I, 108, I* Bl. 385 × 270. Nordfrankreich (Raum Paris), 13. Jh. Cod. 20 VETUS TESTAMENTUM Perg. I, 168, I* Bl. 380 × 280. Entstehungsort unbestimmt, 12./13. Jh. Cod. 31 ALDOBRANDINUS DE TOSCANELLA. EXPOSITIO SUPER AVE MARIA. GODEFRIDUS HERILACENSIS Perg. I, 130, I* Bl. 320 × 240. Süddeutscher Raum (?), 14. Jh. Cod. 38 THOMAS DE AQUINO Perg. I, 272, I* Bl. 345 × 260. Süddeutscher Raum (?), 15. Jh. Cod. 52 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 273 Bl. 290 × 205. Süddeutscher Raum (Tirol?), T. I: Ende 14. Jh., T. II: 1396. Cod. 53 THOMAS EBENDORFER Pap. I, 348 Bl. 285 × 210. Wiener Raum (?), Mitte 15. Jh. (u. a. 1452). Cod. 76 SAMMELHANDSCHRIFT Perg./Pap. I, 75, I* Bl. 270 × 205. T. I: Neustift, Mitte 15. Jh., T. II: St. Dorothea (Wien) und Neustift, 6. Jz. 15. Jh. Cod. 77 THOMAS DE AQUINO. PETRUS BLESENSIS Pap. I, 120, I* Bl. 275 × 200. Basel, 1465. Cod. 78 PEREGRINUS OPPOLIENSIS Pap. I, 120, I* Bl. 280 × 205. Raum Brixen (Neustift?), um 1430/33. Cod. 83 GREGORIUS MAGNUS Pap. I, 59, I* Bl. 285 × 205. Süddeutscher Raum, 1400. 64 Verzeichnis der beschriebenen Handschriften Cod. 85 TRACTATUS DE SACRAMENTIS Pap. I, 107, I* Bl. 290 × 205. Wohl Neustift, Anfang 16. Jh. Cod. 86 THOMAS DE AQUINO Perg. I, 40, I* Bl. 300 × 205. Frankreich (?), wohl Mitte 13. Jh. Cod. 99 CLEMENS V. IOHANNES ANDREAE. IOHANNES XXII. Perg. I, 33, I* Bl. 400 × 270. Bologna, 1. H. 14. Jh. Cod. 100 MISSALE (FESTIVUM) Perg. 104 Bl. 380 × 295. Neustift, 1524–1526. Cod. 106 IOHANNES IANUENSIS Perg. I, 110, I* Bl. 365 × 285. Süddeutscher Raum (?), 1. H. 15. Jh. Cod. 107 COMPILATIONES ANTIQUAE I–III Perg. I, 81, I* Bl. 355 × 230. Nordfrankreich (Raum Paris), zw. 1210 und 1234. Cod. 123 BERNARDUS DE GORDONIO. MEDIZINISCHE TRAKTATE Pap. IV, 227, I* Bl. 315 × 215. Süddeutscher Raum, 6. Jz. 15. Jh. Cod. 125 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. I, 313, I* Bl. 310 × 220. Süddeutscher Raum, 6. Jz. 15. Jh. Cod. 142 REGULA S. AUGUSTINI. HUGO DE SANCTO VICTORE (?). LIBER ANNIVERSARIORUM NEOCELLENSIS. NECROLOGIUM NEOCELLENSE Perg. I, 173, I* Bl. 300 × 230. Neustift, T. I: 1521, T. II: 1459. Cod. 145 BARTHOLOMAEUS A SANCTO CONCORDIO Pap. I, 314, I* Bl. 300 × 215. Süddeutscher Raum (Tirol?), 1427. Cod. 146 REGEL DES HL. AUGUSTINUS. HUGO DE SANCTO VICTORE (?). HUMBERTUS DE ROMANIS Perg. I, 160, I* Bl. 290 × 220. Süddeutscher Raum (T. I: Neustift?), 1. H. 15. Jh. (vor 1445). Cod. 153 IOHANNES HEROLT (DISCIPULUS) Pap. I, 168, I* Bl. 270 × 205. Wiener Raum, 4. Jz. 15. Jh. Cod. 154 MISSALE SPECIALE (TEILMISSALE) Perg. I, 47, I* Bl. 295 × 195. Umfeld Neustift, 12. Jh. (mit Nachträgen 13. Jh.). Cod. 155 SAMMELHANDSCHRIFT Pap. I, 164, I* Bl. 290 × 205. Umfeld Neustift (u. a. Pfalzen), 7. Jz. 15. Jh. (u. a. 1463). Cod. 156 HUGO DE SANCTO CARO Perg. I, 88, I* Bl. 290 × 210. Nordfrankreich (Raum Paris), um 1300. Cod. 158 AUGUSTINUS Pap. III, 69, I* Bl. 285 × 205. Süddeutscher Raum, um 1400. Verzeichnis der beschriebenen Handschriften Cod. 160 LAURENTIUS RUSIUS Pap. I, 55, I* Bl. 290 × 210. Süddeutscher Raum, Mitte 15. Jh. Cod. 161 IOHANNES DE DAMBACH. ORIGENES. EXPOSITIO ORATIONIS DOMINICAE Pap. I, 12, I* Bl. 285 × 205. Süddeutscher Raum, Ende 14. Jh. Cod. 162 SERMONES IN DEDICATIONE ECCLESIAE Pap. I, 10, I* Bl. 290 × 205. Süddeutscher Raum, Ende 14. Jh. Cod. 163 ARISTOTELES Pap. I, 79, I* Bl. 290 × 220. Wien, T. I: um 1389 und um 1443, T. II: Ende 14. Jh. Cod. 165 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. I, 148, I* Bl. 295 × 205. Süddeutscher Raum, 1. Jz. 15. Jh. (T. III: 1407). Cod. 166 SPECULUM HUMANAE SALVATIONIS Pap. II, 58, I* Bl. 295 × 210. Süddeutscher Raum, 1. Jz. 15. Jh. Cod. 167 ADAM DE ALDERSBACH Pap. I, 72, I* Bl. 290 × 205. Süddeutscher Raum, 1419. Cod. 168 ARISTOTELES Pap. I, 46, I* Bl. 290 × 220. Wien, 9. Jz. 14. Jh. Cod. 171 THOMAS CANTIMPRATENSIS. THEOLOGISCHE TRAKTATE Perg. I, 91, I* Bl. 285 × 210. Süddeutscher Raum, Ende 13./Anfang 14. Jh. Cod. 173 MAURUS SERVIUS HONORATUS Pap./Perg. I, 172, I* Bl. 260 × 215. Italien, 2. Drittel 15. Jh. Cod. 174 SERMONES DE TEMPORE Pap. I, 225, I* Bl. 280 × 200. Tirol (?), um 1470 (u. a. 1472). Cod. 177 FLORES DE SANCTIS ET DE TEMPORE Pap. I, 153, I* Bl. 280 × 210. Tirol (Neustift?), um 1430/33. Cod. 181 ALANUS AB INSULIS Pap. I, 60, I* Bl. 285 × 210. Wiener Raum, um 1410. Cod. 182 PS.-EUSEBIUS Pap. I, 16, I* Bl. 285 × 205. Süddeutscher Raum, 1400. Cod. 183 HENRICUS SUSO Pap. I, 48, I* Bl. 285 × 205. Süddeutscher Raum, um 1400. Cod. 190 BRUNO DE SEGNI Perg. I, 68, I* Bl. 315 × 250. Neustift (?), 2. H. 15./Anfang 16. Jh. Cod. 194 RITUALE NEOCELLENSE Perg. I, 67 Bl. 305 × 225. Neustift, 1507. 65 66 Verzeichnis der beschriebenen Handschriften Cod. 196 LECTIONARIUM MISSAE NEOCELLENSE Perg. I, 26 Bl. 315 × 250. Neustift, Ende 15. Jh. (vor 1482?). Cod. 200 ROBERTUS HOLCOT Pap. I, 390, I* Bl. 310 × 215. Wiener Raum (?), um 1415/20. Cod. 204 VETUS TESTAMENTUM Pap. 307 Bl. 310 × 200. Tirol (?), um 1470. Cod. 217 DECIMUS IUNIUS IUVENALIS. AULUS PERSIUS FLACCUS Perg. I, 56, I* Bl. 310 × 200. Bologna, 1. H. 15. Jh. Cod. 252 ORDINARIUM MISSAE Perg. 46 Bl. 340 × 245. Neustift (?), letztes Viertel 15. Jh. Cod. 253 GODEFRIDUS DE TRANO. TANCREDUS BONONIENSIS. RAIMUNDUS DE PENNAFORTE Perg. 88 Bl. 380 × 265. Oberitalien (Bologna?), 1. H. 14. Jh. Cod. 282 BERNARDUS CLARAEVALLENSIS Perg. I, 25, I* Bl. 240 × 165. Entstehungsort unbestimmt, 2. H. 12. Jh. Cod. 293 VITA BEATI HARTMANNI EPISCOPI (BRIXINENSIS) Perg. 23, II* Bl. 260 × 170. Neustift, 12./13. Jh. Cod. 308 THEOLOGISCHE SAMMELHANDSCHRIFT Perg. I, 228, I* Bl. 220 × 150. Süddeutsch-österreichischer Raum, 14. Jh. Cod. 315 ISAIAS MIT GLOSSA ORDINARIA Perg. I, 88, I* Bl. 235 × 170. Entstehungsort unbestimmt, 12. Jh. Cod. 327 GALFREDUS DE VINOSALVO Perg. I, 21, I* Bl. 220 × 150. Frankreich (?), 13. Jh. Cod. 353 LIBER HORARUM, Bd. 1 (zusammengehörend mit Cod. 654) Perg. VI, 194 Bl. 160 × 120. Raum Brixen, um 1496. Cod. 360 CONRADUS HIRSAUGIENSIS Perg. I, 26, I* Bl. 165 × 110. Entstehungsort unbestimmt, 12. Jh. Cod. 397 IOHANNES DE TURRECREMATA Perg. I, 88, I* Bl. 140 × 105. Süddeutscher Raum (?), 2. H. 15. Jh. Cod. 405 BREVIARIUM SALISBURGENSE Perg. VII, 236 Bl. 135 × 100. Wohl Neustift, um 1500 (u. a. 1501). Cod. 440 MARSILIUS DE INGHEN. IOHANNES BURIDANUS Pap. I, 414 Bl. 280 × 205. Tirol (?), um 1450. Cod. 447 KANONISTISCHE SAMMELHANDSCHRIFT, u. a. ALBERTUS DE BRIXIA Pap. 208, X* Bl. 295 × 215. Süddeutscher Raum, 1414. Verzeichnis der beschriebenen Handschriften 67 Cod. 453 MEDIZINISCHE SAMMELHANDSCHRIFT Pap. III, 234, I* Bl. 295 × 210. T. I: Wiener Raum, um 1455–1460, T. II–IV: Wiener Raum (?), um 1440. Cod. 483 HONORIUS AUGUSTODUNENSIS Perg. I, 76, I* Bl. 255 × 170. Entstehungsort unbestimmt, 2. H. 12. Jh. Cod. 485 HUGO DE SANCTO VICTORE Perg. I, 66, I* Bl. 240 × 180. Entstehungsort unbestimmt, um 1200. Cod. 495 GREGORIUS MAGNUS Perg. 202 Bl. 210 × 140/145. Entstehungsort unbestimmt, 10. Jh. Cod. 532 SAMMELHANDSCHRIFT Perg. I, 145, I* Bl. 150 × 110. Entstehungsort unbestimmt, 14. Jh. Cod. 550 GUIDO DE MONTE ROCHERII. GUILELMUS DURANTIS Pap. I, 446, III* Bl. 295 × 210. T. I: Wiener Raum, 6. Jz. 15. Jh., T. II: Raum Brixen (u. a. Villanders), 7. Jz. 15. Jh. (u. a. 1465). Cod. 578 RAPHAEL VOLGOSIUS Pap. I, 10, I* Bl. 295 × 210. Brixen, 1480. Cod. 579 PUBLIUS OVIDIUS NASO Pap. 14 Bl. 300 × 210. Wiener Raum oder Tirol (?), 1491. Cod. 597 SERMONES. AUGUSTINUS FAVARONI Pap. 58 Bl. 270 × 210. Wiener Raum, um 1464/65. Cod. 605 PETRUS HISPANUS. DE MODIS SIGNIFICANDI Perg. I, 8, I* Bl. 185 × 135. Entstehungsort unbestimmt, Ende 13./Anfang 14. Jh. Cod. 606 TRACTATUS DE CONFESSIONE Perg. I, 6, I* Bl. 220 × 155. Entstehungsort unbestimmt, 14. Jh. Cod. 614 JURISTISCHE SAMMELHANDSCHRIFT Pap. I, 290, I* Bl. 210 × 145. Tirol, T. I: 7./8. Jz. 14. Jh, T. II: 1. Viertel 15. Jh. (u. a. 1419), T. III: 1. Jz. 15. Jh., T. IV: 4. Jz. 15. Jh. Cod. 617 SAMMELHANDSCHRIFT Pap. I, 282 Bl. 220 × 145. Tirol (u. a. Neustift), T. I, II, IV: 7. Jz. 15. Jh. (u. a. 1467), T. III: 6. Jz. 15. Jh. (u. a. 1455). Cod. 654 LIBER HORARUM, Bd. 2 (zusammengehörend mit Cod. 353) Perg. II, 207, I* Bl. 165 × 120. Raum Brixen, 1496. Cod. 691 ARGUMENTA LIBRORUM DICTYS CRETENSIS Pap. 5 Bl. 210 × 145. Neustift (?), Anfang 16. Jh. Cod. 694 PAULUS URSINUS Pap. 27 Bl. 205 × 150. Tirol, 1523. 68 Verzeichnis der beschriebenen Handschriften Cod. 726 RITUALE NEOCELLENSE Pap. III, 78, I* Bl. 120 × 100. Neustift, 1523 (mit Zusätzen 16. und 17. Jh.). Cod. 748 MEDIZINISCHE SAMMELHANDSCHRIFT Pap. III, 220 Bl. 290 × 215. T. I–III: Wiener Raum (?), um 1440, T. IV: Schwäbisch-Schweizer Raum, um 1430. Cod. 754 THEOLOGISCHE SAMMELHANDSCHRIFT, u. a. IOHANNES GERSON Pap. 26 Bl. 280 × 195. Tirol, 3. Viertel 15. Jh. Cod. 755 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 12 Bl. 270 × 200. Neustift oder Wiener Raum (?), 6. Jz. 15. Jh., 1481 (?) und 16. Jh. Cod. 756 MATTHIAS ENGELSCHALK Pap. 53 Bl. 275 × 200. Süddeutscher Raum, 1479. Cod. 776 MUSIKALISCH-ASTRONOMISCHE SAMMELHANDSCHRIFT Pap. I, 71, I* Bl. 210 × 155. Neustift (?), Anfang 16. Jh. (u. a. 1502). Cod. 780 GAIUS SALLUSTIUS CRISPUS Pap. 86 Bl. 210 × 145. Oberitalien (?), Ende 15. Jh. Sign. 14820 RITUALE NEOCELLENSE Perg. X, 85, II* Bl. 300 × 230. Neustift, Anfang 16. Jh. (mit Zusätzen, u. a. 1578). Sign. 15063 ANTIPHONARIUM Perg. 197 Bl. 355 × 255. Wohl Neustift, 2. H. 15. Jh. (mit späteren Zusätzen). Nr. 139 (olim Sign. 14819) GRADUALE. ANTIPHONARIUM Pap. 231 Bl. 315 × 215. Umfeld Neustift, Ende 15. Jh. (T. I: 1495/96). Nr. 940 GRADUALE. ANTIPHONARIUM Pap. I, 229, I* Bl. 310 × 220. Umfeld Neustift, 1490/93. ohne Sign. NEUSTIFTER WAPPENBUCH Perg. I, 88, I* Bl. 245 × 180. Sterzing (?), 1548–1552. ohne Sign. GRADUALE NEOCELLENSE I Perg. II, 289, I* Bl. 725 × 505. Neustift, 1442. ohne Sign. GRADUALE NEOCELLENSE II Perg. I, 317, I* Bl. 700 × 500. Neustift, Mitte 15. Jh. (zw. 1442 und 1446). 69 Verzeichnis der Abkürzungen und Siglen Vorbemerkung: Abkürzungen von biblischen Büchern nach der Biblia sacra iuxta vulgatam versionem (Ed. R. Weber, B. Fischer. Stuttgartiae 331983, XXXI), Abkürzungen von Festbezeichnungen weitgehend nach Schneyer (s. Literaturverzeichnis), Abkürzungen als Teile von Handschriftensignaturen meist ohne Auflösung. A Abb. Abh. Abt. Anm. ap. App. art. B Bd., Bde bearb. bes. Bibl. Bl. BMV BN BSB bzw. C. ca. cap. CC CCl Clm Cod. d. h. dat. ders., dies. Dipl.-Arb. Diss. dist. dnca dni Doppelbl. dt. E ebda Ed. Ausstattung der Handschrift Abbildung Abhandlung(en) Abteilung Anmerkung apostolus, apostoli Appendix articulus Beschreibstoff und Zusammensetzung der Handschrift Band, Bände bearbeitet besonders Bibliothek, Bibliotheca, Biblioteca, Bibliothèque Blatt, Blätter Beata Maria Virgo Bibliothèque Nationale Bayerische Staatsbibliothek beziehungsweise Causa circa capitulum, Kapitel Codex Cremifanensis Codex Claustroneoburgensis Codex Latinus Monacensis Codex, Codices das heißt datiert derselbe, dieselbe Diplomarbeit Dissertation distinctio dominica domini Doppelblatt, -blätter deutsch Einband ebenda Editio(n) ed. ehem. Einzelbl. ep. etc. ev. evtl. Expl. fasc. fer. ff. Forts. fr. Fragm. G Gegenbl. gest. H. HD hl. hrsg. Hrsg. Hs. i. m. Inc. Ink. insbes. Jh. Jz. korr. L lat. LB lib. Lit. mm Ms. mut. N Nachsatzbl. edidit, ediderunt, editio ehemals, ehemalig Einzelblatt, -blätter epistola et cetera evangelista eventuell Explicit fasciculus feria (und) folgende Fortsetzung frater Fragment Geschichte Gegenblatt, -blätter gestorben Hälfte Hinterdeckel heilig(e) herausgegeben Herausgeber Handschrift, Handschriften in margine Incipit Inkunabel insbesondere Jahrhundert Jahrzehnt korrigiert Literatur lateinisch Landesbibliothek liber, libri Literatur Millimeter Manuskript (Titel, Incipit, Explicit) mutile Notation Nachsatzblatt, -blätter 70 ND N. F. Nr. o. ÖAW ÖNB Pap. Perg. phil.-hist. Kl. Ps.qu. Quadrages. quaestiunc. Quinquages. r S S. s. s. s. d. s. o. s. u. s. v. SB Septuages. Ser. Sexages. Sign. sog. Verzeichnis der Abkürzungen und Siglen Nachdruck Neue Folge Nummer ohne Österreichische Akademie der Wissenschaften Österreichische Nationalbibliothek Papier Pergament philosophisch-historische Klasse Pseudoquaestio Quadragesima quaestiuncula Quinquagesima recto Schrift Seite sanctus siehe siehe dort siehe oben siehe unten sub verbo Sitzungsberichte Septuagesima Serie, series Sexagesima Signatur sogenannt Auflösung der Siglen der Bearbeiterinnen und Bearbeiter: A. P.: Anna Pinter C. S.: Claudia Schretter-Picker G. G.: Giulia Gabrielli G. K.: Gabriela Kompatscher Gufler L. Š.: Lav Šubarić M. S.: Maria Stieglecker P. A.: Petra Ausserlechner P. K.: Patrik Kennel S. R.: Susanne Rischpler U. S.: Ursula Stampfer W. N.: Walter Neuhauser Sp. ss. Staatsbibl. Stadtbibl. Stiftsbibl. Str. Suppl. T. Taf. Tit. tit. tract. tw. u. a. u. ö. UB ULB urspr. V. v v. a. Vaticana VD vgl. Vorsatzbl. Wiss. Stadtbibl. Z. z. B. z. T. zw. Spalte sanctissimus Staatsbibliothek Stadtbibliothek Stiftsbibliothek Strophe Supplement Teil Tafel Titel titulus tractatus teilweise und andere(s), unter anderem und öfter Universitätsbibliothek Universitäts- und Landesbibliothek ursprünglich Vers verso vor allem Bibliotheca Apostolica Vaticana Vorderdeckel vergleiche Vorsatzblatt, -blätter Wissenschaftliche Stadtbibliothek Zeile zum Beispiel zum Teil zwischen 71 Verzeichnis der abgekürzt zitierten Literatur 850 Jahre Augustiner Chorherrenstift Neustift 850 Jahre Augustiner Chorherrenstift Neustift, hrsg. vom Augustiner Chorherrenstift Neustift. Brixen 1992. 850 Jahre Neustift. Katalog Th. (H.) Innerhofer (Red.), 850 Jahre Chorherrenstift Neustift. 1. Südtiroler Landesausstellung. Stift Neustift 30. Mai bis 31. Oktober 1992. Katalog. Brixen 1992. AH Analecta hymnica medii aevi. Ed. G. M. Dreves, C. Blume. 55 Bde. Leipzig 1886– 1922, ND Frankfurt 1961. 2 Registerbde. Bern, München 1978. Atz K. 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Österreichische Akademie der Wissenschaften. www.wzma.at Sämtliche Weblinks wurden zuletzt im April 2021 überprüft. KATALOG Cod. 4 81 Cod. 4 IOHANNES CALDERINUS. ARNOLDUS DE GEILHOVEN Pap. II, 432 Bl. 410 × 285. Oberitalien (Bologna?), Anfang 15. Jh. B: Pap. leicht gewellt, vereinzelt kleine Löcher und Risse, tw. verschmutzt, vereinzelt Tintenkleckse, am oberen Blattrand Spuren eines Wasserschadens, insbes. am Beginn und am Schluss (mit Beschädigung der Malerei). Zw. Bl. 46 und 47 (am Falz) Spuren von gepressten pflanzlichen Blättern. Nachweise und Bilder der Wasser zeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: III (Vorsatzlage, Perg.) + 11.V110 + IV118 + 6.V178 + (IV+2)188 + 24.V428 + (III–2)432. Bl. I urspr. als Spiegel auf dem VD aufgeklebt, Bl. II stark beschnit ten (110/115 × 280). Bl. 179 und 188 eingehängte Einzelbl. Gegenbl. zu Bl. 429 und 430 herausgeschnitten (ohne Textverlust). Lagenreklamanten. Am unteren Blattrand mittig Kennzeichnung der Blätter der ersten Lagenhälfte mit Kleinbuchstaben in alphabetischer Reihenfolge (auf z folgt aa), vielfach stark be- bzw. weggeschnitten, am rechten unteren Blattrand vereinzelt Zählung der Blätter der ersten Lagenhälfte in arabischen Ziffern erhalten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 215 ausgelassen, 280 doppelt angeführt. Rote Ledersignakel. S: Schriftraum 260/270 × 180/185. Zwei Spalten, von einfachen Bleistift- bzw. Blindlinien gerahmt, zu 52–65 Zeilen. Bastarda: Rudolph Claghewinter de Saxonia (Bl. 431vb), vgl. Krämer, Scriptores. Am Blattrand Korrekturen, zahlreiche Glossen, meist von anderer Hand, vereinzelt gerahmt. Außerhalb des Schriftraumes Notazeichen. A: Abwechselnd rote und blaue Paragraphzeichen; diverse Verweiszeichen (oft drei Punkte mit kommaartigem Fadenausläufer); des Öfteren tintenfarbene Randmarkierungen, u. a. Bl. 62rb Doppellinie, aus der ein Drei eck (mit Bögchenrand) ragt, in das ein kleiner Kreis eingetragen ist; Lagenreklamanten von je vier Drei punktmotiven gerahmt; ab Bl. 17r zahlreiche flüchtige Zeigehände (mit Ärmelansatz); Bl. 17rb, 70va–b (jeweils am Kolumnenrand) Profilgesichter. In der Regel zwei- und dreizeilige Lombarden, abwechselnd in Rot und Blau; die blauen Lombarden mit rotem, die roten Lombarden mit violettem Fleuronnée. Repräsen tanten. Fleuronnée: in den Binnenfeldern und als Besatz charakteristische saitenartige Parallelstriche, besetzt mit kleinen Perlen oder henkelartigen Klammern; schwungvoll zurückgebogene Fadenausläufer, tw. lockere (bzw. ondulierende) Bündelfäden und florale Endmotive (kleine Häkchenblüten, Bl. 373vb „Ähre“). – Fleuronnée-Initialen jeweils zu Beginn der Lemmata, Paragraphzeichen jeweils zur Bezeichnung der Unter abschnitte eines Lemmas. Deckfarbenschmuck: 24 neun- bis 17-zeilige ornamentale Initialen mit kurzen Ranken (nur Bl. 1r mit dreiseitigem Rankenwerk) am Beginn der Praefatio und der einzelnen Buchstaben dieses alphabetisch gegliederten Repertoriums; Bl. 1r ein Wappen; Bl. 1r Freifläche (ca. 185 × 110) für einleitende Miniatur (Dedikationsbild?), nicht ausgeführt. Die Ornamentalinitialen (Bl. 1ra, 1va [spaltenbreit], 43ra, 49vb, 105vb, 134vb, 171va, 184vb, 187rb, 193va, 233vb, 234va, 251vb, 278va, 288vb, 299ra, 350rb, 353ra, 369rb, 398vb, 410vb, 430va, 431va, 431vb [bes. beschädigt]) mit blattvergoldeten, schwarz konturierten Außengründen. Buchstabenkörper häufig in abge stuften Rosa- und Lilatönen (auch Grau), gerne mit feinperligem „Erbsenschotenmotiv“ (dieses tw. auch an Blättern) als Randdekor, seltener über Kreuz geschlungene Bänder (z. B. Bl. 1ra) als Ziermotiv. In den Binnenfeldern zumeist Blattformationen auf blauem oder karminrosafarbenem Grund, dieser mit feinen, weißen, tw. spiralig eingerollten Blättchenranken mit kugeligen Endhäkchen; feiner weißer Lineardekor auch auf Buchstabenkörpern und Ranken. Die kraft- und schwungvoll gewundenen Ranken entwachsen den Buchstabenkörpern, drehen sich an den Enden fallweise ein und/oder laufen in tintenfarbene Fäden aus; diese zuweilen schwer von den immer wieder sichtbaren Vorzeichnungslinien zu unterscheiden (z. B. Bl. 184vb); selten bilden die Rankenäste spitze Haken aus (z. B. Bl. 369rb); Rankenblätter mit feiner Strichelung in dunklerer Lokalfarbe abschattiert; als Dekor dicht gestreute, meist freie, schwarz konturierte Goldpunkte mit relativ langem Stiel, der an kleinem Haken bzw. Punkt ansetzt; im Rankenwerk der 82 Cod. 4 inleitungsseite auch Goldfelder. Kräftiges, die Grundfarben Rot, Blau und Grün vielfältig nuancierendes E Kolorit; neben Blattgold (auf hellrosafarbenem Bolus) auch Pinselgold verwendet. Bl. 1r mittig auf dem Bas-de-page Wappen (in einem aus vier Bögen und vier Spitzen zusammengesetzten Vierpassrahmen; auf grünem, von schwarzen Blättchenranken überzogenem Grund): in Gold ein abgerissener schwarzer Greifenkopf, umgeben von drei achtstrahligen schwarzen Sternen; Wappen bislang uniden tifiziert. Deckfarbenschmuck wie im Falle von Brixen, Priesterseminarbibl., Cod. C.6 durch das Spätwerk des 1349 bis 1403 nachweisbaren Niccolò di Giacomo da Bologna beeinflusst (s. M. Roland, Fleuronnée als Binde glied. Das Brevier des Ludwig von Teck und die Fleuronnée-Gruppe St. Dorothea II, in: M. Haltrich, M. Stieglecker [Hrsg.], Code(x). Festgabe zum 65. Geburtstag von Alois Haidinger [Codices Manuscripti. Suppl. 2]. Purkersdorf 2010, 104–131, hier 109–110, insbes. Abb. 29), was zeitlich gut mit dem Wasser zeichenbefund zusammengeht. Bei Niccolò di Giacomo ähnlicher Aufbau von Einleitungsseiten (Wappen, in Medaillon, auf dem Bas-de-page in symmetrisches Rankengefüge eingegliedert), ebenfalls großzügig verwendete freie Goldpunkte mit schwarzer Kontur und Stiel sowie Verzicht auf Tierdrolerien. Das Fleuronnée ebenfalls italienisch. S. R. E: Schmuckloser gotischer Einband: weißes Schweinsleder über Holz. Tirol (?), 2. H. 15. Jh. Leder zerkratzt und fleckig, Holz tw. freiliegend. Am VD und HD Spuren von je fünf rautenförmigen Be schlägen. Am VD oben mittig Spuren eines Titelschildes. Am HD oben mittig Spur eines runden Beschlags, urspr. wohl Kettenbuch. Reste von zwei Leder-Metall-Schließen, verziert mit achtblättriger Rosette (wie Neustift, Stiftsbibl., Cod. 440). Kanten gerade, Unterkanten stark abgerieben. Rücken: vier Doppelbünde. Im ersten Feld aufgeklebtes Papierschild M: S: sowie Repertorium Iuris, im zweiten Signaturschild (s. bei G), im untersten Spuren eines urspr. aufgeklebten Signaturschildes (?). Kapitale mit naturfarbenem Spagat umstochen, oberes Kapital beschädigt. Urspr. Spiegel des VD Pergamentblatt, heute gelöst (s. bei B), auf überlappendem Leder des Einbandes Signaturen und Bleistiftnotiz, auf Holz Exlibris aufgeklebt (s. bei G), Spiegel des HD leeres Pergamentblatt. G: Bl. IIv Vermerk über die testamentarische Überlassung an das Kloster Neustift durch den Domherrn Johann Ebner (möglicherweise gebürtig aus Krems, seit 1442 als Brixner Domherr belegt, vgl. Santifaller, Brixner Domkapitel 298, Nr. 55) mit Hinweis auf seinen Tod am 3. August 1460 (laut Santifaller Todesdatum aller dings 3. August 1461): Iste liber legatus est monasterio sancte Marie virginis ad gracias alias Novacella communiter nuncupata prope Brixinam per venerabilem virum magistrum Iohannem Ebner canonicum Bri xinensem ob salutem anime sue. Qui obiit anno domini MoCCCCoLXo die 3a Augusti. Cuius anima requiescat in pace. Amen. Ähnlich lautende Vermerke Bl. 234r (Iste liber pertinet monasterio sancte Marie in Novacella prope Brixinam datus a magistro Iohanne Ebner Ao domini etc. 60 3a die Augusti) und Bl. 431va (Iste liber legatus est monasterio Novacellensi prope Brixinam per venerabilem virum quondam magistrum Iohannem Ebner canonicum Brixinensem pro remedio anime sue Ao 1460 die 3a Augusti). Bl. 1r Besitzvermerk 17. Jh. Collegii Novacellensis. Auf der Innenseite des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Auf der Innenseite des VD alte Signaturen der ULB Tirol II`1 B 3 (Tinte) und Nr. 4 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 4. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 136, II 108, 110. – Wretschko–Sprung 13. – Hermann 123, Nr. 144. – L. Santifaller, Das Brixner Domkapitel in seiner persönlichen Zusammensetzung im Mittelalter (Schlern-Schriften 7). Innsbruck 1924, 298, Nr. 55 (ohne Nennung der Sign.). – Neuhauser, Neustift 88. – Colophons Nr. 16308. – Katalog ULB Tirol I 59, 281. – Stefani 122–123. – Peintner, Stiftsbibliothek 122. – Krämer, Scriptores, s. v. „Claghewinter, Rudolphus“. (Ir) leer. (Iv) Federproben: Zwei Verse eines italienischen Strambotto, 15. Jh. (vgl. V. Cian, Ballate e strambotti del sec. XV. tratti da un codice Trevisano. Giornale storico della letteratura italiana 4 [1884] 1–55, hier 53). zamay non spiero de veder vendeta / de quella falsa lengua maledeta. (IIr) leer. (IIv) Vermerk über die testamentarische Überlassung (s. bei G). Bleistiftnotiz (19. Jh.): Kummt nächstens zur vollständigen behandlung. Cod. 7 83 (1ra–431vb) ‹Iohannes Calderinus›: Repertorium utriusque iuris (Druck: GW 5904), mit Praefatio von Arnoldus de Geilhoven: Remissorium utriusque iuris (CCCM 212, XII). (1r) Tit. (18. Jh.): Anno (gestrichen: 1460) Repertorium iuris scriptum. (1ra) Praefatio (tw. Druck: A. Rivier, Dr. Arnold Gheyloven, aus Rotterdam, Verfasser eines Remisso rium juris utriusque und anderer juristischer Schriften. ZRG Germanistische Abt. 11 [1873] 454–455). Inc.: Memoriam esse thesaurum inventorum eleganter ait Cycero in Rethoricis ad Herenium libro tercio … – Expl.: … fundere penes omnium conditorem. Qui trinus et unus vivit et regnat in secula benedictus Amen. (1va) Text. Inc.: A hec dictio. A Interdum includit ut C de … – Expl.: … Zenotasium quid nota ff de rerum divisione § intactum et cetera. Laus tibi Christe. Explicit. Die Verfasserfrage lässt sich für den Haupttext nicht eindeutig klären. Der Text wird in der Parallel überlieferung tw. Iohannes Calderinus, tw. Arnoldus de Geilhoven zugeschrieben. Das Incipit (Bl. 1va) stimmt mit Schulte GQ II 249–250 (Iohannes Calderinus, Repertorium utriusque iuris) überein. Bei F. Schulte, Die canonistischen Handschriften der Bibliotheken in Prag. Mit drei Tafeln. Abh. der böh mischen Gesellschaft der Wissenschaften VI/2 (1868) 77, Nr. 190 wird dasselbe Inc. Arnoldus de Geilhoven zugeschrieben. Lit.: Schulte GQ II 249–250 und 438–439. (414ra–b) leer bis auf Notiz: reverte folium scribitur (?): per Io. An. de fide instru. (431vb) Schreibervermerk: Scriptum per me Rodulphum (!) Claghewinter de Saxonia. (431v) Bleistiftnotiz (19. Jh.): Vide abietiv zu behandeln. (432r–v) leer. A. P. / U. S. Cod. 7 GUILELMUS DURANTIS. IOHANNES ANDREAE Perg. I, 164, I* Bl. 410 × 275. Bologna, 1. H. 14. Jh. B: Zwei Teile: I (I, 1–143[144]), II (144[145]–163[164], I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 52 doppelt angeführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. stark abgerieben, insbes. an den unteren Kanten. Ecken mit braunem Leder verstärkt. Rücken: braunesLeder, fünf einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck IUS CANONIC. GLOSAT., im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotizen (s. bei G), Spiegel des HD leer. G: Die Hs. dürfte wohl zu jenen vier Hs. kirchenrechtlichen Inhalts zählen, die 1353 als Geschenk des Herrn Berthold, Hospitaler zu Brixen, in den Besitz des Stiftes gelangten (vgl. Neustift, Stiftsarchiv, WW 59; Mairhofer 264–265; Peintner, Stiftsbibliothek 118; Peintner, Schreibkunst 377). Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`1 J 11 (?, radiert), II`2 A 7 (gestrichen), II 1 B 6 (Bleistift), Nr. 7 (Tinte) und Bleistiftnotizen (Vide Grundzettel) sowie (wird paläograf. behandelt und vollständig katalog.). Am Rücken gelbes Signaturschild der ULB Tirol 7. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat ab gegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 136. – Wretschko–Sprung 13. – Rasmo 20 (Nr. 13–14) und Abb. 11–13 (ohne Nennung der Sign.). – Furtenbach–Kalb 21. – Neuhauser, Neustift 88. – Peintner, Buchmalerei 33, 41 und 42. – Katalog ULB Tirol I 62, 281. – Peintner, Stiftsbibliothek 118. – M. Bollati, Kat.-Nr. 13: Dekretalien, in: A. De Marchi, 84 Cod. 7 T. Franco, S. Spada Pintarelli (Hrsg.), Trecento. Gotische Maler in Bozen. Ausstellungskatalog. Bozen 2000, 105–107. – Peintner, Schreibkunst 377. I (I, 1–143[144]) B: Haar- und Fleischseite deutlich unterscheidbar, tw. fleckig. Vereinzelt Nahtspuren. Bl. 141 zw. den zwei Spalten in der Blattmitte ein Stück Perg. herausgeschnitten (wohl ohne Buchschmuck). Am Blattrand leicht gewellt. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + (V–III)4 + 14.V143(144). Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Die äußersten drei Doppelbl. der ersten Lage verloren (Textverlust). Meist Lagen reklamanten, gerahmt und mit einfachen Ausläufern versehen. Zw. Bl. 63 und 64 dürfte mindestens eine Lage fehlen (Textverlust). Am oberen Blattrand Einteilung der Bücher (1–4), im ersten Buch mit zusätzlicher Angabe der Partes; bei den ersten zwei Büchern urspr. Einteilung (primi, secundi) durchgestrichen und durch Zahlen ersetzt. S: Schriftraum 325/345 × 185. Zwei Spalten, von Tintenlinien gerahmt, zu 87–93 Zeilen auf Tintenlinierung. Zirkelstiche. Textualis formata (Littera Bononiensis). Vereinzelt Korrekturen und Ergänzungen am Schrift rand tw. von gleicher, tw. von späterer Hand. Zahlreiche Orientierungshilfen in Vierpassform am Blattrand (s. bei A). A: Rote Überschriften, tw. nicht ausgeführt, vereinzelt von späterer Hand in Schwarz am Schriftrand ergänzt; die Vierpassrahmen (s. bei S) mit Fibrillen und gestrichelten Rändern (z. B. Bl. 5vb, 89va), von dieser Hand auch winzige Vorschreibungen auf den Seitenrändern (z. B. Bl. 134va) und Zeilenfüllsel (Bl. 32rb am Ko lumnenende verzierte „Kugeln“, durch Reihen von o- bzw. c-förmigen, vom Schreiber ausgeführten Glie dern zu einer Art Kette verbunden); abwechselnd rote und blaue Paragraphzeichen; in Tintenfarbe: Rand markierungen (z. B. Bl. 112va Vertikallinien im Wechsel mit Gruppen kleiner Perlen), Lagenreklamanten mit Rahmen und Fibrillen (z. B. Bl. 14v) sowie flüchtige Zeigehände (z. B. Bl. 25ra). Repräsentanten. Zu Beginn von Unterabschnitten zwei- bis fünf-, meist dreizeilige Lombarden, abwechselnd in Rot und Blau (zuweilen mit Tendenz zu Violett), umfasst und ergänzt von einem großen, gegenfarbigen Paragraphzeichen mit langem, an der Kolumne anliegendem Ausläufer; Lombarde und Paragraphzeichen jeweils in der Gegen farbe mit Fleuronnée besetzt, das wiederum in der Gegenfarbe gepunktet, gekernt und/oder gestrichelt sein kann; nur selten an einem der Bestandteile das Fleuronnée weggelassen (z. B. Bl. 45v). Charakteristisch, im Binnenfeld und als Besatz, das Motiv der Fadenspirale; in ihrem Zentrum fallweise eine (aus vollfarbigem Grund ausgesparte) Dreierformation aus Knospenkopf und zwei Perlen; häufig auch aus dem Binnenfeld grund ausgesparte, langstielige Knospen. Als Besatz Perlen: klein und mit Abstand aufgereiht oder größer und eng aneinandergesetzt (tw. gepunktet bzw. besetzt mit kleinen Perlen); häufig einfache, etwas unsicher geführte Fadenausläufer mit volutenartig eingerollten Enden. Bl. 29va, 29vb und 30rb der Fleuronnée- Besatz der Paragraphzeichen (hier mit Zickzack-Aussparungen) wohl von anderer Hand, s. v. a. die Faden ausläuferbündel. Bl. 5v–10v zumeist in Initialnähe von späterer Hand ergänzte Federzeichnungen (stark verblasst; abgerie ben?), häufig Figuren (z. B. Bl. 10v im Interkolumnium stehender Mönch?), die mit der Initiale interagieren können (z. B. Bl. 5va links unten sitzender Mann, der Ausläuferblätter berührt). Deckfarbenschmuck: zu Beginn der Kapitel in der Regel dreizeilige (auch zwei- bis fünfzeilige) figürliche Initialen, tw. mit Serifen- oder mittig am Initialfeld ansetzendem Dekorblatt; nur wenige kurze Ranken; bei I-Initialen ersetzen die Büsten (s. u.) den Buchstaben (z. B. Bl. 96vb1); im vierten Buch auch einige Orna mentalinitialen zu Kapitelbeginn (mit einfachem Dekor, z. B. Bl. 87va); zu Beginn des 2. und 4. Buches je eine spaltenbreite Miniatur (24–29 bzw. 22–25 Zeilen hoch), jeweils unmittelbar gefolgt von einer größeren Einleitungsinitiale (acht bzw. 13 Zeilen hoch, jeweils mit kürzerer Blattranke und zwei Goldpunkten); eine größere Initiale auch zu Beginn von De sponsalibus et matrimoniis (Bl. 141vb2, siebenzeilig, ein Ausläufer blatt). – Die Initialen mit blattvergoldeten, schwarz umrandeten, meist quadratischen Außengründen, Buch stabenkörper häufig rosa(rot), seltener grau(grün), mit feinem weißen Liniendekor, ebenso die blauen Bin nenfelder; die seltenen kurzen Ranken (nur zu Beginn des Werks) mit tintenfarbenen Fadenausläufern sowie konturierten (und gestielten) Goldpunkten. In den kleinen Binnenfeldern zumeist männliche Büsten oder Köpfe (Inkarnat-Abschattierung mit Grau), fallweise (aber nicht unbedingt mit Textbezug) als Kleriker ge kennzeichnet, beispielsweise tonsurierter Mönch (z. B. Bl. 75ra3) oder Bischof mit Mitra (z. B. Bl. 141vb2); bei I-Initialen neigen sich die Figuren zuweilen aus den Initialfeldern heraus (z. B. Bl. 79va: Figur präsen tiert Schriftstück, Beischrift libellus) oder entsteigen ihnen (z. B. Bl. 76rb4). Textbezug z. B. Bl. 131va (De donationibus): Büste eines Mannes, der (Geld-)Beutel emporhält (durchscheinende Konturlinie belegt, dass Haltearm nicht zur urspr. Konzeption gehörte). Bl. 32va Miniatur: Gerichtsszene zw. zwei turmartigen Gebäuden (mit Bauzier und Einblicken in Kuppel räume) mit zentral auf einem einfachen Steinsitz thronendem König als Richter, flankiert von je drei Cod. 7 85 Männern (u. a. Kleriker mit grauer Kutte und Gelehrter mit purpurroter Kappe), die Schriftstücke präsentie ren; König wendet sich (mit Redegestus?) dem grauhaarigen Mann zu seiner Rechten zu; zu seinen Füßen purpurrotes Fußpolster, hinter ihm goldene Wand (Blattgold tw. abgeplatzt, hellrosafarbener Bolus und Vorzeichnung sichtbar), über die hellblaues, reich gefälteltes Tuch herabhängt. – Die anschließende Initiale zeigt bärtigen Mann mit erhobener Rechter. Bl. 66ra Marienkrönung: Hinter dem breiten, reich verzierten Steinthron, auf dem Christus seine Mutter krönt, fünf Engel (mit spitzen Flügeln), die goldenes Ehrentuch entfalten; Thron flankiert von zwei stehen den Engeln mit goldenen Stäben; Christus hält weißen Herrscherstab in der Linken; goldfarbenes Fußpols ter; Initialfeldandeutung durch purpurnen, blau umrandeten Hintergrund in Spaltenbreite. – In zugehöriger Initiale Schöpfung Adams, der nackt, den Kopf auf seine Linke gestützt, auf schräg ansteigender Felsforma tion liegt; anatomische Unsicherheiten (Ansatz des linken Arms). Gott, der außerhalb des hochrechteckigen Initialfelds steht, neigt sich Adam zu. Bl. 141vb2 Büste eines graubärtigen Bischofs mit rotem Mantel über weißem Gewand und goldverzierter Mitra. M. Bollati hat den Deckfarbendekor des Cod. 7 dem „Maestro del B 18“ (alias „Secondo Maestro di San Domenico“) zugeschrieben, was für T. I zutrifft. Der „Maestro del B 18“ lässt sich in den ersten Jahrzehnten des 14. Jh. v. a. in Bologna nachweisen; zu diesem Illuminator s. auch S. L’Engle, Master of B18, the Roer mond Volumen Parvum and the Early Fourtheenth Century Illumination in Bologna. Codices Manuscripti 52/53 (2005) 1–20. – Das Fleuronnée in T. I ist oberitalienisch. S. R. 1 (Ir–v) leer. (1ra–143vb) ‹Guilelmus Durantis›: Speculum iudiciale, unvollständig (Druck: GW 9148–9161. Basel 1574 u. ö.). (1ra–32rb) Lib. 1. Inc. mut.: Item quod est frater …] paternam pietatem non agnoscat … Quid si aliquis interfecit patrem vel tutorem meum … (= Particula 2, cap. De accusatore) – Expl.: … Qualiter autem libelli super salariis concipiantur dicetur I. in T. de magistris. (1ra–2rb) Particula 2, beginnend in cap. De accusatore. (2rb–10va) Particula 3. Mit Fehlstelle zw. Bl. 4vb und 5ra (innerhalb cap. De procuratore). (4vb) Expl. mut.: … hoc fallit in contractu matrimonii Extra De conver. i[ – (5ra) Inc. mut.: ] quod non quia tantum consequitur per directam … (10va–32rb) Particula 4. (32va–63vb) Lib. 2. Inc.: Sancti spiritus gracia suffragante de personis … – Expl. mut.: … et actio quando inepte proponitur ut f. de [ (= cap. Forma proponendi exceptionem peremptoriam) (64ra–65vb) Lib. 3. Inc. mut. (cap. De notoriis criminibus): ] et probatur extra de regula veniens prope prin. Sequitur de manifesto videre … – Expl.: … in popularibus de hoc Extra De purg. can. c. tuarum. (= Ende Pars 3) (66ra–143vb) Lib. 4. Inc.: Formavit Deus hominem ad ymaginem et similitudinem suam … – Expl. mut.: … prefate ecclesie et quam tanquam suam [ (Reklamante: ancillam) (= cap. De natis ex libero ventre) Lit.: Schulte GQ II 148–152; K. W. Nörr, Die Literatur zum gemeinen Zivilprozeß, in: H. Coing (Hrsg.), Handbuch der Quellen und Literatur der neueren europäischen Privatrechtsgeschichte I. München 1973, 394. II (144[145]–163[164], I) B: Haar- und Fleischseite deutlich unterscheidbar, tw. fleckig, am Blattrand leicht gewellt. Lagen: 2.V163(164) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Die Blätter der ersten Lagenhälfte der ersten Lage mit roten Strichen am seitlichen Blattrand gezählt. S: Schriftraum 340 × 195. Zwei Spalten, von Tintenlinien gerahmt, zu 85 Zeilen auf Tintenlinierung. Zirkel stiche. Textualis formata (Littera Bononiensis). Korrekturen und Ergänzungen am Schriftrand tw. von gleicher, tw. von anderer Hand. 86 A: Cod. 19 Abwechselnd rote und blaue Paragraphzeichen, tw. mit Zackenverzierung (z. B. Bl. 152r). Tintenfarbene Randmarkierungen (z. B. Bl. 163rb), feine Unterstreichungen und flüchtige Zeigehände (z. B. Bl. 149ra). Repräsentanten, Überschriften nicht ausgeführt. Zu Beginn von Unterabschnitten dreizeilige Lombarden, abwechselnd in Rot und Dunkelblau mit Fleuronnée in der Gegenfarbe. Charakteristische saitenartige Vertikallinien, besetzt mit kleinen, tw. gestielten und ge punkteten Halbkreisen; Parallellinien als Konturierung, auch Fadenausläufer parallel geführt und gestaffelt in einfachen Voluten endend. Deckfarbenschmuck: zu Kapitelbeginn vierzeilige figürliche bzw. ornamentale Initialen (die schwarzen Konturlinien der goldenen Initialfelder spitz ausgezogen) mit wenigen Ausläuferblättern. Viele Parallelen zum Deckfarbendekor in T. I, aber nicht von derselben Hand stammend; die Büsten differenzierter mit welligem Haar, geraden Nasen und – speziell in der Profilansicht – stark betonten Augen (z. B. Bl. 155va), die Ornamentalinitialen (Blattformationen, z. B. Bl. 145rb, 148va) etwas aufwändiger ausgeführt. Der Deckfarbendekor in T. II ist sehr wahrscheinlich dem „Maestro della Crocifissione D“ zuzuschreiben, der beispielsweise auch in dem um 1340 entstandenen Decretum Gratiani München, BSB, Clm 23552 mit dem „Maestro del B 18“ sowie dem „Illustratore“ zusammenarbeitete; hierzu: G. del Monaco, I manoscritti miniati dell’Illustratore nella cultura figurativa del secondo quarto del XIV secolo, Diss. Bologna 2012, 62, 216–221 (Nr. 19), http://amsdottorato.unibo.it/5083/1/Tesi_del_Monaco.pdf; zu Clm 23352 s. auch U. Bauer-Eberhardt, Die illuminierten Handschriften italienischer Herkunft in der Bayerischen Staatsbibliothek 1: Vom 10. bis zur Mitte des 14. Jahrhunderts (Katalog der illuminierten Handschriften der Bayerischen Staatsbibliothek in München VI, 1: Text- und Tafelband). Wiesbaden 2011, 234–238 (Nr. 213). – Das Fleu ronnée in T. II ist oberitalienisch. S. R. 2 (144ra–163vb) ‹Iohannes Andreae›: Novella super librum sextum Decretalium, unvollständig (hier abgeglichen mit Druck: Venedig 1612). Inc. mut. (cap. De haereticis): ] simul citant cui obedire teneatur pro ordinario facit quia eius iurisdiccio firmior et antiquior cui specialis non derogat … – Expl. mut.: … quia tamen beneficia inter[pretamur (cap. De sententia excommunicationis) Lit.: Schulte GQ II 218–219; K. W. Nörr, Die Literatur zum gemeinen Zivilprozeß, in: H. Coing (Hrsg.), Handbuch der Quellen und Literatur der neueren europäischen Privatrechtsgeschichte I. München 1973, 378. (Ir–v) leer. P. A. / U. S. Cod. 19 PETRUS LOMBARDUS Perg. I, 108, I Bl. 385 × 270. Nordfrankreich (Raum Paris), 13. Jh. B: Dickes, an den Rändern abgegriffenes Perg., an den Rändern tw. beschnitten, vereinzelt bis zum Textrand mit geringem Textverlust, vereinzelt Nahtspuren. Bl. 38 keilförmig dreieckiges Pergamentstück heraus geschnitten. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + II4 + 4.IV36 + (IV–1)43 + (VI–1)54 + IV62 + III68 + 5.IV108 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Erste Lage unvollständig (Textverlust), urspr. wohl Quaternio. Es fehlen das Doppelbl. zw. Bl. 1 und 2 bzw. Bl. 3 und 4 sowie das Doppelbl. in der Lagenmitte zw. Bl. 2 und 3 (s. bei Inhalt). Gegenbl. zu Bl. 42 und 53 herausgeschnitten (ohne Textverlust). Sexternio Bl. 44–54 (vgl. bei S) nachträgliche Ein fügung nach Verlust von ein oder zwei Quaternionen. Nach Bl. 108 fehlt mindestens eine Lage (Textverlust). Lagenreklamanten. Am oberen Blattrand tw. Schriftreste (weggeschnitten, Zählung der Psalmen in rö mischen Zahlen). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 255 × 155. Zwei Spalten, seitlich von zweifachen Tintenlinien gerahmt, zu meist 67–70 Zeilen auf Tintenlinierung. Zirkelstiche am äußeren und inneren Blattrand. Textualis (Perlschrift), Bibeltext und Kommentar in unterschiedlicher Größe. Bl. 38ra–54vb Kommentar von anderer Hand in kleinerer Schrift 13. Jh. (Einfügung mit dünnerer Feder nach herausgeschnittenem Blatt zw. Bl. 37 und 38 bis Ende des Sex Cod. 19 87 ternio Bl. 54). Am seitlichen und unteren Blattrand zahlreiche Interlinear- und Randglossen von verschiede nen Händen 13. Jh. sowie neuzeitliche Randglossen (Bleistift, stark verblasst, z. B. Bl. 39r). A: Rote Unterstreichungen, am Beginn der Kommentarabschnitte rote und blaue Satzmajuskeln in Lombarden form, am Beginn der Psalmen und der Psalmenverse einzeilige, abwechselnd rote und blaue Lombarden mit Palmettenfleuronnée in der Gegenfarbe. Bl. 17vb (2), 37ra, 52rb, 55ra (2), 69va, 69vb vierzeilige (Bl. 55ra1 sechszeilige) Initialen in Deckfarbenmalerei der nordfranzösischen Schule: rosafarbener oder blauer Buch stabenkörper, gefüllt mit verschiedenfarbigen geometrischen Ornamenten oder Fabelwesen auf blauem oder blattvergoldetem Grund mit Ausläufern in Blattgold, Rosa und Blau. Blattgold, tw. abgeplatzt (z. B. Bl. 17vb) auf hellem Bolus. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. fleckig und zerkratzt. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck COME. IN PSALT. DAVID in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II1 C 6 (Bleistift) und Nr. 19 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 19. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 10030. – Steer 36. – Katalog ULB Tirol I 99, 281. (Ir–v) leer. (1ra–108vb) Petrus Lombardus: Commentarii in Psalmos, unvollständig: Ps 48–145 (PL 191, 465B–1271A). (1ra) Inc. mut.: ] cur timebo. Tercio dicitur que impiis et iustis reddenda sunt ibi … (= Schluss des Kommentars zum Tit. von Ps 48. PL 191, 465B). (1ra) Beginn von Ps 48. Text. Inc.: Audite hec omnes gentes auribus percipite omnes qui habitatis orbem … Kommentar. Inc.: Audite hec que dicturus sum. Hic attentos facit omnes Gregorius id est omnes iniqui, hec iam hodie … (PL 191, 465B). Zw. Bl. 1vb und 2ra Verlust eines Blattes, Schluss von Ps 48 und Beginn von Ps 49 fehlen. (2ra–vb) Ps 49,17–50,7. Zw. Bl. 2vb und 3ra Verlust eines Doppelbl. (Lagenmitte), Schluss von Ps 50, Ps 51 und Beginn von Ps 52 fehlen. (3ra–vb) Ps 52,6–54,8. Zw. Bl. 3vb und 4ra Verlust eines Blattes, Ps 54,9–21 fehlt. (4ra–108vb) Ps 54,21–145,4. Bei den herausgeschnittenen Gegenbl. zu Bl. 42 (zw. Bl. 37 und 38) und 53 (zw. Bl. 44 und 45) kein Textverlust. Sexternio Bl. 44–54 von anderer Hand (Text und Kommentar) als Ersatz für zwei Sexter nionen, ohne Textlücke. Im Ternio Bl. 63–68 keine Textlücke. (108vb) Expl. mut.: … quam in diversos seculi ambitus tendebant (= Kommentar). Beatus cuius Deus Iacob adiutor [ (Ps 145,4. PL 191, 1271A). (Ir–v) leer. W. N. 88 Cod. 20 Cod. 20 VETUS TESTAMENTUM Perg. I, 168, I Bl. 380 × 280. Entstehungsort unbestimmt, 12./13. Jh. B: Dickes Perg., tw. fleckig und löchrig, vereinzelt Nahtspuren, Bl. 121 und 125 seitlich stark beschnitten (380 × ca. 235 bzw. 230). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + III6 + IV14 + (IV+1)23 + (IV+1)32 + 17.IV168 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. 23 und 24 eingehängte Einzelbl. Bl. 48v (Ende der Lage 6) und 112v (Ende der Lage 14) Reklamanten, letztere stark beschnitten. Am oberen Blattrand zeitgenössische Foliierung in arabischen Ziffern 105–273 (Tinte), tw. beschnitten; Fehler in der Zählung: 111 ausgelassen. Am unteren Blattrand Foliierung 20. Jh. (Bleistift, hier berücksichtigt). Am oberen Blattrand Einteilung der Bücher von unter schiedlichen Händen. S: 1) Bl. 1r–24v: Schriftraum 295 × 195. Zwei Spalten, seitlich von zweifachen Bleistiftlinien gerahmt, zu 43 bzw. 44 Zeilen auf Bleistiftlinierung. Zirkelstiche. Karolingisch-gotische Mischschrift 12. Jh. Am Blattrand Kapitelzählung und Nota-Hinweise von späterer Hand. 2) Bl. 25r–96v: Schriftraum 290/300 × 175/190. Zwei Spalten, von zweifachen bis an den Blattrand reichen den Tintenlinien gerahmt, zu 45–47 Zeilen auf Tintenlinierung. Zirkelstiche. Textualis 13. Jh. 3) Bl. 97r–168v: Schriftraum 300 × 190. Zwei Spalten, von Tinten- bzw. Bleistiftlinien gerahmt, zu 43 Zeilen auf Tinten- bzw. Bleistiftlinierung. Textualis 13. Jh. von zwei Händen: 1) Bl. 97ra–120rb; 2) Bl. 120va– 168vb. Vereinzelt Randbemerkungen von späteren Händen. Bl. 14r, 79r und 101r Randglossen 14. Jh. A: Bl. 1ra–22vb schwarze (Bl. 2rb grüne, 3ra rote) einzeilige Lombarden, Bl. 7v–9r Textgliederung durch rote Buchstaben am Rand sowie Bl. 1v und 2r Angabe des Buches am oberen Blattrand in Rot und Grün. Bl. 3v flüchtige Federzeichnung eines Königs (Salomon) mit Zepter sowie rot ergänzter Krone und Zeigehand. Weitere Zeigehände Bl. 3v und 4r. Bl. 1va und 13ra elf- bzw. 17zeilige kopfstempelartig gespaltene Initialen in Rot, Blau und Grün mit routiniertem rot-blauem Palmettenfleuronnée, das in der Binnenfeldorganisation wenig Symmetrietendenzen zeigt; Bl. 12vb fünfzeilige rot gezeichnete und blau akzentuierte Rankeninitiale (mit Spangen) auf grünem Grund; Buchstabenkörper gespalten und blau gefüllt; die Rankenvoluten im Binnenfeld umschließen oben eine spitze, unten eine kelchförmig-gerundete Blüte; leicht bewegte, wenig schraffierte Rankenblätter. – Die Rankeninitiale und die beiden Fleuronnée-Initialen wohl von einer Hand 13. Jh. Bl. 25ra–119rb rote Auszeichnungsstriche und Überschriften, selten Durchstreichungen. Vereinzelt Zeige hände (Bl. 102v, 103v). Ein- bis dreizeilige (wein-)rote Lombarden und einige größere rote Initialen (bis zu 16zeilig, Bl. 25ra) mit Punktverdickungen und tw. kritzeligen geometrischen Aussparungen und Füllranken, jeweils mit Palmettenfleuronnée (auch als Besatz) in (hellerem) Blau, recht flüchtig und etwas disparat, aber wohl von einer Hand (13. Jh.); selten rotes Fleuronnée (z. B. Bl. 26r) von anderer Hand (auch 13. Jh.). In den Binnenfeldern der Lombarden u. a. Spiralen, bei den größeren Initialen einfache Ranken (Bl. 25ra), lanzett förmige Blätter (Bl. 38ra) und Palmetten; charakteristisch die fischgrätenartig angeordneten, stärkeren (Be satz-)Fäden mit kugeligen Köpfchen (Bl. 44ra, 66vb). Bl. 87va, 91ra, 93rb und 100rb sechs- bis achtzeilige rote Initialen mit Spangen am Buchstabenkörper und/oder Rosetten im Binnenfeld, die von Palmetten kränzen umgeben sein können; Bl. 93rb blauer Außengrund, Bl. 100rb größere, feiner ausgeführte Palmetten als Besatz. Bl. 50rb von einer anderen, ungeübteren Hand zehnzeilige Initiale E, u. a. mit ungelenkem, schwarz-rotem (Wappen-)Adler im oberen, gelblich gefärbten Binnenfeld; unklar, ob auch die zwölfzeilige hellrot(-blaue) Initiale Bl. 78rb, die in der Art einer Rankeninitiale angelegt ist, von dieser Hand stammt; Bl. 50rb und 78rb ebenfalls 13. Jh. Fleuronnée bis einschließlich Bl. 119rb, dann nur mehr rote Überschriften und einige rote Lombarden bis Bl. 146ra. Repräsentanten. Michaela Schuller-Juckes, Wien, sei für ihre Hinweise herzlich gedankt. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. stark abgerieben, insbes. an den unteren Kanten. Ecken mit braunem Leder verstärkt. Rücken: brau nes Leder, fünf einfache Bünde, mit Goldbordüren gerahmt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck S: S: Biblia Incipiens a III. Libro Regum in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Aufhe bung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signatur der ULB Tirol II`1 Cod. 20 89 C 16 (Bleistift) sowie Nr. 20 (Tinte) und Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 20. Bl. Iv Abklatsch des Stempels der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Steer 36. – Katalog ULB Tirol I 99, 281 – Baroffio 324, Nr. 16708. (Ir–v) leer. (1ra–168vb) Vetus Testamentum, unvollständig. Am Ende der einzelnen Bücher meist Angabe einer Versanzahl. (1ra) 2. Regum. Inc. mut.: ] de manu Egyptii et interfecit eum hasta sua. Hec fecit Banaias filius … (2 Rg 23,21). – (1va) 3. Regum. – (13ra) 4. Regum. – (24vb) leer. (25ra) Iudices. – (36rb) Ruth. (38ra) Hieronymus: Prologus in librum Iudith (BS VIII 213–214. Préfaces 35, Nr. 2. – Stegmüller RB Nr. 335). – (38ra) Iudith. (43vb) Hieronymus: Prologus in librum Esther (BS IX 3–4. Préfaces 36, Nr. 3 und 4. – Stegmüller RB Nr. 341 und 343). – (44ra) Esther. (49va) 1. Maccabaeorum. – (66rb) 2. Maccabaeorum. (77rb) Hieronymus: Prologus in libros Salomonis (BS IX 3–5. Préfaces 118, Nr. 1. – Stegmüller RB Nr. 457). (77va) ‹Hieronymus›: Prologus in libros Salomonis (BS IX 7–9. Préfaces 119–120, Nr. 3: Commenta rius in Eccl. – Stegmüller RB Nr. 456). (77vb) Hieronymus: Prologus in libros Salomonis (BS XI 6. Préfaces 118–119, Nr. 2. – Stegmüller RB Nr. 455). (78ra) Proverbia. – (87va) Ecclesiastes. – (91ra) Canticum canticorum. – (92vb) Sapientia. (99va–b) Hieronymus: Prologus in librum Ecclesiastici (BS XII 145–147. – Stegmüller RB Nr. 26). – (99vb) Sirach (Ecclesiasticus) mit cap. 52. (119va) Hieronymus: Prologus in librum Esdrae (BS VIII 3–7. Préfaces 33–34, Nr. 1. – Stegmüller RB Nr. 330). – (120ra) 1. Esdras. – (123vb) 2. Esdras (Nehemias). In der Hs. als Nehemias, unterteilt in zwei Bücher: (123vb) 1. Nehemias = 2. Esdras 1–7,69. (126va) 2. Nehemias = 2. Esdras 7,70–13,31. (129va) Hieronymus: Prologus in librum Iob (BS IX 69–74. Préfaces 38, Nr. 1. – Stegmüller RB Nr. 344). – (130ra) Iob. (140va) Hieronymus: Prologus in librum Tobiae (BS VIII 155–156. Préfaces 35, Nr. 1. – Stegmüller RB Nr. 332). – (140vb) Tobias. (145ra) Hieronymus: Prologus in libros Paralipomenon (BS II 546–547. Préfaces 30–31, Nr. 1. – Steg müller RB Nr. 328). (145va) Hieronymus: Prologus in libros Paralipomenon (Préfaces 31–32, Nr. 2. – Stegmüller RB Nr. 327). (146ra) 1. Paralipomenon. – (158rb) 2. Paralipomenon. (168vb) Expl. mut.: … Steteruntque viri quos [ (2 Par 28,15) (Ir–v) leer. A. P. / U. S. 90 Cod. 31 Cod. 31 ALDOBRANDINUS DE TOSCANELLA. EXPOSITIO SUPER AVE MARIA. GODEFRIDUS HERILACENSIS Perg. I, 130, I Bl. 320 × 240. Süddeutscher Raum (?), 14. Jh. B: Perg. tw. eingerissen, löchrig, vereinzelt Nahtspuren, Haar- und Fleischseite deutlich unterscheidbar. Lagen: (I–1)I (Vorsatzbl. Pap. 18. Jh.) + 10.IV80 + (IV+2)91(90) + 5.IV131(130) + (I–1)I* (Nachsatzbl. Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 85 ausgelassen. Bl. 90 und 91 eingehängte Einzelbl., urspr. Teil einer eigenen Lage, zw. Bl. 90 und 91 fehlt mindestens ein Blatt (Textverlust). Nach Bl. 131 fehlt mindestens eine Lage (Textverlust). Lagenreklamanten, tw. gerahmt oder unterstrichen. S: Schriftraum 250 × 185. Zwei Spalten, von Tintenlinien gerahmt, zu einheitlich 33 Zeilen auf Tintenlinierung. Zirkelstiche. Textualis. Korrekturen und Nachträge von späterer Hand. A: Rote Auszeichnungsstriche und Paragraphzeichen. Abwechselnd rote und blaue ein- bis vier-, meist zwei zeilige Lombarden mit einfachem Fleuronnée in der Gegenfarbe. Bl. 1ra und 49ra jeweils fünfzeilige blaue Initiale mit kopfstempelförmigen Aussparungen und etwas ungelenkem rotem Knospenfleuronnée im Bin nenfeld, Bl. 1ra als Besatz Perlenpyramiden, Bl. 92ra fünfzeilige rot-blaue Initiale mit kopfstempelförmigen Aussparungen und einfachem rotem Fleuronnée im Binnenfeld; Ähnlichkeit mit der zweiten Floratorenhand in Cod. 38 (dat. allerdings ins 15. Jh.). S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. leicht abgerieben. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S: und Signaturschild (s. bei G), im zweiten Gold druck EXPOSIT. IN CREDO AVE MAR. ET PATER NOSTER in Goldrahmung. Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Bl. 1r (stark beschnitten) sowie Bl. 48vb Besitzvermerk 15. Jh.: Iste liber est monasterii sancte Marie in Novacella T 13. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorüberge henden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 D 9 (Tinte, Bleistift) sowie Nr. 31 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 31. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den ita lienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: E. Bauer (Hrsg.), Paternoster-Auslegung, zugeschrieben Jakob von Jüterbog, verdeutscht von Heinrich Hal ler (Lunder germanistische Forschungen 39). Lund, Kopenhagen 1966, 17, 66–67, 83 u. ö. – B.-G. Guyot, A propos de quelques commentaires sur le Pater noster. Revue des Sources philosophiques et théologiques 53 (1969) 245–255, hier 248–250. – Kaeppeli Nr. 133. – Stegmüller RB Nr. 1105 und 3886. – Bloomfield Nr. 8085. – Katalog ULB Tirol I 118, 282. – E. Bauer (Hrsg.), Godefridus Herilacensis „Expositio super orationem dominicam“ in der Übersetzung Heinrich Hallers (Analecta Cartusiana 263). Salzburg 2008, 7–8. – Kustatscher–Korenjak 158. 1 2 (Ir–v) leer. (1ra–48rb) ‹Aldobrandinus de Toscanella›: Scala fidei (Schneyer 1, 250, Nr. 350. Kaeppeli Nr. 133. Stegmüller RB Nr. 1105. Distelbrink Nr. 197). (48va–b) leer bis auf Besitzvermerk (s. bei G). (49ra–91vb) Expositio super Ave Maria, unvollständig. (49ra) Inc.: Ave [Maria] gratia plena dominus tecum (Lc 1,28) benedicta tu in mulieribus (Lc 1,42). Dixit angelus ad virginem de celo missus. Et benedictus fructus ventris tui. Adicit Elyzabeth spiritu prophetico inspirata … – Expl.: … tercio de salutacionis compendio. Primo igitur dicendum est de angeli officio o (?). (49vb) Inc.: Missus est Gabriel angelus a Deo. Luc. 1 (26). In hoc verbo Gabriel angelus sacra menti regalis conscius ac Dei fore virginis paranymphus describitur … – Expl.: … decuit ipsum Cod. 31 91 secreciori consilio pertractari. Et sic patent octo ministeria que Gabriel exhibuit circa virginis uterum fecundandum. (54ra) Inc.: Secundo idem angelus octo officia exercuit cir[ca] filium virginis declarandum. Primum erat officium docendi unum Ioseph dubitantem … – Expl.: … volavit unus ad me de Seraphyn etc. Postquam dictum est de angeli officio dicendum est de virginis vocabulo. (55va) Inc.: Nomen virginis Maria. Luc. 1 (27). Nulla res potest amari nec laudari nisi prius cognoscetur dicente Augustinus (!) de trinitate … – Expl.: … visione preclara fruicione beata intencione eterna. (58rb) Inc.: Nomen virginis Maria. Luc. 1 (27). De dulcissimo nomine ipsius Marie adhuc am plius videamus. Est notandum quod nomen huius dulcissime virginis est a Deo revelatum … – Expl.: … huius nomen gloriosum wlgatum est in omni terra et in omni regione celestium et infernorum. (60va) Inc.: Nomen virginis Maria. Luc. 1 (27). De nomine huius ineffabilis virginis adhuc est plus dicendum. Et est notandum quod hec virgo postquam Christum concepit reperitur in scrip tura … – Expl.: … que cum sit omnibus superior servicio se subiecit omni. (65va) Inc.: Nomen virginis Maria (Lc 1,27). Prius dictum est de nomine. Nunc dicendum est de hoc vocabulo Maria. Et est notandum quod IIIIor genera volatilium solent nidos suos ornare … – Expl.: … Postquam dictum est de angeli officio et de virginis vocabulo. Nunc dicendum est de salutacionis conpendio. (67rb) Inc.: Ave gratia plena dominus tecum. Luc. 1 (28). Si quis princeps vel rex vel magnus dominus vult destinare nuncium vel legatum ad summum principem vel regem vel dominum magnum pro magna et ardua causa … – Expl.: … Non diligamus verbo nec lingua scilicet tantum sed opere et veritate. (69ra) Inc.: Gracia plena (Lc 1,28). Dictum est prius quomodo dulcissima virgo Maria est gloriose salutata in hoc verbo Ave. Nunc dicendum est quomodo graciose est adornata … – Expl.: … que in Maria clarius et plus Christianus quam in aliquo homine refulserunt. (71va) Inc.: Gracia plena (Lc 1,28). Notandum quod sicut sunt VII utilitates sive efficacie quas gracia confert anime de quibus dictum est in sermone precedenti dicendum est … – Expl.: … habebit amicum regem. Quam amiciciam nobis concedat. (73rb) Inc.: Gracia plena (Lc 1,28). Prius dictum est quod gracia habet VII efficacias quas in anima facit et quod sunt septem preparaciones quibus homo ad graciam se disponit … – Expl.: … sive possumus sive habemus ad quam gloriam nos perducat. (75vb) Inc.: Gracia plena (Lc 1,28). Prius dictum est quomodo Maria fuit graciose salutata quod notabatur in hoc verbo Ave … – Expl.: … denique necessitatis amplissimo miseretur affectu. (79vb) Inc.: Dominus tecum. Luc. 1 (28). Secundum ordinem naturalem in mundo corpora magis conformia in natura sint magis propinqua in situ … – Expl.: … Iste ergo est dominus qui fuit tecum o Maria. (82ra) Inc.: Dominus tecum (Lc 1,28). O Maria dominus tecum te associans tecum te custodiens tecum te glorificans … – Expl.: … mecum fac ut amor tuus semper sit mecum. (84va) Inc.: Benedicta tu in mulieribus. Luc. 1 (42). Illud digne commendatur quod in se con sideratum bonum apparet et aliis comparatum excellit … – Expl.: … pronunciando et dicendo Magnificat anima mea domino. (89rb) Inc.: Benedicta tu in mulieribus (Lc 1,42). Si volumus amplius scire quod multipliciter est sanctissima virgo domina Maria benedicta inspiciamus … – Expl. mut.: … Et quod bona domine Marie sunt precellentiora omnia et in Christo singulariter bene[ (= Rom, Vaticana, Cod. Vat. lat. 1287, S. 42a, Z. 7) (91ra) (Lc 1,42). Inc. mut.: ] est in multis pro nobis constituta … (= Rom, Vaticana, Cod. Vat. lat. 1287, S. 42a, Z. 9 von unten) – Expl. mut.: … Domina Maria eciam corpus infantis suaviter at tractat quia quicquid facit circa hominem totum est suavitas et pietas [ (= Rom, Vaticana, Cod. Vat. lat. 1287, S. 43a, Z. 3 von unten) 92 Cod. 38 Parallelüberlieferung: Klosterneuburg, Stiftsbibl., CCl 774, Bl. 150r–172v (unvollständig, Text nur bis Neustift, Stiftsbibl., Bl. 77v, Z. 22, erhalten); ab Bl. 49vb: Rom, Vaticana, Cod. Vat. lat. 1287, S. 1. Zu einer möglichen Zuweisung an Siboto von Wien vgl. Ch. Jörg, Untersuchungen zur Bücher sammlung Friedrichs von Amberg. Ein Beitrag zur franziskanischen Geistesgeschichte des Spät mittelalters. Zeitschrift für Schweizerische Kirchengeschichte / Revue d’histoire ecclésiastique suisse 69 (1975) 1–264, hier 95–96. 3 (92ra–131vb) ‹Godefridus Herilacensis› (= Ps.-Iacobus de Paradiso): Expositio super orationem dominicam, unvollständig (Ed. Bauer, Paternoster-Auslegung 84–226. – L. Meier, Die Werke des Erfurter Kartäusers Jakob von Jüterborg in ihrer handschriftlichen Überlieferung [Beiträge zur Geschichte der Philosophie und Theologie des Mittelalters 37, 5]. Münster/W. 1955, Nr. 24. Steg müller RB Nr. 2608, 3886, vgl. auch Nr. 1551, 11642. Bloomfield Nr. 8085 bzw. 8203). Expl. mut.: … proximo tuo nocenti te et tunc deprecanti tibi peccata tua solventur. Hom[o homini servat (Sir 28,2–3). Parallelüberlieferung: u. a. Innsbruck, ULB Tirol, Cod. 24, Bl. 120ra, Cod. 365, Bl. 1ra, Cod. 705, Bl. 11va; Klosterneuburg, Stiftsbibl., CCl 293, Bl. 216ra. Zur Verfasserfrage s. Bauer, Paternoster-Auslegung 66–67, Guyot, Commentaires und Bauer, Godefridus Herilacensis 7–9. Wie Innsbruck, ULB Tirol, Cod. 24 zählt Neustift, Stiftsbibl., Cod. 31 zu den Hs., die die längere Textversion überliefern (CV-traité-Typus, s. Guyot, Commentaires 248–249). (Ir–v) leer. A. P. Cod. 38 THOMAS DE AQUINO Perg. I, 272, I Bl. 345 × 260. Süddeutscher Raum (?), 15. Jh. B: Dickes Perg., vereinzelt Löcher und Nahtspuren, tw. unregelmäßig. Bl. 127v stark verschmutzt. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 34.IV269(272) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nach satzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am Beginn und am Ende fehlt jeweils wohl mindestens eine Lage (Textverlust). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 115, 161 und 171 doppelt angeführt. S: Schriftraum 240/250 × 180/185. Zwei Spalten, von vielfach bis an den Blattrand reichenden Tintenlinien gerahmt, zu 38–40 Zeilen. Zirkelstiche. Bastarda, Textanfänge in Textualis formata (Textura). A: Rote Auszeichnungsstriche, Unterstreichungen, Rahmungen, Über- und Schlussschriften sowie Bl. 1r–2r, 7r–12r, 139ra, 141ra, 144v, 171rb, 261ra, 268ra am oberen Blattrand Angaben der jeweiligen Quaestio. Vereinzelt auch blaue Überschriften (Bl. 70vb). Rote und blaue Paragraphzeichen. Vereinzelt Zeigehände mit Ärmeldraperie (z. B. Bl. 142ra). Rote und blaue ein- bzw. zweizeilige Lombarden, tw. mit Punktverdic kungen, Fadenausläufern (tw. in Achterschlinge gelegt, tw. gestrichelt), (kopfstempelförmigen) Aussparun gen und Knospenfleuronnée in der Gegenfarbe; Bl. 81r und 89r gelblich-braune (verblasste?) Lombarden und Paragraphzeichen. Oberlängen in der ersten bzw. Unterlängen in der letzten Zeile bisweilen cadellenar tige Dehnungen, tw. mit Blattwerk und Knospenfleuronnée gefüllt, vereinzelt in Schwarz und Rot. Bl. 21rb Ausläufer mit bärtigem Profilkopf, Bl. 25ra u. a. mit plastisch herausgearbeitetem Kopf. Am oberen Blatt rand Dehnungen vereinzelt beschnitten. Bl. 144va vierzeilige blaue Initiale P mit rotem Initialfeld, das mit silbernem Knospenfleuronnée bemalt ist; Bl. 161Irb zweizeilige blaue Lombarde mit dunkelrotem und braunem Fleuronnée, Perlenpyramiden als Besatz. Repräsentanten. Fleuronnée von zwei Händen, die sich Pars 1 und 2 aufgeteilt haben; die zweite Hand vgl. Cod. 31 (Ent stehungszeit der Hs. 14. Jh.). S. R. 93 Cod. 52 E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. abgerieben und beschädigt. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, oben schadhaft, fünf einfache Bünde, mit Goldbordüren gerahmt. Im ersten Feld Golddruck M: S:, im zweiten SUMA ANGELICI PARS II DE INCARNATIONE in Goldrahmung, im dritten Feld Signaturschild (s. bei G). Schnitt rot gefärbt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`2 G 13 (gestrichen, korr. zu 12) (radiert, Tinte), II 1 E 6 (Bleistift) sowie Nr. 38 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 38. Bl. 1r Stempel der ULB Tirol 19 Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat ab gegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Shooner II 51. – Katalog ULB Tirol I 130, 282. (Ir–v) leer. (1ra–269vb) Thomas de Aquino: Summa theologica, Pars 3. Quaestiones de incarnatione dni, unvoll ständig: Beginn und Schluss fehlen (Ed. Thomas de Aquino, Opera omnia iussu impensaque Leonis XIII. P. M. edita. Bd. 11. Romae 1903, hier ab 134). (1ra) Pars 1 (qu. 1–59), unvollständig: hier ab qu. 8, art. 6. Inc. mut.: ] non procedit ab ore vestro sed si quis bonus est ad edificacionem fidei ut det graciam audientibus. Ergo videtur quod eciam alii quam Christo competat esse ecclesie capud. Propterea Christus ex eo quod preest ecclesie non solum dicit capud … – Expl.: … quia unus et idem est Deus et homo dominus Ihesus Christus de cuius incarnacionis misterio ad presens dicta sufficiant (gestrichen: Explicit tercia pars sancti Thome). Schlussschrift: Explicit prima pars huius tercie partis ffratris Thome de Aquino scilicet de salvatore nostro et misterio incarnacionis. (144rb) Pars 2 (qu. 60–90), unvollständig: hier bis qu. 85, art. 4. Tit.: Incipit secunda pars eiusdem tercie partis Summe Thome que est de sacramentis. – Incipit (gestri chen: quarta) secunda pars (gestrichen: Summe s. Thome). – Inc.: Post consideracionem eorum que pertinent ad misteria verbi incarnati considerandum est de ecclesie sacramentis … – Expl. mut.: … in libro de memoria sed penitencia est de preterito ut dictum est ergo penitencia est in memoria sicut in subiecto. Preterea nihil agit [ (Ir –v) leer. A. P. / U. S. Cod. 52 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 273 Bl. 290 × 205. Süddeutscher Raum (Tirol?), T. I: Ende 14. Jh., T. II: 1396. B: Zwei Teile: I (1–39), II (40–273). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. E: Schmuckloser gotischer Einband: rot gefärbtes Leder über dickem Holz. Süddeutscher Raum (Tirol?), Ende 14./Anfang 15. Jh. Leder stark abgerieben und zerkratzt, Holz an den Ecken tw. freiliegend. VD und HD gleich: Spuren von je fünf kleinen runden Beschlägen. Am VD oben mittig aufgeklebtes Papiertitelschild Ysidorus de summo bono. Item questiones sentenciarum Boneventure secundi et quarti librorum. Item similitudines rerum XI, daneben und darunter Reste von Titelaufschriften (Tinte auf Leder), u. a. Ysidorus de summo bono, auf stark abgeriebenem Leder XI. Urspr. zwei Langriemen-Stoff-Metall-Schließen, heute nur mehr obere erhalten. Kanten abgeschrägt und gerade. Rücken: vier Doppelbünde, im ersten Feld aufgeklebtes Papierschild M: S: 1396, im zweiten Papierschild Ysidori Theologia, im dritten Signaturschild (s. bei G), im fünften Papier 94 Cod. 52 schild aufgeklebt (ohne Beschriftung). Kapitale mit Lederband umstochen. Am Spiegel des VD Verse und Notizen (s. bei Inhalt), Exlibris und Signaturen (s. bei G), am Spiegel des HD Besitzvermerk (s. bei G) und Federproben. In den Ecken sowie in der Mitte zur Verstärkung jeweils beschriebene Pergamentstücke auf geklebt, tw. mit Notation. G: Bl. 2r Besitzvermerk 15. Jh. Iste liber est monasterii s. Marie v[irginis] ad Novacellam. Am Spiegel des HD Besitzvermerk 15. Jh. Iste liber est s. Marie v[irginis] in Novacella. Bl. 2r Besitzvermerk 18. Jh. Ad Collegium Neocell[ensem]. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 F. 9 (Tinte, Bleistift) und Nr. 52 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 52. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe über lassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 161, 282. – Stefani 94. I (1–39) B: Pap. bes. am unteren Blattrand tw. verschmutzt (u. a. Tintenflecken), Bl. 1 beinahe lose. Bilder und Nach weise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 589, T. I (dat. 1395). Lagen: (II–1)3 (Vorsatzbl.) + 3.VI39. Gegenbl. zu Bl. 3 als Spiegel auf dem VD aufgeklebt. In den Lagenmitten beidseitig beschriebene Pergamentfälze. Am Ende der zweiten und dritten Lage Kustoden in arabischen Ziffern. S: Schriftraum 195/210 × 130. Zwei Spalten, von Tintenlinien gerahmt, zu 35–40 Zeilen. Bastarda von zwei Händen: 1) Bl. 2va–3rb und 38vb–39vb; 2) Bl. 4ra–38va. A: Rote Auszeichnungsstriche, Unter- und Durchstreichungen, Über- und Schlussschriften sowie Angabe des jeweiligen Buches am oberen Blattrand. Bl. 4ra rote Zeigehand. Ein- bis dreizeilige rote Lombarden, bis weilen mit Punktverdickungen und einfachen Fadenausläufern. Rote Tinte bisweilen verschmiert. Bl. 4ra in Rot und Tintenfarbe kopfstempelförmig gespaltene P-Initiale (Höhe ca. 80 mm). Repräsentanten. 1 (VDS–1ra) Notizen und Verse. 1) Notiz (zentriert, rot). Lib[…]. – Nota. – In annuncciacione (!) dominica Deus homo factus est et die eodem passus, Ioseph venditus, Isaac ymolatus, Adam formatus, casus demonum, Iohannes Baptista decollatus, Petrus a vinculis solutus Abel interfectus Iacobus passus infernus spoliatus latro in paradisum locatus offe[…] Melch[isedech?]. Abrahe. 2) Versus de sanctis Iohanne Baptista et Iohanne ev. (in zwei Spalten). Nota. – Inc.: De duobus filiis summe deitatis (recte: veritatis) / Quos dedit ecclesie pater pietatis / Quos eorum habeat plus auctoritatis … – Expl.: … […] valeat videatur clare. Parallelüberlieferung: Heidelberg, UB, Pal. lat. 965, Bl. 241va. 3) Federprobe am unteren Blattrand. 4) Notiz zu den Unterschieden zw. der griechischen und der lateinischen Kirche. Nota. – Inc.: In hoc differunt Greci a Latinis quia Greci similiter omnes dicunt … – Expl.: … dicunt tum calice stagni argenti vel auri debere agi quia lignum est multum porosum. Darunter gestrichen: traditor autem dedit eis signa dicens quam […] scilicet. (1rb–v) leer. (2r) Besitzvermerk und Stempel (s. bei G). Notiz zum Inhalt (18. Jh.) Theologia. Isidori (gestri chen: libri) Anno 1396. 2 (2va–38va) Exzerpte aus Isidorus Hispalensis: De summo bono seu Sententiarum Lib. 1–3 (PL 83, 653–738 bzw. CCSL 111, 7–330. – CPL Nr. 1199. Bloomfield Nr. 4873, 4888 und 5854 sowie Bloomfield, Suppl. Nr. 5854). Cod. 52 95 (2va–3rb) Register. Tit.: Incipit (gestrichen: dominica se[…]; darüber, ebenso gestrichen: dicta mina [?]) registrum super librum Ysydori De summo bono. (3v) leer. (4ra) Prolog. Inc.: Premissis in nomine Christi librorum tabulis sive capitulorum tytulis secuntur ex eis … – Expl.: … forme verborum autorum (!) ydemptitate. (4ra) Text. Inc.: Solus Deus dicitur immortalis quia solus est incommutabilis nam et omnia (recte: anima) moritur dum de ore autem (recte: deserente) Deo de bono in malum mutatur … – Expl.: … de quibus bene per prophetam dicitur Ducunt in bono dies suos et in puncto ad inferna descendunt etc. Schreibervers: Finis adest operis mercedem posco laboris etc. (Colophons Nr. 21270). Schlussschrift: Explicit Ysidorus de summo bo[no]. (4ra–13ra) Lib. 1. – (13ra–24rb) Lib. 2. – (24rb–38va) Lib. 3. Parallelüberlieferung dieser Exzerpte: u. a. Innsbruck, ULB Tirol, Cod. 120, Bl. 1r; Cod. 262, Bl. 1v; Cod. 410, Bl. 38v; Cod. 449, Bl. 202ra und Cod. 608, Bl. 1ra. 3 3.1 (38vb–39ra) Exzerpte (u. a. aus Flavius Iosephus: De antiquitatibus und De bello Iudaico, Hieronymus: Epistola ad Helvidium, Thomas de Aquino: Catena aurea in Lucam), in drei Kapitel gegliedert. (38vb) Inc.: Ab Adam usque ad diluvium duo milia VI centi etc. Iosephus primo libro Antiquita tum migraverunt X tribus de Iudea … – Expl.: … meam terram rediit liberatus a servicio Babil orum Iosephus I libro de Iudaico bello. 3.2 (38vb–39ra) Inc.: Nulla obstetrix nulla muliercularum sedulitas intercessit ipsa pannis involvit infantem … – Expl.: … pre filiis hominum et matris obediens et moribus facetus. 3.3 (39ra) Tit.: De baptismo. – Inc.: In fontibus ecclesiarum non in privatis domibus debet baptisma concedi … – Expl. mut.: … puniri. In Clementinis de supremo (?) presenti [ 4 (39rb–vb) ‹Iacobus de Voragine›: Legenda aurea sanctorum, sive Lombardica historia, cap. 35: De ieiunio quattuor temporum (Ed. Graesse). Anschließend Schluss von cap. 34: De Quadragesima: Quare autem non observamus nostrum ieiunium … ut digni valeamus comedere agnum vite. Schlussschrift: Amen dicat omnis homo. II (40–273) B: Bl. 46v–47r Tintenflecken. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Lagen: II43 + 5.VI103 + (VI+III)121 + 6.VI193 + 2.V213 + 5.VI273. In Lage 7 Ternio eingebunden (Bl. 106–111). In den Lagenmitten beidseitig beschriebene Pergamentfälze. Am Lagenende Kustoden in arabischen Ziffern, bei letzter Lage auf dem ersten Blatt. Bl. 44–105 am oberen Blattrand mittig zeitgenössische Foliierung in roten arabischen Ziffern. S: Schriftraum 210 × 130/140. Zwei Spalten, von Tintenlinien gerahmt, zu 30–36 Zeilen. Bastarda, dat. 1396 (Bl. 270vb). Bl. 43v (Register) dreispaltig mit jeweils eigener zusätzlicher Spalte für die Blattzahlen, jeweils von zweifachen roten Tintenlinien begrenzt. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Überschriften. Bl. 204rb–264rb am Blatt rand Hervorhebung einzelner Lemmata in Rot. Bl. 264vb Zeigehand. Ein- bis vierzeilige (Bl. 202ra fünf zeilige) rote Lombarden, tw. mit Punktverdickungen, Bl. 44ra dreizeilig mit einfachem Dekor im Binnenfeld (rot) und als Besatz (tintenfarben); Bl. 112ra fünfzeilige rot-tintenfarbene Lombarde (als kopfstempelförmig gespaltene Lombarde angelegt, tintenbrauner Buchstabenteil C einfach darübergemalt). Bl. 110v–111r (Register) jeweils seitenhohes Kürzel DI (für „Distinctio“) und Abschnittsbuchstaben in Rot. 96 5 Cod. 52 (40r–43r) leer. (43v–105vb) Bonaventura (‹Bonaventura abbreviatus›): Super II Sententiarum (Stegmüller RS Nr. 142, II. Distelbrink Nr. 187. G. E. Mohan, Initia operum Franciscalium. Franciscan Studies 37 [1977] 313–314). (43v) Register mit Blattzahlangabe. Tit.: Incipit registrum capitulorum secundi libri Sentencia rum sequentis secundum numerum. (44ra) Text. Tit. (am oberen Blattrand): Liber Sentenciarum secundus. Tit. (am Textbeginn): Questiones Boneventure de lapsu. – Inc.: Queritur utrum angelus in primo instanti creacionis fuerit malus actu proprie voluntatis … – Expl.: … nichil intelligere possunt de salute beatorum etc. Expliciunt. (106r–110r) leer. 6 (110v–201ra) ‹Humbertus de Prulliaco›: Extractiones super IV librum Sententiarum Petri ombardi (Stegmüller RS Nr. 380, IV. Vgl. Glorieux Nr. 365c). L (110v–111r) Register. Tit.: Incipitur particio distinccionum super 4m librum Sentenciarum. (111v) leer. (112r) Text. Inc.: Circa quartum librum Sentenciarum primo queritur quid sit sacramentum. Dicendum quod sacramentum quattuor modis solet describi vel definiri. Primo sic sacramentum est sacre rei signum … – Expl.: … ad gaudia eos secum trahit eterna. Ad que nos perducat Ihesus Christus Amen. Schlussschrift: Expliciunt extracciones super quartum librum Sentenciarum continentes veri tatem etc. (201rb–vb) nur Rahmung. 7 (202ra–270vb) ‹Iacobus de Lausanna›: Compendium moralitatum (Kaeppeli Nr. 2090. Bloom field Nr. 82. Glorieux Nr. 345aa. Stegmüller RB Nr. 3887. Thorndike–Kibre 9. Walther, Initia Nr. 70. Welter 349 Anm. 38). Inc.: Abicit mundus pauperes et honorat divites. Nota Augustinus dicit super quod corvus est il lius nature quod pullos suos … – Expl.: … propter carnem quia pennas virtutum non habent etc. Schlussschrift mit Datierung: Explicit liber iste. Sit laus et gloria Christo Amen. Anno domini 1396o in vigilia assumpcionis s. Marie etc. (14. August 1396). (271r–273v) leer. 8 (HDS) Besitzvermerk (s. bei G). Fragmente (s. bei E). Federproben: homo natus – de peccator ibus quare de C. circa principium – dileccio creature impedit dileccioni di[cit] Augustinus ponit Extra De aceto quare de d. 9 (Fälze) Fragmente aus ‹Alexander de Villa Dei›: Doctrinale (Ed. Reichling [Monumenta Ger maniae paedagogica XII]. Berlin 1893, 7–178). Mit Interlinearglossen. Perg., Textualis formata (Textura) Anfang 14. Jh. auf Tintenlinierung. Interlinearglossen in Textualis cursiva von zeitgenössischer Hand. Rote Auszeichnungsstriche, Paragraphzeichen und Initialen. Enthält u. a.: Victrices turbe victricia tela tulere (V. 693). – Si brevis est e vel i producitur atque facit di (V. 723). – Dat lepus hic tenet intercus genus omne vetusque (V. 663). – Hoc monasilla (!) dant ut thus [sed dicitur] hec grus (V. 665). – preteriens in vi dat itum nullumve supinum (V. 741). – dant duo bos impos corapos custosque sacerdos (V. 651). – in triplici genere decet adiectiva notare (V. 683). – ag- vel cognosco dat itum sepelire sepultum (V. 763). – mons pons fons hic dant Mars dens cal[s quoque] (V. 675). – continue iuncta si non lex non tenet [ista] (V. 727). U. S. Cod. 53 97 Cod. 53 THOMAS EBENDORFER Pap. I, 348 Bl. 285 × 210. Wiener Raum (?), Mitte 15. Jh. (u. a. 1452). B: Pap. am Beginn und am Schluss des Buchblocks fleckig, bes. bei erster Lage unterer Blattrand eingerissen, Bl. 74 beinahe lose. Blattränder bisweilen eingerissen, Riss Bl. 218 mit Papierstreifen geflickt. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Identische Wasserzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 407, T. I ([Wien], dat. 1455); Wien, ÖNB, Cod. 3719 (dat. 1453). Lagen: 1I + 29.VI348. Vorsatzbl. eingehängtes Einzelbl., beschriebener Pergamentfalz auf Bl. Iv und Bl. 13r aufgeklebt. In den Lagenmitten Pergamentfälze, tw. beschrieben (tw. in dt., tw. in lat. Sprache). Lagenreklamanten, tw. stark beschnitten, vereinzelt rot gerahmt. Bl. 110–290 am Lagenbeginn bisweilen Reste von Kustoden in römischen Zahlen in Rot, stark beschnitten. Am oberen Blattrand Foliierung durch die ULB Tirol 19. Jh.; Fehler in der Zählung: 12 ausgelassen, 325 doppelt angeführt. S: Schriftraum 190/205 × 140/175. Zwei Spalten, von feinen Tinten-, Bl. 254r–275r von Blindlinien gerahmt, zu 40–49 Zeilen. Bastarda von drei Händen: 1) Bl. 1ra–253vb, dat. 1452 (Bl. 237ra); 2) Bl. 254ra–301vb; 3) Bl. 302ra–348rb. A: Rote Auszeichnungsstriche, Durch- und Unterstreichungen, Rahmungen und Kapitelzählungen am Blatt rand. Zwei- bis siebenzeilige rote Lombarden, tw. mit Punktverdickungen, einfachen Ausläufern und etwas Fleuronnée (Bl. 205ra mit schwarzem Knospenfleuronnée). Bl. 57va und 58rb drei- bzw. vierzeilige blaue Lombarde mit einfachem rotem Fleuronnée. Bl. 1ra zwölfzeilige rote Initiale mit kopfstempelförmiger Aus sparung. Am Blattrand vereinzelt rote Federzeichnungen (u. a. Bl. 152ra Vogelkopf im Profil [von geübter Hand], Bl. 285ra Christuskopf mit Nimbus). Ab Bl. 338rb Initialen nicht ausgeführt, Bl. 66v–73v keine Rubrizierung. Zahlreiche rote und schwarze Zeigehände. Am oberen Blattrand auf dem Verso jeweils Fest bzw. Sonntag angegeben, auf dem Recto die jeweilige Sermozahl (ausgenommen Bl. 106v und 107r), tw. von Titel im Fließtext abweichend. Repräsentanten. E: Gotischer Einband: rot gefärbtes Schafleder über Holz mit Blindlinien. Neustift, Mitte 15. Jh. Leder an den Kanten und insbes. am HD stark beschädigt. Rücken im 18. Jh. mit braunem Leder überklebt. Am VD und HD je zwei aus einfachen Streicheisenlinien gebildete Rahmen. Mittelfeld durch einfache Streicheisenlinien diagonal unterteilt. Spuren von je drei, urspr. wohl fünf runden Beschlägen und von zwei Langriemen-Leder-Metall-Schließen. Am VD oben aufgeklebtes Pergamenttitelschild mit Aufschrift (in Textura 15. Jh.) Haselbach de tempore gemali (!), darunter Reste eines aufgeklebten Pergamentschildes. Am HD oben Bohrloch, urspr. Kettenbuch. Kanten leicht abgeschrägt und gerade. Rücken: braunes Leder, drei Doppelbünde, von Goldbordüren gesäumt. Im ersten Feld Signaturschild (s. bei G), im zweiten Golddruck M. S. HASELBACH SERMONES in Goldrahmung. Kapitale mit Leder umstochen. Am Spiegel des VD Signaturen und Exlibris (s. bei G), am Spiegel des HD aufgeklebter beschriebener Pergamentfalz (s. bei Inhalt) und Tintenkleckse. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Bl. 1r Besitzvermerk 18. Jh. Collegii Neocell. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`1 F 10 (Bleistift) und Nr. 53 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 53. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. Ir Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 61, 282. – M. Stieglecker, Verborgene Zeichen. Handschriftenbeschreibung und Wasser zeichenanalyse, in: B. Sara (Hrsg.), Quelle & Deutung I. Beiträge der Tagung Quelle und Deutung I am 27. November 2013. Budapest 2014, 29–52, hier 44–48. – Dies., Hidden signs: how to capture watermarks, in: P. Ó Macháin (Hrsg.), Paper and the Paper Manuscript. A context for the transmission of the Gaelic litera ture. Cló Torna 2019, 44–54, hier 53. Thomas Ebendorfer (Haselbach): Sermones de tempore de evangeliis, pars hiemalis (Schneyer II Nr. 135–185. A. Lhotsky, Thomas Ebendorfer. Ein österreichischer Geschichtsschreiber, Theologe und Diplomat des 15. Jahrhunderts [Schriften der MGH 15]. Stuttgart 1975 [unveränderter ND der Ausgabe von 1957] 75, Nr. 38). Sermones jeweils zweigeteilt, Tit. und Inc. beider Teile, Expl. nur des gesamten Sermo aufgenommen. Die Schneyer-Nr. bezieht sich, falls nicht anders angegeben, auf den gesamten Sermo. (Ir–v) leer bis auf Stempel (s. bei G). 98 Cod. 53 (1r) am oberen Blattrand Tit.: Incipiunt sermones dominicales hyemales egregii doctoris magistri Thome Haselbach magistri arcium ac sacre pagine professoris et primo prima dominica in adventu domini etc. primus sermo prime dominice. Von späterer Hand 18. Jh.: Thomæ Haselbach. (1ra) Dnca 1. adv. (Schneyer II Nr. 135). Inc.: Cum appropinquasset Ihesus Ierosolimis et venisset Bethfage ad montem Oliveti tunc misit duos discipulos etc. Mt. 21 (1). Hodie sancta mater ecclesia incipit celebrare adventum domini et hanc observanciam recepit a sanctis patribus … – (1va) Tit.: Secundus sermo prime dominice. – Inc.: Dicite filie Syon Ecce rex tuus venit tibi manswetus etc. ubi supra (Mt 21,5) et originaliter Zacharie 9no capitulo (9). Secundum doctores adventus domini agitur per quattuor ebdomadas ad denotandum quod est quadruplex adventus filii Dei … – Expl.: … ut eternis solemnitatibus mereamur interesse Amen. (6va) Dnca 2. adv. Tit.: Dominica secunda in adventu domini. Sermo primus (Schneyer II Nr. 136). – Inc.: Erunt signa in sole luna et stellis in terra pressura Luc. 21o (25). Christus dominus per scripturas nobis valde incutit timorem de extremo iudicio … – (7ra) Tit.: Sermo (gestrichen: secundo) secunde dominice. – Inc.: Erunt signa in sole luna et stellis etc. Luc. 21o ubi supra (25). In presenti ewangelio monet nos dominus quatenus preparemus nos ad istam ultimam racionem quam oportet nos facere in extremo iudicio … – Expl.: … et mereamur intrare gaudia sanctorum Amen. – (11rb) Tit.: Sermo 3us 2e dominice (Schneyer II Nr. 137). – Inc.: Erunt signa in sole luna et stellis etc. Luc. 21o (25). Quod mul tum terribile sit extremum iudicium videtur posse trahi ex hiis que in proximo sermone dixi … – (13ra) Tit.: Sermo quartus 2e dominice. – Inc.: Arescentibus hominibus pre timore et expectacione que super venient universo orbi (Lc 21,26). In precedenti sermone dixi de signis adventus diei iudicii … – Expl.: … et tunc veniet dies domini scilicet iudicii de quo in sequenti sermone. – (19rb) Tit.: Sermo quintus 2e dominice (Schneyer II Nr. 138). – Inc.: Et tunc videbunt filium hominis venientem in nube cum potesta te magna et maiestate Luc. 22o (21,27). Sicut valde consultum est servo procuratori alicuius potentis domini qui numquam racionem fecit … – (20ra) Tit.: Sermo sextus secunde dominice. – Inc.: Et tunc videbunt filium hominis venientem in nube etc. ubi supra (Lc 21,27). In istis verbis describit salvator adventum suum ad iudicium extremum … – Expl.: … Ite maledicti etc. et e contrario de bonis Amen. (25vb) Dnca 3. adv. Tit.: Dominica 3a in adventu domini (gestrichen: Primus) sermo (Schneyer II Nr. 140). – Inc.: Cum audisset Iohannes in vinculis opera Christi mittens duos de discipulis suis ait illi etc. Mt. 11o (2). Ex quo inter cetera multum provocat ad virtutes et virtuosorum hominum laudabilis conversacio … – (26ra) Tit.: (gestrichen: Secundus) sermo tercie dominice in adventu domini. – Inc.: Cum audisset Iohannes in vinculis etc. ubi supra (Mt 11,2). Pro intellectu notandum quod secundum magistrum historiarum Iohannes arguebat Herodem propter Herodiadem quam a fratre suo recepe rat … – Expl.: … spinas et tribulos (korr. aus tribuualos) mundane cupiditatis extirpat (!). Miserunt Iudei ab Ierosolimis sacerdotes et levitas ad Iohannem ut interrogarent eum Tu quis es Ioh. 1o capitulo (19). (30ra) Dnca 4. adv. Tit.: Incipit prohemium quarte dominice (Schneyer II Nr. 141). – Inc.: Miserunt Iudei a Ierosolimis sacerdotes et levitas ad Iohannem ut interrogarent eum Tu quis es Iohannis 1o ca pitulo (19). (am Rand ergänzt: Et in ewangelio hodierno sicut scriptura) sacra Veteris et Novi Testa menti perswadet fideles ut diligant et amplexentur humilitatem … – (30va) Tit.: Dominica quarta sermo. – Inc.: Miserunt Iudei a Ierosolimis sacerdotes et levitas etc. ubi supra (Io 1,19). Secundum Originem Iudei videntes prope esse tempus adventus Messie … – Expl.: … et quam tardus ad bona et pronus ad mala etc. etc. (43rb) In nativitate dni. Tit. (am oberen Blattrand): In nocte nativitatis Christi. Sermo primus. – Inc.: Exiitt (!) edictum a Cesare Augusto ut describeretur universus orbis Luc. 2 (1). Sicut omnipotens Deus verax est in premissis Ita et misericors … – (43va) Inc.: Exiit edictum a Cesare Augusto ubi supra (Lc 2,1). Nota Octavianus imperator dictus est Augustus primus quia regnum Romanorum notabiliter auxit … – Expl.: … eciam quia stetit inter lapidantes. – (47va) Tit. (am oberen Blattrand): In nativitate domini nostri Ihesu Cristi. Sermo secundus. – Inc.: Exiit edictum a Cesare Augusto ut describeretur universus orbis Luc. 2 (1). Sicut dicit Bernardus in sermone de epyphania Priusquam appareret Dei Cod. 53 99 humanitas latebat … – (47vb) Inc.: Exiit edictum a Cesare Augusto (Lc 2,1). Secundum hystoriam post mortem Iulii Cesaris qui a poetis conmendatur … – Expl.: … ut cum angelis eternaliter sibi decante mus (gestrichen: post) laudes in patria Amen. (52rb) De s. Stephano. Tit.: Sequitur sermo in die s. Stephani (am oberen Blattrand: In aurora diei na tivitatis Christi. Alius sermo). – Inc.: Pastores loquebantur ad invicem Transeamus usque Betlehem et videamus hoc verbum quod factum est etc. Luc. 2 (15). Hodierne diei gaudiosa festivitas nos ammonet et accendit aliquid de temporali Christi … – (52vb) Tit.: In die s. Stephani. – Inc.: Pastores loquebantur etc. (Lc 2,15). Notandum quod Octavianus imperator 42 anno imperii sui quia 57 annis regnavit et versus finem valde pacifice … – Expl.: … ad Bethlehem celestem lapides torrentes contra eum gauden ter suscepit. (57va) Dnca infra octav. nativitatis dni (Schneyer II Nr. 142). – Inc.: Erat Ioseph et Maria mater Ihesu mirantes super hiis que dicebantur de illo etc. Luc. 2 (33). Augustinus in quodam sermone de Christi nativitate qui incipit Adiuvet nos humilimos … – (58rb) Inc.: Erat Ioseph et Maria mater Ihesu mirantes etc. (Lc 2,33). Ewangelium presens tangit primo de parentibus salvatoris id est patre et matre … – Expl.: … sed a casu similiter eveniunt etc. (63ra) In circumcisione dni. – Inc.: Postquam consumati sunt dies octo ut circumscideretur (!) puer vocatum est nomen eius Ihesus etc. Luc. 2o capitulo (21). Ex quo tota Christi vita et conversacio nostra est instruccio … – (63va) Inc.: Postquam consumati sunt dies octo ubi supra (Lc 2,21). Sciendum quod Deus creavit primos parentes iustos iusticia naturali id est sine omni obliquitate … – Expl.: … omnes filii Israhel incircumscisi (!) sunt corde etc. (66rb) In epiphania dni. Inc.: Cum natus esset Ihesus in Betlehem Iude (!) etc. Mt. 2o (1). Omnipotens Deus ex supereffluenti non ex operibus iusticie que fecimus … – Expl. abweichend: … discessimus ad hec parlamenta revocamur plura ibidem. – (70va) Tit. (am oberen Blattrand): In epiphania domini. Sermo secundus. – Inc.: [C]um natus esset Ihesus in Betlehem Iude (!) etc. Mti 2o (1). Sicut Deus dispo suit omnia sapienter sic fidei incrementum et suam humanitatem … – (71ra) Inc.: [C]um natus esset Ihesus vide supra (Mt 2,1). Pro intellectu littere notandum ut narrat Iosephus 13 libro Antiquitatum capitulo 14 regnum Iudeorum fuit interceptum 481 annis et duos menses a captivitate … – Expl.: … in aliis terris sunt eciam alia signa usque hodie mansura etc. (76ra) Dnca 1. post epiph. dni (Schneyer II Nr. 143). Inc.: Cum factus esset Ihesus annorum 12 etc. Luce 2o (42). Cristus dominus ne dum etate virili sed eciam in puericia sua se nobis ita exhibet ut ostendat quanta caritate nos diligat … – (76va) Inc.: Cum factus esset Ihesus annorum duodecim ascendentibus illis in Ierusalem secundum conswetudinem etc. (Lc 2,42). Augustinus dicit in De con sensu ewangelistarum et magistri hystoriarum quod Herode mortuo … – Expl.: … assiduat illi flagella ut letetur in novissimo suo et plura de hoc ibidem etc. – (80va) Tit.: Dominica de eodem sermo (Sch neyer II Nr. 144). – Inc.: Puer Ihesus proficiebat sapiencia etate et gracia apud Deum et homines. Luce 2o (52). Quando diligenter consideramus inestimabilia divine bonitatis beneficia nobis exhibita … – (81rb) Inc.: Puer Ihesus proficiebat etate etc. capitulo et loco ubi supra (Lc 2,52). Presens ewangelium tangit statum Christi tempore puericie circa 12 annum … – Expl.: … quibus parata sunt celestia per Christum dominum qui vivit et regnat in seculorum secula Amen etc. (84vb) Dnca 2. post epiph. dni. Tit. (am oberen Blattrand): Dominica secunda post epiphaniam. Sermo primus (Schneyer II Nr. 145). – Inc.: Nupcie facte sunt in Chana Galilee Iohannis 2o capitulo (1). Propter multiplicacionem humani generis Deus specialiter instituit matrimonium … – (85rb) Inc.: Nupcie facte sunt in Chana Galilee etc. ut supra (Io 2,1). Hic possit queri de tempore et loco et sic de aliis. Ad illud respondet ewangelista Iohannes 2o … – Expl.: … et allegat magister in quarta distinc cione versus finem. – (89vb) Tit. (am oberen Blattrand): Dominica secunda post epiphaniam. Sermo secundus. – Inc.: [N]upcie facte sunt in Chana Galilee Iohannis 2o (1). De prima parte ewangelii di cendum est sed referendo ad nupcias corporales de quibus loquitur specialiter ewangelium … – Expl.: … observare volens matrimonium contrahere vide Holkoth super Sapiencie etc. – (92rb) Tit. (am obe ren Blattrand): Dominica secunda post epiphaniam. Sermo tercius (Schneyer II Nr. 146). – Inc.: [N] upcie facte sunt in Chana Galilee etc. Iohannis 2o (1). Quamvis in nupciis homines soleant convivia 100 Cod. 53 tenere hec autem caute tenenda sunt in Dei timore … – (92vb) Inc.: [N]upcie facte sunt in Chana Galilee ut supra (Io 2,1). Prima pars presentis ewangelii congruit multum presenti tempore in quo celebrantur nupcie et matrimonia … – Expl.: … de tempore quo nupcie sunt celebrande vide supra etc. (96va) Dnca 3. post epiph. dni. Tit. (am oberen Blattrand): Dominica tercia post epiphaniam. Sermo primus (Schneyer II Nr. 148). – Inc.: [C]um autem descendisset Ihesus de monte secute sunt turbe multe. Et ecce leprosus etc. Mti octavo (1). Cristus dominus volens omnem dubitacionem de sua divini tate a nobis excludere … – (97ra) Inc.: [C]um autem Ihesus descendisset de monte etc. (Mt 8,1) Secun dum Chrysostomum omelia 24 super Mattheum Ne forte diceret populus aput se … – Expl.: … ut eterna premiati gloria ista mala divina gracia adiuvante vincamus Amen. – (110va) Tit.: Dominica 3a post epyphaniam (Schneyer II Nr. 147). – Inc.: Cum autem descendisset Ihesus de monte Ecce leprosus ve niens adorabat eum. Mt. 8us (1). Ecclesia instituit quod per totum annum cantantur et predicantur Christi miracula … – (110vb) Inc.: [C]um autem descendisset Ihesus de monte ubi supra Mt. 8vo capi tulo (1). Quia postillacio verborum ewangelii satis patet ex alio sermone facto de eodem. Ideo solum modo dicendum erit … – Expl.: … dicente domino Vade sicut credidisti fiat tibi adiuvante domino no stro Ihesu Christo benedicto in secula etc. (114va) Dnca 4. post epiph. dni (Schneyer II Nr. 149). Inc.: Ascendente Ihesu in naviculam secuti sunt eum discipuli eius etc. Mt. 8vo (23). Licet mundus iste primo appareat valde amicus tranquillus et paci ficus hominibus … – (115ra) Inc.: Ascendente Ihesu in naviculam etc. ubi supra Mt. 8vo capitulo (23). Cristus dominus tempore quo suam predicacionem incepit semper operatus est … – Expl. … hic place amus sibi in via ut secum mereamur congregari in patria etc. etc. (119va) Dnca 5. post epiph. dni (Schneyer II Nr. 151). Inc.: In illo tempore respondens Ihesus dixit Confiteor tibi pater domine celi et terre quia abscondisti hec a sapientibus et prudentibus etc. Mt. 11o (25). Dominus Deus non invitat solum cottidie per suos nunccios predicatores sed eciam per se ipsum … – (119vb) Inc.: Respondens Ihesus dixit Confitebor tibi, pater, domine celi et terre etc. Mt. 11 (25). Hic post postillacionem ewangelii alias habitam dicam solum de illa parte Venite ad me … – Expl.: … quo quisque Deo appropinquat hic in via per graciam et in eterna beatitudine per gloriam Amen. Sequitur. (124ra) Dnca in Septuages. (Schneyer II Nr. 152). Inc.: Simile est regnum celorum homini patrifamilias qui exiit mane conducere operarios in vineam suam etc. Mt. 20 c. (1). Dominus Deus creavit hominem (gestrichen: […]ro, am Rand ergänzt: ymo) et totum mundum propter suam bonitatem … – (124va) Inc.: Simile est regnum celorum homini patrifamilias etc. capitulo ubi supra (Mt 20,1). Chrysostomus super Mattheum omelia 28 Homo Christus paterfamilias cui celi et (gestrichen: fr) pater (recte: terra) una est domus familia … – Expl.: … pervenire mereamur ad requiem perpetuam prestante domino Ihesu Christo benedicto in secula seculorum etc. (129va) Tit.: Alius sermo in LXXma (Schneyer II Nr. 153). – Inc.: Simile est regnum homini patrifamilias etc. Mt. 20 (1). Sicut patrifamilias placet servus qui parate exequitur suam voluntatem … – (130ra) Inc.: Simile est regnum celorum homini patrifamilias qui exiit etc. capitulo ubi supra (Mt 20,1). Salva tor Christus in presenti ewangelio movet singulos homines ad considerandum sua propria opera … – Expl.: … ubi murmurabunt sine fine eis in maiorem confusionem et penam etc. (134ra) Dnca in Sexages. (Schneyer II Nr. 155). Inc.: Cum turba plurima conveniret et de civitatibus properarent ad Ihesum dixit per similitudinem Exiit qui seminat seminare semen suum etc. Luce 8 (4). Beatus Bernardus in sermone De Quadragesima dicit quod septem sunt impedimenta … – (134va) Inc.: Cum turba plurima conveniret convenirent (!) et de civitatibus properarent ad Ihesum etc. capitulo ubi supra (Lc 8,4). Presens ewangelium mea non indiget exposicione quia summa veritas … – Expl.: … eris potestatem habens supra decem civitates. – (138va) Tit.: Alius sermo in Sexagesima (Schneyer II Nr. 156). – Inc.: Cum turba plurima conveniret et de civitatibus properaret ad Ihesum dixit per simili tudinem Exiit qui seminat seminare semen suum. Luc. 8 (4). Vera (am Rand ergänzt: paciencia) quanta bona et quantas utilitates afferat homini possumus ex diversis scripturis sacris colligere … – (139ra) Inc.: Cum turba plurima etc. capitulo ubi supra (Lc 8,4). In presenti ewangelio dominus ostendit mul torum auditorum Christi devocionem … – Expl.: … per quod et reduci ad regnum promeruit 2 Regum Cod. 53 101 16. – (142rb) Tit.: Alius sermo in LXa (Schneyer II Nr. 154). – Inc.: Cum turba plurima conveniret, de civitatibus properarent ad Ihesum dixit per similitudinem Exiit qui seminat seminare semen suum etc. Luc. 8 (4). Salvator noster Christus dominus sepius verbis suis nos amonet (!) non solum mala fuge re … – (142vb) Inc.: Cum turba plurima ubi supra (Lc 8,4). Secundum Bedam Satorem istum nullum melius quam Dei filium intelligere possumus … – Expl.: … per levam autem vita presens designatur per dexteram vere futura (am Rand ergänzt: ut dicit Gregorius). (145vb) Dnca in Quinquages. (Schneyer II Nr. 157). Inc.: Assumpsit Ihesus duodecim discipulos suos et ait illis Ecce ascendimus Ierosolimam et consumabuntur (!) omnia etc. Luce 18 (31). Celestis medicus quamvis vocatus ab infirmo homine propter originale peccatum … – (146rb) Tit.: Sermo in Quinqua gesima. – Inc.: Assumpsit Ihesus duodecim discipulos et ait illis Ecce ascendimus Ierosolimam etc. Luce 18 (31). In presenti ewangelio ponuntur duo principalia. Primum de Christi passione … – Expl.: … et tecum finaliter permanere in patria prestante eodem domino nostro Ihesu Christo benedictoin secula Amen. – (151va) Tit.: In Quinquagesima (Schneyer II Nr. 158). – Inc.: Assumpsit Ihesus duodecim discipulos suos et ait illis Ecce ascendimus Ierosolimam et consumabuntur omnia que scripta sunt per prophetas etc. Luce 18 (31). Secundum scripturas sanctorum homo per peccatum fit instabilis et mutabilis et despectus … – (152ra) Inc.: Assumpsit Ihesus duodecim discipulos suos et ait illis Ecce ascendimus etc. (Lc 18,31). Hoc ewangelium facit memoriam de Christi passione et secundum Wilhel mum Duranti … – Expl.: … Respice fides tua te salvum fecit etc. Quam salutem. (156rb) Dnca 5. post epiph. dni. Tit. (am oberen Blattrand): Dominica quinta post epiphaniam domini. Sermo secundus (Schneyer II Nr. 1501). – Inc.: Confiteor tibi pater domine celi et terre etc. Mt. 11mo (25). Sicut inter virtutes morales que pertinentes sunt ad mores exteriores precipue precipua est gra titudo … – (156vb) Inc.: Confiteor tibi etc. (Mt 11,25). Quia Christus multa signa fecit in civitatibus Iudeorum ipsi nichilominus in sua pertinacia permanserunt … – Expl.: … ipsum amoris brachio sustentabat et ideo ipsum non onerabat. (159ra) Dnca 1. in Quadrages. Tit.: Dominica prima in Quadragesima (am Rand ergänzt: Sermo pri mus) (Schneyer II Nr. 159). – Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo etc. Mt. 4 (1). Ex quo omnia tempora tempus (am Rand ergänzt: suum) habent ut dicit sapiens Ecclesiastes 3o Est tempus flendi … – (159vb) Tit.: Dominica Invocavit. – Inc.: Ductus est Ihesus in desertum ubi supra (Mt 4,1). Ex quo sequitur assumpsit eum dyabolus etc. Dubitatur a quo spiritu ductus sit in desertum … – Expl.: … sed facit eciam cum temptacione proventum de quibus patebit in loco suo etc. – (164ra) Tit.: Alius sermo in dominica prima XLme (am Rand ergänzt: Sermo 2us) (Schneyer II Nr. 161). – Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo etc. Mt. 4 (1). Totam vitam nostram plenam fecit Deus temptacionibus et eciam racionabiliter Deus permittit electos suos temptari … – (164va) Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo (Mt 4,1). Glossa dicit Christus non permisit se temptari nisi post baptismum … – Expl.: … et Gregorius dicit Illos iure pulsare negligit (!) quos iure possidere quieto se sentit. – (169rb) Tit.: Alius sermo in prima dominica XLme (am Rand ergänzt: Sermo tercius) (Schneyer II Nr. 162). – Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo Mt. 4 (1). Sicut in bello corporali securius pugnatur et cicius obtinetur triumphus contra hostes … – (170ra) Inc.: Ductus est Ihesus in desertum etc. ubi supra (Mt 4,1). Tota summa huius ewangelii est quod Christus voluntarie ivit ad desertum locum … – Expl.: … quibus ipsum vincere poterimus prestante domino nostro Ihesu Christo Amen. – (174ra) Tit.: Dominica prima in XLma (am Rand ergänzt: Sermo quartus) (Schneyer II Nr. 160). – Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo Mt. 4 (1). Sicut dicit Gregorius omelia 2a super ewangelia Miracula domini et salvatoris nostri sic accipienda sunt … – (174va) Inc.: Ductus est Ihesus in desertum a spiritu ut temptaretur a dyabolo etc. ubi supra (Mt 4,1). Pro intellectu dubitatur quale fuit illud desertum ad litteram … – Expl.: … apparet ex textu littere quod sic quia dicitur Tunc a ssumpsit eum. – (176vb) Tit.: Alius sermo de ieiunio et utilitate eius (Schneyer II Nr. 163. Lhotsky 83, Nr. 103). – Inc.: Et cum ieiunasset 40 diebus et 40 noctibus postea esuriit Mt. 4 (2). Sancti patres primitivi Christiane ecclesie instituerunt ieiunium fidelibus observandum … – Expl.: … quia non potuerunt sed quia non voluerunt etc. Sequitur. – (185va) Tit. (am oberen Blattrand): Alius sermo de ieiunio 102 Cod. 53 ( Lhotsky 83, Nr. 104). – Inc.: Crisostomus omelia 16a super Mt. dicit Omnium malorum tria et forciora sunt tria mala … – Expl.: … et fratres in tota medietate claustri non quiescere permitteret. – (191ra) Tit. (am oberen Blattrand): De ieunio sermo tercius (Lhotsky 83, Nr. 105). – Inc.: Nunc igitur de gula videndum est isto tempore sacro quia (gestrichen: quod) secundum Gregorium … – Expl.: … quod iam modus est (gestrichen: esse, am Rand ergänzt: semper est peccatum) peccatum etc. – (197vb) Tit. (am oberen Blattrand): Alius bonus sermo. De confessione et penitencia et consimilibus (Lhotsky 84, Nr. 107). – Inc.: Quia ut ait Ambrosius in sermone de 40ma Ecce nunc tempus acceptabile est in quo confessio a morte animam liberat … – Expl.: … est magis de uno quam de alio conterendum hec de contricione breviter sint dicta etc. – (205ra) Tit. (am oberen Blattrand): De confessione et penitencia. Sermo secundus (Lhotsky 84, Nr. 108). – Inc.: Nunc ergo de confessione videndum est ad quam homo obligatur qui peccavit mortaliter ex Dei institucione et precepto Iohannis 20 (22) ubi confessionem instituit quando dixit apostolis Accipite spiritum sanctum … – Expl.: … ut prodigalitas avariciam ut vere impleatur illud abyssus abyssum invocat etc. De satisfaccione supra dictum est. – (209rb) Tit. (am oberen Blattrand): De rebus male acquisitis (Lhotsky 84, Nr. 109, in zwei Teile unterteilt). – Inc.: Videte ne furtivus sit reddite eum […] dominis suis quia non licet vobis edere ex furto aliquid aut contingere (Tb 2,13). Verba sunt beati Thobie ad vocem edi cuiusdam balantis caprarum … – (227ra) Inc.: Dicto de restitucione fienda dampnificatis in rebus exterioribus nunc restat dicere de restitucione fienda in bonis anime et corporis … – Expl.: … secundum arbitrium boni viri precipue ad intensas ad medicos etc. De restitucione si placet vide S. T. Scotum in scriptis magistri Nicolai de Dinckelspühel multa invenies si placet recurre illuc. – (227vb) Tit. (am oberen Blattrand): Sermo de penitencia (Lhotsky 84, Nr. 106, in zwei Teile unterteilt). – Inc.: Facite dignos fructus penitencie (Mt 3,8 und Lc 3,8). Ita hortatur nos predicator penitencie beatus Iohannes Baptista in deserto predicans baptismum penitencie Mt. 3 et Luce 3 quod specialiter nobis dicitur hoc sacratissimo ieiuniorum tempore legimus … – (232va) Inc.: Novissime audistis quomodo in hoc sacratissimo tempore matamur (!) ad penitenciam … – Expl.: … cum sacerdos dicit quidquid boni feceris sit tibi in remissionem peccatorum tuorum. (236vb) Dnca 2. in Quadrag. (Schneyer II Nr. 164). Inc.: Egressus Ihesus secessit in partes Tiri et Sydonis etc. Mt. 15 (21). Inter cetera que reconciliant peccatorem Deo postquam in peccatum mortale corruit est recognicio … – (237ra) Datierung in Rot: 1452. – (237rb) Tit.: Sermo in dominica Reminisce re. – Inc.: Egressus Ihesus secessit in partes Tiri et Sydonis Mt. 15 (21). Pro intellectu notandum sicut habetur Mt. 14 Postquam Christus transfretavit per mare Tyberiadis venit Capharnaum … – Expl.: … quia quod ipsi apostoli non impetraverunt et obtinuerunt ipsa impetrat. (238va) Dnca 3. in Quadrag. (Schneyer II Nr. 165). Inc.: Erat Ihesus eiciens demonium et illud erat mutum etc. Luc. 11mo (14). Secundum quod dicit Beda in omelia de Christi transfiguracione ut patet Mt. 17 Salvator humani generis Christus dominus … – (239ra) Tit.: Dominica tercia in XLma (am Rand ergänzt: Oculi). – Inc.: Erat Ihesus eiciens demonium etc. ubi supra (Lc 11,14). Demones enim frequen ter plus solito ante et circa Christi adventum obsidebant homines … – Expl.: … Tu solus nosci corda filiorum hominum etc. Et exposicionem textus sequentis in sermone (gestrichen: sequenti complebo) sequenti usque ad finem complebo. – (240ra) (Schneyer II Nr. 166). Inc.: Erat Ihesus eiciens demonium et illud erat mutum etc. Luce 11o (14). Quam detestabile sit vicium invidie et fugiendum patet propter multa mala … – (240va) Inc.: Erat Ihesus eiciens demonium ubi supra (Lc 11,14). Crisostomus et Theo philus dicunt et habetur Mt. 12 Ille demoniacus erat mutus … – Expl.: … corpore per carnis assump cionem et sic de aliis est. Idem Beda dicit in omelia. (242rb) Dnca 4. in Quadrag. (Schneyer II Nr. 167). Inc.: Abiit Ihesus trans mare Galilee quod est Tybe riadis et sequebatur eum multitudo magna etc. Iohannis 6 (1). Licet secundum Augustinum in quodam sermone ad fratres Non debeamus loqui sublimia … – (242vb) Inc.: Abiit Ihesus trans mare Galilee ubi supra (Io 6,1). In hoc ewangelio considerantur triplices persone scilicet Christus discipuli et turbe … – Expl.: … sobrie pie et caste vivendo celestis sapiencie mereamur dulcedine recreari Amen. (246ra) Dnca 1. in passione dni (Schneyer II Nr. 168). Inc.: Dixit Ihesus turbis Iudeorum et principibus sacerdotum Quis ex vobis arguet me de peccato si veritatem dico quare non creditis michi etc. Iohannis 8 (46). Quod quilibet homo et maxime Christianus debeat fugere lites et contenciones … – (246va) Cod. 53 103 Inc.: Quis ex vobis arguet me de peccato etc. ubi supra (Io 8,46). Christus dominus frequenter in suis sermonibus arguit Iudeis super peccatis suis … – Expl.: … sustinendi precipue verbales ut patet in presenti ewangelio etc. (249ra) Dnca 2. in passione dni (Schneyer II Nr. 169). Inc.: Cum appropinquasset Iesus Ierosolimis et venit Bethfage ad montem Oliveti etc. Mt. 21 (1). Consuetudo est regum terre ut quando volunt aliqua seriosa bellica exercere … – (249va) Inc.: Cum appropinquasset Iesus Ierosolimis etc. ubi supra (Mt 21,1). Pro intellectu littere notandum quod Iohannis 12 dicitur Quando Iudei iam ex consilio Cayphe … – Expl.: … quod 6ta die separabatur ante pascha in eadem ymolandum pro peccatis mundi. (251va) Fer. 5. in coena dni (Schneyer II Nr. 171). Inc.: Sic comedetis illum renes vestros accingetis calciamenta vestra habebitis in pedibus tenentes baculos manibus et comedetis festinanter etc. Exodi 12 (11). Hec verba licet ad litteram sint dicta de agno pascali cuius ymolacione … – (252ra) Inc.: Sic comedetis illum renes vestros accingetis calciamenta vestra habebitis in pedibus etc. ut supra (Ex. 12,11). Nunc igitur iuxta premissa sicut dixi ad contemplacionem … – Expl.: … qui habet peccatum mortale de quo non est contritus sufficienter etc. – (257vb) Tit.: Alius sermo in cena domini (Schneyer II Nr. 172). – Inc.: Hoc facite in meam commemoracionem 1o Cor. 11 (24). In hodierna epistola do minus noster Ihesus Cristus de hoc mundo per passionem suam acerbissimam … – (258rb) Inc.: Hoc facite in meam commemoracionem ubi supra (1 Cor 11,24). Sacramentum mei corporis sumite quo ad fideles et conficite quo ad presbiteros … – Expl.: … meritum fidei evacuetur si tamen instaret martirium sentiret. – (265ra) Tit.: Alius sermo in cena domini (Schneyer II Nr. 173). – Inc.: Ante diem festum pasche sciens Ihesus quia venit hora eius etc. Ioh. 13 (1). Crisostomus super Iohannem tractans illud Ioh. 10. Facta sunt encenia … – (265va) Inc.: Ante diem festum pasce sciens Ihesus quia venit hora eius etc. ubi supra (Io 13,1) Sicut auris non potest saciari auditu nec oculus … – Expl.: … ut te digne collaudare nunc et in eternum per interminabilia secula Amen. – (272rb) Tit.: Alius sermo de cena et passione Christi. – Inc.: Sciens Ihesus quia venit hora eius ut transeat ex hoc mundo ad patrem Ioh. 13 (1). Dominus Ihesus Christus et magister noster preciosum nostre redempcionis precium … – (272va) Inc.: Sciens Ihesus etc. (Io 13,1). Quia in tentivum (!) passionis Christi et occasio fuit prodicio … – Expl. erweitert (mit Innsbruck, ULB Tirol, Cod. 421, Bl. 372va übereinstimmend): … non studuerit condolere ut ait Gregorius … tibi appropinquare et tecum in gloria habitare concedere digneris Amen. (280rb) Fer. 6. in parasceve (Schneyer II Nr. 174). Inc.: Egressus Ihesus cum discipulis suis trans torrentem Cedron ubi erat ortus etc. Ioh. 18 (1). Sapiens Ecclesiastes 3 (1 und 4) dicit quod omnia tempora tempus habent. Est tempus flendi et tempus ridendi … – (281ra) Inc.: Igitur locuturus hodie nunc amplius de dira Christi morte et innocenti passione … – Expl. erweitert: … et omnibus diebus vite nostre in gloria habitare. Sed dubitaret forte aliquis Iterum salvator noster mori voluit … et celesti tuo patri in conspectum glorie tue me miserum commendare in te enim vere credo te confiteor unde tu domine etc. (299ra) Dnca resurrectionis dni (am Rand ergänzt: Sermo primus) (Schneyer II Nr. 175). Inc.: Maria Magdalene et Maria Iacobi et Salomee emerunt aromata ut venientes ungerent Ihesum. Mr 16 et ultimis (1). Presens Novi Testamenti precipua festivitas inter ceteras venerabilis toto orbe terrarum … – (299vb) Inc.: Maria (gestrichen: Iacobi, am Rand ergänzt: Magdalene) et Maria Iacobi et Salomee (Mc 16,1). Hee due fuerunt ultime sorores ex parte matris cum virgine gloriosa … – Expl.: … et impetravit orando coram ymaginem virginis. Vide Ieronimi in legenda ipsius etc. – (304vb) Tit. (am oberen Blattrand): In die sancto pasche. Sermo secundus (Schneyer II Nr. 176). – Inc.: [M]aria Magdalene etc. Marci ultimo (16,1). Divina scriptura istam habet proprietatem quod in una sui parte fideles ammonet lacrimari … – Expl. abweichend: … ad plenam pietatis graciam te adiuvante pervenire mereamur etc. – (310ra) Tit. (am oberen Blattrand): In die sancto pasche. Sermo tercius (Schneyer II Nr. 177). – Inc.: [S]urrexit non est hic Marci 16 (6). Quando dignitatem festorum tocius anni clarius inspicimus invenimus quod ipsa presens pascalis sollempnitas … – Expl.: … de eius utero quando natus fuit in mundo. Rogemus etc. – (313vb) Tit. (am oberen Blattrand): In die sancto pasche. Sermo quartus (Schneyer II Nr. 178). – Inc.: [E]xpurgate vetus fermentum etc. prime Corinthorum 5o (7). Hodiernam pascalem videlicet festivitatem inter ceteras festivitates Christi precipuam … – Expl.: … ab omni mortis impetu tuum defendas populum etc. 104 Cod. 53 (318rb) Dnca 1. post pascha. Tit. (am oberen Blattrand): In octava pasche. Sermo primus (Schneyer II Nr. 179). – Inc.: Accipite spiritum sanctum quorum remiseritis peccata Ioh. 20 (22). Salvator noster Christus dominus sicut omnia in sua passione ostendit ut declararet suam humanitatem … – Expl.: … quia tu Deus protector noster es et solus gloriosus super omnem terram etc. (322rb) Tit. (am oberen Blattrand): In octava pasche. Sermo secundus (Bibelzitat wie Schneyer II Nr. 180). – Inc.: Cum ergo sero factum esset die illa una sabatorum etc. Ioh. 20 capitulo (19). Hodie divina dies resurreccionis colitur in quo (!) perfecta requies designatur … – Expl.: … et vitam eternam possidere quam etc. (325ra) Dnca 2. post pascha (Schneyer II Nr. 181). Inc.: Ego sum pastor bonus. Bonus pastor animam suam dat pro ovibus suis Ioh. X (11). Licet multe sunt cause in particulari quare lites et turbaciones surgunt inter homines in mundo … – Expl.: … vide in 2o sermone post octavam Penthecostes scilicet Homo quidam erat dives. (328va) Dnca 3. post pascha (Schneyer II Nr. 182). Inc.: [M]odicum et iam non videbitis me etc. Ioh. 16 (16). Imminente hora qua Christus dominus voluit pro nobis crucis passionem et mortem pati … – Expl.: … quod numquam aufferetur ab eis quod nobis prestare dignetur etc. (333rb) Dnca 4. post pascha (Schneyer II Nr. 183). Inc.: Vado ad eum qui me misit Ioh. 16 capitulo (5). Divina disposicione et providencia ab eterno Deus piissimus disposuit postquam Iudaycus populus … – Expl.: … Nescitis, quia amicicia huius mundi inimica est Deo etc. (338rb) Dnca 5. post pascha (Schneyer II Nr. 184). Inc.: [A]men Amen dico vobis. Si quid pecieritis patrem in nomine meo dabit vobis Ioh. 16 (23). Quamvis multis modis homo serviat Deo et pro peccatis suis satisfaciat … – Expl.: … et hoc eciam post 6 menses factum est ut patet ibidem etc. (344rb) Dnca 6. post pascha (Schneyer II Nr. 185). Inc.: [C]um venerit paraclitus quem ego mittam vobis a patre etc. Ioh. 15 (26). Misericors pater qui est prope est omnibus invocantibus eum in veritate quia nos orphanos non relinquit … – Expl.: … ut oportet assistere coram eo etc. (346vb) Tit. (am oberen Blattrand): Sermo de sanctis infra pascha. – Inc.: [S]tabunt iusti in magna con stancia etc. Sap. 5 (1). Circa quem textum est dicendum an sancti in eterna beatitudine quam habebunt post resurreccionem sedebunt vel stabunt … – Expl.: … Et hec misericordia propria est Deo Iohannes dicimus in Collecta Omnipotens sempiterne Deus cui proprium misereri semper et parcere etc. Parallelüberlieferung: u. a. Innsbruck, ULB Tirol, Cod. 421, passim (Vorbesitzer Neustift), mit gering fügigen Abweichungen in der Anordnung. Gegenüber Cod. 53 fehlen die Sermones Bl. 156rb, 313vb, 322rb, 338rb, 344rb und 346vb. Zur Überlieferung s. Lhotsky 75. (348rb–vb) leer. (HDS) Aufgeklebtes Fragment einer Urkunde des Bischofs von Brixen an Johannes, scholasticus aus Isny (de Ysnina). Perg. ca. 35 × 280, am seitlichen Rand jeweils mit Leder des Einbands überklebt. Erhaltener Schriftraum ca. 20 × 250. Drei Zeilen. Urkundenschrift (Trecento II), Brixen 2. H. 14. Jh. Inc. mut.: ] episcopus Brixinensis dilecto nobis in Christo Iohanni scolastici (!) de Ysnina salutem in domino. Ecclesiam parrochialem sancti […]usina nostre diocesis ad collacionem nostram pleno … – Expl. mut.: … eius dimissionem tibi conferimus et de eadem providemus [ U. S. Cod. 76 105 Cod. 76 SAMMELHANDSCHRIFT Perg./Pap. I, 75, I Bl. 270 × 205. T. I: Neustift, Mitte 15. Jh., T. II: St. Dorothea (Wien) und Neustift, 6. Jz. 15. Jh. B: Zwei Teile: I (I, 1–16), II (17–74[75], I). Foliierungen: 1) Älteste Foliierung (vor Zusammenbinden der zwei Teile) am oberen rechten Blattrand, tw. beschnitten 1–56, beginnend mit Kapitel 1 der Statuten (Bl. 19r). 2) Zweite Foliierung am unteren rechten Blattrand, tw. beschnitten bzw. überklebt 1–13, begin nend mit dem Prolog der Statuten (Bl. 17r). 3) Dritte Foliierung (im Zuge des Zusammenbindens 1457?) am rechten Blattrand mittig 1–74, beginnend mit der Regula s. Augustini (Bl. 1r). 4) Jüngste Foliierung 19. Jh. durch die ULB Tirol am rechten oberen Blattrand (hier berücksichtigt), entsprechend der dritten Foliierung; Fehler in der Zählung: 71 doppelt angeführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, drei einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. REGUL. S. AUGU. CUM COME. in Gold rahmung, im dritten Signaturschild (s. bei G). Spiegel sowie Vor- und Nachsatzbl. Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (Vide Grundzettel), Spiegel des HD leer. G: Zumindest T. II der Hs. wurde von Nikolaus, Propst des Augustiner Chorherrenstiftes St. Dorothea in Wien, im Zuge seiner Visitation des Klosters Neustift zurückgelassen und dort wohl von seiner Hand mit Visi tationsinstrument (dat. Neustift, 18. Juli 1457) u. a. ergänzt. Abschrift vom Augustiner Chorherrn Petrus Ephensteiner 1728 in Neustift, Stiftsbibl., Cod. 29, S. 115–239. Hs. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am VDS alte Signaturen der ULB Tirol II`1 H 8 (Tinte, Bleistift, ältere radierte Signatur überschrieben), daneben N. 76/(2) (Tinte, Bleistift). Am Rücken gelbes Signaturschild der ULB Tirol 76. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedens vertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wretschko–Sprung 14–15. – Hermann 145, Nr. 162. – Sparber 227. – Kristeller I 439. – Neuhauser, Neustift 82. – Katalog ULB Tirol I 227, 282. – Peintner, Stiftsbibliothek 122. – Twenty-Fifth Saint Louis Conference on Manuscript Studies: Abstracts of Papers. Manuscripta 42 (1998) 139–157, hier 152. – H. Fasching, Die Wiener Konstitutionen für Regulierte Augustiner Chorherren in Österreich im 15. Jahrhundert (Beiträge zur Kirchengeschichte Niederösterreichs 16, Geschichtliche Beilagen zum St. Pöltner Diözesanarchiv 33). St. Pölten 2008, 87–94 (Sigle N). – Baroffio 324, Nr. 16709. – Rischpler 50. I (I, 1–16) B: Perg., vereinzelt löchrig, Bl. 16 genäht. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 2.IV16. Gegenbl. zum Vor satzbl. als Spiegel auf dem VD aufgeklebt. Bl. 11r neuzeitliches gelbes Papiersignakel der ULB Tirol. S: 1) Bl. 1r–10v: Schriftraum 180 × 90, von Tintenlinien begrenzt, zu 20 Zeilen. Kalligraphische Bastarda. Am Blattrand tw. Notizen von späterer Hand. 2) Bl. 11r–15v: Schriftraum 215 × 130, von Bleistiftlinien seitlich begrenzt, zu 25 Zeilen, tw. auf Bleistift linierung. Textualis. Am Blattrand bisweilen Notizen von späterer Hand. A: 1) Bl. 1r–10v: Rote Auszeichnungsstriche. Bl. 1r sechszeilige Fleuronnée-Initiale: blauer Buchstabenkörper mit ausgesparten Halbpalmetten, die mit feinem Tintenstrich und roter Kreuzschraffur herausgearbeitet sind; im Binnenfeld auf rotem Grund Ähre aus gestrichelten, spitzknospenartigen Halbpalmetten (an den Stiel zurückgeklappt); als Besatz Halbpalmetten, durch Zackenlinien verbunden und mit Doppelstrichen verziert. Von der Hand des Hauptflorators der Neustifter Gradualien (Graduale Neocellense I, dat. 1442; Graduale Neocellense II, zw. 1442 und 1446). 2) Bl. 11r–15v: Rote Auszeichnungsstriche, Unterstreichungen, Paragraphzeichen, Überschriften. Bl. 12r Zeigehand. Satzmajuskeln tw. cadellenartig verziert. Bl. 11r zweizeilige rote Lombarde mit blütenförmiger Aussparung, Binnenfeld rot ausgemalt. S. R. 106 1 2 Cod. 76 (Ir–v) leer. (1r–10v) Regula s. Augustini (PL 32, 1377–1384. Regula recepta = Ordo monasterii, Satz 1 + Praeceptum [CPL Nr. 1839b]. Vgl. L. Verheijen, La règle de saint Augustin 1 [Études augusti niennes 15,1]. Paris 1967, 417–437). (11r–15v) Quaestio sollemnis. Tit.: Questio solempnis cuiusdam doctoris theologie coram papa Eugenio et cardinalibus de re ligiosis quando professus mortaliter aut venialiter peccat. – Inc.: Utrum religiosus professus et divina precepta cum substancialibus regule conservans … – Expl.: … Quam nobis consequi con cedat Ihesus Christus benedictus Amen. (16r–v) leer. II (17–74[75], I) B: Bl. 17 Perg. (Falz zw. Bl. 27 und 28). Bl. 18–74 Pap., tw. an den Rändern verstärkt. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Inns bruck, ULB Tirol, Cod. 199 (dat. 1458/59); Klosterneuburg, Stiftsbibl., CCl 147 (dat. 1464). Lagen (un sicher): (III+5)27 + 2.VI51 + 455 + (V–2+2)65 + (III+3)73(74) + 174(75) + (I–1)I* (Nachsatzbl., Pap. 18. Jh). Bl. 17– 19, 26, 27, 52–57, 66–68, 74 eingefügte Einzelbl. Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Bl. 17r neuzeitliches gelbes Papiersignakel. Bl. 63v tw. mit Papierblatt (korr. Text?) überklebt. S: Bl. 17r–18v Schriftraum 225 × 130, von Blind- bzw. Tintenlinien begrenzt, zu 38 Zeilen, Bl. 19r–73v Schriftraum 215 × 115, zumeist von Tintenlinien begrenzt. Bastarda von drei Händen: 1) Bl. 17r–26r, Z. 2 und Z. 6–33, Bl. 27r–54v und Bl. 63v, Z. 23–Bl. 73v. Schreiber möglicherweise Nikolaus, Propst von St. Dorothea, in Neustift 1457 (Bl. 66r Datierung der abgeschriebenen Visitationsurkunde Neustift, 1457 Juli 18); 2) Bl. 26r, Z. 3–5 und Z. 34–Bl. 26v; 3) Bl. 55r–63v, Z. 22. Bl. 74r Nachtrag: Schriftraum 115 × 165. Antiquakursive, Mitte 17. Jh. (1653?). Am Blattrand bisweilen Notizen von späteren Händen, tw. dt. A: Rubrizierung wohl von einer Hand: Rote Auszeichnungsstriche, Unter- bzw. Durchstreichungen, Über schriften, ein- bzw. zweizeilige Lombarden, tw. mit roter Deckfarbe gefüllt oder einfach verziert. Bl. 71Ir Cadelle mit Profilkopf in Federzeichnung. Repräsentanten. 3 (17r–53v) sog. „Wiener Statuten“: Statuta canonicorum regularium sub episcopis Pataviensibus Georgio et Leonhardo (1389–1451) ordinata pro monasteriis BMV in Dürnstein et s. Dorothea (Wien) (Ed. Fasching 149–454). (17r) Prolog. Tit.: Incipit prologus in statuta canonicorum regularium. – Inc.: Cum ex primo et principali nostre regule precepto teneamur habere cor unum et animam unam … – Expl.: … et doctorum sigillis communita. (17v) Register. (19r) Haupttext, bestehend aus 59 Kapiteln. Tit. (cap. 1): De recepcione noviciorum et pena dancium aut recipiencium aliquid pro ingressu de absolucione a censuris et preparacione ad confessionem generalem et de rebus eorum. Capitulum primum. – Inc.: Quando aliquis se cupit ad communem vitam transferre … – Expl.: … ut in scrutinio facile reperiantur. Explicit. Die anlässlich der Visitation von 1457 eingeführten sog. „Wiener Statuten“ wurden im Augus tiner Chorherrenstift Neustift wahrscheinlich bis ins 18. Jh. verwendet. Wohl als anonyme Privat arbeit wurden sie in abgeänderter Form 1695 bei Paul Nikolaus Führer in Brixen gedruckt: „Statuta Religiosa, Canonicis Regularibus Lateranensibus Collegii Neocellensis S. Mariae Ad Gratias observanda“ (vgl. Fasching 100–101). Zur breiten hs. Überlieferung vgl. Fasching 52–53. Text wie: Herzogenburg, Stiftsbibl., Cod. 12 (Prov. Augustiner Chorherrenstift Dürnstein). Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 117–194, „Libellus primus“. Cod. 76 107 4 (53v–63v) Libellus officiorum (Ed. Fasching 457–542). (53v) Prolog. Tit.: Incipit prologus in libellum officiorum. – Inc.: Preterea quam divinum officium concernunt … – Expl.: … vel contemptum. (54r) Register. (54r) Haupttext, bestehend aus 16 Kapiteln. Tit.: Incipit libellus officiorum de prelato et eius conversacione. – Inc.: Prelatus monasterii qui tam verbo quam exemplo fratribus sibi commissis preesse debet … – Expl.: … pater noster cum totidem ave Maria. Zur breiten hs. Überlieferung vgl. Fasching 52–53, Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 194–208, „Libellus secundus“. 5 (63v–65v) De plebanis (Ed. Fasching 554–557). Tit.: Sequitur capitulum de plebanis quales debeant esse. – Inc.: Plebani quia post prelatum maioribus quam ceteri fratres sunt periculis expositi … – Expl.: … non valeat induci. Schlussschrift: Expliciunt statuta. Parallelüberlieferung: Herzogenburg, Stiftsbibl., Cod. 12 (Prov. Augustiner Chorherrenstift Dürnstein); Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 215–218, „Libellus secun dus“, cap. 17. 6 (66r–71v) Abschrift des Visitationsinstruments des Augustiner Chorherrenstiftes Neustift durch Nikolaus, Propst von St. Dorothea in Wien, der von Nicolaus Cusanus im Zuge der Melker Reform als Visitator nach Neustift entsandt wurde. Tit.: Sequitur carta in monasterio Novacellensi relicta per reverendissimum patrem dominum Nicolaum praepositum s. Dorothee Vienne in visitatione 1457. – Inc.: In nomine domini Amen. Splendor paterne glorie … Datierung: Datum in sepe dicto monasterio Novecelle die lune decimaoctava mensis Iulii anno domini millesimo quadringentesimo quinquagesimo septimo. Originale Pergamenturkunde (Neustift, Stiftsarchiv, VV 50) mit Wachssiegel des Propstes Niko laus von St. Dorothea an Hanfschnur, dat. Neustift, 1457 Juli 18, bestätigt vom Brixner Bischof Nikolaus Cusanus. Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 219–228. 7 (71Ir–72v) Abschrift einer Cedula für das Augustiner Chorherrenstift Neustift, verfasst von Nikolaus, Propst von St. Dorothea in Wien, im Rahmen seiner Visitation des Klosters Neustift 1457 (Ed. Fasching 686–689). Tit.: Sequitur copia cedule per dominum prepositum sancte Dorothee hic relicte. – Inc.: Sicut tactum est in carta quadam hoc in loco reperta … Originale Pergamenturkunde nicht mehr erhalten. Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 228–231. 8 (73r–v) Über die Pfründnerschaft im Kloster, dt. (Ed. Fasching 572–573). Tit.: De donatis. – Inc.: Wer begert zu ainem donatum aufgenommen werden, der sol vor wissen, daz er nit zu müssichait und rue … – Expl.: … nyemands icht darumb gepunden noch schuldig. Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 231–233. 9 (74r) Kapitelbeschluss von 1653, betreffend die Vigil des Augustinustages in Neustift und die Frage der Abstinenz. Inc.: Anno salutis MDCLIII septimo Kalendis Septembris ad maiorem Dei gloriam S. P. N. Au gustini honorem … Abschrift in: Neustift, Stiftsbibl., Cod. 29 (dat. 1728), S. 234. (74v) leer. (Ir–v) leer. 108 Cod. 77 Zu Nikolaus de Corona (Kronstadt, Siebenbürgen), Propst von St. Dorothea in Wien (1428– 1458), und der Visitation in Neustift vgl. u. a. F. Röhrig, Wien – St. Dorothea, in: F. Röhrig (Hrsg.), Die ehemaligen Stifte der Augustiner-Chorherren in Österreich und Südtirol (Öster reichisches Chorherrenbuch. Die Klöster der Augustiner-Chorherren in der ehemaligen Öster reichisch-ungarischen Monarchie 1). Klosterneuburg 2005, 664–665; Fasching 19–24. Zu seiner Visitationstätigkeit vgl. auch M. Niederkorn-Bruck, Die Melker Reform im Spiegel der Visitatio nen (MIÖG Erg.-Bd. 30). Wien 1994. C. S. Cod. 77 THOMAS DE AQUINO. PETRUS BLESENSIS Pap. I, 120, I Bl. 275 × 200. Basel, 1465. B: Pap. bes. am Beginn und am Schluss fleckig. Nachweise und Bilder der Wasserzeichen s. WZMA. Wasser zeichenbefund mit Datierung übereinstimmend. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 10.VI119(120) + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Lage 3 und 4 vertauscht (s. bei Inhalt). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 47 doppelt angeführt. Bl. 107 und 113 Reste von neuzeitlichen gelben Papiersignakeln der ULB Tirol. S: Schriftraum 240/245 × 155/160. Zwei Spalten, Bl. 1r–47Iv von Bleistift- bzw. Blindlinien gerahmt, Bl. 48r– 119v von feinen Tintenlinien gerahmt, zu 51–61 Zeilen. Flüchtige Bastarda, dat. 1465 (Bl. 106vb, 112vb [Basel, 9. September.], 119va [13. September]). Am Blattrand Zählung der einzelnen Quaestiones, tw. be schnitten. A: Rote Auszeichnungsstriche, Paragraphzeichen (ähnlich Zeilenanschlusszeichen) und Unterstreichungen. Bl. 1ra sechszeilige rote Initiale mit ausgesparter vierblättriger Blüte, Bl. 113ra fünfzeilige Initiale nicht ausgeführt. Bl. 27v–36v, 47v–47Iv, 50v–59r, 68v–71v, 74v–95v, 101v–107v, 108v–119v keine Rubrizie rung. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck OPERA THEOLOGICA in Goldrahmung, darunter Signaturschild (s. bei G). Schnitt rot gesprenkelt. Am Spiegel des VD Signaturen, Exlibris und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 K 9 (Bleistift) und Nr. 77/(1) (Tinte, Bleistift) sowie Bleistiftnotiz (Thomas de Aquino Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 77(1). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 227, 283. – Stefani 95. 1 (Ir–v) leer. (1ra–112vb) Traktat in enger Anlehnung an: Thomas de Aquino: Super librum quartum Senten tiarum (Vgl. Druck: u. a. GW M46385. Ed. Opera omnia XI. Paris 1847. – Glorieux Nr. 14f4. Grabmann 286–290. Shooner Nr. 1126, 10). Gegenüber Druck bisweilen abweichend und gekürzt, Reihenfolge der Quaestiunculae vereinzelt abgeändert, Expositiones textus am Ende der einzelnen Distinctiones nicht immer enthalten, Kommentare jeweils an die einzelnen Quaestiones und Quaestiunculae direkt anschließend. Lage 3 und 4 (Bl. 25–47I) im Zuge der Neubindung vertauscht, daher Forts. von Bl. 24vb (Dist. 9) auf Cod. 78 109 Bl. 37ra, Forts. von Bl. 47Ivb (Dist. 14) auf Bl. 25ra, Forts. von Bl. 36vb (Dist. 16) auf Bl. 48ra (kein Textverlust). (1ra–106vb) Text. Inc.: Circa distinccionem sacramenti (!) queritur utrum hec distinccio sit bona. Sacramentum est invisibilis gracie visibilis forma ut ymaginem gerat et causa existat. Resspondo (!) quod sic quia completissime racionem sacramenti designat … (Dist. 1, qu. 1, art. 1, quaestiunc. 3) … Queritur utrum hec sit bona. Sacramentum est sacre rei signum respondeo quod sic quia sacramentum secundum proprietatem vocabuli videtur … (Dist. 1, qu. 1, art. 1, quaestiunc. 1) … – Expl.: … de quibus penis nos liberet et ad gaudium nos perducat qui sine fine regnat Amen. Enthält: (1ra) Dist. 1. – (4vb) Dist. 2. – (6vb) Dist. 3. – (8va) Dist. 4. – (11va) Dist. 5. – (13ra) Dist. 6. – (15rb) Dist. 7. – (17ra) Dist. 8. – (23ra) Dist. 9. – (25ra) Forts. Dist. 14. – (28rb) Dist. 15. – (36rb) Dist. 16. – (37ra) Forts. Dist. 9. – (38ra) Dist. 10. – (39va) Dist. 11. – (43ra) Dist. 12. – (46rb) Dist. 13. – (47Ivb) Dist. 14. – (48ra) Forts. Dist. 16. – (49vb) Dist. 17. – (54vb) Dist. 18. – (57va) Dist. 19. – (59ra) Dist. 20. – (60vb) Dist. 21. – (62rb) Dist. 22. – (63va) Dist. 23. – (64vb) Dist. 24. – (66vb) Dist. 25. – (68ra) Dist. 26. – (68va) Dist. 27. – (69va) Dist. 28. – (69vb) Dist. 29. – (70ra) Dist. 30. – (70va) Dist. 31. – (71ra) Dist. 32. – (71va) Dist. 33. – (73rb) Dist. 34. – (74rb) Dist. 35. – (74va) Dist. 36. – (75ra) Dist. 37. – (75rb) Dist. 38. – (77vb) Dist. 39. – (78ra) Dist. 40. – (79ra) Dist. 41. – (80ra) Dist. 42. – (80vb) Dist. 43. – (82rb) Dist. 44. – (88ra) Dist. 45. – (90va) Dist. 46. – (92vb) Dist. 47. – (94vb) Dist. 48. – (95vb) Dist. 49. – (104va) Dist. 50. (106vb) Schlussschrift: Et sic est finis Questionum fratris Thome de Aquino ordinis Predicato rum super quartum Sentenciarum anno domini 1465. (106vb–112vb) Register, geordnet nach Distinctiones und Quaestiones. (112vb) Schlussschrift: Expliciunt Questiones super quartum Sentenciarum cum earum registro magistri Thome de Aquino cum solis solucionibus dimissis de gerentibus. In Basilea anno domi ni MCCCCoLXV (gestrichen: in die) secunda feria in die Kunigundis (9. September 1465). 2 (113ra–119va) Petrus Blesensis: Compendium in Iob (PL 207, 795C–826B. – Stegmüller RB Nr. 6431). Dem Kommentar ist ein Widmungsbrief an König Heinrich II. Plantagenêt vorange stellt. Schlussschrift: Finitum est Idus Septembris anno domini LXVo. Deo gracias (13. September 1465). (119vb) nur Rahmung. (Ir–v) leer. A. P. / U. S. Cod. 78 PEREGRINUS OPPOLIENSIS Pap. I, 120, I Bl. 280 × 205. Raum Brixen (Neustift?), um 1430/33. B: Pap. etwas fleckig, Bl. 119v stark verschmutzt. Bilder und Nachweise der Wasserzeichen s. WZMA. Dat ierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Neustift, Stiftsbibl., Cod. 177; Wilten, Stiftsbibl., Cod. 32 02 05, T. IV (dat. 1431); Innsbruck, ULB Tirol, Cod. 437, T. II (Stams, dat. 1432), Cod. 666, T. III (Schnals, dat. 1433). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 10.VI119(120) + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am oberen Blatt rand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 115 doppelt angeführt. S: Schriftraum 185/200 × 120/130. Zwei Spalten, von feinen Tintenlinien gerahmt, zu 35–43 Zeilen. Zirkel stiche. Bastarda von zwei Händen: 1) Bl. 1ra–60vb; 2) Bl. 61ra–119rb. Oberlängen der Buchstaben der ersten Zeile bisweilen gelängt. Vereinzelt Randnotizen und Kapitelzählung von späterer Hand. 110 Cod. 78 A: Rote Auszeichnungsstriche, Unterstreichungen, Überschriften und Schlussschrift. Rubrizierung (insbes. Titel) bisweilen nicht ausgeführt. Zeigehände. Zwei- bzw. dreizeilige rote Lombarden, tw. mit Punktverdi ckungen, tw. mit einfach verzierten Ausläufern. Ab Bl. 61 Repräsentanten. Bl. 1ra sechszeilige rote Lombar de mit leicht verziertem Ausläufer. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, mit Goldbordüren gerahmt. Im zweiten Feld Golddruck M. S. DE TEMP. 1460 (Datierung?) in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gefärbt. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 119v Besitzvermerk 15. Jh. Iste liber est monasterii s. Marie ad Novacellam. 1809 anlässlich der vo rübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signa turen der ULB Tirol II 1 K 10 (Bleistift) und Nr. 78 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 78. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 227, 283. (Ir–v) leer. (1ra–119rb) ‹Peregrinus Oppoliensis›: Sermones de sanctis (Schneyer 4, 558–572). Tit.: Incipiunt sermones de sanctis per circulum anni. (1ra) De s. Andrea (Nr. 195). (1vb) Nr. 196. (3ra) Nr. 197. – (4rb) De s. Nicolao (Nr. 198). Inc.: Bene diccionem omnium gencium dedit illi. Sap. (Sir 44,25). In verbis istis ostendit nobis spiritus sanctus quedam signa in beato Nicolao propter que signa obtinuit benediccionem omnium gencium … (4vb) Nr. 199. (6rb) Nr. 200. – (7ra) De s. Lucia (Nr. 201). – (7vb) De s. Thoma ap. (Nr. 202). Expl.: … in castitate Deo spiritum reddiderunt. (8vb) Inc.: Affer manum tuam et mitte in latus meum (Io 20,27). Benigni homines habent modum dum vident aliquem ex caris amicis suis cecidisse in aliquam passionem solent eos instruere … – Expl.: … sine mortalibus peccatis existentibus. Parallelüberlieferung: Neustift, Stiftsbibl., Cod. 617, Bl. 136r (Entstehungsort: Neustift, 1455); Inns bruck, ULB Tirol, Cod. 71, Bl. 21rb (Vorbesitzer: Wilten, Entstehungsort: Tirol, 15. Jh.), Cod. 207, Bl. 5va (Vorbesitzer: Schnals, Entstehungsort: Österreich, dat. 1440, Schreiber: Christianus de Engel precz). (10va) De s. Stephano (Nr. 203). (11rb) Nr. 204. (12ra) Nr. 205. Expl.: … venit ad amplexus domini (wie Innsbruck, ULB Tirol, Cod. 207, Bl. 7rb). (12va) Nr. 111 (Expl. auch Nr. 205, wie Innsbruck, ULB Tirol, Cod. 207, Bl. 7va). – (13vb) De s. Iohanne ap. (Nr. 113). Expl.: … recedente autem luce fovea plena manna reperitur sicut plenius supra dictum est (Expl. vgl. Innsbruck, ULB Tirol, Cod. 71, Bl. 26vb und Cod. 207, Bl. 8ra). (15ra) Nr. 206. (15vb) Nr. 207. Expl. (erweitert): … sed perseveravit in fide Christi ut vicissitudinem Christi amicicie proposse redderet etc. (Expl. wie Innsbruck, ULB Tirol, Cod. 71, Bl. 29va und Cod. 207, Bl. 9va). – (16va) De s. Innocentibus (Nr. 208). (17rb) Nr. 209. Expl.: … carnifices guttura eorum gladio figebant ridebant. Sic et tu 2m et 3m collige supra etc. (vgl. Innsbruck, ULB Tirol, Cod. 71, Bl. 31rb und Cod. 207, Bl. 10rb). (18ra) Nr. 210. Expl.: … pro eo quod non correxit eum a iuventute sua (Expl. vgl. Innsbruck, ULB Tirol, Cod. 207, Bl. 10va). (18rb) De s. Thoma martyre (Nr. 211). – (19rb) De s. Fabiano et Sebastiano (Nr. 212). – (19vb) De s. Agnete (Nr. 213). – (20vb) In conversione s. Pauli (Nr. 214). (22rb) Nr. 215. (23va) Nr. 216. Expl.: … quod lac fluxit pro sangwine de corpore suo (Expl. vgl. Innsbruck, ULB Tirol, Cod. 71, Bl. 39vb und Cod. 207, Bl. 15ra). – (25rb) In purificatione BMV (Nr. 217). (26rb) Nr. 218. (27va) Nr. 219. (29rb) Tit.: Idem sermo. – Inc.: Ecce ego mitto angelum meum qui preparabit viam ante faciem tuam. Malach. 3o (1). In verbis istis 4or (Innsbruck, ULB Tirol, Cod. 207, Bl. 18ra: quinque) notantur. Primum est quod dominus dicit Ecce ego mitto angelum meum etc. 2m est quod dicit … – Expl.: … ut nos cum illo et ipse nobiscum usque in finem perseveret. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 71, Bl. 45va und Cod. 207, Bl. 18ra. Cod. 78 111 (30va) In cathedra s. Petri (Nr. 220). (31va) Nr. 221. (32vb) Nr. 124. Expl.: … ipsum imitari. Exaltavit et eum in virtute miraculorum. Hic dic miracula. – (33ra) De s. Matthia (Nr. 222). (35ra) (Bibelzitat und Expl. wie Nr. 223). Inc.: Tu domine qui nosti corda hominum etc. Actus (1,24). Hodie in ecclesia agitur de duabus personis scilicet de Iuda qui est de honore suo detrusus … (36ra) Nr. 224. – (37ra) De s. Gregorio (Nr. 126). Expl.: … semper sum circa te. Ecce quanta felicitas (Expl. wie Innsbruck, ULB Tirol, Cod. 207, Bl. 22vb). (38ra) Nr. 225. – (38vb) De s. Benedicto (Nr. 127). Expl.: … dominus parvulos amplexatus est et eos ad se vocari facit prosequere sicut vis (vgl. Expl. Nr. 226). – (39vb) In annuntiatione BMV (Nr. 227). (42ra) Nr. 228. (43va) Nr. 229. (44vb) Nr. 230. Expl.: … Hic dic exem plum de muliere et puero require supra. (45ra) Nr. 232. Expl.: … ut mediante gracia sua perveniamus ad gaudia eterna etc. (Expl. wie Innsbruck, ULB Tirol, Cod. 207, Bl. 27rb). (46vb) Nr. 233. – (48vb) De s. Ambrosio (Nr. 129). Expl.: … quid sic factum est et sic patet quomodo exaltatus est in presenti (Expl. wie Innsbruck, ULB Tirol, Cod. 207, Bl. 29rb). – (49vb) De s. Georgio (Nr. 234). (51va) Nr. 235. – (52vb) De s. Marco (Nr. 132). Expl.: … quia potest nos iuvare corpore et anima (Expl. wie Innsbruck, ULB Tirol, Cod. 207, Bl. 30vb). (53vb) Nr. 236. – (55rb) De s. Philippo et Iacobo (Nr. 134). Expl.: … peccatores in magna constancia. (56ra) Nr. 238. – (57va) In inventione s. crucis (Nr. 239). Expl.: … vestitus regia veste. Sic etc. (Expl. wie Innsbruck, ULB Tirol, Cod. 207, Bl. 32rb). (58vb) Nr. 136. (60ra) Nr. 240. – (61rb) De s. Iohanne ante Portam Latinam (Nr. 241). (61vb) Nr. 242. Expl.: … ubi pax ibi Deus et ubi Deus ibi omne bonum etc. – (62rb) De s. Barnaba (Nr. 140). Expl.: … et eum ibidem protinus combusserunt. Et sic amiciciam Dei est consecutus etc. – (63rb) De s. Vito (Nr. 243). Expl.: … ne per accidiam separaretur ab eo. – (64ra) De s. Iohanne Baptista (Nr. 142). Expl.: … totus mundus est consolatus. (64vb) Tit.: De eodem. – Inc.: Tu puer propheta altissimi vocaberis preibis enim ante faciem domini parare vias eius (Lc 1,76). Nota quod est triplex via. Prima in mundum, 2a per mundum, 3a extra mundum. Prima via in mundo (!) est lamentabilis … – Expl.: … Propterea circumdatus es laqueis etc. (65va) De s. Petro et Paulo (Nr. 248). Expl.: … in omni vita sua nichil aliud operatus est nisi quod laboravit oravit et predicavit. – (67ra) De s. Margareta (Nr. 252). – (68ra) De s. Maria Magdalena (Nr. 253). (69ra) Nr. 255. (69vb) Ohne Tit. Inc.: Ponam arcum meum in nubibus celi et erit signum federis mei. Gen. 9 (13). Hec verba possunt exponi de beata Maria Magdalena que intelligitur per arcum celestem propter tria. Primo racione materie … – Expl.: … quantum optimam partem celestis glorie elegit dicitur illuminata (vgl. ‹Aldobrandinus de Cavalcantibus› [= ‹Ps.-Thomas de Aquino›]: Sermo 53: De s. Maria Magda lena [Druck: S. E. Fretté, Thomae Aquinatis Opera Omnia 29. Parisiis 1876, 317]. – Vgl. Schneyer 8, 620, Nr. 36). (70ra) De s. Iacobo (Nr. 257). Expl.: … potestis calicem bibere. Responderunt audacter Possumus. (71ra) Nr. 259. – (72ra) De s. Petro ad vincula (Nr. 149). Expl.: … et regni eius non erit finis. (73ra) Nr. 262. – (73va) De s. Laurentio (Nr. 263). Expl.: … et hoc spontanee quia voluntarie ibat ad supplicia dic legendam. (74ra) Nr. 264. (75ra) Nr. 150. Expl.: … Imperatorem tunc obisse et unam aurem fractam esse in testimonium. – (75vb) In assumptione BMV (Nr. 151). Expl.: … et multis retulit que vidit et audivit et sic multi crediderunt etc. (77rb) Nr. 265. (78va) Nr. 266. (79va) Nr. 267. (80ra) Ohne Tit. Inc.: Vidi alterum angelum descendentem de celo amictum nube et yris in capite eius et facies eius sicut sol Apoc. (10,1). In verbis istis tria notantur de sancto (gestrichen: Barth, am Rand korr. zu: Bernhardo). Primo sanctitas vite cum dicitur Vidi alterum angelum descendentem de celo amictum nube. Primus enim angelus fuit beatus Benedictus … – Expl.: … in peccatis vivificavit suis miraculis sicut in vita sua legitur etc. (80vb) De s. Bartholomaeo (Nr. 268). (81rb) Nr. 269. Expl.: … quattuor seris scilicet pudore con fitendi spe longioris vite timore pene et desperacione etc. (82vb) Nr. 270. (83vb) Nr. 271. (85ra) De s. Augustino (Nr. 272). – (86rb) In decollatione s. Iohannis Baptistae (Nr. 273). (87rb) Nr. 274. Expl.: … dyabolus decollavit eum etc. – (87vb) In nativitate BMV (Nr. 275). (88vb) Nr. 276. (89vb) Nr. 155. Expl.: … et cognoscamus et amemus eum ut ab ipso numquam seperemur (!) etc. (91rb) Nr. 277. Inc.: Nova lux oriri visa est honor et tripudium aput omnes populos urbis (!) et provincias Hester Vo (recte: 112 Cod. 78 8,16–17). Quia nativitatem beate Marie agimus ideo gaudere debemus quia ipsa est stella … – (92vb) In exaltatione s. crucis (Nr. 278). (94ra) Nr. 156. Expl.: … ut te eriperet de faucibus leonum infernalium id est demonium. – (95rb) De s. Matthaeo (Nr. 279). (96va) Nr. 280. (97vb) Tit.: De sancto Mauricio. – Inc.: Absterget Deus omnem lacrimam etc. Apoc. (7,17 und 21,4). Ad mimitacionem (!) sanctorum flere debemus et nos quinque de causis … – Expl.: … Lacrimis meis stratum meum rigabam etc. (98va) De s. Michaele et de s. angelis (Nr. 284). Inc.: Omnes sunt aministratorii spiritus in ministerium missi propter eos qui hereditatem capiunt salutis gloria id est hominis quibus promittitur vita eterna Heb. II (recte: 1,14). Modus est nobilium et aliorum honestorum virorum quod ad filiorum suorum custodiam et instruccionem ponunt suos servos … (99va) Nr. 285. (100va) Inc.: Omnes sunt aministratorii spiritus etc. Heb. IIo (recte: 1,14). Hodie agimus festum omnium angelorum et hoc propter plures causas. Primo quia singulariter omnes honorare non possumus. Isto die omnes honoremus … – Expl.: … et inter choros martirum collocavit etc. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 207, Bl. 63va. (101va) De s. Michaele et de s. angelis (Nr. 286). (103rb) Nr. 287. – (104rb) De s. Hieronymo (Nr. 288). – (104vb) De s. Luca (Nr. 289). Expl.: … et mortuos sedebit sicut ipse dominus ait In generacione etc. (Expl. vgl. Innsbruck, ULB Tirol, Cod. 207, Bl. 66ra). – (105vb) De s. Ursula et de undecim milibus virginum (Nr. 290). – (106va) De s. Simone et Iuda (Nr. 291). (107rb) Nr. 292. (108ra) Nr. 293. – (108vb) Tit.: De eodem (Schneyer: De omnibus Sanctis, Nr. 294). (109va) De omnibus Sanctis (Nr. 295). (110ra) Nr. 296. – (110vb) In commemoratione omnium fidelium defunctorum (Nr. 298). (111va) Nr. 299. (112ra) Ohne Tit. Inc. vgl. ‹Sifridus Niger› (Schneyer 5, 432, Nr. 49) bzw. anonymer Sermo (Schneyer 9, 535, Nr. 59, zu Stuttgart, LB, theol. fol. 332). Inc.: Mittite partes eis qui non preparaverunt sibi. (2) Esdre (8,10). Consuetudo in mundo est quando aliquis usque in mortem in carcere cruciatur mittit ad amicos suos de quibus maxime confidit … – Expl.: … illis qui sunt in purgatorio. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 207, Bl. 75rb. (113rb) Ohne Tit. Inc.: Miseremini mei etc. Iob 19 (21). Notandum monere penitencie portando debe mus fratres nostros exemplo Christi qui penam peccatorum portavit … – Expl.: … valuerunt oraciones omnium adiutorum pro eis facere etc. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 207, Bl. 76rb. (113va) In commemoratione omnium fidelium defunctorum (Nr. 300). Expl.: … Qui coronat te in mi sericordia etc. – (114ra) De s. Martino (Nr. 301). (115ra) Nr. 174. Expl.: … martir sanguine fuit tamen voluntate. (115Ira) Nr. 302. – (115Iva) De s. Elisabetha (Nr. 303). (116rb) Nr. 304. – (117ra) De s. Clemente (Nr. 305). – (117va) De s. Catharina (Nr. 306). (118va) Nr. 307. Expl.: … ut quicumque in memoria mei scriptam passionem meam habuerit etc. Rogemus. (119rb) Schlussschrift: Deo gracias. (119va–b) nur Rahmung und Besitzvermerk (s. bei G). (Ir–v) leer. U. S. Cod. 83 113 Cod. 83 GREGORIUS MAGNUS Pap. I, 59, I Bl. 285 × 205. Süddeutscher Raum, 1400. B: Bl. 1 beinahe lose. Buchblock nachträglich beschnitten. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Identische Wasserzeichen: Neustift, Stiftsbibl., Cod. 158 und Cod. 183. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + (V–1)9 + 2.VI33 + VII47 + VI59 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 1 herausgerissen (ohne Textverlust). Am Ende der Lagen 2–4 Reklamanten, tw. stark be schnitten. Am oberen Blattrand Foliierung 15. Jh. 70–128, Hinweis auf ehem. Sammelhandschrift (s. bei G). Fehler in der Zählung: 77 statt 70 (wohl Fehler in der Schreibung). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol (hier berücksichtigt). Bl. 52 Rest eines neuzeitlichen gelben Papiersignakels der ULB Tirol. S: Schriftraum 215 × 140/150. Zwei Spalten, von feinen Tintenlinien gerahmt, zu 34–49 Zeilen. Zirkelstiche. Bastarda von zwei Händen: 1) Bl. 1ra–52rb, dat. 1400 (Bl. 52rb), mit unterschiedlichem Duktus (= Schrei ber Cod. 158); 2) Bl. 52va–55vb (wohl von gleicher Hand wie Foliierung und Besitzvermerk Bl. 56r). A: Rote Auszeichnungsstriche, Paragraphzeichen und Unterstreichungen. Bl. 1r–51v am oberen Blattrand Angabe des jeweiligen Buches in Rot. Zweizeilige rote Lombarden, tw. mit Punktverdickungen. Bl. 1ra siebenzeilige rot-tintenfarbene kopfstempelförmig gespaltene Lombarde Q mit etwas ausgeschmücktem Caudaausläufer, Bl. 10ra und rb jeweils vierzeilige rot-tintenfarbene Initiale mit kopfstempelförmigen Aus sparungen. Bl. 52v–55v nur Lombarden und Überschriften in Rot. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, mit Goldbordüren gerahmt. Im zweiten Feld Golddruck M. S. DIAL. S. GREG. in Goldrahmung, im dritten Si gnaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 56r Besitzvermerk 15. Jh. Iste liber est monasterii s. Marie ad Novacellam prope Brixinam. Nach dem Inhaltsverzeichnis in Neustift, Stiftsbibl., Cod. 158, Bl. Ir gehört Cod. 83 zu einer Gruppe von urspr. tw. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Handschriften und Inkunabeln gemacht wurde) in sieben Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 H 15 (Bleistift) und Nr. 83/(1) (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 83/(1). 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 236, 283. – Stefani 96. 1 (Ir–v) leer. (1ra–52rb) Gregorius Magnus: Dialogi = Dialogorum libri IV de vita et miraculis patrum Italic orum (PL 77, 149–429 [Lib. 1, 3 und 4] bzw. PL 66, 126–204 [Lib. 2]. Ed. U. Moricca [Fonti per la storia d’Italia 57]. Roma 1924. Ed. A. de Vogüé [Sources chrétiennes 260]. Paris 1979 [Lib. 1–3] und [Sources chrétiennes 265]. Paris 1980 [Lib. 4]. – CPL Nr. 1713). 114 Cod. 85 (1ra) Lib. 1. – (10ra) Kapitelübersicht zu Lib. 2. – (10rb) Lib. 2 = Vita s. Benedicti abbatis. – (22vb) Kapitelübersicht zu Lib. 3. – (23rb) Lib. 3. – (38ra) Kapitelübersicht zu Lib. 4. – (38vb) Lib. 4. (52rb) Datierung: Millesimo quadringentesimo anno in vigilia sancti Ypoliti martiris (12. August 1400). 2 (52va–55va) ‹Ps.-Augustinus› (= ‹Ps.-Bernardus Claraevallensis› = ‹Ps.-Hieronymus› = ‹Ps.-Aegidius Romanus›): Speculum peccatoris (PL 40, 983–992. – Bloomfield Nr. 4918. R. Ru dolf, Ars moriendi. Von der Kunst des heilsamen Lebens und Sterbens [Forschungen zur Volkskunde 39]. Köln, Graz 1957, 14 Anm. 12. Weber I 174, Nr. 14; II 112. Zumkeller Nr. 65). Tit.: Speculum peccatoris. 3 (55va–b) ‹Bernardinus Senensis› (?): Speculum peccatorum sive amatorum mundi, unvollstän dig (Druck: GW 3889–3892. S. Bernardini Senensis Opera III. Venetiis 1745, 437–440, hier 437. – Bloomfield Nr. 6443. Th. Hohmann, Initienregister der Werke Heinrichs von Langenstein. Traditio 32 [1976] 399–426, hier Nr. 266). Tit.: Sermo per quondam Karthusiensem editus. (rot:) Sermo de conteptu (!) mundi et dicitur spe culum amatorum mundi. – Expl. mut.: … spaciosa via que ducit ad perdicionem ubi est perdicio nature et glorie [ Zur Verfasserfrage (auch Nicolaus de Dinkelsbühl, Iodocus de Heilprunna und Henricus de Langenstein zugeschrieben) s. Madre 297 (mit Angabe der Überlieferung). (56r) Besitzvermerk (s. bei G). (56v–59v) nur Rahmung. (Ir–v) leer. U. S. Cod. 85 TRACTATUS DE SACRAMENTIS Pap. I, 107, I Bl. 290 × 205. Wohl Neustift, Anfang 16. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + V10 + (VII+1)25 + (VI+1)38 + V48 + (VI+1)61 + VI73 + VII87 + VI99 + IV(107) + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. 11, 38 und 49 Einzelbl. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 215/225 × 120/125, tw. von feinen Tinten-, tw. von Blindlinien gerahmt, zu 35–42 Zeilen. Humanistische Kursive. A: Äußerst spärliche Rubrizierung: vereinzelte Auszeichnungsstriche; wenige zwei- bzw. dreizeilige Lombarden mit Punktverdickungen, tw. mit cadellenartigen Ausläufern. Repräsentanten. Bl. 1r P-Lombarde mit kopfstempelartigem Besatz und in gestrichelte Achterschlingen gelegtem, mit freien Kreisen sowie Fibrillen dekoriertem Ausläufer (vgl. Cod. 194, Bl. 22r und Cod. 405, Bl. 160v). E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe, Neustift, 2. H. 18. Jh. Pap. tw. abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck DE SACRAMENTIS in Goldrahmung, im dritten Signaturschild (s. bei G). Kapitale mit naturfarbenem und grü nem Zwirn umstochen. Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`4 D 11 (?, radiert, Tinte), II 1 H 17 (Bleistift) und N. 85 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 85. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedens Cod. 86 115 vertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol I 237, 283. (Ir–v) leer. (1r–106v) Tractatus de sacramentis. Inc.: Piissimus salvator Ihesus Christus tociusque consolator Deus omnis sue pietatis tantam humani generis imperitiam et ignaviam necnon infirmitatis … – Expl.: … et partim potest haberi ex dictis hic. (107r–v) nur Rahmung. (Ir–v) leer. P. A. Cod. 86 THOMAS DE AQUINO Perg. I, 40, I Bl. 300 × 205. Frankreich (?), wohl Mitte 13. Jh. B: Feines Perg., Haar- und Fleischseite deutlich unterscheidbar. Bl. 26 Loch mit Nahtspuren, Bl. 4 Nahtspur. Bl. 38 und 39 am oberen und unteren inneren Blattrand jeweils ein kleines Pergamentstück herausgeschnit ten (ohne Textverlust). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 2.VI24 + V33(34) + 3.I39(40) + (I–1)I* (Nach satzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Urspr. wohl durchwegs Sexternionen: Bei Lage 3 fehlt das äußerste Doppelbl.; nach Lage 3 fehlen wohl mehrere Lagen, von Lage 4–6 jeweils nur ein Doppelbl. erhalten (jeweils Textverlust). Bl. 35v Reklamante, beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 30 doppelt angeführt. S: Schriftraum 215 × 140. Zwei Spalten, von feinen Bleistift- bzw. Blindlinien gerahmt, zu einheitlich 50 Zeilen auf Bleistift- bzw. Blindlinierung. Am oberen und ab Bl. 3r zusätzlich am unteren Blattrand jeweils eine von Bleistift- bzw. Blindlinien gerahmte Zeile. Zirkelstiche. Textualis. A: Rote Auszeichnungsstriche; auf dem oberen Seitenrand der Rectos jeweils die Nummerierung der Text abschnitte in roten und blauen römischen Zahlen (auf dem Verso jeweils ein rotes DI für „Distinctio“); rote und blaue Paragraphzeichen; Verweiszeichen; rote Unterstreichungen. Zweizeilige rote und blaue Initialen mit Fleuronnée in der Gegenfarbe. Zu Beginn der Distinctiones fünf- bis achtzeilige Initialen (Bl. 1ra, 9ra, 11vb, 15va, 19ra, 20rb, 22rb, 24rb, 26va, 29rb, 32va) mit rot-blauem Fleuronnée von derselben Hand. Nur die Eingangsinitiale Bl. 1ra in den Schriftspiegel integriert, alle anderen Initialen (auch die kleineren) fast vollständig auf den Seitenrand bzw. ins Interkolumnium gerückt. Buchstabenkörper geometrisch, oft kopfstempelförmig rot-blau (Bl. 1ra ein farbig blau) gespalten. Fleuronnée meist in der Gegenfarbe gepunktet und gekernt; Kerne oft mit kleinen geschwungenen Fadenausläufern (z. B. Bl. 20rb). In den Binnenfeldern überwiegend Voluten, die selten (z. B. Bl. 1ra) Palmetten, Knospen und lilienartige Dreiblätter einschließen. Als Besatz oft Perlen-Dreier gruppen und Fäden, die sich häufig haarnadelförmig zurückbiegen; diese Fadenformationen (mit verbinden den Perlengruppen aneinandergereiht) bei kleineren Initialen auch als Ausläufer, die an den Enden mitunter (z. B. Bl. 35va) Fadensterne sowie kleinere dazwischengeschaltete Perlen und Doppelstriche als Dekor auf weisen. Bei größeren Initialen Fleuronnée-Leisten, die v. a. mit abwechselnd roten und blauen Kreissegmenten besetzt sind, welche in orthogonal abstehenden Fibrillen enden; als Endmotive überwiegend Perlengruppen; Bl. 32va Fleuronnée-Leiste zudem von einer Reihe kleiner Perlen begleitet (dieses Motiv auch an den Aus läufern kleinerer Initialen, z. B. Bl. 24rb). Das Fleuronnée könnte in Frankreich entstanden sein. Es zeigt einige Charakteristika der Pariser Produktion aus dem 2. Viertel 13. Jh. mit Tendenz zur Jahrhundertmitte (vgl. P. Stirnemann, Fils de la vierge. L’initiale à filigranes parisiennes 1140–1314. Revue de l’art 90 [1990] 58–73, hier 67–68, insbes. Cat. 25–31 mit Abb.). S. R. 116 Cod. 86 E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Insbes. am VD Pap. und Pappe tw. stark abgerieben und beschädigt. Am VD und HD Ecken mit braunem Leder verstärkt, tw. beschädigt. Rücken: braunes Leder, oben beschädigt, vier einfache Bünde, von Gold bordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten S. THOMÆ in Goldrahmung, im dritten PARS SUMÆ, ebenso in Goldrahmung, im vierten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`5 (?) C 7 (Tinte, radiert), II 1 H 18 (Bleistift) und N. 86 sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 86. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Shooner II 53. – Katalog ULB Tirol I 237, 283. – Steer 36. (Ir–v) leer. (1ra–39vb) Thomas de Aquino: Super librum secundum Sententiarum, unvollständig (Druck: u. a. GW M46376. Ed. R. P. Madonnet. Parisiis 1929. – Glorieux Nr. 14f2. Grabmann 286–290. Stegmüller RS Nr. 846, II. Stegmüller RS [1937] Nr. 1597. Shooner Nr. 1104). (1r) Tit.: Incipit super secundum fratris Thome. Dist. 1–6, 8–9 vollständig. Zahlreiche Fehlstellen: Dist. 7, 10, 22, 25, 41, 42 und 44 nur tw. erhalten. Dist. 11–21, 23–24, 26–40 und 43 fehlen zur Gänze. Reihenfolge tw. abweichend: Dist. 22 und 25 nachgereiht, Dist. 42 nach Dist. 44. (1ra) Prolog. Inc.: Spiritus eius ornavit celos et obstetricante manu eius eductus est coluber tortuosus Iob XXVI (13). Creaturarum consideratio pertinet ad theologos et philosophos … (1rb) Dist. 1. – (9ra) Dist. 2. – (11vb) Dist. 3. – (19ra) Dist. 4. – (20rb) Dist. 5. – (22rb) Dist. 6. (24rb) Dist. 7. (24vb) Expl. mut.: … nec mala voluntate carere possunt nec pena sed [ – (25ra) Inc. mut.: ] cognicio rei nisi per speciem eius sed species eventus futuri non potest … (26va) Dist. 8. – (29rb) Dist. 9. (32va) Dist. 10. (33vb) Expl. mut.: … non mediantibus principatibus qui hoc per se exequuntur [ (34ra) Dist. 41. Inc. mut.: ] enigma vel obscuritas sive imperfectio … – (34vb) Expl. mut.: … vocat irascibile et concupiscibile que sunt [ (35ra) Dist. 44. Inc. mut.: ] esse scilicet secundum quid aliquas prelationes a Deo non esse … – (35vb) Expl. mut.: … est medium inter timorem et audaciam scilicet considerat in omnibus [ (am unteren Blatt rand Reklamante: actibus) (36ra) Dist. 42. Inc. mut.: ] dicitur in Deum peccare … – (37vb) Expl. mut.: … maiori vel minori con temptu potest facere et huius ratio [ (38ra) Dist. 22. Inc. mut.: ] secundo an excuset peccatum. Ad primum sic proceditur … – (38vb) Expl. mut.: … que quis scire tenetur alias non quia etiam ha[bere (39ra) Dist. 25. Inc. mut.: ] malum quia li[berum] ar[bitrium] per se in bonum ordinatum est … – (39vb) Expl. mut.: … et per modum istum omnes actus humani sunt [ (Ir–v) leer. A. P. / U. S. Cod. 99 117 Cod. 99 CLEMENS V. IOHANNES ANDREAE. IOHANNES XXII. Perg. I, 33, I Bl. 400 × 270. Bologna, 1. H. 14. Jh. B: Urspr. helles, an den Rändern abgegriffenes Perg., Haar- und Fleischseite deutlich unterscheidbar. Lagen: 1I (Vorsatzbl., Pap. 18. Jh.) + IV8 + 2.V28 + (III–1)33 + 1I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zu Bl. 29 herausgeschnitten (Textverlust). Am Beginn und am Schluss fehlt jeweils wohl mindestens eine Lage (Text verlust). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Bl. 33 Rest eines neuzeitlichen gelben Papiersignakels der ULB Tirol. S: Schriftraum 255 × 155. Zwei Spalten, von kaum wahrnehmbaren Bleistiftlinien gerahmt (Text bis Bl. 31ra von Kommentar umgeben). Text mit wechselnder Zeilenzahl, Kommentar zu meist 72–80 Zeilen auf Bleistiftlinierung. Bl. 31v–32r Schriftraum (nur Text) 280 × 155. Zwei Spalten, von kaum wahrnehmbaren Bleistiftlinien gerahmt, zu 55 Zeilen. Bl. 32v–33v Schriftraum (nur Text) 320 × 200. Zwei Spalten, von kaum wahrnehmbaren Bleistiftlinien gerahmt, zu 82 Zeilen. Textualis formata (Littera Bononiensis), Text und Kommentar in unterschiedlicher Schriftgröße. Randglossen am seitlichen und unteren Blattrand (Gliederung mit Kleinbuchstaben) von verschiedenen Händen in Notula 15. Jh. A: Rote Überschriften, abwechselnd rote und blaue Paragraphzeichen (mit betont langgezogenem Oberstrich); Bl. 30va–32rb rote Auszeichnungsstriche, Paragraphzeichen und Zeilenfüllsel; in Tintenfarbe diverse Ver weiszeichen, Randmarkierungen und Zeigehände (von verschiedenen Händen). Ein- und dreizeilige rote und blaue Lombarden mit Fleuronnée in der Gegenfarbe. Repräsentanten. Fleuronnée: in den Binnenfeldern (und als Besatz) charakteristische saitenartige Parallelstriche, verziert mit kleinen Perlen und kleinbogig geränderten Blättchen; gelegentlich kurze Fadenausläufer; Bl. 33vb zu Be ginn der Extravagantes 16-zeilige rote I-Initiale mit bogenförmigen Aussparungen und blauem Fleuronnée; ebda die Explicit-Zeile der Summula mit wenig schwarzem Fleuronnée. Deckfarbenschmuck: in den Clementinen zu Beginn der Tituli vierzeilige figürliche Initialen, in der Glossa ordinaria vierzeilige ornamentale Initialen, jeweils mit wenigen Ausläuferblättern bzw. kurzen Ranken; am Anfang der Summula (Bl. 32va) zehnzeilige figürliche Initiale mit etwas mehr Rankenwerk. Blattvergoldete, schwarz konturierte Außengründe; Buchstabenkörper mit farblich abgesetzten Knoten motiven. Die Ranken(-Blätter) entspringen den Serifen oder setzen mittig und symmetrisch auseinander wachsend am Initialfeld an (Tropfenmotive in den Gabelungen), Adern und Schattierungen der Rankenblätter mit dunklerer Lokalfarbe deutlich herausgearbeitet, als Dekor schwarz konturierte Goldpunkte (mit feinen tintenfarbenen Fadenfortsätzen). Lebhafte Farbigkeit: neben Rosa, Rot, mehreren Grün- und Blautönen, Cremeweiß in Kombination mit senffarbigem Ocker und Grauviolett bis hin zu kräftigerem Violett; häufig feiner weißer Liniendekor. In den Binnenfeldern der Ornamentalinitialen zumeist Blattmotive (u. a. an Rankengabelungen erinnernd, Bl. 17rb); die figürlichen Initialen zeigen männliche Büsten (Ausnahmen: Bl. 12va, zum Tit. De baptismo et eius effectu, Rundgesicht, das wohl in Stein gemeißelt sein soll; Bl. 16ra, zum Tit. De magistris, statt I-Initiale ca. achtzeilige Halbfigur eines Bischofs, dessen Körper nach unten in Rankenblätter ausläuft), tw. als Kleriker oder Gelehrte charakterisiert (auch mit Textbezug, z. B. Bl. 5rb: Mönch, De statu monachorum et canonicorum regularium, Lib. 3, tit. 10, cap. 1); Gesichter mit horizontalen Augenbrauen und kleinen, dicklippigen Mündern, Inkarnat mit Grau abschattiert. Bl. 32va Unterrichtsszene (mit Autorbildnis): Der Lehrende (wohl Iohannes Andreae) in blauem Gewand mit rotem, pelzbesetztem Mantel (und ebensolcher Kappe) etwas erhöht auf einem Holzstuhl mit Querbrett, auf dem ein geöffnetes Buch und Schreibutensilien liegen; mit den dünnen Fingern der Rechten hält er ein Schreibgerät und wendet sich im Profil dem Auditorium (Mönchen?) zu. Der Deckfarbenschmuck entstand im 3. Jz. 14. Jh. in Bologna (freundliche Auskunft Ulrike Bauer-Eber hardt, München). In der Gestaltung der Hände (stäbchenartige Finger) sowie der Mund‑ und Augenpartien besteht eine gewisse Ähnlichkeit mit denjenigen Figuren im Volumen parvum Bordeaux, Bibliothèque mu nicipale, ms. 355.1, die einem Buchmaler aus dem Kontext des „Maestro del 1328“ zugeschrieben werden (G. del Monaco, I manoscritti miniati dell’Illustratore nella cultura figurativa del secondo quarto del XIV secolo, Diss. Bologna 2012, 159–162 [Nr. 3], http://amsdottorato.unibo.it/5083/: In dieser ca. 1335 datierten Hs. begegnen neben der bewussten Hand aus dem Umfeld des „Maestro del 1328“ auch der „Illustratore“ und der „Maestro dell’Authenticum di Bordeaux“). – Das Fleuronnée ist oberitalienisch. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. abgerieben und beschädigt. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck CLEMENTIN. 118 Cod. 99 (stark zerstört) in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Die Hs. zählt wohl zu jenen vier Hs. kirchenrechtlichen Inhalts, die 1353 als Geschenk des Herrn Berthold, Hospitaler zu Brixen, in den Besitz des Stiftes gelangten (vgl. Neustift, Stiftsarchiv, WW 59. Mairhofer 264–265; Peintner, Stiftsbibliothek 118; Peintner, Schreibkunst 377–378). Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`2 […] 6 (?, Tinte, radiert), II 2 C 3 (Bleistift); Nr. 99 (Tinte) und Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 99. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wretschko–Sprung 15. – Hermann 155, Nr. 110. – B. Pagnin, La „littera Bononiensis“, studio paleografico. Atti del Reale Istituto Veneto di scienze, lettere ed arti 93/2 (1933/34) 1593–1665, Nr. 24. – Furtenbach– Kalb 20. – Neuhauser, Neustift 88. – Katalog ULB Tirol I 279, 283. – J. Tarrant, The manuscripts of the Constitutiones Clementinae I: Admont to Munich. ZRG Kanonistische Abt. 70 (1984) 109 und II: Napoli to Zwettl. ZRG Kanonistische Abt. 71 (1985) 80. – Peintner, Stiftsbibliothek 118–119. – Peintner, Schreibkunst 377–378. 1 (Ir–v) leer. (1ra–32rb) ‹Clemens V.›: Constitutiones (Clementinen), unvollständig: enthält nur Lib. 3, tit. 8, cap. 2 bis Schluss (CIC, ed. Friedberg II 1162–1200. – Tarrant II 80). Mit der Glossa ordinaria des Iohannes Andreae (Druck: u. a. GW 7101). (1ra–32rb) Text. Inc. mut.: ] quasi torrens pregravemur negociis curis excitemur innumeris co gitacionibus … (CIC, ed. Friedberg II 1162, Z. 12) – Expl.: … Si quis autem hec actemptare (!) presumpserit indignacione omnipotentis Dei et beatorum Petri et Pauli apostolorum eius se noverit incursurum. (1ra–31ra) Kommentar. Inc. mut.: ]usurarum excommunicatum publice nominatum interdictum et tempus interdicti numquid una vel quadruplici excommunicacione ligabitur … (Druck: GW 7101, Sign. E3 Anm. n) – Expl.: … cum ad illam redibimus per intercessionem virginis gloriose nos collocet cum electis Amen. 2 (32va–33vb) Iohannes Andreae: Summula quarti libri Decretalium (Summa de sponsalibus et matrimonio) = Überarbeitung von ‹Iohannes de Anguissola›: Summa de sponsalibus et matri moniis (Druck: GW 1754. – Tarrant II 80). Inc.: Cristi nomen invocans ad honorem ipsius et reverentissimi patris domini Bononiensis archi diaconi qui divinam potenciam imitatus de me nichil fecit aliquid … – Expl.: … et non ad propo situm unde quid dicunt vide per te. Iohannes Andree. Explicit Summula quarti libri Decretalium Iohannis Andree. 3 (33vb) Iohannes XXII.: Extravagantes, unvollständig: Tit. 14, cap. 3 (Avignon, 1322 Dezember 8) (Druck: u. a. Lyon 1559, 114–118. CIC, ed. Friedberg II 1225–1226, Z. 6. J. Tarrant, Extra vagantes Iohannis XXII. [Monumenta iuris canonici B 6]. Città del Vaticano 1983, Nr. 18, 228–234). Beginn gegenüber Ed. und Druck leicht abweichend, jedoch in Übereinstimmung mit Variante RVC, s. Tarrant 134 bzw. 228, Apparat. Cod. 99 bei Tarrant nicht genannt. Inc.: Iohannes episcopus servus servorum Dei. Ad perpetuam rei memoriam valitura canonum. Non est dubium pertinere cum statuta a se vel predecessoribus suis edita obesse percipit pocius quam prodesse ne ulterius adesse valeant providere. Dudum si quidem felicis recordacionis Nicolaus papa IIIus predecessor noster attente prospiciens … – Expl. mut.: … quam si res illas cum illo quo carere se dicunt domino optinent [ Lit.: Schulte GQ II 133 und 214. (Ir–v) leer. W. N. Cod. 100 119 Cod. 100 MISSALE (FESTIVUM) Perg. 104 Bl. 380 × 295. Neustift, 1524–1526. B: Hs. 2017 restauriert: gereinigt, loses Bl. 99 wieder eingebunden, lockere Lagen stabilisiert (vgl. Restaurier protokoll). Haar- und Fleischseite deutlich unterscheidbar, bisweilen etwas gewellt, einzelne Löcher (Bl. 22, 40) und Nahtspuren (Bl. 37). Bl. 73rb Initiale herausgeschnitten (s. bei A). Lagen: 6.IV48 + I50 + 6.IV98 + (IV–2)104. Gegenbl. zu Bl. 99 und 100 herausgeschnitten (ohne Textverlust). Bei den meisten figürlichen Initialen Klebespuren und tw. Stoffreste von einstigen Schutztüchlein. Reste von dunklen Ledersignakeln. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 270/275 × 195/200. Zwei Spalten, seitlich von Tintenlinien begrenzt, zu einheitlich 28 Zeilen. Bl. 52r–61r (Canon missae) Schriftraum 265/275 × 195/200, von Tintenlinien gerahmt, zu einheitlich 15 Zeilen auf feiner Tintenlinierung. Zirkelstiche. Textualis formata (Textura), dat. 1524 (Bl. 38rb, 50rb, 50v, 93rb, 94va, 104ra). Schreiber: Stephan Stetner (Bl. 38rb, 93rb, 94va, 104ra), vgl. Krämer, Scriptores. N: Bl. 38v–47v sowie Bl. 56v–57r Schriftraum 260/265 × 200. Seitlich von Tintenlinien gerahmt, links jeweils mit eigener Spalte für den Notenschlüssel. Jeweils neun Notensysteme sowie Textzeilen. Gotische Choral notation mit f-Schlüssel auf vier roten Linien. Kustoden und rote Gliederungsstriche. A: Rote Auszeichnungsstriche, Überschriften, Durchstreichungen und Benediktionskreuze (rote Tatzen- und Kolbenkreuze, z. B. Bl. 56r). Bei den Datierungen und/oder Stetners Namensnennungen bzw. Initialen (s. bei S) jeweils fünf tintenfarbene Ziermotive bzw. Trennzeichen aus drei kleinen Kreisen in Dreiecksan ordnung mit kurzem, kommaartigem Fadenausläufer (Bl. 104ra zusätzlich Kreuz, vielleicht von späterer Hand als Sterbekreuz für den am 12. Dezember 1526 verstorbenen Stetner?); in den Binnenfeldern der Majuskeln gelegentlich feine Doppelschrägstriche, tw. mit einseitigem Bogenbesatz. Rote und blaue ein- bis vierzeilige Lombarden mit Punktverdickungen und kürzeren Ausläufern; einige aufwändigere (I-)Lombar den,deren Ausläufer in Achterschlaufen enden (z. B. Bl. 5va, 6va; sehr ähnliche Lombarden in Sign. 14820, vgl. z. B. Cod. 100, Bl. 5va mit Sign. 14820, Bl. 82v6, aber wohl nicht von einer Hand); v. a. die größeren Lombarden mit Fleuronnée: rote Lombarden mit hellblauem oder grünem, blaue Lombarden z umeist mit Fleuronnée in Rottönen, seltener in Grün und Lila. Repräsentanten. Fleuronnée: in den Binnenfeldern charakteristische zierliche Spiralranken (oft mit Punktenden), häufiger auch Blättchenmotive, Rautengitter und dachziegelartig angeordnete Bögen. Konturlinien. Wenige kurze Fadenausläufer, dafür die längeren Lombardenausläufer konturiert und mit Fleuronnée (v. a. Fibrillen) ver ziert; in den Ausläuferschlingen zuweilen Blattmedaillons; charakteristisch auch der Strahlenbesatz an kleinen Kreisen und Punktverdickungen (sowohl ins Binnenfeld als auch nach außen gerichtet); als Besatz außerdem Fibrillen, auch in Kombination mit Spiralen; derartige Fibrillen sowie kleine strahlenbesetzte Kreise auch in den Deckfarben-Randleisten. – Lombarden mit figürlichem Federzeichnungsdekor (Aus wahl): Maskarons (Bl. 35rb1, 97va) und maskaronartige Köpfe (Bl. 17rb1, 101vb); heraldische Vögel (Bl. 21ra Adler mit gespreizten Flügeln, 26rb doppelköpfiger Adler, 37rb3 aufflatternder Greif?); diverse Köpfe und Büsten, u. a. Bl. 46v Haupt Christi im Strahlenkranz, 40r Engelsbüste, 25vb zwei Greisenköpfe im Profil einander zugewandt, der linke mit geöffnetem Mund; Bl. 48va zwei Fangen spielende Putti. Mas karons und Haupt Christi sehr fein ausgeführt. Deckfarbenschmuck: sechs ornamentale und 23 figürliche Initialen, jeweils mit Randschmuck, ein Kuss bildchen sowie zwei Vollbilder mit Randdekor; insgesamt sieben Wappen mit kartuschenartigen Schilden. Die Ornamentalinitialen vorrangig im Winterteil des Temporale, im Ordinarium missae und in den Prae fationes (Bl. 2ra, 38v, 39r, 40v, 41v, 42r), siebenzeilig bzw. zwei Notenzeilen hoch, Randschmuck v. a. Typ 2 und 3 (s. u.). Auch die grob ausgeschnittene Initiale Bl. 73rb (Mariä Empfängnis) könnte eine ornamentale Initiale gewesen sein (nur fünfzeilig, kurze Ranken, insbes. Typ 2; muss vor 1905 ausgeschnitten worden sein, da Verlust bei Hermann 162 vermerkt). Die figürlichen Initialen im Temporale und Sanctorale (Bl. 3ra, 5rb, 7ra, 8rb, 9vb, 11va, 17rb, 43v, 45v, 52r, 62ra, 63va, 68va, 70rb, 71va, 76ra, 77rb, 79rb, 80vb, 83rb, 85ra, 86va, 88rb) in der Regel sechs- und siebenzeilig. Rahmen der Ornamentalinitialen einfarbig oder aus grünen, blauen, rosafarbenen oder roten Abschnitten zusammengesetzt; dieser Rahmentyp selten bei figürlichen Initialen, die zumeist von breiteren, blattvergol deten Rahmen umgeben sind (Konturierung und Profilierung mit schwarzen und weißen Linien); nur die Or namentalinitiale Bl. 42r mit (einfach punziertem) goldenem Rahmen. Breitere unverzierte Goldrahmen auch bei den Vollbildern. – Buchstabenkörper häufig aus Renaissancemotiven wie Kandelabern, Säulen, Frucht schalen und -körben, Festons, Rüstungsteilen (Brustpanzern), Blattmasken, Fabelwesen (v. a. Drachen) und Puttenköpfen (geflügelt) aufgebaut, bei den Ornamentalinitialen gerne in Lila- und Violetttönen koloriert, 120 Cod. 100 bei den figürlichen Initialen oft in Gold mit schwarzen Kontur- und Binnenlinien, Schraffuren auch in Weiß; darüber hinaus glatte goldene Buchstabenkörper mit minutiösem, in weißlichem Malmittel ausgeführtem Dekor, s. Te-igitur-Initiale Bl. 52r (auf dem Schaft z. B. zartes Flügelpaar); nur wenige Buchstabenkörper, die in spätmittelalterlicher Manier mit ondulierenden Blattranken gefüllt sind. – Initialfelder der Ornamen talinitialen blattvergoldet (bis auf Bl. 42r: blaugrundig) und mit Rautengittern aus Punktlinien (gefüllt mit Blütenpunzen) überzogen; bei den figürlichen Initialen Bl. 7ra, 8rb, 9vb, 17rb, 52r, 62ra, 68va, 70rb, 76ra und 80vb jeweils Miniatur im Initialfeld, Buchstabenkörper davorgesetzt (nur Bl. 70rb Initialbuchstabe auf den Rahmen appliziert); wenn das Bild im Binnenfeld platziert und die Initiale von einem farbigen Außen grund (meist kobaltblau oder dunkelrot; Bl. 83rb Wolkenhimmel) umgeben ist, dann können Rahmen und Buchstabenkörper Schatten auf den Farbgrund werfen. Variationsreicher und üppiger Randschmuck, der nur eine Ecke des Schriftspiegels umschließt, aber auch zwei bis vier Seitenränder bedecken kann; vier Haupttypen: 1. kandelaberartige Randleisten, die meist auf Sockeln, Säulen, vasenartigen Gefäßen etc. ruhen und sich aus Renaissancemotiven (s. o. bei Buchstaben körpern, zusätzlich bukranienartige Blattmasken, Waffen, heraldisch anmutende Flügelpaare, Musikinstru mente, Schmuckstücke; zu figürlichen Komponenten s. u. Detailbeschreibungen) im Wechsel mit floralen Elementen (Blüten und breite Blätter mit volutenartig eingerollten Spitzen) symmetrisch aufbauen; die ein zelnen Abschnitte der Achsenstäbe leicht kolbenförmig. V. a. am Beginn der Hs. (bis Bl. 8r), beim Kanon bild und am Anfang des Messkanons, meist auf beiden Vertikalrändern des Folios; die äußeren Kandelaber leisten, die gesamte Marginalfläche nutzend, breiter angelegt und häufig mit einem Fruchtkorb bekrönt; 2. dünnastige, gerne symmetrisch angelegte Spiralranken mit bunten, fein gezackten Blättern, Fantasie blüten, Goldscheibchen und charakteristischen Perlreihen u. a. als Fruchtstand; z. B. Bl. 2r, rechter Seiten rand; 3. farbenfreudige Akanthusranken, oft gerade geführt, mit Eck- bzw. Endschlingen, die mit (punzier tem) Blattgold gefüllt sind; wie bei den Kandelaberleisten Tendenz zu symmetrischem Aufbau; als Ziermotiv goldene Manschetten; nur dieser Rankentyp setzt, oft aus floralen Manschetten entspringend, direkt und mittig am Initialrahmen an (lediglich bei der Oster-Initiale Bl. 62ra spätmittelalterliche Gestaltungsweise: Ranken entwachsen den Serifen des Buchstabenkörpers, der hier zudem mit Blattwerk gefüllt ist, s. o.); als Beispiel s. Bl. 11v; 4. naturalistische (Blüten-)Zweige, die botanisch identifizierbar sind (s. u. Detail beschreibungen); verziert mit Fibrillen und (punzierten) Goldfeldern; Kombination von naturalistischen Zweigen und stilisierter Goldzier als Charakteristikum anzusehen; diese Zweige häufiger bei figürlichen als bei ornamentalen Initialen. – Die Typen 2, 3 und 4, die auch als Mischformen auftreten können, zusätzlich mit Drolerien belebt (s. u. Detailbeschreibungen). Zudem detailfreudige Bas-de-page-Illuminationen (s. u. Detailbeschreibungen), bei denen wie bei den Drolerien eine Vorliebe für Jagdszenen festzustellen ist. Auf die Tätigkeit einer Meisterhand (und wenigstens einer Assistenzhand) dürften u. a. folgende Figuren mit bes. differenziert gemalten Gesichtszügen, Haaren und Gewändern hinweisen (ausführlich zur Händeschei dung unter sorgfältiger Berücksichtigung stilistisch-technischer Charakteristika s. Fiegl, Missale 118–120, die durchaus drei Miniatorenhände für möglich hält): Bl. 1v die drei Hauptfiguren, dabei Augustin Posch mit Porträtcharakter und das Mariengewand mit bes. nuancierter Draperie; Bl. 51v alle drei Figuren der Kreuzigungsgruppe; Bl. 52ra Abraham; Bl. 58v Antlitz Christi auf dem Schweißtuch (man beachte zudem die Drapierung des Tuches und die Gestaltung der Engelsgewänder) und Bl. 70rb Gottvater. Auch die Basde-page-Szenen Bl. 52r und 79r, bei denen die Auswirkungen des Windes nicht nur durch wehende Haare spürbar gemacht werden, sowie die Finesse der Epiphanie-Initiale Bl. 8r sind bemerkenswert. Generelle Charakteristika: Figuren in allen Körperhaltungen anatomisch sehr gekonnt dargestellt, nur Kniepositionen etwas unsicher (z. B. Augustin Posch, Bl. 1v). Heiligenscheine häufig als goldene, schwarz konturierte Scheiben, die bei Figuren in Rückenansicht tellerartig am Hinterkopf sitzen (z. B. Bl. 63va); Engel und Putti mit buntfarbigen Flügeln. Gebauschte, reich gefältelte und fallweise abflatternde Gewänder. Sehr sichere (Zentral-)Perspektive, nur wenige Ungereimtheiten (z. B. Bl. 7ra Thronlehne nicht in Fluchtlinie, Altar mensa zu aufsichtig; Bl. 68va Kapitelle der rechten Säulenreihe verschoben); Vorliebe für tonnengewölbte Räume; gekonnter Einsatz von Hell und Dunkel bis hin zu subtilen Schattenwürfen (z. B. Bl. 52r Schatten des Feuerholzes auf der Altarfront und Bl. 86v Schatten des Hirschkäfers auf dem Bas-de-page) und trompel’œil-artigen Schattenwürfen an Initialen (s. o.). Kleinteilige Darstellung von Mauerwerk (z. B. unregelmä ßige Struktur der Ruinenmauern Bl. 8rb) und Marmor (Altäre). Naturgetreue Gestaltung von Böden mit Steinen, Grasbüscheln, Wegen etc. Als Hintergründe in den Miniaturen gerne Landschaftsausblicke mit Gebirgspanoramen, Burgen oder Stadtveduten (s. v. a. Kanonblatt Bl. 51v), Himmel mit nuancierten Wolken strukturen (z. B. Bl. 52ra, hier auch die aus dem Feuer aufsteigenden Rauchschwaden sehr gekonnt). Strahl endes Kolorit (s. auch Hermann 156) u. a. Kobaltblau, Violett mit Tendenz zu Pink und Orangegelb (dieses nicht zuletzt für Judasgewand im Letzten Abendmahl, Bl. 17rb); die Assistenzhand (bei Figuren- und Land schaftsdarstellungen) mit stärkerem Farbauftrag als die Meisterhand (Fiegl, Missale 119); reiche Verwen dung von Blattgold auf beigefarbenem Bolus (sichtbar z. B. Bl. 52r T-Initiale). Einfache Punzierungen: Punktlinien und kleine Kreise, die zu Rosetten zusammengesetzt sein können. Figürliche Initialen und erwähnenswerter Randschmuck (auch auf Folios mit ornamentalen Initialen): Cod. 100 121 Bl. 2r [Ornamentalinitiale] – Auf dem oberen Seitenrand Neustifter Wappen: in Rot goldenes Tau-Kreuz. Bas-de-page: mittig ein Kandelaber, Ranken (Typ 2, mit Puttenköpfchen), in deren Schutz vier Putti, beglei tet von einem springenden Hund, derbe Späße treiben. Drolerien: Eichhörnchen, drei Vögel. Bl. 3ra Geburt Christi: Maria und Josef beten kniend das auf dem Boden (auf einem roten Tuch) liegende nackte Kind an, das die Händchen nach Maria ausstreckt; neben Josef, der eine Kerze hält, Ochs und Esel; am Himmel vier Putti, die von einem Notenblatt singen; im Hintergrund auf einem Mauerrest ein rötlicher Säulenstumpf. – Bas-de-page: Ranken (Typ 2), davor Putto auf der Hirschjagd, der im Laufen ein Jagdhorn bläst und von einem angeleinten und zwei frei laufenden Hunden, die den Hirsch attackieren, unterstützt wird. In der rechten Kanderlaberleiste zwei Posaunenengel. Drolerien: Kröte, drei Vögel. Bl. 5rb Jesuskind in hauchfeiner Strahlenmandorla auf hellblauer Wolke stehend (linker Fuß leicht angeho ben), in der Linken die Weltkugel, Zeigefinger der Rechten erhoben. – Bas-de-page: mittig Putto, der auf einem Sockel stehend einen Kandelaber balanciert, Ranken (Typ 2), unter deren Bögen zwei Hirsche liegen. In der rechten Kandelaberleiste Wappen: vor zwei gekreuzten Säbeln blauer Schild (von goldenem Rauten gitter überzogen), darauf goldener Zweig mit Blättern (Hermann 158: Erdbeerblatt); unter der bekrönenden Fruchtschale Gruppe von sieben Putti, die von einem Notenblatt singen. Bl. 7ra Beschneidung Christi: In einem sakralen Raum (im Hintergrund die Gesetzestafeln auf einem Altar) auf einem thronartigen Sitz ein greiser Priester, der das Jesuskind auf dem Schoß hält, das vom Mochel be schnitten wird; daneben Assistenzgruppe, angeführt von Maria und Josef; Boden u. a. mit Adlerfliesen belegt. – Bas-de-page: Drei Putti mit Jagdhörnern, Speer bzw. Stange und Fangnetz jagen, begleitet von vier frei laufenden Hunden, ein Hasenpaar, dem ein Igel voranläuft, auf dessen Stacheln Früchte stecken. Die Kandelaberleiste links ruht auf männlicher Büste mit Tatzen; im oberen Bereich der Leiste Neustifter Wappen: in Rot goldenes Tau-Kreuz. Rechte Randleiste: oben Blütenranke, unten Weinstock: Reben mit blauen Trauben ranken sich um rote Stange, an der eine kleine Sichel klemmt. Drolerien (Weinstock): zwei blau-gelbliche Vögel, zwei Biber. Bl. 8rb Anbetung der Heiligen Drei Könige: In einer Ruinen-Szenerie sitzt Maria mit dem nackten Kind auf dem Schoß, das nach der Kiste mit Gold greift, welche ihm der greise, vor ihm kniende König darbringt; dahinter die beiden anderen Könige mit ihren Gaben und Gefolge; am Himmel der Stern der Weisen. – Bas-de-page: Soldat mit weiß-blauer Fahne; Ranken (Typ 2) mit vier Drolerievögeln, u. a. Wiedehopf und grüner Papagei. In der linken, von einem geflügelten Puttenkopf bekrönten Randleiste kleines Bildnis der sich erstechenden Lucretia. Die rechte Randleiste ruht auf kniendem Schildträger (mit Brustpanzer), darüber vor gekreuzten Waffen ein Rundschild mit männlichem Profilkopf, des Weiteren ein kniender Putto mit zwei Füllhörnern und an der Spitze ein Bildchen des dornengekrönten Christus (auf blauem Grund, in Architek turrahmen). Bl. 9vb Darbringung Christi im Tempel: Vor einem mit einem feinen Tuch bedeckten Altar, auf dem sich die Gesetzestafeln erheben, reicht Maria das Kind dem Priester (mit Goldmitra); neben Maria stehen Josef und eine Frau mit zwei Opfertauben, hinter dem Priester drei jüdische Greise. – Unterhalb der Initiale Rankentyp 3 (und 2), daneben naturalistischer Zweig mit gelben Blüten, auf dem linken Seitenrand Blütenranke (vgl. Bl. 7r) und darüber weiterer naturalistischer Zweig mit länglichen, roten Blüten. Drolerien: Vogel und roter Käfer. Bl. 11va Einzug in Jerusalem: Jesus reitet segnend auf der Eselin dem Stadttor zu, vor dem Männer mit Palmzweigen stehen; hinter ihm die Apostel, vor ihm breitet ein Mann in kostbarem Gewand einen roten Mantel aus; das Gesicht Jesu abgerieben. – In den Ranken (Typ 3) ein Marder, der einen rosafarbenen Vogel anfaucht, der sich nach seinem Jäger umwendet. Bl. 17rb Letztes Abendmahl: Jesus und seine Jünger an einem mit feinem, weißem Tuch gedeckten Tisch; Johannes kauert auf dem Schoß Jesu, der dem gegenüber sitzenden Judas (mit rotem Beutel in der Linken) einen Bissen reicht; im Vordergrund schenkt ein Jünger Wein aus einer Zinnkanne in einen Becher, neben ihm auf dem Fliesenboden Kupferbecken mit einer weiteren Kanne; im Hintergrund großes Fenster mit Ar chitekturrahmung. – Als Randzier rote Disteln und Weinranke, die in Rankentyp 3 endet. Drolerien: fasanen artiger, blau-gelb-roter Vogel; blaue Libelle. Bl. 38v [Ornamentalinitiale] – Rankentyp 3 mit Dompfaff. Bl. 39r [Ornamentalinitiale] – Rankentyp 3 mit zwei Vögeln sowie ein Zweig kleiner, roter Nelken, darin eine Henne mit ihren Küken. Bl. 40v [Ornamentalinitiale] – Rankentyp 3 mit Uhu (auf einem Bein stehend, mit gespreizten Flügeln), der von drei Vögeln attackiert wird. Bl. 41v [Ornamentalinitiale] – Rankentyp 3 mit Maikäfer, bräunlichem Falter, Fliege und zwei Vögeln. Bl. 42r [Ornamentalinitiale] – Rankentyp 3, auf dem Bas-de-page in Blütenzweig (vgl. Bl. 7r) auslaufend; graubrauner Falter. Bl. 43v Sitzendes Äffchen an einer Leine, das sich in Rundspiegel betrachtet. – Rankentyp 3 mit Libelle und Elster. 122 Cod. 100 Bl. 45v Halbfigur des segnenden Christus mit Weltkugel in der Linken, auf blassblauer Wolke, umgeben von gelb-pinkfarbener Aureole. – Als Randzier Wiesensalbei- und Butterblumenzweig, ineinander gehakt, mit grau-gelbem Vogel. Bl. 52r Opferung Isaaks: Isaak kniet betend auf einem Altar, neben ihm auf der Mensa brennt rauchend ein Feuer. Abraham berührt mit der Linken die Schulter seines Sohnes, wobei sein ausgestreckter Arm vom TSchaft fast verdeckt wird, und erhebt mit der Rechten das Schwert, woran ihn ein herbeifliegender Engel hindert; zw. Engel und Abraham der im Gesträuch verfangene Widder. – Auf dem Bas-de-page zugehörige Szene: In einer liebevoll gestalteten Seeuferlandschaft schreiten Abraham (mit Fackeln) und Isaak (mit Holzbündel), begleitet von einem Hund, gegen den Wind an, der ihre Kleider und Haare wehen lässt, Bäum chen umbiegt und Vögel über den Himmel jagt. Mittig auf dem oberen Seitenrand detailliert ausgeführte Blattmaske mit Ranken (Typ 2), die große Ähnlichkeit zu dem mit der Feder gezeichneten Maskaron Bl. 97va aufweist. Die linke Kandelaberleiste wird von einem knienden, grauhaarigen Mann mit Rüstung getragen, am Fuß der rechten Kanderlaberleiste zwei musizierende Satyrn (abgegriffen, s. bei B). Bl. 62ra Auferstehung Christi: Der Auferstandene (mit Wundmalen) steht im Kontrapost mit wallendem, rotem Mantel, Siegesfahne und Strahlennimbus segnend auf dem geschlossenen Sarkophag. Davor stützt sich einer der schlafenden Soldaten auf einen großen, hölzernen Humpen. Die Strahlen der im Hintergrund über der Hochgebirgskette aufgehenden Sonne korrespondieren mit dem Heiligenschein Christi. – Zentral auf dem Bas-de-page Pfau, der ein Rad schlägt, links neben ihm Pfauhenne und herbeifliegender Fasan (vor Ästchen mit zart lilafarbenen Blüten); rechts neben dem Pfau Zweige mit Feigen, unter denen ein Putto schläft und zwei Affen herabgefallene Früchte aufsammeln und fressen; davor eine Maus. Auf dem rechten Seitenrand Ranke aus folgenden Abschnitten: unten Eichenzweig, von dem ein Putto mit umgehängter Tasche Eicheln pflückt; in diesen Zweig eingehakt stachelige Heckenrosenranke, in der ein Laute spielender Putto sitzt; an diese Ranke eine Blütenranke (vgl. Bl. 7r) mit Hilfe eines roten Bändchens angebunden, zuoberst Rabennest, aus dem ein Putto eines der Rabenküken gestohlen hat und einem weiteren Engelchen, das ihm mit einer Schale oder einem Hut entgegen klettert, reichen will, was die heranfliegende Rabenmutter zu verhindern versucht. Darüber, auf dem oberen Seitenrand, verfolgt ein Greifvogel eine Wildente. Von der Initiale ausgehend Ranken (Typ 3); darunter, auf dem linken Seitenrand, frei stehendes Maiglöckchen; zuunterst naturalistisches Zweiglein mit roten Blüten. Bl. 63va Himmelfahrt Christi: Maria und Apostel knien um Auffahrtsberg (darauf Fußspuren Christi) und blicken zum Himmel empor, wo Beine und Gewandsaum des Auffahrenden im Strahlenkranz zu sehen sind. Beine Christi abgerieben. – Als Randzier Akelei- und Heckenrosenzweig (mit kleinen Dornen), ineinander gehakt. Drolerien: Käfer, roter Falter, stäbchenförmiges rotes Insekt und ein Vogel. Bl. 68va Pfingsten: Maria und Apostel in einer Säulenhalle, über ihnen die Taube des hl. Geistes (von Strah lenkranz umgeben) in wolkenumsäumter, gelb-rosafarbener Aureole. – Auf dem Bas-de-page großer, von einem Putto (mit Schwert) geführter Braunbär, der den Betrachter mit gefletschten Zähnen anblickt. Auf ihm reitet ein Posaune blasender Putto mit purpurfarbenem Kopfband, das entgegengesetzt zu den an der Posau ne befestigten Bändern abflattert. Musikalisch begleitet wird der Zug von zwei Putti, einer davon mit hoher Kappe, die Krummhorn und ein Streichinstrument spielen; über ihnen, in einem Bäumchen, eine Elster, am Himmel größerer schwarzer Vogel, der einem hellblauen Vögelchen nachstellt. Am Bäumchen angebunden ein Affe, hinter dem ein Feigenast emporwächst; im Vordergrund Frosch und Schildkröte. Auf dem oberen Seitenrand jagt ein Greifvogel einen Stieglitz. Um die linke obere Seitenecke legt sich ein Zweig mit kleinen blau-roten Blüten, in den eine große Iris kopfüber eingehängt ist. Bl. 70rb Gnadenstuhl: Gottvater sitzt, mit der Taube des hl. Geistes auf der Schulter, auf einem Marmorthron und hält, seinen Blick zur Seite gerichtet, den Leichnam seines Sohnes vor sich, der den Betrachter unter halb gesenkten Lidern anzublicken scheint. – Bas-de-page: Rankentyp 2. Bl. 71va Zwei Engel präsentieren eine große Monstranz mit Hostie. – Als Randzier naturalistischer Zweig mit roten Blüten (Mohn?) und Rankentyp 3, der in Typ 2 übergeht. Drolerien: Vogel, kleine Libelle, Fliege (?) und Maikäfer. Bl. 73rb [Ornamentalinitiale?] – Rankentyp 2 (und 3). Drolerien: grüner Papagei und Greifvogel, der sich auf eine Wildente herabstürzt. Bl. 76ra Verkündigung: In einem schlichten und trotz zweier Fenster relativ düsteren Raum kniet Maria mit vor der Brust gekreuzten Armen an einem hölzernen Betpult (mit Fronttürchen), auf dem ein aufgeschlage nes Buch liegt; über ihr die Taube des hl. Geistes im Strahlenkranz, neben dem Pult bauchige Bodenvase mit Maiglöckchenstrauß. Von links eilt segnend der Engel herbei, in der Linken einen Stab, um den sich das Spruchband Ave Ma[ria] gr[a]c[i]a plen[a], d[ominus] te[cum] windet. – Linker Seitenrand und Bas-de- page: Rankentyp 3 mit Marienkäfer, Heuschrecke und blau-rotem Vogel. Rechter Seitenrand: unten Blüten ranke (vgl. Bl. 7r), darin eingehängt oben Ranke Typ 3 (und 2). Bl. 77rb Heimsuchung: Vor einem rundbogigen Tor (darüber Fachwerk-Erker) begrüßt Elisabeth Maria. – Rankentyp 3 mit Vogel, Insekt und Fuchs, der eine Blaumeise bedroht und dabei von einer Elster beschimpft wird. Cod. 100 123 Bl. 79rb Marientod: Die verstorbene Maria sitzend, mit hoher, brennender Kerze in der Rechten in einem Himmelbett; davor drei betende bzw. lesende Apostel; ein Apostel hebt den Bettvorhang an und berührt Maria am Ellbogen, Johannes unterstützt Maria beim Halten der Totenkerze, hinter ihm Petrus mit Aspergil und geöffnetem Gebetbuch, gefolgt von den anderen Aposteln, von denen der vorderste den Weihwasserkessel trägt; über Johannes das Seelenfigürchen Mariens, das in den Himmel zu Christus emporschwebt. – Auf dem Bas-de-page zugehörige Szene des Marienbegräbnisses: Eine Soldatengruppe verfolgt die Apostel, die Maria zu Grabe tragen. Petrus wendet sich um und stellt sich den Angreifern mit ausgestreckter rechter Hand entgegen. Die Juden, die versucht haben, sich des von vier Aposteln getragenen Sargs zu bemächtigen, sind zu Boden gestürzt, während ihre abgerissenen Hände am Sargtuch (schwarz mit weißem Kreuz) hängenblie ben. Johannes, dem der Wind Haar und Mantel verweht, trägt dem Zug einen hohen Palmzweig voran. Im Interkolumnium und auf dem oberen Seitenrand Rankentyp 3, auf dem rechten Rand Typ 2. Bl. 80vb Hl. Augustinus in prächtigem Bischofsornat, frontal auf einem wuchtigen weißen Thron sitzend, die Rechte segnend erhoben und von adorierenden Augustiner Chorherren flankiert. Die beiden vorderen Chorherren halten ihre schwarzen Kappen in den Händen, links der Propst mit grauem, schweifbesetztem Feh-Almutium. – Bas-de-page: Greifvogel stürzt sich (parallel begleitet von kleinerem Vogel im Sturzflug) auf Gans mit ihren Jungen herab, diesen voran eine Eidechse; Putto mit erhobenem Streitkolben greift von hinten Affen an, welcher aus einer Schale frisst, in der ein Putto herabgefallene Feigen sammelt. Diese werden von einem Putto in dem auf dem linken Seitenrand emporwachsenden Feigenast gepflückt, an den sich nach oben ein Blütenzweig (vgl. Bl. 7r) anschließt; darin zwei weitere Putti, von denen einer Flöte (?) spielt und trommelt, während der andere, zusammen mit einer Gans und einem Hasen (beide an den Beinen aufgehängt) kopfüber in den Ranken hängt. Auf dem oberen Seitenrand und im Interkolumnium Rankentyp 3; in Rankengabelung Eule, die blauen Vogel gefangen hat. Bl. 83rb Geburt Mariens: Vor Rundbogenfenster mit Ausblick in Berge und Himmel (dieser auf dem Außen grund fortgesetzt) sitzt Anna in einem Himmelbett (davor Kasten mit Zinnkanne und -becher) und löffelt aus einem Teller, während eine kniende Magd die kleine Maria (als Wickelkind) in einem Holzzuber badet. – Auf dem Bas-de-page Putti bei der Weinlese; mittig in den zarten Weinranken zwei weitere Putti, die großes Neustifter Wappen präsentieren (goldenes Tau-Kreuz auf rotem Grund, der von feinen Ranken in dunklerem Rot überzogen ist). Am rechten oberen Blattrand Rankentyp 3 mit Maikäfer, Marienkäfer und bräunlichem, schwarz geflecktem Schmetterling. In die Ranke größerer naturalistischer Zweig mit roten Blüten kopfüber eingehängt. Bl. 85ra Kirchenweihe: Bischof im Ornat mit goldenem Krummstab und Aspergil nähert sich, gefolgt von Klerikerschar (der vorderste mit Weihwasserkessel), einer Kirchenfassade mit schmalem Rundbogentor (mit Vordach, darüber Rundfenster; zudem Dachreiter erkennbar); über der Szene weht die Kirchweihfahne. – Als Randzier Rankentyp 3. Bl. 86va Allerheiligen: Heiligenschar; in der Frontreihe erkennbar (von links nach rechts): Katharina (Schwert), Barbara (Kelch und Hostie), Petrus (Schlüssel), Paulus (Schwert und Buch), Johannes (Kelch mit Schlange), Benedikt (schwarzer Habit). – Auf dem Bas-de-page großer Hirschkäfer und zwei aufgeschreckte, von bellendem Hund begleitete Putti, die wohl versucht haben, den Käfer mit einem Beil, jetzt am Boden liegend, zu attackieren. In der linken oberen Seitenecke roter Klee, an den ein auf den linken Rand herabhän gender Veilchenzweig mit einem roten Bändchen angebunden ist. Drolerie (Kleezweig): rosafarbener Falter. Bl. 88rb Totenmesse(n): Tisch mit schwarzem Tuch (mit weißem Kreuz), darauf Totenschädel und bren nende Kerze. – Rankentyp 3 (und 2) sowie kurze Blütenranke (vgl. Bl. 7r). Drolerie: Rabe mit Knochen im Schnabel. Bl. 58v Kussbildchen (55 × 180): Zwei prächtig gewandete Engel präsentieren das Schweißtuch mit dem dornenbekrönten Haupt Christi (mit delikatem Kreuznimbus). Bl. 1v ganzseitiges Stifterbild (250 × 200); interpretiert man das Buch in den Händen des hl. Augustinus als das Missale, dann auch Dedikationsbild: In schlichtem Goldrahmen (8 mm breit), mittig unter einem auf Marmorsäulen ruhenden und von Tondi flankierten Korbbogen, vor kobaltblauem Grund (mit feinlinigem Rautengitter), die Madonna in einer Mandorla (abwechselnd gerade und ondulierende Strahlen), mit einem Stab und dem Jesuskind auf dem Arm, das sich dem auf die Knie gesunkenen Auftraggeber der Hs., dem Neustifter Propst Augustin Posch, zuneigt; dieser trägt ein mit Schweifen besetztes Feh-Almutium, über ihm ein lebhaft gewundenes, leeres Spruchband; auf der anderen Seite der Madonna steht der hl. Augustinus im Bischofsornat mit kostbarem Krummstab und einem geöffneten Buch in den behandschuhten Händen; über der Madonna zwei Putti, die eine goldene Bügelkrone halten; zu Füßen der Gruppe drei weitere Putti als Wappenhalter: links Klostergründer Herren von Säben (auf silbernem Grund fünf aufsteigende rote Spitzen, mit feinen Ranken geschmückt), in der Mitte Neustift (auf rotem Grund goldenes Tau-Kreuz), rechts Augustin Posch (auf rotem Grund goldener Dreiberg, darüber stilisierter Ast mit zwei goldenen Früchten; Hermann 157: Erdbeeren). In der rechten Bildhälfte ein gut 40 mm breiter Streifen beschädigt. – Bas-depage: Auf einem vasenartigen Ziersockel sitzt ein Putto, der in zwei große Hörner bläst, denen Schwäne und Ranken (Typ 2) entwachsen. Zentral auf dem oberen Seitenrand halten zwei grün-karminrosafarbene Dra 124 Cod. 100 chen, die in Blattwerk übergehen, eine blaugrundige, goldgerahmte und von einer Fruchtschale bekrönte Kartusche mit der Jahreszahl 1526 (Goldziffern). Auf dem linken und dem rechten Seitenrand Kandelaber leisten, die rechte Leiste entwächst den Bas-de-page-Ranken; in der linken Leiste zwei Schwäne (an den henkelartigen Hälsen mit einem goldenen Band zusammengebunden und zugleich am zentralen Stab fixiert), zuoberst eine Fruchtschale mit drei Puttenköpfen. Bl. 51v ganzseitiges Kanonbild (245 × 195): dreifigurige Kreuzigung in einfachem Goldrahmen (50 mm breit). Im Boden verkeiltes Kreuz aus roh behauenen Baumstämmen, am Fuß umgeben von den Gebeinen Adams; der Gekreuzigte mit massiver Dornenkrone, beinah geschlossenen Lidern und rosafarbenem, ab flatterndem Lendentuch; drei Engel fangen das den Wundmalen Christi entströmende Blut in Kelchen auf, zwei der Engel schweben dabei unter den Querarmen des Kreuzes, der dritte kniet dahinter; Maria richtet, mit gefalteten Händen, den Blick verloren zu Boden, während Johannes mit leicht geöffnetem Mund und erhobenen Händen zum Gekreuzigten aufblickt. Minutiös gestalteter Hintergrund mit Stadtvedute vor alpinerBergkette, über der die Sonne an einem mit zarten Wolken überzogenen Himmel untergeht. In der rechten oberen Ecke Abdruck der Te-igitur-Initiale Bl. 52r. – Auf dem Bas-de-page drei zugehörige Passions szenen: Geißelung mit zwei Schergen, von denen einer Christus am Heiligenschein (!) festhält oder ihm in die Haare fassen will (Szene etwas abgerieben) und seinen Fuß fixierend auf das linke Bein Christi gestellt hat; die Geißelsäule zugleich der Sockel der linken Kandelaberleiste; daneben Dornenkrönung mit drei Schergen und Kreuztragung mit drei Soldaten, von denen einer Simon von Cyrene, der Jesus beim Tragen helfen will, an den Haaren festhält, um ihn mit einem Seil zu schlagen. Auf dem oberen Seitenrand edler Fruchtpokal, dem Ranken (Typ 2, mit Puttenköpfchen) entwachsen. Als Vorlage für Cod. 100 diente tw. Cod. 196 (zusammen mit dem Teil Esztergom, Kathedralbibl., Ms. II.2), vgl. z. B. die Augustinus-Initialen mit den adorierenden Chorherren (Bl. 80vb mit Cod. 196, Bl. 31[22]r; s. Hermann 163 Anm. 2) oder die Kirchenweihe-Initialen, deren Komposition im Posch-Missale seiten verkehrt übernommen wurde (Bl. 85ra mit Cod. 196, Bl. 34[25]r; s. Hermann 163 Anm. 3); darüber hinaus etliche Parallelen im Randschmuck, v. a. bei den naturalistischen Pflanzendarstellungen; zum einen Über nahme der Kombinationen zweier naturalistischer Zweige, wobei nur minimale Veränderungen vorgenom men wurden, vgl. Bl. 63v mit Cod. 196, Bl. 34[25]v (Akelei und Heckenrosen samt Vogel und rotem Falter) oder Bl. 45v mit Ms. II.2, Bl. 11v (Wiesensalbei und Butterblume samt Vogeldrolerie), des Weiteren Über nahme des naturalistischen Zweigs und Umwandlung des Akanthusasts zu Rankentyp 3, vgl. Bl. 39r mit Cod. 196, Bl. 32[23]r (Nelken samt Glucke, Akanthus mit zwei Vogeldrolerien), und schließlich wurde häufiger aus einer Kombination von Akanthus mit naturalistischem Zweig nur letzterer übernommen, vgl. die kopfüber auf dem Seitenrand platzierten Iriszweige Bl. 68v und Ms. II.2, Bl. 5v (sehr ähnliche Iris auch Cod. 196, Bl. 10v, hier mit der Blüte nach oben). Die Adaptationen sind nicht an die Textstelle und den Initialtyp gebunden, denen die jeweilige(n) Pflanze(n) in Cod. 196 bzw. Ms. II.2 zugeordnet wurde(n). Dies lässt an ein Musterbuch des 15. Jhs. denken, das in Neustift noch im 16. Jh. vorlag. Während die PflanzenIlluminationen quasi unverändert übernommen wurden, passte man die spätmittelalterlichen Akanthus ranken dem Renaissance-Geschmack an. – Zu den Vorbildern für die Gestaltung von Cod. 100 wurden viel fältige Überlegungen angestellt. H. Fiegl sieht die Ausstattung des Missales nur peripher durch die Donauschule und deren Naturauffassung beeinflusst, vielmehr durch die Motivik (v. a. des Randschmucks) und die Farbgebung der süddeutschen Buchmalerei des 1. Viertels 16. Jh. (Nürnberg und vorrangig Augs burg) sowie durch das Organisationsprinzip der Ausstattungselemente, das Albrecht Dürer für seinen Part der Ausschmückung im Gebetbuch Maximilians I. (1514 gedruckt, 1515 dekoriert) wählte (s. Fiegl, Missale 121–124; Fiegl, Prachthandschrift 752–754, passim), weshalb als führende Illuminatorenhand wohl ein süd deutscher Künstler angenommen werden kann, der nicht nur als Miniator, sondern auch als Tafelmaler tätig gewesen sein könnte (Fiegl, Missale 121). Eine weitere Inspirationsquelle dürften die verbreiteten Holz schnitte und Stiche süddeutscher Meister Anfang 16. Jh. gewesen sein (s. Hermann 163 Anm. 4). Während in der älteren Literatur noch Stephan Stetner als ausführender Künstler genannt wird (s. z. B. Hermann 156), spricht sich die neuere Forschung dagegen aus, da zumindest die Meisterhand des Cod. 100 eindeutig fort schrittlicher einzustufen ist als Stetners Arbeiten – immer vorausgesetzt, man nimmt diesen Neustifter Chor herrn in der Tat nicht nur als Schreiber, sondern auch als Buchmaler an. Eine Hypothese, die mit Vorbehalten zu sehen ist (hierzu vgl. Cod. 194). Fiegl, Prachthandschrift 748 kann sich zumindest eine untergeordnete Mitarbeit Stetners am Posch-Missale vorstellen, wobei sie Cod. 194 und 196 in ihre Überlegungen ein bezieht. S. R. E: Neuzeitlicher Bibliothekseinband: braunes Leder über Holz mit Blindlinien. Neustift, 2. H. 18. Jh. Einband und Bindung 2017 restauriert: gereinigt, Bindung und Heftung stabilisiert, Buchrücken gestärkt sowie Fehlstellen am Einbandrücken ergänzt (vgl. Restaurierprotokoll). Leder stark abgerieben, zerkratzt und beschädigt. VD und HD gleich: ein aus dreifachen Streicheisenlinien gebildeter Rahmen, zum Rücken Cod. 100 125 hin vertikal jeweils nochmals dreifache Streicheisenlinien. Reste von zwei Leder-Metall-Schließen. Kanten gerade und nach innen hin abgeschrägt. Rücken: Leder tw. 2017 ergänzt (deutlich erkennbar), fünf einfache Bünde, von (urspr. wohl Gold-)Bordüren gesäumt. Im zweiten Feld Golddruck MISSALE in Goldrahmung, im dritten Signaturschild (s. bei G). Kapitale mit naturfarbenem und grünem Spagat umstochen sowie mit Leder umschlagen. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Hs. von Stephan Stetner 1524 im Auftrag von Propst Augustin Posch (Propst 1519–1527) im Kloster Neu stift geschrieben, Buchmalerei wohl erst zwei Jahre später abgeschlossen (s. Bl. 1v: 1526). Zahlreiche Dar stellungen des Neustifter Wappens (Bl. 1v, 2r, 7r, 83r). Bl. 2r Besitzvermerk Ecclesiae Neocell. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD Signaturen der ULB Tirol II 2 G 4 (Bleistift) und Nr. 100 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 100. Bl. 1r und 2r Stempel der ULB Tirol, 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Triden tina 20. Jh. L: F. Leithe, Die k.k. Universitätsbibliothek in Innsbruck. Neuer Anzeiger für Bibliographie und Bibliotheks wissenschaft (1874) 260–266, hier 263. – Wilhelm II 110. – Hermann 155–164, Nr. 171. – Atz 1012. – B. Rutz, Illuminierte Choralhandschriften zu Neustift. Kirchenmusikalische Jahrbuch 22 (1909) 88–95, hier 90. – Gschwend 86. – Colophons Nr. 16895. – P. d’Ancona, E. Aeschlimann, Dictionnaire des miniaturistes. Mailand 1949, ND Nendeln 1969, 200. – I. Uhl, Buchmalerei. Geschichte, Technik und Ikonographie. Ettal 1971, 99–100 und Abb. 75. – T. Fabich, Die Buchmalerei der Frührenaissance in Süddeutschland. Wien 1972, 81–97, 149, Nr. 20. – W. Neuhauser, Eine unbekannte lateinische Beschwörungsformel in der sog. Neustifter-Innsbrucker-Spielehandschrift (Cod. 960 der Universitätsbibliothek Innsbruck), in: R. Muth, G. Pfohl (Hrsg.), Serta philologica Aenipontana 3 (Innsbrucker Beiträge zur Kulturwissenschaft 20). Innsbruck 1979, 221–253, hier 226 und 253. – Neuhauser, Neustift 85. – W. Salmen, Katalog der Bilder zur Musik geschichte in Österreich 1 (Innsbrucker Beiträge zur Musikwissenschaft IV). Innsbruck 1980, Nr. 585. – Katalog ULB Tirol I 279, 283–284. – Peintner, Buchmalerei 34–35, 93–115 (ohne Nennung der Sign.). – P. Spunar, Repertorium auctorum Bohemorum provectum idearum post Universitatem Pragensem conditam illustrans I (Studia Copernicana 25). Wratislaviae u. a. 1985, Nr. 120. – M. Peintner, Kloster Neustift. Au gustiner Chorherren in Südtirol. Bozen 1985, 19 (ohne Nennung der Sign.). – Peintner, Stiftsbibliothek 124–126. – M. Peintner, Das Skriptorium von Neustift, in: 850 Jahre Neustift. Katalog 91–93, hier 93 (ohne Nennung der Sign.). – Th. (H.) Innerhofer, Wappen und Siegel des Konvents und der Pröpste von Neustift, in: 850 Jahre Neustift. Katalog 199–214, hier 199 (ohne Nennung der Sign.). – Stefani 97. – Fiegl, Missa le. – Fiegl, Prachthandschrift. – U. Merkl, Buchmalerei in Bayern in der ersten Hälfte des 16. Jahrhunderts. Regensburg 1999, 22. – Neuhauser, Musikgeschichtliche Quellen 164, 194 (ohne Nennung der Sign.). – Peintner, Schreibkunst 371, Taf. 10–12 (ohne Nennung der Sign.). – Gozzi 548, Nr. 78. – M. Roland, Buch malerei, in: A. Rosenauer (Hrsg.), Geschichte der Bildenden Kunst in Österreich 3: Spätmittelalter und Re naissance. München u. a. 2003, 521–546, hier 534–535. – Th. (H.) Innerhofer, Das Augustiner-Chorherrenstift Neustift, in: H. Obermair, K. Brandstätter, E. Curzel (Hrsg.), Dom- und Kollegiatstifte in der Region Tirol – Südtirol – Trentino in Mittelalter und Neuzeit (Schlern-Schriften 329). Innsbruck 2006, 223–238, hier 228. – Roland, Renaissance 633, 635, 638, 643–644 (Kat. 334, tw. ohne Nennung der Sign.). – Baroffio 324, Nr. 16710–11. – Krämer, Scriptores, s. v. „Stetner, S.“. – D. Merlin, Due versioni melodiche del Pater Noster a Bressanone/Brixen ed una rara soluzione editoriale nel contesto dei messali a stampa all’inizio del Cinque cento, in: L. Rossi (Hrsg.), Ars Sacra 2013. Musica sacra e liturgia nelle Cappelle Musicali: testimonianze e testimoni. Giornate di studi musicologici, etnomusicologici e storici. Anagni, 21–24 marzo 2013. Roma 2017, 327–349. – U. Stampfer, Missale des Neustifter Propstes Augustin Posch, in: J. Kronbichler, H.-P. Ties (Hrsg.), Renaissance im Gebirge. Der Maler Bartlme Dill Riemenschneider und seine Zeit. Hofburg Brixen 27. Mai–31. Oktober 2017, 120–123 (Kat. 26). – J. Emprechtinger, Restaurierprotokoll, Mai 2017. Missale (festivum) für Propst Augustin Posch („Posch-Missale“). Mit Missale Brixinense (Druck: Basel 1511) weitgehend übereinstimmend, Anordnung abweichend: Ordo missae zw. Karsamstag und Ostersonntag eingeschoben. Rubriken bes. an den Kartagen aufgrund der Eigenliturgie tw. gegenüber Druck leicht abweichend. Vollständige Messformulare im Temporale nur für die Hochfeste, im Sanctorale für Marienfeste, Agnes, Augustinus, Allerheiligen und Kirchweihe, zusätzlich Sakramentar für zahlreiche Heiligenfeste. (1r) nur Rahmung. (1v) ganzseitige Miniatur (s. bei A). 126 Cod. 100 1 (2ra–36rb) Temporale, pars hiemalis (In nativitate dni bis In vigilia paschatis). (2ra) In vigilia nativitatis dni. – (3ra) In galli cantu. Mit Sequenz (AH 53, Nr. 10). – (5rb) In die sancto. Mit Sequenz (AH 53, Nr. 15). – (7ra) In circumcisione dni. Mit Sequenz (AH 53, Nr. 16). – (8rb) In epiphania dni. Mit Sequenz (AH 53, Nr. 29). – (9vb) In purificatione BMV. Mit Sequenz (AH 53, Nr. 99). – (11va) Dnca in palmis. – (17rb) Fer. 5 in coena dni. – (20va) Fer. 6 in parasceve. (28ra) Depositio (Gschwend 86). Inc.: Quid retribuam domino pro omnibus que retribuit mihi panem celestem accipiam de mensa domini … – (28va) Sabb. sanctus. Feuer- und Kerzenweihe mit anschließendem Exultet (im Druck nur Verweis). (29vb) Sechs Lectiones, jeweils mit anschließender Oratio: Lectio 1; Lectio 2 (im Druck nicht enthalten: Gn 22,1–19); Lectio 3 (im Druck: Lectio 2); Lectio 4 (im Druck: Lectio 3); Lectio 5 (im Druck: Lectio 4); Lectio 6 (im Druck nicht enthalten: Is 55,1–11). (34vb) Taufwasserweihe mit Orationen (mit Druck übereinstimmend) und anschließenden Gebeten, Rubriken und Hymnus unter ausdrücklic her Bezugnahme auf das Salzburger Missale: Sequitur secunda collecta continuando sub una conclusione secundum Salisburgensem. (35ra) Messe (mit Druck übereinstimmend). 2 2.1 (36va–61v) Ordo missae. (36va–38rb) Kyriale: Kyrie. Gloria. Credo. Agnus Dei. (36va) Kyrie (mit Notation). Gloria (einmal ohne, einmal mit Notation). – (36vb) Agnus Dei. – (37ra) Gloria mit Tropus Spiritus et alme. – (37rb) Intonation für zwei verschiedene Versionen des Credo. – (37vb) Gebete nach der Gabenbereitung. Darunter Datierung und Schreiber nennung: 1524 S S. 2.2 (38v–47v) Praefationes mit Notation. (38v) In die nativitatis domini prefatio. – (39r) De epiphania domini Ihesu prefatio solennis. – (40r) De sancta cruce sequitur prefatio dominicaliter. – (40v) In vigilia pasce prefatio de re surrectione dominicaliter sequitur. – (41v) In die sancto pasce prefatio de resurrectione domini solenniter. – (42r) Ascensionis domini prefatio solenniter. – (43r) In vigilia penthecostes prefatio dominicaliter de sanctis. – (43v) In die penthecostes prefatio solenniter. – (44v) De sancta trini tate prefatio solenniter. – (45v) De beata virgine prefatio solenniter. – (46v) Prefatio communis solenniter cantanda. – (47r) Prefatio communis ferialiter. 2.3 (47v) Forts. des Kyriale: Sanctus. Benedictus (beide ohne Notation). 2.4 (48ra–50rb) Communicantes. (48ra) De nativitate domini Ihesu infra actionem. – De epiphania domini infra actionem. – (48rb) In cena domini infra actionem. – (48vb) In vigilia pasce infra actionem. – (49rb) In die sancto pasce infra actionem. – (49vb) Ascensionis domini infra actionem. – In vigilia et in die sancto penthecostes. (50rb) Datierung: 1524. 2.5 (50v) Gebetschlüsse. (50v) Conclusio ultime collecte. Et famulos tuos antistitem regem et princepes (!) nostros cum omnibus sibi commissis … Per dominum nostrum Ihesum Christum filium tuum … vel sic Per eundem dominum nostrum … vel Qui cum Deo patre et spiritu sancto vivis … Oremus. Dominus vobiscum. Per omnia secula seculorum. Pax domini sit semper vobiscum. Darunter Datierung: 1524. (51r) leer. 2.6 (51v) ganzseitiges Kanonbild (s. bei A). 2.7 (52r–61r) Canon missae. Von Te igitur bis Doxologie. (56v) Pater noster, unterschieden nach Fest- und Werktagen, jeweils mit Notation. (57v) Libera nos. Pax vobiscum. (58r) Agnus Dei. Kommuniongebete (gegenüber Druck tw. abweichend). (61v) leer. Cod. 100 127 3 (62ra–73rb) Temporale, pars aestivalis (Dnca resurrectionis dni bis In festo ss. corporis Christi). (62ra) Dnca resurrectionis dni. Mit Sequenz (AH 53, Nr. 36). – (63va) In ascensione dni. Mit Sequenz (AH 53, Nr. 67). – (65rb) Sabb. in vigilia pentecostes. (65va) Vier Lectiones (tw. nur Verweis), jeweils mit anschließender Oratio (mit Druck übereinstimmend). (67rb) Taufwasser weihe (vom Druck leicht abweichend). (67va) Messe. – (68va) Dnca pentecostes et de spiritu sancto. Mit Sequenz (AH 53, Nr. 70). – (70rb) In festo ss. trinitatis. Mit Sequenz (AH 53, Nr. 81). – (71va) In festo ss. corporis Christi. Mit Sequenz (AH 50, Nr. 385). 4 (73rb–88rb) Sanctorale, pars hiemalis und pars aestivalis gemischt. (73rb) In conceptione immac. BMV. Mit Sequenz (AH 54, Nr. 188). – (74rb) Agnes. Mit Sequenz (AH 55, Nr. 51). – (76ra) In annuntiatione BMV. – (77rb) In visitatione BMV. Mit Sequenz (AH 54, Nr. 193). Oratio, Secreta und Postcommunio vom Druck abweichend. – (79rb) In assumptione BMV. Mit Sequenz (AH 53, Nr. 104). – (80va) Augustinus. Mit Sequenz (AH 54, Nr. 32). Lectio (Hbr 5,1–6) vom Druck abweichend. – (82va) Hermes. Nur Oratio, Secreta, Post communio. – (82vb) Adrianus. Wie im Druck nur Oratio, Secreta, Postcommunio. – Corbinianus. Wie im Druck nur Oratio, Secreta, Postcommunio. – (83ra) In nativitate BMV. Mit Sequenz (AH 53, Nr. 95). – (85ra) In dedicatione ecclesiae. Mit Sequenz (AH 53, Nr. 247). Postcommunio vom Druck abweichend. – (86rb) Omnes sancti. Mit Sequenz (AH 53, Nr. 112). Communio vom Druck abweichend. – (88ra) Caesarius. Nur Oratio, Secreta und Postcommunio. 5 (88rb–93rb) Missae pro defunctis. (88rb) Pro fidelibus defunctis in generali. Gradualteile. – (88vb) Pro defunctis episcopis vel prelatis. Gradualteile. – (89ra) Pro episcopo. Wie im Druck nur Sakramentarteile. – (89rb) Pro uno vel pluribus sacerdotibus. Wie im Druck nur Sakramentarteile. – (89va) In depositione unius. Sakramentarteile, nicht im Druck enthalten. – (89vb) Pro uno vel pluribus fundatoribus. Sakra mentarteile, nicht im Druck enthalten. – (90rb) In anniversario unius. Wie im Druck nur Sa kramentarteile. – (90rb) In anniversario plurium. Sakramentarteile, nicht im Druck enthalten. – (90vb) Pro benefactoribus. Sakramentarteile, nicht im Druck enthalten. – (91ra) Pro fratribus et sororibus. Wie im Druck nur Sakramentarteile. – (91va) Pro parentibus. Wie im Druck nur Sakramentarteile. – (91vb) Generalis. Anschließend Lectiones (Reihenfolge vom Druck abweichend): (92ra) 2 Mcc 12,43–46. – (92rb) Ez 37,12–14. – (92va) Apc 14,13. 1 Cor 15,20–23. (92va) Pro episcopis et prelatis (Reihenfolge vom Druck abweichend). Nur Lectiones: Ies. 58,10–11. Io 6,37–39. – (93ra) Io 6,53–54. Io 11,21–27. (93rb) Datierung und Schreibernennung: 1524 S: S: 6 6.1 (93rb–104ra) Sacramentarium. (93rb–97rb) Temporale. (93rb) Dnca 3. post epiph. dni. – (93va) Dnca 4. post epiph. dni. – (93vb) Dnca 5. post epiph. dni. – Dominica sexta. Im Druck nicht enthalten. – (94ra) Dnca in Septuages. – (94rb) Dnca in Sexages. – Dnca in Quinquages. (94va) Datierung und Schreibernennung: 1524 S: S: (94va) Dominica octava post trinitatis. – (94vb) Dominica nona. – (95ra) Dominica decima. – (95ra) Dominica undecima. – (95rb) Dominica duodecima. – (95va) Dominica tredecima. – (95vb) Dominica quartadecima. – (96ra) Dominica quindecima. Secreta und Postcommunio gegenüber Druck vertauscht. – (96ra) Dominica sedecima. – (96rb) Dominica decima septima. Secreta und Postcommunio vom Druck abweichend. – (96va) Dominica decima octava. – (96va) Dominica decima nona. – (96vb) Dominica vigesima. – (97ra) Dominica vigesima prima. – (97rb) Dominica vigesima secunda. 128 6.2 Cod. 106 (97va–104ra) Sanctorale. (97va) Philippus et Iacobus. – (97va) Sigismundus. Vom Druck abweichend. – (97vb) In inventione s. crucis. – (98ra) Alexander et Eventius. – (98rb) Monica. Im Druck nicht enthalten. – (98va) Florianus et socii. Vom Druck abweichend. – (98vb) Iohannes ante Portam Latinam. – (99ra) Gordianus et Epimachus. – (99rb) Pancratius. – (99va) Maria ad martyres. Im Druck nicht enthal ten. – (99vb) In translatione s. Albuini. – (100ra) Victor. Im Druck nicht enthalten. – (100rb) Urbanus papa. – (100va) Nicomedes. – (100vb) Marcellinus et Petrus. – (101ra) Erasmus. Secreta und Postcommunio vom Druck abweichend. – (101rb) Bonifatius et socii. Oratio und Postcom munio vom Druck abweichend. – (101va) Primus et Felicianus. – (101vb) Barnabas. Vom Druck abweichend. – (102ra) Basilides et socii. – (102rb) Vitus. – (102va) Marcus et Marcellianus. – (102va) Gervasius et Prothasius. – (102vb) Achatius et socii. Vom Druck abweichend. – (103ra) Ursula et undecim milia virginum. Oratio vom Druck abweichend. – (103rb) Severinus. Vom Druck abweichend. – (103va) Crispinus et Crispinianus. Oratio vom Druck abweichend. – (103vb) In translatione s. Amandi. Secreta und Postcommunio vom Druck abweichend. (104ra) Datierung und Schreibernennung: 1524 S. Stetner. (104rb) leer. (104v) nur Rahmung und Linierung. U. S. Cod. 106 IOHANNES IANUENSIS Perg. I, 110, I Bl. 365 × 285. Süddeutscher Raum (?), 1. H. 15. Jh. B: Haar- und Fleischseite deutlich unterscheidbar, wenige kleine Löcher und Nahtspuren. Lagen: (I–1)I (Vor satzbl., Pap. 18. Jh.) + 11.V110 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Ab Lage 5 (mit Ausnahme von Lage 9 und 11) am Lagenanfang Kustoden in arabischen Ziffern beginnend mit 7, d. h. vor Bl. 1 fehlen wohl zwei Lagen (Textverlust). Lagenreklamanten, tw. stark be- bzw. weggeschnitten. Nach Bl. 110 fehlt mindestens eine Lage (Bl. 110v Reklamante: unde postreme, Textverlust). Am rechten unteren Blattrand vereinzelt Zählung der einzelnen Blätter jeder Lage 1–10, tw. beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Spuren von Ledersignakeln, Bl. 68 Lederrest. S: Schriftraum 280/290 × 185/190. Zwei Spalten, von Tintenlinien gerahmt (seitlich von zweifachen), zu 43, meist 47–48 Zeilen. Zirkelstiche. Bastarda, Initien der einzelnen Abschnitte (jeweils ein bzw. zwei Zeilen) in Auszeichnungsschrift (Textualis formata [Textura]). Am Blattrand Notazeichen, Korrekturen und Ergän zungen, tw. von anderer Hand. A: Rote Auszeichnungsstriche und Unterstreichungen, rote und blaue Paragraphzeichen sowie einzeilige Lom barden. Bei Buchstabenwechsel (Bl. 4ra, 38va, 52vb und 68vb) jeweils mehrzeilige (30–65 mm) rote Initia le. Oberlängen in der ersten Zeile bisweilen mit cadellenartigen Ausläufern, Unterlängen in der letzten Zeile tw. gedehnt. Bl. 62v rote Zeigehand (beschnitten). Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. etwas abgerieben, bes. am unteren Rand. Am VD und HD Ecken mit braunem Leder verstärkt. Am HD Pappe an der Kante beschädigt. Rücken: braunes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck VOCABULARIUS in Goldrahmung, im dritten Si gnaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`3 […] (radiert, Tinte), II 2 C 10 (Bleistift) und Nr. 106 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 106. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den Cod. 107 129 italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller VI 122. – Stegmüller RB Nr. 4220 Suppl. – Katalog ULB Tirol II 30, 203. – Peintner, Stiftsbibliothek 122. (Ir–v) leer. (1ra–110vb) ‹Iohannes Ianuensis›: Catholicon sive Prosodia, Pars 5: Vocabularium, unvollständig: L (lucinia) – P (postremus) (Druck: GW 3182–3205). Inc. mut.: ] que vulgo dicitur Rothinholus. Oracius in sermone. Licinias (!) soliti inpensa (!) prandere coemptas. Lucinium -nii ubi solet candela lucere quia dat lumen et dicitur a luce … – Expl. mut.: … et comparatur postremus -or -simus [ (Ir–v) leer. A. P. / U. S. Cod. 107 COMPILATIONES ANTIQUAE I–III Perg. I, 81, I Bl. 355 × 230. Nordfrankreich (Raum Paris), zw. 1210 und 1234. B: Haar- und Fleischseite deutlich unterscheidbar, in regelmäßiger Abfolge gebunden. Nur sehr vereinzelt klei ne Löcher. Bl. 1–10 durchgehend kleines rundes (gestanztes) Loch. Bl. 5 und 6, 17–22 sowie 79–81 unteres Drittel weggeschnitten (ohne Textverlust), Bl. 80 auch letzte Zeile beschnitten. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 5.V50 + (V–1)59 + 2.V79 + 281 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 56 herausgeschnitten (Text verlust), zugehöriger Falz mit erhaltenen Repräsentanten jetzt zw. Bl. 55 und 56 (urspr. zw. Bl. 53 und 54). Bl. 80 und 81 eingehängte Einzelbl. Bl. 81 verkehrt, d. h. verso statt recto eingebunden. Am oberen Blatt rand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 205 × 110. Zwei Spalten, von Bleistiftlinien gerahmt, zu 51 Zeilen auf (weit über den Schrift raum hinausreichender) Bleistiftlinierung. Zirkelstiche. Littera Parisiensis. Aus inhaltlichen rechtshisto rischen Gründen wohl zw. 1210 und 1234 geschrieben. A: Rote Überschriften, nur tw. ausgeführt. Lombarden und Initialen nicht ausgeführt, kein Platz freigelassen (Ausnahme: Bl. 1ra Platz für Einleitungsinitiale freigelassen, spaltenbreit, elf Zeilen hoch). Am inneren Blattrand sowie links des Schriftraums Repräsentanten für Lombarden; am inneren Blattrand Repräsentan ten für Überschriften (quer zum Schriftraum). E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Am VD rechte obere Ecke eingerissen. Rücken: brau nes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck IUS CANONICUM in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Bl. 1r Besitzvermerk 15. Jh. Iste liber est monasterii sancte Marie vel ad Novamcellam. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 C 11 (Bleistift, über ältere radierte Signatur geschrieben) und N. 107 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 107. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 136. – Wretschko–Sprung 15. – Furtenbach–Kalb 20. – Neuhauser, Neustift 88. – Katalog ULB Tirol II 30, 203. 130 1 Cod. 123 (Ir–v) leer. (1ra–78va) ‹Bernardus Papiensis›: Breviarium Extravagantium sive Compilatio prima Decreta lium, unvollständig (Ed.: A. Friedberg, Quinque Compilationes antiquae. Leipzig 1882, ND Graz 1956, 1–65). (1ra–53vb) Lib. 1–4, tit. 3, cap. 3. Inc.: [Iuste iudicate] filii hominum et nolite iudicare secundum faciem sed secundum iustum iudicium iudicate ut ostendatis vero diligere iusticiam qui iudicatis … – Expl. mut.: … que dispen[ (Lib. 4, tit. 4 und 5, cap. 1 fehlt) (54ra–78va) Lib. 4, tit. 5, cap. 2–Lib. 5, tit. 37, cap. 14. Inc. mut.: ] facultas et manumittentis … – Expl.: … non enim potest esse pastoris excusatio si lupus oves comedit et pastor nescit. Schlussschrift: Expliciunt Decretales prime. (78vb) nur Rahmung und Linierung. Lit.: Schulte GQ I 78–82; Kuttner 322–344. 2 (79ra–80vb) ‹Iohannes Guallensis›: Compilatio secunda Decretalium, unvollständig (Ed.: A. Friedberg, Quinque Compilationes antiquae. Leipzig 1882, ND Graz 1956, 66–67). Tit.: De constitutionibus. – Inc.: [P]reterea de lege illa vel errore quem cives tuos asseris sta tuisse videlicet ut si quis teneret per XXX annos … – Expl. mut.: … cum ex quo electionis tue confirmationem accepisti [ (= Lib. 1, tit. 3, cap. 7) Lit.: Schulte GQ I 87–91; Kuttner 345–354. 3 (81ra–vb, verkehrt eingebunden, Text beginnend bei 81va) ‹Petrus Beneventanus Collivacinus›: Compilatio tertia Decretalium, unvollständig: Lib. 5, tit. 23, cap. 8–10 (Ed.: A. Friedberg, Quinque Compilationes antiquae. Leipzig 1882, ND Graz 1956, 134). (81va) Inc. mut.: ma]nifestam non que iudici nec que alii … – (81rb) Expl.: … que possint in abbatem assumi. Lit.: Schulte GQ I 87–91; Kuttner 355–368. (Ir–v) leer. P. K. Cod. 123 BERNARDUS DE GORDONIO. MEDIZINISCHE TRAKTATE Pap. IV, 227, I Bl. 315 × 215. Süddeutscher Raum, 6. Jz. 15. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 586 (dat. 1459); Neustift, Stiftsbibl., Cod. 125; Brixen, Priesterseminarbibl., Cod. R.3. Urspr. Lagenformel aufgrund der Neubindung im 20. Jh. (s. bei G) nicht mehr eindeutig nachvollziehbar, urspr. wohl Sexternionen: Bl. 21v, 33v, 43v, 55v, 76v, 87v, 98v, 110v, 133v und 144v Lagenreklamanten erhalten. Gegenbl. zum Vor- und Nachsatzbl. (20. Jh.) am Spiegel des VD bzw. HD aufgeklebt. Bl. 1–166 am oberen Blattrand der Rectos zeitgenössische Foliierung in arabischen Ziffern (Register und leere Blätter ungezählt), Bl. 155, 157 und 158 fehlen (Textverlust). Ab Bl. 167 Forts. der Foliierung 19. Jh. durch die ULB Tirol. Mehrere Blätter in der Zählung (1–220) nicht berücksichtigt, später nachgetragen als 11a, 35a–b, 64a–c, 95a, 121a und 139a. S: Vorsatzlage (Bl. IIr–Vr) Schriftraum 255 × 160. Zwei Spalten, seitlich von Tintenlinien begrenzt, zu 66 Zei len. Bl. 1r–220r Schriftraum wechselnd 210/240 × 135/140, tw. von Tinten-, tw. von Bleistiftlinien gerahmt, zu 39–58 Zeilen. Bastarda von zwei Händen: 1) Bl. 1r–211r und 213r–220r; 2) Bl. IVv–Vr, 11av, 18v, 212r– v sowie zahlreiche Nachträge und Ergänzungen am Blattrand (= späterer Duktus des Schreibers von Neu stift, Stiftsbibl., Cod. 453, T. II–IV und Cod. 748, T. I–III, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Cod. 123 131 A: Rote Auszeichnungsstriche, vereinzelt Paragraphzeichen, Durchstreichungen, Unterstreichungen und Über schriften. Bl. 1v–10v zwei- bis vierzeilige, Bl. 32v–33v vier- bis siebenzeilige, Bl. 159r–166r drei- bzw. vierzeilige rote Lombarden. Bl. Vv und 139v Neustifter Wappen in schwarzer Tinte. E: Gotischer Einband: dunkelbraunes Leder über Holz mit Blinddruck. Tirol (?), 2. H. 15. Jh. Einband restauriert bzw. unter Verwendung des urspr. Lederbezugs am VD und HD im 20. Jh. erneuert. Urspr. Leder tw. stark abgerieben und zerkratzt. VD und HD gleich: zwei aus je zweifachen Streicheisen linien gebildete Rahmen. Äußerer Rahmen leer, Mittelfeld durch zweifache Streicheisenlinien diagonal in mehrere Rautenfelder unterteilt; darin jeweils ein kreisförmiger Stempel (Nr. 1). Spuren von zwei Kurzrie menschließen. Kanten gerade. Rücken: Leder erneuert, fünf einfache Bünde. Im ersten Feld Signaturschild (s. bei G). Kapitale mit hellblauem und rotem Zwirn umstochen. Spiegel sowie je ein Vor- und Nachsatzbl. 20. Jh. Am Spiegel des VD zwei Fragmente (wohl Teile des urspr. Spiegels) eingeklebt: 1) mit Besitz vermerk (s. bei G); 2) mit Spruch Seneca. Nichil est homini bonum sine se bono. Am Spiegel des HD Etikett der Restaurierwerkstätte (s. bei G). G: Geschenk des Neustifter Präbendars und Klosterarztes Bertoldus de Saxonia ans Stift (gemeinsam mit Cod. 748 und vermutlich auch dem tw. von denselben Händen geschriebenen Cod. 453): Bl. Vv Schen kungsvermerk (s. Nr. 2) mit Neustifter Wappen. Zu Bertoldus vgl. Liber anniversariorum Novae Cellae Brixinensis (MGH Necrologia Germaniae III 46): „Item magister Bertoldus de Saxonia, medicine doctor, prebendarius et phisicus huius monasterii, in vita sua multa fecit bona et libere omnia sua dedit monasterio, prout continetur in litera prebendali, quam monasterio dedit“. Besitzvermerke 15. Jh.: Am Spiegel des VD Iste liber est monasterii sancte Marie virginis in Novacella prope Brixinam, Bl. 139v Iste liber est monasterii sancte Marie virginis in Novacella mit Neustifter Wappen. Bl. 1r Besitzvermerk 18. Jh. Collegii Neocellensis. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro, S. Maria di Rosano [Firenze]), kein Restaurierbericht erhalten. Bl. Ir Signatur Cod. 123 (Bleistift). Am Rücken Neustifter Signaturschild 20. Jh. 123. L: Wilhelm II 110. – Neuhauser, Neustift 89. – Katalog ULB Tirol II 72, 203. (Ir–v) leer bis auf Signatur (s. bei G). (IIr–IVr) nur Rahmung. (IVva–Vra) Inhaltsverzeichnis zu Nr. 3. (Vrb) leer. 1 (Vv) Schenkungsvermerk mit Inhaltsangabe: Liber iste legatus est monasterio sancte Marievir ginis ad gracias alias ad Novamcellam communiter appellati (!) a felicis recordacionis magistro Bertoldo de Saxonia arcium et medicine doctore eximio cum aliis libris et rebus pro prebenda ac remedio anime sue. Obiit 1476. Liber iste continet Lilium medicine editum a magistro Bernardo de Gordonio. Tractatum de febribus. Item alium de asmate. 2 (1r–166r) Bernardus de Gordonio: Practica seu Lilium medicinae, unvollständig (Druck: GW 4080–4084. – Thorndike–Kibre 771–772. L. E. Demaitre, Doctor Bernard de Gordon: Professor and practitioner [Pontifical Institute of Medieval Studies, Studies and Texts 51]. Toronto 1980, 185–188). Gegenüber Druck tw. leicht abgeändert, tw. gekürzt. (1r) Hinweis am oberen Blattrand 18. Jh.: Bernardus de Gordonio. De febribus et asmate. (1r) Tit.: Incipit liber qui dicitur Lilium medicine editus a magistro Bernardo de Gordonio in preclarissimo studio montis Pessulani. (1r) Prolog. Inc.: [I]nterrogatus Socrates a quodam quomodo posset optime respondere seu di cere. Respondit Si nichil … – Expl.: … post annum XX lecture nostre anno domini Mo CCCo IIIo mense Iulii. (1r) Kapitelübersicht zu Lib. 1. – (1v) Lib. 1, cap. 1–10. (11v) nur Rahmung. 132 Cod. 123 (11ar) Nachtrag zu Lib. 1, cap. 10, durchgestrichen. Inc. mut.: ] non omnino sunt certe pro nuncciaciones neque vite neque mortis. Curacio rectificetur aer … – Expl. mut.: … de corde cervi 3. 1 misceantur [ Am oberen Blattrand Notiz von anderer Hand: Nichil vacat. (11av) Ergänzung zu cap. 11 (einzufügen Bl. 12v, Z. 5). Inc.: Curacio. Aut enim intendimus sudorem provocare aut auferre … – Expl.: … dyafori sicud narrat Galienus etc. (12r) Lib. 1, cap. 11–31. (18v) leer bis auf Nachtrag zu Lib. 1, cap. 18. (35ar–35br) nur Rahmung. (35bv) Kapitelübersicht zu Lib. 2. – (36r) Lib. 2. (64v–64cr) nur Rahmung. (64cv) Kapitelübersicht zu Lib. 3. – (65r) Lib. 3. (83av) Kapitelübersicht zu Lib. 4. – (84r) Lib. 4, cap. 1–5 sowie 11–13 (in der Hs. als 10–12 angeführt). (95v–95ar) nur Rahmung. (95av) Kapitelübersicht zu Lib. 5. – (96r) Lib. 5. (121v–121ar) nur Rahmung. (121av) Kapitelübersicht zu Lib. 6. – (122r) Lib. 6. (139ar) nur Rahmung. (139av) Kapitelübersicht zu Lib. 7. – (140r) Lib.7, cap. 1–20, unvollständig. (154v) Expl. mut.: … et si corpus est mundificatum tanto melius etc. [ (Textverlust: Bl. 155 fehlt, s. bei B). (156r–v) Cap. 21, unvollständig, bricht nach dem vorletzten Rezept ab. Expl. mut.: … vel conficiantur cum melle et zuccharo (andere Tinte:) etc. [ (Textverlust: Bl. 157–158 fehlen, s. bei B) (159r–166r) Lib. 4, cap. 6–10. Lit.: L. E. Demaitre, Doctor Bernard de Gordon: Professor and practitioner (Pontifical Institute of Medieval Studies, Studies and Texts 51). Toronto 1980. (166v–170v) nur Rahmung. (171r–194v) leer. 3 (195r–211r) Tractatus de febribus. (195r–202r) Inc.: [F]ebris secundum Avicennam quarto canonis in principio est calor extraneus accensus in corde et ab eo procedens mediantibus spiritu et sanguine … Et talis calor extraneus quandoque est in spiritibus, quandoque in humoribus … – Expl.: … sed est magis tristis et magis timidus. (202v) nur Rahmung. (203r–211r) Inc.: [C]um de febribus flegmaticis iam tractandum est. Primo videndum est … – Expl.: … et sparagi. Et hec de cura flegmatice, a flegmate pontico provenientis. Laus Deo. (211v) leer. 4 (212r–215r) Tractatus de asmate. Tit.: De asmate. – Inc.: Asma ut dicit Avicenna est egritudo pulmonis cum qua paciens non inve nit excusacionem ab anhelitu … Et Rasis addit tussim et quod leditur ex iacendo … – Expl.: … ficubus siccis pressis et ysopo et ca. ve. etc. 5 (215v–216v) Nachtrag zu Bernardus de Gordonio: Practica seu Lilium medicinae, Lib. 3, cap. 16. Inc.: Reuma est fluxus humorum ad partes subiectas. Reumatis autem tres species ab autoribus distinguntur … – Expl.: … in passionibus pectoralum ad preservandum ne materia vadat ad pectus. (217r–v) nur Rahmung. Cod. 125 6 133 (218r–220r) Nachtrag zu Bernardus de Gordonio: Practica seu Lilium medicinae, Lib. 4, cap. 4, unvollständig: Text bricht in der Mitte der dritten Rubrica ab. Verweis am Blattrand: Idem capitulum habetur 87. – Inc.: [T]ussis est motus naturalis ex pul mone … – Expl.: … simpliciter tamen loquendo tussis est motus quoniam ledit operationes aut significat lesionem cum concutiatur. (220v) leer. (Ir–v) leer. L. Š. Cod. 125 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. I, 313, I Bl. 310 × 220. Süddeutscher Raum, 6. Jz. 15. Jh. B: Bl. 1 stark beschädigt, Fehlstellen bzw. Risse mit Pap. geklebt. Bl. 310 Loch (Textverlust), Bl. 311 loses Einzelbl., stark beschädigt (Textverlust). Nachweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Neustift, Stiftsbibl., Cod. 123; Innsbruck, ULB Tirol, Cod. 758 (dat. 1455). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 26.VI310(312) + 1311(313) + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. I und 1 etwas kleiner als restlicher Buchblock (310 × 210). In der Mitte der ersten und der letzten Lage unbeschriebener Pergamentfalz. Lagenreklamanten, Lagen 1–13 meist stark beschnitten, ab Lage 14 (Bl. 168v) gänzlich erhalten. Am oberen Blattrand Foliierung 19. Jh.; Fehler in der Zählung: 271 und 288 doppelt angeführt. Bl. 98, 125 und 205 jeweils neuzeitliches oranges Papiersignakel der ULB Tirol. S: Schriftraum 210/230 × 130/145. Zwei Spalten, bis Bl. 24v vorwiegend von Blindlinien, anschließend von Tintenlinien gerahmt, mit wechselnder Zeilenzahl. Zirkelstiche. Bastarda von mehreren Händen: 1) Bl. 1ra– 97va (zu 34–40 Zeilen); 2) Bl. 98ra–125ra (zu 35–37 Zeilen); 3) Bl. 125va–160vb (zu 38–43 Zeilen); 4) Bl. 161ra–200ra (zu 50–54 Zeilen); 5) Bl. 205ra–311v (zu 44–48 Zeilen). A: Rote Auszeichnungsstriche, Paragraphzeichen, Zeilenfüllsel, Unterstreichungen sowie Über- und Schluss schriften. Rote und schwarze Zeigehände. Ein- bis fünfzeilige rote Lombarden, tw. mit Punktverdickungen, tw. mit einfachen Ausläufern, sehr vereinzelt mit einfachem Knospenfleuronnée in Schwarz. Bl. 1ra fünfzei lige rote Initiale mit einfachem Knospenfleuronnée im Binnenfeld, als Besatz lange, orthogonal abstehende Fadenausläufergruppen. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, tw. beschädigt, vier Doppel bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck KAMPIGALL. (!) BIBLIA AUREA in Goldrahmung, im dritten Signaturschild (s. bei G). Kapitale mit naturfarbenem Spagat umstochen. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`4 D 6 G 6 (? radiert und durchgestrichen, Tinte), II 2 E 12 (Bleistift) und Nr. 125/3 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signatur schild der ULB Tirol 125/(3). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleih gabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kaeppeli Nr. 1622. – Stegmüller RB Nr. 124,1.1 Suppl. – Zumkeller Nr. 115. – K. Walsh, Von der scholasti schen Literatur zur Pastoraltheologie. Die „Augustinerschule“ im Spiegel der Stamser Handschriften, in: Studia Stamsensia (Innsbrucker historische Studien 6). Innsbruck 1984, 27–44, hier 40 Anm. 62. – Katalog ULB Tirol II 75, 204. – Peintner, Stiftsbibliothek 121. – D. E. Mairhofer, Liber Lacteus. Eine unbeachtete Mirakel- und Exempelsammlung aus dem Zisterzienserkloster Stams (Codicologia 1). Badenweiler 2009, 23 Anm. 33. 134 1 Cod. 125 (Ir–v) leer. (1ra–97va) Antonius Rampegolus: Biblia aurea (Druck: u. a. GW M36970. – Stegmüller RB Nr. 1419. Zumkeller Nr. 115). (1r) Tit.: Incipit Biblia aurea sancti alphabeti (ergänzt: author Antonius Kampigallus [!] ordinis Eremitarum sancti Augustini). (1ra) Prolog. Inc.: Religiosis atque honestis viris in Christo dilectis fratribus studentibus Nea polim … – Expl.: … consuevit egredi a sua (!) misericordie largitate Amen. (1va) Register. Tit.: Tabula capitulorum. (2va) Alphabetisches Stichwortverzeichnis. Tit.: Item alia tabula. (9ra) Text. Inc.: Abstinencia est meriti augmentativa. Sapiencie acquisitiva … – Expl.: … accepit epistolas ut persequeretur Christianos Act. 13. Deo gracias. Genaue Aufschlüsselung des Inhalts s. bei Stegmüller. (97vb) leer. 2 (98ra–125rb) Exzerpte aus ‹Guilelmus Peraldus›: Summa de vitiis et virtutibus, De virtutibus (Druck: u. a. GW 12055. – Kaeppeli Nr. 1622B. Bloomfield Nr. 5601). (98ra) Register. Tit.: Summa virtutum. (98rb) Text. – (125rb) Schlussschrift: Expliciunt excerpta de summa virtutum. Deo gracias. 3 (125va–145vb) ‹Gerardus de Vliederhoven›: Cordiale quattuor novissimorum (Druck: u. a. GW 7473. – Madre 329. Bloomfield Nr. 3057). Tit.: Sequitur Cordiale tractatus de quatuor novissimis optima materia. Auch zugeschrieben Henricus de Hassia, Iohannes Gerson, Iacobus Gruytroede, Dionysius Car thusianus. 4 (146ra–160vb) ‹Thomas a Kempis›: De imitatione Christi (Ed.: J. Pohl, Thomae Hemerken a Kempis … opera omnia II. Friburgi Br. 1904. – Bloomfield Nr. 4633). Abweichende Kapitelzählung in der Hs., da einige Kapitel nicht gezählt, vereinzelt Kapitel gekürzt bzw. ausgelassen, s. Lib. 3. (146ra–151ra, Z. 11) Lib. 1. – (151ra, Z. 11–154va, Z. 14) Lib. 2, beginnend mit cap. 1.2 ohne Hervorhebung (Eya anima fidelis prepara …). – (154va, Z. 15–159ra, Z. 34) Lib. 3, cap. 1–59.3. – (159ra, Z. 35–160vb) Lib. 4. 5 (161ra–169va) ‹Iacobus Mediolanensis› = ‹Ps.-Bonaventura›: Stimulus amoris, Auszug (Druck: A. Ch. Peltier, S. R. E. Cardinalis S. Bonaventurae … opera omnia XII. Parisiis 1868, 633–703. Stimulus amoris fr. Iacobi Mediolanensis. Canticum pauperis Fr. Ioannis Peckam [Bibliotheca Franciscana Medii Aevi 4]. Quaracchi 21949, 1–129. – Distelbrink Nr. 217–219. Bloomfield Nr. 1294). Die Hs. enthält Pars 1, cap. 1–4; Pars 2, cap. 1, 6, 9, 10; Pars 3, cap. 1–5 des Drucks (1868). Rubrik: Liber ille (!) qui intitulatur Stimulus amoris in dulcissimum et piissimum salvatorem nostrum in tres dividitur partes … – Inc.: Currite gentes undique et miramini erga vos caritatem Dei … – Expl.: … misericors est Deus qui est benedictus et laudabilis et gloriosus in secula seculorum Amen. Lit.: F. Eisermann, „Stimulus amoris“. Inhalt, lateinische Überlieferung, deutsche Übersetzun gen, Rezeption (MTU 118). Tübingen 2001. Vgl. Cod. 165, Bl. 132v. 6 (169va–170ra) Auszug aus ‹Ecbertus Schonaugiensis›: Soliloquium seu Meditationes, cap. 1–4 (PL 195, 105D–108B). Tit.: Contemplacio Bernhardi. Vgl. Cod. 165, Bl. 139v. Cod. 125 135 7 (170rb–183vb) ‹Ps.-›Eusebius Caesariensis: Epistola ad beatum Damasum episcopum Portuen sem et ad christianissimum Theodosium Romanorum senatorem de morte Hieronymi (PL 22, 239–282. Klapper II 10–241. – BHL Nr. 3866. BHM Nr. 903E). Abweichende Kapiteleinteilung. Tit.: Incipit libellus de vita et obitu et miraculis sancti Ieronimi et primo epistula Eusebii ad Damasium (!) et Theodonium (!) Romanorum imperatorem senatorem. Schlussschrift: Explicit epistula Eusebii de morte sanctissimi Ieronimi presbyteri. 8 (183vb–186va) ‹Ps.-›Augustinus: Epistola 18 (ad Cyrillum Hierosolymitanum episcopum de magnificentiis beati Hieronymi) (PL 22, 281–289. Klapper II 245–288. – CPL Nr. 367. BHL Nr. 3867. BHM Nr. 903A). Tit.: Incipit epistula beati Augustini ad Cyrillum patriarchum Ierosolimitanum de beatissimo Ieronimo. Exordium epistolare de non tacendo laudes Ieronimi. Schlussschrift: Explicit epistula Augustini ad Cirillum de glorioso Ieronimo. 9 (186va–198va) ‹Ps.-›Cyrillus: Epistola ad Augustinum de miraculis Hieronymi (PL 22, 289– 326. – CPL Nr. 367. BHL Nr. 3868. BHM Nr. 903C) = ‹Ps.-Augustinus›: Epistola App. 19 (PL 33, 1126–1153). Tit.: Incipit epistula Cirilli ad Augustinum de miraculis gloriosi Iheronimi. 10 (198va–200ra) Auszüge aus ‹Iohannes Andreae›: Hieronymianus (Druck: GW 1727. – BHL Nr. 3876. BHM Nr. 907). (198va) Tit.: De quibus miraculis factis noviter in Appulia et primo quid in Troya civitate. – Inc.: Sciendum est quod moderno tempore in Appulia in provincia Capitinate … – Expl.: … et nota biliter fide digni. (198vb) Tit.: De magistro Iacobo liberato. – Inc.: Magister Iacobus scutellarius commendabilis vite … – Expl.: … subsidium obtulit. (199ra) Tit.: De mortuo resuscitato. – Inc.: Item magister barbatus sancti Georii … – Expl.: … et aduc (!) vivit. (199ra) Tit.: De capto liberato. – Inc.: Item dum exercitus domini regis Roberti … – Expl.: … votum implevit. (199rb) Tit.: De liberato ab aquis. – Inc.: Item Favinus de Slappis … – Expl.: … elevaverunt eundem. (199rb) Tit.: De liberato a casu. – Inc.: Item cum quidam frater in subscripta manens … – Expl.: … illesus evasit. (199rb) Tit.: De abortiva. – Inc.: Item quedam mulier in Arriano peperit … – Expl.: … puella revixit. (199rb) Tit.: De falcone invento. – Inc.: Item quidam nobilis falconem unum perdiderat … – Expl.: … falco venit. (199rb) Tit.: De sanacione equi. – Inc.: Item miles quidam Gallicus familiaris … – Expl.: … votum replevit sancto Ieronimo gracias agens. Die letzten beiden Auszüge in Wilten, Stiftsbibl., Cod. 32 02 05 zu einem Kapitel zusammen gefasst. (199rb) Tit.: De liberacione unius pueri. – Inc.: Item Petrus frater domini Mathei de Sallerno … – Expl.: … in eterna beati Ieronimi presentatus. (199va) Tit.: De ceco illuminato. – Inc.: Nicolaus Iohannis mariscalchi (!) … – Expl.: … restitutus est visus. (199va) Inc.: Item maiora (wie Wilten, Stiftsbibl., Cod. 32 02 05, Bl. 427rb, im Druck „Maria“) de terra … – Expl.: … reliqui operando. (199va) Tit.: De puero liberato a flammis. – Inc.: Item Iohannes infans cum sorore maiore … – Expl.: … fuit illesus. (199va) Inc.: De duabus ecclesiis noviter … – Expl.: … montis Deglin (?). 136 Cod. 125 (199va) Tit.: De ecclesia sub terra cooperta. – Inc.: Deserta (in Wilten, Stiftsbibl., Cod. 32 02 05, Bl. 427va: „De secunda“) cui inest miraculum … – Expl.: … concursum civium festinatur. (199vb) Tit.: Quomodo translatum est corpus suum Romam. – Inc.: In quadam legenda transmis sa de Roma … – Expl.: … et devotissime laudavit. (200ra) Tit.: De blasphemo privato. – Inc.: Hereticus quidam illud gloriosum corpus dicens publice … – Expl.: … multi heretici ad lumen fidei et gremium ecclesie sunt reversi. (200rb–204v) nur Rahmung. Bis auf den letzten Text dieser Hs. dieselbe Auswahl der Kapitel auch in Wilten, Stiftsbibl., Cod. 32 02 05, vereinzelt Überschriften, sofern vorhanden, geringfügig abweichend. Nr. 7–10 Texte in dieser Anordnung auch in Cod. 165, Bl. 42va–74rb. Lit.: Schulte GQ II 217. 11 (205ra–311vb) Sibylla Tiburtina (= Liber miraculorum dictus Lacteus) (Bloomfield Nr. 1447, 3580. Stegmüller RB Nr. 124,1 und Nr. 124,1.1 Suppl.). Ein Großteil der Exempla ist in der Legenda Aurea (hier abgeglichen mit Ed. Graesse; zuletzt ed. von G. P. Maggioni, Iacopo da Varazze. Legenda Aurea [Millenio Medievale 6, Testi 3]. Tavarnuzze [Firenze] 1998) enthalten. Die vorliegende Sammlung stimmt mit München, BSB, Clm 23420 überein; sämtliche Exempla sind, wenn auch tw. anders angeordnet, in Innsbruck, ULB Tirol, Cod. 494 (Mairhofer, Liber Lacteus) enthalten. Zahlreiche Exempla auch in Neustift, Stiftsbibl., Cod. 308. Die Titel der einzelnen Exempla werden nur dann zitiert, wenn sie bei Mairhofer nicht aufschei nen oder von den dort angeführten Titeln abweichen. Text bisweilen von Editionen und Drucken abweichend, aber mit Mairhofer übereinstimmend. Inc. und Expl. werden nur dann angeführt, wenn sowohl von Editionen und Drucken als auch von Mairhofer abweichend. Tit.: Incipit liber miraculorum dictus Lacteus qui dividitur in novem distinctiones. 11.1 (205ra–215ra) Dist. 1: Miracula de Iesu Christo. (205ra) Inhaltsübersicht. (205rb) Tit.: Quod Augustus dictus Octavianus noluit se primum adorare pro Deo et visiones Sibille de nativitate Christi (Graesse Nr. 6 [S. 44, Z. 11–31]. – Mairhofer Nr. 1). (205rb) Graesse Nr. 6 (S. 42, Z. 1–9). – Mairhofer Nr. 2. (205va) Graesse Nr. 6 (S. 43, Z. 31–S. 44, Z. 4). – Mairhofer Nr. 3. (205va) Zwei Teile: Tit. 1: Quod omnes sodomite perierunt (Graesse Nr. 6 [S. 44, Z. 4–8]). Tit. 2: Idem (Graesse Nr. 6 [S. 45, Z. 34–S. 46, Z. 1]). – Mairhofer Nr. 4. (205va) Tit.: De eodem (Graesse Nr. 6 [S. 44, Z. 32–S. 45, Z. 2]. – Mairhofer Nr. 5). (205vb) Tit.: De Maria et Ioseph et asino (Graesse Nr. 6 [S. 45, Z. 8–15]. – Mairhofer Nr. 6). (205vb) Tit.: De Hugone abbate (Graesse Nr. 6 [S. 46, Z. 14–26]. – Mairhofer Nr. 7). (205vb) Tit.: De crucifixo et quodam peccatore (Graesse Nr. 6 [S. 46, Z. 26–33]. – Mairhofer Nr. 8). (206ra) Tit.: De circumcisione et nomine Ihesu (Mairhofer Nr. 9). (206rb) Mairhofer Nr. 10. (206va) Mairhofer Nr. 11. (206vb) Mairhofer Nr. 12. (207ra) Graesse Nr. 137 (130), 5. – Mairhofer Nr. 245. (207rb) Tit.: De virtutibus sancte crucis (Mairhofer Nr. 13). (207va) Graesse Nr. 137 (130), 3. – Mair hofer Nr. 14. (207vb) Graesse Nr. 137 (130), 4 (S. 608, Z. 26–S. 609, Z. 18). – Mairhofer Nr. 15. (208ra) Mairhofer Nr. 16. (208rb) Mairhofer Nr. 17. (208va) Mairhofer Nr. 18. Mairhofer Nr. 19. (209rb) Mairhofer Nr. 20. (209va) Graesse Nr. 100 (95) (S. 430, Z. 18–S. 431, Z. 31). – Mair hofer Nr. 21. (210ra) Graesse Nr. 100 (95) (S. 431, Z. 31–S.432, Z. 32). – Mairhofer Nr. 22. (210va) Mairhofer Nr. 23. (210vb) Graesse Nr. 68 (64) (S. 303, Z. 30–S. 304, Z. 7,9–20,22– 32). – Mairhofer Nr. 24. (211ra) Mairhofer Nr. 246. (211ra) Tit.: De iuranti per se crucem (Mair hofer Nr. 247). (211rb) Mairhofer Nr. 25. (211rb) Graesse Nr. 54 (52) (S. 242, Z. 3–32). – Mair hofer Nr. 26. (211va) Mairhofer Nr. 27. (212va) Mairhofer Nr. 28. (212va) Graesse Nr. 181 (176), 4 (S. 838, Z. 25–34). – Mairhofer Nr. 29. (212vb) Mairhofer Nr. 30. (213ra) Tit.: De milite qui sancta loca visitavit (Mairhofer Nr. 34). (213rb) Mairhofer Nr. 31. (213va) Tit.: Quod dyabolus bella suscitat (Graesse Nr. 181 [176], 4 [S. 842, Z. 15–24]. – Mairhofer Nr. 35). (213va) Cod. 125 137 ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 192 (PL 179, 1172A–B. – Mairhofer Nr. 36). (213vb) Tit.: De sancta trinitate et tribus guttis et gemma (Graesse Nr. 181 [176], 1 [S. 824, Z. 31–S. 825, Z. 7]. – Mairhofer Nr. 37). (214ra) Mairhofer Nr. 32. 11.2 (215ra–222ra) Dist. 2: Miracula BMV. (215ra) Tit.: Incipit secunda distinccio in qua agitur de miraculis sancte Marie et continet XXX miracula. (215ra) Inhaltsübersicht. (215rb) Tit. (zum auf den Prolog folgenden Exemplum): De virtute cuiusdam cerci (!) in purifi cacione sancte Marie. Prolog (Mairhofer Nr. 1.2, S. 91), direkt im Anschluss Graesse Nr. 37, 2. – Mairhofer Nr. 38. (215vb) Graesse Nr. 37, 3. – Mairhofer Nr. 39. (215vb) Graesse Nr. 119 (114), 6. – Mairhofer Nr. 40. (215vb) Graesse Nr. 119 (114), 8. – Mairhofer Nr. 41. (216ra) Graesse Nr. 119 (114), 2. – Mairhofer Nr. 42. (216ra) Graesse Nr. 131 (126), 2. – Mairhofer Nr. 43. (216ra) Graesse Nr. 88 (83) (S. 367, Z. 18–S. 368, Z. 2). – Mairhofer Nr. 44. (216rb) Graesse Nr. 131 (126), 3. – Mairhofer Nr. 45. (216rb) Mairhofer Nr. 46. (216va) Graesse Nr. 131 (126), 8. – Mairhofer Nr. 47. (216va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 14 (PL 185, 1077D–1078B. – Mairhofer Nr. 48). (216vb) Graesse Nr. 131 (126) (S. 589, Z. 39–S. 590, Z. 12). – Mairhofer Nr. 49. (217ra) Mairhofer Nr. 50. (217ra) Mairhofer Nr. 52. (217rb) Mairhofer Nr. 33. (217va) Graesse Nr. 51 (50), 2. – Mairhofer Nr. 53. (217va) Graesse Nr. 51 (50), 3. – Mairhofer Nr. 54. (218ra) Mairhofer Nr. 55. (218rb) Mairhofer Nr. 56. (218rb) Graesse Nr. 119 (114), 7. – Mairhofer Nr. 57. (218va) Graesse Nr. 119 (114), 3. – Mairhofer Nr. 58. (219ra) Graesse Nr. 119 (114), 4. – Mairhofer Nr. 59. (219va) Graesse Nr. 131 (126), 9. – Mairhofer Nr. 60. (219vb) Mairhofer Nr. 61. (219vb) Graesse Nr. 119 (114), 5. – Mairhofer Nr. 62. (220ra) Graesse Nr. 131 (126), 4. – Mairhofer Nr. 63. (220rb) Graesse Nr. 131 (126), 5. – Mairhofer Nr. 64. (220rb) Mairhofer Nr. 65. (220vb) Graesse Nr. 131 (126), 6. – Mairhofer Nr. 66. (221ra) Mairhofer Nr. 67. (221rb) Mairhofer Nr. 68. (221va) Ed.: A. E. Schönbach, Stu dien zur Erzählungsliteratur des Mittelalters (SB Wien 144). Wien 1901, 57–59. – Mairhofer Nr. 69. 11.3 (222ra–229ra) Dist. 3: Miracula de Iohanne Baptista et sanctis apostolis atque martyribus. (222ra) Tit.: Incipit distinccio tercia in qua agitur de miraculis sancti Iohannis Baptiste et sanc torum apostolorum atque martirum etc. Sequuntur capitula. (222rb) Inhaltsübersicht. (222va–b) Iohannes Baptista: (222va) Graesse Nr. 86 (81), 3. – Mairhofer Nr. 70. (222va) Graesse 125 (120), 2. – Mairhofer Nr. 71. (222va) Graesse Nr. 125 (120), 4. – Mairhofer Nr. 72. (222vb–223vb) Iohannes ap. et ev.: (222vb) Graesse Nr. 9, 1. – Mairhofer Nr. 73. (222vb) Graesse Nr. 9, 3–4. – Mairhofer Nr. 74. (223rb) Graesse Nr. 9, 5. – Mairhofer Nr. 75. (223va) Graesse Nr. 9, 8. – Mairhofer Nr. 76. (223vb) Graesse Nr. 9, 11. – Mairhofer Nr. 77. (223vb–224rb) Petrus ap.: (223vb) Graesse Nr. 89 (84), 4 (S. 378, Z. 9–22). – Mairhofer Nr. 78. (224ra) Graesse Nr. 89 (84), 4 (S. 378, Z. 22–S. 379, Z. 6). – Mairhofer Nr. 79. (224ra) Graesse Nr. 89 (84), 4 (S. 379, Z. 6–14). – Mairhofer Nr. 80. (224rb) Mairhofer Nr. 81. (224rb–225rb) Andreas ap.: (224rb) Graesse Nr. 2, 9. – Mairhofer 82. (225ra) Graesse Nr. 2, 10. – Mairhofer Nr. 83. (225rb–226ra) Iacobus maior: (225rb) Graesse Nr. 99 (94), 4. – Mairhofer Nr. 84. (225rb) Graes se Nr. 99 (94), 5. – Mairhofer Nr. 85. (225va) Graesse Nr. 99 (94), 6. – Mairhofer Nr. 86. (225va) Graesse Nr. 99 (94), 8. – Mairhofer Nr. 87. (225vb) Graesse Nr. 99 (94), 9. – Mairhofer Nr. 88. (225vb) Graesse Nr. 99 (94), 10. – Mairhofer Nr. 89. (226ra) Thomas ap.: Mairhofer Nr. 90. (226va–227ra) Marcus ev.: (226va) Tit.: De beato Marco ewangelista et eius transsitu (!) ad Venetos (Graesse Nr. 59 [57], 2 [S. 267, Z. 28–S. 268, Z. 14]. – Mairhofer Nr. 91). (226vb) Graesse Nr. 59 (57), 9. – Mairhofer Nr. 92. 138 Cod. 125 (227ra–vb) Stephanus: (227ra) Graesse Nr. 8, 6. – Mairhofer Nr. 93. (227rb) Graesse Nr. 112 (107) (S. 463, Z. 30–S. 464, Z. 16). – Mairhofer Nr. 94. (227va) Graesse Nr. 112 (107) (S. 464, Z. 16–S. 465, Z. 8). – Mairhofer Nr. 95. (227vb–228ra) Laurentius: (227vb) Graesse Nr. 117 (112), 5. – Mairhofer Nr. 96. (227vb) Graesse Nr. 117 (112), 7 (S. 495, Z. 11–32). – Mairhofer Nr. 98. (228ra–b) Thomas Cantuariensis: (228ra) Tit.: Qualiter et ubi passus sit beatus Thomas Cantua riensis (Graesse Nr. 11, 2 [S. 68, Z. 19–30]. – Mairhofer Nr. 99). (228ra) Tit.: De sancto Thoma et cantu angelorum (Graesse Nr. 11, 3 [S. 68, Z. 30–S. 69, Z. 3]. – Mairhofer Nr. 100). (228rb) Graesse Nr. 11, 3 (S. 69, Z. 7–13). – Mairhofer Nr. 101. (228rb) Sebastianus: Graesse Nr. 23, 4. – Mairhofer Nr. 102. (228rb) Blasius: Tit.: De sancto Blasio et porco et lupo (Graesse Nr. 38 [S. 167, Z. 30–S. 168, Z. 16]. – Mairhofer Nr. 103). (228va) Marcellinus: Ed.: A. Sorbelli, Corpus chronicorum Bononiensium (Rerum Italicarum scriptores. Raccolta degli storici italiani dal cinquecento al millecinquecento 28/1). Città di Castello 1905, 206, Z. 18–207, Z. 33 (Cronaca A). – Mairhofer Nr. 104. (228vb) Mauritius: (228vb) Graesse Nr. 141 (136), 2. – Mairhofer Nr. 105. (228vb) Tit.: De sanc to Mauricio et clero ambicioso (Graesse Nr. 141 [136], 5. – Mairhofer Nr. 106). (228vb) Cosmas et Damianus: (228vb) Tit.: De sanctis Cosma et Damiano et viro et muliere (Graesse Nr. 143 [138], 3. – Mairhofer Nr. 107). 11.4 (229ra–238ra) Dist. 4: Miracula de episcopis. (229ra) Tit.: Incipit quarta distinccio de sanctis doctoribus Augustino Gregorio Ambrosio Martino et Nicolao episcopis etc. (229ra) Inhaltsübersicht. (229rb–230vb) Augustinus: (229rb) Graesse Nr. 124 (119), 4 (S. 562, Z. 15–20). – Mairhofer Nr. 108. (229va) Mairhofer Nr. 109. (229vb) Graesse Nr. 124 (119), 5. – Mairhofer Nr. 110. (229vb) Graesse Nr. 124 (119), 6. – Mairhofer Nr. 111. (230ra) Graesse Nr. 124 (119), 7. – Mairhofer Nr. 598. (230ra) Graesse Nr. 124 (119), 8. – Mairhofer Nr. 112. (230rb) Graesse Nr. 124 (119), 13 (S. 565, Z. 5–29). – Mairhofer Nr. 113. (230va) Graesse Nr. 124 (119), 9. – Mairhofer Nr. 599. (230va) Graesse Nr. 124 (119), 10. – Mairhofer Nr. 600. (230va) Graesse Nr. 124 (119), 11. – Mairhofer Nr. 601. (230vb–233rb) Gregorius: (230vb) Graesse Nr. 46, 2. – Mairhofer Nr. 114. (230vb) Tit.: De pue ris anglicanis (Graesse Nr. 46, 3 [S. 190, Z. 1–29]. – Mairhofer Nr. 115). (231ra) Graesse Nr. 46, 4 (S. 190, Z. 39–S. 191, Z. 1 und S. 191, Z. 37–S. 192, Z. 16). – Mairhofer Nr. 116. (231rb) Graes se Nr. 46, 6 (S. 194, Z. 7–20). – Mairhofer Nr. 117. (231rb) Graesse Nr. 46, 6 (S. 194, Z. 20– 39). – Mairhofer Nr. 118. (231va) Graesse Nr. 46, 11–12. – Mairhofer Nr. 119. (231va) Graesse Nr. 46, 13. – Mairhofer Nr. 120. (231vb) Graesse Nr. 46, 16. – Mairhofer Nr. 121. (232ra) Tit.: De monacho qui peculium restituit et mortuos revixit et fratres detrahentes increpavit(Graesse Nr. 46, 17. – Mairhofer Nr. 122). (232rb) Graesse Nr. 46, 1. – Mairhofer Nr. 602. (232va) Graesse Nr. 46, 4 (S. 191, Z. 10–18). – Mairhofer Nr. 603. (232va) Graesse Nr. 46, 5 (S. 193, Z. 38–S. 194, Z. 6). – Mairhofer Nr. 604. (232va) Graesse Nr. 46, 7. – Mairhofer Nr. 605. (232vb) Graesse Nr. 46, 8 (S. 195, Z. 30–S. 196, Z.1). – Mairhofer Nr. 606. (232vb) Graesse Nr. 46, 8 (S. 196, Z. 3–11). – Mairhofer Nr. 607. (233ra) Graesse Nr. 46, 14. – Mairhofer Nr. 608. (233ra) Graesse Nr. 46, 19. – Mairhofer Nr. 609. (233rb–234vb) Ambrosius: (233rb) Graesse Nr. 57 (55), 1 (S. 250, Z. 28–S. 251, Z.13). – Mairhofer Nr. 123. (233rb) Graesse Nr. 57 (55), 5. – Mairhofer Nr. 124. (233va) Graesse Nr. 57 (55), 2 (S. 251, Z. 21–32). – Mairhofer Nr. 610. (233vb) Graesse Nr. 57 (55), 2 (S. 251, Z. 32–S. 252, Z. 1). – Mairhofer Nr. 611. (233vb) Graesse Nr. 57 (55), 3 (S. 252, Z. 16–25). – Mairhofer Nr. 612. (233vb) Graesse Nr. 57 (55), 4 (S. 253, Z. 4–8). – Mairhofer Nr. 613. (233vb) Cod. 125 139 Graesse Nr. 57 (55), 4 (S. 253, Z. 14–30). – Mairhofer Nr. 614. (234ra) Graesse Nr. 57 (55), 6 (S. 255, Z. 15–21). – Mairhofer Nr. 615. (234ra) Graesse Nr. 57 (55), 7. – Mairhofer Nr. 616. (234vb–236ra) Martinus: (234vb) Graesse Nr. 166 (161) (S. 741, Z. 29–S. 742, Z. 33). – Mairhofer Nr. 125. (234vb) Graesse Nr. 166 (161) (S. 742, Z. 33–39). – Mairhofer Nr. 126. (235ra) Graesse Nr. 166 (161) (S. 743, Z. 35–S. 744, Z. 10). – Mairhofer Nr. 127. (235ra) Tit.: De humilitate sancti Martini (Graesse Nr. 166 [161] [S. 745, Z. 5–20]. – Mairhofer Nr. 128). (235rb) Graesse Nr. 166 (161) (S. 745, Z. 21–31). – Mairhofer Nr. 129. (235rb) Tit.: De largitate eius (Graesse Nr. 166 [161] [S. 746, Z. 29–S. 747, Z. 16]. – Mairhofer Nr. 130). (235va) Graesse Nr. 166 (161) (S. 749, Z. 2–14). – Mairhofer Nr. 131. (235vb) Graesse Nr. 166 (161) (S. 744, Z. 33–S. 745, Z. 5). – Mairhofer Nr. 617. (235vb) Tit.: De Severo et Martino (Graesse Nr. 166 [161] [S. 749, Z. 14–21]. – Mairhofer Nr. 618). (235vb) Tit.: De exsequiis eius et Ambrosio (Graesse Nr. 166 [161] [S. 749, Z. 22–34]. – Mairhofer Nr. 619). (236ra) Tit.: De duobus sociis ceco et contracto (Graesse Nr. 166 [161] [S. 750, Z. 7–20]. – Mairhofer Nr. 620). (236ra–238ra) Nicolaus: (236ra) Graesse Nr. 3, 4. – Mairhofer Nr. 132. (236rb) Graesse Nr. 3, 6 (S. 25, Z. 11–24). – Mairhofer Nr. 133. (236rb) Graesse Nr. 3, 6 (S. 25, Z. 6–11 und Z. 24–S. 26, Z. 26). – Mairhofer Nr. 134. (236vb) Graesse Nr. 3, 8. – Mairhofer Nr. 135. (236vb) Tit.: De ymagine sancti Nicolai plagata (Graesse Nr. 3, 9. – Mairhofer Nr. 136). (237ra) Tit.: De quodam fatuo sapienti qui sanctum Nicolaum honoravit (Mairhofer Nr. 137). (237va) Graesse Nr. 3, 10. – Mairhofer Nr. 621. (237va) Graesse Nr. 3, 11. – Mairhofer Nr. 622. (237vb) Mairhofer Nr. 623. 11.5 (238ra–259ra) Dist. 5: Miracula de confessoribus. (238ra) Tit.: Quinta distinccio de confessoribus. (238ra) Inhaltsübersicht. (238va–239ra) Remigius: (238va) Tit.: De miraculo sancti Remigii episcopi (Graesse Nr. 16 [S. 96, Z. 5–15]. – Mairhofer Nr. 138). (238vb) Graesse Nr. 16 (S. 96, Z. 35–S. 97, Z. 28). – Mairhofer Nr. 139. (239ra–240va) Basilius: (239ra) Tit.: De famulo qui se dyabolo dedit et Basilio (Graesse Nr. 26, 5. – Mairhofer Nr. 140). (239vb) Tit.: Basilius (Graesse Nr. 26, 1. – Mairhofer Nr. 625). (240ra) Graesse Nr. 26,6. – Mairhofer Nr. 626. (240rb) Graesse Nr. 26, 7. – Mairhofer Nr. 627. (240va–241vb) Iohannes elemosinarius: (240va) Graesse Nr. 27, 1 (S. 126, Z. 19–28). – Mairho fer Nr. 141. (240va) Graesse Nr. 27, 1 (S. 126, Z. 28–S. 127, Z. 1). – Mairhofer Nr. 142. (240va) Graesse Nr. 27, 3 (S. 127, Z. 1–24). – Mairhofer Nr. 143. (240vb) Tit.: De precioso operimento (Graesse Nr. 27, 8. – Mairhofer Nr. 145). (241ra) Tit.: De populo exeunti ecclesiam (Graesse Nr. 27, 11. – Mairhofer Nr. 146). (241ra) Tit. (nach Mairhofer zu Nr. 631): Quomodo homines ad pecuniam pervocabat (Graesse Nr. 27, 4. – Mairhofer Nr. 630). (241rb) Tit. (nach Mairhofer zu Nr. 630): De magna eius largitate (Graesse Nr. 27, 9. – Mairhofer Nr. 631). (241va) Graesse Nr. 27, 3. – Mairhofer Nr. 144. (241vb) Petronella: Graesse Nr. 88 (83) (S. 368, Z. 2–12). – Mairhofer Nr. 147. (242ra–va) Germanus: (242ra) Graesse Nr. 107 (102) (S. 448, Z. 27–S. 449, Z. 12). – Mairhofer Nr. 148. (242ra) Graesse Nr. 107 (102) (S. 449, Z. 22–30). – Mairhofer Nr. 149. (242rb) Graesse Nr. 107 (102) (S. 449, Z.30–S. 450, Z.11). – Mairhofer Nr. 150. (242rb) Graesse Nr. 107 (102) (S. 450, Z. 33–39). – Mairhofer Nr. 151. (242rb) Graesse Nr. 107 (102) (S. 451, Z. 7–16). – Mairhofer Nr. 633. (242rb) Graesse Nr. 107 (102) (S. 450, Z. 11–24). – Mairhofer Nr. 632. (242va–b) Iohannes Chrysostomus: (242va) Graesse Nr. 138 (131) (S. 616, Z. 25–29). – Mairho fer Nr. 152. (242va) Graesse Nr. 138 (131) (S. 616, Z. 30–34). – Mairhofer Nr. 153. (242va) Graesse Nr. 138 (131) (S. 616, Z. 34–S. 617, Z. 5). – Mairhofer 154. (242vb–243va) Patricius: (242vb) Graesse Nr. 50 (49) (S. 213, Z. 17–23). – Mairhofer Nr. 155. (242vb) Graesse Nr. 50 (49) (S. 213, Z. 29–S. 216, Z. 13). – Mairhofer Nr. 156. 140 Cod. 125 (243va–244ra) Paulus eremita: (243va) Graesse Nr. 15 (S. 94, Z. 9–28). – Mairhofer Nr. 157. (243vb) Graesse Nr. 15 (S. 94, Z. 28–S. 95, Z. 18). – Mairhofer Nr. 158. (244ra) Tit.: De transitu eiusdem Pauli (Graesse Nr. 15 [S. 95, Z. 18–27]. – Mairhofer Nr. 159). (244ra–245ra) Antonius: (244ra) Graesse Nr. 21, 1 (S. 104, Z. 8–12). – Mairhofer Nr. 160. (244rb) Graesse Nr. 21,1 (S. 104, Z. 12–17). – Mairhofer Nr. 161. (244rb) Tit.: Quod demones Antonium verberaverunt (Graesse Nr. 21, 1 [S. 104, Z. 17–32]. – Mairhofer Nr. 162). (244rb) Graesse Nr. 21, 5 (S. 106, Z. 19–23). – Mairhofer Nr. 637. (244rb) Graesse Nr. 21, 3 (S. 105, Z. 22–29). – Mairhofer Nr. 634. (244va) Graesse Nr. 21, 4 (S. 106, Z. 2–11). – Mairhofer Nr. 635. (244va) Graesse Nr. 21, 5 (S. 106, Z. 10–18). – Mairhofer Nr. 636. (244va) Graesse Nr. 21, 5 (S. 106, Z. 23–27. – Mairhofer Nr. 638. (244vb) Graesse Nr. 21, 5 (S. 106, Z. 27–36). – Mairho fer Nr. 639. (244vb) Tit.: De digitorum exustione notabile (Mairhofer Nr. 242). (245ra–b) Macarius: (245ra) Graesse Nr. 18 (S. 101, Z. 5–8). – Mairhofer Nr. 163. (245ra) Graes se Nr. 18 (S. 101, Z. 8–23). – Mairhofer Nr. 164. (245rb) Graesse Nr. 18 (S. 101, Z. 23–32). – Mairhofer Nr. 165. (245rb) Tit.: Item de sancto Machario (Graesse Nr. 18 [S. 100, Z. 16–25]. – Mairhofer Nr. 640). (245rb) Tit.: Quod humilitate dyabolum vicit (Graesse Nr. 18 [S. 100, Z. 25–S. 101, Z. 1]. – Mairhofer Nr. 641). (245rb–vb) Iohannes elemosinarius: (245rb) Graesse Nr. 27, 1 (S. 127, Z. 25–39). – Mairhofer Nr. 628. (245va) Graesse Nr. 27, 2 (S. 127, Z. 39–S. 128, Z. 28). – Mairhofer Nr. 629. (245vb–246ra) Pastor: (245vb) Graesse Nr. 175 (170) (S. 805, Z. 2–6). – Mairhofer Nr. 642. (245vb) Graesse Nr. 175 (170) (S. 805, Z. 6–11). – Mairhofer Nr. 643. (246ra) Moyses abb.: (246ra) Graesse Nr. 177 (172) (S. 806, Z. 22–26). – Mairhofer Nr. 644. (246ra) Graesse Nr. 177 (172) (S. 806, Z. 27–S. 807, Z. 2). – Mairhofer Nr. 645. (246ra) Graesse Nr. 177 (172) (S. 807, Z. 3–10). – Mairhofer Nr. 646. (246ra–b) Arsenius abb.: (246ra) Graesse Nr. 178 (163) (S. 808, Z. 30–37). – Mairhofer Nr. 647. (246ra) Graesse Nr. 178 (163) (S. 808, Z. 37–S. 809, Z. 7). – Mairhofer Nr. 648. (246rb) Graesse Nr. 178 (163) (S. 809, Z. 7–23). – Mairhofer Nr. 649. (246rb–va) Agathon abb.: (246rb) Graesse Nr. 179 (174) (S. 810, Z. 11–18). – Mairhofer Nr. 650. (246rb) Graesse Nr. 179 (174) (S. 810, Z. 18–26). – Mairhofer Nr. 651. (246va–248ra) Benedictus: (246va) Tit.: De gloriosis virtutibus Benedicti (Graesse Nr. 49 [48], 3 [S. 206, Z. 39–S. 207, Z. 24]. – Mairhofer Nr. 652). (246va) Graesse Nr. 49 (48), 4. – Mairho fer Nr. 653. (246vb) Graesse Nr. 49 (48), 7 (S. 208, Z. 37–S. 209, Z. 7). – Mairhofer Nr. 654. (246vb) Graesse Nr. 49 (48), 7 (S. 209, Z. 7–14). – Mairhofer Nr. 655. (246vb) ‹Ps.-Gregorius Magnus›: Dialogi, Lib. 2, cap. 12 (Ed.: F. Clark, The Pseudo-Gregorian dialogues 1. Leiden 1987. – Mairhofer Nr. 656). (247ra) Graesse Nr. 49 (48), 12. – Mairhofer Nr. 657. (247ra) Graesse Nr. 49 (48), 14. – Mairhofer Nr. 658. (247rb) Graesse Nr. 49 (48), 15 (S. 211, Z. 27– 36). – Mairhofer Nr. 659. (247rb) Tit.: De eiusdem virtutibus (Graesse Nr. 49 [48], 1 [S. 204, Z. 17–S. 205, Z. 1]. – Mairhofer Nr. 177). (247rb) Graesse Nr. 49 (48), 1 (S. 205, Z. 2–11). – Mairhofer Nr. 178. (247va) Graesse Nr. 49 (48), 2. – Mairhofer Nr. 179. (247va) Graesse Nr. 49 (48), 3 (S. 206, Z. 1–14). – Mairhofer Nr. 180. (247vb) Graesse Nr. 49 (48) (S. 206, Z. 29–39). – Mairhofer Nr. 181. (247vb) Graesse Nr. 49 (48) (S. 207, Z. 32–S. 208, Z. 7). – Mairhofer Nr. 182. (247vb) Graesse Nr. 49 (48), 6. – Mairhofer Nr. 183. (248ra) Graesse Nr. 49 (48), 16. – Mairho fer Nr. 184. (248ra–251vb) Bernardus: (248ra) Graesse Nr. 120 (115) (S. 529, Z. 13–29). – Mairhofer Nr. 663. (248rb) Graesse Nr. 120 (115) (S. 529, Z. 29–S. 530, Z. 9). – Mairhofer Nr. 185. (248va) Graesse Nr. 120 (115) (S. 530, Z. 25–S. 531, Z. 10). – Mairhofer Nr. 186. (248va) Graesse Nr. 120 (115) (S. 532, Z. 22–S. 533, Z. 2). – Mairhofer Nr. 187. (248vb) Graesse Nr. 120 (115) (S. 533, Z. 2–12). – Mairhofer Nr. 188. (248vb) Graesse Nr. 120 (115) (S. 534, Z. 30–S. 535, Z. 8). – Mairhofer Nr. 189. (249ra) Tit.: De demoniaco et Bernhardo (Graesse Nr. 120 [115] [S. 535, Z. 22–S. 536, Z. 10]. – Mairhofer Nr. 190). (249rb) Tit.: Item de demone peculanti et incubo (Graesse Nr. 120 [115] [S. 536, Z. 10–26]. – Mairhofer Nr. 191). (249va) Graesse Nr. 120 Cod. 125 141 (115) (S. 536, Z. 27–S. 537, Z. 2). – Mairhofer Nr. 192. (249va) Graesse Nr. 120 (115) (S. 537, Z. 3–12). – Mairhofer Nr. 193. (249va) Graesse Nr. 120 (115) (S. 537, Z. 13–24). – Mairhofer Nr. 194. (249vb) Graesse Nr. 120 (115) (S. 537, Z. 24–33). – Mairhofer Nr. 195. (249vb) Tit. (nach Mairhofer zu Nr. 197): Quod moriens fratribus tria observanda reliquit (Graesse Nr. 120 [115] [S. 537, Z. 33–S. 538, Z. 2]. – Mairhofer Nr. 196). (249vb) Tit. (nach Mairhofer zu Nr. 196): De poculo amaro militibus propinato (Graesse Nr. 120 [115] [S. 538, Z. 2–12]. – Mairhofer Nr. 197). (250ra) Graesse Nr. 120 (115) (S. 538, Z. 12–22). – Mairhofer Nr. 198. (250ra) Tit.: De virtutibus beati Bernhardi (Graesse Nr. 120 [115] [S. 528, Z. 16–22]. – Mairho fer Nr. 661). (250ra) Tit.: De obitu Bernhardi (Graesse Nr. 49 [48], 18. – Mairhofer Nr. 660). (250rb) Graesse Nr. 120 (115) (S. 534, Z. 15–30). – Mairhofer Nr. 662. (250rb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 15 (PL 185, 1324D–1325C. – Mairhofer Nr. 664). (250va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 16 (PL 185, 1325C–1326C. – Mairhofer Nr. 665). (250vb) Pastor: (250vb) Graesse Nr. 175 (170) (S. 803, Z. 15–26). – Mairhofer Nr. 166. (250vb) Graesse Nr. 175 (170) (S. 804, Z. 21–27). – Mairhofer Nr. 167. (250vb) Iohannes abb.: Graesse Nr. 176 (171) (S. 805, Z. 22–S. 806, Z. 6). – Mairhofer Nr. 168. (251ra) Moyses abb.: Graesse Nr. 177 (172) (S. 806, Z. 8–19). – Mairhofer Nr. 169. (251ra–va) Arsenius abb.: (251ra) Graesse Nr. 178 (163) (S. 807, Z. 14–26). – Mairhofer Nr. 170. (251rb) Graesse Nr. 178 (163) (S. 807, Z. 26–S. 808, Z. 2). – Mairhofer Nr. 171. (251rb) Graesse Nr. 178 (163) (S. 808, Z. 3–22). – Mairhofer Nr. 172. (251va) Agathon abb.: Graesse Nr. 179 (174) (S. 810, Z. 30–S. 811, Z. 3). – Mairhofer Nr. 173. (251va–252va) Hieronymus: (251va) Graesse Nr. 146 (141) (S. 654, Z. 2–25). – Mairhofer Nr. 174. (251vb) Graesse Nr. 146 (141) (S. 654, Z. 25–S. 655, Z. 19). – Mairhofer Nr. 175. (251vb) Graesse Nr. 146 (141) (S. 655, Z. 31–S. 657, Z. 13). – Mairhofer Nr. 176. (252va) Aegidius: Graesse Nr. 130 (125) (S. 584, Z. 2–14). – Mairhofer Nr. 199. (252va) Beda Venerabilis: Graesse Nr. 181 (176), 4 (S. 833, Z. 13–S. 834, Z. 2). – Mairhofer Nr. 200. (252vb) Udo Magdeburgensis: Ed.: A. E. Schönbach, Studien zur Erzählungsliteratur des Mittel alters (SB Wien 144). Wien 1901, 2–9. – Mairhofer Nr. 666. (254vb) Dionysius: Mairhofer Nr. 667. (255va–257ra) Dominicus: (255va) Graesse Nr. 113 (108) (S. 467, Z. 10–29). – Mairhofer Nr. 201. (255va) Graesse Nr. 113 (108) (S. 468, Z. 26–S. 469, Z. 11). – Mairhofer Nr. 202. (255vb) Graesse Nr. 113 (108) (S. 469, Z. 11–18). – Mairhofer Nr. 203. (255vb) Graesse Nr. 113 (108) (S. 470, Z. 3–28). – Mairhofer Nr. 204. (256ra) Graesse Nr. 113 (108) (S. 470, Z. 28–38). – Mairhofer Nr. 205. (256ra) Tit.: De converso vidente dyabolum (Graesse Nr. 113 [108] [S. 470, Z. 38–S. 471, Z. 13]. – Mairhofer Nr. 206). (256rb) Graesse Nr. 113 (108) (S. 471, Z. 13–19). – Mairhofer Nr. 207. (256rb) Graesse Nr. 113 (108) (S. 473, Z. 32–S. 474, Z. 5). – Mairhofer Nr. 209. (256va) Graesse Nr. 113 (108) (S. 474, Z. 5–10). – Mairhofer Nr. 210. (256va) Graesse Nr. 113 (108) (S. 476, Z. 25–31). – Mairhofer Nr. 211. (256va) Graesse Nr. 113 (108) (S. 477, Z. 20–S. 478, Z. 18). – Mairhofer Nr. 212. (256vb) Tit.: De morte et exhortacione eius (Graesse Nr. 113 [108] [S: 478, Z. 23–31]. – Mairhofer Nr. 213). (256vb) Graesse Nr. 113 (108) (S. 482, Z. 13–S. 483, Z. 8). – Mairhofer Nr. 214. (257ra–259ra) Franciscus: (257ra) Graesse Nr. 149 (144) (S. 663, Z. 6–22). – Mairhofer Nr. 215. (257rb) Tit.: Quod carnalis frater eum despexit (Graesse Nr. 149 [144] [S. 664, Z. 2–6]. – Mair hofer Nr. 216). (257rb) Graesse Nr. 149 (144) (S. 664, Z. 6–10 und Z. 16–20). – Mairhofer Nr. 217. (257rb) Graesse Nr. 149 (144) (S. 664, Z. 10–16). – Mairhofer Nr. 218. (257rb) Graesse Nr. 149 (144) (S. 664, Z. 20–29). – Mairhofer Nr. 219. (257va) Graesse Nr. 149 (144) (S. 664, Z. 29–35). – Mairhofer Nr. 220. (257va) Graesse Nr. 149 (144) (S. 665, Z. 15–25). – Mairhofer Nr. 221. (257va) Graesse Nr. 149 (144) (S. 665, Z. 8–15). – Mairhofer Nr. 222. (257vb) Graesse Nr. 149 (144) (S. 665, Z. 25–32). – Mairhofer Nr. 223. (257vb) Graesse Nr. 149 (144) (S. 666, 142 Cod. 125 Z. 33–S. 667, Z. 3). – Mairhofer Nr. 224. (257vb) Graesse Nr. 149 (144) (S. 666, Z. 15–33). – Mairhofer Nr. 225. (258ra) Graesse Nr. 149 (144) (S. 667, Z. 4–11). – Mairhofer Nr. 226. (258ra) Graesse Nr. 149 (144) (S. 667, Z. 11–20). – Mairhofer Nr. 227. (258ra) Graesse Nr. 149 (144) (S. 667, Z. 20–26). – Mairhofer Nr. 228. (258rb) Graesse Nr. 149 (144) (S. 668, Z. 18–30). – Mairhofer Nr. 229. (258rb) Graesse Nr. 149 (144) (S. 668, Z. 34–39). – Mairhofer Nr. 230. (258rb) Tit.: Quod fuit multe pietatis (Graesse Nr. 149 [144] [S. 668, Z. 39–S. 669, Z. 8]. – Mairhofer Nr. 231). (258rb) Tit.: Quod vituperari desiderabat (Graesse Nr. 149 [144] [S. 669, Z. 31–36]. – Mairhofer Nr. 232). (258va) Tit.: Quod avibus predicabat (Graesse Nr. 149 [144] [S. 670, Z. 19–27]. – Mairhofer Nr. 233). (258va) Tit.: Quod aves obedierunt ei (Graesse Nr. 149 [144] [S. 670, Z. 27–31]. – Mairhofer Nr. 234). (258va) Tit.: De eodem (Graesse Nr. 149 [144] [S. 670, Z. 7–14]. – Mairho fer Nr. 235). (258va) Graesse Nr. 149 (144) (S. 670, Z. 32–S. 671, Z. 1). – Mairhofer Nr. 236. (258vb) Graesse Nr. 149 (144) (S. 671, Z. 13–25). – Mairhofer Nr. 237. (258vb) Graesse Nr. 149 (144) (S. 671, Z. 29–35). – Mairhofer Nr. 238. (258vb) Graesse Nr. 149 (144) (S. 671, Z. 39–S. 672, Z. 8). – Mairhofer Nr. 239. (259ra) Graesse Nr. 149 (144) (S. 672, Z. 10–15). – Mairhofer Nr. 240. (259ra) Graesse Nr. 149 (144) (S. 673, Z. 6–15). – Mairhofer Nr. 241. 11.6 (259ra–268vb) Dist. 6: Miracula de virginibus et viduis. (259ra) Tit.: Sequitur distinccio sexta et est de virginibus et viduis. (259ra) Inhaltsübersicht. (259rb–261rb) Maria Magdalena: (259rb) Graesse Nr. 96 (90), 1 und 2 (S. 409, Z. 29–S. 413, Z. 8). – Mairhofer Nr. 250. (260ra) Graesse Nr. 96 (90), 2 (S. 413, Z. 10–21). – Mairhofer Nr. 251. (260rb) Tit.: De eius felici transitu ex hoc mundo (Graesse Nr. 96 [90], 2 [S. 413, Z. 21–S. 415, Z. 2]. – Mairhofer Nr. 252). (260va) Graesse Nr. 96 (90), 2 (S. 415, Z. 3–8). – Mairhofer Nr. 253. (260va) Tit.: De milite occiso et resuscitato (Graesse Nr. 96 [90], 4. – Mairho fer Nr. 254). (260vb) Graesse Nr. 96 (90), 8. – Mairhofer Nr. 256. (260vb) Graesse Nr. 96 (90), 7. – Mairhofer Nr. 255. (260vb) Graesse Nr. 96 (90), 9. – Mairhofer Nr. 257. (260vb) Graesse Nr. 96 (90), 5. – Mairhofer Nr. 258. (261ra) Graesse Nr. 96 (90), 10. – Mairhofer Nr. 259. (261ra) ‹Conradus Eberbacensis›: Exordium magnum Cisterciense, Dist. 4, cap. 2,1–30 (PL 185, 1096D–1097B. – Mairhofer Nr. 260). (261rb–263rb) Catharina: Tit.: De virtutibus sancte Katherine virginis et matris (Graesse Nr. 212 [209]. – Mairhofer Nr. 261). (262ra) Graesse Nr. 172 (167) (S. 795, Z. 5–9). – Mairhofer Nr. 262. (262ra) Graesse Nr. 172 (167) (S. 795, Z. 10–20). – Mairhofer Nr. 263. (262ra) Mairhofer Nr. 264. (262vb) Mairhofer Nr. 265. (263ra) Tit.: De sacerdote et cane cuius caput latrabat (Mairhofer Nr. 266). (263rb–va) Agnes: (263rb) Graesse Nr. 24,2 (S. 116, Z. 2–8). – Mairhofer Nr. 267. (263rb) Graes se Nr. 24, 3. – Mairhofer Nr. 268. (263va) Tit.: De ymagine sancte Agnetis (Graesse Nr. 24, 4 [S. 116, Z. 16–26]. – Mairhofer Nr. 269). (263va–b) Agatha: (263va) Graesse Nr. 39 (S. 173, Z. 4–14). – Mairhofer Nr. 270. (263va) Tit.: De velo sancte Agathe (Graesse Nr. 39 [S. 173, Z. 18–25]. – Mairhofer Nr. 271). (263vb–264ra) Undecim milia virginum: (263vb) Graesse Nr. 158 (153), 2. – Mairhofer Nr. 272. (263vb) Graesse Nr. 158 (153), 3. – Mairhofer Nr. 273. (264ra) Margarita: Graesse Nr. 151 (146). – Mairhofer Nr. 274. (264rb) Thais meretrix: Graesse Nr. 152 (147) (S. 677, Z. 25–S. 679, Z. 24). – Mairhofer Nr. 275. (264vb) Tit.: De alia meretrice (Graesse Nr. 152 [147] [S. 679, Z. 24–31]. – Mairhofer Nr. 276). (265ra) Eugenia, Prothus et Iacinthus: Graesse Nr. 136 (129) (S. 602, Z. 13–S. 604, Z. 31). – Mairhofer Nr. 277. (265va–268vb) Elisabeth: (265va) Tit.: De sancta Elizabeth lantgravia et viro eius (Mairhofer Nr. 278). (268rb) Tit.: De obitu Elizabeth et eius laude (Mairhofer Nr. 279). Cod. 125 143 11.7 (268vb–276va) Dist. 7: Miracula de defunctis. (268vb) Tit.: Sequitur distinccio septima et est ad diversas materias introductiva ut patet intuenti. (268vb) Inhaltsübersicht. (269ra) Omnes sancti: Graesse Nr. 162 (157) (S. 727, Z. 12–S. 728, Z. 4). – Mairhofer Nr. 280 (hier zweigeteilt: Tit. 1: De festo omnium sanctorum et eius iniciis. Tit. 2: De eodem). (269va–271rb) Commemoratio animarum: (269va) Graesse Nr. 163 (158) (S. 731, Z. 26–S. 732, Z. 9). – Mairhofer Nr. 282. (269va) Graesse Nr. 163 (158) (S. 731, Z. 4–24). – Mairhofer Nr. 281. (269vb) Graesse Nr. 163 (158) (S. 732, Z. 32–S. 733, Z. 11). – Mairhofer Nr. 284. (270ra) Graes se Nr. 163 (158) (S. 732, Z. 13–22). – Mairhofer Nr. 283. (270ra) Graesse Nr. 163 (158) (S. 733, Z. 20–25). – Mairhofer Nr. 285. (270ra) Graesse Nr. 163 (158) (S. 733, Z. 11–18). – Mairhofer Nr. 286. (270rb) Tit.: De fluvio infernali (Graesse Nr. 163 [158] [S. 733, Z. 30–S. 734, Z. 19]. – Mairhofer Nr. 287). (270rb) Mairhofer Nr. 288. (270va) Graesse Nr. 163 (158) (S. 734, Z. 33–S. 735, Z. 3). – Mairhofer Nr. 289. (270vb) Graesse Nr. 163 (158) (S. 735, Z. 18–26). – Mairhofer Nr. 290. (271ra) Graesse Nr. 163 (158) (S. 735, Z. 32–S. 736, Z. 8). – Mairhofer Nr. 291. (271ra) Graesse Nr. 163 (158) (S. 736, Z. 9–13). – Mairhofer Nr. 292. (271ra) Graesse Nr. 163 (158) (S. 738, Z. 34–S. 739, Z. 13). – Mairhofer Nr. 293. (271rb) Graesse Nr. 163 (158) (S. 739, Z. 19–29). – Mairhofer Nr. 294. (271rb) Ed.: J. Klapper, Exempla aus Handschriften des Mittelalters (Sammlung mittellatei nischer Texte II). Heidelberg 1911, Nr. 26, 27. – Mairhofer Nr. 295. (271va) Mairhofer Nr. 296. (271va) Mairhofer Nr. 297. (271va) Mairhofer Nr. 298. (271vb) Mairhofer Nr. 299. (271Ira) ‹Innocentius III.›: De contemptu mundi, Lib. 3, cap. 7 (PL 217, 740A. – Mairhofer Nr. 300). (271Ira) Mairhofer Nr. 301. (271Iva–272rb) ‹Honorius Augustodunensis›: Speculum ecclesiae. Sermo generalis: (271Iva) Ad milites (PL 172, 865A–C. – Mairhofer Nr. 302). (271Ivb) Ad pauperes (PL 172, 864C–865A. – Mairhofer Nr. 303). (271Ivb) Ad agricolas (PL 172, 866B–867B. – Mairhofer Nr. 304). (272rb) Ad coniugatos (PL 172, 870A–B. – Mairhofer Nr. 305). (272rb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 33 (PL 185, 1344B–D) = ‹Conradus Eberbacensis›: Exordium magnum Cisterciense, Dist. 4, cap. 35, 1–48 (PL 185, 1123B–1124A). – Mairhofer Nr. 306. (272va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 34 (PL 185, 1344D–1345B) = ‹Conradus Eberbacensis›: Exordium magnum Cister ciense, Dist. 5, cap. 2, 41–64 (PL 185, 1126D–1127B). – Mairhofer Nr. 307. (272va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 35 (PL 185, 1345C–D) = ‹Conradus Eberbacensis›: Exordium magnum Cisterciense, Dist. 5, cap. 3, 11–44 (PL 185, 1127C–1128B). – Mairhofer Nr. 308. (272vb–273va) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum: (272vb) Lib. 2, cap. 204, 1–6 (PL 179, 1188B–1189C. – Mairhofer Nr. 309). (273rb) Lib. 2, cap. 204, 7 (PL 179, 1189D–1190A. – Mairhofer Nr. 310). (273rb) Lib. 3, cap. 293, 1–2 (PL 179, 1262A–C. – Mairho fer Nr. 311). (273va) Mairhofer Nr. 312. (273vb) Mairhofer Nr. 313. (274rb) Mairhofer Nr. 314. (274va) Mairhofer Nr. 315. (275ra) Mairhofer Nr. 316. (275rb) Mairhofer Nr. 317 (Tit. auf Bl. 275va wiederholt). (275va–276rb) ‹Gregorius Magnus›: Dialogi, Lib. 4: (275va) Cap. 33 (PL 77, 372C–373A. – Mairhofer Nr. 318). (275vb) Cap. 40 (PL 77, 392C–393A. – Mairhofer Nr. 319). (275vb) Cap. 40 (PL 77, 393B–C. – Mairhofer Nr. 320). (276ra) Cap. 54 (PL 77, 413B–C. – Mairhofer Nr. 321). (276ra) Cap. 55 (PL 77, 416A–B. – Mairhofer Nr. 322). (276rb) Cap. 56 (PL 77, 416B–C. – Mairhofer Nr. 323). (276rb) Cap. 53 (PL 77, 412C–413 A. – Mairhofer Nr. 324). (276rb) Graesse Nr. 180 (175) (S. 816, Z. 12–33). – Mairhofer Nr. 325. 11.8 (276va–286ra) Dist. 8: Miracula de sacramentis ecclesiae. (276va) Inhaltsübersicht. 144 Cod. 125 (276vb–278rb) Sacramentum baptismi: (276vb) Graesse Nr. 52, 2. – Mairhofer Nr. 326. (277rb) Tit.: Quod baptismus confert sanitatem corporis (Graesse Nr. 117 [112] [S. 491, Z. 1–7]; an schließend Ergänzung zur hl. Odilia, vgl. Graesse Nr. 190 [189]. – Mairhofer Nr. 327). (277rb) Graesse Nr. 181 (176), 1 (S. 824, Z. 24–28). – Mairhofer Nr. 328. (277rb) Graesse Nr. 181 (176) (S. 834, Z. 3–10). – Mairhofer Nr. 329. (277va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 32 (PL 185, 1377B–1378C. – Mairhofer Nr. 330). (277vb) Mairhofer Nr. 331. (278rb) Sacramentum confirmationis: Mairhofer Nr. 332. (278rb–va) Unctio infirmorum: (278rb) Mairhofer Nr. 333. (278va) Mairhofer Nr. 334. (278vb–280ra) Sacramentum eucharistiae: (278vb–279rb) ‹Paschasius Radbertus›: De corpore et sanguine dni, cap. 14: (278vb) PL 120, 1317B–C. – Mairhofer Nr. 335. (278vb) PL 120, 1318B–1319C. – Mairhofer Nr. 336. (279ra) PL 120, 1319D–1320D. – Mairhofer Nr. 337. (279rb) Mairhofer Nr. 338. (279va) Mairhofer Nr. 339. (279va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 1, cap. 20 (PL 185, 1296D–1297D. – Mairhofer Nr. 340). (279vb) Mairhofer Nr. 341. (279vb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 28–29 (PL 185, 1373D–1374D. – Mairhofer Nr. 342). (280ra) Mairhofer Nr. 343. (280ra–284vb) Sacramentum paenitentiae et reconciliationis: (280ra) Mairhofer Nr. 344. (280rb) Mairhofer Nr. 345. (280rb) ‹Grimlaicus›: Regula solitariorum, cap. 36 (PL 103, 625D–626A. – Mairhofer Nr. 346). (280va) Mairhofer Nr. 347. (280va) Mairhofer Nr. 348. (280vb) Mairhofer Nr. 349. (281ra) Mairhofer Nr. 350. (281rb) Mairhofer Nr. 351. (281va) Mairhofer Nr. 352. (281vb) ‹Sigbertus Gemblacensis›: Chronica 823 (PL 160, 157C. – Mairhofer Nr. 353). (282ra) ‹Odo de Cheriton›, Parabolae Nr. 188 (Ed. Hervieux IV 339. – Mairhofer Nr. 354). (282ra) Mairhofer Nr. 355. (282rb) Mairhofer Nr. 356. (282va) Graesse Nr. 181 (176), 4 (S. 834, Z. 17– 21). – Mairhofer Nr. 357. (282va) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 175 (PL 179, 1146D–1147A. – Mairhofer Nr. 358). Tit. (nach Mairhofer zu Nr. 359): De quodam clerico vocali et eius constancia. (282vb) Tit. (nach Mairhofer zu Nr. 360): De clerico qui tonsuram deposuit (Mairhofer Nr. 359). (282vb) ‹Herbertus Claraevallensis›: De miracu lis, Lib. 3, cap. 13 (PL 185, 1365C–D. – Mairhofer Nr. 360). (283ra) Mairhofer Nr. 361. (283ra) Mairhofer Nr. 362. (283rb) Mairhofer Nr. 363. (283va) Mairhofer Nr. 364. (283va) ‹Sulpicius Severus›: Dialogi, Lib. 1, cap. 14 (PL 20, 192D–193B. – Mairhofer Nr. 365). (283vb) Tit.: De latrone qui per IIII candelas veniam meruit (Mairhofer Nr. 366). (284ra) Tit. (nach Mairhofer zu Nr. 368): De monacho quem confudit obsessus (Mairhofer Nr. 367). (284ra) Tit.: De confessione notandum (Mairhofer Nr. 368). (284ra) ‹Caesarius Heisterbacensis›: Dialogus miraculorum, Dist. 3, cap. 6 (Ed. Strange I, 116–120. – Mairhofer Nr. 369). Tit.: De virgine non confessa quam dyabolus confudit. (284rb) Mairhofer Nr. 370. (284va) Mairhofer Nr. 371. (284va) Mairhofer Nr. 372. (284va) Mairhofer Nr. 373. (284va) Mairhofer Nr. 374. (284vb) Mairhofer Nr. 375. (284vb–285va) Sacramentum matrimonii: (284vb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 34 (PL 185, 1379C–1381B. – Mairhofer Nr. 376). (285rb) Mairhofer Nr. 377. (285rb) Mairhofer Nr. 378. (285va) Mairhofer Nr. 379. 11.9 (286ra–311rb) Dist. 9: Miracula de factis et dictis theologorum. (286ra) Inhaltsübersicht. (287ra) Ed.: R. Lotti, Contributi su Domenico Cavalca (c. 1270–1342), Diss. Amsterdam 1987, 231–232. – Mairhofer Nr. 390. (287ra) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 174 (PL 179, 1146A–C. – Mairhofer Nr. 391). (287rb) Mairhofer Nr. 392. (287rb) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 207 (PL 179, 1192A–B. – Mairhofer Nr. 393). (287va) Mairhofer Nr. 394. (287vb) Mairhofer Nr. 395. (288rb–288Ira) ‹Sulpicius Severus›: Dialogi, Lib. 1: (288rb) Cap. 15 (PL 20, 193D–194B. – Mairhofer Nr. 397). (288rb) Cap. 17 (PL 20, 194D–195A. – Mairhofer Nr. 398). (288va) Cap. 18 (PL 20, 195C. – Mairhofer Nr. 399). Tit.: De incredibili obediencia novicii. (288va) Cap. 19 Cod. 125 145 (PL 20, 195D–196B. – Mairhofer Nr. 400). (288vb) Cap. 20 (PL 20, 196B–197A. – Mairhofer Nr. 401). (288Ira) ‹Wernerus de Ellerbach›: Deflorationes ss. patrum, Lib. 1, Sermo in rogationibus (PL 157, 968B–C. – Mairhofer Nr. 402). (288Ira–va) ‹Honorius Augustodunensis›: Speculum ecclesiae: (288Ira) Dnca 1. post pent. (PL 172, 1042B–D. – Mairhofer Nr. 403). (288Irb) Dnca 20. post pent. (PL 172, 1066D–1068A. – Mairhofer Nr. 404). (288Iva–289rb) ‹Gregorius Magnus›: XL Homiliae in Evangelia: (288Iva) Lib. 1 (PL 76, 1133C–1134B. – Mairhofer Nr. 405). (288Ivb) Lib. 2 (PL 76, 1300B–D. – Mairhofer Nr. 406). (289ra) Lib. 1 (PL 76, 1158B–1159A. – Mairhofer Nr. 407). (289rb–290vb) Ioseph: (289rb) Graesse Nr. 67 (63), 2 (S. 298, Z.15–S. 299, Z. 13). – Mairhofer Nr. 408. (289va) Graesse Nr. 67 (63), 2 (S. 299, Z. 15–S. 300, Z. 16). – Mairhofer Nr. 409. (289vb) Graesse Nr. 67 (63), 2 (S. 300, Z. 16–301, Z. 16). – Mairhofer Nr. 410. (290rb) Graesse Nr. 67 (63), 2 (S. 301, Z. 16–36). – Mairhofer Nr. 411. (290rb) Graesse Nr. 67 (63), 2 (S. 301, Z. 37–S. 302, Z. 11). – Mairhofer Nr. 412 (290va) Graesse Nr. 67 (63), 2 (S. 302, Z. 11–32). – Mairhofer Nr. 413 (290va) Graesse Nr. 67 (63), 2 (S. 302, Z. 32–S. 303, Z. 9). – Mairhofer Nr. 414. (290vb) Graesse Nr. 67 (63), 2 (S. 303, Z. 9–15). – Mairhofer Nr. 415 (290vb) Iudas Ischariot: Graesse Nr. 45 (S. 184, Z. 1–S. 186, Z. 13). – Mairhofer Nr. 416. (291va–292va) Pilatus: (291va) Graesse Nr. 53 (51) (S. 231, Z. 21–S. 232, Z. 24). – Mairhofer Nr. 417. (292ra) Graesse Nr. 53 (51) (S. 232, Z. 30–S. 233, Z. 26). – Mairhofer Nr. 418. (292rb) Graesse Nr. 53 (51) (S. 233, Z. 27–S. 234, Z. 26). – Mairhofer Nr. 419. (292va–293rb) Nero: (292va) Graesse Nr. 89 (84), 3 (S. 376, Z. 5–22). – Mairhofer Nr. 420. (292vb) Graesse Nr. 89 (84), 3 (S. 376, Z. 25–S. 377, Z. 14). – Mairhofer Nr. 421. (293ra) Graes se Nr. 89 (84), 3 (S. 377, Z. 15–29). – Mairhofer Nr. 422. (293rb–vb) Iulianus Apostata: (293rb) Graesse Nr. 30, 5 (S. 143, Z. 35–S. 144, Z. 11). – Mairho fer Nr. 423. (293rb) Graesse Nr. 30, 5 (S. 144, Z. 12–23). – Mairhofer Nr. 424. (293va) Graesse Nr. 125 (120), 1 (S. 571, Z. 3–8). – Mairhofer Nr. 425 (293va) Graesse Nr. 125 (120), 1 (S. 571, Z. 8–14). – Mairhofer Nr. 426. (293va) Graesse Nr. 125 (120), 1 (S. 571, Z. 14–21). – Mairhofer Nr. 427. (293vb) Graesse Nr. 125 (120), 1 (S. 571, Z. 21–28). – Mairhofer Nr. 428. (293vb) Graes se Nr. 30, 5 (S. 144, Z. 34–35). – Mairhofer Nr. 429. (293vb–294rb) Basilius: (293vb) Graesse Nr. 30, 5 (S. 144, Z. 35–S. 145, Z. 6). – Mairhofer Nr. 430. (294ra) Graesse Nr. 30, 5 (S. 145, Z. 6–30). – Mairhofer Nr. 431. (294rb) Graesse Nr. 181 (176), 4 (S. 838, Z. 9–15). – Mairhofer Nr. 432. (294rb) Graesse Nr. 181 (176), 4 (S. 837, Z. 31–38). – Mairhofer Nr. 433. (294rb) Graesse Nr. 181 (176), 4 (S. 837, Z. 13–21). – Mairhofer Nr. 434. (294va) Graesse Nr. 181 (176), 4 (S. 836, Z. 37–S. 837, Z. 10). – Mairhofer Nr. 435. (294va) Graesse Nr. 181 (176), 4 (S. 839, Z. 1–12). – Mairhofer Nr. 436. (294vb) Mairhofer Nr. 437. (295ra) Mairhofer Nr. 438. (295rb) Graesse Nr. 181 (176), 4 (S. 840, Z. 3–841, Z. 5). – Mairhofer Nr. 439. (295va) H 12553–12557. – Mairhofer Nr. 440. (295vb) Graesse Nr. 136 (129) (S. 603, Z. 4–9). – Mairhofer Nr. 441. (295vb) Mairhofer Nr. 442. (296ra) Graesse Nr. 182 (177) (S. 846, Z. 39–S. 847, Z. 14). – Mairhofer Nr. 443. (296ra) Graesse Nr. 182 (177) (S. 854, Z. 9–17). – Mairhofer Nr. 444. (296rb) Mairhofer Nr. 445. (296rb) Mairho ferNr. 446. (296va) 3. Esr 3,3–4,47. – Mairhofer Nr. 447. (297rb–300ra) ‹Odo de Cheriton›: Parabolae: (297rb) Nr. 6 (Ed. Hervieux IV 267. – Mairhofer Nr. 448). (297rb) Nr. 8 (Ed. Hervieux IV 268. – Mairhofer Nr. 449). (297va) Mairhofer Nr. 450. (297va) Nr. 12 und 13 (Ed. Hervieux IV 271. – Mairhofer Nr. 451). (297va) Nr. 26 (Ed. Hervieux IV 275. – Mairhofer Nr. 452). (297va) Nr. 42 (Ed. Hervieux IV 281. – Mairhofer Nr. 453). (297vb) Nr. 45 (Ed. Hervieux IV 282. – Mairhofer Nr. 454). (297vb) Nr. 46 (Ed. Hervieux IV 282. – Mairhofer Nr. 455). (298ra) Nr. 47 (Ed. Hervieux IV 283. – Mairhofer Nr. 456). (298ra) Nr. 52 (Ed. Hervieux IV 285. – Mairhofer Nr. 457). (298ra) Mairhofer Nr. 458. (298ra) Nr. 74 (Ed. Hervieux IV 294. – Mairhofer Nr. 459). (298rb) Nr. 85 (Ed. Hervieux IV 298. – Mairhofer 146 Cod. 125 Nr. 460). (298rb) Nr. 86 (Ed. Hervieux IV 298–299. – Mairhofer Nr. 461). (298rb) Nr. 88 (Ed. Hervieux IV 299. – Mairhofer Nr. 462). (298rb) Mairhofer Nr. 463. (298va) Nr. 93 (Ed. Hervieux IV 301. – Mairhofer Nr. 464). (298va) Mairhofer Nr. 465. (298va) Nr. 102 (Ed. Hervieux IV 304. – Mairhofer Nr. 466). (298vb) Nr. 105 (Ed. Hervieux IV 305. – Mairhofer Nr. 467). (298vb) Nr. 116 und 117 (Ed. Hervieux IV 309–310. – Mairhofer Nr. 468). (299ra) Vgl. Nr. 123 (Ed. Hervieux IV 312. – Mairhofer Nr. 469). (299ra) Nr. 132 (Ed. Hervieux IV 315–316. – Mairhofer Nr. 470). (299ra) Nr. 131 (Ed. Hervieux IV 315. – Mairhofer Nr. 471). (299rb) Nr. 136 (Ed. Hervieux IV 217. – Mairhofer Nr. 472). (299va) Nr. 133 (Ed. Hervieux IV 316. – Mairhofer Nr. 473). (299va) Nr. 143 (Ed. Hervieux IV 320–321. – Mairhofer Nr. 474). Tit.: De humilitate. (299va) Nr. 144 (Ed. Hervieux IV 321. – Mairhofer Nr. 475). (299vb) Nr. 153 (Ed. Hervieux IV 325. – Mairhofer Nr. 476). (299vb) Nr. 163 (Ed. Hervieux IV 328. – Nr. 477). (299vb) Nr. 164 (Ed. Hervieux IV 328–329. – Mairhofer Nr. 478). (299vb) Nr. 157 (Ed. Hervieux IV 329–330. – Mairhofer Nr. 479). (300ra) Nr. 171 (Ed. Hervieux IV 332. – Mairhofer Nr. 480). (300ra) Mairhofer Nr. 481. (300ra) Tit.: De libertate animi (Mairhofer Nr. 482). (300rb) Mairho fer Nr. 483. (300rb) Mairhofer Nr. 484. (300rb) Graesse Nr. 180 (175) (S. 815, Z. 24–S. 816, Z. 9. – Mairhofer Nr. 485). (300va) Mairhofer Nr. 486. (300va) Mairhofer Nr. 487. (300va–301ra) ‹Grimlaicus›: Regula solitariorum: (300va) Cap. 44 (PL 103, 635B–C. – Mairho fer Nr. 488). (300vb) Cap. 56 (PL 103, 647D–648A. – Mairhofer Nr. 489). (300vb) Cap. 54 (PL 103, 615D. – Mairhofer Nr. 490). (301ra) Cap. 57 (PL 103, 648D. – Mairhofer Nr. 491). (301ra) Cap. 39 (PL 103, 630B. – Mairhofer Nr. 492). (301ra) Mairhofer Nr. 493. (301rb) Mairhofer Nr. 494. (301rb) Vitae patrum, Lib. 5, Verba senio rum, libell. 16, cap. 19 (PL 73, 973B–C) = ‹Grimlaicus›: Regula solitariorum, cap. 59 (PL 103, 654A–B). – Mairhofer Nr. 495. (301va) Mairhofer Nr. 496. (301va) Mairhofer Nr. 497. (301va) Mairhofer Nr. 498. (301va) Mairhofer Nr. 499. (301vb) Mairhofer Nr. 500. (301vb) Mairhofer Nr. 501. (301vb) Mairhofer Nr. 502. (302ra) Mairhofer Nr. 503. (302ra) ‹Gregorius Magnus›: Dialogi, Lib. 4, cap. 11 (PL 77, 336B–337A. – Mairhofer Nr. 504). (302ra) Mairhofer Nr. 505. (302rb) Vitae patrum, Lib. 5, Verba seniorum, libell. 3, cap. 23 (PL 73, 864A–B. – Mairhofer Nr. 506). (302rb) Mairhofer Nr. 507. (302rb) Mairhofer Nr. 508. (302rb) Mairhofer Nr. 509. (302va) Mairhofer Nr. 510. (302va) Mairhofer Nr. 511. (302va) Mairhofer Nr. 512. (302vb) Vitae patrum, Lib. 5, Verba seniorum, libell. 6, cap. 18 (PL 73, 891C–D. – Mairhofer Nr. 513). (302vb) Mairhofer Nr. 514. (302vb) Mairhofer Nr. 515. (303ra) Vitae patrum, Lib. 5, Verba seniorum, libell. 6, cap. 21 (PL 73, 892B–C. – Mairhofer Nr. 516). (303ra) Mairhofer Nr. 517. (303ra) Mairhofer Nr. 518. (303rb) Vitae patrum, Lib. 5, Verba seniorum, libell. 3, cap. 14 (PL 73, 862B– C. – Mairhofer Nr. 519). (303rb) Mairhofer Nr. 520. (303va) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 55 (PL 73, 768C–D) = ‹Grimlaicus›: Regula solitariorum, cap. 39 (PL 103, 630C–D). – Mairhofer Nr. 521. (303va) ‹Hieronymus›: Epistola 107 (ad Lae tam) (PL 22, 872. – Mairhofer Nr. 522). (303va) Mairhofer Nr. 523. (303vb) Mairhofer Nr. 524. (303vb) Mairhofer Nr. 525. (303vb) Mairhofer Nr. 526. (304ra) Mairhofer Nr. 527. (304ra) Mairhofer Nr. 528. (304ra) Mairhofer Nr. 529. (304rb) Vgl. Vitae patrum, Lib. 5, Verba senio rum, libell. 8, cap. 24 (PL 73, 909C. – Mairhofer Nr. 530). (304rb) Mairhofer Nr. 531. (304rb) Mairhofer Nr. 532. (304va) Mairhofer Nr. 533. (304va) ‹Petrus Cantor›: Verbum abbreviatum, cap. 86 (PL 205, 256D–257A. – Mairhofer Nr. 534). (304va) ‹Nicolaus Pergamenus›: Dialogus creaturarum, dial. 75, 23–36 (Ed.: Th. Graesse, Die beiden ältesten lateinischen Fabelbücher des Mittelalters des Bischofs Cyrillus „Speculum sapientiae“ und des Nicolaus Pergamenus „Dialogus creaturarum“. Stuttgart 1880, 219. – Mairhofer Nr. 535). (304va) Mairhofer Nr. 536. (304vb) Mairhofer Nr. 537. (304vb) Mairhofer Nr. 538. (305ra) Mairhofer Nr. 539. (305ra) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 118 (PL 73, 782C–D. – Mairhofer Nr. 540). (305ra) Mairhofer Nr. 541. (305rb) Mairhofer Nr. 542. Tit.: Qui propriam iracundiam vindicant. (305rb) ‹Gregorius Magnus›: Dialogi, Lib. 4, cap. 19 (PL 77, 352A–B. – Mairhofer Nr. 543). (305rb) Mairhofer Nr. 544. (305va) Mairhofer Nr. 545. (305va) ‹Rufinus Aquileiensis›: Cod. 125 147 Historia monachorum, cap. 1 (PL 21, 394C. – Mairhofer Nr. 546). (305va) Mairhofer Nr. 547. (305va) Mairhofer Nr. 548. (305vb) Mairhofer Nr. 549. (305vb) Mairhofer Nr. 550. (305vb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 97 (PL 73, 778A–C) = Vitae patrum, Lib. 6, Verba seniorum, libell. 3, cap. 17 (PL 73, 1013D–1014B). – Mairhofer Nr. 551. (305vb) Mairhofer Nr. 552. (306ra) ‹Innocentius III.›: De contemptu mundi, Lib. 2: (306ra) Cap. 36 (PL 217, 731D–732A. – Mairhofer Nr. 553). (306ra) Cap. 39 (PL 217, 733B. – Mairhofer Nr. 554). (306ra) Cap. 20 (PL 217, 725A. – Mairhofer Nr. 555). (306ra) Mairhofer Nr. 556. (306rb–vb) Vitae patrum, Lib. 5: (306rb) Libell. 14, cap. 52 (PL 73, 963B–C. – Mairhofer Nr. 557). (306rb) Mairhofer Nr. 558. (306rb) Libell. 14, cap. 72 (PL 73, 966A–B. – Mairhofer Nr. 559). (306va) Libell. 5, cap. 34 (PL 73, 882D–883A. – Mairhofer Nr. 560). (306vb) Mairho fer Nr. 561. (306vb) Libell. 3, cap. 2 (PL 73, 860C–D. – Mairhofer Nr. 562). (306vb) Libell. 17, cap. 18 (PL 73, 976B. – Mairhofer Nr. 563). (306vb) Mairhofer Nr. 564. Tit.: De caritate. (306vb) Mairhofer Nr. 565. (307ra) ‹Petrus Cantor›: Verbum abbreviatum, cap. 98 (PL 205, 280A–B. – Mairhofer Nr. 566). Tit.: Quod caritas non agit properam. (307ra) Mairhofer Nr. 567. (307ra) ‹Gregorius Magnus›: Dialogi, Lib. 1, cap. 9 (PL 77, 197B–C. – Mairhofer Nr. 568). (307rb) Mairhofer Nr. 569. (307rb) ‹Rufinus Aquileiensis›: Historia monachorum, cap. 31 (PL 21, 458B–C. – Mairhofer Nr. 570). (307rb) Mairhofer Nr. 571. (307rb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 28 (PL 73, 756 B–C) und Vitae patrum, Lib. 6, Verba seniorum, libell. 2, cap. 17 (PL 73, 1004 A. – Mairhofer Nr. 572). (307va) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 13 (PL 73, 745B–746B. – Mairhofer Nr. 573). (307va) Mairhofer Nr. 574. Tit.: De obsidione castri. (307va) Mairhofer Nr. 575. (307vb) Mairhofer Nr. 576. (307vb) Mairhofer Nr. 577. (307vb) Mairhofer Nr. 578. (308ra) Mairhofer Nr. 579. (308va) Mairhofer Nr. 580. (308vb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 84 (PL 73, 775B– C. – Mairhofer Nr. 581). (309ra) ‹Sulpicius Severus, Iohannes Cassianus›: Vitae patrum, Lib. 4, Excerpta , cap. 43 (PL 73, 841A–C. – Mairhofer Nr. 582). (309ra) ‹Rufinus Aquileiensis›: Historia monachorum, cap. 1 (PL 21, 399A–400A. – Mairhofer Nr. 583). (309va) ‹Rufinus Aquileiensis›: Historia monachorum, cap. 6 (PL 21, 401C–403C. – Mairhofer Nr. 584). (309vb) ‹Sulpicius Severus, Iohannes Cassianus›: Vitae patrum, Lib. 4, Excerpta, cap. 52 (PL 73, 846B–D. – Mair hofer Nr. 585). (310ra) Vitae patrum, Lib. 5: (310ra) Libell. 4, cap. 27 (PL 73, 880C–D. – Mairhofer Nr. 586). (310ra) Libell. 5, cap. 37 (PL 73, 883D–884B. – Mairhofer Nr. 587). (310rb) Libell. 5, cap. 30 (PL 73, 881C. – Mairhofer Nr. 588). (310va) Libell. 7, cap. 41 (PL 73, 903A. – Mairhofer Nr. 589). (310va) Libell. 7, cap. 43 (PL 73, 903B–904A. – Mairhofer Nr. 590). (310vb) Libell. 10, cap. 10 (PL 73, 913C–D. – Mairhofer Nr. 591). (310vb) Libell. 14, cap. 18 (PL 73, 952A) = ‹Grimlaicus›: Regula solitariorum, cap. 58 (PL 103, 649D–650. – Mairhofer Nr. 592). (311ra) Vitae patrum, Lib 6: (311ra) Libell. 1, cap. 12 (PL 73, 994C–995A. – Mairhofer Nr. 593). (311ra) Libell. 1, cap. 13 (PL 73, 995A–B. – Mairhofer Nr. 594). Tit.: De fine boni hominis et mali hominis. (311rb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 54 (PL 73, 768B– C. – Mairhofer Nr. 595). (311rb) ‹Godefridus Altissiodorensis›: Declamationes ex s. Bernardi sermonibus, cap. 25 (PL 184, 454C–455A. – Mairhofer Nr. 597). (311rb) Schlussschrift: Et sic est finis. 148 12 Cod. 142 (311v) Miraculum de duobus sociis. Inc.: Erant duo socii simul coniuncti unus quesivit alterum quis eorum remocius ambulasset. Dixit primus Fui in provincia … – Expl.: … et ea que congregasti cuius erunt etc. (Ir–v) leer. P. A. / A. P. Cod. 142 REGULA S. AUGUSTINI. HUGO DE SANCTO VICTORE (?). LIBER ANNIVERSARIORUM NEOCELLENSIS. NECROLOGIUM NEOCELLENSE Perg. I, 173, I Bl. 300 × 230. Neustift, T. I: 1521, T. II: 1459. B: Zwei Teile: I (I, 1–51) und II (52–173, I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol (hier berücksichtigt). E: Renaissanceeinband: helles Leder über Holz. Neustift, 16. Jh. Leder insbes. am HD stark abgerieben, in den Ecken Holz tw. freiliegend. VD und HD gleich: vier von drei fachen Streicheisenlinien gebildete Rahmen (von außen nach innen): Rahmen 1: Dekor aus spitz aufeinander zulaufenden dreifachen Streicheisenlinien; Rahmen 2: Ranke mit eingeschlossenen Blüten und Granatapfel (Nr. 1); Rahmen 3: Ranke mit eingeschlossenen Blüten und Granatapfel (Nr. 2); Rahmen 4: Rundbogen auf Stab (Nr. 3). Mittelfeld oben und unten horizontal durch dreifache Streicheisenlinien begrenzt, in der Mitte Einzelstempel zwölfblättrige Rosette mit zwölfblättriger Mittelblüte frei (Nr. 4), in den Ecken Einzelstempel mit floralem Dekor frei (Nr. 5). Zwei Kurzriemen-Leder-Metall-Schließen. Kanten gerade, nach innen abgeschrägt und von zweifachen Streicheisenlinien eingefasst. Rücken: blau gefärbt (wie öfters in Neustift), vier Doppelbünde. Bünde durch dreifache Streicheisenlinien begrenzt, Linien am VD und HD fortgesetzt, spitz aufeinander zulaufend. Im zweiten Feld Signaturschild (s. bei G), im fünften Spuren eines abgelösten Papierschilds, leer. Kapitale mit naturfarbenem Spagat umstochen, oben nur mehr tw. erhalten. Spiegel Pap. 18. Jh. (?). Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. Unter dem Spiegel des VD Pergamentblatt, Gegenfalz zw. Bl. I und 1. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Auflösung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 G 4 (Bleistift) und Nr. 142 (2) (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Bl. 1r Stempel der ULB Tirol 19. Jh. Am Rücken gelbes Signaturschild der ULB Tirol 142/2. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. L: Wilhelm I 136. – Wretschko–Sprung 16. – O. Redlich, Tirolische Geschichtsquellen des Mittelalters (Fest schrift des Akademischen Historikerklubs in Innsbruck zur Erinnerung an das dreißigste Stiftungsfest). Innsbruck 1903, 8. – Sparber 226–227. – L. Santifaller, Calendarium Wintheri. Il più antico calendario ne crologio ed urbario del capitolo della cattedrale di Bressanone. Gleno 1926, 376–391, 413. – L. Santifaller, Kalendar und Nekrolog des Kollegiatstiftes im Kreuzgang zu Bressanone aus dem 13. Jahrhundert (Beihefte zum Jahrbuch für Geschichte, Kultur und Kunst 4). Bozen 1939, 11. – MGH Necrologia Germaniae III 28–47. – J. Baur, Die Spendung der Taufe in der Brixner Diözese in der Zeit vor dem Tridentinum (Schlern-Schriften 42). Innsbruck 1938, 142. – Goy 471, Nr. 85. – Neuhauser, Neustift 81. – Th. (H.) Innerhofer, Der Grabplatz Oswalds von Wolkenstein in der Stiftskirche von Neustift. Der Schlern 58 (1984) 418–419. – Katalog ULB Tirol II 113, 204. – Katalog ULB Tirol II, Beiheft, Taf. 66 und 79. – Kristeller VI 122. – Stefani 99–100. – Peintner, Stiftsbibliothek 122. I (I, 1–51) B: Perg. stark abgegriffen. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.?) + 6.IV48 + (II–1)51. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Gegenbl. zu Bl. 50 herausgeschnitten (Textverlust). Lage 1 Reklamante erhalten, beschnitten. In der Mitte des oberen Blattrandes rote Foliierung, dat. 1521 (s. bei A), in arabischen Ziffern 1–52. Cod. 142 149 S: Schriftraum 225 × 170, Bl. 1r–6v einspaltig, Bl. 7r–51v zwei Spalten, seitlich von roten Tintenlinien begrenzt, zu 23 Zeilen. Textualis formata (Textura), dat. 1521, Rubrizierung von gleicher Hand (dat. in Rot Bl. 4r, 6v, 22va, 29rb, 43vb; Bl. 4r zusätzlich in Schwarz). Schreiber: H. R. (?) (s. bei A). Bl. 45ra, Z. 11–23 von anderer Hand. Von späterer Hand I-Striche, bisweilen in Häkchenform, und U-Striche in gebogter Form. Vereinzelt interlineare Nachträge von zeitgenössischer Hand (in Tinte). A: Rote Auszeichnungsstriche, Trennstriche, Überschriften, Blattzählung. In den untersten Zeilen Unterlänge des Minuskelbuchstabens g bisweilen kalligraphisch gelängt. Bl. 5r Hinweiszeichen in Pfeilform (in Blei stift) von späterer Hand. Einzeilige rote Lombarden (pastos-erhabener Tintenauftrag, Bl. 30vb blaue Lom barde) mit einfachem Dekor: Punktverdickungen, Achterschlingen, Bl. 22va als Besatzmotiv mit schwarzer Tinte gezeichneter Profilkopf (mit geöffnetem Mund). Repräsentanten. Zwei größere Initialen (Bl. 1r: sechszeilig, rot-blau gespalten; Bl. 7ra: fünfzeilig, blau) mit rotem Fleuron née (charakteristische länglich-blasige Knospen) von der im 1. Fünftel 16. Jh. nachweisbaren Hand, die auch in Cod. 405 (dat. 1501, hier das Fleuronnée flüchtiger) und Cod. 194 (dat. 1507) tätig war, in beiden Bänden des Zollner-Graduales einige Initialen mit Fleuronnée (s. jeweils Ergänzungshand 2) geschmückt und Sign. 14820 zur Gänze ausgestattet hat. Bl. 25vb und Bl. 37va signiert H. R. (jeweils rot, in g-Unterlänge, letzte Zeile) und fügt Bl. 37va liniavit 1521 (in Rot) hinzu (lat. „liniare“ bzw. „lineare“: „Linien ausführen, mit Linien umgeben“, s. Du Cange et al., Glossarium mediae et infimae latinitatis. Niort 1883–1887, Bd. 5, Sp. 118a bzw. 115b; „lineare“ auch „eine Handschrift ausmalen“, s. E. Habel, F. Gröbel, Mittellateinisches Glossar. Paderborn 2008, Sp. 223); von dieser Hand auch die Datierung 1521 Bl. 4r, 6v, 22va, 29rb, 43vb.; unklar, ob H. R. lediglich die Be grenzungslinien des Schriftspiegels zog (allerdings fraglich, ob diese Tätigkeit eine Signatur rechtfertigte), oder als Rubrikator (und zugleich Schreiber) anzunehmen ist; es ist eher unwahrscheinlich, dass H. R. als Florator tätig war, denn dann wäre ihm die oben genannte Handschriftengruppe zuzuschreiben, was die Frage aufwirft, warum er ein Werk mit zwei Fleuronnée-Initialen signierte, ein vollständig fleuronniertes Buch wie Sign. 14820 aber nicht. S. R. 1 2 (Ir–v) leer. (1r–6v) Regula s. Augustini (PL 32, 1377–1384. Regula recepta = Ordo monasterii, Satz 1 + Praeceptum [CPL Nr. 1839b]. Vgl. L. Verheijen, La règle de saint Augustin 1 [Études augusti niennes 15,1]. Paris 1967, 417–437). (1r) Tit.: Regula beati Augustini. Daneben in Rot: 1521. (7r–51v) Hugo ‹de Sancto Victore (?)›: Expositio in regulam beati Augustini, unvollständig: Schluss fehlt (PL 176, 881–924C. – Goy 457–478). Tit.: Incipit glosa Hugonis super regulam beati Augustini. – Expl. mut: … Graciarum actio patri luminum semper [ II (52–173, I) B: Perg. stark abgegriffen, löchrig, Nahtspuren, insbes. am Blattrand stark verschmutzt. Bl. 53, 55, 65 und 67 jeweils mit angeklebtem Pergamentstreifen ergänzt. Buchblock nachträglich beschnitten (Text am oberen und seitlichen Blattrand bisweilen beschnitten). Lagen: (VII–2)63 + (VII–3)74 + (VII–2)86 + (VIII–3)97 + (VII–2)109 + (VII–2)121 + II125 + 5.IV166 + (III+1)173 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.?). Gegenbl. zu Bl. 54, 63, 64, 68, 74, 79, 85, 91, 97, 102, 108, 114 herausgeschnitten (ohne Textverlust). Gegenbl. zu Bl. 87 her ausgeschnitten (Textverlust, vgl. Schriftrest auf Falz zw. Bl. 97 und 98). Bl. 85 von kleinerem Format (295 × 190). Bl. 167 eingehängtes Einzelbl. Bl. 173 durch Pergamentmakulaturfalz verstärkt. Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Am oberen Blattrand ältere Foliierung in arabischen Ziffern 1–43 (= 53–121), nur die Blätter mit Nekrologeintragungen foliiert, die dazwischen befindlichen Blätter mit dem Text des Martyrologs unfoliiert. Bl. 52r Rest eines Signakels. S: Bl. 52r–126v Schriftraum ca. 220 × 160. Zwei Spalten, von roten bzw. rot-schwarzen bzw. schwarzen Tintenlinien gerahmt, Bl. 81r, 85r–v, 93r, 98r, 110r, 122r–126v einspaltig, zu meist 27 Zeilen. Bastarda 15. Jh. mit Nachträgen von späteren Händen. 150 Cod. 145 Bl. 127r–173r Schriftraum 240 × 180, von roten Tintenlinien in vier Felder unterteilt, jeweils ca. 120 × 80 (jedes Feld für einen Tag des Nekrologs). Haupthand Bastarda, dat. 1459 (Bl. 172v). Zahlreiche Nachtrags hände bis 2. H. 17. Jh. Schriftraum und Zeilenzahl mit Händen wechselnd. A: Rote Auszeichnungsstriche, Trennstriche, Paragraphzeichen, Durchstreichungen, Zeilenfüllsel, Über- und Schlussschriften. Bl. 122r Laus Deo mit roter Rahmung, das L mit Ausläufer, der in Herzblättern endet. KLLigaturen in Rot, Bl. 53r mit kordelartiger Aussparung, Bl. 99r mit kritzeligem schwarzem Fleuronnée. Rote Benediktionskreuze. Ein- und zweizeilige rote Lombarden, vereinzelt mit Punktverdickungen, cadellenarti gen Verzierungen und einfachem Fleuronnée in Schwarz; ab Bl. 123v lange schwarze I-Versalien, fallweise mit blassrotem Dekor. Repräsentanten. 3 (52ra–126v) Liber anniversariorum neocellensis (MGH Necrologia Germaniae III 39–47). Heiligenfeste aufgelistet in Santifaller, Calendarium 377–391 (Cal. n. 10). Folgende Blätter und Schriftspalten sind leer: Bl. 52ra–b, 58va–59rb, 64r, 70rb, 74rb, 75rb, 77rb, 78rb, 79rb, 80rb, 83rb, 84rb, 87rb, 88rb, 97rb, 101rb, 102rb, 103rb, 108rb, 120rb. 4 (127ra–172va) Necrologium Neocellense (MGH Necrologia Germaniae III 28–38). (172va) Datierung (rot): Explicit memoriale fratrum et sororum nostre confraternitatis finitum anno domini 1459 in vigilia concepcionis Marie virginis (7. Dezember 1459). (172vb) leer. (173ra) Nachtrag. Tit.: Utriusque sexus nomina laicalem nobiscum fraternitatem habentium hic infra notata sunt ab anno virginei partus 1506. (173rb) Datierter Nachtrag 1590: Ludovicus Ox scriba noster in fraternitatem nostram laicalem assumptus est. Anno domini 1590. Die 2 decembris anno 85 (? radiert). Obiit autem 7 die eiusdem anni et mensis. (173rb, unten) kurze Additionsrechnung. (173v) leer. (Ir) Bleistiftnotiz 20. Jh. (Datierung?). (Iv) leer. U. S. Cod. 145 BARTHOLOMAEUS A SANCTO CONCORDIO Pap. I, 314, I* Bl. 300 × 215. Süddeutscher Raum (Tirol?), 1427. B: Pap. bes. am oberen Blattrand gewellt, am seitlichen Rand bisweilen eingerissen und tw. fleckig, Bl. 1 Wasserschaden. Nachweise und Bilder der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Identische Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 663, T. II, Cod. 666, T. II, Cod. 787, T. II. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 11 + 26.VI313 + 1314 + (I–1)I* (Nachsatzbl. 18. Jh.). Ge genbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. 1 und 314 Einzelbl., wohl urspr. Vor- bzw. Nachsatzbl. Vor Bl. 1 und in den Lagenmitten von verschiedenen Händen beidseitig beschriebene und rubrizierte Pergamentfälze, längs geschnitten (vereinzelt Namen bzw. Namensanfänge erkennbar: möglicherweise aus einem Nekrolog 14. Jh.). Lagenreklamanten, tw. gerahmt, Bl. 301v weg geschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 205/210 × 145/150. Zwei Spalten, von Tintenlinien gerahmt, zu 37–46 Zeilen. Zirkelstiche. Bastarda von einer Hand mit unterschiedlichem Duktus, dat. 1427 (Bl. 313vb). Schreiber: Nicolaus Petrus. Bl. 1r und 314r–v Einträge von zeitgenössischer Hand. A: Rote Auszeichnungsstriche und -punkte, Paragraphzeichen (auch am seitlichen Blattrand), Rahmungen, Zeilenfüllsel, Unterstreichungen sowie Überschriften. Zwei- und drei-, vereinzelt auch vierzeilige rote Lombarden, tw. mit Punktverdickungen, vereinzelt mit einfachen Ausläufern. Bl. 120va I-Versal mit ein geschriebener (weiblicher?) Figur in Schwarz. Cod. 145 151 E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier Doppelbünde, von Gold bordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck BARTHOLOMAEI A S. CON CORDIO OPERA in Goldrahmung, im dritten Signaturschild (s. bei G). Kapitale mit Leder umschlagen. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`3 […] 6 […] (? radiert, Tinte), II 2 G 7 (Blei stift) und Nr. 145 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 145. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kaeppeli Nr. 436. – Katalog ULB Tirol II 117, 204–205. – Stefani 101. – Peintner, Stiftsbibliothek 121. 1 (Ir–v) leer. (1r) Versus de titulis librorum Decretalium (Druck: GW M23131. Vgl. Walther, Initia Nr. 18797). Die genaue Formulierung des jeweiligen Titulus ist über die Verse geschrieben. Inc.: Summum constitue rescriptum post aliud consue … – Expl. mut.: … Sponsans inpubes clam destina (!) sponsa duorum Condicio cleri quam polluit et lepro servus [ Forts. Nr. 3, Bl. 314r. Parallelüberlieferung: u. a. Innsbruck, ULB Tirol, Cod. 285, Bl. 57r; Freiburg, UB, Hs. 1283, Bl. 18v. (1v) leer bis auf Notiz: luo vinarii (?). 2 (2ra–313vb) Bartholomaeus a Sancto Concordio: Summa de casibus conscientiae (= Summa Pisanella) (Druck: GW 3450–3457. – Kaeppeli Nr. 436. Bloomfield Nr. 5052). (2ra) Abkürzungsverzeichnis. Tit.: In Christi nomine Amen. Iste sunt declaraciones de breviatu ris positis in hac summa. (2rb) Alphabetisches Stichwortverzeichnis. (6ra) Text (tw. vom Druck leicht abweichend). (101rb) Expl. mut. (Excomunicacio sexto): … seu excomunicatis recipere [ Forts. Bl. 102va– 103rb. Anschließend Bl. 101va Inc. mut.: ] volentes libro sexto salvis que dicuntur in sequenti … Bl. 102r Vermerk: ferte (!) duo follia (!) circa signum hoc = Verweis auf Forts. Bl. 103va. (311ra) Fünf Zeilen gegenüber Druck eingefügt, durchgestrichen … sola fama non sufficit ut (hier Einschub: est indicandus cum in pacto …) dicatur. (313vb) Schlussschrift: Consummatum fuit hoc opus in civitate Pizanensis per fratrem Bartholo meum de sancto Concordio ordinis fratrum Predicatorum doctorem decretorum. Anno domini Mo 338 die septima mensis Decembris tempore sanctissimi in Christo patris ac domini domini pape Benedicti duodecimo. Predictus autem frater Bar[tholomeus] compilator huius libri obiit anno domini Mo CCCo XLVII secunda die Iulii. Cuius anima requiescat in eo qui vivit et regnat per omnia secula seculorum. (rot umrandet:) Finitus autem est iste liber presens anno domini etc. vigesimo septimo in vigilia sancti Andree per manus Nicolai Petri hora II post meridiem vel quasi etc. (29. November 1427) 3 (314r) Forts. von Nr. 1, Bl. 1r. Inc. mut.: ] Nat cogna spiri cogna le sangwinis uxor … – Expl.: … peniteat sentencia ver post regulam iuris. 4 (314va–b) Diverse Kurztexte. 1) Tit.: Hec servare decet quemlibet hominem religiosum. – Inc.: Devotus sit in choro discretus in capitulo sobrius in refectorio disciplinatus in omni loco … – Expl.: … numquam extra se in cordis evagacione. Parallelüberlieferung: Mainz, Wiss. Stadtbibl., Hs. I 314, Bl. 120v (hier Bernardus Claraevallen sis zugeschrieben) und Hs. I 392, Bl. 205r. 152 Cod. 146 2) Tit.: Casus in quibus clericus tenetur respondere coram iudice seculari. – Inc.: Quando feudum tenet a layco de iudice c. ceterum … – Expl.: … debet actor defendere coram iudice seculari ff. de iudi[ce] l. si heres pre aliis (?). 3) Tit.: In istis casibus non tenetur quis confiteri proprio sacerdoti. – Inc.: Item si est hereticus. Item si publicus peccator pena usurarius concubinarius etc. … – Expl.: … Item si peccatum de quo vult confiteri esset commissum alteri sacerdoti etc. Schlussschrift: Hos casus ponit sanctus Thom[as] de Aqui[no] in quarto Sentenciarum di. XVII. 4) Darunter von anderer Hand: Inc.: Quoniam apostolica sedes intendit providere negociis non personis … – Expl. mut.: … sed quantum ad numerum dierum tunc habet octavas ut dicit c. preallegatum. Quoniam (?) [ 5) Tit.: Difinicio conf[irmata?] Concilii Basiliensis. – Inc.: Item ubi quis in testamento disposuit … – Expl.: … quod deest in defuncto igitur etc. 6) Darunter: Patronus est turpitudinis qui celat crimen uxoris Extra de iure iur. Quemadmodum [ (Ir–v) leer. U. S. Cod. 146 REGEL DES HL. AUGUSTINUS. HUGO DE SANCTO VICTORE (?). HUMBERTUS DE ROMANIS Perg. I, 160, I Bl. 290 × 220. Süddeutscher Raum (T. I: Neustift?), 1. H. 15. Jh. (vor 1445). B: Drei Teile: I (I, 1–5), II (6–12a[13]), III (13[14]–158[160], I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 12 und 93 doppelt angeführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. etwas abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck EX PLICAT. REGULÆ SANCTI AUGUSTIN. in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 158vb Besitzvermerk 15. Jh., radiert Iste liber est beate virginis (?). Fehlender Quinio zw. Bl. 121 und 122 gelangte 1746 in den Besitz von Samuel Szekely; vgl. Vermerk Esztergom, Kathedralbibl., Ms. II.516, Bl. 1r: Anno 1746 die 24. Martii occasione itineris in Italiam suscepti, dum in urbe Brixiensi in Tyroli diem quietis habuissemus, eo (?) tum profectus sum ad monasterium Neo-Cellense Neustifft dictum Canonicorum Regularium S. Augustini Congregationis Lateranensis, ubi praelatus est Reverendissimus ac Perillustrissi mus D. P. Antonius Steigberger, Decanus Gregorius Zeiller, Bibliothecarius Caspar Meyer. Et haec mihi Samuel Székely de Duba Locumtenenti manu propria. Wie die Lage nach Esztergom gelangte, ist nicht klar, möglicherweise – wie Ms. II.2, das einen Teil des Cod. 196 umfasst – als Geschenk von Sándor Mérey 1847; vgl. hierzu Cod. 196, Abschnitt G. Ebenso unklar bleibt, wann drei Blätter dieses einen Quinio verloren gegangen sind; vgl. T. III. Der in Neustift verbliebene Teil wurde mit Sicherheit erst nach 1746 neu gebunden und dabei auch beschnit ten: Reklamanten im Gegensatz zur heute in Esztergom aufbewahrten Lage kaum mehr sichtbar. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 G 8 (Tinte, Blei stift) und N. 146 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 146. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller VI 122. – Wretschko–Sprung 16. – R. Creytens, Les commentateurs dominicains de la Règle de saint Augustin du XIIIe au XVIe siècle. Archivum Fratrum Praedicatorum 33 (1963) 121–157, hier 126. – Neuhauser, Neustift 85. – Katalog ULB Tirol II 117, 205. – Handschriftencensus: Neustift 146 (http://www. handschriftencensus.de/18762). – E. Madas (in Verbindung mit K. Körmendy, J. Lauf, G. Sarbak), Katalog Cod. 146 153 der mittelalterlichen Handschriften in Esztergom. Die Bestände der Kathedralbibliothek und anderer Samm lungen (Fragmenta et codices in bibliothecis Hungariae VII-B). Deutsche Fassung unter Mitwirkung von A. Vizkelety. Esztergom, Budapest (in Druck). I (I, 1–5) B: Perg. an der rechten unteren Ecke abgegriffen. Bl. 2 am unteren Blattrand Nahtspuren. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + (1+II)5. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Bl. 1 einge hängtes Einzelbl., auf Bl. 2 aufgeklebt. S: Schriftraum 220 × 150, von feinen Tintenlinien gerahmt, zu einheitlich 28 Zeilen auf feiner Tintenlinierung. Zirkelstiche. Textualis formata (Textura), in der letzten Zeile Unterlänge des Minuskelbuchstabens g biswei len kalligraphisch gelängt. Bl. 4r am Blattrand Nachtrag von späterer Hand in Textualis cursiva, Bl. 5v ver einzelte Interlinearnachträge von späterer Hand. A: Rote Auszeichnungsstriche, Paragraphzeichen, Durchstreichungen, Überschriften. Zeilenfüllsel. Zwei- und dreizeilige rote, grüne und blaue Lombarden (Bl. 2r Punktverdickungen, Bl. 5r Ausläufer) mit rotem Fleu ronnée (Bl. 3v flüchtigeres blaues Fleuronnée, möglicherweise von anderer Hand). Charakteristisch im Bin nenfeld und als Besatz stilisierte Halbpalmetten, als Besatz zudem Knospen im Wechsel mit Dornen; l ängere Fadenausläufer, die in Halbpalmetten oder Fibrillen enden können. – Affinitäten zu der in beiden Bänden des Zollner-Graduales tätigen Fleuronnée-Ergänzungshand 1, vgl. z. B. Bl. 2r mit Bl. 306v2 im zweiten Gradual band: sehr ähnlich die Halbpalmetten, die wie ein Flügelpaar angeordnet sind. Bl. 1r vierzeilige ornamentale Deckfarbeninitiale mit breitem rotem Profilrahmen, auf dem der grüne, mit groben Blattranken gefüllte Buchstabenkörper aufliegt; blaues Binnenfeld mit weißen Federranken, deren Voluten Punktblüten umschließen. S. R. 1 (Ir–v) leer. (1r–5v) Regel des hl. Augustinus, dt., unvollständig (vgl. CPL Nr. 1839). Erhalten sind der Beginn von cap. 1, der Schluss von cap. 2, cap. 3–6 sowie der Beginn von cap. 7 (= letztes Kapitel). (1r–v) Tit.: Hye hebt sich an sand Augustins regel. – Inc.: Vor allen dingen ir lieben prüder habt got lieb dar nach den nächsten. Wan die zway gepot synd uns vorderleich gegeben … – Expl. mut.: … und ert in euch selbs got dez tempel ir worden seyt [ (2r–5v) Inc. mut. (Schluss cap. 2): ] so sy ye meyner bedurffen, ye saliger sy sind. Noch so sol der zarten speis girikait sy nicht uben … – Expl. mut. (Beginn cap. 7): … das sol man nicht sawmit lich übergeen, sunder mit fleiß und straffung sol man [ Zu den mhd. Übersetzungen der Augustinusregel s. G. de Smet, „Augustinerregeln“ (mhd.). Ver fasserlexikon2 I 545–550. II (6–12a[13]) B: Perg. fleckig und am rechten unteren Blattrand tw. stark abgegriffen. Buchblock beschnitten (vgl. Rand bemerkung Bl. 12r und v). Bl. 8 lose. Bl. 7 und 8 am unteren Blattrand Löcher und Nahtspuren, Bl. 12 und 12a jeweils Loch im Textfeld (ohne Fehlstelle im Text). Lagen: (I+1)8 + (1+I)11 + I12a(13). Bl. 8 und 9 ein gehängte Einzelbl., Bl. 8 heute lose, Bl. 9 auf Bl. 10 aufgeklebt. Zw. Bl. 12 und 12a fehlt wohl ein Doppelbl. S: 1) Bl. 6r–8v und Bl. 10r–11v Schriftraum 225 × 170, von feinen Tintenlinien gerahmt, zu 34 Zeilen. Bl. 11r und v deutlich über begrenzten Schriftraum hinaus beschrieben. Bastarda. Am Blattrand Notizen von unter schiedlichen Händen sowie Nachträge und Forts. der Rubriken von anderer Hand, bisweilen durch unter schiedliche Symbole im Text gekennzeichnet. 2) Bl. 9r–v Schriftraum 225 × 165, von kräftigen Tintenlinien begrenzt, zu 34 Zeilen. Rubriken wohl von gleicher Hand. Textualis. 3) Bl. 12r–12av Schriftraum 215 × 165, von feinen Tintenlinien gerahmt, zu 40 Zeilen. Forts. der Rubriken am Blattrand. Textualis. 154 Cod. 146 A: 1) Bl. 6r–8v und 10r–11v rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Interpunktions zeichen, Überschriften, Satzmajuskeln in Lombardenform, einfache Lombarden. Repräsentanten. Rubriken und Nachträge am Blattrand in Rot. 2) Bl. 9r–v rote Auszeichnungsstriche. Rubriken wohl von anderer Hand. 3) Bl. 12r–12av rote Auszeichnungsstriche, Paragraphzeichen, Initialmajuskeln, dreizeilige Lombarden mit wenig Dekor (Bl. 12r im Binnenfeld Spruchband mit V und O?). Repräsentanten. Rubriken. Knochenlöcher rot umrandet. 2 (6r–12v) ‹Hugo de Sancto Victore (?)›: Expositio in regulam beati Augustini, unvollständig (PL 176, 881–924. – Goy 457–478). (6r–7v) Inc. mut. (cap. 1): ] [D]eum ex toto corde et ex tota anima et proximum nostrum sicut nos. Necessaria est ita quia virtus concordie … – Expl. mut.: … unde et subditur. (rot:) Qui autem nichil habebant non ea querant in monasterio que nec foris habere potuerunt. [ (PL 176, 882B–886D). (8r–9v) Inc. mut. (cap. 2): ] peccati superbia. Prima est enim in peccando ultima remanet in conflictu … – Expl. mut. (cap. 3): … hostem nutrit. Sic ergo nutrienda est ut [ (PL 176, 889A–893A). (10r–v) Inc. mut. (cap. 6): ] esse vestimenta clericorum in quibus nichil notari possit … – Expl. mut.: … Non solum tacita voluntate intus concupiscencia oritur sed eciam per exteriorem [ (PL 176, 897B–899B). (11r–v) Inc. mut. (cap. 7): ] magis ad vindictam pertinet quam ad disciplinam. Caveant ergo fratres ut neminem reprehendant … – Expl. mut. (cap. 8): … temperat sentenciam cum subiungit. (rot:) Tamen si vestra tolleratur infirmitas ut hoc recipiatis quod posueratis in uno tamen loco sub communibus custodibus habete quod ponitis. (vom Druck leicht abweichend:) Nullus sibi aliquid operetur sed omnia vestra in communi fiant maiori studio et frequenciori alacritate quam si vobis singuli propria faceretis [ (PL 176, 904A–906B). (12r–v) Inc. mut. (Schluss cap. 9): ] redeundum est ad mansuetudinis lenitatem. Nam pertinax motus facile … – Expl. mut. (cap. 10): … accipere oracionis effectum. (rot:) Qui autem numquam vult petere veniam aut non ex animo petit sine causa est in monasterio eciam si inde non proii ciatur (!) [ (PL 176, 913C–916A). (12ar–v) Inc. mut.: (cap. 11) ] verbere et castigacione in ipso inicio peccati … – Expl. mut.: … Iusto non est lex posita. Ubi enim spiritus [ (PL 176, 921A–923C). III (13[14]–158[160], I) B: Vereinzelt kleine Löcher und Nahtspuren. Bl. 19 Riss. Bl. 92 am unteren Blattrand Rechteck herausgeschnit ten. Lagen: 13.V141(143) + IV149(151) + (IV+1)158(160) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Nachsatz bl. als Spiegel auf dem HD aufgeklebt. Bl. 150 eingehängtes Einzelbl., auf Bl. 151 aufgeklebt. Ab Bl. 13 am oberen Blattrand mittig zeitgenössische Foliierung in arabischen Ziffern 41–195 (mit den Folioangaben im Register übereinstimmend, allerdings wird hierbei das Verso immer zum darauffolgenden [und nicht zum vorangehenden] Recto gezählt), Blätter mit Foliierung 151–160 (= Quinio) fehlen (s. bei G). Lagenrekla manten stark be- bzw. weggeschnitten. S: Schriftraum 210 × 140/145. Zwei Spalten, von feinen Tintenlinien gerahmt, zu 35–39 Zeilen. Zirkelstiche. Zwei Hände: 1) Bl. 13ra–156ra: Bastarda. Auszüge aus der Regula Augustini in Auszeichnungsschrift (Textualis formata [Textura]) von gleicher Hand. Vereinzelt Nachträge am Blattrand und interlinear; 2) Bl. 156va–158vb (Register): Textualis, auf feiner Tintenlinierung. Zählung der Kapitel in Rot (tw. arabische Ziffern, tw. römische Zahlen). Von Hand der Foliierung Blattangaben (Numerus foliorum) in arabischen Ziffern in eigener Spalte (bis auf Bl. 158rb immer links der Titelangaben) später hinzugefügt. Am Blattrand vereinzelt Nachträge in Bastarda. A: 1) Bl. 13ra–156rb rote Auszeichnungsstriche, Überschriften (Bl. 54vb und 141va mit Repräsentant am unte ren Blattrand), Paragraphzeichen (vereinzelt blau, Bl. 49ra mit geschlungenem Ausläufer), Unterstreichungen. Als Orientierungshilfen Nota-Hinweise, X-förmige Zeichen, Haken, Zwei-, Drei- und Vierpunktgruppe. Zeigehände (meist flüchtig gezeichnet; auch mit Ärmeldraperie, z. B. Bl. 119rb und 146ra). Zwei- bis vier zeilige blaue, hell- und dunkelrote Lombarden (Bl. 131va türkisgrün), vereinzelt mit Punktverdickungen, Cod. 146 155 Aussparungen, einfachem schwarzem Fleuronnée (Bl. 84rb im Binnenfeld gegenständiges Halbpalmetten paar). Lombarden mitunter nicht ausgeführt (Bl. 82ra Vorzeichnung einer C-Lombarde). Oberlängen in der ersten Zeile und Unterlängen in der untersten Zeile bisweilen gelängt und verziert; in der obersten Zeile tw. cadellenartig, häufig mit Köpfen in Federzeichnung (z. B. Bl. 87r). Einige ungelenke Bas-de-page-Feder zeichnungen: Bl. 50r liegender Hund, Tierkopf mit Hörnerpaar; Bl. 114r langbeiniger rot-schwarzer Vogel mit Fisch im Schnabel; Bl. 75r männliche Büste (evtl. mit dem Stift ausgeführt, mit dem auch die auf Petrus Figuli bezogenen Vermerke Bl. 113va–119rb eingetragen wurden). Repräsentanten. 2) Bl. 156va–158vb (Register) rote Überschrift, Kapitelzählung. Zeigehände. Einzeilige abwechselnd rote und blaue Lombarden. Bl. 156va fünfzeilige blaue Lombarde mit rotem Fleuronnée (im Binnenfeld Knospenräder, als Besatz eng stehende Knospenreihen). Oberlängen in der ersten Zeile und Unterlängen in der untersten Zeile bisweilen gedehnt und (cadellenartig) verziert. S. R. 3 (13ra–158vb) Humbertus de Romanis: Expositio regulae s. Augustini, unvollständig (Ed.: J. J. Berthier, B. Humberti de Romanis … opera de vita regulari I. Roma 1888, ND Torino 1956, 43–633. – Kaeppeli Nr. 2016). Kapiteleinteilung in Hs. von Ed. abweichend. Der Text beginnt mit dem Ende von cap. 44; der Schluss von cap. 151, cap. 152–162 sowie der Beginn von cap. 163 fehlen. (13ra–121vb) Cap. 44–151 (= Ed. cap. 53–162). Inc. mut.: ] erectis quasi orans huius (darüber: di) exemplum … – Expl. mut.: … Quod ergo dicitur interdum quod hoc non debent facere [ Der hier fehlende Quinio (s. bei B) heute in Esztergom, Kathedralbibliothek, Ms. II 516. Aller dings sind von den urspr. fünf Doppelbl. nur mehr zwei erhalten (urspr. Foliierung: Bl. 151 und 160 sowie 154 und 157), vgl. Katalog der mittelalterlichen Handschriften in Esztergom. (122ra–156rb) Cap. 163–198 (= Ed. cap. 174–209). Inc. mut.: ] Prima est certitudo … Schlussschrift (unmittelbar an Text anschließend): Hoc additur in fine ut sciatis quicumque ordinis nostri hoc inpleverit se totam regulam implesse et ad amplius non tueri Deo gracias qui vivit et regnat in secula seculorum Amen. (156va–158vb) Register mit später hinzugefügter Blattangabe. Tit.: In nomine domini Amen. Incipit tabula super exposicionem regule beati Augusti episcopi quam exposicionem frater Humbertus magister ordinis ad utilitatem ac peticionem fratrum o rdinis Predicatorum conpilavit. – Inc.: De laudibus regule beati Augustini Capitulum primum … – Expl.: … De hiis que valent ad remissionem habendam CLXXXXVII. De temptacion ibus CLXXXXVIII. Lit.: Creytens 120–157. (158vb) Besitzvermerk (s. bei G). 4 Am Blattrand vereinzelt Notizen von anderer Hand: (113vb) am unteren Blattrand radierte Notiz Anno domini Mo CCCCo XLV frater Petrus Figuli Rat[isponiensis] (?) dyocesis fuit in monasterio Nova[cellensi] dioc[esis] Brixinensis. (114ra) am unteren Blattrand radierte Notiz Anno domini Mo CCCCo XLVo frater. (119rb) am unteren Blattrand radierte Notiz Figuli. (Ir–v) leer. U. S. 156 Cod. 153 Cod. 153 IOHANNES HEROLT (DISCIPULUS) Pap. I, 168, I Bl. 270 × 205. Wiener Raum, 4. Jz. 15. Jh. B: Pap. z. T. stark fleckig und beschädigt: Bl. 5, 49, 89 Löcher bzw. Risse, Bl. 13, 27, 61, 157 eingerissen. Buchblock beschnitten. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasser zeichenbefund. Identische Wasserzeichen: v. a. in 1437/38 datierten Hs. aus dem Wiener Raum, z. B. Wien, Schottenstift, Archiv, Hs. 280, Archiv, Hs. 280 und Hs. 381 (beide dat. 1438); Klosterneuburg, Stiftsarchiv, Grundbuch 17/2 (dat. 1437). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + (1+VI)13 + 11.VI145 + (VI–1)156 + VI168 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgek lebt. Bl. 1 eingefügtes Einzelbl., auf Bl. 2 aufgeklebt, urspr. letztes Bl. einer Lage (vgl. Reklamante). Gegenbl. zu Bl. 146 herausgeschnitten (Textverlust). Erstes bzw. letztes Blatt der Lage jeweils durch einen aufgeklebten Papierfalz verstärkt. Zw. Bl. 156 und 157 Pergamentfalz. Lagenreklamanten, z. T. beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 115 doppelt angeführt, 116 ausgelassen. S: Schriftraum 205 × 145/150. Zwei Spalten, von feinen Tintenlinien gerahmt, zu 37–41 Zeilen. Bastarda. A: Rote Auszeichnungsstriche, Unterstreichungen, Über- und Schlussschriften. Zwei- bis siebenzeilige rote Lombarden, tw. mit Punktverdickungen. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck MANUSCRIPT. in Goldrahmung, im dritten Signatur schild (s. bei G). Kapitale mit naturfarbenem und blauem Zwirn umstochen. Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signatur der ULB Tirol II 2 H 1 (Tinte, Bleistift) und Nr. 153 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signatur schild der ULB Tirol 153. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 133, 205. 1 (Ir–v) leer. (1ra–160vb) ‹Iohannes Herolt (Discipulus)›: Sermones de tempore (Schneyer II Nr. 11?–91). (1ra) Vermutlich Dnca 4. adv. (Schneyer II Nr. 11?). Inc. mut.: ] miracula et in hoc apparebit ipsorum consciencia unde Gregorius … – Expl.: … Ite maledicti in ignem eternum qui paratus est dyabolo et angelis eius a quo nos custodiat dominus noster Iesus Christus qui vivit et regnat in secula seculorum Amen. (1va) Tit.: Sequitur sermo de nativitate Christi undecimus bonus et utilis (Schneyer II Nr. 12). – Inc.: Ecce ewangelizo vobis … Luc. 2o (10–11). Nota quod inter omnia festa letabunda tocius anni hoc festum nativitatis est tanto lecius … – Expl.: … non licet comedere carnes in die nativi tatis Christi. Extra de observancia ieiunii (?). (4va) Tit.: Item sermo de nativitate Christi delusoribus. Sermo XIIus bonus (Schneyer II Nr. 13). – Inc.: Anuncio vobis gaudium magnum … Luc. 2o (10). Scire debetis in Christo dilectissimi quod dies hodierna ex salvatoris nostri nativitate nobis redditur gaudiosa … – Expl.: … animas vero in penas eternas perpetuo secum cruciandas detrudendo etc. (7rb) Tit.: De sancto Stephano. Sermo tredecimus (Schneyer II Nr. 14, Bibelzitat abweichend, vgl. Nr. 15). – Inc.: S (!) Stephanus vidit celos apertos. Actuum 7o (56). Aristoteles primo Ethi corum finis per quem omnia desiderantur et sperantur optimus est … – Expl.: … ubi nullus ordo sed sempiternus horror inhabitat etc. (9rb) Tit.: Divisio sermonis huius (Schneyer II Nr. 15). – Inc.: Ecce video celos apertos (Act 7,56). Prosequendo sermonem iam inceptum tunc sciendum quod quanto celum aperitur per intensam contricionem … – Expl.: … et beato Stephano inmense gracie referuntur etc. Cod. 153 157 (11va) Tit.: Sermo quindecimus de sancto Iohanne ewangelista (Schneyer II Nr. 16). – Inc.: Se quere me. Iohannis 21 (19). Verba proposita sunt a Christo beato Petro dicta et iam dicuntur cuilibet homini qui vult salvari … – Expl.: … quia ambo apud Deum sunt equales meriti etc. (14ra) Tit.: De parentibus qui pueros occidunt. Sermo de Innocentibus 16us XVIus (Schneyer II Nr. 17). – Inc.: Mittens Herodes occidit omnes pueros … Mt. 2o (16). Ex quo hodie peragitur festum illorum puerorum innocentium qui ab iniquo Herode interfecti sunt … – Expl.: … Salomon dicit: Qui parcit virge odit puerum suum etc. Sequitur. (15vb) Tit.: Dominica infra octavas nativitatis Christi. Sermo XVIIus quare parvulus factus est (Schneyer II Nr. 18). – Inc.: Puer autem crescebat … Luc. 2o (40). Ex quo in presenti ewangelio fit mencio de puero nobis recenter nato. Tunc pro presenti sermone tria sunt dicenda. Primo quare Christus parvulus factus est … – Expl.: … et ymaginem regina mihi tribuit etc. (17va) Tit.: Dominica. Sermo XVIII quod impedimenta perfectus (!) sunt septem (Schneyer II Nr. 19). – Inc.: Puer autem crescebat etc. (Lc 2,40). Sicut puer Ihesus crevit corporaliter ita et nos crescere debemus spiritualiter … – Expl.: … iniuria esset minima tunc esset veniale. (19rb) Tit.: In die circumcisionis domini. Sermo XIXus bonus (Schneyer II Nr. 20). – Inc.: Postquam consummati sunt dies octo … Luc. 2o (21). Solent homines in novo anno munuscula sibi invicem dare … – Expl.: … ipsi dyaboli maxime repellerentur per hoc nomen Iesus etc. (21ra) Tit.: Iuvenes debent a iuventute servire Deo. Sermo XXus (Schneyer II Nr. 21). – Inc.: Postquam consummati sunt dies octo etc. Luc. 2o (21). Sciendum quod Christus die octavo cir cumcisus est et sanguinem suum fudit … – Expl.: … et fideliter Deo servierunt etc. (23ra) Tit.: Sequitur sermo XXIus in festo epiphanie domini (Schneyer II Nr. 22). – Inc.: Ecce magi veniunt … Mt. 2o (1). Hodierne diei festivitas magna est. Unde merito dicitur apparicio … – Expl.: … Gregorius. Mala que nos hic premunt ad Deum ire compellunt etc. (25vb) Tit.: De eodem festo de ioco. Sermo XXIIus (Schneyer II Nr. 23). – Inc.: Videntes stellam gavisi sunt gaudio magno valde etc. (Mt 2,10). Glosa non fuit satis ewangeliste dicere gavisi sunt sed addit valde magno gaudio … – Expl.: … ad inferos transmissi sic et illi accidit etc. (27va) Tit.: Dominica infra octavas epiphanie sermo XXIIIus (Schneyer II Nr. 24). – Inc.: Cum factus est Iesus annorum duodecim … Luc. 2o (42). In presenti enim ewangelio instruit Maria parentes qualiter se debeant habere ad filios … – Expl.: … et Deo fideliter servivit et sic salvatus est etc. (30va) Tit.: De eadem dominica. Ffilii tenentur VI parentibus. Sermo XXIIIIus (Schneyer II Nr. 25). – Inc.: Descendit cum eis … Luc. 2o (51). Augustinus Non solum Christi verba sunt nobis in documentum sed eciam Christi facta sunt nobis in exemplum … – Expl.: … vidit illum Parisius cum aliis multis pueris et hominibus etc. (32va) Tit.: Dominica prima post octavas epiphanie. Sermo XXVus (Schneyer II Nr. 26). – Inc.: Nupcie facte sunt … Ioh. 2o (1–2). Quamvis heretici miseri menciantur carnale martirium esse illicitum eorum tamen falsitas manifeste apparet … – Expl.: … eterna dampnatio a qua nos cu stodiat dominus noster Iesus Christus A. (35va) Tit.: Sermo XXVIus quere a quibus non prospere succeditur in matrimonio (Schneyer II Nr. 27). – Inc.: Nupcie facte sunt in Chana Galilee etc. (Io 2,1). Ex quo ewangelium hodiernum mencionem facit de matrimonio et de nupciis in quibus Christus Deus et homo cum benedicta matre … – Expl.: … Quod audiens Iohannes admirans gracias Deo egit etc. (37va) Tit.: Defectus quos Christus assumpsit. Dominica 2a. Sermo XXVIIus (Schneyer II Nr. 28). – Inc.: Cum descendisset Iesus … Mt. 8 (1). Sciendum quod dicit Haymo Mons in quo Deus sedet celum intelligitur … – Expl. (von Schneyer II abweichend): … efficitur servus dyaboli nec propria voluntate in q. 1 Non est. (39rb) Tit.: De honore sermo XXVIIIus de eadem dominica (Schneyer II Nr. 29). – Inc.: Ego veniam et curabo eum. Mt. 8 (7). In hiis verbis notatur magna humilitas Christi qui servum cen turionis dignatus est visitare … – Expl. (von Schneyer II abweichend): … in tenebras exteriores etc. Hoc est dicendum de penis inferni. Illum sermonem quere infra 125 Q. Sequitur. 158 Cod. 153 (41va) Dnca 4. post epiph. Tit.: De temptacionibus. Sermo XXVIIIIus (Schneyer II Nr. 32). – Inc.: Motus magnus factus est … Mt. 8 (24). Mr. 4 (vgl. Mc 4,37). Luc. 8 (vgl. Luc. 8,23). Ubi scien dum est quod per mare intelligitur presens mundus et per motum magnum … – Expl.: … et ita sanus frater ille discessit etc. Sequitur sermo. (44va) Dnca 5. post epiph. Tit.: Sermo XXXus quomodo boni et mali sunt permixti in hoc mundo (Schneyer II Nr. 33). – Inc.: Sinite utraque crescere … Mt. 13o (30). Sciendum quod in celo sunt solummodo boni in inferno solummodo mali … – Expl. (gegenüber Schneyer II erweitert, vgl. Schneyer II Nr. 34): … ad vitam salutis. De ewangelio eodem Triticum congregate … est dicen dum de novem gaudiis celi. Istum sermonem quere infra 83 K exemplum. (46vb) Tit.: Dominica quinta de humilitate. Sermo XXXIus (Schneyer II Nr. 35). – Inc.: Discite a me quia mitis sum … Mt. XI (29). Aristoteles In sempiternis nichil violentum. Qui ergo alte in celo vult … – Expl.: … Discite a me quia mitis sum et humilis corde etc. (48rb) Dnca in Septuages. Tit.: De dignitate hominis. Sermo XXXIIus. Dominica in Septuagesima (Schneyer II Nr. 36). – Inc.: Simile est regnum celorum homini patrifamilias … Mt. 2o (recte: 20,1). Scire debetis in Christo dilectissimi quod sancta mater ecclesia katholica hodierna die duo nobis tempora representat … – Expl.: … quanto spiritu confortabantur tanto corpore infirma bantur etc. (50ra) Tit.: De dignitate hominis sermo XXXIIus (!). Dominica in LXXa (Schneyer II Nr. 37). – Inc.: Simile est regnum celorum homini patrifamilias … Mt. XXo (1). Ex quo in presenti ewan gelio fit mencio de patrefamilias qui conduxit operarios … – Expl.: … animam illius ancille ad celum deduxit etc. (51vb) Tit.: Dominica in LXa de creacione hominis. Sermo XXXIIII bonus (Schneyer II Nr. 38). – Inc.: Exiit qui seminat … Luc. 8 (5). Ubi sciendum quod seminator est Deus omnipotens qui exiit seminare semen suum id est genus humanum creando … – Expl.: … Cupio dissolvi et esse cum Christo etc. (54ra) Tit.: Dominica in LXa de verbo Dei. Sermo XXXVus bonus (Schneyer II Nr. 39). – Inc.: Semen est verbum Dei. Luc. 8 (11). Est enim semen verbum Dei quod Christus de celo attulit in terra seminavit … – Expl.: … in locis penalibus iusticia dampnantis dampnat etc. (55rb) Tit.: Dominica in Quinquagesima de gaudio mundano. Sermo XXXVIus (Schneyer II Nr. 40). – Inc.: Ecce ascendimus Ierosolimam … Luc. 18 (31–32). Mt. 2o (vgl. recte Mt 20,18– 19). Marci 10 (vgl. Mc 10,33–34). Duo enim manifestantur in ewangelio presenti scilicet passio Christi et illuminacio cuiusdam ceci … – Expl. (von Schneyer II abweichend): … et facta est postmodum mulier sancta etc. Sequitur. (58ra) Tit.: Sermo XXXVII de corea et de eadem dominica (Schneyer II Nr. 41). – Inc.: Tradetur enim gentibus … Luc. 18 (32). Ubi sciendum quod passio Christi que in hodierno ewangelio nobis proponitur retrahere nos debet ab illicitis gaudiis huius seculi … – Expl.: … mundus non sit diligendus illum sermonem infra 136 ff. Secuntur sermones Quadragesime etc. (60va) Tit.: Incipit Quadragesimale in die cinerum fiunt sex. Sermo XXXVIIIus (Schneyer II Nr. 42). – Inc.: Convertimini ad me … Iohel 2o (12). Unde Iohannes de Sancto Geminiano Vide mus enim quod cum animal brutum sentit se wlneratum sollicite querit et procurat sanacionis remedium … – Expl.: … a quibus tandem suffocatus penam irrisionis luit etc. (62va) Tit.: Dominica prima in Quadragesima de ieiunio. Sermo XXXVIIIIus (Schneyer II Nr. 43). – Inc.: Cum ieiunasset quadraginta diebus … Mt. 4o (2). Mr. 1o (vgl. Mc 1,13). Luc. 4o (vgl. Lc 4,1). Christus ieiunavit nobis in doctrinam et exemplum quia secundum Gregorium Omnis Christi actio nostra est instructio … – Expl.: … pane et aqua in uno peccato mortali ieiunaret etc. (65rb) Tit.: Dominica prima in Quadragesima de septem temptacionibus. Sermo XLus (Schneyer II Nr. 44). – Inc.: Et accedens temptator dixit ei Si filius Dei es … Mt. 4o (3). Sciendum quod Christus voluit temptari ut ostenderetur quod nos spiritualiter pugnare oporteret … – Expl.: … est ordo monachorum. Et statim egressus factus sum monachus etc. Cod. 153 159 (67va) Bei Schneyer II ohne eigene Nr. (direkt an Nr. 44 anschließend). Tit.: Ffidem falsificant 24 genera hominum. Sermo XLIus dominica secunda. – Inc.: Mulier magna est fides tua … Mt. 15o (28). Mr. 7o (vgl. Mc 7,25) etc. Aristoteles 8 Phisicorum Illud est prius quo non existente non sunt alia sed (radiert: sum?) sine fide … – Expl. (von Schneyer II abweichend): … et in punctu ad inferna descendunt etc. (69ra) Tit.: Dominica tercia in Quadragesima de iuramentis. Sermo XLIIus (Schneyer II Nr. 45). – Inc.: Erat Iesus eiciens … Luc. XIo (14). Mt. 22o (vgl. recte Mt 9,32–33). Mr. 3o (22). Iohannes de Sancto Geminiano Videmus naturaliter quod lux habet expellere tenebras. Christus autem est lux … – Expl.: … et Deus sibi illud peccatum indulsit etc. (71vb) Tit.: Dominica quarta in Quadragesima scilicet media de contricione et confessione. Sermo XLIIIus (Schneyer II Nr. 46). – Inc.: Est puer unus hic qui habet quinque panes … Ioh. 6to (9). Mt. 14 (vgl. Mt 14,17). Iohannes de Sancto Geminiano dicit quod totum officium presentis diei dominice continet quasi quedam nova et leta … – Expl.: … nomine de cetero non oblatrare etc. (74va) Tit.: De penitencia vera et falsa. Sermo XLIIIIus de eadem dominica (Schneyer II Nr. 47). – Inc.: Penitenciam agite … Mt. 3o (2). Aristoteles 5o Phisicorum Omnis motus tendit ad aliquem terminum per debita media … – Expl.: … quia misericordia opera eius. Et de ista materia quere exempla infra 2 N. item 14 ff. Item 43 Q. (77ra) Tit.: Sequitur dominica de passione domini. Sermo XLVus (Schneyer II Nr. 48). – Inc.: Tulerunt ergo lapides … Ioh. 8 (59). Aristoteles 8 Ethicorum dicit Vera (gestrichen: iusticia) amicicia habet reamacionem. Et quia Christus vere nos dilexit … – Expl.: … et laudabiliter post modum vixit et in domino quievit. Quod et nobis concedat Ihesus Christus Amen etc. (79ra) Unmittelbar an Sermo XLV anschließend: De eadem dominica in passione domini alius sermo. Qui ex Deo est verba Dei audit. Istum sermonem quere ante 35 V (vgl. Schneyer II Nr. 49). (79rb) Tit.: Dominica palmarum de indigne communicantibus. Sermo XLVIus (Schneyer II Nr. 50). – Inc.: Benedictus qui venit in nomine domini. Mt. 21 (9). Mr. XI (9). Luc. 19 (recte: 13,35). Ioh. 12 (13). Hodie sancta mater ecclesia duas processiones recolit … – Expl.: … absti nendi et emendandi vitam suam etc. (81rb) Tit.: In cena domini de percepcione sacramenti eukaristie. XLVIIus (Schneyer II Nr. 51). – Inc.: Probet autem seipsum homo … Prima Cor. XI (28). Aristoteles 2o De anima Simile a simili nutritur sicut patet in cibo digesto a quo corpus nutritur … – Expl.: … post confessionem et penitenciam digne communicavit. (84ra) Unmittelbar an Sermo XLVII anschließend (Schneyer II Nr. 52). Inc.: Item pro 2o sermone in hac die tria sunt dicenda. Primo quomodo 4or genera hominum … – Expl.: … oravit quinque Pater noster. Hoc quere infra sermone 80 V. (84ra) Tit.: In die parascevensi duodecim fructus de memoria passionis Christi. Sermo XLVIIIus (Schneyer II Nr. 53). – Inc.: O vos omnes qui transitis … Trenorum primo (Lam 1,12). Unde Iohannes de Sancto Geminiano dicit Conveniens est ut senciat (!) membrum quod sentit caput … – Expl.: … Interior est planctus gravior cum te tam ingratum experior etc. (86ra) Tit.: In die parasceves de passione Christi. Sermo XLIXus (Schneyer II Nr. 54). – Inc.: Inclinato capite emisit spiritum … Ioh. 19 (30). Iohannes de Sancto Geminiano dicit Mors sal vatoris karissimi quam hodie recolimus fuit ultima consummacio dominice passionis … – Expl.: … et post mortem celos introivit. Hoc et nobis prestare dignetur Iesus Christus virginis Marie filius Amen. (89rb) Vermerk (Schneyer II Nr. 55): Item tercium sermonem de passione scilicet quomodo Christus assumpsit decem defectus humane nature. Quere ante 27o sermone A. (89rb) Weiterer Vermerk (nicht bei Schneyer II): Item exemplum bonum de passione domini de milite illo qui quinque Pater noster oravit. Quere infra sermone 80 V. 160 Cod. 153 (89va) Tit.: Sermo in festo pasce. Sermo Lus (Schneyer II Nr. 56). – Inc.: Hec est dies q[uam] fecit dominus … Ps. 117us (24). Sciendum quod presens festivitas precellit alias festivitates in sollemp nitate et iocunditate … – Expl.: … et sic fatue redierunt ad suas sorores etc. (91vb) Tit.: Signa digne communionis. Sermo LIus (Schneyer II Nr. 57). – Inc.: Mane nobiscum domine. Luc. 24 (29). Hec verba canit ecclesia per illud sacrum tempus pascale usque ad ascen sionem … – Expl.: … Et hec est caritas sufficiens sed ducit ad nimiam gloriam. (94va) Tit.: In festo pasce de recidivacione. Sermo LIIus (Schneyer II Nr. 58). – Inc.: Mane no biscum domine. Luc. 24 (29). In quibus verbis monemur ne a bono iam incepto desistamus … – Expl.: … ceteri penitenciam egerunt ad salutem etc. (96ra) Tit.: Item in festo pasce. Sermo LIIIus (Schneyer II Nr. 59). – Inc.: Mane nobiscum domine. Luc. 24 (29). Circa quod sciendum quod sunt sex que debent hominem retrahere ab omni peccato mortali … – Expl. (von Schneyer II abweichend): … et circa me perpendo et considero quomodo gaudere possum undique cum hoc seculo etc. (99rb) Tit.: De eadem feria aliqui sunt modis stulti aliqui modis sapientes. Sermo LIIIIus (Schne yer II Nr. 60). – Inc.: O stulti et tardi corde ad credendum. Luc. 23 (recte: 24,25). Sciendum quod 2m dictum Salomonis stultorum infinitus est numerus … – Expl.: … si usque hodie peccent vel peniteant etc. (101ra) Tit.: Quarta feria pasce de laboratoribus. Sermo LVus (Schneyer II Nr. 61). – Inc.: Vado piscari. Ioh. 21o (3). Ex quo hodie est ultima dies celebris presentis festivitatis pasce et modo homines redire debent ad pristinos labores suos … – Expl. (von Schneyer II abweichend): … ab eo noluit exire sed eius verba contempsit etc. Quere hoc exemplum sermone 26 V. (103va) Nicht bei Schneyer II, jedoch im Druck GW 12346. Tit.: Dominica in octava pasce. Sermo LVIus. Decem confert societas bona. – Inc.: Thomas autem non fuit cum eis quando venit Iesus. Ioh. 20 (24). Notandum quod beatus Thomas Christum discipulis apparentibus non vidit quia non fuit cum eis sed post octo dies eum vidit … – Expl. (= Expl. Schneyer II Nr. 61): … secundum alterius voluntatem et ideo maioris virtutis sunt etc. (104va) Tit.: Sermo LVIIus de cycatricibus wlnerum (Schneyer II Nr. 62). – Inc.: Inffer digitum tuum huc. Iohannis 20 (27). Sciendum primo quod Christus servavit cycatrices wlnerum in suo corpore glorificato … – Expl.: … sed dignitas et pulchritudo erit etc. (106ra) Tit.: Sermo LVIIIus de recidivantibus de eadem dominica (Schneyer II Nr. 63). – Inc.: Respondit Thomas dixit Deus meus et dominus meus. Ioh. 20 (28). Verba predicta quilibet potest dicere qui digne in hoc sacro tempore communicavit … – Expl.: … octo genera hominum vix vel raro se emendant etc. (107vb) Tit.: Dominica prima post octavas pasce de prelatis et subditis. Sermo LVIIIIus (Schneyer II Nr. 64). – Inc.: Ego sum pastor bonus. Ioh. decimo (11). Sciendum quod per pastorem in pre senti ewangelio intelliguntur prelati et per oves intelliguntur subditi … – Expl.: … tales eciam tenentur ad restitucionem. (109vb) Tit.: De eadem dominica (Schneyer II Nr. 65). – Ego sum pastor bonus (Io 10,11). Hic Christus seipsum commendat. Unde hic est dicendum quando quis potest se licite commendare et quando est veniale et quando est mortale hoc quere infra sermone 107 K. (109vb) Tit.: De eadem dominica (Schneyer II Nr. 66). – Animam meam pono pro ovibus meis (Io 10,15). Hic dicendum est de caritate Dei quam habet ad nos et eciam de caritate quam nos ad ipsum habere debemus. Istum sermonem quere infra 119 P. (109vb) Tit.: Dominica secunda post octavas pasce quare antiqui patres tam diu vixerunt. Sermo LXus (Schneyer II Nr. 67). – Inc.: Modicum et iam non videbitis me. Ioh. 16 (16). Hec verba lo cutus est dominus Deus et ante passionem suam post cenam … – Expl.: … mortuus est. Et alius dixit Certe tibi ludum dimitto etc. (112vb) Tit.: De eadem dominica de partu mulierum. Sermo LXIus (Schneyer II Nr. 68). – Inc.: Mulier dum parit tristiciam habet. Ioh. 16 (21). Ex quo in verbis premissis fit mentio de partu mulierum. Tunc in presenti sermone tria sunt dicenda … – Expl.: … quia de porco fuit nutritus. Cod. 153 161 (115rb) Tit.: De eadem dominica (Schneyer II Nr. 69). – Vos autem contristabimini (Io 16,20). Hic est dicendum quare Deus permittit aliquos bonos homines ita acerbissime tribulari in pre senti. Istum sermonem quere ante 29 O. (115va) Tit.: De eodem (Schneyer II Nr. 70). – Mundus autem gaudebit (Io 16,20). Hic est di cendum quare gaudium mundi est fugiendum et quando est mortale. Istum sermonem quere ante 36 D. (115va) Tit.: Dominica tercia post octavas pasce. Isti arguuntur a Deo qui factis ostendunt se non credere in Deum. Sermo LXIIus (Schneyer II Nr. 71). – Inc.: Arguet mundum de peccato quia non credunt in me. Ioh. 16 (8). Sciendum quod plura genera hominum sunt qui dicunt verbis se credere in Deum … – Expl.: … utilem ad custodiam sui ad augmentum meriti etc. (117vb) Tit.: De eadem dominica. Sermo LXIII. De correctione (Schneyer II Nr. 72). – Inc.: Arguet mundum de peccato. Ioh. 16 (8). Sciendum quod sicut Deus arguet peccatores de suis excessibus in futuro … – Expl.: … in suis missis et oracionibus quia valde affligor etc. (119vb) Tit.: De eodem (Schneyer II Nr. 73). – Arguet mundum de peccato (Io 16,8). Glosa sem per quod habet. Unde hic est dicendum quomodo quis committit 7m peccata mortalia quere infra sermone 141 I. (119vb) Tit.: De eodem (Schneyer II Nr. 74). – Arguet mundum de iusticia (Io 16,8). Glosa sem per quam non habet. Unde hic est dicendum quomodo homo debet iustus inveniri si wlt salvari. Istum sermonem quere infra 154 S. (119vb) Tit.: Dominica quinta scilicet rogacionum de oracione. Sermo LXIIIIus (Schneyer II Nr. 75). – Inc.: Petite et accipietis. Ioh. 16 (24). Benignitas salvatoris Dei nostri maior est in exaudiendo peticiones filiorum suorum … – Expl.: … quia providet illis meliora quam petunt etc. (123rb) Tit.: De eadem dominica de Pater noster. Sermo LXVus (Schneyer II Nr. 76). – Inc.: Petite et accipietis. Ioh. 16 (24). Ex quo dominus noster Iesus Christus in verbis premissis invitat nos ad orandum tunc hic queritur … – Expl.: … sed quam caritas fraternalis commendat etc. (126ra) Tit.: In festo ascensionis. Sermo (gestrichen: LXus) LXVIus (Schneyer II Nr. 77). – Inc.: Postquam locutus est eis Assumptus est in celum … Mr. 16 (19). Sciendum quod ab origine mundi non fuit tale festum in celo nec unquam usque post iudicium erit … – Expl.: … si vis plura dicere de baptismo quere infra sermone 77 B. (127ra) Tit.: De eodem festo. Signa veri Christiani. Sermo LXVIIus (Schneyer II Nr. 78). – Inc.: Signa autem eos qui crediderint. Hec sequentur. Mr. ultimis (16,17). Quamquam nesciatur de homine quoquam an salvetur vel dampnetur … – Expl.: … et sermonem confirmante sequentibus signis etc. (129va) Tit.: Sermo infra octavas ascensionis de preparacione ad graciam. Sermo LXVIIIus (Schneyer II Nr. 79). – Inc.: Cum venerit paraclitus quem ego mittam vobis a patre. Ioh. 15 (26). Sicut in dominica proxima futura recolitur spiritus sancti missio … – Expl.: … sed in deserto deficiente eis farina receperunt manna etc. (131vb) Tit.: De eadem dominica de excommunicacione. Sermo LXVIIIIus (Schneyer II Nr. 80). – Inc.: Absque sinagogis facient vos. Ioh. 15 (recte: 16,2). Sciendum quod sicut Iudei olim tenue runt se ad discipulos Christi scilicet separando eos a participacione et communione ipsorum … – Expl.: … in ecclesiam posuit statim moriebatur etc. (134va) Tit.: De festo pentecostes de spiritu sancto. Sermo LXXus (Schneyer II Nr. 81). – Inc.: Ad eum veniemus et mansionem aput eum faciemus. Iohannis 14 (23). Sciendum quod hodierna festivitas inter alia festa dignitatem singularem obtinet … – Expl.: … et finem suum in bono terminavit quod et nobis concedat Ihesus Christus Amen. (137ra) Tit.: Alius sermo de spiritu sancto et est LXXIus (Schneyer II Nr. 82). – Inc.: Repleti sunt omnes spiritu sancto. Act. 2us (4). Aristoteles 9o Methaphisice Contraria non fiunt in eodem. Cum ergo spiritus sanctus intrat in cor hominis tunc mala et peccata depellit … – Expl.: … tria in proficientibus et tria in perfectis etc. 162 Cod. 153 (139ra) Tit.: Alius sermo quare Deus subtrahit graciam homini LXXIIus (Schneyer II Nr. 83). – Inc.: Spiritus meus super mel dulcis. Ecci. 24 (Sir 24,27). Ex quo spiritus sanctus est fons tocius dulcedinis tunc queritur quare Deus quandoque dilectis suis subtrahit in presenti consuetam graciam … – Expl.: … delevit penam et culpam propterea risi etc. (141rb) Tit.: De donis spiritus sancti. Sermo LXXIIIus (Schneyer II Nr. 84). – Inc.: Repleti sunt omnes spiritu sancto. Act. 2o (4). Ex quo iam illis sacris diebus peragitur missio spiritus sancti tunc aliquid de donis spiritus sancti pro presenti sermone dicendum est … – Expl.: … si carnali delectacione cor maculas etc. (144rb) Tit.: Fferia 4ta pentecostes Deus vocat hominem multipliciter. Sermo LXXIIIIus (Schneyer II Nr. 85). – Inc.: Nemo potest venire ad me nisi pater meus traxerit eum. Ioh. 6 (44). Sciendum quod circa predicta verba tria sunt dicenda. Primo quomodo Deus trahit et vocat hominem … – Expl.: … perseveravit in bono sine cessacione quod et nobis prestare dignetur Iesus Christus Marie filius Amen etc. (147va) Tit.: In festo trinitatis. Sermo LXXVus (Schneyer II Nr. 86). – Inc.: O altitudo diviciarum sapiencie et sciencie Dei … Ro. XI (33 und 36). Sciendum quod hodie festum trinitatis institutum est ut omnes fideles agnoscant unam simplicem deitatem et trinitatem … – Expl.: … quam sit verum quod puer dixit recognovit etc. (150va) Tit.: Item de trinitate de baptismo. Sermo LXXVIus (Schneyer II Nr. 87). – Inc.: Nisi quis renatus fuerit ex aqua … Ioh. 3o (5). Sciendum quod triplex est nativitas naturalis spiritualis et celestis … – Expl.: … inveniens sibi celum semper apertum. (152va) Tit.: Item de trinitate de signis ante et post baptismum. Sermo LXXVIIus (Schneyer II Nr. 88). – Inc.: Nisi quis renatus fuerit ex aqua et spiritu sancto etc. ubi supra. (Io 3,5). Ex premissis patet quod quam cito homo moritur qui adhuc stat in innocencia baptismali … – Expl.: … ut excusacio de ignorancia habetur. (154ra) Tit.: In festo corporis Christi. Sermo LXXVIIIus (Schneyer II Nr. 89). – Inc.: Caro mea vere est cibus … Ioh. 6 (55). Sicut dicit Aristoteles 4o Ethicorum Terribilissimum omnium terribilium videtur esse mors … – Expl.: … quia Ihesus venit et inter hec verba migravit ad dominum etc. (156vb) Tit.: Item sermo LXXVIIIIus de corpore Christi (Schneyer II Nr. 90). – Inc.: Caro mea vere est cibus … Ioh. 6 (55). Sic legimus quod unigenitus Dei filius volens [ (Textverlust, s. bei B) (157ra) Forts. Inc. mut.: ] stat circa magnum sudat in toto corpore igitur magnus ignis in Christi pectore erat qui toto corpore sudavit etc. … – Expl.: … subito salvos facit sperantes in se etc. (158va) Tit.: Item de corpore Christi isti manducant corpus Christi spiritualiter. Sermo LXXXus (Schneyer II Nr. 91). – Inc.: Qui manducat hunc panem vivet in eternum. Ioh. 16 (recte: 6,58). Loquendo de manducacione huius venerabilis sacramenti sciendum quod hoc sacramentum tri pliciter sumitur … – Expl.: … quem in cimiterio sepelierunt ubique tam grande miraculum diwl gentes etc. Schlussschrift: Explicit primus liber. 2 (161ra–b) Exemplum: Pater, filius et asinus (vgl. B. E. Perry, Aesopica: a series of texts relating to Aesop or ascribed to him or closely connected with the library tradition that bears his name I. Urbana Illinois 1952, Nr. 721. J. T. Welter, La tabula exemplorum secundum ordinem alphabeti. Recueil d’exempla compilé en France à la fin du XIIIe siècle. Paris, Toulouse 1926, Nr. 265). Tit.: Nota exemplum. – Inc.: Item mundus assimilatur duobus viris scilicet patri et filio quorum unus scilicet pater equitabat super azinum et filius pedester ambulabat … – Expl.: … si est liberalis dicunt esse fatuum. Ergo non est gaudendum de martyrio temporaliter sed tantum spiritualiter etc. (161v–168r) nur Rahmung. (168v) Federprobe: Oswald. (Ir–v) leer. U. S. Cod. 154 163 Cod. 154 MISSALE SPECIALE (TEILMISSALE) Perg. I, 47, I Bl. 295 × 195. Umfeld Neustift, 12. Jh. (mit Nachträgen 13. Jh.). B: Dickes Perg. abgegriffen, fleckig, löchrig, Nahtspuren. Lagen: 1I (Vorsatzbl., Pap. 18. Jh.) + (IV–3)5 + (IV– 1)12 + 4.IV44 + 347 + 1I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zu Bl. 1 (Textverlust) und zu Bl. 9 herausge schnitten. Zw. Bl. 3 und 4 (Lagenmitte) Verlust eines Doppelbl. (Textverlust). Bl. 45–47 drei aufeinander geklebte Einzelbl. (ohne Textverlust) als Rest einer Lage. Schluss verloren. Ab Lage 2 am Lagenende Kustoden III–VII in römischen Zahlen, stark beschnitten. Am Beginn Verlust zweier Lagen. Aufgrund des einheitlichen Schriftraums der gesamten Hs. ist die Zugehörigkeit der ersten Lage trotz anderen Schreibers und fehlender Zählung anzunehmen. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Bl. 1r–4r Schriftraum 215 × 125, seitlich von kaum sichtbaren zweifachen Tintenlinien gerahmt, zu 24 Zeilen auf kaum sichtbarer Tintenlinierung. Karolingische Minuskel. Bl. 4ra–b Seitenmitte und Zusätze am Rand von anderer Hand, wie Bl. 4va–5vb. Bl. 4va–5vb Schriftraum 215 × 125. Zwei Spalten, seitlich von zweifachen Tintenlinien gerahmt, zu 43–46 Zeilen auf Tintenlinierung. Textualis 13. Jh. (möglicherweise in Neustift hinzugefügt). Bl. 6r–47v Schriftraum 215 × 125/130, seitlich von zweifachen Tintenlinien gerahmt, zu 24 Zeilen auf Tintenlinierung. Karolingische Minuskel von anderer Hand 12. Jh. A: Rote Unterstreichungen, Zeilenfüllsel (Bl. 5rb), Benediktionskreuze, Überschriften, Satzmajuskeln in Lombardenform und zweizeilige Lombarden. Bl. 9v fünfzeilige Initialligatur VD (= Vere dignum), Bl. 10r siebenzeilige Initiale T mit flankierenden rot(-tintenfarbenen) Halbpalmetten. Zw. Bl. 9 und 10 urspr. vor handenes Kanonbild herausgeschnitten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. VD und HD fleckig und zerkratzt. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck MISSALE in Goldrahmung, im vierten Feld gelbes Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen, Bleistiftvermerk (s. bei G) und eingeklebter Papierzettel (20. Jh.) mit kurzer Inhaltsangabe, Spiegel des HD leer. G: Unsicher, ob in Neustift entstanden. Nachträge in Textualis 13. Jh. (Bl. 4r, untere Blatthälfte–5vb) mögli cherweise in Neustift eingetragen. Hs. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neu stift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 2 (Tinte, Bleistift) und Nr. 154 (Tinte) sowie Bleistiftvermerk (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 154. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 133, 205. – Baroffio 324, Nr. 16712. Missale speciale (Teilmissale). Die Hervorhebung der hl. Margaretha spricht für eine Verwendung in einer der Margarethenkirchen im Neustifter Seelsorgebereich, z. B. in Neustift selbst, Schabs, Völs am Schlern (Obervöls), Margen bei Terenten, bes. aber in Montal wegen der zusätzlichen Hervorhebung von Matthias und Matthäus (vgl. die Ausmalung der Kirche im 19. Jh.). Zu Montal vgl. das wohl in Neustift für Montal geschriebene „Teyser-Missale“ von 1414 (Innsbruck, ULB Tirol, Cod. 563), mit roter Hervorhebung der Kirchen patronin Margaretha im Kalendar, s. Katalog ULB Tirol VI 197–203 und W. Neuhauser, Das sog. Teyser-Missale (Cod. 563 der Innsbrucker Universitätsbibliothek). Der Schlern 81/7 (2007) 40–47. Ohne Notation. 1 (Ir–v) leer. (1r–4r) Sequenzen (weitgehend wie Innsbruck, ULB Tirol, Cod. 563). (1r) In resurrectione dni (AH 53, Nr. 36, Beginn fehlt). Inc. mut.: ] manus baptizandus et perfert fraudes temptatoris fugit persequentum lapides … (= Str. 6, V. 2). (1v) De ascensione domini (AH 53, Nr. 67). – (1v) De s. spiritu (AH 53, Nr. 70). – (2r) De sancta trinitate (AH 53, Nr. 81). – (2v) De apostolis (AH 53, Nr. 228). – (2v) De virginibus (AH 50, 164 Cod. 154 Nr. 271). – (3r) In dedicacione ecclesie (AH 53, Nr. 247). – (3v) De sancta Maria (AH 40, Nr. 119). – (3v) Item de sancta Maria (AH 54, Nr. 192). 2 (4r) Orationes de una virgine. Tit.: De una virgine non martire. – Inc.: Deus qui nos hodie beate virginis tue annua sollempnita te letificas … – Secreta. Grata tibi sit domine hec oblatio plebis tuę quam tibi offerimus in honore sanctę N virginis tuę ut cunctis proficiat ad salutem. – Postcommunio. Adiuvent nos quesumus domine et hec misteria … 3 (4r–5vb) Von späterer (Neustifter?) Hand: Officium de s. Matthaeo. Nicht mit Breviarium Romanum übereinstimmend. Mit Antiphonen, Responsorien und Versikel. Lektionen aus der Vita s. Matthaei auctore incerto nach Rom, Vaticana, Cod. Vat. lat. 1188 (Acta Sanctorum quotquot toto orbe coluntur … collegit J. Bollandus. Venetiis 1867, Sept. VI 221–223) und Mt 9,9. Ps 33, 63, 74 und 98. Inc.: Regem apostolorum dominum venite adoremus … – Expl. mut.: … Quem etiam proficien tibus eiusdem gratie crementis de communi discipulorum numero ad apostolatus gradum pro monuit nec solum predicandi verum etiam scribendi evangelium illi ministerium commisit ut qui terrestrium omiserat celestium [ 4 (6r–v) Orationes ante missam. (6r) (rot) Cum sacerdos deponit vestem dicat. (schwarz) Exue me domine veterem hominem … (Jungmann I 362 Anm. 13). (6r) Ps 84, 85, 115, 116. Pater noster. (6r) Preces (vgl. Jungmann I 355). Ps 43,26. Ps 32,22. Ps 84,7. Ps 84,8. Ps 142,2. Ps 18,13. Ps 101,2 (jeweils nur Textbeginn). (6r) Oracio. Aures tue pietatis mitissime Deus inclina precibus meis … (Jungmann I 355). (6r) Quando manus lavat. Largire sensibus nostris omnipotens Deus … (Jungmann I 361 Anm. 8. PL 78, 245). (6r) Ad humerale. Indue me domine vestimento salutis et circumda galea fortitudinis … (vgl. Jungmann I 367, dort leicht abweichend zum Anlegen der Albe). (6r) Ad albam. Indue me domine vestimento salutis et iusticię et intellectu spirituali (vgl. Jung mann I 367, leicht abweichend). (6r) Ad zonam. Precinge domine lumbos mentis meę pater clementissime et circumcide vitia cordis mei (Jungmann I 368). (6v) Ad stolam. Stola iusticię circumda domine cervicem meam … (Jungmann I 371). (6v) Ad casulam. Indue me domine ornamento humilitatis et karitatis et concede michi protectio nem … (Jungmann I 372 Anm. 66). (6v) Ad fanonem. Da domine virtutem manibus meis ad abstergendam omnem maculam … (Jungmann I 369). (6v) Letzte Formel. Fac me queso omnipotens Deus ita iusticia indui … (Jungmann I 372 Anm. 68). 5 (6v–7r) Messgebete von Stufengebet bis Opferung. 6 (7v) Messgebete zur Kommunion (Friedensgebet bis Gebete nach der Kommunion). 7 (7v–8r) Orationes post missam (wie im Missale Romanum, vgl. Jungmann II 571–572). Canticum trium puerorum (Dn 3,57–88), Ps 150, Pater noster (jeweils nur Textbeginn). Confi teantur. Sacerdotes tui. Non nobis domine non. Esto nobis domine. Domine exaudi orationem und Gebet Deus qui tribus pueris mitigasti flammas ignium concede propicius ut nos famulos tuos non exurat flamma vitiorum. Cod. 154 165 8 (8r–9v) Praefationes. Sanctus. (8r) De nativitate domini. – In epiphania domini. – (8v) In Quadragesima. – De resurrectione. – De ascensione domini. – De s. spiritu. – (9r) De s. trinitate. – De apostolis. – De s. Maria. – (9v) Praefatio communis. Anschließend Sanctus. 9 (10r–12r) Canon missae bis Agnus Dei. Bl. 10v zwei Zeilen radiert (wohl keine weitere Heiligen aufzählung) zw. Cosme et Damiani und Et omnium sanctorum tuorum quorum meritis preci busque concedas. 10 (12v–38v) Sacramentarium. 10.1 (12v–30v) Temporale. (12v) De nativitate domini. – De s. Maria infra nativitatem domini. – Dominica post nativitatem domini. – (13r) De sancto Vincentio. – De sancta Agna (!) virgine. – De sancto Mathia. – (13v) In annuntiatione s. Marie. – Dominica I. post epiphaniam. – (14r) Dominica II. – Dominica III. – Dominica IIII. – (14v) Dominica V. – Dominica in LXXa. – Dominica in LXa. – (15r) Dominica in Quinquagesima. – Feria IIII. in capite ieiunii. – Ad missam. – Feria V. – (15v) Feria VI. – Sabbato. – (16r) Dominica I. in Quadragesima. – Feria II. – Feria III. – (16v) Feria IIII. – Feria V. – (17r) Feria sexta. – Sabbato. – Dominica II. – (17v) Feria II. – Feria III. – Feria IIII. – (18r) Feria V. – Feria VI. – (18v) Sabbato. – Dominica III. – Feria II. – (19r) Feria III. – Feria IIII. – Feria V. – (19v) Feria VI. – Sabbato. – Dominica in media Quadragesima. – (20r) Feria II. – Feria III. – Feria IIII. – (20v) Feria V. – Feria VI. – (21r) Sabbato. – Dominica in passione. – Feria II. – (21v) Feria III. – Feria IIII. – (22r) Feria V. – Feria VI. – Sabbato. – (22v) (Dnca in palmis fehlt) Feria secunda. – Feria III. – (23r) Feria IIII. – (Kartage fehlen) De resurrectione domini. – (23v) Dominica in octava pasche. – Dominica II. – Dominica III. – Dominica IIII. – (24r) Dominica V. – De ascensione domini. – (24v) Dominica post ascensionem domini. – In sancto die pentecostes. – Dominica I. post pentecosten. – (25r) Dominica secunda. – Dominica III. – Dominica IIII. – Dominica V. – (25v) Dominica VI. – Dominica VII. – (26r) Dominica VIII. – Dominica VIIII. – Dominica X. – (26v) Dominica XI. – Dominica XII. – Dominica XIII. – (27r) Dominica XIIII. – Dominica XV. – Feria IIII. IIIIor temporum. – (27v) Feria VI. – S abbato. – Ad missam. – (28r) Dominica XVI. – Dominica XVII. – Dominica XVIII. – (28v) Dominica XVIIII. – Dominica XX. – Dominica XXI. – (29r) Dominica XXII. – Dominica XXIII. – Dominica in adventu domini. – (29v) Dominica II. – Dominica III. – Feria IIII. – (30r) Feria VI. – Sabbato. – (30v) Ad missam. – Proxima dominica ante nativitatem domini. 10.2 (30v–36r) Sanctorale und Missae votivae. (30v) De sancta Margareta. – (31r) De sancto Matheo apostolo. – In dedicatione ecclesie. – (31v) De sancta trinitate. – Feria II. de sapientia. – Feria III. de sancto spiritu. – (32r) Feria IIII. de angelis. – Feria V. de karitate. – Feria VI. de sancta cruce. – (32v) Sabbato de s. Maria. – De omnibus sanctis. – (33r) In basilica cuiuslibet sancti. – Pro papa vel episcopo. – (33v) Pro omni gradu ecclesie. – Pro peccatis. – Pro pluvia. – (34r) Pro serenitate. – Pro salute vivorum. – Pro tribulatione. – (34v) Pro amicis. – (35r) Pro iter agentibus. – Generalis. – (35v) Item generalis. 10.3 (36r–38v) Missae pro defunctis. (36r) In depositione defuncti. – In anniversario. – (36v) Pro episcopo (bzw. darübergeschrieben: episcopis). – Pro sacerdote (darübergeschrieben: sacerdotibus). – Pro congregatione. – (37r) Pro uno defuncto. – Pro parentibus. – (37v) Pro femina. – Pro elemosinariis. – (38r) Pro in cimite rio. – Pro nuper defunctis. – (38v) Communis oratio. 11 (38v–47v) Lectionarium (Episteln und Evangelien). Temporale und Sanctorale gemischt. (38v) De nativitate domini [in die]. – (39v) Infra nativitatem domini de s. Maria. – (40v) De s. Vincentio. – (41r) De s. Mathia. – (41v) De annuntiatione s. Marie. – (42r) De resurrectione. – (42v) De ascensione domini. – (43v) De die sancto Pentecostes. – (44r) De sancta Margareta. – 166 Cod. 155 (44v) De sancto Matheo apostolo. – (45r) In dedicatione ecclesię. – (45v) De sancta trinitate. Zwei Lesungen bzw. Evangelien. – (46v) De sancta cruce. – De sancta Maria. Mit zusätzlichem Evangelium Infra pascha (Io 19,25). – (47r) Pro peccatis. – Pro defunctis. (Ir–v) leer. W. N. Cod. 155 SAMMELHANDSCHRIFT Pap. I, 164, I Bl. 290 × 205. Umfeld Neustift (u. a. Pfalzen), 7. Jz. 15. Jh. (u. a. 1463). B: Vereinzelte Bl. stark fleckig, Bl. 111 und 112 sowie gegen Ende Wasserschaden. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichen mit Datierung übereinstimmend. Identische Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 773 (dat. 1464). Bl. 53–62 wohl Lage aus einer ca. 1425/30 angelegten Hs. (s. Wasserzeichenbefund). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + VI12 + (V–5)17 + 2.VI41 + (V+1–2)50 + (II+V)64 + 3.VI100 + (VI–1+1)112 + (VI–2)122 + (VII+2)138 + (V–5)143 + (VI–5)150 + VII164 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. der gesamten zweiten Lage herausgeschnitten. Gegenbl. zu Bl. 43, 44, 106, 113 und 114 sowie zu Bl. 139–148 heraus geschnitten (jeweils ohne Textverlust). Bl. 42 Einzelbl., auf Bl. 43 aufgeklebt. Bl. 112, 137 und 138 ein gehängte Einzelbl. Zw. Bl. 18 und 19, 84 und 85 sowie 105 und 106 jeweils ein kleines Einzelbl. eingefügt. Diese Blätter bei der Foliierung zwar berücksichtigt, doch Zahl des darauffolgenden Blattes wiederholt. Reste von alten Ledersignakeln sowie neuzeitlichen gelben Papiersignakeln der ULB Tirol. S: Bl. 1ra–50ra: Schriftraum 240/265 × 155/160. Zwei Spalten, meist von Blindlinien, vereinzelt von feinen Tintenlinien gerahmt, zu 45–50 Zeilen auf Blindlinierung. Bastarda von einer Hand mit unterschiedlichem Duktus. Bl. 51r–105v: Schriftraum 230/245 × 140/150, von Blindlinien (Bl. 53r–62v: von feinen Tintenlinien) gerahmt, zu 34–41, meist 37 Zeilen auf Blindlinierung. Bastarda von zwei Händen: 1) Bl. 51r–52r und 63r– 105v sowie eingehängtes Einzelbl. 106r–v mit breitem und rundem Duktus, dat. 1463 (Bl. 105r). Schreiber: Matheus Tegedener, vgl. Krämer, Scriptores; 2) Bl. 53r–62v. Bl. 106ara–122ra: Schriftraum 250/255 × 150/155. Zwei Spalten, von Blindlinien gerahmt, zu 47–50 Zeilen auf Blindlinierung. Bastarda. Bl. 123r–140r: Schriftraum 240/255 × 145/155, von Blindlinien gerahmt, zu 47–56 Zeilen auf Blind linierung. Bastarda mit engem und kleinem Duktus. Bl. 141ra–150ra: Schriftraum 240/250 × 150/160. Zwei Spalten, von Blindlinien gerahmt, zu 46–53 Zeilen. Bastarda. Bl. 151r–163v: Schriftraum 245/255 × 80/95, von Blindlinien gerahmt, zu 28–37 Zeilen. Gleichmäßige Bastarda. Bl. 151r–155v am rechten Textrand Kommentar von gleicher Hand. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unter- und Durchstreichungen, Rahmungen sowie tw. Rand notizen. Zwei- bis fünfzeilige rote Lombarden, z. T. mit Punktverdickungen. Rubrizierung tw. nur sehr spärlich. Bl. 48vb–50ra, 72v–100v, 101v–131r, 132r–134v Rubrizierung nicht ausgeführt (tw. Platz für Initialen und Paragraphzeichen freigelassen). E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck IUS CANON. in Gold rahmung, im dritten Signaturschild (s. bei G). Kapitale mit blauem und naturfarbenem Zwirn umstochen, freiliegend. Schnitt rot gesprenkelt. Spiegel und Vor- bzw. Nachsatzbl. Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G). Spiegel des HD leer. G: Bl. 164v Besitzvermerk 15. Jh. Iste liber est monasterii sancte Marie vulgariter ad Novamcellam. Am Spie gel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II (Tinte) 2 K 3 (Bleistift), Nr. 155 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 155. Bl. Ir Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain Cod. 155 167 an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe über lassen. Bl. Ir Stempel der Soprintendenza Tridentina 20. Jh. L: 1 2 Wilhelm II 109. – Wretschko–Sprung 17. – H. Caplan, Mediaeval Artes praedicandi. A Hand-List und A Supplementary Hand-List (Cornell Studies in classical philology 24 und 25). Ithaca, N. Y. 1934 und 1936, Nr. 14. – Th.-M. Charland, Artes praedicandi. Contribution à l’histoire de la rhéthorique au moyen-âge (Publications de l’Institut d’études médiévales d’Ottawa 7). Paris, Ottawa 1936, 44. – Kristeller I 439. – Dolezalek II o. S. – Colophons IV Nr. 13447. – Furtenbach–Kalb 21. – Neuhauser, Neustift 89. – S. Furten bach, Ars notariatus. Ein kurialer Notariatstraktat des 15. Jahrhunderts. Österreichisches Archiv für Kirchen recht 30 (1979) 3–22, 299–327. – Katalog ULB Tirol II 133, 205–206. – Katalog ULB Tirol II, Beiheft, Taf. 69. – Stefani 102–103. – Peintner, Stiftsbibliothek 122. – Krämer, Scriptores, s. v. „Tegedener, Matheus“. (Ir–v) leer. (1ra–6va) Franciscus de Zabarellis: De horis canonicis (Ed.: V. O. Ludwig, Ein neuaufgefunde ner Traktat Franz Zabarellas [Jahrbuch des Stiftes Klosterneuburg III]. Wien, Leipzig 1910, 151–178). Inc.: Repetiturus decretalem [X.3.41.1.] presbyter que prima sub rubrica de ce[lebratione] mis sarum situatur et ponitur … – Expl.: … in c. Cum Marthe Ibi materia plenius terminatur scilicet de temp[ore] ordi[nis] et sic est finis. Vgl. Schulte GQ II 283–285. (6va) Direkt anschließend Schlussbemerkungen zum Traktat des Franciscus de Zabarellis (nicht in Edition enthalten): Sequitur prima conclusio. Quamvis beneficiatus per alias horas canonicas … Item decretalem presbiter repetiit dominus Franciscus de Saberellis (am Blattrand korr.: Sarabellis) qui post ea fuit creatus in cardinalem et tunc fuit nominatus Florentius qui obiit in concilio Constantiensi. (6va–15vb) Sammlung kanonistischer Texte (Verweise u. a. auf Iohannes Calderinus, Raimun dus de Pennaforte). (6va) Inc.: Numquid Iudeus litigans cum Christiano iurabit de calumpnia super ewangelio vide tur … (6vb) Tit.: De adulterio et stupro. – Inc.: An maritus uxori adultere subtrahere possit debitum cohabitacionum et dotem sine iudicio ecclesie. Tua fraternitas hic tractantur iura an vir uxori adultere … (7ra) Tit.: De pe[nitentia?] et re[missione?]. – Inc.: Tutela bona confessionis quando confitetur mulier se habere filium spurium inter legitimos. Officii mulier que partum supponit alienum vel de adulterio concepit … (7rb) Tit.: De uno filio inter multos alios legitimos. – Inc.: De quadam muliere dicebatur una vice casus supradictus fuisse … (7va) Inc.: An reddituarii teneantur propriis sumptibus deferre dominis redditus et presertim ecclesiarum. Et videtur quod sic quia redditus ecclesie videntur esse dos ipsius … (7vb) Tit.: Casus diversi et periculosi in confessionem contingentes. – Inc.: Si uxor alicuius viri quia non potest prolem habere timens quod possessio viri ad alios devolveretur … (8rb) Inc.: Aliquis fuit testis in causa matrimoniali qui falsum protulit testimonium vel aliqua dixit de causa matrimoniali se scire que in veritate ignoravit … (8va) Inc.: Aliquis vovebat perpetuam continenciam postmodum non obstante voto … (8va) Inc.: Aliquis habens uxorem postmodum commisit cum filia uxoris sue adulterium … (8va) Inc.: Aliquis cognovit matrem et filiam vel duas sorores vel duas consaguineas mulieres … (8vb) Inc.: Aliquis contraxit sponsalia cum aliqua muliere et contracto matrimonio cognovit carnaliter mulierem sponse ante sponsam … (8vb) Inc.: Aliquis dicit confessori suo quod mulierem quam nunc habet duxit in uxorem in facie ecclesie … (9ra) Inc.: Aliquis segetem meam sevit tempore oportuno animo surripiendi sed nichil de ea asportavit … 168 Cod. 155 (9ra) Inc.: Aliquis venit ad confessorem et dicit ei peccata sua et dicit se non posse abstinere a luxuria … (9ra) Inc.: Aliquis furiosus interfecit hominem postea factus sane mentis se hominem interfecisse confitetur … (9rb) Inc.: Aliquis confitetur aliquod crimen sacerdoti et episcopus excommunicat omnes huius criminis consocios … (9rb) Inc.: Aliquis est surdus et mutus vel ita eger quod non potest peccata sua confiteri sacerdoti … (9rb) Inc.: Aliquis peregrinus vel viator seu negociator vel mercator venit ad extraneum sacerdotem … (9rb) Inc.: Aliquis confitetur peccata sua sacerdoti et dicit quod non potest cessare … (9va) Inc.: Si quis revelet peccatum suum sacerdoti non ut peniteat sed ut consilium et auxilium ab eo recipiat … (9va) Inc.: Aliquis in infirmitate petit sacerdotem et antequam ad eum venerit sacerdos … (9va) Inc.: Aliquis habetur pro prelato et in veritate non est prelatus audit confessiones parro chianorum illius parrochie … (9vb) Inc.: Mulier uxor alicuius confitetur sacerdoti se tenuisse filium proprium coram episcopo … (9vb) Inc.: Filius carnalis dicit confessori suo se duxisse in uxorem illam quam pater suus bap tizaverit … (10ra) Inc.: Iohannes qui non est pater meus nec consanguineus duxit matrem meam in uxorem suam. Ipsa moritur postmodum … (10ra) Inc.: Est dubitacio de monialibus occulte corruptis quia in quibusdam monasteriis consue tudo est quod sole virgines … (10rb) Tit.: De sacramento eukaristie. – Inc.: Nemo potest conficere corpus Christi nisi sacerdos qui rite fuerit ordinatus secundum claves ecclesie … (13ra) Tit.: De casibus diversis et periculosis in missa contingentibus. – Inc.: Presbiter potest supplere missam ab episcopo inchoatam si episcopus non poterit propter infirmitatem … (14rb) Inc.: Misse sunt celebrande in locis Deo consecratis vel in locis in quibus episcopus permiserit celebrare quia melius est missam non celebrare aut non audire … (14va) Inc.: Utrum habens confessionale a se[de] ap[ostolica] secundum formam consuetam possit sibi racione istius eligere in confessorem quemcumque sacerdotem non suspensum … (14vb) Inc.: Utrum racione confessionalis quod dari consuevit possit quis absolvi in casibus episcopalibus a quolibet sacerdote … (14vb) Inc.: Utrum sacerdos in confessione eam absolvere posset cum qua peccavit fornicarie … (15ra) Inc.: Quid agendum sit dum quis habeat aliqua peccata a quibus ille cui confiteatur non potest absolvere. Dicit beatus Tho[mas] … 3 (15ra) Kanonistische Notiz zur Eucharistie. Tit.: Nota. – Inc.: I. Cor. XIIII (19) Sed eciam volo quinque verba sensu meo loqui ut et alios instruam … Nota vir debet se abstinere ab uxore sua ante sacrum et communionem per III vel IIII vel VI dies … (15v) leer. 4 (16ra–vb) ‹Ps.-Augustinus›: Sermo 277: De reddendis decimis = ‹Caesarius Arelatensis›: Ser mo 33 (PL 39, 2266–2268. CC 103, 143–146. – CPL Nr. 284). Tit.: Sermo de reddendis decimis. – Expl. (gegenüber PL und CC gekürzt): … in illa sacrilega lavacione moriantur vel mortem corporis timeant qui de anime sue salute non curant. 5 (16vb–17va) Kurztraktat zu veri und falsi praedicatores. Inc.: Veri predicatores precipue quibus non sunt certe elemosine assignate libencius vadunt ad eos convertendos qui nondum ad fidem Christi conversi sunt quam ad illos qui iam conversi Cod. 155 169 sunt … – Expl.: … ducit eos ad heresim non enim simplices sed studiosi ad here[sim] confluxe runt. Ty. (?) VI. Darstellung der 50 Zeichen zur Identifikation von Ps.-Predigern ab Bl. 17va übereinstimmend mit der Tabula de signis per quae pseudo-praedicatores discerni possunt a veris zu Guilelmus de Sancto Amore: Collationes catholicae et canonicae scripturae 4 (Druck: Opera omnia. Konstanz 1632, 487–490). (17va–b) Unmittelbar anschließend Tabelle mit Auflistung der verschiedenen Ordensgemeinschaften. Tit.: Ffratres mendicantes et cappam habentes et quasi omnes regulas eorum deserentes. – Inc.: Ordo fratrum Predicatorum. Ordo fratrum Minorum … – Expl.: … Ordo Celestinorum. In via Dei alius sic alius sic ibat sed utrum hec predicta debite fuerit et plene dubitatur a quam p luribus. Gliederung nach: Monachi. – Regulares canonici. – Ffratres degentes sub regula. 6 (18ra–45rb) Begriffserklärungen zu den Canones. Innerhalb eines Buchstabens alphabetische Anordnung, Alphabet jedoch nicht konsequent eingehalten. Wohl Kompilation aus mehreren Rechtslexika. (18ra) Prolog. Inc.: In nomine domini Amen. Quia in libris sacrorum Canonum et in legibus imperialibus plerumque restat difficultas vocabulorum … – Expl.: … per superius dictum quia omnium habere memoriam etc. (18ra) Vocabularius. Text. Inc.: Abigeus dicitur ille qui pecora alicuius a pascuis et armentis subducit … Abolicio est ab accusando cessacio … – Expl.: … Zenodochius dicitur qui zenonem alicuius civitatis procurat … Zenodochium dicitur locus hospitum vel domus peregrinantibus … de sanctissimis episcopis coll. IX post principium etc. Lit.: E. Seckel, Beiträge zur Geschichte beider Rechte im Mittelalter I. Tübingen 1898, 323–335, Teiledition 358–365. Parallelüberlieferung: u. a. München, BSB, Clm 3563, Bl. 246r. 7 (45va–48ra) Termini librorum I–V Decretalium. Tit.: Exposiciones terminorum I li. Decretalium. – Inc.: Constitucio est ius relatum in scriptis re dactum. Vel aliter est ius ubi non sequitur pena. Sed sanxio est ius ubi sequitur pena. Consultacio est in qua … – Expl.: … Celebracio missarum est cum missa propter vivos et mortuos cantetur. Regula dicitur virga … est multorum constructibilium colleccio. (45va) Lib. 1. – (46ra) Lib. 2. – (46va) Lib. 3. – (47ra) Lib. 4. – (47rb) Lib. 5. Parallelüberlieferung: u. a. Melk, Stiftsbibl., Cod. 1894, Bl. 181r. 8 (48ra–49va) Glossar, Verzeichnis verschiedener Autoren juristischer Werke, Abkürzungsver zeichnis. (48ra) De nominibus doctorum et primo quoad doctores sacre theologie qualiter iuvenes debent legere. (48va) Doctores qui super decreto scripserunt sunt hii. (48va) Hii doctores scripserunt ad Decretales. (48vb) Abkürzungsverzeichnis in alphabetischer Reihenfolge. 9 (49vb–50ra) Argumenta quibus valet arguere in canonico iure. Tit.: Nota argumenta quibus valet arguere in canonico iure. – Inc.: Primo reprobatur superfluitas quia superflua sunt resecanda ab arte id est a iure canonico … – Expl.: … de pactis pactus. Quod semel est dictum manet irrevocabile verbum de proba per tuas. Tit. und Inc. wie Augsburg, UB, Cod. II.1.4o 21, Bl. 146v. (50rb–vb) leer. 170 Cod. 155 10 (51ra–105r) Iohannes de Auerbach: Processus iudiciarius (Ed. Th. Muther. Halae 1873, 3–322). Schlussschrift mit Datierung und Schreibernennung: Terminatus est hic iudiciarius processus Hin rici Aurbach decretorum doctoris per Matheum Tegedener de Iuterbock presbiterum Brandebur gensis diocesis. In domo et habitacione reverendi patris Leonardi plebani in Phalczen (gestrichen: anno) Briccinensis diocesis anno domini MCCCCLXIII in octava corporis Cristi (16. Juni 1463). Lit.: Schulte GQ II 301–302; H. Boockmann, Aus den Handakten des Kanonisten Johannes Urbach (Auerbach). Deutsches Archiv für Erforschung des Mittelalters 28 (1972) 497–532. 11 (105v–106v) Declaratio arboris consanguinitatis (vgl. CIC, ed. Friedberg I 1429–1430). (105v) § 16–17 In linea equali colleteralium (!) quoto gradu quis distat a communi stipite toto distant inter se … (106r) Inc.: Aut fratres sunt ex utroque coniuncti aut non. Si ex utroque coniuncti et tunc aut soli fratres extant … – Expl.: … autem de here[ditate] ab intestato. (106v) Inc.: Nota. Detur quid tunc si superest filius fratris ex uno parente coniuncti … – Expl.: … filiis fratris utriusque coniunctis. Idem est eciam dicendum de nepotibus. 12 (106ara–112v) Exzerpte aus ‹Bernardus Parmensis›: Casus longi Decretalium (Druck: GW 4092–4105). (106ara) Prolog. Tit.: Prologus. – Inc.: [R]ex pacificus premissa salutacione sic pone casum in ista constitucione. Rex pacificus id est dominus noster Ihesus Christus … – Expl.: … sine licencia sedis apostolice speciali. (106ara) Text. Tit.: De summa trinitate et fide katholica firmiter credimus. – Inc.: Nota quod post simbolum apostolorum quod dicitur Credo in Deum concilia predicta fuerunt olym a sanctis patribus de summa trinitate et fide katholica contra quosdam hereticos de quibus quattuor con ciliis legitur … – Expl.: … inferiores prelati sine diocesani consensu in ecclesiis suis non possint instituere clericos peregrinos. (113r–114v) leer. 13 (115ra–118ra) Ars notariatus (Druck: u. a. GW 2648. Ed. Furtenbach 308–327). Lit.: R. Stintzing, Geschichte der populären Literatur römisch-katholischen Rechts. Leipzig 1867, 297–299; L. v. Rockinger, Ueber Formelbücher vom dreizehnten bis zum sechzehnten Jahrhundert als rechtsgeschichtliche Quellen. München 1855, 66 Anm. 132; Kristeller I 439; Dolezalek II o. S.; Furtenbach 3–22, 299–327. 14 (118rb–122ra) ‹Ps.-Iohannes Andreae›: Ordo iudiciarius (Druck: u. a. GW M47346. Ed.: O. Riedner, Die geistlichen Gerichtshöfe zu Speier im Mittelalter II. Paderborn 1915, 5–48. – Hamesse Nr. 3001–3002). Anschließend (121vb): Iura hic non allegantur sed pocius obmittantur non propter ignoranciam doctorum sed propter imbecillitatem discencium ad quorum profectum hec summula conscripta est … sed perfectum illorum quos informat. Lit.: Schulte GQ II 225–226. Vgl. auch R. Stintzing, Geschichte der populären Literatur des römisch-kanonischen Rechts. Leipzig 1867, 202–220. Zur Verfasserfrage s. L. v. Rockinger, Über einen ordo iudiciarius, bisher dem Johannes Andreae zugeschrieben. München 1855; L. Fowler-Magerl, Ordines iudiciarii and libelli de ordine iudi ciorum (Typologie des sources de moyen âge occidental 63). Turnhout 1994, 98. Anschließend (122ra): Inc.: Est discrecio testantisque suprema voluntas accio … – Expl.: … favor odia dolus. (122rb–vb) leer. 15 (123r–138r) ‹Ps.-›Albertus Magnus: Secreta mulierum, mit Kommentar (Druck: GW 719– 759a. – Thorndike–Kibre 635). Verglichen mit GW 724, leicht abweichend. Cod. 155 171 (123r) Kommentarprolog. Inc.: [H]omo est optimum eorum que sunt in mundo IIII Ethicorum et mundus … – Expl.: … et non pure naturalis sed partim medicinalis et partim naturalis. (123r–138r) Text. Tit.: Titulus: Incipit Secreta mulierum et virorum ab Alberto Magno compo sita. – Inc.: [D]ilecto sibi in Christo socio et amico N. clerico de tali loco vere sapiencie et augmentum continuum vite presentis. Iste liber cuius subiectum est ens mobile contractum ad naturam secretorum mulierum … – Expl.: … Hiis visis dictis finem imponamus … per infinita secula seculorum Amen. (138r) Unmittelbar anschließend. Inc.: Nota. Coitus maxime exsiccat quia per ipsum recedit magna humiditas … – Expl.: … longitudo et breviacio vite in hominibus est a natura et eius constellacione et illud eciam maxime habet variari etc. (138v) leer. Lit.: L. Thorndike, A History of Magic and Experimental Science II. London 1923, 739–745, 749–750; M. Schleissner, „Secreta mulierum“. Verfasserlexikon2 VIII 986–993. 16 (139r–140r) Ars praedicandi (Caplan Nr. 14. Charland 44 mit Zuschreibung an Henricus de Lan genstein, vgl. Caplan 7. Th. Hohmann, Initienregister der Werke Heinrichs von Langenstein. Traditio 32 (1976) 399–426, hier Nr. 16). Inc.: Ars predicandi est sciencia docens de aliquovis termino vel ente aliquid dicere … – Expl.: … Vicia sermocionantis (!) Ignorancia predicantis Infacundia … Digressio remota Applicacio defectuosa. (140v) leer. 17 (141r–143v) ‹Iohannes Marchesinus›: Super Psalterio, aus: Mammotrectus super Bibliam (Druck: GW M20819. – Stegmüller RB Nr. 4776). Gegenüber Druck tw. stark gekürzt. Inc.: Beatus vir. Pestilencie contagie (!) quod dum unum apprehendit celeriter transit ad plures quia velut incendium … Domine quid multiplicati. Soporatus id est sopitus … Verba mea. Patens id est apertum decidant id est cadant … Domine ne in furore. Stratum id est lectum … Domine Deus meus. Decidam id est cadam … – Expl.: … Laudate dominum de celis. … Id est in metallis permixtis acutum sonum facientibus. 18 (144ra–150ra) Auctoritates philosophorum. Tit.: Auctoritates philosophorum. – Inc.: Tales dicit [A]ntiquissimum encium est Deus ingenitum et in evum. Optimum encium mundus nam opus Dei … Solon dicit Quid est verbum Imago ope rum quo firmatur verbum … Pitacus dicit Optimas esse victorias que sine sanguinis effusione fiant. Item prudentium virorum esse priusquam fiant difficilia … – Expl.: … Secundus philo sophus dicit Athenis claruit tempore Adriani imperatoris qui philosophatus est omni tempore silentium habere … fortuna sine sollicitudine felicitas etc. Explicit materia philosophorum sed non omnium. Thales – Solon – Chilo – Pittacus – Bias – Cleobolus – Periander – Pythagoras – Simonides – Isocrates – Crispus – Socrates – Democritus – Hippocrates – Heraclitus – Diogenes – Plato – Aristoteles – Epicurus – Theophrastus – Hecaton – Cicero – Cato – Plautus – Iulius Caesar – Terentius – Varro – Existus Xistus (Sixtus?) – Athenodorus – Valerius – Quintilianus – Plinius – Ptolomaeus – Galenus – Symachus patricius – Secundus philosophus. Dasselbe Inc. sowie ähnliche Auswahl an Autoren in St. Gallen, Stiftsbibl., Cod. 482, Bl. 362r. Vgl. tw. Diogenes Laertius, De vita et moribus philosophorum libri decem. Coloniae Agrippinae 1535. (150rb–vb) leer. 19 (151r–155r) Theobaldus: Physiologus cum commento (Ed.: P. T. Eden, Theobaldi „Physiolo gus“ [Mittellateinische Studien und Texte 6]. Leiden, Köln 1972, 24–72. – Thorndike–Kibre 1584). 172 Cod. 156 Kommentar, vorwiegend rechts des Textes. Inc.: Iste liber Phisiologus intitulatur et dicitur a physis id est natura … – Expl.: … Allegoria secunda pili elephantis id est doctrina Christi que si incenduntur in aliqua domo id est homine recedunt omnia animalia venenosa. 20 (155v–163v) Avianus: Fabeln (Nr. 1–28). Mit Kommentar (nur Bl. 155v) (Druck: GW 3110. Ed. K. Lachmann. Berolini 1845). Kommentar nicht im Druck enthalten. Inc.: [R]ustica deflenti puero iuraverat olym / Ni taceat rapido quod foret esca lupo … – Expl.: … Qua fieri posset cum racione nocens / Vix castigator cui semita recta negatur / Que mala mens didicit perdere vix poterit. Kommentar (zu Fabeln Nr. 1 und 2). Inc.: Liber Aviani poete in quo primo introducit fabulam de villana et lupo ostendens in eo quod fraudulenti mulieri non est fides … – Expl.: … dabitur et incurrit periculum. (164r–v) leer. (Ir–v) leer. P. A. / A. P. Cod. 156 HUGO DE SANCTO CARO Perg. I, 88, I* Bl. 290 × 210. Nordfrankreich (Raum Paris), um 1300. B: Perg. unregelmäßig beschnitten, bes. am Beginn stark verschmutzt, tw. löchrig, Nahtspuren. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 4.V40 + 2.IV56 + 2.VI80 +IV88 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Nach Bl. 88 fehlt mindestens eine Lage. Am Lagenanfang Kustoden in arabischen Ziffern, am Lagenende in roten bzw. schwarzen römischen Zah len. Am oberen rechten Blattrand spätmittelalterliche Foliierung in arabischen Ziffern. S: Schriftraum 200/205 × 135/140, von seitlich zweifachen Tintenlinien gerahmt, zu 46 (Bl. 1ra–10vb) bzw. 40 Zeilen (Bl. 11ra–88vb) auf Tintenlinierung. Bl. 1r–20v am äußeren Blattrand dritte gerahmte Spalte, tw. liniert. Zirkelstiche. Littera Parisiensis um 1300. Notizen, Nachträge, Auctoritatesangaben, gerahmte Inhaltshinweise von verschiedenen Händen. A: Rote Auszeichnungsstriche, Durchstreichungen, Überschriften, Rahmungen, Notizen, zweizeilige (Bl. 1ra und 2rb dreizeilige) Lombarden mit kopfstempelförmiger Aussparung und einfachem Dekor. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Feld Golddruck HUGO IN PROVER. in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen, Bleistiftnotiz (s. bei G). Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 4 (Tinte, Bleistift), Nr. 156 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 156. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kaeppeli Nr. 1989. – Stegmüller RB Nr. 3677 Suppl. – Steer 36. – Katalog ULB Tirol II 133, 206. (Ir–v) leer. (1r–88v) Hugo de Sancto Caro = ‹Ps.-Nicolaus de Gorran›: Postilla in Proverbia, unvollständig: cap. 1–25 (Druck: Opera omnia III. Venetiis 1754. – Stegmüller RB Nr. 3677 und Nr. 3677 Suppl., mit Nennung der Hs. – Kaeppeli Nr. 1989, mit Nennung der Hs.). Cod. 158 173 Tit.: Incipiunt notule compilate a magistro Hugone fratre ordinis Predicatorum super Proverbia. – Inc.: Dicit Ecclesiasticus XXX IX b (!) occulta Proverbiorum exquirit sapiens et in absconditis parabola rum … – Expl. mut. (cap. 25): … invito auditore libenter refert ut dicit Ieronimus. Et ubi (!) ne esset auditor ne esset detractor. Unde super XXIIII [ (Ir–v) leer. P. K. Cod. 158 AUGUSTINUS Pap. III, 69, I Bl. 285 × 205. Süddeutscher Raum, um 1400. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: Neustift, Stiftsbibl., Cod. 83 (dat. 1400) und Cod. 183. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + (VI+1)III,11 + 2.VI35 + V45 + 2.VI69 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. II eingefügtes Einzelbl., an Bl. III angeklebt. Am Lagenende Kustoden in arabischen Ziffern. Ab Textbeginn am oberen Blattrand Foliierung 15. Jh. 1–69, Hinweis auf ehem. Sammelhandschrift (s. bei G). S: Schriftraum 220/225 × 150. Zwei Spalten, von Tintenlinien gerahmt, zu 54–57 Zeilen. Bastarda (= Cod. 83, Schreiber 1). A: Rote Auszeichnungsstriche, Unterstreichungen, Über- und Schlussschriften. Flüchtige Zeigehände. Fünfbis siebenzeilige rote bzw. rot-schwarz kopfstempelförmig gespaltene Lombarden, tw. mit Punktverdickun gen und Ausläufern. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. CONFE. S. AUGU. in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. IIr zwei Besitzvermerke 15. Jh. Iste liber est monasterii s. Marie ad gracias alias ad Novamcellam dicti und Novacelle sowie Neustifter Wappen in Federzeichnung. Nach dem Inhaltsverzeichnis in dieser Hs. auf Bl. Ir gehört Cod. 158 zu einer Gruppe von urspr. tw. selb ständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammel handschrift (wie dies auch bei anderen Handschriften und Inkunabeln gemacht wurde) in sieben Einzelhand schriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. Bl. IIIr Besitzvermerk 18. Jh. Collegii Neocell. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II (Tinte) 2 H 6 (Bleistift) und Nr. 158 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 158. Bl. IIr Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Ger main an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. IIr Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 133, 206. – Peintner, Stiftsbibliothek 121. 174 Cod. 160 (Ir–v) leer. (IIr) Besitzvermerke und Stempel (s. bei G). Federproben u. a.: 1) Raymundus. Interpretacio huius nominis phaf ponit. p pater h honoris a altissimi f filius. 2) Interpretatio huius tituli (?) monachus. (tw. gestrichen:) M manducans o omnia n nichil a amans c culum h habens u vulvam s supponens (?). 3) ‹Serlon de Wilton›: Versus (Walther, Initia Nr. 10349. J. Öberg, Poèmes latins [Acta Universitatis Stockholmiensis. Studia Latina Stockholmiensia 14]. Stockholm 1965, 121, Nr. 78). Tit.: Nota si vis. – Text (Hexameter): Linquo coax ranis cra corvis vanaque vanis / ad loycam (!) pergo que mortis non timet ergo. – Hec ille. Inhaltsverzeichnis des Sammelbandes mit Blattangabe. Tit.: Contenta in hoc libro hic signata. Libri tredecim Confessionum beati Augustini episcopi 1. – Quatuor libri Dyalogorum beati Gregorii pape 77. – Tractatus De contractibus m. Heinrici de Hassia in sudore wltus tui 130. – Epistola eiusdem de ymagine mundi et eius contemptu venerando domino 160. – Speculum peccatoris. Quoniam karissi mi etc. 122 (!). – Horologium sapiencie. Sentite de domino in bonitate 166. – Tractatus de indulgenciis. Cum non paucorum 204. – Omelia Origenis super ewangelio. Maria stabat ad monumentum 209. – Epistola Eusebii de abitu beati Ieronimi presbiteri. Patri reverendissimo 216. – Sermones quidam de dedicacione ecclesie primus venit enim filius hominis 232. (IIv) leer. (IIIra) Federprobe: Beginn von Augustinus, Confessiones, Lib. 1 (PL 32, 659–661, leicht abweichend): [M]agnus dominus et laudabilis nimis. [M]agnus dominus et laudabilis nimis. Magna virtus tua … (IIIrb) Tit.: Tredecim libri Confessionum sancti Augustini beatissimi patris nostri ad proficiendum uti lissimi. (IIIv) leer. (1ra–60ra) Augustinus: Confessiones (PL 32, 659–868. CSEL 33, 1–388. – CPL Nr. 251). (1ra) Lib. 1. – (4va) Lib. 2. – (7va) Lib. 3. – (9rb) Lib. 4. – (12vb) Lib. 5. – (16ra) Lib. 6. – (20rb) Lib. 7. – (24va) Lib. 8. – (28vb) Lib. 9. Bl. 33v leer mit Vermerk Hic nullus est defectus. Ohne Textverlust. – (34ra) Lib. 10. – (43ra) Lib. 11. – (48rb) Lib. 12. – (53va) Lib. 13. (60ra) Schlussschrift: Deo gracias Amen. Explicit liber Confessionum beati Augustini primo anno etc. (60rb–69vb) nur Rahmung und Foliierung. (Ir–v) leer. U. S. Cod. 160 LAURENTIUS RUSIUS Pap. I, 55, I Bl. 290 × 210. Süddeutscher Raum, Mitte 15. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Innsbruck, ULB Tirol, Cod. 624, T. II (dat. 1449). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + (VI+1)13 + 3.VI49 + 655 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. 12 sowie 50–55 eingehängte Einzelbl. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 225 × 145, von Tintenlinien begrenzt, zu 34–38 Zeilen (Bl. 2ra–4rb auf Tintenlinierung). Bastar da. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Kapitelnennung im Register und Über schriften. Zweizeilige (Bl. 4ra vierzeilige) rote Lombarden, tw. mit Punktverdickungen, tw. mit einfachen Fadenausläufern. Bl. 1r zehn Einzelzeichnungen in Schwarz (monogrammartig) auf Blindlinierung, wohl von einer Hand. 175 Cod. 160 E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten bis vierten Feld Golddruck RUZZI MARESCHALCIA EQUORUM in Goldrahmung, im fünften Signaturschild (s. bei G). Schnitt rot g esprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 8 (Tinte, Bleistift), Nr. 160 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 160. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller VI 122. – Katalog ULB Tirol II 133, 206. – Peintner, Stiftsbibliothek 122. 1 (Ir–v) leer. (1r) Zehn Zeichnungen in Schwarz (s. bei A). 2 (1r) Rezepte zur Heilung von Pferdekrankheiten. Contra debilitatem oculorum equorum et macularum … Contra debilitatem capitis naturalem … Contra inflacionem colli equorum … Contra vermes in intestivis equorum … Quando equus patitur in pedibus defectum … Rappigen rossen. Quando equus patitur fissuras contra ungulam wlgariter in den fisselen … Quando dorsum equi leditur per sellam … Contra scabiem equorum … (1v) leer. 3 (2ra–55va) Laurentius Rusius: Marescalcia (Druck: GW M39220. – Thorndike–Kibre 766. L. Schnier, Die Pferdeheilkunde des Laurentius Rusius, Diss. Berlin 1937. W. Perino, Die Pferde arzneibücher des ausgehenden Mittelalters und der beginnenden Neuzeit, Diss. München 1957, 12–14). (2ra) Register. Tit.: Incipit tabula sive registrum. (3vb) leer. (4ra) Prolog. Tit.: Epistola sive prohemium. – Inc.: Reverendo in Christo patri et domino suo do mino N. Dei gracia sancti Adriani dyaconi cardinalis Laurentius dictus Kytius (!) mereschaltus (!) de Urbe … Inter cetera animalia que ad dignitatis humane solacium … (4rb) Text. Tit.: De natura equi. Rubrica. 2m.. – Inc.: Equs (!) calide nature iudicatur sed tempera te calor ostenditur per levitatem … – Expl.: … et mitte eum ad modum suppositorii in equi sexum aut anum. Et sic est finis Deo gratias. (55va) Schlussschrift: Explicit liber mareschalcie equorum compositus (gestrichen: per Paulum K. de Monaco et sic est finis) per Laurencium dictum Ruzzum de Urbie mareschalcum et familia rem reverendi patris et domini Neapoleonis de Ursinis sancti Adriani diaconi cardinalis. (55vb) leer. (Ir–v) leer. C. S. 176 Cod. 161 Cod. 161 IOHANNES DE DAMBACH. ORIGENES. EXPOSITIO ORATIONIS DOMINICAE Pap. I, 12, I Bl. 285 × 205. Süddeutscher Raum, Ende 14. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Innsbruck, ULB Tirol, Cod. 608, T. II. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + VI12 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Erstes und letztes Blatt des Sexternio jeweils am Vor- bzw. Nachsatzbl. angeklebt. Am oberen Blattrand Foliierung 15. Jh. 204–215, Hinweis auf ehem. Sammelhandschrift (s. bei G). Daneben Foliierung 19. Jh. durch die ULB Tirol 1–12 (hier berücksichtigt). S: Schriftraum 225 × 150/155. Zwei Spalten, von Tintenlinien gerahmt, zu 43–52 Zeilen. Bastarda. A: Bl. 1ra–8rb rote Auszeichnungsstriche und Unterstreichungen sowie zwei- und dreizeilige rote Lombarden. Bl. 6ra rot-tintenfarbener I-Versal mit kopfstempelförmiger Aussparung, Punktverdickung und Ausläufern. Ab Bl. 8va Rubrizierung nicht ausgeführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S., im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Neustift, Stiftsbibl., Cod. 158, Bl. Ir gehört Cod. 161 zu einer Gruppe von urspr. tw. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzel handschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 2 H 9 (Tinte, Bleistift) und N. 161/1 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 161. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller I 439. – Kaeppeli Nr. 2262. – J. P. McCall, Chaucer and the Pseudo Origen „De Maria Magdalena“: A Preliminary Study. Speculum 46 (1971) 491–509, hier 507. – Katalog ULB Tirol II 133, 206. 1 2 (Ir–v) leer. (1ra–6ra) ‹Iohannes de Dambach›: Tractatus de indulgentiis (Kaeppeli Nr. 2262, mit Nennung der Hs.). Inc.: Cum non paucorum erga beneficia indulgenciarum mens errabunde vagetur … – Expl.: … hoc eciam est de intencione beati Thome de Aquino in scripturis super quartum Sentenciarum etc. Sic est finis huius tractatus. Zu Iohannes de Dambach s. u. a. F. J. Worstbrock, „Johannes von Dambach (Tambach, de Tam baco, Than-, Cam-, Zam-, -bico, -buco) OP“. Verfasserlexikon2 IV 571–577. (6ra–8rb) ‹Ps.-›Origenes: Homilia de s. Maria Magdalena (Druck: J. Merlin, Origenis Adamantii Cod. 162 177 Opera III. Parisiis 1512, 129ra–131vb. – CCCPG I Nr. 1523). Tit. (am oberen Blattrand): Omelia Origenis super ewangelium Maria stabat ad monumentum. – Inc.: In illo tempore stabat Maria ad monumentum foris plorans et reliqua (Io 20,11). Omelya Origenis. De presenti solempnitate locuturus auribus vestre caritatis dilectissimi amor venit ad memoriam … – Expl.: … quia vidi dominum et hoc dixit michi cui est honor et gloria cum patre et spiritu sancto in secula seculorum Amen. Explicit omelia Origenis super ewangelium Iohannis Stabat ad monumentum. 3 3.1 (8va–9va) Expositio orationis dominicae. (8va) Inc.: [S]anctus Gregorius in prologo oracionis dominice dicente Preceptis salutaribus moniti et divina institucione formati audemus dicere … – Expl.: … eis infirmatus est tamquam dominus servis. Nos igitur per sanguinem Christi filii adopcionis effecti audemus dicere Pater noster. 3.2 (9ra) Inc.: [H]oc opusculum dividitur in tres partes scilicet in exordium in quo benivolencia captatur tractatum … Prima pars tangitur ibi Pater noster 2a ibi sanctificetur 3a in fine cum concluditur Amen. Circa primam partem est sciendum primo quod per istam partem exordialem datur nobis petendi confidencia … – Expl.: … et filii dilectissimi patrem in trepidacione appel lare debemus quid delictum est autem que offensa quam presens. (9vb–12vb) nur Rahmung und Foliierung. (Ir–v) leer. P. A. / U. S. Cod. 162 SERMONES IN DEDICATIONE ECCLESIAE Pap. I, 10, I Bl. 290 × 205. Süddeutscher Raum, Ende 14. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Brixen, Priesterseminarbibl., Cod. A.15 und Cod. A.22. Lage: 1I (Vorsatzbl. 18. Jh.) + V10 + 1I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am Lagenanfang Kustode 22, am oberen Blattrand Foliierung 15. Jh. 232–241, Hinweise auf ehem. Sam melhandschrift (s. bei G). Daneben Foliierung 19. Jh. durch die ULB Tirol 1–10 (hier berücksichtigt). S: Schriftraum 215 × 150. Zwei Spalten, von Tintenlinien gerahmt, zu 40–48 Zeilen. Zirkelstiche. Bastarda. A: Schwarze Unter- und Durchstreichungen sowie Überschriften. Bl. 3rb einfache Federzeichnung eines Ge sichtes im Profil. Platz für zwei- und dreizeilige Lombarden freigelassen, diese jedoch nicht ausgeführt. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Gold bordüren gesäumt. Im zweiten Feld Golddruck M. S., im dritten Signaturschild (s. bei G). Schnitt rot gesprenk elt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Neustift, Stiftsbibl., Cod. 158, Bl. Ir gehört Cod. 162 zu einer Gruppe von urspr. tw. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzel handschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. 178 Cod. 162 Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 10 (Tinte, Bleistift) und N. 162 (Tinte) sowie Bleistiftnotiz Vide Grundzettel. Am Rücken gelbes Signaturschild der ULB Tirol 162. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 138, 206. (Ir) leer. (Iv) Bleistiftnotiz 19. Jh.: Sermones Sacri. (1ra–10rb) Sermones in dedicatione ecclesiae. (1ra) Inc.: [V]enit enim filius hominis querere et salvum facere quod perierat (Lc 19,10). Ista scribit beatus Lucas et sunt verba nostri salvatoris suam dileccionem quam ad quendam humanum habuit … – Expl.: … omnia taliter custodire ut mereamur in templum glorie tue eternaliter introire. (1va) Tit.: Alius sermo de dedicacione templi. – Inc.: [Z]achee festinans descende (Lc 19,5). Nota quod quidam Deum recipiunt in domum suam et non in animam quidam in animam et non in domum quidam in animam et in domum quidam nec in animam nec in domum … – Expl.: … Iudas qui desperavit. R ogemus. Inc. mit geringen Abweichungen wie Schneyer 9, 536, Nr. 77 (zu Stuttgart, LB, theol. fol. 332). (2rb) Tit.: Alter sermo in dedicacione. – Inc.: [B]eati qui habitant in domo tua domine (Ps 84,5). Davit (!) dicit hec verba. Notandum est quod domus domini est quadruplex scilicet domus materialis … – Expl.: … ut mereamur recipi in domum celestem. Rogemus. Inc. wie München, UB, fol. 133, Bl. 162va. (2va) Tit.: In dedicacione. – Inc.: [I]ngressus Ihesus ambulabat Iericho (Lc 19,1). Iericho interpretatur luna et signat mundum propter suum defectum … – Expl.: … nos intrare in regnum celorum. Rogemus. Inc. wie Schneyer 8, 608, Nr. 2 (zu München, BSB, Clm 5998), Schneyer 9, 537, Nr. 86 (zu Stuttgart, LB, theol. fol. 332) oder Schneyer 9, 830, Nr. 13 (zu Vorau, Stiftsbibl., Cod. 161). (3ra) Tit.: De eo sermone in dedicacione. – Inc.: [T]emplum hoc sanctum ingredientes rex pie exaudi nos domine. Nota tria templa placencia Deo. Primum templum est aureum secundum est eburneum tertium est lapideum … – Expl.: … Ite maledicti in ignem eternum. Rogemus. (3va) Tit.: De dedicacione templi. – Inc.: [V]idit Iacob in sompnis scalam et angelos Dei descendetes (!) et ascendentes per eam (Gn 28,12). Ait vere dominus est in loco isto et ego nesciebam (Gn 28,16). Hiis verbis possumus quattuor notare. Primo quis sit Iacob secundo quod dormivit tercio que sit scala quarto quod angelos vidit … – Expl.: … bonam dedicacionem in corde suo habuit. Rogemus. (4ra) Inc.: [E]gressus Ihesus perambulabat Iericho (Lc 19,1). Zacheus interpretatur iustus sed quia ille iustus non erat ideo dominum videre non poterat. Sunt autem tres cause quare dominum videre non poterat. Primo propter divicias … – Expl.: … concessit unde rogemus unum de festo Pentecosten. Refulsit sol quere in fine sermonis. (4va) Inc.: [D]uodecim sunt signa scilicet aries thaurus … Notandum quod in hiis duodecim signis sol perficit cursum suum … – Expl.: … in cuius signis sol perficit cursum simul etc. (5rb) ‹Conradus de Brundelsheim› (Schneyer 1, 746, Nr. 378). (6va) Inc.: [N]ova veniens e celo nupciali thalamo. Apok. XIX (vgl. AH 51, Nr. 102, Str. 2). Nota quod sponsa Christo debet se preparare ad eius adventum quinque modis. Primo se lavando … – Expl.: … ut voluntatem eius faciat. Rogemus dominum ut ab hostibus protegat et ad regnum eternum perducat etc. (7rb) Inc.: [V]enit filius hominis querere et salvum facere. Luc. 19 (10). Hic sumat peccatrix anima confidenciam tripliciter. Quia Christus venit quesivit et salvum fecit venit nascendo quesivit (gestrichen: nasciendo) predicando salvum fecit moriendo … – Expl.: … neque secundum peccata nostra facias nobis in adultera. Rogemus. 179 Cod. 163 Inc. wie Augsburg, Staats- und Stadtbibl., 2o Cod. 420, Bl. 103va. (7va) Inc.: [E]t querebat videre Ihesum quis esset (Lc 19,3). Hec verba scribit Lucas de quodam Iudeo qui vocabatur Zacheus qui desiderabat videre filium Dei … – Expl.: … Gaudium vestrum nemo tollet a vobis ad quod nos perducat. (8ra) Bibelzitat und Expl. wie ‹Aldobrandinus de Cavalcantibus› (Schneyer 1, 212, Nr. 836). Inc.: [T]emplum Dei sanctum est quod estis vos si quis autem templum Dei violavit disperdet illum dominus Deus (1) Chor. 3 (17). In verbis istis apostolus duo facit, ostendit enim fidelis anime sanctitatem et execratur sanctitatis violatorem … (8vb) Inc.: [V]idi civitatem sanctam Iherusalem. Apok. 21 (2). Dicit beatus Augustinus quod duplex est civitas … – Expl.: … cum dicit Vidi civitatem iusticie et iustorum. (9ra) Inc.: Vidi civitatem sanctam etc. (Apc 21,2). Quantum ad statum penitencie ostendit autem civita tem iustorum laudabilem duplici respectu … – Expl.: … hic manentem civitatem festinemus ingredi in futuram civitatem ad quam nos perducat. (9vb) ‹Martinus Oppaviensis› (Schneyer 4, 148, Nr. 319, vgl. auch Schneyer 9, 621, Nr. 57). (10va–b) nur Rahmung. (Ir–v) leer. P. A. / U. S. Cod. 163 ARISTOTELES Pap. I, 79, I Bl. 290 × 220. Wien, T. I: um 1389 und um 1443, T. II: Ende 14. Jh. B: Zwei Teile: I (I, 1–30), II (31–79, I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. DE ANIMA ANTICI (!) in Goldrahmung, im drit ten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Innsbruck, ULB Tirol, Cod. 157, Bl. Ir gehört Cod. 163 zu einer Gruppe von urspr. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wur den. Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signa turen (in der urspr. Reihenfolge): Cod. 157: Aristoteles: Ethica Nicomachea und Oeconomica. heute nicht mehr nachweisbar: Aristoteles: Politica. Cod. 159: Albertus de Saxonia: Kommentar zu Cod. 157. Cod. 168: Aristoteles: Physica, Lib. 1–5. Cod. 163: Aristoteles: Physica, Lib. 6–8 und De anima. Cod. 181: Alanus ab Insulis: Summa de arte praedicatoria. Cod. 164: Anshelmus Havelburgensis. Anticimenon. Von diesen sieben Hs. ist eine nicht mehr erhalten, drei befinden sich heute an der ULB Tirol in Innsbruck (Cod. 157, 159 und 164), drei werden in der Stiftsbibl. Neustift (Cod. 163, 168 und 181) aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 2 H 11 (Tinte, Bleistift) und Nr. 163 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 163. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. 180 L: Cod. 163 Lacombe II 1023, Nr. 1495. – Neuhauser, Neustift 91–95. – Katalog ULB Tirol II 138, 206. – W. Neuhauser, Buchgeschichtliche Beziehungen zwischen dem Wiener Raum und Tirol im Mittelalter. Jahrbuch für Landeskunde von Niederösterreich N. F. 63/64 (1997/98) 57 (auch in: Neuhauser, In libris 405–424). I (I, 1–30) B: Bl. 2–5 und 8–10 lose. Bilder und Nachweise der Wasserzeichen s. WZMA. Bl. 1–24 Datierung nach Was serzeichenbefund. Identische Wasserzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 477, T. II (Korneuburg, dat. 1389) und Wien, Schottenstift, Archiv, Hs. 273, T. I (Prag, dat. 1389). Bl. 25–30 wohl später eingefügt: Datierung um 1443 nach Wasserzeichenbefund. Identische Wasserzeichen: Klosterneuburg, Stiftsbibl., CCl 369 (dat. 1443). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 2.VI24 + III30. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Zw. Bl. 15 und 16 ungezähltes kleines Bl. (135/145 × 110/115) eingefügt. Bl. 1–29 am oberen Blattrand der Rectos zeitgenössische Buchzählung. Lagenreklamanten. S: Text von Kommentar umgeben. Schriftraum Haupttext 160 × 100/105, von Tintenlinien gerahmt, zu 27–34 Zeilen. Mehrere Hände: 1) Bl. 1r–24v: Bastarda 14. Jh. von zwei sich abwechselnden Händen; 2) Bl. 25r– 29r Bastarda 15. Jh. Schriftraum der Kommentarfelder stark wechselnd, von Tintenlinien gerahmt, biswei len bis zum Blattrand beschrieben: oberes Kommentarfeld 40 mm, unteres 65 mm hoch, äußeres 65/70 mm (tw. beschnitten, Textverlust), inneres 30 mm breit. Vgl. auch Cod. 168. A: Rote Auszeichnungsstriche, Unterstreichungen, Rahmungen sowie schwarze, rot konturierte Paragraph zeichen und Zeigehände. Zwei- bis sechszeilige rote Lombarden mit Punktverdickungen; vereinzelt zwei zeilige schwarze, rot umrahmte Lombarden mit etwas Dekor. Repräsentanten. 1 (Ir–v) leer. (1r–29r) Forts. von Cod. 168, Bl. 46v: Aristoteles: Physica, Lib. 6–8, Translatio nova durch Guilelmus de Moerbeke (Druck: u. a. GW 2336. Ed.: Aristoteles Latinus VII: Physica, Fasc. 1: Translatio vetus, ed. F. Bossier, J. Brams, rev. ed. A. Mansion. Leiden, New York 1990. – La combe I 52, Nr. 17 und I 126–127, Nr. 17 sowie II 1023, Nr. 1495. Thorndike–Kibre 1296). Mit umfangreichem Kommentar. (1r–9v) Lib. 6. – (10r–14v) Lib. 7. – (15r–29r) Lib. 8. (24v) Expl. mut.: … infinito tempore infinitum transiit aliquis similiter autem [ Bl. 25r–v leer ohne Textverlust. Text setzt Bl. 26r von anderer Hand unmittelbar fort. Inc. mut.: ] infinitum utique (!) in longitudine est et in tempore … (29r) Expl. (leicht abweichend): … manifestum utique quod indivisibile est et impartibile et nul lam habens magnitudinem Amen. (29v–30v) leer. (1r–29r) Kommentar. (1r–9v) Zu Lib. 6. Inc.: Iste est liber sextus Phisicorum, in quo philosophus determinat de divi sione motus et quietis in suas partes … – Expl.: … sed non eedem semper et cetera autem finis sexti libri Phisicorum Deo gracias. (10r–14v) Zu Lib. 7. Inc.: Omne quod movetur necesse. Iste est septimus liber Phisicorum in quo philosophus determinat de comparatione motuum … – Expl.: … est finis huius septimi libri de quo sit Iesus Marie filius in secula seculorum benedictus Amen. (15r–29r) Zu Lib. 8. Inc.: Utrum autem factus. Iste est octavus liber Phisicorum in quo Aristoteles determinat de perpetuitate motus … – Expl.: … de rebus naturalibus in principio totius nature qui est super omnia benedictus Deus in secula Amen. Lit.: Grabmann, Aristoteles 170–174. Cod. 163 181 II (31–79, I) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Innsbruck, ULB Tirol, Cod. 221 (dat. 1390); Wien, Schottenstift, Archiv, Hs. 367 (dat. 1391) und Archiv, Hs. 362 (dat. 1400); Korneuburg, Stadtarchiv, Cod. 1756 (dat. 1396). Lagen: (VI+1)43 + 3.VI79 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Bl. 31 zusätzlich eingefügtes halbes Bl. (290 × 110/115). Lagenreklamanten, tw. weggeschnitten. S: Text von Kommentar umgeben (Bl. 75r–76v ohne Kommentar). Schriftraum wechselnd: Haupttext Bl. 31r– 75r und 78r–79v: 150 × 85/95, Bl. 75v–77v: 185/210 × 125/140, von zweifachen Linien gerahmt (Bl. 75v– 76v nur seitlich einfach begrenzt), zu 16–20 Zeilen, Bl. 75r zu 21 Zeilen, Bl. 75v zu 23 Zeilen, Bl. 76r–v zu 27 Zeilen. Vier Hände Ende 14. Jh.: 1) Bl. 32r–44r, Z. 3 Textualis cursiva; 2) Bl. 44r, Z. 4–74v Bastarda; 3) Bl. 75r–76v kursive Bastarda von späterer Hand; 4) Bl. 79v flüchtige Notula von späterer Hand in dunkler Tinte, geschrieben wohl in Neustift, vgl. die Eintragungen in Innsbruck, ULB Tirol, Cod. 157, 159 und 164. Randglossierung von mehreren Händen Ende 14. Jh.: 1) Bl. 31r–34v Notula; 2) Bl. 35r–36r Bastarda (spä tere Hand), jeweils beginnend Notandum …; 3) 47r–48v Bastarda (gleicher Schreiber wie Bl. 35r–36r); 4) Bl. 50v–74v Bastarda. Zusätzlich Bl. 32r–37v, 47r–49r Interlinearglossierung, danach nur vereinzelt. A: Rote Auszeichnungsstriche, Unterstreichungen, Rahmungen sowie schwarze, rot konturierte Paragraph zeichen und Zeigehände. Ein- bis vierzeilige rote Lombarden, Bl. 47r und 48v mit gegenständigen Halb palmetten im Binnenfeld. Bl. 75r–76v ohne Rubrizierung. 2 (31r–v) Forts. des Kommentars von Bl. 32r. (32r–76v) Aristoteles: De anima, Translatio nova durch Guilelmus de Moerbeke (Druck: GW 2343–2349. Aristotelis de anima libri tres, cum Averrois commentariis et antiqua tralatione suae integritati restituta … Michaele Sophiano interprete. Venetiis 1562, ND Frankfurt a. M. 1962. – Lacombe I 58–59, Nr. 27 und I 136–137, Nr. 27, sowie II 1023, Nr. 1495. Thorndike–Kibre 179). Mit Kommentar bis Bl. 75v und tw. Interlinearglossen. Abweichender Beginn von Lib. 3 (71v) De parte autem anime quam cognoscit anima et sapit … (vgl. Lacombe I 58–59) Die im Druck 1562 als „antiqua tralatio” bezeichnete Übersetzung entspricht der Translatio nova bei Lacombe. Text. Inc.: Bonorum honorabilium notitiam opinantes magis autem alteram altera aut secundum certitudinem aut ex eo … – Expl. mut. (Lib. 3, cap. 10): … videtur enim quod iam delectabile et simpliciter delectabile et bonum simpliciter [ (32r) Lib. 1. – (47r) Lib. 2. – (71v) Lib. 3. (31r–34v) Kommentar 1. Inc. (Bl. 32r): Circa inicium huius libri sicud aliorum sunt aliqua generalia autem textus in tytulationem prenotanda. Primo in quali sciencia … Forts. nach Bl. 32r auf Bl. 31v und anschließend auf Bl. 31r, Forts. auf Bl. 32v. – Expl.: … ut benedicam ab eis acceptemus et alium refutemus. (35r–36r) Kommentar 2. Inc.: Notandum circa textum nonnulli priorum oppinati (!) sunt quod quando anima ingrediebatur corpus… – Expl.: … Quidam dicunt Platonem asseruisse animam esse iterum se ipsam moventem male dicunt hec opinio nomine Platonis secundum Temocritum. (47r–48v) Kommentar 3 (zu Lib. 2). Inc.: Circa textum notandum quod philosophus in textu dicit quod erit determinandum … – Expl.: … Tonitruum est sonus factus in nubibus propter extinc tionem inflammacionis in eisdem. (50v–71r) Kommentar 4a (zu Lib. 2). Inc.: Doctrinarum autem anime. Capitulum 3m in quo de terminatur de potenciis anime … – Expl.: … racione sanatur immediate ministrat in temporali. Intenciones sensibilia ad hoc igitur deservit (!) sanacionem. (71v–74v) Kommentar 4b (zu Lib. 3). Inc.: De parte autem anime. Iste est tercius liber parcialis in quo dicitur de aliis anime potenciis … – Expl.: … est posterius considerandum scilicet in 2o idem in 12o Methaphysice. Lit.: Grabmann, Aristoteles 190–198. (77r–79r) nur Rahmung. 182 3 Cod. 165 (79v) Notabilia zu De anima. Inc.: Item nota quod per quiditatem anime intellegitur summa et essencia ipsius anime … – Expl.: … Alio modo potest considerari prout inseparata a corpore 2o de ipsa determinare potest ad mathematicam. (Ir–v) leer. P. K. / W. N. Cod. 165 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. I, 148, I Bl. 295 × 205. Süddeutscher Raum, 1. Jz. 15. Jh. (T. III: 1407). B: Drei Teile: I (I, 1–74[75]), II (75[76]–113[115]), III (114[116]–146[148], I). Bilder und Nachweise der Wasserzeichen s. WZMA. T. III dat. 1407, Wasserzeichen von T. III auch in T. I und II. Identische Wasser zeichen: u. a. Neustift, Stiftsbibl., Cod. 166. In der Mitte des oberen Blattrandes (am unteren Blattrand meist wiederholt) Foliierung 59–205 (bisweilen beschnitten), urspr. Bl. 1–58 fehlen. Am rechten oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol 1–145; Fehler in der Zählung: 21, 86 und 97 doppelt angeführt, 87 ausgelassen. Neuzeitliche gelbe Papiersignakel der ULB Tirol. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit Leder überzogen. Rücken: braunes Leder, drei einfache Bünde, von Goldbor düren gesäumt. Im zweiten Feld Golddruck OPUSCULA DIVERSOR. PATRUM in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Die zeitgenössische Foliierung in der Mitte des oberen Blattrandes lässt – gemeinsam mit den übereinstimmenden Wasserzeichen sowie der Größe der Hs. – darauf schließen, dass Cod. 165 urspr. mit Cod. 166 vereinigt war. Gegebenenfalls wohl im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halb ledereinband. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 2 H 13 (Tinte, Bleistift) und N. 165/(8) (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 165/(8). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat ab gegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 4819,4 Suppl. (zu Bl. 129–132). – Katalog ULB Tirol II 140, 206–207. – Katalog ULB Tirol II, Beiheft, Taf. 52. – F. Eisermann, „Stimulus amoris“. Inhalt, lateinische Überlieferung, deutsche Übersetzungen, Rezeption (MTU 118). Tübingen 2001, 153, 214, 224, 324, 327. I (I, 1–74[75]) B: Pap. fleckig. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 4.VI47(48) + (V+1)58(59) + VI70(71) + II74(75). Bl. 52 eingehängtes Einzelbl. Am oberen Blattrand der Rectos Zählung der einzelnen Bücher (Abschnitt 1) bzw. Überschriften (Abschnitt 3–5), bisweilen beschnitten. S: Schriftraum 230 × 150. Zwei Spalten, von tw. Bleistift-, tw. Blindlinien gerahmt, zu 50 (Schreiber 1) bzw. 41–42 (Schreiber 2) Zeilen. Bastarda, Anfang 15. Jh. von zwei Händen: 1) Bl. 1ra–58vb (= Cod. 166, Schreiber 1); 2) Bl. 59ra–74rb. A: Rubrizierung von unterschiedlichen Händen: 1) Bl. 1ra–58vb; 2) Bl. 59ra–74rb. Rote Auszeichnungsstriche, Paragraphzeichen, Zeilenfüllsel, Unterstreichungen, Über- und Schlussschriften. Zeigehände. Rote Satz majuskeln und ein- bis dreizeilige Lombarden; Bl. 42va dreizeilige Initiale P in roter Umrisszeichnung, schwarz konturiert und mit schwarzem Fleuronnée (Knospengarben im Binnenfeld, gepunktete Perlen als Besatz). Cod. 165 183 1 (1ra–42ra) Gregorius Magnus: Dialogi = Dialogorum libri IV de vita et miraculis patrum Itali corum (PL 77, 149–429 [Lib. 1, 3 und 4] bzw. PL 66, 126–204 [Lib. 2]. Ed. U. Moricca [Fonti per la storia d’Italia 57]. Roma 1924. Ed. A. de Vogüé [Sources chrétiennes 260]. Paris 1979 [Buch 1–3] und [Sources chrétiennes 265]. Paris 1980 [Buch 4]. – CPL Nr. 1713). (1ra) Lib. 1. – (8va) Kapitelübersicht zu Lib. 2. – (8vb) Lib. 2 = Vita s. Benedicti abbatis. – (16vb) Kapitelübersicht zu Lib. 3. – (17ra) Lib. 3. – (28ra) Kapitelübersicht zu Lib. 4. – (28va) Lib. 4. (42ra) Schlussschrift: Explicit Dyalogus Gregorii. 2 (42ra) Exzerpte aus ‹Ps.-›Augustinus: Auctoritates. Tit.: Diffinicio virtutis secundum beatum Augustinum. – Inc.: Est amor ordinatus videlicet ut amet … – Expl.: … alii gustum reparat, alii aliud et aliud operatur etc. Parallelüberlieferung: u. a. Fiecht, Stiftsbibl., Hs. 15, Bl. 82r. Darunter von späterer Hand: Qui non est dives et nescit vivere pauper / Ille modum vite nescit habere sue (Walther, Proverbia Nr. 24397. Vgl. Walther, Initia Nr. 15581). (42rb) leer. 3 (42va–53va) ‹Ps.-›Eusebius Caesariensis: Epistola ad beatum Damasum episcopum Portuensem et ad christianissimum Theodosium Romanorum senatorem de morte Hieronymi (PL 22, 239– 282. Klapper II 10–241. – BHL Nr. 3866. BHM Nr. 903E). Tit.: Incipit epistola beati Ewsebii ad Damasium episcopum (Nachtrag: Portuensem) et Theodo nium (!) Romanorum senatorem sui magistri gloriosissimi sancti Ieronimi (gestrichen: episcopi Portuensis) presbiteri. Schlussschrift: Explicit epistola Ewsebii de morte sanctissima Ieronimi presbiteri. 4 (53va–55vb) ‹Ps.-›Augustinus: Epistola 18 (ad Cyrillum Hierosolymitanum episcopum de agnificentiis beati Hieronymi) (PL 22, 281–289. Klapper II 245–288. – CPL Nr. 367. BHL m Nr. 3867. BHM Nr. 903A). Tit.: Incipit epistola beati Augustini ad Cyrillum patriarcham Ierosolitanum de beatissimo Iero nimo. Exordium epistolare de non tacendo laudem Ieronimi. Schlussschrift: Explicit epistola Augustini ad Cyrilium de glorioso Ieronimo. 5 (55vb–71rb) ‹Ps.-›Cyrillus: Epistola ad Augustinum de miraculis Hieronymi (PL 22, 289–326. – CPL Nr. 367. BHL Nr. 3868. BHM Nr. 903C) = ‹Ps.-Augustinus›: Epistola App. 19 (PL 33, 1126–1153). Tit.: Incipit epistola Cyrilli ad Augustinum de miraculis gloriosi Ieronimi. 6 (71rb–74rb) Auszüge aus ‹Iohannes Andreae›: Hieronymianus (Druck: GW 1727. – BHL Nr. 3876. BHM Nr. 907). (71rb–73rb) Tit.: De quibusdam miraculis vel mirabilibus factis noviter in Apulia et primo quid in Troia civitate etc. – Inc.: Sciendum est quod moderno tempore in Apulia in provincia Capiti mate (!) … – Expl.: … fidei et gremium ecclesie sunt reversi. Deo gracias. (73va–74rb) Inc.: Hic s. Ieronimus sacerdos fuit ordine peregrinacionis … Frater Iacobus de varietatibus legendarum … – Expl.: … rex Israheliticus occidente sole. (74rb) Schlussschrift: Explicit libellus de vita et obitu et miraculis sancti Ieronimi doctoris egre gii. Ergo sancte Ieronime pro me misero peccatore intercede ad Ihesum Christum qui propter nos venit (gestrichen: ad) in mundum istum etc. (74va–b) nur Rahmung. Zu Texten und Anordnung von T. I vgl. u. a. Wilten, Stiftsbibl., Cod. 32 02 05, T. IV. Zu Nr. 3–6 vgl. Cod. 125, Bl. 170rb–200ra (bes. Nr. 10). Lit.: Schulte GQ II 217. 184 Cod. 165 II (75[76]–113[115]) B: Pap. fleckig. Lagen: 2.VII101(103) + VI113(115). S: Schriftraum 230 × 150. Zwei Spalten, von meist Blind-, tw. Bleistiftlinien gerahmt, zu 50 Zeilen. Bastarda Anfang 15. Jh. von zwei Händen: 1) Bl. 75ra–111ra, Z. 46 (= Schreiber 1 von T. I); 2) Bl. 111ra, Z. 46– Bl. 113va. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unter- bzw. Durchstreichungen, Überschriften. Zeigehände. Rote Satzmajuskeln sowie ein- bis dreizeilige Lombarden. 7 (75ra–88va) ‹Rufinus Aquileiensis›: Historia monachorum = Vitae patrum, Lib. 2 (Druck: PL 21, 387–462. Ed.: E. Schulz-Flügel, Tyrannius Rufinus, Historia monachorum sive de vita sanctorum patrum [Patristische Texte und Studien 34]. Berlin u. a. 1990, 234–387.– CPL Nr. 198p. BHL Nr. 6524). (75ra) Tit.: In Christi nomine incipit liber de vita sanctorum patrum Heremitarum. (75ra) Register. (75ra) Prolog. (75va) Cap. 1: De s. Iohanne. – (78vb) Cap. 15: De Apellen presbytero et Iohanne. – (79rb) Cap. 16: De Paphnutio. – (80rb) Cap. 12: De Elia. – (80rb) Cap. 13: De Pithyrione. – (80va) Cap. 14: De patre Eulogio. – (80va) Cap. 7: De Apollonio. – (82va) Cap. 8: De Ammone. – (83ra) Cap. 9: De Coprete presbytero et patermutio. – (84va) Cap. 11: De Heleno. – (85rb) Cap. 6: De Theone. – (85va) Cap. 20: De Dioscuro presbytero. – (85va) Cap. 2: De Hor. – (86ra) Cap. 5: De Oxyryncho civitate. – (86rb) Cap. 18: De Serapione presbytero. – (86va) Cap. 19: De Apollonio monacho et martyre. – (86vb) Cap. 21: De monachis in Nitria commorantibus. – (86Ira, Z. 20) Cap. 22: De loco qui dicitur Cellia. – (86Ira) De Ammonio (bis PL 21, 446B: … puteum foderat, Bl. 86Irb, Z. 23). – (86Irb, Z. 23) Cap. 24: De Didymo. – (86Irb) Cap. 30: De Ammone, primo Nitriae monacho (bis PL 21, 456C: … sanitas redditur). – (87va) Cap. 31: De Paulo simplice. – (88ra) Cap. 32: De Piammone presbytero. – (88rb) Cap. 33: De Iohanne. – (88rb) Epilog. (88va) Schlussschrift: Explicit liber primus. 8 (88va–104vb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3 = Verba seniorum (Druck: GW M50865. PL 73, 739A–808B. – BHL Nr. 6525). (88va) Register. (90ra) Prolog. Tit.: Incipit liber secundus de vita sanctorum patrum prologus. (90ra) Text. Tit.: De odio mundi. (104vb) Schlussschrift: Explicit liber secundus. 9 (104vb–111ra) ‹Sulpicius Severus, Iohannes Cassianus›: Vitae patrum, Lib. 4 = Excerpta (PL 73, 813–842A. – BHL Nr. 6526). (104vb) Register. (105rb) nur Rahmung. (105va) Tit.: Liber tercius. – Expl.: … quam dominus ab eo avertere voluit ipse sibi conscivit (= cap. 44). (111ra, unten) Vermerk: cetera capitula require in dialogum in isto signo x in chilem non credas. 10 (111rb–113va) Notiz. Tit.: Nota. – Inc.: In hystoria ecclesiastica habetur de institucione vite sanctorum patrum in Egypto … – Expl. mut.: … quasi absque tumultu populi necari [ (113vb) nur Rahmung. Cod. 165 185 III (114[116]–146[148], I) B: Bl. 114 abgegriffen (wohl urspr. ohne Einband). Lagen: 2.VI137[139] + (III+3)146[148] + (I–1)I* (Nachsatzbl. 18. Jh.). Bl. 138–140 eingehängte Einzelbl. S: Schriftraum 240 × 160, von Tintenlinien gerahmt, zu 50 Zeilen. Bl. 143v–146v: Drei Spalten, von Bleistift linien gerahmt. Bastarda 15. Jh. von zwei Händen: 1) Bl. 114r–143r; 2) Bl. 143v–145v, dat. Juni 1407 (Bl. 128v). A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Rahmungen, Über- und Schlussschrif ten. Zeigehände (z. B. Bl. 126v mit Ärmelansatz). Rote zweizeilige Lombarden. 11 (114r–128v) Quaestiones disputatae de peccato in spiritum sanctum. Tit.: De peccato in spiritum sanctum et misericordia Dei. Darüber Schreibervers: Assit ad incep tum sancta Maria meum. Inc.: Dubitatur numquid aliquid peccatum sic (!) tam grande … – Expl.: … tamquam verus ami cus. Sic est finis. (128v) Schlussschrift und Datierung: Expliciunt questiones bone disputate a diversis doctoribus in studio Pragensi anno domini millesimo quadicentesimo (!) septimo mensis Iunii etc. (Juni 1407). Daneben von späterer Hand in anderer Tinte: 1407. Parallelüberlieferung: u. a. München, BSB, Clm 28564, Bl. 246ra. 12 (129r–131v) Determinatio doctorum universitatis Pragensis de V propositionibus magistri Iohannis Muntzinger 1386 facta = Gutachten der Prager theologischen Fakultät im Rahmen der theologischen Streitigkeiten zw. Johannes Müntzinger und den Dominikanern in Ulm (Druck: J. G. Schelhorn, Amoenitates literariae … VIII. Frankfurt, Leipzig 1728, 527–552. – Stegmüller RB Nr. 4819,4 Suppl., mit Nennung der Hs.). Inc.: In exordio presentis negocii Deo scienciarum domino humiliter invocato protestamur quod circa puncta nuper in forma publice scripture de oppido Ulmensi Constantiensis diocesis trans missa … – Expl.: … probabile reputamus. Lit.: A. Lang, Johann Müntzinger, ein schwäbischer Theologe und Schulmeister, in: Geisteswelt des Mittelalters. Studien und Texte. Martin Grabmann zur Vollendung des 60. Lebensjahres von Freunden und Schülern gewidmet. München 1935, 1208–1210; A. Holtorf, „Müntzinger (Mün zinger), Johannes“. Verfasserlexikon2 VI 794–799. 13 (131v–132r) Quaestio de articulis Iohannis Müntzinger. Inc.: Utrum solus Deus sit adoracione latrie adorandus … – Expl.: … sanguinem circumcisionis non creditur resumpsisse. Sic est finis. In Fritzlar, Dombibl., Ms. 6, Bl. 296vb–297rb Conradus de Soltau zugeschrieben. Schlussschrift (zu Nr. 12 und 13): Expliciunt punctus magistri Iohannis Munczinger. 14 (132v–139v) ‹Iacobus Mediolanensis› (=‹Ps.-Bonaventura›): Stimulus amoris, Auszug (Druck: A. Ch. Peltier, S. R. E. Cardinalis S. Bonaventurae … opera omnia XII. Parisiis 1868, 633–703. Stimulus amoris fr. Iacobi Mediolanensis. Canticum pauperis Fr. Ioannis Peckam [Bibliotheca Franciscana Medii Aevi 4]. Quaracchi 21949, 1–129. – Distelbrink Nr. 217–219. Bloomfield Nr. 1294). Die Hs. enthält Pars 1, cap. 1–4; Pars 2, cap. 1, 3–6, 9, 10; Pars 3, cap. 1–5 des Drucks (1868). Rubrik: Liber iste qui intitulatur stimulus amoris in dulcissimum et piissimum Ihesum salvatorem nostrum in tres dividitur partes … Inc.: Currite gentes undique et miramini erga vos caritatem Dei … – Expl.: … misericors est Deus qui est benedictus et laudabilis et gloriosus in secula seculorum Amen. Schlussschrift: Explicit stimulus amoris. 186 Cod. 166 Vgl. Cod. 125, Bl. 161ra. Parallelüberlieferung (mit derselben Auswahl): Innsbruck, ULB Tirol, Cod. 140, Bl. 145va. Lit.: F. Eisermann, „Stimulus amoris“. Inhalt, lateinische Überlieferung, deutsche Übersetzungen, Rezeption (MTU 118). Tübingen 2001 (mit Nennung der Hs.). 15 (139v–141r) ‹Ecbertus Schonaugiensis›: Soliloquium seu Meditationes (PL 195, 105D–114). Tit.: Contemplacio Bernhardi. Schlussschrift: Explicit contemplacio beati Bernhardi. Vgl. Cod. 125, Bl. 169va. 16 (141r–143r) Augustinus: Sermo 9 de decem chordis (PL 38, 75–91. CCSL 41, 105–151. CPPM I 456). Tit.: Augustinus de decem cordis. 17 (143va–144ra) De forma vivendi monachorum (AH 33, Nr. 220). Inc.: Christus nobis tribuit normam (!) hic vivendi / Et exemplo docuit viam gradiendi / Ut mun dana vanitas possit vilipendi … – Expl.: … qui timent turbamen / Et a malo liberat (gestrichen: liberit) in eternum Amen. Darunter zwei Strophen hinzugefügt (vgl. AH 33, Nr. 220, S. 207): Qui transitis monachi omnes huc venite … Sed perlege iugiter atque meditare. 18 (144rb–vc) Claustrum animae. Tit.: Nota quoddam claustrum spirituale. – Inc.: Manus Zorobabel fundaverunt eam (Za 4,9) scilicet animam per istum Zorobabel eam … – Expl.: … ad triclinium trinitatis. Quod nobis dignetur concedere anime verus sponsus Iesus Christus dominus noster qui cum patre et spiritus sanctus vivit et regnat per omnia secula seculorum Amen. Vgl. u. a. Innsbruck, ULB Tirol, Cod. 73, Bl. 113ra. (145ra–146vc) nur Rahmung. (Ir–v) leer. C. S. Cod. 166 SPECULUM HUMANAE SALVATIONIS Pap. II, 58, I Bl. 295 × 210. Süddeutscher Raum, 1. Jz. 15. Jh. B: Pap. bes. am Beginn stark verschmutzt, Bl. 1 und 2 beinahe lose, Bl. 11 lose. Blätter bisweilen mit schmalen Papierfälzen zur Verstärkung umschlagen bzw. geklebt. Bl. 1 urspr. mit Papierfalz auf Bl. I aufgeklebt, heute gelöst. Bl. II (Einzelbl., 130 × 190) an Bl. 1r angeklebt. Nachweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Neustift, Stiftsbibl., Cod. 165 (T. III dat. 1407); Wien, Schottenstift, Archiv, Hs. 102 (dat. 1408). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 1II + 2.VI24 + V34 + 2.VI58 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am Lagenende Kustoden in arabischen Ziffern, meist beschnitten. Am unteren, tw. zusätzlich am oberen Blattrand zeitgenössische Foliierung 1–10, 35 sowie 50–58, am oberen Blattrand fehlende Foliierung im 19. Jh. durch die ULB Tirol ergänzt. Bl. 52 neuzeitliches oranges Papiersignakel der ULB Tirol. S: Schriftraum 215/220 (Bl. 3–71r: 125/135) × 170/175. Oberes Drittel des Schriftraums Federzeichnungen vorbehalten. Zwei Spalten, von feinen Tinten-, ab Bl. 35 meist von Blindlinien gerahmt, zu 41–51 Zeilen, wenn Federzeichnungen zu 25–28 Zeilen, bisweilen auf Tinten- bzw. Blindlinierung. Zirkelstiche. Bastarda von zwei Händen: 1) Bl. IIv–52rb (= Cod. 165, Schreiber 1); 2) Bl. 52va–56ra. Beischriften zu den Feder zeichnungen wohl von den Schreibern mit unterschiedlicher Feder. Cod. 166 A: 187 Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Über- und Schlussschriften; Bildbei schriften tw. (schwarz-)rot gerahmt; Bl. 3rb als Kolumnenabschluss Punktblüten auf roter Wellenlinie; Bl. 14ra der thronende Pharao baldachinartig in Rot (von Beischriftenhand) umkritzelt. Ein- und zweizeilige rote Lombarden und I-Versalien, Bl. 52vb–55vb fallweise mit freien Punkten. Repräsentanten. Zu den 45 Kapiteln des Speculum humanae salvationis 192 Federzeichnungen, tw. koloriert (dazu s. u.). Typologische Ausdeutung der Vita Christi, deren Bildprogramm sich – eingeleitet von der Schöpfungs geschichte (cap. 1–2) und abgeschlossen von den sieben Stationen des Leidens Christi sowie den sieben Schmerzen und Freuden Mariens (cap. 43–45, jeweils acht Bilder) – in 40 Vierergruppen untergliedert; pro Gruppe jeweils ein Antitypus und drei meist dem Alten Testament entnommene Typen; Antitypus jeweils über linker Spalte des Verso, sodass jede Viergruppe 2 : 2 auf einer Doppelseite zu stehen kommt und als Einheit wahrgenommen werden kann. Anfänge der typenbezogenen Textabschnitte meist durch Paragraph zeichen und Marginalvermerke (prima/secunda/tertia figura) gekennzeichnet. Cod. 166 gehört zur sog. Deutschen Gruppe des Speculum humanae salvationis, deren Bildprogramm sich in der 1. H. 14. Jh., zeitnah zur Entstehung dieses in zahlreichen Hs., Blockbüchern und Inkunabeln erhal tenen Erbauungsbuches, entwickelte. Ausführlich zur Ikonographie MeSch II 9–17, Kat. 3 (Wien, ÖNB, Cod. 1636; M. Roland), insbes. 12–15. Bemerkungen zu Abweichungen von der gängigen Ikonographie der Deutschen Gruppe mit * gekennzeichnet. Bildbeischriften (in Schwarz und Rot, von verschiedenen Händen) auf Typen konzentriert; nachfolgend nur zitiert, wenn sie über Namen, Dingbezeichnungen und Angaben von Bibelstellen hinausgehen; [HS]: in Beischrift Historia Scholastica explizit als Quelle genannt. Cap. 1: Bl. 3va Engelsturz. Bl. 3vb Erschaffung Evas. Bl. 4ra Gott vermählt Adam und Eva. Bl. 4rb Versu chung Evas durch die Schlange. Cap. 2: Bl. 4va Sündenfall. Bl. 4vb Vertreibung aus dem Paradies. Bl. 5ra Adam und Eva arbeiten (In sudore wultus tui vesceris pane tuo). Bl. 5rb Arche Noah. Cap. 3: Bl. 5va Verkündigung der Geburt Mariens an Anna (auf dem Spruchband des Engels: Ave gracia). Bl. 5vb Traum des Königs Astyages von der Geburt des Cyrus [HS]. Bl. 6ra Verschlossener Garten und versiegelter Brunnen als schematisierte Darstellung in rotem Kreis. Bl. 6rb Bileam kämpft gegen Engel. Cap. 4: Bl. 6va Mariengeburt: Maria stehend zw. Anna und Joachim (Annas Bett fehlt). Bl. 6vb Wurzel Jesse. Bl. 7ra Verschlossenes Tor des Tempels (Sinnbild Mariens). Bl. 7rb Tempel Salomos. Cap. 5: Bl. 7va Darbringung Mariens im Tempel: Maria als Dreiviertelfigur auf Säule. Bl. 7vb Der goldene Tisch der Sonne in Zabulon (Mensa solis inventa a piscatoribus). Bl. 8ra Jiftach opfert seine Tochter. Bl. 8rb Semiramis in den hängenden Gärten. Cap. 6: Bl. 8va Verlobung Mariens. Bl. 8vb Verlobung von Sara und Tobias. Bl. 9ra Turm des Baris. Bl. 9rb Turm Davids mit Schilden. Cap. 7: Bl. 9va Verkündigung an Maria. Bl. 9vb Brennender Dornbusch (auf tafelartigem Spruchband Got tes: Audi Israel mandata). Bl. 10ra Gideon und das Vlies. Bl. 10rb Rebekka und Eliëser am Brunnen. Cap. 8: Bl. 10va Geburt Christi. Bl. 10vb Traum des Mundschenks des Pharao. Bl. 11ra Aarons grünender Stab. Bl. 11rb Sibylle verkündet Kaiser Augustus die Geburt Christi (u. a. Quere in passionali de nativitate domini). Cap. 9: Bl. 11va Anbetung der Heiligen Drei Könige. Bl. 11vb Die Heiligen Drei Könige erblicken den Stern (u. a. Quere in passionali de nativitate domini). Bl. 12ra Die drei Helden bringen David Wasser. Bl. 12rb Thron Salomos: Salomo als Büste auf Säulenstumpf, vor dem Thron kniet Königin von Saba (ohne Gabe). Cap. 10: Bl. 12va Darbringung Christi im Tempel. Bl. 12vb Bundeslade. Bl. 13ra Siebenarmiger Leuchter (wirkt durch Blattdekor und grüne Kolorierung wie eine Pflanze). Bl. 13rb Darbringung Samuels im Tempel. Cap. 11: Bl. 13va Flucht nach Ägypten, im Hintergrund Götzenbilder (Genesis 27 de Rebeca que Iacob consuluit ut fugent). Bl. 13vb Ägyptische Madonna [HS]. Bl. 14ra Der Knabe Mose greift in die glühenden Kohlen [HS]. Bl. 14rb Traum Nebukadnezars. Cap. 12: Bl. 14va Taufe Christi. Bl. 14vb Ehernes Meer im Tempel (von sechs Rindern getragenes Erz becken). Bl. 15ra Naamans wird durch Bad im Jordan geheilt. Bl. 15rb Israeliten tragen Bundeslade durch den Jordan. Cap. 13: Bl. 15va Dreimalige Versuchung Christi durch den Teufel. Bl. 15vb Daniel tötet den Drachen des Götzen Bel. Bl. 16ra David tötet Goliat. Bl. 16rb David tötet einen Löwen und einen Bären (im Hintergrund zwei kleinere Löwen). Cap. 14: Bl. 16va Maria Magdalena trocknet die Füße Christi mit ihren Haaren. Bl. 16vb Der gefangen ge setzte und büßende König Manasses. Bl. 17ra Heimkehr des verlorenen Sohnes. Bl. 17rb Natan vor David. Cap. 15: Bl. 17va Einzug Christi in Jerusalem. Bl. 17vb Jeremia klagt auf den Mauern von Jerusalem. Bl. 18ra David mit dem Haupt Goliats wird von Michal und Saul ehrenvoll empfangen. Bl. 18rb Geißelung Eliodors. Cap. 16: Bl. 18va Letztes Abendmahl. Bl. 18vb Mannalese. Bl. 19ra Juden braten das Paschalamm. Bl. 19rb Melchisedek und Abraham. 188 Cod. 166 Cap. 17: Bl. 19va Christus wirft die Soldaten, die ihn verhaften wollen, mit seinem Wort nieder. Bl. 19vb Simson erschlägt die Philister mit einem Eselskinnbacken. Bl. 20ra Schamgar tötet 600 Männer mit einer Pflugschar. Bl. 20rb David tötet 800 Männer mit einem Hieb. Cap. 18: Bl. 20va Gefangennahme Christi (Judaskuss, Christus heilt das von Petrus abgeschlagene Ohr des Malchus). Bl. 20vb Joab tötet Amasa. Bl. 21ra Saul attackiert David mit einem Speer. Bl. 21rb Opfer Abels und Kains; Kain erschlägt Abel. Cap. 19: Bl. 21va Verspottung Christi. Bl. 21vb Hur wird zu Tode gespien [HS]. Bl. 22ra Noahs Schande. Bl. 22rb Verspottung (und Blendung) Simsons. Cap. 20: Bl. 22va Geißelung Christi. Bl. 22vb Achior wird an einen Baum gebunden. Bl. 23ra Lamech wird von seinen beiden Frauen verspottet. Bl. 23rb Ijob wird von seiner Frau und vom Teufel gegeißelt. Cap. 21: Bl. 23va Dornenkrönung. Bl. 23vb Apamene nimmt Darius die Krone weg (Concubina Appomen que abstraxxit [!] regi [darüber von anderer Hand: Zorobabel] coronam de capite). Bl. 24ra Schimi verhöhnt David. Bl. 24rb Hanun, der König der Ammoniter, entehrt den Boten Davids, indem er ihm die Kleider abschneiden lässt. Cap. 22: Bl. 24va Kreuztragung. Bl. 24vb Isaak, das Opferholz tragend, folgt Abraham (mit Feuerschale). Bl. 25ra Gleichnis von den bösen Winzern: Weinbergarbeiter töten den Boten des Weinbergbesitzers. Bl. 25rb Die beiden Kundschafter tragen die große Traube. Cap. 23: Bl. 25va Christus wird ans Kreuz geschlagen. Bl. 25vb Tubal-Kaijn schmiedet Nägel und Jubal musiziert. Bl. 26ra Jesaja wird zersägt. Bl. 26rb Joab, der König von Moab, opfert seinen Sohn auf der Stadt mauer. Cap. 24: Bl. 26va Kreuzigung mit Maria, Johannes und den beiden Schächern. Bl. 26vb Traum Nebukadne zars vom großen Baum. Bl. 27ra Opfertod des Königs Codrus. Bl. 27rb Eliëser tötet den Elefanten. Cap. 25: Bl. 27va Verspottung des gekreuzigten Christus. Bl. 27vb Michal, Sauls Tochter, verspottet David. Bl. 28ra Abschalom wird von seinen Mördern verspottet. Bl. 28rb Ewil-Merodach zerstückelt den Leichnam seines Vaters Nebukadnezar. Cap. 26: Bl. 28va Kreuzabnahme. Bl. 28vb Josefs Rock wird Jakob gebracht. Bl. 29ra Adam und Eva be trauern den toten Abel (mit offenen Augen). Bl. 29rb Noomi trauert mit Rut und Orpa um ihre Söhne. Cap. 27: Bl. 29va Grablegung Christi. Bl. 29vb David folgt trauernd dem Leichnam Abners (normaler weise vier Sargträger, von den beiden hinteren nur Köpfe zu sehen). Bl. 30ra Josef wird von seinen Brüdern in den Brunnen geworfen. Bl. 30rb Jona wird ins Meer geworfen und vom Wal verschlungen. Cap. 28: Bl. 30va Christus in der Vorhölle (übliche Einteilung in vier Register nur rudimentär im oberen Bereich erhalten: zwei Register, daneben Christus auf Wolke; darunter Höllenszene). Bl. 30vb Die drei Jünglinge im Feuerofen, von einem Engel getröstet. Bl. 31ra Daniel in der Löwengrube, von einem Engel gespeist. Bl. 31rb Strauß befreit sein Junges mit Schlangenblut aus einer Glasflasche. Cap. 29: Bl. 31va Christus besiegt den Teufel. Bl. 31vb Benaja tötet einen Löwen. Bl. 32ra Simson und der Löwe. Bl. 32rb Ehud tötet Eglon (u. a. Ayoth ambidexter). Cap. 30: Bl. 32va Maria besiegt den Teufel mit den Leidenswerkzeugen. Bl. 32vb Judit tötet Holofernes. Bl. 33ra Jaël tötet Sisera. Bl. 33rb Königin Tomyris taucht das Haupt des von ihr getöteten König Cyrus in Blut. Cap. 31: Bl. 33va Höllenfahrt Christi: Jesus befreit die Seelen der Gerechten aus dem Höllenfeuer. Bl. 33vb Auszug aus Ägypten. Bl. 34ra Gott befreit Abraham aus dem Feuer (Ur in Chaldäa). Bl. 34rb Lots Flucht aus Sodom. Cap. 32: Bl. 34va Auferstehung Christi. Bl. 34vb Simson mit den Stadttoren von Gaza. Bl. 35ra Jona wird vom Wal ausgespien. Bl. 35rb Eckstein wird Schlussstein des Tempels (Lapis angularis factus in caput anguli). Cap. 33: Bl. 35va Himmelfahrt Christi. Bl. 35vb Jakobs Traum von der Himmelsleiter. Bl. 36ra Gleichnis vom guten Hirten (Ovis perdita). Bl. 36rb Himmelfahrt des Elija (Feuerwagen nicht als solcher gekenn zeichnet) (u. a. Verweis auf Parallelstelle Hoc etiam legitur de Enoch Genesis x). Cap. 34: Bl. 36va Pfingsten (Ausgießung des hl. Geistes). Bl. 36vb Turmbau zu Babel. Bl. 37ra Gott er scheint Mose (Gesetzgebung auf dem Berg Sinai). Bl. 37rb Elija füllt mit seinem Gebet die Ölkrüge der armen Witwe. Cap. 35: Bl. 37va Maria mit den Leidenswerkzeugen. Bl. 37vb Hanna sorgt sich um den abwesenden Tobias: links die Eltern Hanna und Tobit, rechts Tobias, der von seinem Begleiter Rafael weg geführt wird. Bl. 38ra Gleichnis von der verlorenen Drachme. Bl. 38rb Saul vermählt Palti und Michal, obwohl diese David liebt. Cap. 36: Bl. 38va Krönung Mariens. Bl. 38vb David bringt die Bundeslade in seinen Palast. Bl. 39ra Das Apokalyptische Weib (Mondsichel-Madonna). Bl. 39rb Salomo lässt seine Mutter Batseba zu seiner Recht en auf dem Thron Platz nehmen. Cap. 37: Bl. 39va Maria und zwei Mönche besänftigen als Fürbitter den Zorn Gottes, symbolisiert durch Pfeile, die auf die sündigen Menschen gerichtet sind. Bl. 39vb Abigajil besänftigt Davids Zorn. Bl. 40ra Die Cod. 166 189 Frau aus Tekoa besänftigt Davids Zorn. Bl. 40rb Eine kluge Frau besänftigt Joabs Zorn mit dem abgeschla genen Haupt des Scheba. Cap. 38: Bl. 40va Schutzmantel-Madonna. Bl. 40vb Tharbis verteidigt die Stadt Saba [HS]. Bl. 41ra Eine Frau tötet Abimelech mit einem Stein und rettet die Stadt Tebez. Bl. 41rb Michal rettet David vor seinen Verfolgern. Cap. 39: Bl. 41va Christus zeigt Gottvater seine Wundmale. Bl. 41vb Antipater zeigt Julius Caesar seine Wunden. Bl. 42ra Maria zeigt Christus ihre Brust (hier verhüllt) und bittet für das Volk (Maria armiger Christi). Bl. 42rb Ester vor Ahasver als Fürbitterin für die Israeliten. Cap. 40: Bl. 42va Jüngstes Gericht. Bl. 42vb Gleichnis von den zehn Talenten. Bl. 43ra Gleichnis von den fünf klugen und den fünf törichten Jungfrauen. Bl. 43rb Das Menetekel. Cap. 41: Bl. 43va Strafen der Verdammten in der Hölle. Bl. 43vb Rache an Davids Feinden. Bl. 44ra Gideon lässt die Ältesten von Sukkot schlagen. Bl. 44rb Untergang der Ägypter im Roten Meer (kein Wasser zu erkennen, sollte evtl. durch rote Kolorierung dargestellt werden). Cap. 42: Bl. 44va Maria und Christus thronend in der Mandorla. Bl. 44vb Salomo und die Königin von Saba. Bl. 45ra Gastmahl des Königs Ahasver. Bl. 45rb Gastmahl der Söhne Ijobs. Cap. 43: Die sieben Stationen des Leidens Christi. Bl. 45va Christus mit Kreuz (und Wundmalen, Sei tenwunde ist zu erkennen) erscheint einem Einsiedler. Bl. 45vb Letztes Abendmahl. Bl. 46ra Ölberg. Bl. 46rb Gefangennahme Christi (Judaskuss). Bl. 46va Christus vor Pilatus (Handwaschung). Bl. 46vb Geißelung Christi. Bl. 47ra Urteil des Pilatus und Kreuztragung Christi. Bl. 47rb Dreifigurige Kreuzigung. Cap. 44: Die sieben Schmerzen Mariens. Bl. 47va Mönch mit Schwert in der Brust. Bl. 47vb Darbringung Christi im Tempel. Bl. 48ra Flucht nach Ägypten (heilige Familie zu Fuß mit Engelserscheinung). Bl. 48rb Der zwölfjährige Christus bei den Schriftgelehrten; Mann mit wirrem Haar spuckt (?) Christus an (Phari säer?). Bl. 48va Gefangennahme Christi. Bl. 48vb Dreifigurige Kreuzigung. Bl. 49ra Grablegung Christi. Bl. 49rb Maria mit den Leidenswerkzeugen Cap. 45: Die sieben Freuden Mariens. Bl. 49va Maria erscheint einem kranken Mönch. Bl. 49vb Verkündi gung (auf dem Spruchband des Engels: Ave gracia plena). Bl. 50ra Heimsuchung. Bl. 50rb Geburt Christi. Bl. 50va Anbetung der Heiligen Drei Könige. Bl. 50vb Darbringung Christi im Tempel. Bl. 51ra Maria fin det Christus unter den Schriftgelehrten. Bl. 51rb Krönung Mariens. Illustrationen jeweils rahmenlos, häufig mit Bodenandeutungen. Etwas ungelenke Figuren, zuweilen mit unproportionierten Händen; vereinzelt ungeschickte Rückenfiguren (z. B. Bl. 45vb); bis auf ältere bzw. bär tige Männer relativ einheitliche Gesichter, häufig mit deutlich eingezeichnetem Nasenrücken. Gewänder mit kritzelig eingezeichneten Faltenbahnen, die am Boden weich umbrechen. Einfache Schattierungen durch Schraffen, häufiger durch graue Flächen (Bl. 20v Abschattierung durch Rot); insgesamt wenig Plastizität. Uneinheitliche Kolorierung in Rot (ab Bl. 44v auch Rotbraun), Grün, Ocker(-Gelb), ab Bl. 42v Blau(‑Grau); selten Inkarnat(‑Andeutungen); kolorierte und unkolorierte Abschnitte wechseln sich ab: anfangs (Bl. 3v– 25r, Cap. 1–22) sorgfältigerer Farbauftrag; Bl. 25v–33r (Cap. 23–30) bis auf etwas Rot für Blut bzw. Wun den unkoloriert; Bl. 33v–38r und 42v–43v (Cap. 31–35; 40) koloriert, aber bis auf Bl. 42v–43r mit weniger Aufwand; Bl. 38v–42r und 44r–51r (Cap. 36–39; 41–45) bis auf rotbraune Einsprengsel unkoloriert. Während die ältere Forschung, u. a. Hermann, Cod. 166 nach Österreich lokalisierte, gibt Roland, Gotik 267 Anm. 18, 271 Anm. 50, der Stilentsprechungen zum Johannesmeister aus dem Atelier des Rüdiger Schopf (um 1400, Freiburg i. B.) sieht, die Hs. nach Südwestdeutschland. Die Lokalisierung bedarf allerdings noch weiterer Überprüfung. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. etwas abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck MANUSCRIPTA in Goldrahmung, im dritten gelbes Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Die zeitgenössische Foliierung in der Mitte des oberen Blattrandes lässt – gemeinsam mit den übereinstimmenden Wasserzeichen sowie der Größe der Hs. – darauf schließen, dass Cod. 166 urspr. mit Cod. 165 vereinigt war. Gegebenenfalls wohl im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halb ledereinband. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 2 H 14 (Tinte, Bleistift) und Nr. 166/1 (Tinte) sowie Bleistift notiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 166. Bl. IIr Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben 190 Cod. 166 und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. IIr sowie Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: 1 Wilhelm II 110. – H. Tietze, Die typologischen Bilderkreise des Mittelalters in Österreich. Wiener Jahrbuch für Kunstgeschichte 2/1 (1904) 62. – Hermann 166–169, Nr. 178. – Rasmo 26 (Nr. 48) und Abb. 44–45 (ohne Nennung der Sign.). – Peintner, Buchmalerei 37–40. – Katalog ULB Tirol II 140, 207. – Peintner, Stiftsbibliothek 118–119. – Roland, Gotik 267 Anm. 18, 271 Anm. 50. (Ir–v) leer. (IIr) Stempel (s. bei G). (IIv) Memorialverse De ordine librorum Bibliae (Walther, Initia Nr. 19872). Inc.: Ut seriem menti (!) teneas textus doceam te / Est Genesis primus, post Exodus ipse secun dus … – Expl.: … Iacobus et Petrus Io[hannes] Iudas et Apokalipsis. 2 (1ra–51rb) Speculum humanae salvationis (Ed.: J. Lutz, P. Perdrizet. Mühlhausen 1907. – Steg müller RB Nr. 11765. Bloomfield Nr. 2562 und 4579). Gegenüber Edition vereinzelt Verse ausgelassen oder eingefügt, Wortstellung bisweilen leicht abweichend. (1ra–3rb) Prooemium. Tit.: Incipit prohemium istius libri qui dicitur speculum humane salva cionis. (3va–45rb) Cap. 1–42 (Ed. 4–87). (45va–47rb) De septem stationibus passionis Iesu Christi (Ed. 88–91). Tit.: Capitulum 43m. (47va–49rb) De septem tristitiis BMV (Ed. 92–95). Tit.: Capitulum 44m de septem tristiciis beate virginis. (49va–51rb) De septem gaudiis BMV (Ed. 96–99). Tit.: Capitulum 45m de septem gaudiis beate virginis. 3 (51va–52rb) Vita s. Mariae post ascensionem Christi versificata (vgl. Druck: A. Vögtlin, Vita beatae virginis Mariae et salvatoris rhythmica [Bibliothek des Litterarischen Vereins in Stuttgart 180]. Tübingen 1888, V. 6612–6741. – Walther, Initia Nr. 7529). (51va–52ra) Tit.: Qualis fuerit vita et conversacio virginis Marie. Incipit prologus operis sequen tis. – Inc.: Hec est vita virginis Marie gloriose / Summi Dei filii matris generose … Maria sibi regulam statuit vivendi / et sub quodam ordine Deo serviendi … – Expl.: … Sic per dies singulos Maria faciebat / Quibus in hoc seculo vivens permanebat. (52ra–rb) Anschließend Mariengebet in Versen. Tit.: Oracio ad beatam virginem. – Inc.: O regi na virginum fac tecum gratulari / Quos hic in exilio vides tribulari … – Expl.: … Gaudentesque perducere ad celi regna Amen. Parallelüberlieferung: u. a. Graz, UB, Hs. 241, Bl. 126v und Hs. 1638, Bl. 69r. (52rb) Schlussschrift (rot): Explicit speculum humane salvacionis / Quod docet nos venire ad locum eterne visionis. 4 (52va–56ra) Auszüge aus ‹Eusebius Caesariensis, interprete Rufino Aquileiensi›: Ecclesiasticae Historiae (Druck: u. a. Basel 1559. Ed.: E. Schwartz, Th. Mommsen, Eusebius Werke II: Historia ecclesiastica [GCS 9/1 und 2]. Leipzig 1903 und 1908). Tit.: Nota de destinccione (!) gentis Iudeorum. – Inc.: Quanta Iudeis acciderint gravia post post (!) temeritatem crucis in Christum scribit magister in Ecclesiastica Hystoria libro 2 co 6 dicens Primo quidem temporibus Tyberii refert in urbe Roma … – Expl.: … sic deterriti Iudei atque gentiles locum simul et inaniter cepta relinquere. (52va–53rb) Lib. 2. (52va) Cap. 6. – (52vb) Cap. 7 und 11. – (53ra) Cap. 19 und 20. – (53rb) Cap. 21 (leicht gekürzt). (53rb–55va) Lib. 3. (53rb) Cap. 5. – (53va) Cap. 6. – (54vb) Cap. 7. – (55ra) Cap. 8. (55va–55vb) Lib. 4, cap. 6. Cod. 167 191 (55vb–56ra) Lib. 10 (= Rufinus Aquileiensis, Lib. 1), cap. 38–40 (Druck auch PL 21, 505A–506C, hier cap. 37–39. – Vgl. CPL Nr. 198k). (56rb–vb) nur Rahmung. 5 (57ra) ‹Sulpicius Severus, Iohannes Cassianus›: De vitis patrum, Lib. 4 = Excerpta, cap. 45, unvollständig, durchgestrichen (PL 73, 842A). Inc.: [Q]uid eciam de illo monacho commemorem cuius nomen quia adhuc superest … – Expl. mut.: … pariter in eodem monasterio commanebat [ (57rb–58vb) nur Rahmung. (Ir–v) leer. A. P. / U. S. Cod. 167 ADAM DE ALDERSBACH Pap. I, 72, I* Bl. 290 × 205. Süddeutscher Raum, 1419. B: Pap. fleckig (Wasserschaden?), Bl. 39 und 40 löchrig. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Verwandte Wasserzeichen: u. a. Wien, Schottenstift, Archiv, Hs. 329, T. I (dat. 1417); Klosterneuburg, Stiftsbibl., CCl 680, T. I (dat. 1419); Innsbruck, ULB Tirol, Cod. 546. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + VII14 + 4.VI62 + V72 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am Ende der Lagen 4 und 5 Kustode in Rot in arabischer Ziffer, am Ende der Lage 4 stark beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 215/225 × 145/150. Zwei Spalten, von Tintenlinien gerahmt, ab Bl. 39 nur mehr seitlich be grenzt, zu 44–51 Zeilen. Verszeilen mit größerem Abstand (ca. drei Zeilen des begleitenden Kommentars). Bastarda, dat. 1419 (Bl. 71va). Schreiber: Nicolaus Frechen. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen und Überschriften. Drei- bis zehnzeilige rote Lombarden, tw. mit Punktverdickungen. Bl. 1ra fünfzeilige rote Initiale mit ausgesparten Vierblatt blüten. Bl. 31v (ganzseitig) Arbor consanguinitatis in schwarzer und grünlich-ockerfarbener Pinselzeich nung; Baumschema entspringt aus einem Kranz von Akanthusblättern, Medaillons nicht beschriftet. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. abgerieben und beschädigt. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten RI TUALE in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 1r Besitzvermerk Anfang 16. Jh. Magister Nicolaus Leopoldus ex Insprugga XXX (vgl. Cod. 579; 1511 Lehrer an der Pfarrschule von St. Jakob, Innsbruck [W. Steiner, Die Lateinschule der St. Jakobspfarrkirche in Innsbruck, in: K. Drexel, M. Fink (Hrsg.), Musikgeschichte Tirols I: Von den Anfängen bis zur Frühen Neuzeit (Schlern-Schriften 315). Innsbruck 2001, 404, 501], empfängt 1513 in Brixen die niederen Weihen und erhält 1515 Kanonikat, gest. 1535 [K. Wolfsgruber, Das Brixner Domkapitel in seiner persönlichen Zusammensetzung in der Neuzeit 1500–1803 (Schlern‑Schriften 80). Innsbruck 1951, 42 und 170; W. Senn, Musik und Theater am Hof zu Innsbruck. Geschichte der Hofkapelle vom 15. Jahrhundert bis zu deren Auf hebung im Jahre 1748. Innsbruck 1954, 13 und 46]). Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 15 (Tinte, Bleistift) und N. 167 (Tinte) sowie Bleistift notiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 167. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. 192 L: 1 Cod. 167 Wilhelm II 108. – Katalog ULB Tirol II 140, 207. – Katalog ULB Tirol II, Beiheft, Taf. 54. – Stefani 104. – Peintner, Stiftsbibliothek 121. – Baroffio 324, Nr. 16713. (Ir–v) leer. (1ra–71vb) ‹Adam de Aldersbach›: Summula pauperum mit Kommentar und Interlinearglossen (bis Bl. 66vb) (Druck der Verse: GW 212–217. – Walther, Initia Nr. 9117. Kaeppeli Nr. 3. Bloom field Nr. 2668 und 5852. Kommentar nicht übereinstimmend). (1ra–b) Prolog. Inc.: Secundum dictum Augustini in domo Dei id est in ecclesia quatuor sunt necessaria scilicet grammatica musica computus et ius cananicum (!). Gramatica ad scripturas sacras intelligendum musica ad laudes Dei decantandum … – Expl.: … Civile dicitur alio nomi ne imperiale et illud traditur in quattuor libris legum scilicet ius canonicum est ius regulare et traditur in duobus libris principaliter scilicet in Decretalibus et in libro Decretorum et in aliis. (1rb–71ra) Text. Inc.: In summis festis (interlinear: in diebus magne festivitatis) ad missam (inter linear: id est divinum officium) dicitur (interlinear: id est legitur) una. Tantum (interlinear: id est solummodo) collecta nisi sit (interlinear: semper talis festivitatibus) coniuncta (interlinear: ap probata) diei (interlinear: semper alicuius sancti). (Forts. 2rb) In defunctorum (interlinear: id est mortuorum) collectam (interlinear: id est talem oracionem) dicere (interlinear: id est legere vel interponere) missa de sanctis poteris (interlinear: id est valebis) sed (interlinear: id est tamen) illa penultima fiat (interlinear: semper collecta id est penes id est erit ultimam) … – Expl.: … virgo si rapitur a pluribus et violenter unus noscat eam … in summis festis que quatuor esse videtur (!) corpora non aliqua defunctorum tumulantur. (1rb–71vb) Kommentar. Inc.: Hic iam est pars executiva in qua prosequitur intentum volens depellere ignoranciam deposicionis circa quid sciendum quod duplex est ignorancia scilicet ignorancia abnegacionis et ignorancia deposicionis … – Expl.: … In ista parte magister ponit alia documenta circa confessionem et primo facit hoc de raptu virginis … sed communiter dicta sunt festum ascensionis et festum cordis Christi et festum omnium sanctorum et patrocinium et quodlibet alius festum beate virginis Marie et sic est finis Deo laus omnipotenti et matri eius gloriosissime virginis Marie. Laus summo genitori detur atque eius genitrici Deo gracias. (31v) ganzseitig Arbor consanguinitatis, nicht beschriftet. (71va) Schlussschrift mit Datierung und Schreibernennung: Anno ab incarnacione domini mille simo quarto decimo (!) et decimo nono finitus est liber iste per manus Nicolay Frechen in octava Agnetis virginis hora decima diei ante festum purificacionis Marie virginis etc. (1. Februar 1419). Lit.: Schulte GQ II 427–428; F. Valls Taberner, La „Summula Pauperum“ de Adam de Alders bach (Span. Forschungen der Görresgesellschaft I/7). Münster 1938, 75–83. 2 (71va–b) Exzerpte aus Augustinus, Gregorius Magnus und Beda Venerabilis. 1) Tit.: Augustinus. – Inc.: In illo die quo videtur corpus Christi victus necessarius retribuitur lenia colloquia dimittantur … – Expl.: … a Deo remunerantur etc. 2) Tit.: Gregorius. – Inc.: O quam magnum donum et mirificum bonum de Dei misericordia quia tanta est dignitas … – Expl.: … unius anime a penis purgatorii liberacio etc. 3) Tit.: Beda. – Inc.: Sacerdos qui sine mortalibus est peccatis et bono proposito si non celebrat missam cum possit et habet copiam celebrandi … – Expl.: … et pro hoc respondebit Deo in no vissimo die. 3 (71vb) Versus de regimine sacerdotis (Walther, Initia Nr. 14519). 16 Hexameter. Inc.: Presbiter attende capias hec dogmata mente / hec tibi norma datur per quam tua vita regatur … – Expl.: … vivere sine mori (!) confert qui porrigit ori. 4 (71vb) De Transsubstantiatione (Walther, Initia Nr. 3214). 14 Hexameter. Tit.: Augustinus. – Inc.: Constat in altari carnem de pane creari / illa caro Deus est qui dubitat reus est … – Expl.: … errores hereses laicorum quando reiecit. Cod. 168 5 193 (72r) ‹Ps.-Bonaventura›: Tabula a celebrantibus missam consideranda (Druck: GW M14225. – Distelbrink Nr. 225). Mit acht schematischen Darstellungen. Inc.: Intencionis discussio – ne propter vanam gloriam … – Expl.: … causa excellentis / excellen tioris anime / excellentissime divinitatis Christi. Darunter Notiz: Tangere qui gaudes meretricem qualiter audes / palmis pollutis regem tractare salutis (Walther, Proverbia Nr. 31040). (72v) nur Rahmung. (Ir–v) leer. P. A. Cod. 168 ARISTOTELES Pap. I, 46, I Bl. 290 × 220. Wien, 9. Jz. 14. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Göttweig, Stiftsbibl., Cod. 70 (rot) (dat. 1384); Zwettl, Stiftsbibl., Cod. 224 (dat. 1389). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 3.VI36 + V46 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Erstes und letztes Bl. des Quinternio von den übrigen Bl. etwas abweichend, unsicher, ob zwei einzelne Bl. oder ein Doppelbl. Lagenreklamanten. Am oberen Blattrand rechts zeitgenössische Zählung der Bücher in arabischen Ziffern, tw. durchgestrichen und neu geschrieben. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Text von Kommentar umgeben. Schriftraum Haupttext 160 × 95/110, von Tintenlinien gerahmt, zu 27–33 Zeilen. Schriftraum Kommentarfelder stark wechselnd, von Tintenlinien gerahmt (Bl. 1–36 einfach, Bl. 37r– 46v tw. zweifach). Äußeres Kommentarfeld tw. beschnitten (Textverlust). Kursive Bastarda von mehreren einander abwechselnden Händen. Schreiberwechsel Bl. 2v, 3v, 5v, 10v, 12v/13r, 16v, 17r (die letzten drei Zeilen gleiche Hand wie die dazugehörige Glosse), 18v, 28r/v, 34v/35r, 35r/v, 36v/37r, 38r–39r Forts. von Bl. 36v. Bl. 35r Text in kalligraphischer Bastarda. Weitere Schreiberwechsel 39r, 40r, 41r/v, 41v/42r. Glos sen weitgehend von einer Hand. Vgl. auch Cod. 163, T. I. A: Rote Auszeichnungsstriche, Unterstreichungen, Rahmungen sowie schwarze, rot konturierte Paragraph zeichen. Zwei- und dreizeilige rote und schwarze Lombarden, tw. mit rotem (Schraffur-)Dekor, Bl. 1r, 9r, 24v vier- und fünfzeilige rote Lombarden, Bl. 1r mit schwarzen gegenständigen Halbpalmetten im Binnen feld. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. PHYSICA in Goldrahmung, im dritten Signatur schild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistift notiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Innsbruck, ULB Tirol, Cod. 157, Bl. Ir gehört Cod. 168 zu einer Gruppe von urspr. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wur den. Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 157: Aristoteles: Ethica Nicomachea und Oeconomica. heute nicht mehr nachweisbar: Aristoteles: Politica. Cod. 159: Albertus de Saxonia: Kommentar zu Cod. 157. Cod. 168: Aristoteles: Physica, Lib. 1–5. Cod. 163: Aristoteles: Physica, Lib. 6–8 und De anima. Cod. 181: Alanus ab Insulis: Summa de arte praedicatoria. Cod. 164: Anshelmus Havelburgensis. Anticimenon. 194 Cod. 168 Von diesen sieben Hs. ist eine nicht mehr erhalten, drei befinden sich heute an der ULB Tirol in Innsbruck (Cod. 157, 159 und 164), drei werden in der Stiftsbibl. Neustift (Cod. 163, 168 und 181) aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 2 H 16 (Tinte, radiert), II 2 H 16 (Bleistift) und Nr. 168 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 168. Bl. 2r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abge geben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 2r Stempel der Soprin tendenza Tridentina 20. Jh. L: Lacombe II 1024 Nr. 1496. – Neuhauser, Neustift 91–95. – Katalog ULB Tirol II 140, 207. – W. Neuhauser, Buchgeschichtliche Beziehungen zwischen dem Wiener Raum und Tirol im Mittelalter. Jahrbuch für Lan deskunde von Niederösterreich N. F. 63/64 (1997/98) 57 (auch in: Neuhauser, In libris 405–424). (Ir–v) leer. (1r) am oberen Blattrand Schreibervers, stark beschnitten: Assit ad inceptum sancta Maria meum. (1r–46v) Aristoteles: Physica, Translatio nova durch Guilelmus de Moerbeke, Lib. 1–5 (Druck: u. a. GW 2336. Ed.: Aristoteles Latinus VII: Physica, Fasc. 1: Translatio vetus, ed. F. Bossier, J. Brams, rev. ed. A. Mansion. Leiden, New York 1990. – Lacombe I 52, Nr. 17, I 126–127, Nr. 17 und II 1024, Nr. 1496. Thorndike–Kibre 1296). Mit umfangreichem Kommentar. Tit.: Textus VIII librorum Phisicorum. – Inc.: Quoniam quidem scire et intelligere contingit circa omnes sciencias quarum principia sunt aut cause aut elementa … – Expl. mut. (Lib. 5, cap. 6, knapp vor Schluss): … et de motu quidem et quoete (!) quomodo utique unus [ (1r–8v) Lib. 1. – (9r–17r) Lib. 2. – (17r–24r) Lib. 3. – (24v–39r) Lib. 4. Bl. 36v schließt propter parvita tem, Bl. 37r (Forts. von anderer Hand): et perpetuitatem. Mutatio autem omnis a natura … bis inpossibile est et [ Bl. 37v leer, Bl. 38r (von gleicher Hand von Bl. 36v): Mutatio autem omnis a natura … – (39r– 46v) Lib. 5. Bl. 45r–v leer. Text von Bl. 44v setzt auf Bl. 46r unmittelbar und ohne Textverlust fort. (1r–46v) Kommentar. (1r) Einleitender Kommentar zum gesamten Werk. Inc.: Dubitatur utrum sciencia naturalis est sciencia de omnibus rebus … – Expl.: … et que quando cum ad formationem. (1r–8v) Zu Lib. 1. Inc.: Quoniam quidem. Iste liber intytulatur liber Phisicorum Aristotelis filii Nicometi Grecorum sapientissimi … – Expl.: … pateant declaranda. Et sic est finis primi libri Phisicorum etc. Deo gratias Amen etc. (9r–17r) Zu Lib. 2. Inc.: Eorum que. Iste est liber secundus Phisicorum in quo Aristoteles determinat de p (!) principiis doctrine … – Expl.: … et sic est finis secundi de quo sit benedictus Ihesus Marie filius Amen etc. (17r–24r) Zu Lib. 3. Inc.: Quoniam autem natura. Hic incipit 3us liber Phisicorum (gestrichen: Et continet) in quo Aristoteles determinat de passionibus … – Expl.: … Et sic est finis Aristotelis 3ii libri de quo sit benedictus Ihesus Marie filius in secula seculorum Amen Amen etc. (24v–39r) Zu Lib. 4. Inc.: Similiter autem. Hic est quartus liber Phisicorum in quo Aristoteles determinat de passionibus … – Expl.: … et anima in 10 casibus. (39r–46v) Zu Lib. 5. Inc.: Transmutatur autem. In ista parte Aristoteles ponit 5m librum Physicorum in quo Aristoteles determinat de divisione motus … – Expl.: … solvit ut presens fuit soluta. Bl. 46v (am unteren Blattrand) Reklamante zu Lib. 6: Si autem est continuum. Forts. (Lib. 6–8) in Cod. 163, Bl. 1r–29r. Lit.: Grabmann, Aristoteles 170–174. (Ir–v) leer. P. K. / W. N. Cod. 171 195 Cod. 171 THOMAS CANTIMPRATENSIS. THEOLOGISCHE TRAKTATE Perg. I, 91, I Bl. 285 × 210. Süddeutscher Raum, Ende 13./Anfang 14. Jh. B: Zwei Teile: I (I, 1–61), II (62–91, I). Am oberen Blattrand durchgehende Foliierung 19. Jh. durch die ULB Tirol. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck DE NATURA LIB. in Gold rahmung, im dritten Signaturschild (s. bei G). Schnitt rot gefärbt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neu stift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signatur der ULB Tirol II`1 […] (Tinte, radiert), II 2 H 19 (Bleistift) und Nr. 171/ (1) (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 171/1. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 142, 207. I (I, 1–61) B: Perg. tw. löchrig, Bl. 19 Riss mit rotem Zwirn genäht. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + V10 + (V–4)16 + 4.V56 + (V–5)61. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. In der zweiten Lage zwei Doppelbl. verloren (Textverlust). Nach Bl. 61 fünf leere, bis auf das letzte unlinierte Blätter am Ende der Lage weggeschnitten. Kustoden in römischen Zahlen (IIIus–VIIIus) am Ende der ersten sechs Lagen. Zeit genössische Foliierung in arabischen Ziffern am oberen Blattrand, beginnend mit 21; Fehler in der Zählung: Bl. 40v fehlerhafte Foliierung 55, korr. aus 56, auf Bl. 41r fehlerhafte Foliierung 56 neben der richtigen 65, auf Bl. 42r Foliierung 66 korr. aus 56. Textverlust am Anfang des Buchblocks (zwei Lagen). S: Schriftraum 205 × 120. Zwei Spalten, von Tintenlinien gerahmt, zu 40 Zeilen auf Tintenlinierung. Zirkel stiche. Textualis. Gleicher Schreiber wie T. II. Vereinzelt Korrekturen des Schreibers und kurze Anmerkungen von anderen, späteren Händen am Blattrand. Bl. 61vb acht Zeilen (Schlussschrift?) radiert. A: Rote Paragraphzeichen und Überschriften. Zweizeilige rote Lombarden. 1 (Ir–v) leer. (1ra–61vb) ‹Thomas Cantimpratensis›: Liber de natura rerum, unvollständig (Ed. H. Boese. Ber lin, New York 1973, 13–377. – Thorndike–Kibre 1026). Einzelne Kapitel gegenüber der Ed. tw. gekürzt oder erweitert, Kapiteleinteilung bisweilen abweichend, ganze Kapitel ausgelassen oder hinzugefügt. Enthält: (1ra–3rb) Lib. 4. (1ra) Cap. 80. Inc. mut.: ca]ventes in secretis pariunt et occultant. Femine onagrorum … – (1ra) Cap. 82. – Cap. 85. – (1va) Cap. 86. – Cap. 87. – (1vb) Cap. 90. – Cap. 93. – Cap. 94. – (2ra) Cap. 96. – (2rb) Cap. 98. – (2va) Cap. 101. – Cap. 103. – Cap. 97. – (2vb) Cap. 104. – (3ra) Cap. 105. – (3rb) Cap. 108. (3rb–13va) Lib. 5. (3rb) Cap. 1. – (4ra) Cap. 2. – (4va) Cap. 4. – Cap. 6. – (4vb) Cap. 8. – Cap. 9. – (5ra) Cap. 10. – (5rb) Cap. 11. – Cap. 12. – Cap. 14. – (5va) Cap. 15. – (5vb) Cap. 18. – Cap. 24. – (6ra) Cap. 26. – Cap. 27. – Cap. 28. – (6rb) Cap. 30. – (6va) Cap. 31. – Cap. 32. – (6vb) Cap. 34. – (7ra) Cap. 35. – Cap. 36. – (7va) Cap. 38. – (7vb) Cap. 39. – (8ra) Cap. 40. – Cap. 41. – (8rb) Cap. 44. – Cap. 45. (8vb) Cap. 51. – (9ra) Cap. 52. – (9rb) Cap. 53. – Cap. 55. – (9va) Cap. 57. – (9vb) 196 Cod. 171 Cap. 58. – (10va) Cap. 59. – (10vb) Cap. 60. – (11ra) Cap. 61. – Cap. 62. – (11rb) Cap. 63. – Cap. 66. – (11va) Cap. 67. – (11vb) Cap. 73. – Cap. 74. – Cap. 76. – Cap. 79. Expl. mut.: … domesticis rebus insidiatur. Accipitri [ – (12ra) Cap. 106. Inc. mut.: ] pullorum evolancium inbecilles … – Cap. 108. – Cap. 109. – (12rb) Cap. 110. – (12va) Cap. 111. – Cap. 112. – (12vb) Cap. 114. – Cap. 113. – Cap. 116. – (13ra) Cap. 118. – Cap. 119. – (13rb) Cap. 120. (13va–15va) Lib. 6. (13va) Cap. 2. – (13vb) Cap. 3. – Cap. 5. – Cap. 7. – (14ra) Cap. 10. – Cap. 12. – (14rb) Cap. 13. – Cap. 15. – Cap. 16. – (14va) Cap. 20. – Cap. 23. – (14vb) Cap. 27. – Cap. 30. – Cap. 33. – (15ra) Cap. 34. – Cap. 36. – Cap. 46. – (15rb) Cap. 47. – Cap. 48. – (15va) Cap. 53. (15va–16va) Lib. 7. (15va) Cap. 1. Expl. mut: … aliqui non tangunt vas quo venantur nisi sit recens. Femine pis ci[um – (16ra) Cap. 52. Inc. mut.: ne]mo propheta acceptus est in patria sua … – Cap. 57. – Cap. 59. – Cap. 64. – (16rb) Cap. 70. – Cap. 72. – (16va) Cap. 77. – Cap. 83. – Cap. 88. (16va–21vb) Lib. 8. (16va) Cap. 1. – (17rb) Cap. 2. – Cap. 3. – (17va) Cap. 4. – (17vb) Cap. 5. – (18ra) Cap. 6. – Cap. 9. – (18rb) Cap. 12. – (18rb) Cap. 13. – Cap. 14. – Cap. 15. – (18va) Cap. 16. – (19ra) Cap. 17. – Cap. 20. – (19rb) Cap. 21. – Cap. 23. – Cap. 25. – (19va) Cap. 26. – (19vb) Cap. 27. – Cap. 29. – Cap. 30. – (20rb) Cap. 31. – Cap. 32. – Cap. 33. – (20va) Cap. 34. – De serpente vulgari. – Cap. 35. – Cap. 36. – Cap. 37. – (20vb) Cap. 38. – Cap. 40. – (21ra) Cap. 41. – Cap. 42. – Cap. 43. – (21rb) Cap. 45. (21vb–26rb) Lib. 9. (21vb) Cap. 1. – Cap. 2. – (·23rb) Cap. 3. – Cap. 5. – (23va) Cap. 7. – (23vb) Cap. 9. – Cap. 11. – Cap. 12. – (24ra) Cap. 13. – Cap. 14. – (24rb) Cap. 15. – Cap. 16. – Cap. 19. – (24va) Cap. 21. – Cap. 22. – (24vb) Cap. 24. – Cap. 25. – (25ra) Cap. 28. – (25rb) Cap. 33. – Cap. 34. – Cap. 35. – (25va) Cap. 43. – Cap. 45. – (25vb) Cap. 44. – Cap. 40. – Cap. 46. – Cap. 48. – (26ra) Cap. 47. – Cap. 49. – Cap. 51. – Cap. 52. – Cap. 53. (26rb–32ra) Lib. 10. (26rb) Cap. 5. – Cap. 6. – (26va) Cap. 7. – Cap. 8. – Cap. 12. – (26vb) Cap. 13. – Cap. 15. – (27ra) De cyconia. – Cap. 16. – (27rb) Cap. 18. – (27va) Cap. 19. – Cap. 20. – Cap. 21. – (27vb) Cap. 22. – Cap. 23. – (28ra) Cap. 25. – Cap. 26. – Cap. 24. – (28rb) Cap. 28. – Cap. 29. – (28va) De mirica. – Cap. 31. – (28vb) De mepsilo (!). – Cap. 33. – Cap. 32. – Cap. 34. – (29rb) Cap. 35. – Cap. 36. – (29va) Cap. 37. – Cap. 38. – Cap. 39. – (29vb) De pyro. – (30ra) De pruna. – Cap. 40. – Cap. 41. – Cap. 42. – (30va) Cap. 43. – (30vb) Cap. 45. – De sicomoro. – De sambuco. – De sparago. – Cap. 46. – (31ra) Cap. 47. – Cap. 48. – Cap. 51. – (32ra) Cap. 52. (32ra–36rb) Lib. 11. (32ra) Cap. 2. – (32rb) Cap. 3. – Cap. 4. – (32va) Cap. 6. – Cap. 8. – (33rb) Cap. 9. – (33va) Cap. 10. – (33vb) Cap. 11. – Cap. 12. – (34ra) Cap. 13. – Cap. 14. – (34rb) Cap. 15. – Cap. 16. – Cap. 17. – (34va) Cap. 18. – Cap. 20. – (34vb) Cap. 21. – Cap. 22. – (35ra) Cap. 23. – Cap. 24. – Cap. 25. – (35rb) Cap. 26. – Cap. 27. – (35va) Cap. 28. – Cap. 29. – (35vb) Cap. 30. – Cap. 31. – Cap. 32. – (36ra) Cap. 33. (36rb–41ra) Lib. 12. (36rb) Cap. 1. – (36va) Cap. 2. – (36vb) Cap. 3. – Cap. 4. – De aristologia. – (37ra) De arthe mesia. – De alleo. – De alchana. – De anisio. – (37rb) De bethonica. – De basilisco. – De celi donia. – De cynoglosa. – De cyclamine. – (37va) Cap. 5. – Cap. 6. – (37vb) De caulibus. – De cicorea. – De canna melis. – Cap. 7. – Cap. 8. – (38ra) Cap. 9. – Cap. 10. – (38rb) Cap. 11. – De eruca. – De eleboro. – Cap. 12. – (38va) De fabis. – (38vb) De grano frumenti. – De galbano. – Cod. 171 197 Cap. 13. – (39ra) De ysopo. – De barba iovis. – De latuca. – De lylio. – Cap. 14. – (39rb) Cap. 15. – (39va) Cap. 17. – Cap. 18. – Cap. 19. – (39vb) Cap. 20. – Cap. 21. – Cap. 23. – (40ra) Cap. 24. – (40rb) De rafano. – Item de ruta. – De syler. – Cap. 25. – (40va) Cap. 26. – Cap. 27. – Cap. 28. – De squilla. – De tapsia. – (40vb) De verbena. – De vicia. – Cap. 29. – Cap. 30. – (41ra) Cap. 31. – Cap. 32. (41ra–51ra) Lib. 14. (41ra) Cap. 1. – (41vb) Cap. 2. – Cap. 3. – (42ra) Cap. 4. – (42rb) Cap. 5. – (42va) Cap. 6. – Cap. 7. – Cap. 8. – Cap. 9. – De amandino. – Cap. 10. – (42vb) Cap. 11. – Cap. 12. – (43ra) Cap. 13. – (43rb) Cap. 14. – Cap. 15. – (43va) Cap. 16. – Cap. 17. – (43vb) Cap. 18. – Cap. 19. – Cap. 20. – Cap. 21. – (44ra) Cap. 22. – Cap. 23. – De celonte. – De crisopasion. – De core. – (44rb) De demonio. – Cap. 24. – Cap. 25. – (44va) Cap. 26. – De egolito. – Cap. 27. – Cap. 28. – (44vb) Cap. 29. – De episcute. – (45ra) De exacolito. – Cap. 30. – Cap. 31. – Cap. 32. – (45rb) Cap. 33. – Cap. 34. – Cap. 35. – Cap. 36. – (45va) De gerachirea. – Cap. 37. – Cap. 38. – (45vb) Cap. 41. – Cap. 42. – Cap. 44. – (46ra) De lagapis. – Cap. 45. – (46rb) Cap. 46. – Cap. 48. – De muro. – De nichamar. – De noche. – (46va) Cap. 49. – (46vb) Cap. 50. – Cap. 51. – Cap. 52. – (47ra) Cap. 53. – Cap. 54. – Cap. 55. – Cap. 56. – (47rb) De peanite. – De quirin. – De quini dros. – Cap. 57. – (47vb) Cap. 58. – (48ra) Cap. 59. – Cap. 60. – (48rb) Cap. 61. – Cap. 62. – Cap. 63. – Cap. 64. – (48va) Cap. 66. – Cap. 65. – Cap. 67. – (48vb) Cap. 68. – De terebole. – De tervice. – De tercillo. – De zunicho. – (49ra) De zignites. – Cap. 69. – (49vb) Cap. 70. – (50vb) Cap. 71. (51ra–52va) Lib. 15. (51ra) De metallis. – Cap. 1. – (51va) Cap. 2. – (51vb) Cap. 3. – (52ra) Cap. 4. – Cap. 5. – (52rb) Cap. 6. – (52va) Cap. 7. (52va–53rb) Lib. 13. (52va) Cap. 6. – (52vb) Cap. 7. – Cap. 10. – Cap. 11. – Cap. 12. – Cap. 13. – Cap. 14. – Cap. 15. – (53ra) Cap. 16. – Cap. 18. – Cap. 19. – Cap. 20. – Cap. 21. – Cap. 24. – Cap. 27. – (53rb) Cap. 28. – Cap. 29. – Cap. 30. (53rb–61rb) Lib. 1. (53rb) Cap. 1. – (53va) Cap. 2. – (53vb) Cap. 3. – (54ra) Cap. 4. – (54rb) Cap. 5. – Cap. 6. – (54va) Cap. 7. – (54vb) Cap. 8. – Cap. 9. – Cap. 10. – (55ra) Cap. 11. – Cap. 12. – (55rb) Cap. 13. – (55va) Cap. 14. – (55vb) Cap. 15. – Cap. 16. – Cap. 17. – (56ra) Cap. 18. – Cap. 19. – Cap. 20. – (56rb) Cap. 21. – Cap. 22. – (56va) Cap. 23. – Cap. 24. – Cap. 25. – (56vb) Cap. 26. – Cap. 27. – (57ra) Cap. 35. – (57va) Cap. 36. – (57vb) Cap. 37. – Cap. 38. – (58ra) Cap. 39. – Cap. 41. – (58rb) Cap. 42. – Cap. 43. – Cap. 45. – (58va) Cap. 46. – Cap. 47. – (59rb) Cap. 48. – Cap. 49. – (59va) Cap. 50. – (60ra) Cap. 51. – Cap. 52. – (60rb) Cap. 53. – Cap. 55. – (60va) Cap. 56. – Cap. 57. – (60vb) Cap. 59. – Cap. 60. – (61ra) Cap. 62. – Cap. 63. – Cap. 65. Lit.: H. Boese, Zur Textüberlieferung von Thomas Cantimpratensis’ Liber de natura rerum. Archivum Fratrum Predicatorum 39 (1969) 53–68. (61v) leer. II (62–91, I) B: Perg. tw. löchrig, Lagen: 3.V91 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Kustoden in römischen Zahlen am Anfang und Ende der letzten zwei Lagen (IIus–IIIus). Textverlust am Ende des Buchblocks (mindestens eine Lage). Am oberen Rand der Bl. 75v, 76v–77r, 78v– 79v, 89v–91v Minuskel c. S: Schriftraum 205 × 120. Zwei Spalten, von Tintenlinien gerahmt, zu 40 Zeilen auf Tintenlinierung. Zirkel stiche, großteils weggeschnitten. Textualis. Gleicher Schreiber wie T. I, etwas kleinerer Schriftgrad. Ver 198 Cod. 173 einzelt Korrekturen des Schreibers und kurze Anmerkungen von anderen, späteren Händen am Blattrand. Schreiberspruch Bl. 62r sowie spätere Ergänzung Bl. 65v Textualis cursiva. A: Rote Paragraphzeichen und Überschriften. Bl. 62r–87v rote Marginalien. Bl. 62ra–88rb zweizeilige rote Lombarden. Bl. 63ra–91vb rote lombardenförmige Satzmajuskeln. Repräsentanten am Blattrand in Schwarz oder Rot. 2 (62ra–88rb) De comparationibus BMV (Stegmüller RB Nr. 8879). Tit. (am Blattrand): De duodecim proprietatibus. – Tit.: In isto libello continentur proprietates diversarum creaturarum similitudinarie convenientes virtutibus matris creatoris. – Inc.: Beata virgo comparatur celo. Celum enim dicitur quasi gaza eliorum id est solis domus, quia quasi in domo propria … – Expl.: … tuam precibus et meritis illius terre cui benedixisti. 3 (88rb–91vb) Tractatus super Missus est (Exzerpte aus Bernardus Claraevallensis), unvollständig. Tit.: Bernhardus super ewangelium. – Inc.: Missus est Gabriel angelus (Lc 1,26). Vehemencia desiderii, fervore devocionis, puritate oracionis fontem attingit pietatis … – Expl. mut.: … qui ipsam invocatam in necessitate non senserit affuisse et post in periculis et [ Parallelüberlieferung: Heiligenkreuz, Stiftsbibl., Cod. 112, Bl. 74r. (Ir–v) leer. L. Š. Cod. 173 MAURUS SERVIUS HONORATUS Pap./Perg. I, 172, I Bl. 260 × 215. Italien, 2. Drittel 15. Jh. B: Pap. und Perg. gemischt: In Lage 1 innerstes Doppelbl. Perg., in Lage 5, 6, 8 und 9 jeweils äußerstes und innerstes Doppelbl. Perg. Haar- und Fleischseite deutlich unterscheidbar. Pap. und Perg. tw. fleckig. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: (I–1)I (Vor satzbl., Pap. 18. Jh.) + VII14 + 3.VI50 + VIII66 + IX84 + (VIII–1)99 + VIII115 + X135 + VIII151 + (X+1)172 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufge klebt. Gegenbl. zu Bl. 99 herausgeschnitten (Textverlust). Bl. 172 eingehängtes Einzelbl. (Perg.). Bl. 172 klebte wohl als Spiegel auf dem HD des urspr. Einbandes. Zw. Bl. 39 und 40 sowie zw. Bl. 111I und 113 jeweils ein kleines Einzelbl. (200 × 60 bzw. 180 × 75) eingehängt. Zw. Bl. 135 und 136 fehlt mindestens eine Lage (Textverlust). Ab Lage 4 Reklamanten erhalten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 111 doppelt angeführt, 112 ausgelassen. In Abschnitt 3 am oberen Blattrand Buchzählung: Bl. 27v–33r und 39r–40r am oberen Blattrand mittig in Rot I für Buch 1 der Aeneis, ab Buch 2 mittig in arabischen Ziffern in Schwarz, vielfach be- bzw. weggeschnitten, und rechts in Rot, auf 8 folgt 10 (Buch 9 fehlt). S: Schriftraum 180/200 × 140/145. Zwei Spalten, von tw. sehr feinen Bleistift-, tw. Blindlinien gerahmt, zu 35–37 Zeilen auf Bleistift- bzw. Blindlinierung. Zirkelstiche. Humanistische Minuskel von drei verschiede nen Händen: 1) Bl. 1ra–120va; 2) Bl. 120vb–135vb (sehr klein, kalligraphisch); 3) 136ra–170va (flüchtiger). Am Blattrand Notizen vorwiegend von zeitgenössischer, vereinzelt von späterer Hand. A: Bl. 1ra–3rb rote Auszeichnungsstriche, Randmarkierungen in Form von vereinfachtem Perlenbesatz, seitli che Hervorhebungen, Marginalien und Schlussschrift. Zeigehände (z. B. Bl. 17ra, 18rb). Bl. 27ra–33rb und 39ra–40rb rote Auszeichnungsstriche und Unterstreichungen. In der gesamten Hs. Platz für mehrzeilige Initialen freigelassen, nicht ausgeführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. SERVY EXPOSITIO IN LIBROS ENEIDOS in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. Cod. 173 199 G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Auf hebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 H 21 (Bleistift) und N. 173 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signatur schild der ULB Tirol 173. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 136. – Neuhauser, Neustift 84. – Katalog ULB Tirol II 143, 208. – Peintner, Stiftsbibliothek 122. (Ir–v) leer. (1ra–172va) Maurus Servius Honoratus: Commentarii (Ed. G. Thilo, H. Hagen. Lipsiae 1881–1902), mit ergänzenden Notizen von späteren Händen, u. a. Zitate. 1 (1ra–3rb) Commentarius in Vergilii Bucolica, unvollständig: Lib. 9,5–Lib. 10 (Thilo III 1, 109, ab Z. 14). Inc. mut.: ] faciunt sic victos e contra[rio] voti impotes qui non possunt facere quod volunt … (3rb) Schlussschrift: Laus Deo. (3va) nur Rahmung und Linierung. 2 (3vb–25ra) Commentarius in Vergilii Georgica, Lib. 1–4 (Thilo III 1, 128–360). (25rb–26vb) nur Rahmung und Linierung. 3 (27ra–170va) Commentarius in Vergilii Aeneidos, unvollständig. (27ra–84vb) Lib. 1,1–Lib. 5,42 (Thilo I 7, Z. 5–594, Z. 25). Inc. mut.: ] Ausoniis disterminat arva colonis unde apparet Antenorem non ad Italiam venisse … – Expl. mut.: … non fugantur semper [ (mit Reklamante: Enim) (85ra–135vb) Lib. 5,78–Lib. 8,663 (Thilo I 601, Z. 8–II 295, Z. 27). Inc. mut.: ] gaudet purpu reosque iacit flores … – Expl. mut.: … flamines in capite [ (mit Reklamante: habebant) (136ra–170va) Lib. 10,475–Lib. 12 (Thilo II 441, Z. 7–650). Inc. mut.: ] poetis plerumque con ceditur … – Expl.: … adhuc habitare nature lege poterat sic Homerus. (170va) Schlussschrift: Explicit Servii expositio gramatici super libros Eneidos. (170vb–171vb) nur Rahmung und Linierung. 4 (172r) Federproben. 1) ‹Publius Papinius Statius›: Thebais II 620 (Ed. A. Klotz, Th. Klinnert. Lipsiae 21973). Unusne viri tot cedibus unus ibit ovans [A]rgos. 2) ‹Publius Vergilius Maro›: Georgica, Lib. 1,1, unvollständig (Ed. R. A. B. Mynors. Oxonii 1990). Quid faciat letas seg[ 3) Darunter Notiz: Bartholomei Cremonensis. Möglicherweise Hinweis auf Vergilausgabe des Bartolomeo da Cremona (Venetiis 1472 = GW M49740, ohne Servius-Kommentar) oder Besitz vermerk: In Frage kommt hierfür der vorgenannte Drucker. Eine Zuordnung zu Bartolomeo Sacchi (1421–1481), auch B. Platina und B. Cremonensis genannt, erscheint aufgrund eines Ver gleichs der Hände wenig wahrscheinlich. (172v) leer. (Ir–v) leer. P. K. 200 Cod. 174 Cod. 174 SERMONES DE TEMPORE Pap. I, 225, I Bl. 280 × 200. Tirol (?), um 1470 (u. a. 1472). B: Bl. 56rb Loch, Bl. 156r–179v tw. fleckig und Tintenfraß. Nachweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Brixen, Priesterseminarbibl., Cod. A.2; Neustift, Stiftsbibl., Cod. 204; Innsbruck, ULB Tirol, Cod. 621; Zwettl, Stiftsbibl., Cod. 205, T. II (dat. 1481). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 18.VI215(216) + (III+3)224(225) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Bl. 216–218 eingehängte Einzelbl. Nachsatzbl. mit Makulaturstreifen verstärkt (] et motibus carnalibus habuerit, s. auch bei E [VDS bzw. HDS]). Am oberen Blattrand zeitgenössische Foliierung 1–156, Forts. der Foliierung 19. Jh. durch die ULB Tirol, wiederum beginnend mit 156, daher Zahl 156 doppelt angeführt. Bl. 151 und 168 jeweils neuzeitliches gelbes Papiersignakel der ULB Tirol. S: Schriftraum und Zeilenzahl mit den Händen wechselnd. Schriftraum 210/230 × 140/155. Zwei Spalten, Bl. 1ra–179vb von Tintenlinien, ab Bl. 180ra von Blindlinien gerahmt, zu 39–46 Zeilen, Bl. 151va–154rb auf Tintenlinierung (hier zusätzliche Spalte für Angabe der Blattzahlen), ab Bl. 203ra tw. auf Blindlinierung. Bastarda von mehreren Händen: 1) Bl. 1ra–154rb, dat. 1472 (Bl. 151rb); 2) Bl. 155ra–156vb und Bl. 162va– 167vb; 3) Bl. 156Ira–162rb; 4) Bl. 168ra–224va. Am seitlichen Blattrand Korrekturen bzw. Ergänzungen, tw. von anderer zeitgenössischer Hand, häufig stark beschnitten. A: Rote Auszeichnungsstriche und Unterstreichungen, Überschriften. Kapitelzählung in tw. roten, tw. tintenfar benen arabischen Ziffern sowie rote Anmerkungen und Hervorhebungen am Blattrand. Rote meist drei-, gelegentlich vierzeilige Lombarden, tw. mit Punktverdickungen oder kurzen Fadenausläufern, vereinzelt einfachen Aussparungen. Buchstabenschäfte am Rand des Schriftraums bisweilen gelängt, vereinzelt mit einfachem Dekor, Bl. 201va Tierdarstellung in Federzeichnung, Bl. 204ra cadellenartige, rubrizierte Ober länge mit Profilgesicht. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. etwas abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck HOMILIA IN DOMINIC. in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), darunter Makula turstreifen. Spiegel des HD leer, darunter Exlibris sowie Makulaturstreifen. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh., dasselbe Exlibris auch verkehrt unter dem Spiegel des HD erkennbar. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Si gnaturen der ULB Tirol II 2 H 22 (Tinte, Bleistift) und Nr. 174/I (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 174/I. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 143, 208. – Stefani 105. – Peintner, Stiftsbibliothek 121. 1 1.1 1.2 (Ir–v) leer. (1ra–151rb) Sermones de epistolis de tempore „Sensatus“. (1ra–4rb) Konkordanz I zw. Festtag, Thema und Auswahl der Sermones für Predigten über das Evangelium. Tit.: Tabulacio quomodo ad predicandum sit in hoc opere de ewangeliis. – Inc.: Nota. Si alicui plus placuerit predicare de ewangeliis quam de epistolis … – Expl.: … sic de aliis procedas de quibus autem placurit (!) predicare etc. (4v–5r) konkordanz II (gekürzt). Tit.: Incipit recapitulacio distinccionum huius libri. – Inc.: Distinccio 1a Christus est nobis prope quattuor modis … – Expl.: … 60 Oracio nostra ut exaudiatur a Deo debet esse quadruplex dis posita. (5v) leer. Cod. 174 201 1.3 (6ra–151rb) Sermones de tempore „Sensatus“ (Schneyer 5, 358–362, Nr. 1–60). (6ra) Prolog (Nr. 1). – (6ra) Dnca 1. adv. (Nr. 2). – (8ra) Dnca 2. adv. (Nr. 3). – (9va) Dnca 3. adv. (Nr. 4). – (11rb) Dnca 4. adv. (Nr. 5). – (14vb) In vigilia nativitatis dni (Nr. 6). – (16rb) In nati vitate dni (Nr. 7). – (18ra) Dnca infra octav. nativitatis dni (Nr. 8). – (20ra) Dnca 1. post epiph. dni (Nr. 9). – (21va) Dnca 2. post epiph. dni (Nr. 10). – (24ra) Dnca 3. post epiph. dni (Nr. 11). – (26rb) Dnca 4. post epiph. dni (Nr. 12). – (29ra) Dnca 5. post epiph. dni (Nr. 13). – (32vb) Dnca in Septuages. (Nr. 14). – (36rb) Dnca in Sexages. (Nr. 15). – (38va) Dnca in Quinquages. (Nr. 16). – (41rb) In capite ieiunii (Nr. 17). – (43ra) Dnca 1. in Quadrages. (Nr. 18). – (44vb) Dnca 2. in Quadrages. (Nr. 19). – (46va) Dnca 3. in Quadrages. (Nr. 20). – (49ra) Dnca 4. in Quadrages. (Nr. 21). – (51rb) Dnca 1. in passione dni (Nr. 22). – (53ra) Dnca in palmis (Nr. 23). – (55vb) Fer. 5. in coena dni (Nr. 24). – (58rb) Fer. 6. in parasceve (Nr. 25). Tit.: In passione domini. – (60va) Dnca resurrectionis dni (Nr. 26). – (63vb) Dnca 1. post pascha (Nr. 27). – (64vb) Dnca 2. post pascha (Nr. 28). – (67vb) Dnca 3. post pascha (Nr. 29). Tit.: Dominica 2a post dominicam pasche (!). – (70va) Dnca 4. post pascha (Nr. 30). Tit.: Dominica 3a post pasce. – (72vb) Dnca 5. post pascha (Nr. 31). Tit.: Dominica 4a post pascha. – (75ra) In ascensione dni (Nr. 32). – (77va) Dnca infra oct. ascensionis dni (Nr. 33). – (80rb) Dnca pentecostes et de spiritu sancto (Nr. 34). – (82vb) In festo ss. trinitatis (Nr. 35). – (85va) Tit.: De corpore Christi (Nr. 36). – (87va) Dnca 2. post pent. (Nr. 37). Tit.: Dominica prima post Penthecostes. (90vb) Tit.: De morte corporali et spirituali per totum. Dominica secunda. – Inc.: Translati sumus de morte ad vitam. Ia Ioh. 4a (3,14). Nos enim videmus ad sensum quod quando aliqui evaserunt mortem qui fuerunt in bello fortissimo ubi multi mortui fuerunt ad suos dilectos veniunt et dicunt … – Expl.: … quasi post conbustionem ab igne liberari. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 151, Bl. 81vb. (93va) Dnca 3. post pent. (Nr. 38). – (97rb) Dnca 4. post pent. (Nr. 39). – (99ra) Dnca 5. post pent. (Nr. 40). – (100rb) Dnca 6. post pent. (Nr. 41). – (102rb) Dnca 7. post pent. (Nr. 42). Tit.: Dominica 7a post (gestrichen: Penthe) Trinitatis. – (104rb) Dnca 8. post pent. (Nr. 43). – (106va) Dnca 9. post pent. (Nr. 44). – (108vb) Dnca 10. post pent. (Nr. 45). – (111rb) Dnca 11. post pent. (Nr. 46). – (114ra) Dnca 12. post pent. (Nr. 47). – (116va) Dnca 13. post pent. (Nr. 48). – (119ra) Dnca 14. post pent. (Nr. 49). – (122va) Dnca 15. post pent. (Nr. 50). – (125ra) Dnca 16. post pent. (Nr. 51). Expl.: … qualis est sua dileccio. – (129ra) Dnca 17. post pent. (Nr. 52). – (131rb) Dnca 18. post pent. (Nr. 53). – (133rb) Dnca 19. post pent. (Nr. 55). – (135vb) Dnca 20. post pent. (Nr. 54). – (138va) Dnca 21. post pent. (Nr. 56). – (140va) Dnca 22. post pent. (Nr. 57). – (142va) Dnca 23. post pent. (Nr. 58). – (145vb) Dnca 24. post pent. (Nr. 59). – (148vb) Dnca 25. post pent. (Nr. 60). (151rb) Datierung: 1472. 1.4 (151va–154rb) Thematisches Register in alphabetischer Reihenfolge mit Blattangabe. Inc.: Activa et contemplativa vita 30. Angeli gaudent de conversione peccatoris 31… – Expl.: … Virgines habebunt 4or singularia in patria 127. Virtutes comparantur luci triplici racione 71. (154v) leer. 2 (155ra–167vb) Sermones de tempore, größtenteils anonym. (155ra) De annuntiatione BMV. Inc.: Ave gracia plena dominus tecum. Luc. 1 (28). Ista dies precipue maior est omnibus festivitatibus … – Expl.: … induit pro redempcione humani generis in cruce obligavit. Rogemus dominum ut nobis ferat eternum premium. (156ra) Tit.: Sermo de corpore Christi. In cena domini (Schneyer 8, 419, Nr. 81). (156Ira) ‹Ambrosius Spiera›: Sermo Fer. 6 in parasceve (Schneyer II Nr. 97) bzw. ‹Albertus Engelschalk› (Schneyer II Nr. 323). Inc.: Egressus Ihesus trans torrentem Cedron ubi erat ortus in quem introivit et ipse cum discipuli (!) suis (Io 18,1). Sicut enim tunc vos scitis, quod mos predicatorum est in principio sermonum predicatorum ad beatam virginem recurrere … – Expl.: … in commendacionem sue caritatis in augmentum mee devocionis et in meritum vite eterne. 202 Cod. 174 Inc. wie Graz, Franziskanerkloster, Cod. A 67/29, Bl. 100va. (156Ira) Inc.: Egressus Ihesus trans torrentem Cedron (Io 18,1). Quando enim occurrat festum alicuius sancti tunc consuevimus eius legendam et passionem ut Laurencii Petri etc. Sic enim hodie est flendum … – Expl.: … eligendo flores de IIIIor ewangelistis. Inc. vgl. Graz, Franziskanerkloster, Cod. A 67/29, Bl. 100va. (156Ira) Historia passionis dni secundum quattuor evangelistas. Inc.: Historia passionis Christi fuit per istum modum sicut tunc dicit beatus Iohannes ewangelista qui tunc passionem filii Dei scripsit … – Expl.: … vero Christus sepultus fuisset revertebatur virgo Maria amarissime plo rans secundum Augustinum procede. (162va) Tit.: Exhortacio in die palmarum. – Inc. (ohne Bibelthema): Hodie sancta mater ecclesia duas processiones recolit … – Expl.: … vicit […] claustrum […]. (163ra) Inc.: Sicut ovis ad occisionem ductus est. Is. 53 (7). Hodie karissimi dies celebratur insignis et festiva in tanto maioris devocionis affectu a singulis Christi fidelibus est celebranda … – Expl.: … et post hanc vitam celestem gloriam intrare et possidere meremur quod nobis concedere dignetur Ihesus Christus dominus noster qui eam nobis suo sanguine comparavit qui est bene dictus in sanguine suo Amen. Inc. wie Augsburg, UB, Cod. II. 1. fol. 48, Bl. 81rb. (166va) Inc.: Qui manducat hunc panem vivet in eternum. Ioh. 6 (58) et in ewangelio hodierne leccionis. Magnam hodie et precipuam karissimi omnique devocione recollendam agimus so lempnitatem ad cuius celebracionem venerandam venerabilis Hugo de Sancto Victore in quodam sermone ipsius sollempnitatis nos devote invitat … – Expl.: … ex hoc salutem consequere hic per graciam et in futuro per gloriam Amen. 3 (168ra–224vb) Liber sermonum „Fons vitae“ (Schneyer 7, 611–617, Nr. 1–65). Festangabe bei Schneyer tw. abweichend; bei Abweichungen in der Hs. jene bei Schneyer in Klammer angeführt. (168ra) Tit.: Dominica prima in adventu domini. – Inc.: Hora est iam nos de sompno surgere. Apostolus ad Romanos (13,11). Egregius predicator et apostolus invitans nos ad opera virtutum. In istis verbis tria facit. Primo allegat temporis oportunitatem … – Expl.: … et ipsum amicum tuum faciunt. Rogemus. (169rb) Tit.: Sermo 2us de eodem (Schneyer 7, Nr. 1). – (170rb) Tit.: Dominica secunda (Nr. 2). – (171va) Tit.: Dominica tercia (Nr. 3). – (172rb) Tit.: Dominica quarta (gestrichen: quinta) (Dnca 3. adv., Nr. 4). – (173rb) Tit.: In nativitate domini (Nr. 5). – (174rb) Tit.: Sermo de eodem (Dnca infra octav. nativitatis dni, Nr. 6). – (175rb) Tit.: Dominica prima post epiphaniam (Nr. 7). – (176vb) Tit.: Dominica secunda post epiphaniam (Nr. 8). – (177vb) Tit.: Dominica tercia (Nr. 9). – (179ra) Tit.: Dominica quarta (Nr. 10). – (180ra) Dnca 5. post epiph. dni (Nr. 11). – (180vb) Dnca in Septuages. (Nr. 12). – (181va) Dnca in Sexages. (Nr. 13). – (182ra) Dnca in Quinquages. (Nr. 14). – (182va) Dnca 1. in Quadrages. (Nr. 15). – (183rb) Dnca 2. in Quadrages. (Nr. 16). – (184rb) Dnca 3. in Quadrages. (Nr. 17). – (185ra) Dnca 4. in Quadrages. (Nr. 18). – (186ra) Dnca 1. in passione dni (Nr. 19). – (186vb) Dnca in palmis (Nr. 20). – (187rb) Fer. 5 in coena dni (Nr. 21). Inc.: Acccippe (!) et manducate ex hoc omnes Actis Ioh. (recte: 1 Cor 11,24). Nota quando tritulator est debilis et habet grave flagellum in manu … Inc. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 277v. – (188va) Fer. 6. in parasceve (Nr. 22). – (189rb) Nr. 23. – (189vb) In vigilia paschatis (Nr. 24). – (191ra) Dnca resurrectionis dni (Nr. 25). Expl.: … dixit de templo corporis sui etc. Rogemus. Expl. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 281v. – (191vb) Nr. 26. Expl.: … tercia autem conversionem in bono perseverancium. Rogemus ergo dominum. Expl. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 281v. – (192va) Tit.: In octava resurreccionis (Dnca 1. post pascha, Nr. 27). – (193va) Tit.: Dominica prima post octavas (Dnca 2. post pascha, Nr. 28). – (194va) Tit.: Dominica secunda (Dnca 3. post pascha, Nr. 29). – (195ra) Tit.: Dominica tercia (Dnca 4. post pascha, Nr. 30). – (196ra) Tit.: Dominica quarta (Dnca 5. post pascha, Nr. 31). Inc. bei Schneyer unsichere Lesung, in der Hs.: … quia promptus sum … – Expl. bei Cod. 177 203 Schneyer unsichere Lesung, in der Hs.: … vel tribulacio a dyabolo detestinata. – (196vb) Tit.: Dominica quinta (In Litaniis maioribus et minoribus, Nr. 32). – (197va) Tit.: In die sancto ascensionis domini (Nr. 33). – (198rb) Tit.: Dominica post ascensionem domini (Dnca 6. post pascha, Nr. 34). – (198vb) Tit.: De eodem (Nr. 35). – (199va) Tit.: De sancto Penthecostes (Nr. 36). – (200ra) Tit.: De eodem (Nr. 37). – (200vb) Ohne Festangabe bei Schneyer (Nr. 38). – (201va) Tit.: De sancta trinitate (Nr. 39). – (202va) Dnca 1. post pent. (Nr. 40). – (204ra) Dnca 2. post pent. (Nr. 41). Expl.: … et fons perpetue consolacionis. Rogemus ergo dominum. Expl. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 296r. – (204va) Dnca 3. post pent. (Nr. 42). (205va) Tit.: Dominica quarta. – Inc.: Estote misericordes sicut et pater vester misericors est. Luc. VIo (36). Nota in precedenti ewangelio dominicali dicitur dicitur (!) quod publicani et pec catores … – Expl.: … pauperibus distribuendo centuplum accipiet in vitam eternam. Rogemus. Parallelüberlieferung: Kremsmünster, Stiftsbibl., CC 23, Bl. 297v. (206va) Tit.: Dominica quinta (Nr. 43). – (207va) Tit.: Dominica sexta (Dnca 5. post pent., Nr. 44). Expl.: … velox ad iram quia occisio aliquando sequitur. Rogemus. Expl. wie Krems münster, Stiftsbibl., CC 23, Bl. 300r. – (208rb) Tit.: Dominica VII (Dnca 6. post pent., Nr. 45). – (209ra) Tit.: Dominica octava (Nr. 46). – (209va) Tit.: Dominica nona (Dnca 7. [!] post pent., Nr. 47). Expl.: … Sic est deliciosus. Rogemus etc. Expl. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 302v. – (210va) Tit.: Dominica decima (Dnca 8. post pent., Nr. 48). – (211rb) Tit.: Undecima dominica (Dnca 9. post pent., Nr. 49). – (212va) Tit.: Dominica XII (Dnca 10. post pent., Nr. 50). – (213rb) Tit.: Dominica XIII (Dnca 10. post pent., Nr. 51). – (213vb) Tit.: Dominica XIIII (Dnca 11. post pent., Nr. 52). – (215ra) Tit.: Dominica XV (Dnca 12. post pent., Nr. 53). – (215va) Tit.: Dominica quartadecima (Dnca 13. post pent., Nr. 54). – (216rb) Tit.: Dominica quintadecima (Dnca 14. post pent., Nr. 55). – (217ra) Tit.: Dominica sedecima (Dnca 15. post pent., Nr. 56). – (217vb) Tit.: Dominica XVII (Dnca 16. post pent., Nr. 57). – (218va) Tit.: Dominica XVIII (Dnca 17. post pent., Nr. 58). Expl.: … manus eius contra omnes. Genesis. Rogemus ergo etc. Expl. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 313v. – (219ra) Tit.: Dominica XVIIII (Dnca 18. post pent., Nr. 59). – (219va) Tit.: Vicesima dominica (Nr. 60). Expl.: … sustinere debuit. Mt. ultimo Accipiens etc. Rogemus. Expl. vgl. Kremsmünster, Stiftsbibl., CC 23, Bl. 314r. – (220ra) Tit.: Vicesima prima (Dnca 20. post pent., Nr. 61). – (221ra) Tit.: Vicesima secunda (Dnca 21. post pent., Nr. 62). – (222ra) Tit.: Vicesima tercia (Dnca 22. post pent., Nr. 63). – Inc.: Ostendite michi numisma census. Mt. XXIIo (19). Nota predicatores dicuntur pastores quibus dicetur (!) Iohannes XXIo Afferte michi de piscibus … Inc. wie Kremsmünster, Stiftsbibl., CC 23, Bl. 316v. – (223ra) Tit.: Vicesima quarta (Dnca 23. post pent., Nr. 64). – (223va) Tit.: Dominica vicesima quinta (Dnca 24. post pent., Nr. 65). (224r–v) leer. (Ir–v) leer. A. P. / U. S. Cod. 177 FLORES DE SANCTIS ET DE TEMPORE Pap. I, 153, I Bl. 280 × 210. Tirol (Neustift?), um 1430/33. B: Pap. tw. fleckig, bes. Bl. 41–47, Bl. 12 am seitlichen Blattrand eingerissen. Bl. 33 am seitlichen Blattrand beschnitten. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 481 (dat. 1431), Cod. 437, T. II (dat. 1432), Cod. 666, T. III (dat. 1433). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + V10 + 212 + 2.V32 + 133 + 4.VI81 + (VI–1)92 + 5.VI152 + 1153 + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 83 herausgeschnitten (ohne Textverlust). Lage 4 und 9 mit einem 204 Cod. 177 Papierfalz umschlagen. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: Bl. 152 und 153 fälschlich als 252 und 253 foliiert, später korrigiert. S: Schriftraum 180/200 × 130. Zwei Spalten, von Tintenlinien gerahmt, zu 33–38 Zeilen. Zirkelstiche. Bastarda von drei Händen: 1) Bl. 1ra–80va, Z. 30; 2) Bl. 80va, Z. 30–Bl. 106vb, Z. 9 und Bl. 153ra–va; 3) Bl. 106vb, Z. 10–Bl. 152vb. Randbemerkungen von späterer Hand. A: Rote Auszeichnungsstriche, Unterstreichungen und Überschriften. Zeigehände. Oberlängen in der ersten Zeile bisweilen gedehnt. Drei- und vierzeilige (Bl. 1ra und 140ra achtzeilige) rote Lombarden, tw. mit ungelenken kopfstempelartigen Aussparungen, tw. mit Punktverdickungen, tw. mit minimal verzierten Ausläufern. Ver einzelt auch fünfzeilige rote Lombarden mit einfachem Knospenfleuronnée als Besatz. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. FLORES 1460 in Goldrahmung, im dritten Si gnaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 1r Besitzvermerk 18. Jh. Collegii Neocell. Anno 1460. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD Signaturen der ULB Tirol II (Tinte) 2 K 25 (Bleistift) und Nr. 177 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 177. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Ger main an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 158, 208. (Ir–v) leer. (1ra–143rb) Flores (Graeculus Piper) secundum Conradum de Waldhausen bzw. Graeculus (Schne yer 1, 805–815, Nr. 149–326 bzw. Schneyer 2, 206–219, Nr. 1–174). Tit. am oberen Blattrand: Flores de sanctis et de tempore simul. Autorenschaft unsicher. Im Folgenden Sermones nach Schneyer für Conradus de Waldhausen (Schneyer 1) und Graeculus (Schneyer 2) zitiert; drei Sermones der Hs. fehlen bei Graeculus, eine bei Conradus de Waldhausen. Wenn nicht anders angegeben, wird zunächst jeweils die entsprechende Nummer in Schneyer 1, dann jene in Schneyer 2 angeführt. (1ra) Tit.: Dominica in adventu domini prima etc. (Schneyer 1, 805, Nr. 149 für Conradus de Wald hausen bzw. Schneyer 2, 206, Nr. 1 für Graeculus). (1vb) Schneyer 1, Nr. 150 bzw. Schneyer 2, Nr. 2. (2vb) Nr. 151 bzw. Nr. 3. (4ra) Nr. 152. (5ra) Nr. 153 bzw. Expl. eher Nr. 4. (6ra) Nr. 154 bzw. Nr. 5. (7ra) Dnca 2. adv. (Nr. 155). (7va) Tit.: Sermo tercius (Nr. 156 bzw. Nr. 6). (8va) Tit.: Dominica tercia in adventu (Expl. eher Nr. 157 bzw. Nr. 7). (9rb) Nr. 158 bzw. eher Nr. 8. (10va) Nr. 160. (11va) Dnca 4. adv. (Nr. 161 bzw. Nr. 10). (12rb) Nr. 162 bzw. Nr. 11. (13rb) Nr. 163 bzw. Nr. 12. (14ra) Nr. 164 bzw. Expl. eher Nr. 14. (15ra) Tit.: In nativitate domini (Nr. 165). (16ra) Nr. 166 bzw. Nr. 15. (17rb) Nr. 167 bzw. Nr. 16 mit leicht abweichendem Inc. (18ra) Nr. 168 (nicht in Schneyer 2). (19rb) Nr. 169 (nicht in Schneyer 2). (20va) Nr. 170 bzw. Nr. 17. (21va) Dnca infra octav. nativitatis dni (Nr. 171 bzw. Nr. 18). (22va) Nr. 173 bzw. Nr. 19 mit leicht abweichendem Inc. Expl. (= Expl. Nr. 175 bzw. Nr. 22): … Unde Augustinus Quicumque se ad hoc nomen devote inclinaverit decem dies indulgencie consequitur. Invoca ergo hoc nomen Ihesus ut con sequaris remissionem omnium peccatorum. (23rb) In circumcisione dni. (Inc. wie Nr. 175 und 176 bzw. Nr. 21). Expl. abweichend (ähnlich Nr. 176 bzw. Nr. 21): … Invoca nomen eius quod hodie sibi impositum est in quo quidem nomine solum salvari poteris ut dicit apostolus. (24vb) Tit.: In epiphania domini sermo (Schneyer 1, Nr. 177). (25vb) Nr. 178 bzw. Nr. 23. (26vb) Nr. 179 (nicht in Schneyer 2). Expl.: … nam communis est omnibus divitibus et pauperibus dare reg num celorum. Rogemus igitur puerum Ihesum Alleluia. (28ra) Nr. 180. Bibelzitat leicht abweichend (bei Schneyer Mt 2,11 beginnend mit „Apertis thesauris suis“, in der Hs. Mt 2,11 erst ab): Obtulerunt Cod. 177 205 domino aurum … (28vb) Nr. 181 bzw. Nr. 25. (30ra) Dnca 1. post epiph. dni (Nr. 182 bzw. Nr. 26). (31ra) Nr. 183 bzw. Nr. 27. Expl.: … obsequendo et amando invenit et puerum Ihesum placabilem sibi. Rogemus igitur dominum. (31vb) Schneyer 2, Nr. 28. (32va) Nr. 185 bzw. Nr. 30. (33vb) Dnca 2. post epiph. dni (Nr. 186 bzw. Nr. 31). Expl. erweitert: … sic per mulierem diligentem ornatur domus (= Ende Schneyer). Ergo diligere debemus uxores etc. (34rb) Nr. 187 bzw. Nr. 32. (35va) Dnca 3. post epiph. dni (Nr. 188 bzw. Nr. 33). (36vb) Nr. 189 bzw. Nr. 34. (38rb) Nr. 190 bzw. Nr. 35. (39rb) Dnca 4. post epiph. dni (Nr. 191 bzw. Expl. eher Nr. 36). (40vb) Expl. eher Nr. 192 bzw. Nr. 37. (42ra) Nr. 193 bzw. Expl. eher Nr. 38. (43rb) Dnca in Septuages. (Nr. 196 bzw. Nr. 41). Am Rand Bemerkung Valde popularis et utilis ad docendum. (44ra) Nr. 197 bzw. Nr. 42. (44vb) Dnca 5. post epiph. dni (Nr. 194 bzw. Nr. 39). Inc. jeweils leicht abweichend. Inc.: Domine, nonne bonum semen seminasti etc. Matth. (13,27). Plures sunt agri in quibus seminat … (46ra) Nr. 195 bzw. Nr. 40. (47rb) Dnca in Sexages. (Nr. 198 bzw. Nr. 43). (48rb) Bei Schneyer 1 nur Bibelzitat wie Nr. 199 bzw. Inc. Nr. 44. Expl.: … Confitentur se nosce Deum factis autem negant. (49rb) Dnca in Quinquages. (Nr. 200 bzw. Nr. 45). Expl. (jeweils leicht abweichend): … sic et quilibet debet videre intra se qui debeat mundare. (50ra) Nr. 201 bzw. Inc. und Expl. eher Nr. 46. (51rb) Nr. 202 bzw. Expl. eher Nr. 47. (52ra) Tit.: Feria quarta cinerum (Nr. 203 bzw. Expl. eher Nr. 49). (53ra) Nr. 204 bzw. Nr. 50. (54ra) Nr. 205 bzw. Nr. 51. (55ra) Nr. 206 bzw. Expl. eher Nr. 52. (56va) Dnca 1. in Quadrages. (Nr. 207, Expl. eher Nr. 53). Expl.: … ad infernum detraheris in profun dum laci. (57vb) (Nr. 208 bzw. Nr. 54). (59rb) Dnca 2. in Quadrages. (Nr. 209 bzw. Nr. 55). (60rb) Inc. von Nr. 210 leicht abweichend bzw. eher Nr. 56. (61rb) Nr. 211 bzw. Nr. 57. (62rb) Dnca 3. in Quadrages. (Expl. eher Nr. 212 bzw. Nr. 58). (63rb) Expl. eher Nr. 213 bzw. Nr. 59. (64ra) Expl. eher Nr. 214 bzw. Nr. 60. (64vb) Dnca 4. in Quadrages. (Nr. 215 bzw. Nr. 61). (66ra) Nr. 216 bzw. Nr. 62. (66vb) Nr. 217 bzw. Nr. 63. (67vb) Mit dt. Zusatz: … quia numquam liberabor. Und reimet sich das merlin nit wol so iber doch der sinn gern guot. Rogemus etc. (67vb) Dnca 1. in passione dni (Nr. 218 bzw. Nr. 64). (68vb) Nr. 219 bzw. Nr. 65. (69vb) Nr. 220 bzw. Expl. eher Nr. 66. (70rb) Dnca in palmis (Nr. 221 bzw. Nr. 67). (71rb) Nr. 222 bzw. Nr. 68. (72rb) Nr. 223 bzw. Nr. 69. (73rb) Tit.: In cena domini sermo (Nr. 225 bzw. Nr. 71). Expl. erweitert: … In pedum ablucione innuitur (= Ende Schneyer) humilitas ubi dicitur quod humilitas est summa virtus. (73vb) Nr. 227. (75rb) Tit.: Idem. Nur Bibelzitat wie Nr. 226, Inc. wie Nr. 72. Inc.: Ad mensam magnam sedisti (Sir 31,12). Scito quod talia oportet te preparare qualia sunt in hostia. Quanto maior est festivitas tanto se melius homines preparare debent … – Expl.: … Tales non debent accipere corpus Christi. Rogemus. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 758, Bl. 189v. (76va) Tit.: In parascephenn (!) (Nr. 228 bzw. Nr. 73). (78rb) Nr. 230 bzw. Nr. 75. (78vb) Tit.: In die sancto pasche (Nr. 231 bzw. Nr. 76). (80rb) Nr. 232 bzw. Expl. eher Nr. 77. (81ra) Nr. 233 bzw. Nr. 78. (82ra) Nr. 234 bzw. Nr. 79. (82rb) Dnca resurrectionis dni, fer. 2 (Nr. 235 bzw. Nr. 80). (83vb) Nr. 236 bzw. Nr. 81. (84rb) Expl. eher Nr. 237 bzw. Nr. 82. (85rb) Dnca resurrectionis dni, fer. 3 (Nr. 240 bzw. Nr. 83). (86ra) Dnca 1. post pascha (Nr. 241 bzw. Nr. 84). (86va) Nr. 243 bzw. Inc. eher Nr. 85. (87rb) Nur Bibelzitat und Expl. wie Nr. 242, Inc. übereinstimmend mit Nr. 86. (88ra) Dnca 2. post pascha (Nr. 244 bzw. Nr. 87). (88vb) Nr. 245 bzw. Nr. 88. (89va) Nr. 246 bzw. Nr. 89. (90ra) Dnca 3. post pascha (Nr. 247 bzw. Nr. 90). (91ra) Nr. 248 bzw. Nr. 91. (91vb) Nr. 249 bzw. Nr. 92. 206 Cod. 177 (92va) Dnca 4. post pascha (Nr. 251 bzw. Inc. eher Nr. 94). (93va) Nr. 252 bzw. Nr. 95. (94ra) Nr. 253 bzw. Nr. 96. (95ra) Dnca 5. post pascha (Nr. 254 bzw. Nr. 97). (96ra) Nr. 255 bzw. Nr. 98. (96va) Expl. eher Nr. 256 bzw. Nr. 99. (97ra) Nr. 257 bzw. Nr. 100. (97vb) Nr. 259 bzw. Nr. 101. (98va) In ascensione dni (Nr. 260 bzw. Nr. 102). (99va) Nr. 261 bzw. Nr. 103. (100va) Nr. 262 bzw. Nr. 104. (101rb) Dnca 6. post pascha (Nr. 263 bzw. Nr. 105). (102rb) Nr. 264 bzw. Nr. 106. (103rb) Nr. 265 bzw. Nr. 107. (104rb) Dnca pentecostes et de spiritu sancto (Nr. 266 bzw. Nr. 108). (105ra) Nr. 267 bzw. Nr. 109. (105va) Nr. 268 bzw. Nr. 110. (106vb) In festo ss. trinitatis (Nr. 269 bzw. Expl. eher Nr. 111). (107rb) Nr. 270 bzw. Expl. eher Nr. 112. (108rb) Dnca 1. post pent. (Inc. wie Nr. 271, Expl. Nr. 272 bzw. Inc. wie Nr. 113, Expl. Nr. 114). (109va) Nr. 273 bzw. Nr. 115. (110va) Dnca 2. post pent. (Nr. 276 bzw. Nr. 116). (111rb) Nr. 274 bzw. Nr. 117. (112rb) Nr. 275 bzw. Nr. 118. (113ra) Dnca 3. post pent. (Nr. 277 bzw. Nr. 119). (114ra) Nr. 278 bzw. Nr. 120. (114vb) Expl. eher Nr. 279 bzw. Nr. 121. (115vb) Dnca 4. post pent. (Nr. 280 bzw. Nr. 122). (116va) Nr. 281 bzw. Nr. 123. (117va) Inc. mit Glosse wie Nr. 282 bzw. Nr. 124. (118rb) Dnca 5. post pent. (Nr. 283 bzw. Nr. 125). (119rb) Nr. 284 bzw. Nr. 126. (120ra) Nr. 285 bzw. Nr. 127. (120vb) Dnca 6. post pent. (Expl. eher Nr. 286 bzw. Nr. 128). (121va) Nr. 287 bzw. Nr. 129. (122rb) Nr. 288 bzw. Nr. 130. (123rb) Dnca 7. post pent. (Expl. eher Nr. 289 bzw. vgl. Nr. 131). (124ra) Nr. 291 bzw. Nr. 132. (124va) Nr. 290 bzw. Nr. 133. (125va) Dnca 8. post pent. (Expl. eher Nr. 292 bzw. Nr. 134). (126rb) Expl. Nr. 293 bzw. Nr. 135. (127ra) Nr. 294 bzw. Nr. 136. Expl. leicht gekürzt: … ceteris peccantibus tantum dat unam. (127va) Dnca 9. post pent. (Nr. 295 bzw. Nr. 137). (128rb) Nr. 296 bzw. Nr. 138. (129ra) Nr. 297 bzw. Nr. 139. (129vb) Dnca 10. post pent. (Nr. 298 bzw. Nr. 140). (130va) Nr. 299 bzw. Nr. 141. (131rb) Nr. 300 bzw. Nr. 142. (132ra) Dnca 11. post pent. (Nr. 301 bzw. Nr. 143). (133ra) Nr. 302 bzw. Nr. 144. (133vb) Nr. 303 bzw. Nr. 145. (134rb) Dnca 12. post pent. (Nr. 304 bzw. Nr. 146). Inc. leicht abweichend: Misit digitos suos … Mt. 4o (recte: Mc 7,33). Digitos in aures surde (!) et tandem rogaverunt tam ut imponeret sibi manum … (134vb) Nr. 305 bzw. Nr. 147. (135va) Dnca 13. post pent. (Nr. 306 bzw. Nr. 148). (136rb) Nr. 307 bzw. Nr. 149. (137rb) Nr. 308 bzw. Nr. 150. (137vb) Dnca 14. post pent. (Nr. 309 bzw. Nr. 151). (138rb) Nr. 310 bzw. Nr. 152. (138vb) Dnca 15. post pent. (Nr. 311 bzw. Nr. 153). (139va) Expl. eher Nr. 312 bzw. Nr. 154. (140ra) Nr. 313 bzw. Nr. 155. (140vb) Nr. 314 bzw. Nr. 156. (141rb) Dnca 16. post pent. (Nr. 315 bzw. Nr. 157). (142rb) Nr. 316 bzw. Nr. 158. (143ra) Nr. 317 bzw. Nr. 159. (143rb) Dnca 17. post pent. (nur Bibelzitat und Expl. vgl. Nr. 318 bzw. Inc. wie Nr. 160). (144rb) Nr. 319 bzw. Nr. 161. (144vb) Nr. 320 bzw. Nr. 162. (145va) Dnca 18. post pent. (Nr. 321 bzw. Expl. eher Nr. 163). (146rb) Dnca 18. post pent. (Expl. eher Nr. 322 bzw. Nr. 164). (147rb) Dnca 19. post pent. (Nr. 166). (148rb) Nr. 167. (148vb) Nr. 168) (149va) Dnca 20. post pent. (Nr. 170). Expl.: … Math. Venite benedicti patres etc. (150ra) Nr. 324 bzw. Nr. 169. Cod. 181 207 (151ra) Dnca 21. post pent. (Nr. 171). (151vb) Nr. 325 bzw. Nr. 172. (152va) Nr. 173. (153rb) Dnca 22. post pent. (Nr. 326 bzw. Nr. 174). Expl.: … et submergit scriptura humilitatem eius dicens. (153vb) leer. (Ir–v) leer. U. S. Cod. 181 ALANUS AB INSULIS Pap. I, 60, I Bl. 285 × 210. Wiener Raum, um 1410. B: Pap. bisweilen stark fleckig, Wasserschaden, Risse und Fehlstellen, nachträglich ergänzt, Lage 1 mit Pap. verstärkt. Zw. Lage 1 und 2 sowie in den Lagenmitten Pergamentfälze. Bilder und Nachweise der Wasser zeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 5.VI60 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Lagenreklamanten. Am Beginn von Lage 2 Kustode in arabischer Ziffer erhalten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 200/210 × 140/145, von tw. kaum sichtbaren Bleistiftlinien begrenzt. Zeilenzahl wechselnd. Bas tarda. Incipit in Auszeichnungsschrift (Textura). A: Rote Auszeichnungsstriche, Unterstreichungen, Rahmungen der Über- und Schlussschriften. Zeigehände. In der ersten Zeile bisweilen cadellenartige Verlängerung der Schäfte einzelner Buchstaben und einfache Verzierungen (in Federzeichnung Bl. 8rb Gesicht, Bl. 21ra Profilkopf). Zweizeilige rote Lombarden, tw. mit Punktverdickungen. Bl. 1ra einfache rote Initiale mit kopfstempelförmiger Aussparung und einer Ähre aus gestielten Perlen im Binnenfeld. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. ALAN. DE ARTE PRAED. in Goldrahmung, im dritten Signatur schild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistift notiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Innsbruck, ULB Tirol, Cod. 157, Bl. Ir gehört Cod. 181 zu einer Gruppe von urspr. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wur den. Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzelhandschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signa turen (in der urspr. Reihenfolge): Cod. 157: Aristoteles: Ethica Nicomachea und Oeconomica. heute nicht mehr nachweisbar: Aristoteles: Politica. Cod. 159: Albertus de Saxonia: Kommentar zu Cod. 157. Cod. 168: Aristoteles: Physica, Lib. 1–5. Cod. 163: Aristoteles: Physica, Lib. 6–8 und De anima. Cod. 181: Alanus ab Insulis: Summa de arte praedicatoria. Cod. 164: Anshelmus Havelburgensis. Anticimenon. Von diesen sieben Hs. ist eine nicht mehr erhalten, drei befinden sich heute an der ULB Tirol in Innsbruck (Cod. 157, 159 und 164), drei werden in der Stiftsbibl. Neustift (Cod. 163, 168 und 181) aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II`4 D 9 (Tinte, radiert), II 2 H 29 (Bleistift) und Nr. 181 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedens vertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Neuhauser, Neustift 91–95. – Katalog ULB Tirol II 167, 208. 208 Cod. 182 (Ir–v) leer. (1r–60v) Alanus ‹ab Insulis›: Summa de arte praedicatoria (PL 210, 111A–195D). Zw. cap. 30 und 31 eingeschoben 31m De penitencia (Bl. 41rb–42ra), deshalb im Folgenden Zählung von PL abweichend. Cap. 48 fehlt. (1r) Tit.: Incipit Alanus de arte predicandi. – Inc.: Vidit Iacobus scalam a terra usque ad celum attingentem per quam ascendebant … – Expl.: … et alibi. Et regnis eius non erit finis. (41rb) Einschub. Tit.: De penitencia. – Inc.: Dominus dicit in ewangelio Penitenciam agite ap propinquabit enim regnum celorum (Mt 3,2). Petrus apostolus dicit Penitenciam agite et bapti zetur … – Expl.: … docebo iniquos vias. (59r) Schlussschrift: Explicit summa magistri Alani de modo predicandi. (59v–60v) Register. (Ir–v) leer. C. S. Cod. 182 PS.-EUSEBIUS Pap. I, 16, I Bl. 285 × 205. Süddeutscher Raum, 1400. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + VIII16 + (I–1)I(Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am Lagenanfang Kustode 21, am rechten oberen Blattrand Foliierung 15. Jh. 216–231, Hinweise auf ehem. Sammelhandschrift (s. bei G). Am oberen Blattrand mittig Foliierung 19. Jh. durch die ULB Tirol 1–16 (hier berücksichtigt). S: Schriftraum 220/225 × 170/175. Zwei Spalten, von Tintenlinien gerahmt, zu 35–40 Zeilen. Bastarda, dat. 1. Oktober 1400 (Bl. 16rb). A: Rote Auszeichnungsstriche, Durchstreichungen und Überschriften. Zwei- bis vierzeilige einfache rote Lom barden, selten mit kopfstempelförmigen Aussparungen. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, mit Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S., im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Neustift, Stiftsbibl., Cod. 158, Bl. Ir gehört Cod. 182 zu einer Gruppe von urspr. tw. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzel handschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`4 D 12 (Tinte, radiert), II 2 H 30 (Bleistift) und Nr. 182 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 182. Bl. 1r Cod. 183 209 Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol II 167, 208. – Stefani 106. – Peintner, Stiftsbibliothek 121. (Ir–v) leer. (1ra–16rb) ‹Ps.-›Eusebius Caesariensis: Epistola ad beatum Damasum episcopum Portuensem et ad christianissimum Theodosium Romanorum senatorem de morte Hieronymi (PL 22, 239‑282. Klapper II 10–241. – BHL Nr. 3866. BHM Nr. 903E). Abweichende Kapiteleinteilung. Schlussschrift mit Datierung: Explicit epistola beati Ewsebii ad Damasium Portuensem episcopum et ad Theodonium Romanorum senatorem de morte gloriosissimi confessoris Ieronimi doctoris eximii anno domini MoCCCCo in die sancti Remigii episcopi et confessoris etc. (1. Oktober 1400). (16va–b) nur Rahmung. (Ir–v) leer. P. A. Cod. 183 HENRICUS SUSO Pap. I, 48, I* Bl. 285 × 205. Süddeutscher Raum, um 1400. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: Neustift, Stiftsbibl., Cod. 83 (dat. 1400) und Cod. 158. Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + 4.VI48 + (I–1)I* (Nachsatzbl. 18. Jh.). Lage 2 und 3 am Lagenende Kustoden in arabischen Ziffern erhalten. Am oberen Blattrand mittig zeitgenössische Foliierung in Rot 2–38 (Bl. 1 ungezählt, Zahl 2 doppelt angeführt, 11 ausgelassen), dann Forts. 19–24 und 35–38, darüber Korrektur Foliierung 19. Jh. durch die ULB Tirol 39–48 (hier berücksichtigt). Am oberen rechten Blattrand Foliierung 15. Jh. in Schwarz 166–203. S: Schriftraum ca. 220 × 155. Zwei Spalten, von Tintenlinien gerahmt, zu 43–46 Zeilen. Bastarda. Am Blattrand vereinzelt Ergänzungen, Korrekturen und Kapitelzählung, tw. von gleicher, tw. von späterer Hand. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen und Überschriften. Zwei- bis vierzeilige einfache rote Lombarden (Bl. 1ra mit kopfstempelförmiger Aussparung und verziertem Ausläufer). Reprä sentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, mit Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. HOROL. SAPIENT., im dritten Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Nach dem Inhaltsverzeichnis in Neustift, Stiftsbibl., Cod. 158, Bl. Ir gehört Cod. 183 zu einer Gruppe von urspr. tw. selbständigen Hs. des 14. und 15. Jh., die im 15. Jh. in Neustift zu einem Sammelband vereinigt wurden (s. auch Foliierung). Im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. wurde die Sammelhandschrift (wie dies auch bei anderen Hs. und Inkunabeln gemacht wurde) in sieben Einzel handschriften aufgelöst; jeder Einzelband erhielt den für Neustift typischen Halbledereinband. Zur urspr. Hs. zählen folgende Signaturen (in der urspr. Reihenfolge): Cod. 158: Augustinus. Foliierung 15. Jh. 1–69. Cod. 83: Theologische Sammelhandschrift, dat. 1400. Foliierung 15. Jh. 70–128. Cod. 82: Henricus de Langenstein: De contractibus. De contemptu mundi. Foliierung 15. Jh. 129–165. Cod. 183: Heinrich Seuse: Horologium sapientiae. Foliierung 15. Jh. 166–203. Cod. 161: Iohannes de Dambach. Origenes. Foliierung 15. Jh. 204–215. Cod. 182: Ps.-Eusebius, dat. 1400. Foliierung 15. Jh. 216–231. Cod. 162: Sermones de dedicatione. Foliierung 15. Jh. 232–241. Von diesen sieben Hs. befindet sich heute eine an der ULB Tirol in Innsbruck (Cod. 82), die restlichen sechs werden hingegen in der Stiftsbibl. Neustift aufbewahrt. 210 Cod. 190 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II 4 D […] (Tinte, radiert), II 2 H 31 (Bleistift) und Nr. 183 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 183. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: P. Künzle (Hrsg.), Heinrich Seuse, Horologium sapientiae. Erste kritische Ausgabe unter Benützung der Vor arbeiten von Dominikus Planzer (Spicilegium Friburgense 23). Freiburg 1977, 162–163, Nr. 131. – K atalog ULB Tirol II 168, 208. – Kustatscher–Korenjak 154. (Ir–v) leer. (1ra–48vb) ‹Henricus Suso›: Horologium sapientiae (Ed. Künzle 363–605. – Kaeppeli Nr. 1852. Bloom field Nr. 5416). Tit.: Horologium sapiencie. (1ra) Prolog. – (2ra) Lib. 1. – (30vb) Lib. 2. (48vb) Schlussschrift: Explicit horologium sapiencie fratris cuiusdam ordinis Predicatorum etc. Darunter Tit.: Liber contemplacionis etc. (nicht auf den in dieser Hs. enthaltenen Text bezogen, wohl Tit. des urspr. darauffolgenden Textes in der ehem. Sammelhandschrift, möglicherweise Cod. 161 [Iohannes de Dambach, Tractatus de indulgentiis], oder eines urspr. vorgesehenen, nie ausgeführten Textes „De contemplatione“). (Ir–v) leer. P. A. Cod. 190 BRUNO DE SEGNI Perg. I, 68, I Bl. 315 × 250. Neustift (?), 2. H. 15. Jh./Anfang 16. Jh. B: Vereinzelt kleine Löcher und Nahtspuren. Bl. 17 Riss am seitlichen Blattrand. Die ersten drei Blätter am seitlichen und unteren Rand stark beschnitten (275 × 205, urspr. möglicherweise Rankendekor?). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + III6 + 7.IV62 + III68 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vorbzw. Nachsatzbl. am VD bzw. HD aufgeklebt. Urspr. wohl durchgehend Quaternionen: Bei der ersten und letzten Lage fehlt jeweils das äußere Doppelbl. (Textverlust), zudem nach Lage 1, 2 und 3 wohl jeweils eine Lage (Textverlust), nach Lage 4 mehrere Lagen verloren (Textverlust). Ab Lage 3 Reklamanten, Bl. 22v beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 245/250 × 175/180. Zwei Spalten, von tw. sehr feinen Tinten-, tw. Blindlinien gerahmt, zu einheitlich 44 Zeilen auf sehr feiner Tintenlinierung. Kalligraphische Bastarda. A: Rote Auszeichnungsstriche, Paragraphzeichen (selten blau), Unterstreichungen (u. a. Psalmen), Zeilenfüllsel und Überschriften; ab Bl. 67v keine Rubrizierung. Gelegentlich (in oberster Textzeile) cadellenartig ge dehnte Oberlängen (Bl. 42vb, 44va: größer und mit schwarzem Fleuronnée verziert). Ein- bis fünfzeilige Lombarden mit Punktverdickungen, einigen Aussparungen (Zickzack, Trifolien) und (kurzen) Ausläufern (zur Farbigkeit s. u.); Bl. 14rb nicht ausgeführt. Vereinzelt Repräsentanten. Die Anfänge der einzelnen Psalmen werden durch größere Initialen gekennzeichnet. Die Dekoration beginnt mit sechs aufwändigeren Fleuronnée-Initialen (vier- und fünfzeilig, Bl. 1ra, 2rb, 3ra, 4rb, 5rb, 6rb), gefolgt von sechs fein ausgeführten Ornamentalinitialen (fünfzeilig, Bl. 7rb, 8vb, 10ra, 11vb, 13ra, 14va), um sich bis zum Ende der Hs. – nur unterbrochen von zwei weiteren Ornamentalinitialen (sechs- und fünfzeilig; Bl. 19va, 38va) – mit einer Reihe einfach verzierter Lombarden zu begnügen. Fleuronnée-Initialen: rote und blaue Lombarden (Bl. 1ra rot-blau gespalten, Bl. 6rb graublau, wohl über malt, mit Einritzungen: Ranke?, VP-Ligatur) mit rot-grünem, oft in der Gegenfarbe gepunktetem Fleuron née. In den Binnenfeldern (tw. langstielige) Knospen in diversen Anordnungen, Bl. 2rb, 3ra und 4rb auf vollfarbigem grünem Grund (Grün dünn aufgetragen). Konturlinie(n), (angedeutete) Initialfelder, fallweise Zwickel-Trifolien. Als Besatz gereihte und zu Dreiergruppen zusammengefasste Knospen sowie Fibrillen; 211 Cod. 190 längere und kürzere Fadenausläufer, v. a. Bl. 1ra kammartig abstehend und dabei spitz zulaufend. Binnen feldknospen Bl. 1ra und 6rb erinnern etwas an das Fleuronnée in Sign. 14820 (1. Fünftel 16. Jh.); Bl. 6rb Besatzknospen, die mit zwei kurzen Parallelstrichen mit den Konturlinien verbunden sind; solche Knospen auch bei der Cadelle Bl. 44va, was darauf schließen lassen könnte, dass das Fleuronnée Bl. 1ra–6rb und das Cadellenfleuronnée einer Hand zuzuschreiben sind; letzteres orientiert sich wiederum am Fleuronnée der beiden Bände des Zollner-Graduales, stammt aber wohl nicht vom einschlägigen Hauptflorator, der noch akkurater zeichnete, vgl. z. B. Bl. 44va und die Cadelle Bl. 3v (oben) im ersten Gradualband. Auf dieser Basis ergibt sich – mit der gebotenen Vorsicht – eine Fleuronnée-Datierung in die 2. H. 15. Jh. (möglicher weise mit Tendenz ins 1. Fünftel 16. Jh.). Die rankenlosen und einfach gerahmten Deckfarbeninitialen Bl. 7rb–14va von einer Hand: Bl. 7rb Quer strich des A windet sich wie kurzes Spruchband (mit Schattenwurf auf kobaltblauem Initialfeld); Buchsta benkörper mit Füllranken, deren Blätter akkurat (mit Tendenz zur Symmetrie) ondulieren. Ungewöhnliche Initial- bzw. Binnenfeldgestaltungen: Bl. 8vb liegt der Buchstabenkörper auf einer „Blätterdecke“ auf, Bl. 10ra wird er von einem weiß-blauen Band umgeben, das an Wolkenstilisierungen erinnert, Bl. 11vb, 13ra und 14va zart und zugleich plastisch herausmodellierte Binnenfeldblüten und -blätter; zierliches „Erbsen schotenmotiv“ Bl. 10ra im Buchstabenkörper, 11vb in den Initialfeldblättern. Bis auf die (dunkel-)weinroten Buchstabenkörper Bl. 8vb und 10ra klare Farbigkeit. Bl. 8vb und 10ra Pinselgold im Binnenfeld. Bl. 19va und 38va wohl jeweils von anderer Hand: Bl. 19va sorgfältige Ausführung, gedeckte Farbigkeit (violettes Initialfeld mit blauer Rankenzier, auf dem der braunrot-ockergoldene Buchstabenkörper aufliegt; „Erbsen schotenmotiv“ in Füllblättern) und längere Akanthusranke; Bl. 38va blaue Lombarde mit Liniendekor (s. u.) umfangen von oliv-goldenem Initialfeld, das flüchtig in Gelb und Rot dekoriert ist (im Binnenfeld Netz muster). Die fünfzeiligen grünen Lombarden Bl. 18va und 20rb, in deren Buchstabenkörpern Blattdekor angelegt ist, evtl. für weitere Dekoration vorgesehen. Ab Bl. 16vb Lombarden in Blau, Weinrot und Grün (dünnerer Farbauftrag) mit einfachem, meist auf dem Buchstabenkörper aufgemaltem Dekor (in der Regel linear); Bl. 25vb fünfzeilige Lombarde: Buchstaben körper in pastosem Grün, roter Federzeichnungsdekor; Bl. 35ra sechszeilige abgeriebene Goldlombarde (Bolus sichtbar) mit rotem Fleuronnée (langstielige Knospen aus schraffiertem Binnenfeldgrund ausgespart, radial abstehende Fadenausläufer); diese Fleuronnéehand, die nicht mit dem am Beginn der Hs. tätigen Florator übereinstimmt, in flüchtigerer Manier auch Bl. 57ra. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. etwas abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck EXPOSIT. IN PSALMO DAVIDIO (!) in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot ge sprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signatur der ULB Tirol II 3 E 3 (Tinte, Bleistift) und Nr. 190/(1) (Tinte, Bleistift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 190/(1). Bl. 1r am unteren Blattrand bzw. Bl. 4r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r am unteren Blattrand bzw. Bl. 4r Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 1852 Suppl. – Katalog ULB Tirol II 180, 208–209. – Peintner, Stiftsbibliothek 121. 1 (Ir–v) leer. (1ra–67ra) ‹Bruno de Segni›: Expositio in Psalmos, unvollständig (PL 164, 699B–1228C. – Stegmüller RB Nr. 1852). Beginn des Textes fehlt, Bl. 1ra Beginn des zweiten Psalms. (1ra–6vb) Ps 2–7. Inc. mut. (PL 164, 699B): ] Et merito quidem contra eos cum quadam indigna cione loquitur qui in Christum dominum … – Expl. mut.: … qui omnia iudicas in equitate. Iudica me domine. Tu inquit me iudica [ (PL 164, 717C) (7ra–14vb) Ps 27–33. Inc. mut. (PL 164, 786A): ] et non edificabis eos. Isti exterminio digni sunt … – Expl. mut.: … Iste qui hoc dicit iam eius degustaverat [ (PL 164, 808A) 212 Cod. 194 (15ra–30vb) Ps 49–61. Inc. mut. (PL 164, 875A): ] ituri sunt qui semper celestia dilexerunt … – Expl. mut.: … dyabolum dereliqui veni ad Christum ipse [ (PL 164, 918A) (31ra–38vb) Ps 67–70. Inc. mut. (PL 164, 941B): ] erat cor unum et anima una et nemo dicebat aliquid … – Expl. mut.: … Quia dixerunt inimici mei mala mihi et qui custodiebant [ (PL 164, 964C) (39ra–62vb) Ps 117–143. Inc. mut. (PL 164, 1142B): ] non erit. Sed illi soli dicant qui dominum timent … – Expl. mut.: … ex hoc in illud quia eorum [ (PL 164, 1213D) (63ra–67ra) Ps 144–150. Inc. mut. (PL 164, 1216C): ] salvos faciet eos. Hoc est est (!) enim eorum voluntas … – Expl.: … Omnis spiritus laudet dominum, spiritus omnis eum benedicat nunc et in evum. Laudet ametque colat quem Rachel mater adorat. Nunc fecunda quidem sterilis sine semine pridem. Hoc pater hoc flamen hoc filius audiat. Amen. 2 (67rb–68vb) Dispositio catholicae fidei, unvollständig (vgl. Stegmüller RB Nr. 3339–3340, 9054, 9899). Tit.: Incipit disposicio katholice fidei quam edidit sanctus episcopus Athanasius in concilio Nicee civitatis coram multis patribus. – Inc.: Quicumque vult salvus esse ante omnia opus est ut teneat katholicam fidem … – Expl. mut.: … Tangere se permisit causa confirmandi katholicam fidem ut destrueret perfidiam hereticorum qui dicunt Christum non levasse carnem suam in celo et propter hoc Christus petivit cibum et ante discipulos manducavit expletis XL diebus coram discipulis [ Zu den Kommentaren zum Athanasianischen Glaubensbekenntnis vgl. A. E. Burn, The Athana sian Creed and its early Commentaries. Cambridge 1896. (Ir–v) leer. A. P. / U. S. Cod. 194 RITUALE NEOCELLENSE Perg. I, 67 Bl. 305 × 225. Neustift, 1507. B: Dickes Perg. Buchblock nachträglich leicht beschnitten (vgl. Zahlen am oberen Blattrand). Lagen: (V–2)I,7 + IV15 + (IV–2)23(21) + IV31(29) + (IV–1)38(36) + 3.IV62(60) + (IV–1)70(67). Gegenbl. zu Bl. I herausgeschnitten (ohne Textverlust). Gegenbl. zu Bl. 7 als Spiegel auf dem VD aufgeklebt. Gegenbl. zu Bl. 19 und 20 (urspr. Bl. 21 und 22) sowie zu Bl. 64 (urspr. Bl. 69) mit Textverlust, Gegenbl. zu Bl. 33 ohne Textverlust herausgeschnitten. Auf der Miniatur Bl. 70v am oberen und rechten Blattrand Abklatsch von Rubrizierung. Am oberen Blattrand zeitgenössische Foliierung in roten arabischen Ziffern 1–70; urspr. Bl. 20, 21 und 69 fehlen (herausgeschnit ten). Zwei Pergamentsignakel. S: Schriftraum 215 × 150/155. Von Blindlinien gerahmt, zu einheitlich 22 Zeilen auf Blindlinierung bzw. sieben rote Vierliniensysteme und Text. Textualis formata (Textura), dat. 1507 (Bl. 70v). Schreiber (?): S. S. (= Ste phan Stetner) (Bl. 70v), vgl. Krämer, Scriptores. A: Rote Auszeichnungsstriche und Überschriften; wenige blaue Paragraphzeichen. In den Binnenfeldern der Majuskeln gelegentlich feine Doppelschrägstriche, tw. mit einseitigem (bedorntem) Bogenbesatz (z. B. Bl. 2v, Z. 3). Rote und blaue Benediktionskreuze in Tatzenkreuzform. Zur Wasserweihe drei blaue Zeichen (jeweils eine Notenzeile hoch), die die schriftlichen Handlungsanweisungen visuell unterstützen (vgl. Sign. 14820): Bl. 38v Kreuz (mit dreiteilig aufgespaltenen Enden und weinrotem Federzeichnungsdekor) zu hic insuffla per modum crucis sancte, Bl. 40r vierstrahliger Stern mit Goldpunkt in der Mitte zu hic proijciatur aqua in quatuor partes, Bl. 41v dreiblättrige Lilie zu suffla ter in fontem in hunc modum. Ein- bis dreizeilige (dunkel-)rote und blaue, tw. verblasste Lombarden mit Punktverdickungen, in wenigen Fällen kopfstempelartiger Besatz und in gestrichelte Achterschlingen gelegte, mit freien Kreisen sowie Fibrillen dekorierte Ausläufer (sehr ähnlich in Cod. 85 und 405, vgl. z. B. Bl. 22r mit Cod. 85, Bl. 1r bzw. Cod. 405, Bl. 160v); Bl. 66v vierzeilige kopfstempelartig in Rot und Blau gespaltene Lombarde mit Fleuronnée (im Binnenfeld rot, außen blau; zur Einordnung s. u.). Repräsentanten. Cod. 194 213 Deckfarbenschmuck: 13 zwei- bis vierzeilige ornamentale Initialen mit kurzen Ranken, nur Bl. 1r1 achtzei lig mit annähernd dreiseitig umlaufender Ranke; zwei ganzseitige Miniaturen. Die Ornamentalinitialen (Bl. 1r1, 1r2, 1r3, 7v, 8r, 21v, 23r, 27r, 32v, 36r, 45v, 54v, 57r) mit einfachen Rah men, häufig in der Farbe der Außengrundzwickel, diese oft mit Trifolien gefüllt; Bl. 1r1 Rahmenleisten grün (oben und unten), ockerfarben (links) und violett (rechts). Bei blauen Buchstabenkörpern, auch bei der größeren Einleitungsinitiale, Blattfüllungen nur angedeutet, bei andersfarbigen Buchstabenkörpern Füllblätter plastischer herausgearbeitet. Binnenfelder meist blattvergoldet und punziert (Rautengitter, häufig mit siebenblättrigen Blüten gefüllt, s. auch Goldgrund Bl. Iv); in Deckfarbe ausgeführte Binnenfelder (Bl. 27r, 32v) mit gegenständigen Blattpaaren. Kurze, kräftig bewegte Akanthusranken (häufig Gabelungen, Überschneidungen und Verschlingungen), in der Regel aus Serifen der Buchstabenkörper entspringend, auch am Rahmen ansetzend; Blattformen wenig variiert, immer wieder tropfenförmig verdickte Blattadern; als Schmuck Goldfelder und -punkte (mit tintenfarbenem Federzeichnungsdekor); Bl. 1r Marginalranken zusammengesetzt aus zwei auf dem linken Seitenrand miteinander verschlungenen Hauptpartien (an der Verknüpfungsstelle hakt sich auch der Ausläufer der mittleren Initiale ein); große Fibrillen, links unten ein naturalistisch gestalteter Astanschnitt, üppige Fantasieblüten, auf der Spitze einer solchen steht (auf dem rechten Seitenrand) ein Storch mit weit nach hinten gebogenem Hals. Bl. Iv Rahmen (Breite 21–22 mm) der Miniatur aus zusammengesteckten Stäben, deren Zwischenräume bzw. -streifen mit Blattranken (auch hier verdickte Adern) gefüllt sind; äußere Rahmenstäbe gehen an den Ecken in kurze Ranken über (vgl. Ornamentalinitialen); mittig an den Leisten Goldpunkte mit weinrotem Federzeichnungsdekor. Etwas sorg fältiger (sowohl Initiale selbst als auch Rankenwerk): Bl. 54v und v. a. Bl. 57r, bei denen im Gegensatz zu dem etwas blasseren Grün, das bei allen anderen Initialen eingesetzt wurde, Olivgrün Verwendung fand; generell begrenzte Farbpalette; ockerfarbene Partien mit Pinselgold; Blattgold auf rotem Bolus (s. Bl. 57r). Bl. Iv (236 × 162) Madonna. Muttergottes mit opulenter Perlenkrone sitzt auf Holzthron; das dunkelrote Ehrentuch (mit Goldmuster, das Blätter und Früchte beinhaltet) reicht über die Sitzfläche bis auf den Boden hinunter und dient Maria als Bankbelag; in der Rechten hält sie eine kleinere grüne melonen- bzw. kürbisar tige Frucht, von der sich das Jesuskind, das Maria mit der linken Hand stützt, abzuwenden scheint; die Handfläche seines erhobenen, rechten Händchens weist nach vorne. Insbes. die Marienfigur unsicher propor tioniert: der rundliche Kopf im Verhältnis zum Körper zu groß, der sich unter den (allerdings gekonnt he rausgearbeiteten) Gewandfalten nicht recht abzeichnet, sodass die Sitzposition der Gottesmutter vage bleibt; der Thron detailreich (bis hin zur Maserung), aber perspektivisch etwas unsicher gestaltet (Fluchtlinien der Rückenlehne zu weit nach oben geführt). Bl. 70v (258 × 159) Hl. Augustinus. Halbfigur des Heiligen mit prächtiger Mitra, grünem Pluviale und gold farbenen, beringten Handschuhen präsentiert in einem fensteröffnungsartig gestalteten Rahmen vor rotem Teppichgrund (mit Goldranken) ein aufgeschlagenes Buch mit dem Incipit der Augustinusregel von drei Händen: 1) Hec sunt que ut observetis, 2) precipimus in monasterio constituti, 3) post p e k (principalia pre cepta); Gesichtszüge durch kräftige schwarze Linien bezeichnet (Überarbeitung?); in die Rahmenkehlungen „Gewändefiguren“ eingestellt (auf zierlichen Säulchen, in Grisaille-Malerei): zwei bis auf ihr Lendentuch nackte, bärtige Männer; der linke hält ein Schwert über seinen Kopf, der rechte einen Stein; der Heilige an den rechten Rand der Fensteröffnung gerückt, wobei er das Buch, das nicht auf seiner linken Handfläche auf liegt, sondern darüber zu schweben scheint, in seine linke Armbeuge bettet und mit dem rechten Zeigefinger hält; auf dem Vorderrand der Fensterbrüstung Jahreszahl 1507 eingemeißelt (flankiert von Dreiblättchen), rechts davon Signatur S S (= Stephan Stetner; nach dem Buchstaben jeweils charakteristisches Ziermotiv bzw. Trennzeichen aus drei kleinen Kreisen in Dreiecksanordnung mit kurzem, kommaartigem Fadenaus läufer); über dem Kopf des Augustinus üppiges, rot liniertes und in Achterschlingen gelegtes Schriftband mit Datierung Anno domini millesimo quingentesimo septimo (mehrfach eingestreutes Ziermotiv: Punkt bzw. kleines Quadrat, besetzt mit Häkchen und Fibrillen). Das Fleuronnée mit länglich-blasigen Knospen und (bedornten) Besatzknospen, die in zwei kürzere Fäden auslaufen, von der im 1. Fünftel 16. Jh. nachweisbaren Hand, die auch in Cod. 405 (dat. 1501, hier das Fleu ronnée flüchtiger) und Cod. 142, T. I (dat. 1521) tätig war, in beiden Bänden des Zollner-Graduales einige Initialen mit Fleuronnée (s. jeweils Ergänzungshand 2) geschmückt und Sign. 14820 zur Gänze ausgestattet hat. Diese Hand ist vermutlich auch für den ornamentalen Deckfarbendekor (einschließlich Rahmen und Ranken Bl. 1v) in der vorliegenden Hs. sowie in Cod. 405 (hier nicht so sorgfältig ausgeführt) verantwort lich. Detailliert zu dieser Hand und den Zusammenhängen zw. Fleuronnée- und Deckfarbenzier bei Sign. 14820. Die beiden sorgfältiger ausgearbeiteten Initialen Bl. 54v und 57r zeigen Affinitäten zur Epistolar hand von Cod. 196, Bl. 1r, 2r und 7v: Blattwerk im Buchstabenkörper ebenfalls mit kleinen Randeinbuch tungen und tropfenförmig verdickten Adern, Bl. 2r zudem auch das Binnenfeld mit blütengefülltem Rauten gitter punziert; darüber hinaus auch Ähnlichkeiten im Rankenwerk (Blattformen, Überschneidungen). Bislang ging man aufgrund der Signatur Stephan Stetners in der Miniatur Bl. 70v davon aus, dass er nicht nur als Schreiber (vgl. Cod. 100 und möglicherweise auch Cod. 196), sondern auch als Buchmaler tätig ge wesen sei (s. z. B. Hermann 170–171, Fiegl, Prachthandschrift 748), was an dieser Stelle nicht völlig negiert, 214 Cod. 194 aber mit der gebotenen Vorsicht erwogen werden soll. Denn in den von ihm erhaltenen Werken hat Stetner nur in Cod. 194 innerhalb einer Illumination signiert, ansonsten jeweils im Text (s. die vier Signaturen in Cod. 100). Zudem gehört seine Signatur in Cod. 194 nicht urspr. zur Jahreszahl auf der Fensterbrüstung, sondern wurde dahinter und damit zugleich unterhalb der Augustinusregel ergänzt, womit ein Bezug zum geschriebenen Text, also zu seiner Schreibertätigkeit, ausgedrückt sein könnte. Insbesondere die Augustinus-Miniatur, deren unlängst vorgeschlagene Zuschreibung an Friedrich Pacher (s. Suckale, Suckale-Redlefsen 60–61) angezweifelt werden muss, zeigt einige Ähnlichkeiten zum Wappen halterengel in Cod. 405; darüber hinaus lässt sich der hl. Augustinus gut mit der Figur des Reginbert in der Miniatur am Anfang der Neustifter Totenrotel von 1515 vergleichen (s. H. Miekisch, Das Augustiner-Chor herrenstift Neunkirchen am Brand. Münsterschwarzach 2014, Farbabb. S. 170). S. R. E: Renaissanceeinband: helles Schweinsleder über Holz mit Blinddruck. Neustift (?), 2. H. 16. Jh. Leder an den Stellen der einstigen Beschläge und in den Ecken beschädigt, am HD etwas abgerieben. VD und HD annähernd gleich: drei von jeweils dreifachen (außen sechsfachen) Streicheisenlinien begrenzte Rahmen. Rahmen 1: verschränkte Bögen mit floralem Dekor (Nr. 1 = ähnlich wie Neuhauser, Rolleneinbän de, R 10), seitlich je eine Rolle, unten und oben je zwei spiegelsymmetrisch angeordnet. Rahmen 2: zu sätzlich vertikal mit dreifachen Streicheisenlinien unterteilt, Feld leer, Rahmen gefüllt mit Rolle Sichel mit eingeschriebenem Blattmotiv abwechselnd mit floralem Dekor (Nr. 2). Rahmen 3: Rolle mit Kornblumen motiv (Nr. 3). Spuren von vier rautenförmigen Eck- und einem viereckigen, auf die Spitze gestellten Mittel beschlag. Zwei Kurzriemen-Leder-Metall-Schließen. Kanten gerade und nach innen abgeschrägt. Rücken: blau gefärbt (wie öfters in Neustift), vier Doppelbünde. Bünde von zwei- und dreifachen Streicheisenlinien begrenzt, Linien auf VD bzw. HD fortgesetzt, spitz aufeinander zulaufend und leicht in den ersten Rahmen hineinreichend. Im ersten Feld Rest eines aufgeklebten Papierschildes 1507, im zweiten Rest eines Papier titelschilds R[it]ua[le] / [C]o[ll]egi[i] / Neocel., beide vor der Blaufärbung des Rückens aufgeklebt, im dritten Signaturschild (s. bei G). Kapitale mit naturfarbenem und hellbraunem Spagat umstochen. Schnitt abwech selnd rot und grün gesprenkelt. Spiegel des VD Pergamentblatt mit Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leeres Papierblatt. Unter den Spiegeln zusätzliches Blatt aufgeklebt, Gegenfalz (mit Resten einer Rahmung) zw. Lage 1 und 2 bzw. 8 und 9. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 3 E 7 und N. 194 sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 194. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italie nischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. Ir Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm II 110. – Hermann 170–171, Nr. 181. – Atz 1012. – Colophons Nr. 16895. – P. d’Ancona, E. Aeschlimann, Dictionnaire des miniaturistes. Milan 1949, ND Nendeln 1969, 200. – Gschwend 86–88 (ohne Nennung der Sign.). – Lipphardt III 1084–1086; VI 352, Nr. 628 (Sigle Neust3). – Neuhauser, Neustift 85. – Katalog ULB Tirol II 193, 209. – Katalog ULB Tirol II, Beiheft, Taf. 77–78. – Peintner, Stiftsbibliothek 126. – Gozzi 548, Nr. 79. – R. Kay, Pontificalia. A Repertory of Latin Manuscript Pontificals and Bene dictionals. Lawrence 2007, 108, Nr. 560. – Baroffio 324, Nr. 16714. – Krämer, Scriptores, s. v. „Stetner, S.“. – R. Suckale, G. Suckale-Redlefsen, Buchmalerei in Mitteleuropa und ihr historischer Kontext, in: J. F. Hamburger, R. Suckale, G. Suckale-Redlefsen (Hrsg.), Unter Druck. Mitteleuropäische Buchmalerei im Zeitalter Gutenbergs (Buchmalerei des 15. Jahrhunderts in Mitteleuropa 2). Luzern 2015, 11–66, hier 60–61. – U. Stampfer, Neustifter Rituale, in: J. Kronbichler, H.-P. Ties (Hrsg.), Renaissance im Gebirge. Der Maler Bartlme Dill Riemenschneider und seine Zeit. Hofburg Brixen 27. Mai–31. Oktober 2017, 118–119 (Kat. 25). Rituale Neocellense. Vgl. Cod. 726 und Sign. 14820. (Ir) leer bis auf Stempel (s. bei G). (Iv) ganzseitige Miniatur (s. bei A). (1r–7v) In purificatione BMV. (1r) Benedictio candelarum. Tit.: Purificacionis Marie cereorum benedictio post 3as. – (6r) Processio candelarum. – (6v) Canticum Zachariae. (7v–19v) Dnca in palmis. (7v) In die palmarum ad processionem. – (10r) Exorcismus florum et frondium. – Benedictio pal Cod. 196 215 marum. – (14r) Praefatio. – (16r) Aspersio et thurificatio palmarum. – (16v) Distributio palmarum. – (17r) In reditu. – (18r) Adoratio crucis. – (19r) Canticum puerorum. – (19v) In reditu ante altare oratio. (19v–23r) Fer. 5. in coena dni. (19v) In cena domini benedictio ignis. – Expl. mut.: … nostris profuturum usibus novum hunc ignem sanctifica [ Bl. 20 und 21 herausgeschnitten, enthaltend Fußwaschung. Bl. 22r Inc. mut.: hu]militatis gratia Petri petens vestigia … (23r–32r) Fer. 6. in parasceve. (23r) Adoratio crucis. – (26r) Depositio crucis (Gschwend 86). – (27r) Vesper. (32v–42r) In vigilia paschatis. (32v) Benedictio ignis. – (34v) In reditu canitur himnus Prudencii (AH 50, Nr. 31). – (35r) Exsultet (nur Verweis). – (35r) Benedictio aquae. – (42r) Litaniae sanctorum (nur Verweis). (42v–45r) Dnca resurrectionis dni (Gschwend 86–88). (45r) Benedictio aquae in vigilia Pentecostes (nur Verweis). (45v) In assumptione BMV. (45v–54v) Ordo paenitentiae. (54v–56v) Einkleidung der Novizen. Tit.: De induitione (i gestrichen) novitiorum. (57r–64r) Profess. (64v–66r) Ordo für die Aufnahme von Laien. Tit.: De fraternitate laicali. (66r–68v) Totenliturgie für die Konventualen. (68v) Expl. mut.: … Infra sacerdotium constituti tria orabunt psalteria cum septem vigiliis. Conversi vero trecen[ta Bl. 69 herausgeschnitten (mit Schluss des Textes), s. bei B. (70r) nur Rahmung und Foliierung. (70v) ganzseitige Miniatur (s. bei A) mit zweimaliger Datierung Anno domini millesimo quingentesimo septimo (Spruchband) sowie 1507 (untere Stirnseite des Rahmens) und Schreibernennung (?) S. S. W. N. / U. S. Cod. 196 LECTIONARIUM MISSAE NEOCELLENSE Perg. I, 26 Bl. 315 × 250. Neustift, Ende 15. Jh. (vor 1482?). B: Perg. am unteren Blattrand stark abgegriffen. Lagenaufbau im Zuge der Neubindung in der 2. H. 18. Jh. verändert, urspr. wohl durchgehend Quaternionen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 2.IV16 + (IV–4)20 + (VI–10)32(23) + II35(26). Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Gegenbl. zu Bl. 17–20 herausgeschnitten (wohl leer, ohne Textverlust). Von urspr. Sexternio Gegenbl. zu Bl. 31 im Jahre 1746 he rausgeschnitten, heute in der Kathedralbibliothek Esztergom (Ungarn) (s. bei G). Bl. 21–29 sowie Bl. 36–42 1872 an der ULB Tirol herausgeschnitten (Textverlust), s. bei G. Bl. 30 und 31 als Einzelbl. mit der letzten Lage verbunden. Am oberen Blattrand Foliierung durch die ULB Tirol 1857 (vgl. Innsbruck, ULB Tirol, Alte Titelkopie: Handschriften). Bl. 32[23]r Spur eines Ledersignakels sowie am Blattrand römisches Zahl zeichen I (als Orientierungshilfe?). 216 Cod. 196 S: Schriftraum 210/215 × 145/155. Von feinen Tintenlinien gerahmt, zu einheitlich 19 Zeilen auf feiner Tinten linierung. Zirkelstiche. Textualis formata (Textura) Ende 15. Jh. (Terminus ante quem 1482?: vgl. Eszter gom, Kathedralbibl., Ms. II.2, Bl. 5v, figürliche Initiale dat. 1482). Vereinzelt mit Bleistift Akzentzeichnun gen (Terminus post quem 1746) und Korrekturen, Durchstreichungen. A: Rote Auszeichnungsstriche, Überschriften, Akzentzeichnungen (bis Bl. 5v Striche und Wellen, Bl. 32[23]r und 33[24]r auch Häkchen) sowie Verweiszeichen (z. B. Bl. 30[21]r; Korrekturen dieser Art könnten 1531 vorgenommen worden sein, vgl. ebda die entsprechende Datierung in Rot auf der unteren Initialrahmen leiste; Ziffern der Jahreszahl mit Zwischenpunkten geschrieben, als „Zeilenfüllsel“ Doppelpunkt mit kurzer Wellenlinie); Bl. 34[25]r rote Unterstreichung; Bl. 15r jüngere dunkelrote Ranke als Zeilenfüllsel über radierter Stelle. Bl. 3r, Z. 4 und 6 schwarze Minuskeln mit roten Majuskeln überschrieben; Bl. 14v in der obersten Zeile einzeilige rote Lombarde und nachfolgendes (blassrotes) Paragraphzeichen über radierter Stelle ergänzt; von dieser späteren Hand wohl auch die Füllranke Bl. 15r sowie ab Bl. 10v immer wieder Akzentzeichnungen in blassem Rot. In den Binnenfeldern der Textualis-Majuskeln des Öfteren feine Dop pelschrägstriche (z. B. Bl. 3r, Z. 1), fallweise auch mit kleinem angesetztem Bogen (z. B. Bl. 15r, Z. 2). Deckfarbenschmuck: 33 ornamentale und acht figürliche Initialen, jeweils mit Ranken; die ornamentalen Initialen vier- bis sechszeilig, die figürlichen fünf- und sechszeilig. Initialrahmen zumeist profiliert und rosafarben (auch blau) oder (bei etlichen figürlichen Initialen) mit Blatt gold belegt und punziert; einige I-Initialen rahmenlos, die mit den in der Längsschaftmitte symmetrisch angeordneten Blattknollen wie doppelflügelige Türen mit Knäufen wirken (z. B. Bl. 19v); Großteil der Ini tialfelder bzw. Außengründe mit punziertem Blattgold gefüllt; Bl. 3r und 4v silberne Initialfelder (bemalt bzw. punziert); zur Punzierung s. u.; Binnenfelder ansonsten meist blau oder karminrot ausgemalt und von feinen, gestrichelten Goldranken oder Rautengründen, Bl. 2v von maßwerkartigen Motiven in Weiß und Silber, überzogen. Die in Grün, Pinselgold, häufig Blau und Rosa, Rot und Violetttönen, seltener in GrauWeiß gehaltenen und meist von einem dünnen Konturstab eingefassten Buchstabenkörper, die mit vielfäl tigst variierendem Blattwerk gefüllt sind, liegen auf den Rahmen auf (Bl. 30[21]r–v der Binnenfeldgrund bis in die Rahmenzone hinein ausgedehnt); die Serifen tw. wie angeschnittene Pflanzenstängel gestaltet (z. B. Bl. 2r), vgl. Ranken; Bl. 5r Buchstabenkörper mit drei Goldringen geschmückt; der weiße Buchstabenkörper Bl. 14v wohl unvollendet. Die Marginalranken bei Ornamentalinitialen meist zwei- oder dreiseitig (in Sei tenecken oft in Schlingen gelegt); bei figürlichen Initialen, denen in der Regel aufwändigerer Marginaldekor zugeordnet ist, überwiegen kürzere Rankenarrangements; insgesamt filigran-anmutige Rankenanlage ohne Bestrebung, die Seitenränder abzudecken; die Äste dabei auf längeren Abschnitten schnurgerade geführt, Bl. 3r grüner und rosafarbener Ast zu einem Rankenstab gewunden; zwei Grundtypen des Randdekors: stilisierte Ranken (mit abwechslungsreichen, schmalsilhouettigen Blattvarianten, zur Farbgebung s. u., und sparsam, gerne mittig auf dem Bas-de-page platzierten Fantasieblüten) und botanisch identifizierbare Blü tenzweige (z. B. Bl. 8r Veilchen, Bl. 10v1 Iris, Bl. 12v Kornblume); die beiden Typen oft miteinander kom biniert (Charakteristikum), naturalistische Zweige häufiger auf Folios mit figürlichen Initialen; bei beiden Typen sowohl naturalistisch anmutende Astanschnitte (z. B. Bl. 2r) als auch dezenter, fallweise punzierter Golddekor: Punkte (selten mit bunten Blütenblättern besetzt, z. B. Bl. 4v), Tropfen und charakteristische (spindelförmige) Füllfelder in Rankenzwischenräumen (z. B. Bl. 2r linker Rand); Bl. 12r rosafarbener Dis telzweig mit naturalistischen Blüten und stilisierten Blättern (die ihrerseits an bestacheltes Distelblattwerk erinnern). Variationsreichtum auch beim Rankenansatz: Während im Epistolar die Ranken häufig aus den Buchstabenserifen oder aus (Blatt-)Manschetten (z. B. Bl. 8r), aber auch aus dem Rahmen (bzw. Initialfeld) entspringen, an diesem angelegt (z. B. Bl. 6v) oder sogar angebunden (Bl. 2r) sein können, bleibt die Rand zier im Evangelistar zuweilen ganz ohne Kontakt zur Initiale (z. B. Bl. 34v). Große Vielfalt der insges. ca. 90 Drolerien: häufig Vögel, zuweilen Insekten, z. B. Fliege (Bl. 5v neben Initiale), Biene (Bl. 14v Bas-de- page), Käfer (Bl. 18r oberer Seitenrand), auch Raupen, zudem lebendig inszenierte Vierbeiner, wie z. B. eine Katze, die einer Maus hinterherjagt (Bl. 14r), ein sich kratzender Hund (Bl. 12r) oder ein kauernder Hase (Bl. 9v); v. a. die Vögel wirken in Farbgebung und Haltung sehr naturnah, z. B. der Erpel, Bl. 19v, oder die Henne mit ihren Küken, Bl. 32r; zu Beginn der Hs. einige Vögel mit weit ausschreitender bzw. gegrätschter Beinposition (Bl. 2r–v, 3v); Bl. 10v auch das Osterlamm mit Siegesfahne drolerieartig in das Rankenwerk integriert. Figuren anatomisch recht gekonnt ausgeführt; beim stehenden Jesuskind Bl. 2r der Versuch eines Kon traposts, wobei der Standbeinfuß im rechten Winkel nach außen gedreht und der Spielbeinfuß spitz nach vorne gestreckt ist, vgl. die Christusfiguren Ms. II.2, Bl. 9r und v. a. Bl. 13v; hl. Michael (Bl. 33[24]r) mit ausgeprägtem Körperschwung. In der Regel rundliche Köpfe mit rosigen Pausbacken, woraus sich eine kindliche Physiognomie ergibt; Weißhöhungen auf Stirn, Augendeckeln, oberen Wangenpartien und Nasen rücken (zuweilen auch auf den Fingern); die Chorherren Bl. 31[22]r durch (leicht) variierende Gesichtszüge und unterschiedliche Haarfarben als jüngere und ältere Männer individualisiert. Gewänder zuweilen recht gebauscht und mit feiner Strichelung schraffiert und gehöht (vgl. Strichführung bei Buchstabenkörpern, Cod. 196 217 s. Bl. 32[23]r: Kleider, Decke, Vorhang und Buchstabenkörperfüllung); fallweise auch Goldhöhungen auf Gewandstoffen (z. B. Bl. 30[21]r). Einzelfiguren (Jesuskind, Bl. 2r; hl. Michael, Bl. 33[24]r; s. auch hl. Agnes, Ms. II.2, Bl. 4v) stehen auf grün marmorierten Böden (beim Jesuskind könnte es sich auch um ein Kissen mit ähnlichem Muster handeln) und werden von ornamentalen Gründen hinterfangen; szenische Darstellungen finden in Räumen (z. B. Huldigung an den hl. Augustinus, Bl. 31[22]r) oder Landschaften (z. B. Heimsuchung, Bl. 30[21]r) statt; die himmlische Sphäre der Allerheiligen-Darstellung (Bl. 34[25]v) durch stilisierte blau-goldene Wolkenbänder wiedergegeben; Wände (aus grau-grün und violett kolorierten Mauersteinen) tw. mit detailliert gemusterten Teppichen verhängt (z. B. Bl. 32[23]r) und Böden mit ge musterten Fliesen belegt, z. B. Dekor aus Kreuzen und Kreisen, Bl. 31[22]r und Ms. II.2, Bl. 13v, oder Adler (Tiroler Adler?), Bl. 32[23]r; Fensterscheiben mit Silber, Scheibenstruktur durch Rauten aus Punktpunzen linien bezeichnet (z. B. 31[22]r, Ms. II.2, Bl. 5v); typisch bei den Landschaftsdarstellungen die Bäume mit dünnen Stämmen und summarisch zusammengefassten Blattwerkpartien (z. B. Bl. 30[21]r; Ms. II.2, Bl. 4r); auch hier naturalistische Pflanzendarstellungen (z. B. Bl. 30[21]r; Ms. II.2, Bl. 1r, 2r). Bei Architekturen bzw. Innenräumen (bis auf die Maueraufsicht Bl. 34[25]r und das Kopfteil des Bettes Bl. 32[23]r) recht sicherer Einsatz von Parallel- und Zentralperspektive. Während die Farbgebung der Buchstabenkörper auch kräftiges Rot und Blau kennt, wurde bei den Ranken zarteres, fast transparent wirkendes Kolorit verwendet: dezentes Grün, helleres Mittelblau, Rosa (ganz selten Rot), Ocker und ins Fliederfarbene tendierendes Violett; verdunkelte Blattspitzen, an den Graten zarte Farb verläufe, auch Changieren möglich (Bl. 9v Blätter neben Initiale: gebrochenes Grün zu Altrosa); gekonnt herausgearbeitete Rundungen durch aufgesetzte Lichter, s. z. B. die Pollen der Fantasieblüte Bl. 7v. Reich liche Anwendung von Gold: Pinselgold patzig aufgetragen, Initialfeld Bl. 7v mit bes. grobem, möglicher weise nachgetragenem Pinselgolddekor in dachziegelartigen, schwarz konturierten Einzelfeldern, die zu sätzlich mit bunten Punkten geschmückt sind; Blattgoldpartien gut erhalten, an den seltenen Bruchstellen (z. B. Bl. 14v) zartrosafarbener Bolus sichtbar; Bl. 30[21]r Blattgold des Heiligenscheins samt Bolus sowie Teile der Hintergrund-Bemalung abgeplatzt, Vorzeichnung sichtbar. Aufwändigere Punzierung: nicht nur Punktlinien (radial als Dekor von Heiligenscheinen und Außengrundzwickeln) oder einfache Kreise, s ondern z. B. auch Blüte mit fünf spitzen Blättchen (z. B. Bl. 30[21]r und Ms. II.2, Bl. 6v, jeweils auf dem Initial rahmen), fünfblättrige Blüte mit gerundeten Blättchen in dachziegelartig aneinander gesetzten Punktpunzen bögen (Binnenfeld Bl. 13v) und gezahntes, von je zwei siebenblättrigen Blüten flankiertes Blättchen in dop pellinigem Rautengitter (z. B. Binnenfeld Bl. 9v); auch Kombination von Punzen und kleinteiliger Bemalung möglich, z. B. Binnenfeld Bl. 10v1 (bunte Blüten) und Initialrahmen Bl. 5v (blau-weiß-rote Querstriche). Bl. 2r Nacktes (beschnittenes) Jesuskind mit Heiligenschein auf grüner Bodenandeutung (Kissen?) stehend. Bl. 30[21]r Heimsuchung: Maria mit Heiligenschein und Elisabeth mit weißer Haube in hügeliger Land schaft. Bl. 30[21]v Marientod: Maria, im Kreis der Apostel im Bett sitzend, bekommt von Johannes die Sterbekerze gereicht, daneben Petrus mit Weihwasserkessel und Aspergil; über der Szene (auf einer Wolkenkonsole) die Halbfigur Christi mit dem Seelenfigürchen Mariens. Bl. 31[22]r Hl. Augustinus in prächtigem Bischofsornat, frontal thronend und die Rechte segnend erhoben, wird von adorierenden Augustiner Chorherren flankiert. Die beiden vorderen Chorherren halten ihre schwar zen Kappen in den Händen, links der Propst mit grauem, schweifbesetztem Feh-Almutium. Bl. 32[23]r Geburt Mariens: Anna sitzt im Bett, das durch einen grünen Vorhang verborgen werden kann, und übergibt Maria (Wickelkind) einer Magd; vor dem Bett ein mit Wasser gefüllter Zuber, ein Schemel und ein weißes Hündchen. Bl. 33[24]r Erzengel Michael als Seelenwäger mit erhobenem Schwert; in der linken Waagschale ein beten des Seelenfigürchen, an die rechte Schale klammert sich ein schwarzer Teufel. Bl. 34[25]r Kirchenweihe: Vor einem schmalen Rundbogentor (mit geöffneter Holztür und Vordach, darüber Rundfenster; grüne Dachziegel) Bischof im Ornat mit goldenem Krummstab (mit transparentem, wie Vor zeichnung wirkendem Sudarium) und Aspergil; hinter ihm zwei Kleriker mit Weihwasserkessel, Buch und Weihrauchfass; über der Szene weht die Kirchweihfahne (Kreuzfahne, vgl. Fahne des Auferstandenen Ms. II.2, Bl. 9r). Bl. 34[25]v Allerheiligen: Frontal thronender und von zwei Leuchter(?)-Engeln flankierter Christus mit Kreuzstab und Weltkugel, umgeben von Maria und Heiligen, die durch Haartracht und Kopfbedeckung nach (geistlichen) Ständen differenziert sind. Epistolar (Cod. 196, Bl. 1r–20v) und Evangelistar (Esztergom, Kathedralbibl., Ms. II.2, und Cod. 196, Bl. 30r–35[26]v, s. bei G und Inhalt) wurden jeweils vorrangig von einer Hand ausgestattet: Die sog. Evan gelistarhand ist als die versiertere und daher auch für die Seiten mit figürlichen Initialen zuständige Hand auszumachen; von ihr könnte auch das Jesusfigürchen im Epistolar (Cod. 196, Bl. 2r) stammen. Bei der sog. Epistolarhand handelt es sich um einen v. a. für die Folios mit Ornamentalinitialen zuständigen Mitarbeiter, der sich stark am Formenvokabular der Evangelistarhand orientierte, wodurch sich die Unterscheidung der beiden Kräfte fallweise etwas problematisch gestaltet; allerdings dürften die ornamentalen Hintergründe und 218 Cod. 196 die Punzierungen durchwegs von der Epistolarhand stammen. In aller Regel war jede der beiden Hände aber nicht nur mit der Initiale, sondern auch mit dem weiteren Buchschmuck auf dem jeweiligen Folio befasst, gestaltete also auch die Buchstabenkörper und das Rankenwerk samt Drolerien, vgl. z. B. die Buchstaben körperfüllungen der figürlichen Initialen Cod. 196, Bl. 2r, 33[24]r und Ms. II.2, Bl. 12v (alle Evangelistar hand), bei denen die Höhungen durch feine Strichelung herausmodelliert wurden, mit der Buchstaben körperfüllung der Ornamentalinitiale Cod. 196, Bl. 1r (Epistolarhand), die sehr ähnliche Formen (v. a. die tropfenförmig verdickten Adern), aber eine gröbere Ausführung zeigt, oder die Irisstängel Cod. 196, Bl. 10v (Epistolarhand) und Ms. II.2, Bl. 5v (Evangelistarhand) bzw. die Hasen-Drolerien Cod. 196, Bl. 9v (Epistolarhand) und Ms. II.2, Bl. 2v (Evangelistarhand), die jeweils große Motivnähe aufweisen, aber unter schiedlich gestaltet wurden. Man kann demnach davon ausgehen, dass es einen gemeinsam benutzten Vor lagenfundus (ein Musterbuch?) gab. Die Jahreszahl 1482 auf Bl. 5v von Ms. II.2 kann zusammen mit dem 1480 datierten Brevier Brixen, Priesterseminarbibl., Cod. T.3, das starke Bezüge zu Cod. 196 zeigt, wohl als zeitlicher Anhaltspunkt für die Entstehung beider Handschriftenteile herangezogen werden, die Datierung 1531 auf Bl. 30[21]r könnte sich auf Textkorrekturen beziehen (s. o. beim niederrangigen Buchschmuck), keinesfalls auf die Wirkungszeit der noch in der spätmittelalterlichen Buchmalerei verankerten Evangelistar hand (Roland, Renaissance 635 und 640 Anm. 25: 1531 als Datierung für die gesamte Hs. angenommen). Denn wäre das Evangelistar in dieser Zeit entstanden, wiese es vermutlich Verbindungen zu Cod. 100, dem sog. Posch-Missale (1524–1526), auf, in dem interessanterweise Motive aus Cod. 196 aufgegriffen werden (s. z. B. Iris Cod. 100, Bl. 68v). Die Evangelistarhand zeigt immer wieder Unterschiede in der Ausführung von Figuren (vgl. z. B. die nicht nur in Bezug auf Gesichter, Hände und Gewänder, sondern auch bei der Dar stellung von Tieren und Architektur bes. sorgfältige Einleitungsseite des Evangelistars, Ms. II.2, Bl. 1r, mit der Kirchenweihe, Cod. 196, Bl. 34[25]r). Inwieweit hierfür die das Gesamtkonzept der Hs. beherrschende Variationsfreude oder Qualitätsschwankungen verantwortlich sind oder ob man für diesen Handschriftenteil noch weitere Händescheidungen vornehmen muss, wird an anderer Stelle zu entscheiden sein. Der qualitätvolle Dekor von Cod. 196 und Esztergom, Kathedralbibl., Ms. II.2, weist deutliche maltechni sche und motivische Bezüge zu der Inkunabelbibel Wien, ÖNB, Ink. 3.C.6 auf (Straßburg: Johann Mentelin, vor dem 27. Juni 1466 bzw. um 1470; GW 4295 [mit Volldigitalisat]; zu dieser Bibel s. auch Roland, Gotik 291–292, Kat. 188 sowie Brixen, Priesterseminarbibl., Cod. C.13 und E.2), die ihrerseits durch Ranken mit tropfenförmig verdickten Blattadern und ihre naturalistischen Tierdrolerien und Blütenzweige bzw. -sträuß chen in der weiteren Nachfolge des wohl in der 2. H. der 1450er Jahre entstandenen Missales der Stiftsbibl. Innichen stehen dürfte (ohne Sign.; Hermann 75–78, Nr. 81; E. Kühebacher, Kirche und Museum des Stiftes Innichen. Bozen 1993, Nr. 163, Abb. 116, 117a–g; Roland, Gotik 275 Anm. 79, Abb. 17). Beispiele für Affinitäten zw. der Wiener Inkunabel und dem vorliegenden Lektionar: luzid-delikater Farbauftrag, durch aufgesetzte Lichter gekonnt herausgearbeitete Rundungen (s. z. B. Ink. 3.C.6, Bl. 176v die Beeren unten links oder Bl. 159r die Fantasieblüte auf dem rechten Seitenrand mit dem aus vielen Kügelchen zusammen gesetzten Fruchtkolben und dem „Erbsenschotenmotiv“ in den Blütenblättern; dieses Motiv übrigens nur im Evangelistar und dort auch nur einmal verwendet: Ms. II.2, Bl. 2r im Buchstabenkörper), Blätter mit ver dickter Ader und (dreiteiligen) Spitzen (z. B. Bl. 53r Bas-de-page), spindelförmige, blattvergoldete Ranken zwischenräume (s. ebda) sowie naturalistische Blütenzweige (z. B. Bl. 177r Heckenrosen und Veilchen) und Vogeldrolerien (z. B. ebda Eule und Falke). Figuren und Drolerien in der Inkunabelbibel allerdings wesent lich elaborierter, zudem die Kombination von naturalistischen und stilisierten Elementen in Cod. 196 und Ms. II.2 bis hin zum Charakteristikum ausgeprägt. S. R. E: Neuzeitlicher Bibliothekseinband: braunes Leder über Pappe. Neustift, 2. H. 18. Jh. Leder abgerieben und zerkratzt, in den Ecken tw. beschädigt, am VD links unten runder Fleck. Rücken: braunes Leder, vier einfache Bünde, von heute kaum mehr sichtbaren Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten EPIST. ET [EVA]N. in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh., am Spiegel des VD Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer bis auf Bleistiftnotiz 19./20. Jh. 1470 (Versuch einer Datierung?). G: Der Beginn des Evangelistars (Nativitas dni bis Petrus et Paulus, insges. 16 Blätter, 2 davon verloren) ge langte 1746 in den Besitz des Samuel Székely, vgl. Notiz Esztergom, Kathedralbibl., Ms. II.2, Bl. 1r: Ex cimeliis Sam[uel] Székely de Doba cui in Collegio Neocellensi Canonic. Regular. S. Augustini Congreg. Dioecesis Brixinensis in Tyroli d. 24. Martii 1746 per Rev. Casp. Mayer Bibliothecarium d[atum] est.“ Über Sándor Mérey, den Obergespan des Komitats Somogy, kam das Fragment 1847 schließlich in die Kathedral bibliothek von Esztergom (vgl. Hinweis Ms. II. 2, Bl. Ir). In diesem Teil Bl. 5v die Datierung 1482 (in Cod. 196, Bl. 30r, zusätzlich die Datierung 1531), zu beiden Datierungen s. bei A. Der durch diese Schenkung an Székely verursachte Textverlust in der Hs. in Neustift dürfte wohl dazu geführt haben, dass die entsprechenden Evangelientexte auf insges. neun Blättern handschriftlich ergänzt Cod. 196 219 wurden und bei der Neubindung in der 2. H. 18. Jh. miteingebunden wurden. Da in dem in Neustift erhal tenen Teil mit Bleistift zahlreiche Akzente zu finden sind, in jenem in Esztergom jedoch nicht, kann davon ausgegangen werden, dass die Hs. auch nach 1746 in Neustift noch verwendet wurde. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD Signaturen der ULB Tirol II 3 E 9 (Bleistift) und Nr. 196 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 196. Bl. 1r Stempel der ULB Tirol 19. Jh. Bei der Foliierung an der ULB Tirol wurden auch die in der 2. H. 18. Jh. neu ergänzten Blätter mitberück sichtigt (21–29, 36–42), 1872 wurden sie aber herausgeschnitten und ausgeschieden, vgl. Notiz Innsbruck, ULB Tirol, Alte Titelkopie: Handschriften: „als fremdartige durchaus werthlose Bestandtheile eines anderen Evangeliars und Epistolars, das mit dem vorliegenden prachtvoll ausgestatteten cumulirt war, im Beisein aller Beamter ausgeschieden und in die Makulatur gegeben und bei dieser Gelegenheit der alte Abgang der Feste Weihnachten bis Peter und Paul im II Theile des vorliegenden Werkes (Evangeliar) constatirt. 12/I/72. Leithe. Foregg. Hofer. Hörmann“. 1921 aufgrund des Friedensvertrages von St. Germain an den italieni schen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: 1 2 Wilhelm II 110. – Hermann 171–173, Nr. 182. – Atz 1012. – I. Genthon (Hrsg,), Esztergom műemlékei, T. 1 (Magyarország műemléki topográfiája 1). Budapest 1948, 350–354 (ohne Nennung der Sign.). – Neuhauser, Neustift 85. – Katalog ULB Tirol II 193, 209. – Peintner, Buchmalerei 35, 116–118 (ohne Nennung der Sign.). – Ch. Giner, Leben im Kloster, in: 850 Jahre Neustift. Katalog 85–90, hier 89. – Fiegl, Missale 19–20. – Roland, Renaissance 635, 640. – Baroffio 324, Nr. 16715. – E. Madas (in Verbindung mit K. Körmendy, J. Lauf, G. Sarbak), Katalog der mittelalterlichen Handschriften in Esztergom. Die Bestände der Kathedralbibliothek und anderer Sammlungen (Fragmenta et codices in bibliothecis Hungariae VII-B). Deutsche Fassung unter Mitwirkung von A. Vizkelety. Esztergom, Budapest (in Druck). (Ir–v) leer. (1r–20v) Epistolar. (1r) In galli cantu (Is 9,1 und 5–6). (1v) nur Rahmung und Linierung. (2r) In galli cantu (Tit 2,11–15). – In aurora (Is 61,1–3 und 62,11–12). (2v) (Tit 3,4–7). – (3r) Ad publicam missam (Is 52,6–10). (3v) (Hbr 1,1–12). – (4r) In nativitate sancti Stephani protho martiri (Act 6,8–10 und 7,54–60). – (5r) Iohannis ewangeliste (Eccl. 15,1–6). – (5v) In circum cisione domini (Gal 3,23–4,2). – (6r) Epiphanie domini (Is 60,1–6). – (6v) Agnetis virginis. (Eccl. 51,1–12). – (7r) Purificacionis. (Mal 3,1–4). – (7v) Ingenuini et Albuini (Eccl. 44,10– 15). – (8r) Annunciacionis (Is 7,10–15). – (8v) In consecracione palmarum (Ex 15,27–16,10). – (9v) In cena domini (1 Cor 11,20–32). – (10v) In festo pasce (1 Cor 5,7–8). – (10v) Feria IIa (Act 10,37–43). – (11r) Feria IIIa (Act 13,16–33). – (12r) Ascensionis (Act. 1,1–11). – (12v) In die Penthecostes (Act 2,1–11). – (13v) Feria IIa (Act 10,42–48). – (14r) Feria IIIa (Act 8,14– 17). – De sancta trinitate (Rm 11,33–36). – (14v) Corporis Christi. (1 Cor 11,23–29). – (15r) Iohannis Waptiste (Is 49,1–7). – (15v) Petri et Pauli (Act 12,1–11). – (16v) Visitacionis Marie (Ct 2,8–14). – (17r) Assumpcionis Marie (Sir 24,11–20). – (17v) Augustini episcopi (Hbr 5,1– 6). – (18r) Nativitatis Marie (Prv 8,22–35). – (19r) Michaelis (Apc 12,7–12). – (19v) Dedicacionis eclesie (Apc 21,2–5). – (20r) Omnium sanctorum (Apc 7,2–12). (30[21]r–35[26]r) Evangelistar, unvollständig. Der Beginn des Evangelistars [Bl. 1r–14v] befindet sich heute in Esztergom, Kathedralbibl., Ms. II.2 (s. bei G; ausführlichere Beschreibung mit Abb. in Genthon, Esztergom 350–354): [1r] In galli cantu. – [1v] In primo mane. – [2r] In die sancto. – [2v] In die s. Stephani. – [3r] Iohannis evangeliste. – [3v] In octava [nativitas]. – [3v] In die sancto epyphania. – [4v] In festo s. Agnetis. – [5r] Purificacio. – [6r] De patronis. – [6v] In annunciacione s. Marie. – [7v] In consecracione palmarum. – [8r] In cena domini. – [9r] In die sancto pasche. – [9v] Feria secunda. – [10v] Feria tercia. Expl. mut.: … nolite timere. Contur[ (1 Bl. fehlt, Textverlust) [11r] In ascensione dni. Inc. mut.: ] habebunt. Et dominus quidem Ihesus postquam locutus est eis assumptus … – [11r] In die penthecostes. – [11v] Feria IIa. – [12r] Feria IIIa. – [12v] De sancta trinitate. Expl. mut. … prohibebit de me et vos [ (1 Bl. fehlt, Textverlust) 220 Cod. 200 [13r] Vigilia Iohannis Baptistae. Inc. mut.: ] factum est autem cum sacerdotio fungeretur … – [13v] Evangelium Liber generationis. Require nativitatis Marie. – De corpore Christi. – Iohannis Baptiste. – [14v] Petri et Pauli. Expl. mut.: … cumque ligaveris super terram erit ligatum et in [ (30[21]r) Petrus et Paulus. Inc. mut. (Forts. von Esztergom, Kathedralbibl., Ms. II.2, Bl. 14v): ] celis et quidcumque solveris super terram erit solutum et in celis (Mt 16,19). – (30[21]r) De visitacione sancte Marie (Lc 1,39–56). – (30[21]v) In assumpcione s. Marie (Lc 10,38–42). – (31[22]r) In die Augustini (Lc 19,12–26). – (32[23]r) In nativitate s. Marie (Mt 1,1–16). – (33[24]r) In festo Michahelis (Mt 18,1–10). – (34[25]r) In dedicacione ecclesie (Lc 19,1–10). (34[25]v) In festo omnium sanctorum (Mt 5,1–12). (35[26]v) nur Rahmung und Linierung. U. S. Cod. 200 ROBERTUS HOLCOT Pap. I, 390, I Bl. 310 × 215. Wiener Raum (?), um 1415/20. B: Pap. stark fleckig, vereinzelt kleinere Einrisse am unteren Blattrand. Bl. 1 unten ergänzt sowie rechte Spalte kleines Loch, Bl. 158 eingerissen. Bl. 264–267, 276–277 sowie 287 und 290 etwas kleiner als restlicher Buchblock. Hinweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Iden tische Wasserzeichen: u. a. Wien, Schottenstift, Archiv, Hs. 101 (dat. 1416) und Archiv, Hs. 130 (dat. 1418). Lagen: (I–1)I (Vorsatzbl. 18. Jh.) + (VI–2)10 + 7.VI94 + V104 + 14.VI271(272) + (VI–1)282(283) + 8.VI378(379) + (VI– 1)389(390) + (I–1)I* (Nachsatzbl. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 9 und 10 (Textverlust), zu Bl. 281 (ohne Textverlust, Falz erhalten), zu Bl. 379 (Textverlust) herausgeschnitten. Bl. 9 am oberen Blattrand auf Bl. 8 aufgeklebt, Bl. 10 mit Papierstreifen mit Bl. 9 sowie 11 verbunden. Am Lagenende Kustoden in Rot ausgeschrieben, Lage 1–7 zusätzlich in Schwarz als arabische Ziffer. Ab Lage 8 Reklamanten, tw. beschnitten. In den Lagenmitten unbeschriebene Perga mentfälze. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 182 doppelt angeführt. S: Schriftraum 215/230 × 135/150. Zwei Spalten, tw. von feinen Tintenlinien, tw. von Blindlinien gerahmt, auf dem Verso vielfach ohne Rahmung, zu 38–45 Zeilen. Zirkelstiche. Bastarda von einer Hand mit unterschied lichem Duktus. Randbemerkungen und Korrekturen von späterer Hand. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Kustoden, Überschriften sowie Angabe der jeweiligen Lectio am oberen Blattrand. Bl. 1ra rote Zeigehand. Zwei- und dreizeilige rote Lombarden, tw. mit Punktverdickungen, tw. mit einfachen Ausläufern. Bl. 2va und 4va drei- bzw. zweizeilige rote Lom barde mit einfachem Fleuronnée in Schwarz (Bl. 4va: zwei gegenständige Halbpalmetten im Binnenfeld), Bl. 2va Fratze als Besatz. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. etwas abgerieben, bes. an den Kanten. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, tw. beschädigt und mit Wurmlöchern, drei Doppelbünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck ROBERTI HOLTGOT POSTILLA SUPER LIBRO SAPIENTIÆ in Goldrahmung, im dritten Signaturschild (s. bei G). Kapital oben freiliegend und beschädigt. Am unteren Schnitt Aufschrift Holdgot. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Bl. 377rb Besitzvermerk 15. Jh. Iste liber est monasterii sancte Marie in Novacella extra civitatem Brixinen sem sita. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`2 […] (radiert), II 3 E 12 (Bleistift) und Nr. 200 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 200. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 7416, 7416,3, 7416,12. – Kaeppeli Nr. 3497. – Katalog ULB Tirol II 201, 209. Cod. 204 221 (Ir–v) leer. (1ra–389vb) Robertus Holcot: Postilla super librum Sapientiae, unvollständig (Druck: GW 12884– 12890. – Stegmüller RB Nr. 7416. Kaeppeli Nr. 3497). (1ra) Prolog. Inc. mut.: ] usitata hec enim katholicos et in scismaticos et fideles in hereticos et doctores in adulatores convertit sicut recenter nobis occurrit et nuper exprimato vidistis. Hec enim facultas est illa ancilla Agar de qua scribitur… – Expl.: … quos precioso sanguine redemisti quod nobis ille con cedat qui cum patre vivit et regnat etc. (2va) Lectiones (insges. 212). Inc.: Diligite iusticiam qui iudicatis terram. Circa istum librum qui liber Sapiencie nuncupatur sunt in principio tria notanda … – Expl.: … Sapiencia deducet ad regnum perpe tuum. Ad quod regnum nos perducat ille qui cum patre et filio et spiritu sancto vivit et regnat in secula seculorum. Amen. Gloria tibi sit trinitas etc. (377rb) Schlussschrift (rot): Explicit postilla reverendi doctoris et magistri Roberti Holtgot ordinis fratrum Predicatorum super libros Sapiencie. Darunter: Besitzvermerk (s. bei G). (377v) leer. (378ra–389vb) Zwei Register. (378ra) Register der einzelnen Lectiones in alphabetischer Reihenfolge. Tit.: Incipiunt tituli questionum in exposicionem libri Sapiencie secundum ordinem alphabeti. – Inc.: Accepcio personarum utrum sit peccatum leccione 78 b … – Expl.: … Ymagines utrum sint adorande et quid in eis adoretur leccione 77 b. (379ra) Sachregister (unvollständig, A–H) (vgl. Register am Beginn des Drucks). Einführung: Ad evidenciam tabule sequentis que est super exposicionem libri Sapiencie. Sciendum quod in ea breviter continentur materie (am Blattrand ergänzt: naturales et) curiose questiones non tamen disputabiles … qui est superadditus propter prompciorem modum ad inveniendum materiam – Inc.: Aaron virga quomodo floruit leccione 93 b … – Expl. mut.: … Hospes preteriens est mundana felicitas [ (Ir–v) leer. U. S. Cod. 204 VETUS TESTAMENTUM Pap. 307 Bl. 310 × 200. Tirol (?), um 1470. B: Pap. am Beginn und am Ende des Buchblocks bes. am Rand, in der Mitte des Buchblocks im Bund ver schmutzt. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Iden tische Wasserzeichen; u. a. Innsbruck, ULB Tirol, Cod. 60 (um 1470/71); Neustift, Stiftsbibl., Cod. 174; Brixen, Priesterseminarbibl., Cod. A.2 (dat. 1472). Lagen: 25.VI301(300) + (V–3)308(307). Gegenbl. zu 304 und 306 herausgerissen (ohne Textverlust), Gegenbl. zu Bl. 302 als Spiegel auf dem HD aufgeklebt. Lagenrekla manten erhalten (außer bei Lage 24 und 25), tw. rot umrandet, ab Lage 10 beschnitten. In den Lagenmitten beschriebene und rubrizierte sowie illustrierte Pergamentfälze, stark verschmutzt. Vereinzelt Reste einer Zählung der Blätter der ersten Lagenhälfte erhalten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 260 ausgelassen. S: Schriftraum 210/220 × 140/145. Zwei Spalten, von Blindlinien gerahmt, zu 43–44 Zeilen auf Blindlinierung. Bastarda von zwei Händen: 1) Bl. 1ra–274ra, Z. 10; 2) Bl. 274ra, Z. 11–Bl. 306ra. A: Rote Auszeichnungsstriche, Unterstreichungen, Durchstreichungen, Rahmungen sowie Über- und Schluss schriften. Bl. 1ra–274ra Buchstaben in der obersten Zeile tw. gedehnt. An den Kapitelanfängen jeweils zwei- bis siebenzeilige rote Lombarden, meist mit Punktverdickungen, tw. mit einfachen Aussparungen, vereinzelt mit verzierten Ausläufern. Am Beginn der einzelnen Bücher fünf- bis elfzeilige rote Initialen mit 222 Cod. 204 Aussparungen in Form von Blättern (Halbpalmetten), ondulierenden Ranken und Kopfstempeln, Bl. 73va und 135va Herzblätter und/oder Blüten als Besatz. Bl. 119va und 120rb einfaches Fleuronnée in Schwarz. Repräsentanten. E: Gotischer Einband: dunkelbraunes Rindsleder über Holz mit Blinddruck. Tirol (?), Ende 15. Jh. Leder stark abgerieben, Holz z. T. freiliegend. VD und HD gleich: zwei durch je dreifache Streicheisenlinien begrenzte Rahmen, jeweils gefüllt mit Einzelstempeln. Rahmen 1: Einzelstempel Drache (Vogel?) in Raute (Nr. 1), in den Ecken Einzelstempel fünfblättrige Blüte frei (Nr. 2); Rahmen 2: vertikal abwechselnd Schrift band „Maria“ (Nr. 3) und Einzelstempel Einhorn in Raute (Nr. 4), in den Ecken Einzelstempel fünfblättrige Blüte frei (Nr. 2), unten und oben zudem je ein Einzelstempel Einhorn in Raute (Nr. 4). Im Mittelfeld oben und unten je ein Einzelstempel doppelköpfiger Adler in Raute (Nr. 5), in der Mitte Spuren eines rauten förmigen Beschlags. Ebenso in den Ecken Spuren von (spitz zulaufenden) Beschlägen. Zwei Leder-MetallSchließen, die obere schadhaft, die untere vollständig erhalten. Schließe mit Schriftband (wohl „Maria“). Kanten gerade. Rücken: drei Doppelbünde, Leder oben eingerissen, unten beschädigt. In den Feldern zw. den Bünden jeweils in den Ecken Einzelstempel heraldische Lilie in Raute (Nr. 6), in der Mitte Schrift band (Nr. 3). Im ersten Feld aufgeklebtes Papierschild M: S:, im zweiten Papiertitelschild SS Biblia sowie Signaturschild (s. bei G), im fünften (= letzten) Papierschild (leer). Kapitale mit rotem und blauem bzw. naturfarbenem Zwirn umstochen. Spiegel Pap. 15. Jh. Am Spiegel des VD Signaturen sowie Bleistiftnotiz (Vide Grundzettel), Spiegel des HD leer. G: Bl. 1r Besitzvermerk 18. Jh. Ad Collegium Neocell. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD Signaturen der ULB Tirol II 3 F 1 (Bleistift) und Nr. 204 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 204. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den ita lienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol III 25, 335. – Peintner, Stiftsbibliothek 121. – Baroffio 324, Nr. 16716. (1ra–306ra) Vetus Testamentum, unvollständig. (1ra–253vb) Libri historici: Pentateuchus, Iosua, Reges, Iudices, Ruth, Iudith, Esther. (1ra) ‹Hieronymus›: Epistola 53 (52) ad Paulinum presbyterum (BS I 3–37. Préfaces 1–7. – BHM Nr. 53. Stegmüller RB Nr. 284. CPL Nr. 620). (5ra) ‹Hieronymus›: Praefatio in Pentateuchum (BS I 63–69. Préfaces 7–8. – Stegmüller RB Nr. 285). – (5vb) Genesis. – (33ra) Exodus. – (56va) Leviticus. – (73va) Numeri. – (97vb) Deuteronomium. (119va) ‹Hieronymus›: Prologus in librum Iosue (BS IV 3–8. Préfaces 22–23. – Stegmüller RB Nr. 311). (120rb) Iosue. – (135va) 1. Regum. – (157vb) 2. Regum. – (170rb) 3. Regum. – (197rb) 4. Regum. – (217rb) Iudices. – (234rb) Ruth. (236vb) ‹Hieronymus›: Prologus in librum Iudith (BS VIII 213–214. Préfaces 35, Nr. 2. – Stegmüller RB Nr. 335). – (236vb) Iudith. (245va) ‹Hieronymus›: Prologus in librum Esther (BS IX 3–4. Préfaces 36, Nr. 3 und 4. – Stegmüller RB Nr. 341 und 343). – (245vb) Esther. (254ra–289vb) Libri Maccabaeorum. (254ra) 1. Maccabaeorum. – (275v) leer. – (276ra) 2. Maccabaeorum. (290ra–306ra) Iob. (306ra) Schlussschrift: Et sic est finis. Tu autem domine miserere nobis. (306rb–308v) leer. P. A. Cod. 217 223 Cod. 217 DECIMUS IUNIUS IUVENALIS. AULUS PERSIUS FLACCUS Perg. I, 56, I Bl. 310 × 200. Bologna, 1. H. 15. Jh. B: Perg. fleckig, insbes. am Blattrand Wasserflecken, löchrig, vielfach geflickt. Haar- und Fleischseite deutlich unterscheidbar. Perg. palimpsestiert (s. bei Inhalt). Auf Bl. 22v am oberen Blattrand je ein Farbstreifen in Grün (darin neuzeitlicher Eintrag uber?), Blau und Rosa (Pinselproben?). Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + III10(6) + II20(10) + 2.III70(22) + IV50(30) + II60(34) + Vunfoliiert(44) + II67(48) + IV80(56) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Zahlreiche Bl. fehlen (Textverlust), s. u. zeitgenössische Foliierung. Am oberen Blattrand zeitgenössische Foliierung. Heutige Bindung fehlerhaft, vgl. zeitgenössische Foliierung und Lagenreklamanten: 1–3, 8–10, 13, 11, 20, 18, 21–23, 28–30, 61–63, 68–70, 41–44, 47–50, 51–52, 59–60, zehn unfoliierte Bl., 64–67, 71–80. Am unteren Blattrand Foliierung 21. Jh. (hier berücksichtigt). Bl. 35 (Textbeginn der Satiren des Persius) neu zeitliches gelbes Papiersignakel der ULB Tirol. S: Schriftraum 220/225 × 125. Von feinen (am linken Textrand jeweils zweifachen) Tintenlinien begrenzt, zu einheitlich 25 Zeilen auf Tintenlinierung. Humanistische Minuskel. Interlinear- und Marginalglossen, bis weilen radiert, von drei zeitgenössischen Händen in Humanistischer Minuskel, tw. mit kursivem Einschlag. Vereinzelt Ergänzungen am Blattrand von späterer Hand. Bleistiftnotizen 19. Jh., insbes. die Anordnung der Blätter betreffend. A: Rubrizierung in hell- bis dunkelrosafarbener Tinte: Auszeichnungsstriche am Versbeginn (bis Bl. 2r), Über schriften, am oberen Blattrand und randseitig Nummerierungen (römische Ziffern) der Bücher und Satiren (auch in schwarzer Tinte), Randmarkierungen (z. B. Bl. 2v in der Art von vereinfachtem Perlenbesatz), Un terstreichungen, wenige Zeigehände. Repräsentanten. Zu Beginn der einzelnen Satiren ein- und zweizeilige rote und blaue Lombarden mit Fleuronnée, bei roten Lombarden (stark verblasstes) rosafarbenes, bei blauen Lombarden rotes Fleuronnée (Bl. 17r einzeilige rosafarbene Lombarde mit zierstabartigem Mittelstrich). In den Binnenfeldern und als Besatz charakteris tische saitenartige Parallelstriche, verziert mit kleinen Perlen; randseitig aus den durch Schräghaken in gegeneinander versetzte Abschnitte unterteilten Parallelstrichbündeln einzelne Striche zu schwungvollen Fadenausläufern verlängert, charakteristische große, nach unten weisende Schlingen mit (hakenartigem) Binnenmotiv. Erhaltener Deckfarbenschmuck: vier rankenlose Ornamentalinitialen, sechs- und vierzeilig am Buchbeginn Bl. 1r und 35r, zweizeilig am Anfang von zwei Satiren, Bl. 9v und 27r. Blattvergoldete Außengründe (Bl. 1r rosafarbener Bolus sichtbar) mit schwarzen Rahmenlinien, fallweise mit feinen Eckspitzen; rosafarbene Buchstabenkörper (Bl. 1r mit grünen „Anschnitten“ und roter Manschet te, Bl. 35r mit Serifenblatt); in den Binnenfeldern auf blauem Grund diverse Blattformationen in Grau, Grün und kräftigem Rot; feiner weißer Lineardekor; Buchstabenkörper und Blätter mit (angedeutetem) „Erb senschotenmotiv“; neben den Initialen auf dem Seitenrand freie Goldpunkte (mit schwarzer Konturlinie, Strahlen und kleinen Kreisen). Das Kolorit (u. a. das Blau der Binnengründe), die feinen weißen Dekorlinien und das „Erbsenschotenmo tiv“ verweisen auf die italienische Buchmalerei 1. H. 15. Jh.; das Fleuronnée ebenfalls italienisch. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. stark abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck IUVEN. ET PERSII WITR. (!) SATY. in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Signa turen und Bleistiftnotizen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Aufhe bung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`5 C 4 (Tinte), II 3 F 14 (Bleistift) und Nr. 217/(1) (Tinte, Bleistift) sowie Bleistiftnotizen (Vide Grundzettel) (die Textblätter des Juvenal u. Persius sind verwechselt). Am Rücken gelbes Signaturschild der ULB Tirol 217. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller VI 122. – Wilhelm I 136. – Hermann 173, Nr. 183. – A. Zingerle, Ueber eine Innsbrucker Juvenal handschrift mit Scholien (Kleine philologische Abhandlungen 4). Innsbruck 1887, 1–12. – A. Zingerle, 224 Cod. 217 Übersicht über philologische Handschriften aus Tirolischen Bibliotheken. Commentationes Aenipontanae 4 (1909) 1–4, hier 3, Nr. 8. – Neuhauser, Neustift 84. – Katalog ULB Tirol III 64, 335. – Peintner, Stiftsbiblio thek 122. – M. Peintner, Das Skriptorium von Neustift, in: 850 Jahre Neustift. Katalog 91–93, hier 93 (ohne Nennung der Sign.). 1 (Ir–v) leer. (1r–34v) Decimus Iunius Iuvenalis: Saturae, unvollständig (Ed. I. Willis. Stutgardiae et al. 1997). Aufgrund fehlender Blätter lückenhafter Text mit reichlichen Interlinear- und Randglossen von drei unterschiedlichen Händen. Den einzelnen Satiren sind die entsprechenden Argumenta von Guarinus Veronensis vorangestellt (Ed.: D. Manzoli, Nuovi carmi di Guarino Veronese. Verona 2000). Am oberen Blattrand: Liber primus satyra prima. Tit.: Iunii Iuvenalis Aquinatis poetarum satyricorum facetissimi et gravissimi liber Satyrarum suarum primus incipit. Aufgrund der fehlerhaften Bindung ergibt sich folgende Reihenfolge: (1r) Sat. 1, V. 1–146. Tit.: Materiam et causas satyrarum hac inspice prima. – (4v) Sat. 3, V. 3–152. – (7r) Sat. 3, V. 253–302. – (8r) Sat. 3, V. 153–202. – (9r) Sat. 5, V. 134–173. – (9v) Sat. 6, V. 1–15. Tit.: Hac (!) satyra incestas mulieres monstrat, ut omnem uxorem Ursidius devitet pectore toto. – (10r) Sat. 5, V. 26–75. – (11r) Sat. 6, V. 16–169. – (14r) Sat. 6, V. 371–521. – (17r) Sat. 12, V. 126–130. – (17r) Sat. 13, V. 1–146. Tit.: Mens cruciat pravos. Sapiens fert damna modeste. – (20r) Sat. 14, V. 97–247. – (23r) Sat. 8, V. 119–275. – (26r) Sat. 9, V. 1–45. Tit.: Con cubitus diri et sodomitica probra notantur. – (27r) Sat. 10, V. 1–294. Tit.: Arguit hec hominum curas et inania vota. Von anderer Hand: Liber quartus. – (33r) Sat. 12, V. 25–125. Zur Kommentierung Iuvenals s. auch E. M. Sanford, Renaissance Commentaries on Juvenal. Transactions and Proceedings of the American Philological Association 79 (1948) 92–112. 2 (35r–44v) Aulus Persius Flaccus: Saturae, unvollständig (Ed. W. Kissel. Berolini 2007). Tit.: A. Persii Vultrani satyrarum libellus incipit feliciter. (35r) Prolog (hier als Satura 1 gezählt). Schlussschrift: Explicit prima. (35r) Sat. 1. Tit.: Incipit secunda de perversis exercitationibus poetarum. – (38r) Sat. 2. Tit.: Satira II de falsis votis. – (39v) Sat. 3. Tit.: Satyra III de inerta (!) iuvenum. – (41v) Sat. 4. Tit.: S[atura] IIII in ambitiosos cives. – (43r) Sat. 5, V. 1–99. Tit.: S[atura] V ad Corntum (!) poetam de illecbris (!) urbis. 3 (45r–53r) Forts. von Nr. 1: Decimus Iunius Iuvenalis: Saturae, unvollständig (s. o.). (45r) Sat. 13, V. 147–249. – (47r) Sat. 14, V. 1–96. Tit.: Imbuit exemplis natos mala vita paren tum. – (49r) Sat. 14, V. 248–331. – (50v) Sat. 16 (am Rand als XV gezählt), V. 1–60. Tit.: Militiae mores et commoda dinumerantur. – (51v) Sat. 15 (am Rand als XVI gezählt), V. 1–54. Tit.: Im manes ritus Aegyptius incola servat. – (53r) Sat. 15, V. 155–174. (53v–56v) leer. (Ir–v) leer. Gesamte Hs. palimpsestiert. Möglicherweise handelt es sich um ein Register für den Stationarius (wohl eines Notars) zur Dokumentation des Verleihs von Pecien; bes. auffallend ist die Verwendung der lat. Sprache. Aufgrund inhaltlicher, u. a. topographischer Hinweise im Umfeld der Universität Bologna entstanden. Giovanna Murano sei für ihre wertvollen Hinweise herzlich gedankt. G. K. / A. P. Cod. 252 225 Cod. 252 ORDINARIUM MISSAE Perg. 46 Bl. 340 × 245. Neustift (?), letztes Viertel 15. Jh. B: Dickes Perg. Lagen: (IV+1)9 + 4.IV41 + (III–1)46. Am Beginn fehlt eine Lage (vermutlich Quaternio). Bl. 1 eingehängtes Einzelbl., Gegenfalz zu Bl. 1 zw. erster und zweiter Lage. Gegenbl. zu Bl. 42 herausgeschnit ten (Textverlust). Am rechten unteren Blattrand Lagenfoliierung 1–8. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Bl. 1 Spuren eines Signakels. S: Schriftraum 225/255 × 165/175, von Tintenlinien gerahmt, Bl. 2r zu zehn, Bl. 2v–25v (Canon Missae) zu einheitlich neun Zeilen, Bl. 26r–46v zu einheitlich 14 Zeilen auf Tintenlinierung. Zirkelstiche. Textualis formata (Textura). A: Rote Auszeichnungsstriche; bei k und Q die Serifen etwas langgezogen, wodurch sie wie Unterstreichungen wirken; rote Benediktionskreuze in quadratischem Rähmchen; ein- und zweizeilige rote Lombarden. Reprä sentanten. Bl. 2r Te-igitur-Initiale (insgesamt dreizeilig) mit der Vorzeichnung eines zweizeiligen T, dessen Querbalken auf den mit einem Lilienknauf bekrönten Längsschaft aufgesteckt ist. Bl. 1v ganzseitiges Kanonbild (245 × 175): dreifigurige Kreuzigung in qualitätvoller Deckfarbenmalerei. In einem plastisch wirkenden grünen Profilrahmen (Breite 14 mm) mit quadratischen, einfach punzierten Gold applikationen an den Ecken und in der Mitte der Längsseiten, vor karminrotem, mit feinem Goldmuster (ge stricheltes Rautengitter mit eingeschriebenen Ästchen) überzogenem Grund Christus an einem hohen TauKreuz mit naturgetreu herausgearbeiteter Maserung, das auf einer verhältnismäßig kleinen, von Knochen umgebenen Felsformation steht. Der Körper des Gekreuzigten anatomisch weitgehend korrekt dargestellt (s. v. a. Armmuskeln, Brustkorb sowie Knie- und Wadenpartie). Die Inkarnate korrespondieren mit dem hel len Pergamentton des Folios (das Lendentuch Christi nur wenig heller), Modellierung und Schattierung mit Weißhöhungen und Grau- bzw. Rosatönen. Während Christus die Augen geschlossen hat, blicken Maria und Johannes, die auf hügeligem, in weichem Grün koloriertem Wiesenboden (mit tw. größeren Pflanzen) stehen und sich dem Kreuz zuwenden, zu Boden. Der Gekreuzigte und Johannes mit fein gewellten Haaren; alle drei Figuren mit Goldnimben (Blattgold auf rosafarbenem Bolus, leicht beschädigt, mit Punktlinien radial pun ziert), bei Christus zusätzlich mit blauem Kreuz; charakteristisch die relativ langen, aber rundlich endenden Nasen. Maria hat beide Hände vor die Brust erhoben; Johannes, dessen Hände noch zarter gestaltet sind als die der Gottesmutter, hält seine Rechte an die Wange. Gekonnt herausgearbeitete Draperie durch ausgewoge ne Knickfalten und Konturlinien, Schattierungen in der Lokalfarbe und wenige Schraffuren. Verschmutzun gen im unteren Bereich des Marienmantels, der Beine Christi und auf dem Basisfelsen. Keine Ranken rahmung, lediglich die goldenen Rahmenbeschläge jeweils mit zartem Floraldekor in Karminrosa besetzt: an den Längsseiten Dreiblätter und an den Ecken je zwei kurze, überkreuzte Ästchen mit erkennbaren Anschnitten. Die unfigürlichen Anteile des Kanonblatts dürften von einem anderen Illuminator geschaffen worden sein als die Figuren. Während sich für diese Hand keine Entsprechungen in der Neustifter Buchmalerei nachwei sen lassen, finden sich für den Ornamentalgrund der Kreuzigung und den Blättchendekor des Rahmens Vergleichsbeispiele im Epistolar des Cod. 196 (figürliche Initiale Bl. 5v dat. 1482): Bl. 14r der feine Gold ästchendekor auf dem Initialfeldgrund, Bl. 12v das angeschnittene, neben der Initiale in die Ranke ein gehängte Ästchen und Bl. 17r (links unten) die Gestaltung der rosafarbenen Rankenblättchen. S. R. E: Gotischer Einband: blau gefärbtes Wildleder über Holz mit Blindlinien. Neustift (?), Ende 15. Jh. Leder stark abgerieben, fleckig, Holz tw. freiliegend. Zwei Schichten: tw. freiliegendes Schweinsleder, darüber blau gefärbtes Wildleder mit Blindlinien. VD und HD gleich: Gliederung durch jeweils dreifache Streicheisenlinien: ein von solchen Linien begrenzter Rahmen, Mittelfeld horizontal, vertikal und diagonal unterteilt, zudem Raute eingeschrieben. Spuren von je fünf runden Beschlägen. Zwei Leder-Metall-Schlie ßen. Kanten gerade. Rücken: unten tw. freiliegend, Leder beschädigt, fünf Doppelbünde, jeweils durch ein fache Streicheisenlinien betont. Im ersten Feld aufgeklebtes Papierschild M: S:, im zweiten aufgeklebtes Papierschild, beschädigt Fragmenta Varia Miss[ale], im dritten und vierten Signaturschilder (s. bei G). Oberes Kapital mit naturfarbenem Zwirn umstochen, unteres Kapital fehlt. Spiegel Pap. 15. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. und alte Signaturen der ULB Tirol II`1 (korr. aus 2) A 10 (Bleistift) und 252 (Tinte) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken Reste des gelben 226 Cod. 252 ignaturschilds der ULB Tirol. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der S ULB Tirol übergeben. Bl. 2r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe übergeben. Bl. 2r Stempel der Soprintendenza Tridentina 20. Jh. Am Rücken Neustifter Signatur schild 20. Jh. 252. L: Wilhelm I 110. – Hermann 173, Nr. 184. – Katalog ULB Tirol III 204, 335. – Baroffio 324, Nr. 16717. Ordinarium missae. Enthält Canon Missae bis Schluss der Messe, Gebete nach der Messe, Präfationen, Votivmessen und Totenmessen. Beginn (Messtexte bis Canon missae) aufgrund des Verlustes einer Lage nicht mehr enthalten. Aufgrund des zeitgenössischen Einbandes ist anzunehmen, dass es sich nicht um ein heute unvollständiges Vollmissale handelt, sondern um ein Teilmissale (Ordinarium missae), dessen Beginn (s. bei B) verloren ist. Ohne Notation. (1r) leer. (1v) ganzseitiges Kanonbild (s. bei A). (2r–21v) Canon missae. (2r) Von Te igitur bis Doxologie. (12v) Pater noster. (13v) Libera nos. (14v) Pax dni. (15r) Agnus Dei. (15r–21v) Kommuniongebete. (21v) Gebet Post missam. Placeat tibi sancta trinitas hoc obsequium servitutis mee et presta ut hoc sacrificium … (22v) Schlussevangelium (nur Rubrik und Textbeginn) und Segen. (22v–24v) Orationes post missam (vgl. Jungmann II 571–572). Ablegung der Paramente mit Verweis auf Canticum trium puerorum und Textbeginn von Ps 150, Can ticum Simeonis (Lc 2,29–32) und Vermerk (rot) Per totum cum gloria Patri antiphona (schwarz) Trium puerorum ut supra, Kyrie, Pater noster, fünf Versikel, zwei Gebete: (24r) Deus qui tribus pueris miti gasti flammas ignium … und (24r) Acciones nostras quesumus domine aspirando preveni et adiuvando prosequere … sowie Vermerk (24v) (rot) Tunc flectis genibus dic (schwarz) Laus patri glorie laus eius filio laus quoque sancto spiritui sit per eum Amen. (24v) Gloria. (25v) Credo. (27r–32v) Praefationes und Communicantes. (27r) De nativitate domini. – (27v) De epyphanie (!) domini. – (28r) In Quadragesima. – (28v) In cena domini. – (29v) De resurreccione domini. – (30r) De ascensione domini. – (30v) In festo penthe costen. – (31v) De s. trinitate. – (32r) De sancta cruce. – (32v) De beata virgine. – (32v) De apostolis. – (33r) Praefacio cotidiana. (33v–42r) Missae votivae. (33v) In festo penthecosten. Mit Sequenz (AH 53, Nr. 70). – (38r) De sancta cruce. – (40r) Sabbato de commemoracione b. Marie. Mit Sequenz (AH 54, Nr. 218). (42r–46v) Missae pro defunctis, unvollständig. (42r) Officium pro defunctis. – (44v) Pro elemosinariis. – (45r) Pro famulis et famulabus. – (46r) Pro fidelibus defunctis. – (46v) Pro sacerdote. Unvollständig aufgrund des Verlustes des letzten Blattes der letzten Lage. Schließt mit Beginn der Secreta. Expl. mut.: Suscipe quesumus [ W. N. Cod. 253 227 Cod. 253 GODEFRIDUS DE TRANO. TANCREDUS BONONIENSIS. RAIMUNDUS DE PENNAFORTE Perg. 88 Bl. 380 × 265. Oberitalien (Bologna?), 1. H. 14. Jh. B: Drei Teile: I (1–82), II (83–84), III (85–88). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. E: Gotischer Einband: braunes Leder über Holz mit Blindlinien. Oberitalien (Bologna?), 15. Jh. Leder stark abgeschabt, tw. beschädigt, darunter liegendes Textil sichtbar. VD und HD gleich: aus dreifachen Streicheisenlinien gebildeter Rahmen. Mittelfeld durch ebensolche Linie diagonal unterteilt. Spuren eines runden Mittelbeschlags sowie von vier Eckbeschlägen (Viertelkreis). Spuren von vier Kurzriemen-LederMetall-Schließen (oben, unten, seitlich), Nägel erhalten. Am VD oben Titelaufschrift am Leder Summa Goffridi. Am HD Aufschrift E I […]. Kanten gerade. Rücken: vier Doppelbünde. Im ersten Feld Papierschild M S, im zweiten Ius Canonicum Glossarum, im dritten Signaturschild (s. bei G). Kapitale mit naturfarbenem Spagat umstochen. Spiegel des VD und HD mit Textil überzogen, am Spiegel des VD Exlibris (s. bei G). G: Die Hs. dürfte wohl zu jenen vier Hs. kirchenrechtlichen Inhalts zählen, die 1353 als Geschenk des Herrn Berthold, Hospitaler zu Brixen, in den Besitz des Stiftes gelangten (vgl. Neustift, Stiftsarchiv, WW 59. Mair hofer 364–365; Peintner, Stiftsbibliothek 118). Bl. 1r Besitzvermerk Ad Collegium Neocell. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r alte Signaturen der ULB Tirol II`1 A 11 (Bleistift) und 253/(I) (Tinte, Bleistift) sowie Bleistiftnotizen (Wird behandelt u. katalogisiert) und (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 253/(I). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Schulte GQ I 203, II 89. – Wilhelm I 136. – Wretschko–Sprung 17–18. – Dolezalek I o. S. – Furtenbach– Kalb 20. – Katalog ULB Tirol III 204, 335. I (1–82) B: Haar- und Fleischseite deutlich unterscheidbar, unregelmäßig beschnitten (mit Fehlstellen). Bl. 82 von klei nerem Format (380 × 145/150). Lagen: 10.IV80 + I82. Lagenreklamanten. S: Schriftraum 290 × 205. Zwei Spalten, von feinen, bisweilen kaum sichtbaren Bleistift- bzw. Blindlinien be grenzt, zu 59 Zeilen auf Bleistift- bzw. Blindlinierung. Zirkelstiche. Textualis. Am Blattrand Nachträge, Korrekturen und Bemerkungen von verschiedenen Händen. A: Rote Unter- und Durchstreichungen sowie Überschriften. Zeigehände. Abwechselnd blaue und rote Para graphzeichen sowie zwei- und dreizeilige Lombarden mit einfachem Fleuronnée und Fadenausläuferbün deln in der Gegenfarbe. Repräsentanten. Am Blattrand diverse Verweiszeichen. Am oberen Blattrand blau- rote Buchbezeichnung. Bl. 62v und 63r schwarze Federzeichnungen: Arbor consanguinitatis und Arbor affinitatis, jeweils mit Zierrahmen. (1r, am oberen Blattrand) Ave Maria gratia plena dominus tecum. 1 (1ra–82ra) Godefridus ‹de Trano›: Summa super rubricis Decretalium (Druck: u. a. GW 10949. – Stegmüller RS Nr. 245. Stegmüller RS [1937] Nr. 904). Tit. (18. Jh.): Summa Gaffridi. De Regulis Juris. (1ra) Prolog. Inc.: [G]losarum diversitas intelligentiam textus nec numquam obtenebrat … – Expl.: … aliorum tales alios esse volo intellectores meorum. (1ra) Text. Inc.: Post proemium in quo breviter edocemur … – Expl.: … et propterea malui repeti quam deesse. (62v–63r) Zwei Federzeichnungen: Arbor consanguinitatis und Arbor affinitatis. (82r) Schlussschrift: Explicit liber Deo gratia. Ave Maria gratia plena dominus tecum. Benedicta tu in mulieribus et benedictus fructus ventris tui. Gloria sit Christo per infinita secula seculorum Amen. (82v) leer. Lit.: Schulte GQ II 89–91. 228 Cod. 253 II (83–84) B: Zwei Einzelbl.: Lage: 183 + 184. Fragmente, einst möglicherweise als Spiegel aufgeklebt (Rostspuren, Klebe reste), Faltung und Bindung (quer zum heutigen Schriftraum) ersichtlich. Bl. 83 beschnitten (340 × 160), heute lose. Bl. 84 (355 × 210) beinahe lose, mit Papierstreifen oben und unten auf Falz aufgeklebt. Bl. 84ra bzw. vc beschnitten. S: Erhaltener Schriftraum 165 × 65. Drei Spalten, von Blindlinien gerahmt, zu 46 Zeilen auf Blindlinierung. Textualis. 2 2.1 (83ra und 84ra–c) Tancredus ‹Bononiensis›: Ordo iudicarius, unvollständig. (83ra) Tancredus ‹Bononiensis›: Ordo iudicarius, unvollständig: Pars 4, tit. 6, § 5–7 (Forts. von Bl. 84ra–c, ohne Textverlust) (Ed.: F. Bergmann, Pillii, Tancredi, Gratiae libri de iudiciorum ordine. Göttingen 1842, 312–314). Inc. mut.: ]ma. sed etsi § quocumque quomodo intra diem illam poterit probare lesionem … – Expl.: … ad rudium instructionem breviter dixisse sufficiant Amen. Schlussschrift: Explicit liber Tancredi. (83rb, quer zum Schriftraum) Interesse quinque sunt dampnum lucrum amissum. (83v) Federproben: Ista est summa domini Acrolini … 3 (83v) Urkundenvorlage (anonymisiert): Inc.: [V]enerabilibus in Christo patribus dominis Dei gratia archiepiscopis et episcopis sedis apostolice gratiam et communionem habentibus … – Expl.: … munitas dant etc. 2.2 (84ra–c) Tancredus ‹Bononiensis›: Ordo iudicarius, unvollständig: Pars 4, tit. 5, § 14–tit. 6, § 5 (Forts. von Bl. 84va–c, ohne Textverlust) (Ed. Bergmann 300–312) Inc. mut.: cor]rigebatur quod in ecclesia sua nolebat facere residentiam … – Expl. mut.: … ad restitutionem optinendam ut expresse dicit 1 illa ff ex qui cau. [ (84va–c) Pars 4, tit. 5, § 3–14 (Ed. Bergmann 288–300). Inc. mut.: ] sibi adiudicate ut extra de sentencia et re iu. c. Bertoldus in fi. Si autem personali … – Expl. mut.: … tenet eius appellacio ut extra de ap. pervenit de illo clerico qu[i Lit.: Schulte GQ I 203–204 und II 558; Kuttner 431–433. III (85–88) B: Perg. fleckig, Haar- und Fleischseite deutlich unterscheidbar, Bl. 86 Loch. Lage: II88. S: Schriftraum 165 × 65. Drei Spalten, von Bleistiftlinien gerahmt, zu 78 bzw. 80 Zeilen auf Bleistiftlinierung. Zirkelstiche. Textualis. A: Bl. 85rc–87rc rote Überschriften. Repräsentanten am Blattrand. 4 (85ra–88vc) ‹Raimundus de Pennaforte›: Summa de matrimonio, unvollständig (Ed.: X. Ochoa, A. Diez [Universa Bibliotheca iuris I C]. Roma 1978, 901–998. – Kaeppeli Nr. 3408. Bloomfield Nr. 4943 und Bloomfield, Suppl. Nr. 4943). (85ra–88rc) Forts. des Textes von Bl. 88rc–vc, ohne Textverlust (Tit. 2,7–25 und Conclusio). Inc. mut.: ] indicas Hermanum VII q. I Quamvis triste et C. de codicillis nec codicillos quod vero supra dictum est … – Expl.: … sed benigno corrigat et emmendet (= Schluss). Schlussschrift: Explicit summa de matrimonio. (88rc–vc) Textbeginn (Prooemium und Tit. 1–2,7): Tit.: Incipit summa de matrimoniis. – Inc.: Quoniam frequenter in foro penitentiali dubitationes circa matrimonium … – Expl. mut.: … omnia que alii possunt ut est ar. III q. VIIII Indicas Her manum. Require superius Indicas Hermanum per […] cartulas et ibi invenies [ Darunter Schlussschrift (fälschlich): Explicit summa matrimonialis. C. S. Cod. 282 229 Cod. 282 BERNARDUS CLARAEVALLENSIS Perg. I, 25, I Bl. 240 × 165. Entstehungsort unbestimmt, 2. H. 12. Jh. B: Perg. löchrig, Nahtspuren. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 2.IV16 + (1+IV)25 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Am oberen Blattrand mittig zeitgenössische Foliierung in arabischen Ziffern 89–113. Vor erster Lage Textverlust. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol 1–25 (hier berücksichtigt). Bl. 15r neuzeitliches oranges Papiersignakel der ULB Tirol. S: Schriftraum 170 × 105, von feinen, tw. kaum sichtbaren, seitlich zweifachen Bleistiftlinien gerahmt, zu 34 Zeilen auf Bleistiftlinierung. Zirkelstiche. Karolingisch-gotische Mischschrift. Bl. 9v und 20v Randglossen von späterer Hand, Bl. 21r Federprobe. A: Rote Auszeichnungsstriche (nur Bl. 1r), Über- und Schlussschriften. Vereinzelt Randzeichen. Zwei- bis fünfzeilige rote bzw. grüne Lombarden, tw. mit einfachem Dekor in der Gegenfarbe. Bl. 12r rote Satzma juskel in Lombardenform. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. VD und HD beschädigt. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, drei einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S: und Signaturschild (s. bei G), im zweiten und dritten Golddruck BERNARD. CLAREVALL. in Goldrahmung, im vierten Leder abgeschabt, urspr. wohl Schild aufgeklebt. Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftnotiz (s. bei G). Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`1 C 1 2. Ser. (Bleistift) und 282/1 (Tinte, Blei stift) sowie Bleistiftnotiz (Vide Grundzettel). Am Rücken gelbes Signaturschild der ULB Tirol 282. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 135. – Leclercq–Rochais III 7, 159. – BGOC, Bernard Nr. 4193. – E. Manning, H. Rochais, Bi bliographie Générale de l’Ordre Cistercien. Saint Bernard. Documentation Cistercienne 21, fasc. hors série 10–11–12. Rochefort 1982, Nr. 4193. – Katalog ULB Tirol III 279, 335. 1 (Ir–v) leer. (1r) Bernardus Claraevallensis: De gratia et libero arbitrio, unvollständig: cap. 14,51 (PL 182, 1029A–1030A. Ed. Leclercq–Rochais III 203. – BGOC, Bernard Nr. 4193). (1r) Inc. mut.: ] contempnens suam velit statuere iusticie domini non sit subiectus … Schlussschrift: Explicit liber domini Bernhardi Clarevallensis abbatis de gratia et libero a rbitrio. 2 (1r–15r) Bernardus Claraevallensis: De gradibus humilitatis et superbiae (PL 182, 939–972C. Ed. Leclercq–Rochais III 15–59. – BGOC, Bernard Nr. 4193) 3 (15r–25v) Bernardus Claraevallensis: Epistola 42 (ad Henricum Senonensem archiepiscopum: De moribus et officio episcoporum) (PL 182, 809A–834A. Ed. Leclercq–Rochais VII 100–131. – BGOC, Bernard Nr. 4193). Tit.: Epistola domni (!) Bernhardi abbatis Clarevalensis ad H. Senonensem archiepiscopum. (Ir–v) leer. P. K. 230 Cod. 293 Cod. 293 VITA BEATI HARTMANNI EPISCOPI (BRIXINENSIS) Perg. 23, II Bl. 260 × 170. Neustift, 12./13. Jh. B: Dickes, fleckiges, tw. löchriges, an den Rändern unregelmäßig beschnittenes Perg., vereinzelt Nahtspuren. Bl. 22 untere Blatthälfte nach 15 cm weggeschnitten, Bl. 23 an dieser Stelle eingeschnitten. Lagen: IV8 + (IV–1+1)16 + (IV–1)23 + IIII* (Nachsatzbl. Pap.). Gegenbl. zu Bl. 11 herausgeschnitten und durch eingefügtes Blatt ersetzt (gleicher Schreiber), jetzt tw. lose (= Bl. 14). Gegenbl. zu Bl. 18 herausgeschnitten (ohne Text verlust). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Bl. 1r–21v Schriftraum 200/210 × 130, von kaum wahrnehmbaren Tintenlinien gerahmt, zu meist 27 Zeilen auf Blindlinierung. Zirkelstiche. Karolingisch-gotische Mischschrift 12. Jh. Bl. 15v Z. 19–27 möglicher weise von anderer Hand, vgl. Sparber 1940, 10. Überschrift Bl. 1r in Textualis 13. Jh., Ergänzungen am Rand von Hand des Schreibers, Randnotizen in Textualis 13. Jh. und Notula Mitte 15. Jh. (gleiche Hand wie die Zusätze in Innsbruck, ULB Tirol, Cod. 960, dort Schreiber C, vermutlich der Neustifter Stiftschronist Johannes Librarius [Kellner], s. W. Neuhauser, Beschreibung und Geschichte der Handschrift, in: E. Thurn her, W. Neuhauser (Hrsg.), Die Neustifter-Innsbrucker Spielhandschrift von 1391, Cod. 960 der Universi tätsbibliothek Innsbruck [Litterae 40]. Göppingen 1975, 11). Bl. 21v Ergänzung am Schluss in Notula 15. Jh. Bl. 22 und 23 Schriftraum und Zeilenzahl wechselnd. Textualis 13. Jh. und Bastarda 15. Jh. A: Bl. 1r–21v rote Auszeichnungsstriche, Unterstreichungen, Satzmajuskeln in Lombardenform, zwei- bzw. dreizeilige Lombarden. Bl. 1r fünfzeilige rot-blaue Initiale F mit blauer Füllung, weißen Aussparungen und kurzen blattförmigen rot-blauen Ausläufern. Bl. 22 und 23 rote Überschriften, Unterstreichungen, Auszeich nungsstriche, Paragraphzeichen, Bl. 23r zweizeilige Lombarde E. E: Makulatureinband: Perg. über Pappe, Neustift, 15. (?) Jh. VD, Rücken und HD von durchgehendem Perg.-Doppelbl. überzogen. Fragment eines Antiphonale, erhalten Ausschnitte zum Ostersonntag: sieben rote Vierliniensysteme mit gotischer Choralnotation und Textzeilen, Textualis formata (Textura) 14. Jh. Rote Überschrift, Initiale A und zwei Cadellen mit einfachem Fleu ronnée. Am HD Stempel der Soprintendenza Tridentina. Spiegel Pap. 15 Jh. Am Spiegel des VD Exlibris, Signaturen und Bleistiftvermerk (s. bei G), Spiegel des HD leer. G: Bl. 23v Besitzvermerk 15. Jh. Iste liber est monasterii s. Marie virginis in Novacella. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neu stift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`6 B (?) […] (Tinte, radiert), II1 C 12 S. 2 (Bleistift) und 293 (Tinte) sowie Bleistiftvermerk (Vide Grundzettel). 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgetreten und von diesem 1929 dem Klos ter Neustift als Dauerleihgabe überlassen. Am HD Stempel der Soprintendenza Tridentina 20. Jh. L: H. Pez, Scriptores rerum Austriacarum I. Lipsiae 1721, 495–518 (= erste Ed., ohne Nennung der Sign.). – Ph. N. Puell, Heiligmäßiger Lebenswandel des seligen Hartmanni. Brixen 1768, 180 Anm. 2 (ohne Nen nung der Sign.). – H. R. v. Zeissberg, Zur Kritik der Vita B. Hartmanni episcopi Brixinensis. Archiv für österreichische Geschichte 56 (1878) 454 (mit Anführung der alten Sign. der ULB Tirol II`1 C 12 S. 2). – Wilhelm I 135. – A. Sparber, Der selige Hartmann. Brixen 1911, 87 (ohne Nennung der Sign.). – Sparber 225–226. – A. Sparber, Vita beati Hartmanni episcopi Brixinensis. Einleitung und Ausgabe. Brixen (als Manuskript gedruckt) 1939. – Vita beati Hartmanni episcopi Brixinensis. Eingeleitet und ed. von A. Sparber (Schlern-Schriften 46). Innsbruck 1940. – A. Sparber, Leben und Wirken des seligen Hartmann, Bischof von Brixen (1140–1164). Wien 1957. – A. Lhotsky, Quellenkunde zur mittelalterlichen Geschichte Österreichs (MIÖG Erg.-Bd. 19). Graz, Köln 1963, 207. – Neuhauser, Neustift 81. – Steer 36. – Katalog ULB Tirol III 309, 336. – G. Kompatscher, Überblick, in: M. Korenjak, F. Schaffenrath, L. Šubarić, K. Töchterle (Hrsg.), Tyrolis Latina. Geschichte der lateinischen Literatur in Tirol I: Von den Anfängen bis zur Gründung der Uni versität Innsbruck. Wien, Köln, Weimar 2012, 31–52, hier 37. – Baroffio 324, Nr. 16718. – S. C. Schmölz, Die liturgische Verehrung des seligen Hartmann von Brixen, Dipl.-Arb. St. Pölten 2012, 88 („Hs. A“, ohne Nennung der Sign.). 1 (1r–21v) Vita beati Hartmanni episcopi (Brixinensis), Rec. I (Ed. Pez 495–518 [= erste Ed.]. Sparber 1940, 37–62. Schmölz 95–117). Tit.: Prohemium in vitam beati Hartmanni episcopi. – Inc.: Fecit Moyses labrum eneum de spe culis mulierum quem plenum aqua posuit … – Expl.: … cum ceteris sanctis ac fidelibus Christi Cod. 293 231 dignam reverentiam recipiat. Bl. 21v nach recipiat am Schluss Ergänzung (15. Jh.) cui sit gloria et honor per infinita secula seculorum Amen. Cod. 293 in der Überlieferung Hs. A der Rec. I. Zur Überlieferung s. Sparber (1939) bzw. (1940) 13–16. Handschriften: Hs. A (Zeissberg: A) = Neustift, Stiftsbibl., Cod. 293 (aus Neustift); Hs. B (Zeissberg: B) = Innsbruck, ULB Tirol, Cod. 216 (aus Schnals); Hs. C (Zeissberg: F) = München, BSB, Clm 11723 (aus Polling); Hs. D (Zeissberg: H) = Ed. Pez (nach einer jetzt verschollenen Hs. des Chorherrenstiftes Dürnstein); Hs. E = Klosterneuburg, Stiftsbibl., CCl 1241 (vgl. auch Schmölz 88) sowie fünf von Roschmann veranlasste Abschriften Fa–Fe, welche jeweils alle drei Viten enthalten (eine sechste von Roschmann veranlasste Hs. = nach Sparber Ff [= Klosterneu burg, Stiftsarchiv, K 105] enthält nur die Vita III). Drucke: Erstdruck in Pez 495–518, weitere Drucke s. Sparber 1911, 88 und Sparber 1939 bzw. 1940, 7–9. Zu den drei Hartmann-Viten: Vita I (Rec. I): um 1200, Verfasser wohl ein gewisser Hermannus (s. u.); Vita II (Rec. II): kürzere Fassung, unbekannter Chorherr in Neustift, 14. Jh.; Vita III: Abt Amandus Pachler von St. Peter in Salzburg, 1668, s. Sparber 1911, 86–88, Sparber 1939 bzw. 1940, 3–9. Zur Verfasserfrage vgl. Sparber 1940, 25–33: Entstehung der Handschrift nicht wie von Pez angenommen in Klosterneuburg, sondern in Neustift (oder Brixen), so Puell 180 Anm. 2. Als Verfasser von Puell, Anhang, 5 ein Hermannus, Verwalter des von Hartmann gegründeten Spitals in Brixen, 1200–1210 Propst von Neustift angenommen, nach Sparber 1940, 26–28 käme evtl. Konrad von Rodank in Frage, ca. 1180–1200 Propst von Neustift, 1200–1216 Bischof von Brixen, 1957 schließt sich Sparber (10 Anm. 5) jedoch der Meinung von Puell an. Cod. 293 nach Sparber 1940, 13 nicht das Original, sondern eine dem Original zeitlich nahestehende Abschrift. 2 (22r) Notiz aus Kloster Baumburg (Oberbayern) über die Weihe der dortigen Kirche (15. Jh.) (Ed. Zeissberg 454–455. MGH Scriptores XVII 438). Tit.: In monasterio Pawmburgensi Salczburgensis dyocesis ordinis sancti Augustini Canonico rum regularium sic scriptum invenitur. – Inc.: In nomine sancte et individue trinitatis et in h onore sancte Marie perpetue virginis et sancte Margarethe virginis et martiris omniumque celestium virtutum consecrata est hec ecclesia anno dominice incarnacionis millesimo centesimo quinqua gesimo sexto … – Expl.: … et a patre sanctissimo Brixinensis ecclesie Hartmanno etc. 3 (22r) Notiz über die Weihe der Kirche zu Seckau (13. Jh.) (Ed. Zeissberg 455). Schrift nach Zeissberg 13. Jh., nach Sparber 1940, 10 eher 16. oder 17. Jh. mit Nachahmung mittelalterlicher Schreibgewohnheiten. Inc.: Consecrata est Seccoviensis ecclesia a venerabili in Christo patre et domino Hartmanno Brixinensi episcopo circa annos (!) domini Mo Co XLo … – Expl.: … proxima dominica post nativitatis Marie que est patrona eiusdem ecclesie. (22v) leer. 4 (23r) Notiz über die Verwandtschaft der hl. Maria (13. Jh.) (Hauréau II 236va). Inc.: Ex testimoniis quatuor evangeliorum et exemplo Ieronimi contra Eheluinum (recte: Hel vidium) Sancta Maria mater domini et Maria mater Iacobi Alphei et Ioseph et mater filiorum Zebedei scilicet maioris Iacobi et Iohannis evangeliste tres sorores fuerunt … – Expl.: … Tres ergo viros Anna habuit Ioachim, Cleopam et Salome (!). Vgl. Leipzig, UB, Ms. 807, Bl. 166v–167r; Paris, BN, Ms. lat. 3751A, Bl. 28r. Zum Expl. vgl. Iacobus de Voragine, Legenda aurea, cap. 131 (Ed. Graesse 586). 5 (23r–v) 34 Verse über drei Heilige namens Joseph (15. Jh., von gleicher Hand wie Nr. 2). Tit.: Versus de meritis et vita trium sanctorum nomine Ioseph. – Inc.: Omnis homo certis sciat ex causis et apertis / quod tres sunt sancti quos Ioseph nomine legi … – Expl.: … Illi donetur per eos quod sanctificetur. 232 Cod. 308 (23v) Besitzvermerk (von gleicher Hand wie Nr. 2 und 5) (s. bei G). (23v) untere Blatthälfte, quer Anrufung an den hl. Hartmann: In nomine domini Iesu Christi Amen. O beate Hartmane (!) ora pro nobis ad dominum. (23v) Darunter radierter Besitzvermerk. Salve (?) … ego Chunradus (?) … Brix. (?) et de Mul bach (?) … nobilis … (Ir) leer bis auf Abklatsch des Besitzvermerks auf Bl. 23v. (Iv–IIv) leer. W. N. Cod. 308 THEOLOGISCHE SAMMELHANDSCHRIFT Perg. I, 228, I Bl. 220 × 150. Süddeutsch-österreichischer Raum, 14. Jh. B: Perg., Haar- und Fleischseite deutlich unterscheidbar, vereinzelt verschmutzt, insbes. am Lagenende bzw. -anfang, vereinzelt kleine Löcher und Risse, einzelne Blätter zum unteren Blattrand hin unregelmäßig. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 2.IV16 + (IV–1)23 + IV31 + (IV–1)38 + 2.IV54 + (IV–1)61 + IV69 + III75 + 15.IV195 + (IV–1)202 + 3.IV226 + I228 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nach satzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 18, 36, 59 und 196 herausgeschnitten (ohne Textverlust). Lage 1–26 am Lagenende Kustoden in römischen Zahlen; ab Lage 27 (Bl. 203ff.) Rekla manten, gerahmt, be- bzw. weggeschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Bl. 110 und 228 Spuren von ehem. Signakeln. S: Schriftraum 170/175 × 110. Zwei Spalten, von Tintenlinien gerahmt, zu 33–35 Zeilen auf Tintenlinierung. Zirkelstiche. Textualis. Vereinzelt Ergänzungen von derselben und von anderer zeitgenössischer Hand. Bl. 46rb, 49rb (möglicherweise J. Resch, 1716–1782, Priester und Historiker) und 219rb neuzeitliche Notizen von verschiedenen Händen. A: Rote Auszeichnungsstriche, Trennstriche und -punkte, Unter- und Durchstreichungen, Überschriften. Bl. 1r–201v am oberen Blattrand Angabe der jeweiligen Distinctio (auf dem Verso jeweils D, auf dem Recto römische Zahl, beginnend bei IIII) in Rot, tw. von kleinen roten Kreisen gerahmt. Ab Bl. 210ra auch rote Paragraphzeichen und am Blattrand vereinzelt rote Verweiszeichen sowie Zählung der Abschnitte in roten arabischen Ziffern. In der ersten Zeile sehr selten Oberlängen gedehnt. Am Beginn jeder Distinctio (s. Inhalt) jeweils Register mit meist ein-, selten zwei- bzw. dreizeiligen roten Lombarden und Seitenangaben in roten römischen Zahlen. Zwei- bis sechszeilige rote Lombarden, tw. mit Punktverdickungen, tw. mit Ausläufern, Bl. 52r Ausläufer in drei Dornblättern endend. Bl. 13vb sechszeilige blaue Initiale mit einfachem rotem Knospenfleuronée (im Binnenfeld und als Besatz) und Fadenausläufern. Bl. 35ra auf dem Bas-de-page Aus läufer mit zwei Medaillons, darin Knospenfleuronnée. Bl. 225va und 228rb am unteren Blattrand Repräsen tanten für Überschriften erhalten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Pap. leicht abgerieben und zerkratzt. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Gold druck SPECULUM EXEMPLORUM in Goldrahmung, im dritten Signaturschild (s. bei G). Schnitt rot gefärbt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II 2 E 1 (?, Tinte, radiert), II1 D 8 (Bleistift) und 308 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 308. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 10031. – Peintner, Stiftsbibliothek 118. – Katalog ULB Tirol IV 57, 471. – D. E. Mairho fer, Liber Lacteus. Eine unbeachtete Mirakel- und Exempelsammlung aus dem Zisterzienserkloster Stams [Codicologia 1]. Badenweiler 2009, 22 (ohne Nennung der Sign.). Cod. 308 1 1.1 233 (Ir–v) leer. (1ra–202ra) Lacteus liquor, unvollständig. Textbeginn in Dist. 4, cap. 7. Erhalten sind insgesamt 662 Exempla, die direkt im Anschluss um weitere 28 Exempla (s. Nr. 2 und 3) ergänzt sind. Ein Großteil der Exempla ist in der Legenda Aurea enthalten (hier abgeglichen mit Ed. Graesse; zuletzt ed. von G. P. Maggioni, Iacopo da Varazze. Legenda Aurea [Millenio Medievale 6, Testi 3]. Tavarnuzze [Firenze] 1998). Die vorliegende Sammlung stimmt trotz abweichender Anordnung der einzelnen Exempla vielfach mit Innsbruck, ULB Tirol, Cod. 494 (ausführliche Auflistung: Mairhofer, Liber Lacteus) überein. Da Cod. 308 zahlreiche zusätzliche Exempla ent hält, ist er wohl einem anderen Überlieferungsstrang zuzuordnen: Eine sehr große Überein stimmung besteht mit Heiligenkreuz, Stiftsbibl., Cod. 323, Bl. 55va–214vb (14. Jh.). Das Incipit mutile aus Cod. 308, 1ra findet sich in Heiligenkreuz, Stiftsbibl., Cod. 323 ebenfalls in Dist. 4, cap. 7 (hier Tit.: De eo qui domos sancti Laurencii abstulit). Abweichungen in Heiligenkreuz, Stiftsbibl., Cod. 323 (die Kapitelzahlen beziehen sich auf die Angaben in Cod. 308): Dist. 4: cap. 41 in Cod. 323 nach cap. 37. – Dist. 5 (in Cod. 323 o. Tit.): Prolog aus Cod. 308, Bl. 12rb fehlt in Cod. 323. Cap. 28–32 in Cod. 323 zw. cap. 35 und 36 eingeschoben. Cap. 66–69 in Cod. 323 Tit. falsch zugeordnet. Cap. 79 und 80 in Cod. 323 ein Exempel (ohne Untertei lung). – Dist. 6 (in Cod. 323 o. Tit.): Cap. 14 und 15 vertauscht. Zw. cap. 110 und 111 in Cod. 323 ein zusätzliches Exempel. Nach dem letzten cap. aus Cod. 308 in Cod. 323, Bl. 135va 12 zu sätzliche Exempla. – Dist. 7: Nach dem letzten Kapitel aus Cod. 308 in Cod. 323, Bl. 160vb ein zusätzliches Exempel. – Dist. 8: In Cod. 323, Bl. 162va Prolog. – Dist. 10: In Cod. 323, Bl. 198rb Prolog. Cap. 1–18, 28, 30, 52 und 55 nicht in Cod. 323. In Cod. 323 acht zusätzliche Exempla. Reihenfolge abweichend: sechs Exempla, cap. 27, 29, ein zusätzliches Exempel, cap. 20, 19, 23, 24, 21, 22, 25, 26, 31–45, ein zusätzliches Exempel, cap. 46–50, 53, 51, 54, 56, 57. Insgesamt enthalten die beiden Hs. von Dist. 4, cap. 7 bis Dist. 10, cap. 57 gleich viele Exempla, allerdings weichen 22 voneinander ab. Nach Dist. 10, cap. 57 folgen in Cod. 308 weitere 176 Exempla (Mairhofer Nr. 390–416 und 447–597). Zahlreiche Exempla auch in Neustift, Stifts bibl., Cod. 125 enthalten. Da von Heiligenkreuz, Stiftsbibl., Cod. 323 im Gegensatz zu Innsbruck, ULB Tirol, Cod. 494 keine genaue Aufschlüsselung vorliegt, werden im Folgenden gegebenenfalls die Nummern nach Mairhofer angeführt. Die Titel der einzelnen Exempla werden nur dann zitiert, wenn sie bei Mairhofer nicht aufscheinen oder von den dort angeführten Titeln abweichen. Text bisweilen von Drucken und Editionen abweichend, aber mit Mairhofer übereinstimmend. Incipit und Explicit werden nur dann angeführt, wenn sowohl von Drucken und Editionen als auch von Mairhofer abweichend. (1ra–12rb) Dist. 4: Miracula de martyribus. (1ra–vb) Laurentius: (1ra) Inc. mut.: ] panem invenit sed cum vix ad unius prandii esum tribus personis sufficere videntur Laurentius artifices suos nolens deficere … (Graesse Nr. 117 [112], 4). (1ra) Graesse Nr. 117 (112), 6. – Mairhofer Nr. 97. (1rb) Graesse Nr. 117 (112), 7 (S. 495, Z. 11– 32). – Mairhofer Nr. 98. (1vb) Tit.: De illis qui corpus eius viderunt (Graesse Nr. 117 [112], 7 [S. 495, Z. 33–S. 496, Z. 1]). (1vb–3ra) Thomas Cantuariensis: (1vb) Tit.: De sancto Thoma Cantuariense (!) archiepiscopo (Graesse Nr. 11, 1 [S. 67, Z. 9–12]). Inc.: Beatus Thomas sanctitatem suam ita subtiliter occulta bat ut salvo semper honestatis rigore … (2ra) Tit.: De excellencia huius martiris (Graesse Nr. 11, 2 [S. 68, Z. 2–7]). Expl.: … a domino servabatur. (2ra) Tit.: Qualiter ubi quando et a quibus passus fuerit (Graesse Nr. 11, 2 [S. 68, Z. 19–30]. – Mairhofer Nr. 99). (2rb) Tit.: De cantu ange lorum (Graesse Nr. 11, 3 [S. 68, Z. 30–S. 69, Z. 3]. – Mairhofer Nr. 100). (2va) Graesse Nr. 11, 3 (S. 69, Z. 7–13). – Mairhofer Nr. 101. (2va) Tit.: De efficacia aque sancti Thome (Graesse Nr. 11, 234 Cod. 308 4). (2vb) Tit.: De ave docta loqui quam nisus insequebatur (Graesse Nr. 11, 5). (2vb) Tit.: De quodam quem s. Thomas dilexit (Graesse 69 [Nr. 11, 6]). (3ra–va) Felix presbyter Romanus: (3ra) Tit.: De sancto Felice quem pueri peremerunt (Graesse Nr. 19 [S. 102, Z. 9–17]). (3ra) Tit.: Qualiter aranee ipsum abscondiverit (!) (Graesse Nr. 19 [S. 102, Z. 22–26]). – Inc.: Quadam vice cum predicans Felix predicaret et persecutor … (3ra) Tit.: De fratre eiusdem sancti (Graesse Nr. 19 [S. 102, Z. 30–S. 103, Z. 1]). (3rb) Tit.: De paganis qui eum tenere voluerunt (Graesse Nr. 19 [S. 103, Z. 5–8]). – Inc.: Quadam autem vice venerunt gentiles ut eum tenerent … (3rb) Tit.: Quot pontifices ydolorum baptizavit (Graesse Nr. 19 [S. 103, Z. 8–18]). (3va–b) Sebastianus: (3va) Graesse Nr. 23, 4. – Mairhofer Nr. 102. (3vb) Tit.: De maxima pesti lencia Ytalie (Graesse Nr. 23, 5 [S. 113, Z. 1–12]). – Expl.: … a Roma reliquie sunt delate. (3vb–4rb) Ignatius: (3vb) Tit.: Epistula Ignacii ad Mariam (Graesse Nr. 36, 1 [S. 155, Z. 3–10]). – Expl.: … ex te propter te et in te confirmeretur. (4ra) Tit.: Rescriptum gloriose virginis ad ipsum (Graesse Nr. 36, 1–2 [S. 155, Z. 10–27]). (4ra) Tit.: Quod Ignacius angelos audivit cantantes (Graesse Nr. 36, 3 [S. 155, Z. 27–30]). (4rb) Tit.: Quod Ihesu Christi nomen in corde scriptum habuit (Graesse Nr. 36, 4 [S. 157, Z. 28–36]). (4rb–va) Blasius: (4rb) Tit.: De sancto Blasio (Graesse Nr. 38 [S. 167, Z. 26–30]). (4va) Tit.: De porco quem lupus rapuit (Graesse Nr. 38 [S. 167, Z. 30–S. 168, Z. 3 und Z. 11–16]. – Mairhofer Nr. 103). (4va–b) Georgius: Tit.: Quod sanctus Georius (!) dux est Christianorum (Graesse Nr. 58 [56] [S. 264, Z. 26–34]). (4vb–5rb) Marcellinus papa (Ed.: A. Sorbelli, Corpus chronicorum Bononiensium [Rerum Itali carum scriptores. Raccolta degli storici italiani dal cinquecento al millecinquecento 28/1]. Città di Castello 1905, 206, Z. 18–207, Z. 33 [Cronaca A]. – Mairhofer Nr. 104). (5rb) Sancti Iohannes et Paulus: Tit.: De sanctis Iohanne et Paulo (Graesse Nr. 87 [82] [S. 367, Z. 5–12]). (5rb–6ra) Christophorus: Tit.: De sancto Christoforo insigne miraculum (Graesse Nr. 100 [95] [S. 431, Z. 29–S. 432, Z. 32]. – Mairhofer Nr. 22). (6ra–b) Mauricius et socii sui: (6ra) Graesse Nr. 141 (136), 2. – Mairhofer Nr. 105. (6rb) Tit.: De clerico ambicioso horrendum miraculum (Graesse Nr. 141 [136], 5. – Mairhofer Nr. 106). (6rb–7ra) Cosmas et Damianus (BHL S. 298, ohne Nr.): (6rb) Tit.: De sanctis Cosme (!) et Damiano (Graesse Nr. 143 [138], 2). (6va) Tit.: De viro qui peregre proficiscens uxorem eius conmendavit (Graesse Nr. 143 [138], 3. – Mairhofer Nr. 107). (6vb) Tit.: Quod coram cuidam viro absciderunt (Graesse Nr. 143 [138], 4). (7ra–10vb) Udo von Magdeburg: Tit.: De querela sancti Mauricii et duplici iudicio corporali et spirituali contra Udonem (Ed.: A. E. Schönbach, Studien zur Erzählungsliteratur des Mittelalters [SB Wien 144]. Wien 1901, 2–9. – Mairhofer Nr. 666). (10vb–12rb) Dionysius (Mairhofer Nr. 667). 1.2 (12rb–43va) Dist. 5: Miracula de episcopis. (12rb–vb) Prolog: Tit.: Incipit prologus Vo distinccionis. – Inc.: Virorum illustrium presulum scilicet ac doctorum preconia (!) sacrosanctam quinte huius distinccionis series continet et declarat nugas nimirum respuens … – Expl.: … et veritatis predicator principalissimus hec autem de eius laude breviter dicta sufficiant. Schlussschrift: Explicit distinccio IIIIa. (12vb–13vb) Register mit Kapitelzahlen. (13vb–17va) Augustinus: (13vb) Tit.: De conversione beati Augustini et ymno et domini l audamus prima distinccio (Graesse Nr. 124 [119], 1 [S. 552, Z. 36– S. 553, Z. 4 und S. 554, Z. 37–S. 555, Z. 13]). (14ra) Tit.: Quod tria a beato Ambrosio didicit (Graesse Nr. 124 [119], 1 [S. 555, Z. 19– 25]). (14rb) Tit.: Quod laudabat mori cupientes (Graesse Nr. 124 [119], 1 [S. 557, Z. 23–33]). Cod. 308 235 (14rb) Tit.: Quod mulieres sunt vitande (Graesse Nr. 124 [119], 1 [S. 560, Z. 39–S. 561, Z. 3]). – Inc.: Refert Gregorius in registro. Legitur quod sanctus Augustinus nec cum sorore … (14va) Tit.: Quare librum de civitate Dei composuerit (Graesse Nr. 124 [119], 1 [S. 558, Z. 24–33]). (14va) Tit.: De egro ab ipso sanato (Graesse Nr. 124 [119], 1 [S. 559, Z. 9–21]). – Inc.: Cum beatus Augustinus febribus laboraret et lecto decumberet … (14vb) Tit.: De transitu ipsius (Graesse Nr. 124 [119], 1 [S. 559, Z. 35–S. 560, Z. 4]). – Expl.: … floruit circa annum domini CCCCm XLm. (14vb) Tit.: De eo quod est illuminator ecclesie (Graesse Nr. 124 [119], 4 [S. 562, Z. 15– 20]. – Mairhofer Nr. 108). (15ra) Tit.: De corde sancti Augustini miraculum insigne (Mairhofer Nr. 109). (15va) Tit.: De sancto Bernhardo dormiens eum vidit (Graesse Nr. 124 [119], 4 [S. 562, Z. 20–26]). (15va) Tit.: De monacho qui digitum eius dedit (Graesse Nr. 124 [119], 5. – M airhofer Nr. 110). (15vb) Graesse Nr. 124 (119), 6. – Mairhofer Nr. 111. (16ra) Graesse Nr. 124 (119), 7. – Mairhofer Nr. 598. (16rb) Graesse Nr. 124 (119), 8. – Mairhofer Nr. 112. (16va) Tit.: De gloria et excellencia Augustini (Graesse Nr. 124 [119], 9. – Mairhofer Nr. 599). – Inc.: Fertur quoque quod dum vir quidam sancto Augustino valde devotus in spiritu raptus … (16va) Graesse Nr. 124 (119), 10. – Mairhofer Nr. 600. (16vb) Tit.: De proposito qui eum dilexit (Graesse Nr. 124 [119], 11. – Mairhofer Nr. 601). (17ra) Tit.: De CXL viris simul sanatis (Graesse Nr. 124 [119], 13. – Mairhofer Nr. 113). (17va–22vb) Gregorius: (17va) Graesse Nr. 46, 1. – Mairhofer Nr. 602. (17va) Graesse Nr. 46, 2. – Mairhofer Nr. 114. (17vb) Tit.: De pueris Anglicanis (Graesse Nr. 46, 3 [S. 190, Z. 1–29]. – Mairhofer Nr. 115). (18rb) Graesse Nr. 46, 4 (S. 191, Z. 10–18). – Mairhofer Nr. 603. (18rb) Graesse Nr. 46, 4 (S. 190, Z. 39–S. 191, Z. 1 und S. 191, Z. 37–S. 192, Z. 16). – Mairhofer Nr. 116. (18va) Graesse Nr. 46, 5 (S. 193, Z. 39–S. 194, Z. 6). – Mairhofer Nr. 604. (18vb) Graesse Nr. 46, 6 (S. 194, Z. 7–20). – Mairhofer Nr. 117. (19ra) Graesse Nr. 46, 6 (S. 194, Z. 20– 39). – Mairhofer Nr. 118. (19rb) Graesse Nr. 46, 7. – Mairhofer Nr. 605. (19va) Tit.: De fletu Gregorii (Graesse Nr. 46, 10 [S. 196, Z. 31–37]). (19va) Tit.: De egritudine eius (Graesse Nr. 46, 10 [S. 197, Z. 21–29]). (19vb) Tit.: Qualiter predicta intelligenda sint (Graesse Nr. 46, 10 [S. 197, Z. 4–21]). (20ra) Tit.: De iudicio Trayani (Graesse Nr. 46, 10 [S. 196, Z. 16–26 ]). (20rb) Tit.: De eodem (Graesse Nr. 46, 10 [S. 196, Z. 27–31]). (20rb) Tit.: De imperatore Mauricio (Graesse Nr. 46, 8 [S. 195, Z. 30–S. 196, Z. 1]. – Mairhofer Nr. 606). (20va) Graesse Nr. 46, 8 (S. 196, Z. 3–11). – Mairhofer Nr. 607. (20va) Tit.: De responsione angeli (Graesse Nr. 46, 9). (20vb) Tit.: De sancto Gregorio et matrona que panes ei obtulit (Graesse Nr. 46, 11–12. – Mairhofer Nr. 119). (21ra) Graesse Nr. 46, 13. – Mairhofer Nr. 120. (21rb) Graesse Nr. 46, 14. – Mairhofer Nr. 608. (21rb) Tit.: Quod officia ad missas composuit (Graesse Nr. 46, 15 [S. 199, Z. 3–10]). (21va) Tit.: Epytaphium tumbe eius (Graesse Nr. 46, 15 [S. 199, Z. 10–21]). (21vb) Graesse Nr. 46, 16. – Mairhofer Nr. 121. (22rb) Graesse Nr. 46, 17. – Mairhofer Nr. 122. (22vb–23rb) Ambrosius: (22vb) Tit.: De virtutibus beati Ambrosii (Graesse Nr. 57 [55], 1 [S. 250, Z. 16–22]). – Inc.: Ambrosius cum incunabulis in atrio pretorii esset positus et dormiret … (22vb) Tit.: Quod invitus factus est episcopus (Graesse Nr. 57 [55], 1 [S. 250, Z. 28–S. 251, Z. 13]. – Mairhofer Nr. 123). (23rb–va) Gregorius: Graesse Nr. 46, 19. – Mairhofer Nr. 609. (23va–26ra) Ambrosius: (23va) Graesse Nr. 57 (55), 2 (S. 251, Z. 21–32). – Mairhofer Nr. 610. (23vb) Graesse Nr. 57 (55), 2 (S. 251, Z. 32–S. 252, Z. 1). – Mairhofer Nr. 611. (24ra) Tit.: De heretico qui Dei angelum vidit (Graesse Nr. 57 [55], 2 [S. 252, Z. 12–16]). (24ra) Tit.: De aurus pice qui misit demones (Graesse Nr. 57 [55], 3 [S. 252, Z. 16–25]. – Mairhofer Nr. 612). (24ra) Tit.: De quodam cuius manus aruit (Graesse Nr. 57 [55], 3 [S. 252, Z. 25–S. 253, Z. 4]). (24rb) Graesse Nr. 57 (55), 4 (S. 253, Z. 4–8). – Mairhofer Nr. 613. (24rb) Tit.: De clauso palacio ne Ambrosius intraret (Graesse Nr. 57 [55], 4 [S. 253, Z. 8–14]). (24rb) Graesse Nr. 57 (55), 4 (S. 253 Z. 14–30). – Mairhofer Nr. 614. (24va) Tit.: De flagicioso quem tradidit dyabolo (Graes se Nr. 57 [55], 4 [S. 253, Z. 30–34]). – Expl.: … discrepare cepit. (24vb) Graesse Nr. 57 (55), 5. – Mairhofer Nr. 124. (25ra) Graesse Nr. 57 (55), 6 (S. 255, Z. 15–21). – Mairhofer Nr. 615. 236 Cod. 308 (25ra) Tit.: Nota quod in multis fuit commendabilis (Graesse Nr. 57 [55], 7–8. – Mairhofer Nr. 616). (26ra–29va) Martinus ep.: (26ra) Tit.: De virtutibus sancti Martini (Graesse Nr. 166 [161] [S. 741, Z. 29–S. 742, Z. 33]. – Mairhofer Nr. 125). (26rb) Graesse Nr. 166 (161) (S. 742, Z. 33– 39). – Mairhofer Nr. 126. (26va) Graesse Nr. 166 (161) (S. 743, Z. 35–S. 744, Z. 10). – Mairhofer Nr. 127. (26vb) Tit.: Qualiter tercium mortuum suscitavit (Graesse Nr. 166 [161] [S. 744, Z. 10– 16]). (26vb) Tit.: Quod ei obediebant insensibilia (Graesse Nr. 166 [161] [S. 744, Z. 16–25]). – Expl. erweitert: … magna successit qui conversus fidem suscepit. (27ra) Tit.: Quod ei obediebant vegetabilia (Graesse Nr. 166 [161] [S. 744, Z. 25–33]). (27ra) Tit.: Quod ei obediebant animalia (Graesse Nr. 166 [161] [S. 744, Z. 33–S. 745, Z. 5]. – Mairhofer Nr. 617). (27rb) Tit.: Quod multe fuerit humilitatis beatus Martinus (Graesse Nr. 166 [161] [S. 745, Z. 5–20]. – Mairhofer Nr. 128). (27va) Graesse Nr. 166 (161) (S. 745, Z. 21–31). – Mairhofer Nr. 129. (27va) Tit.: Multe fuit paciencie (Graesse Nr. 166 [161] [S. 745, Z. 31–S. 746, Z. 4]). (27vb) Tit.: Multe fuit compassionis (Graesse Nr. 166 [161] [S. 746, Z. 24–29]). (27vb) Tit.: Multe fuit largitatis (Graes se Nr. 166 [161] [S. 746, Z. 29–S. 747, Z. 16]. – Mairhofer Nr. 130). (28rb) Tit.: Multe fuit potestatis (Graesse Nr. 166 [161] [S. 747, Z. 17–26]). – Inc.: Multe fuit potestatis erga demones pellendos non solum ex hominibus … – Expl.: … ad armentum suum revertitur. (28rb) Tit.: Multe fuit subtilitatis (Graesse Nr. 166 [161] [S. 747, Z. 26–S. 748, Z. 6]). – Inc.: Multe subtilitatis fuit erga demones cognoscendos. Quadam vice dyabolus in forma regis purpura … (28va) Tit.: Quod moriens dyabolum vidit (Graesse Nr. 166 [161] [S. 748, Z. 6–35]). – Inc.: Cum Martinus pergeret ad Condacensem (!) dyocesin causa sedande discordie vidit mergos in flumine (gestrichen: piscem) piscibus insidiantes … (28vb) Graesse Nr. 166 (161) (S. 749, Z. 2–14). – Mairhofer Nr. 131. (29ra) Graesse Nr. 166 (161) (S. 749, Z. 14–21). Inc. gegenüber Ed. leicht abweichend, übereinstimmend mit Mairhofer Nr. 618. (29rb) Tit.: De eius exequiis et beato Ambrosio (Graes se Nr. 166 [161] [S. 749, Z. 22–34]. – Mairhofer Nr. 619). – Inc.: Sanctus Ambrosius eo die missam celebrans super altare … (29rb) Graesse Nr. 166 (161) (S. 750, Z. 7–20). – Mairhofer Nr. 620. (29va–33va) Nicolaus: (29va) Tit.: De virtutibus sancti Nycolay (Graesse Nr. 3, 1 [S. 23, Z. 2–16]). (29vb) Tit.: Quod nautis in periculo subvenit (Graesse Nr. 3, 3 [S. 23, Z. 9–S. 24, Z. 36). (30ra) Tit.: Quod famem a suis repulit (Graesse Nr. 3, 4. – Mairhofer Nr. 132). (30ra) Graesse Nr. 3, 6 (S. 25, Z. 11–24). – Mairhofer Nr. 133. (30rb) Graesse Nr. 3, 6 (S. 25, Z. 6–11 und Z. 24–26). – Mairhofer Nr. 134. (31ra) Graesse Nr. 3, 8. – Mairhofer Nr. 135. (31rb) Tit.: De ymagine beati Nycolai (Graesse Nr. 3, 9. – Mairhofer Nr. 136). (31va) Graesse Nr. 3, 10. – Mairhofer Nr. 621. (31vb) Tit.: De cypho aureo et de quodam nobili (Graesse Nr. 3, 11. – Mairhofer Nr. 622). (32ra) Tit.: De divite sibi devoto (Graesse Nr. 3, 12). (32rb) Mairhofer Nr. 623. (32vb) Tit.: De quodam fatuo sapiente iocosum (Mairhofer Nr. 137). (33rb–va) Pelagius papa: Tit.: De quodam priore ab eo plagato (Graesse Nr. 181 [176], 4 [S. 841, Z. 32–S. 842, Z. 12]). (33va–34va) Remigius: (33va) Graesse Nr. 16 (S. 96, Z. 5–16). – Mairhofer Nr. 138. Expl.: … autem omnia per ordinem facta sunt. (33vb) Tit.: De vino augmentato (Graesse Nr. 16 [S. 96, Z. 18–25]). – Inc.: Quodam tempore cum Remigius factus est Remensis archiepiscopus et in domo cuiusdam … (33vb) Tit.: Quod Clodoneum (!) regem baptizavit (Graesse Nr. 16 [S. 96, Z. 25–35]). (34ra) Graesse Nr. 16 (S. 96, Z. 35–S. 97, Z. 28). – Mairhofer Nr. 139. (34va–b) Hilarius: Graesse Nr. 17 (S. 98, Z. 7–8 und S. 99 Z. 3–24). – Mairhofer Nr. 624. (34vb–37va) Basilius: (34vb) Graesse Nr. 26, 1 (S. 121, Z. 1–7). – Mairhofer Nr. 625. (34vb) Tit.: De heremita qui eum despexit (Graesse Nr. 26, 1 [S. 121, Z. 7–20]. – Mairhofer Nr. 625 [Forts.]). (35ra) Tit.: De quodam alio et cauda catte (Graesse Nr. 26, 2). (35ra) Tit.: De ma gnimitate Basilii in corripiendo (Graesse Nr. 26, 3). – Expl. erweitert: … ecclesia catholicis sicut in tripartita hystoria dicitur. (35va) Tit.: De famulo qui se dedit dyabolo (Graesse Nr. 26, 5. – Cod. 308 237 Mairhofer Nr. 140). (36vb) Graesse Nr. 26, 6. – Mairhofer Nr. 626. (37rb) Graesse Nr. 26, 7. – Mairhofer Nr. 627. (37va–40rb) Iohannes Elemosinarius: (37va) Graesse Nr. 27, 1 (S. 126, Z. 19–28). – Mairhofer Nr. 141. (37va) Graesse Nr. 27, 1 (S. 126, Z. 28–S. 127, Z. 1). – Mairhofer Nr. 142. (37vb) Graes se Nr. 27, 1 (S. 127, Z. 1–24). – Mairhofer Nr. 143. (38ra) Graesse Nr. 27, 1 (S. 127, Z. 25–39). – Mairhofer Nr. 628. (38rb) Graesse Nr. 27, 2 (S. 127, Z. 39–S. 128, Z. 28). – Mairhofer Nr. 629. (38va) Tit.: De monacho qui cum meretricibus pernoctebat (Graesse Nr. 27, 3. – Mairhofer Nr. 144). (39rb) Graesse Nr. 27, 4. – Mairhofer Nr. 630. (39rb) Tit.: De quodam tabernario et suo nepote (Graesse Nr. 27, 6). (39va) Tit.: De precioso coopertorio cuiusdam divitis (Graesse Nr. 27, 8. – Mairhofer Nr. 145). (39vb) Graesse Nr. 27, 9. – Mairhofer Nr. 631. (40ra) Tit.: De magna eius paciencia (Graesse Nr. 27, 10). (40ra) Graesse Nr. 27, 11. – Mairhofer Nr. 146. (40ra) Tit.: De iuvene qui rapuit monialem (Graesse Nr. 27, 12 [S. 132, Z. 5–13]). (40rb–va) Leo papa: Graesse Nr. 88 (83) (S. 368, Z. 2–12). – Mairhofer Nr. 147. (40va–41rb) Germanus ep.: (40va) Graesse Nr. 107 (102) (S. 448, Z. 27–S. 449, Z. 12). – Mairhofer Nr. 148. (40va) Graesse Nr. 107 (102) (S. 449, Z. 22–30). – Mairhofer Nr. 149. (40vb) Graesse Nr. 107 (102) (S. 449, Z. 30–S. 450, Z. 11). – Mairhofer Nr. 150. (40vb) Graesse Nr. 107 (102) (S. 450, Z. 11–24). – Mairhofer Nr. 632. (41ra) Graesse Nr. 107 (102) (S. 450, Z. 33–39). – Mairhofer Nr. 151. (41rb) Graesse Nr. 107 (102) (S. 451, Z. 7–16). – Mairhofer Nr. 633. (41rb–va) Lupus: (41rb) Tit.: De sancto Lupo Senonensi archiepiscopo (Graesse Nr. 128 [123] [S. 579, Z. 7–12]). – Inc.: Cum sancto Lupo Senonensi archiepiscopo omnes qui de curia … (41va) Tit.: De eius convivio et vini defectu (Graesse Nr. 128 [123] [S. 579, Z. 3–7]). – Expl. erweitert: … pre foribus nunciavit et dixit Ecce quam verum est. Date et dabitur vobis etc. (41va) Tit.: Quod dyabolum inclusit (Graesse Nr. 128 [123] [S. 580, Z. 4–9]). (41va–42ra) Iohannes Chrysostomus: (41va) Graesse Nr. 138 (131) (S. 616, Z. 25–29). – Mair hofer Nr. 152. (41vb) Graesse Nr. 138 (131) (S. 616, Z. 30–34). – Mairhofer Nr. 153. (41vb) Graesse Nr. 138 (131) (S. 616, Z. 34–S. 617, Z. 5). – Mairhofer Nr. 154. (42ra) Tit.: De monacho sancto in episcopum electo (Graesse 138 [131] [S. 618, Z. 11–17]). – Inc.: Eo tempore in Lamon sanctissimus monachus … (42ra–43va) Patricius: (42ra) Graesse Nr. 50 (49) (S. 213, Z. 17–23). – Mairhofer Nr. 155. (42ra) Tit.: Quod sanctus Patricius omnes cruces adorabat (Graesse Nr. 50 [49] [S. 213, Z. 23–29]). – Expl.: … fecit eam inde tolli. (42rb) Graesse Nr. 50 (49) (S. 213, Z. 29–S. 216, Z. 13). – Mair hofer Nr. 156. (43va) Schlussschrift: Explicit distinccio Va. 1.3 (43va–72vb) Dist. 6: Miracula de monachis. (43va–44vb) Register mit Kapitelzahlen. (44vb–45va) Paulus eremita: (44vb) Graesse Nr. 15 (S. 94, Z. 9–28). – Mairhofer Nr. 157. (45ra) Graesse Nr. 15 (S. 94, Z. 28–S. 95, Z. 18). – Mairhofer Nr. 158. (45va) Tit.: De eius transitu ad celos (Graesse Nr. 15 [S. 95, Z. 18–27]. – Mairhofer Nr. 159). (45va–47rb) Antonius: (45va) Tit.: De virtutibus sancti Anthonii (Graesse Nr. 21, 1 [S. 104, Z. 8–12). – Mairhofer Nr. 160. (45va) Graesse Nr. 21, 1 (S. 104, Z. 12–17). – Mairhofer Nr. 161. (45vb) Graesse Nr. 21, 1 (S. 104, Z. 17–32). – Mairhofer Nr. 162. (46ra) Tit.: Quod martirium desideravit (Graesse Nr. 21, 1 [S. 104, Z. 32–S. 105, Z. 1]). (46ra) Tit.: De disco argenteo (Graes se Nr. 21, 2 [S. 105, Z. 1–8]). (46ra) Tit.: De laqueis dyaboli (Graesse Nr. 21, 2 [S. 105, Z. 9–12]). – Inc.: Quadam vice cum in spiritu temptatus esset totum mundum laqueis sese invicem … (46rb) Tit.: Quod dyabolum vidit (Graesse Nr. 21, 3 [S. 105, Z. 18–22]). (46rb) Graes se Nr. 21, 3 (S. 105, Z. 22–29). – Mairhofer Nr. 634. (46rb) Tit.: Quod cum fratribus quinque gaudebat (Graesse Nr. 21, 3 [S. 105, Z. 29–36]). (46rb) am unteren Blattrand Notiz (16. Jh.?), radiert: 1544 […] IIII […] jar. (46va) Graesse Nr. 21,4 (S. 106, Z. 2–10). – Mairhofer Nr. 635. (46va) Graesse Nr. 21, 5 (S. 106, Z. 19–23). – Mairhofer Nr. 637. (46vb) Graesse Nr. 21, 5 238 Cod. 308 (S. 106, Z. 10–18). – Mairhofer Nr. 636. (46vb) Tit.: Quod angelus eum docuit (Graesse Nr. 21, 5 [S. 106, Z. 23–27]. – Mairhofer Nr. 638). (46vb) Graesse Nr. 21, 5 (S. 106, Z. 27–36). – Mairhofer Nr. 639. (47ra–b) Tit.: De verbo salutis et suco (Graesse Nr. 21, 6 [S. 107, Z. 20–28]). (47rb–48ra) Macarius: (47rb) Tit.: De sancto Machario et eius miraculis (Graesse Nr. 18 [S. 100, Z. 16–25]. – Mairhofer Nr. 640). (47rb) Tit.: Quod humilitate dyabolum vicit (Graesse Nr. 18 [S. 100, Z. 25–S. 101, Z. 1]. – Mairhofer Nr. 641). (47va) Tit.: Quod corpus suum afflixit (Graes se Nr. 18 [S. 101, Z. 5–8]. – Mairhofer Nr. 163). – Inc.: Dum temptaciones dyaboli ipsum nimio molestarent … (47va) Graesse Nr. 18 (S. 101, Z. 8–23). – Mairhofer Nr. 164. (47vb) Tit.: De ca pite pagani (Graesse Nr. 18 [S. 101, Z. 23–32]. – Mairhofer Nr. 165). (47vb) Tit.: Quod dyabolus harundines collegit (Graesse Nr. 18 [S. 101, Z. 32–36]). (48ra) Tit.: De fratre qui multis prodesse voluit (Graesse Nr. 18 [S. 101, Z. 36–S. 102, Z. 4]). (48ra) Tit.: De culice (!) et reprehensione (Graesse Nr. 18 [S. 102, Z. 4–7]). (48ra–vb) Pastor: (48ra) Tit.: De abbate Pastore (Graesse Nr. 175 [170] [S. 803, Z. 15–26]. – Mairhofer Nr. 166). (48rb) Graesse Nr. 175 (170) (S. 804, Z. 21–27). – Mairhofer Nr. 167. (48rb) Tit.: Quod ira saluti sit contraria (Graesse Nr. 175 [170] [S. 804, Z. 27–31]). (48va) Tit.: Qualis debeat esse monachus (Graesse Nr. 175 [170] [S. 804, Z. 31–37]). (48va) Tit.: Notabile de cogi tacionibus (Graesse Nr. 175 [170] [S. 805, Z. 2–6]. – Mairhofer Nr. 642). (48va) Graesse Nr. 175 (170) (S. 805, Z. 6–11). – Mairhofer Nr. 643. (48vb–49ra) Iohannes abb.: (48vb) Tit.: De abbate Iohanne (Graesse Nr. 176 [171] [S. 805, Z. 13–22]). – Inc.: Iohannes abbas cum quendam qui per XL annos in heremo habitaverat in terrogasset … (48vb) Graesse Nr. 176 (171) (S. 805, Z. 22–S. 806, Z. 6). – Mairhofer Nr. 168. (49ra–va) Moyses abb.: (49ra) Graesse Nr. 177 (172) (S. 806, Z. 8–19). – Mairhofer Nr. 169. (49rb) Tit.: Notandum (Graesse Nr. 177 [172] [S. 806, Z. 19–22]). (49rb) Graesse Nr. 177 (172) (S. 806, Z. 22–26). – Mairhofer Nr. 644. (49rb) Graesse Nr. 177 (172) (S. 806, Z. 27–S. 807, Z. 2). – Mairhofer Nr. 645. (49va) Graesse Nr. 177 (172) (S. 807, Z. 3–10). – Mairhofer Nr. 646. (49va–50vb) Arsenius abb.: (49va) Tit.: Quod taciturnitas est salutis inicium (Graesse Nr. 178 [163] [S. 807, Z. 11–14]). (49vb) Graesse Nr. 178 (163) (S. 807, Z. 14–26). – Mairhofer Nr. 170. (49vb) Graesse Nr. 178 (163) (S. 807, Z. 26–S. 808, Z. 2). – Mairhofer Nr. 171. (50ra) Graesse Nr. 178 (163) (S. 808, Z. 3–22). – Mairhofer Nr. 172. (50rb) Tit.: Item unde supra (Graesse Nr. 178 [163] [S. 808, Z. 22–26]). (50rb) Tit.: Quod corpus mulieris ignis est (Graesse Nr. 178 [163] [S. 808, Z. 26–30]). (50rb) Tit.: De gracia lacrimarum (Graesse Nr. 178 [163] [S. 808, Z. 30–37]. – Mairhofer Nr. 647). (50va) Graesse Nr. 178 (163) (S. 808, Z. 37–S. 809, Z. 7). – Mairhofer Nr. 648. (50va) Graesse Nr. 178 (163) (S. 809, Z. 7–23). – Mairhofer Nr. 649. (50vb–51va) Agathon abb.: (50vb) Tit.: De sancto qui lapidem per triennium in ore portavit (Graesse Nr. 179 [174] [S. 809, Z. 27–S. 810, Z. 1]). – Expl.: … cum leditur foras egreditur. (51ra) Tit.: Quod maximus labor est orare (Graesse 179 [174] [S. 810, Z. 1–6]). (51ra) Tit.: Quod iracundi Deo non placent (Graesse 179 [174] [S. 810, Z. 6–10]). (51ra) Graesse Nr. 179 (174) (S. 810, Z. 11–18). – Mairhofer Nr. 650. (51rb) Graesse Nr. 179 (174) (S. 810, Z. 18–26). – Mairhofer Nr. 651. (51rb) Tit.: De iniuria non retinenda (Graesse 179 [174] [S. 810, Z. 28–30]). (51rb) Graesse Nr. 179 (174) (S. 810, Z. 30–S. 811, Z. 3). – Mairhofer Nr. 173. (51va–53ra) Ieronimus: (51va) Graesse 146 (141) (S. 654, Z. 1–25). – Mairhofer Nr. 174. (51vb) Graesse 146 (141) (S. 654, Z. 25–S. 655, Z. 19). – Mairhofer Nr. 175. (52ra) Tit.: Quod in Beth leem habitabat (Graesse 146 [141] [S. 655, Z. 19–31]). (52rb) Graesse 146 (141) (S. 655, Z. 31–S. 657, Z. 13). – Mairhofer Nr. 176. (53ra–57va) Benedictus: (53ra) Graesse 49 (48), 1 (S. 204, Z. 17–205). – Mairhofer Nr. 177. (53rb) Graesse 49 (48), 1 (S. 205, Z. 2–11). – Mairhofer Nr. 178. (53rb) Graesse Nr. 49 (48), 2. – Mairhofer Nr. 179. (53va) Graesse Nr. 49 (48), 3 (S. 206, Z. 1–14). – Mairhofer Nr. 180. (53vb) Tit.: De ferro quod in profundum cecidit (Graesse 49 [48], 3 [S. 206, Z. 25–29]). (53vb) Graesse Nr. 49 (48), 3 (S. 206, Z. 29–39). – Mairhofer Nr. 181. (54ra) Tit.: De corvo et pane Cod. 308 239 infecto (Graesse 49 [48], 3 [S. 206, Z. 39–S. 207, Z. 24]. – Mairhofer Nr. 652). (54va) Tit.: Quod dyabolus contra eum seviebat (Graesse Nr. 49 [48], 3 [S. 207, Z. 27–32]). – Inc.: Quadam vice antiquus hostis corporalibus eius … (54va) Graesse Nr. 49 (48), 3 (S. 207, Z. 32–S. 208, Z. 7). – Mairhofer Nr. 182. (54vb) Graesse Nr. 49 (48), 4. – Mairhofer Nr. 653. (55ra) Tit.: Quod spiritum prophecie habuit (Graesse Nr. 49 [48], 5). (55ra) Graesse Nr. 49 (48), 6. – Mairhofer Nr. 183. (55rb) Graesse Nr. 49 (48), 7 (S. 208, Z. 37–S. 209, Z. 7). – Mairhofer Nr. 654. (55va) Graesse 49 (48), 7 (S. 209, Z. 7–14). – Mairhofer Nr. 655. (55va) Tit.: De monachis monasterium con struentibus (Graesse 49 [48], 11). – Expl.: … audistis omnia ordinate. (55vb) ‹Ps.-Gregorius Magnus›: Dialogi, Lib. 2, cap. 12 (Ed.: F. Clark, The Pseudo-Gregorian dialogues 1. Leiden 1987. – Mairhofer Nr. 656). (56ra) Graesse Nr. 49 (48), 12. – Mairhofer Nr. 657. (56rb) Tit.: De fratre qui sine licencia exivit (Graesse Nr. 49 [48], 13. – Mairhofer Nr. 243). (56rb) Graesse Nr. 49 (48), 14. – Mairhofer Nr. 658. (56va) Graesse Nr. 49 (48), 15. – Vgl. Mairhofer Nr. 659. (56vb) Graesse Nr. 49 (48), 16. – Mairhofer Nr. 184. (57ra) Tit.: Quod totum mundum simul et semel vidit (Graesse Nr. 49 [48], 17). (57rb) Graesse Nr. 49 (48), 18. – Mairhofer Nr. 660. (57va–64rb) Bernardus: (57va) Tit.: De virtutibus beati Bernhardi doctoris egregii (Graesse Nr. 120 [115] [S. 528, Z. 16–22]. – Mairhofer Nr. 661). (57va) Tit.: De dolore capitis eius in puericia (Graesse Nr. 120 [115] [S. 528, Z. 23–27]). (57vb) Tit.: Quod puer Ihesus ei apparuit (Graesse Nr. 120 [115] [S. 528, Z. 27–32]). (57vb) Tit.: De puella que in lectum eius cecidit (Graesse Nr. 120 [115] [S. 529, Z. 7–13]). (57vb) Graesse Nr. 120 (115) (S. 529, Z. 13–29). – Mairhofer Nr. 663. (58rb) Graesse Nr. 120 (115) (S. 529, Z. 29–S. 530, Z. 9). – Mairhofer Nr. 185. (58va) Tit.: De ingressu eius in Cystercium (Graesse Nr. 120 [115] [S. 530, Z. 9–19]). (58vb) Tit.: De mira eius conversione sacione (!) (Graesse Nr. 120 [115] [S. 530, Z. 19–25]). (58vb) Graesse Nr. 120 (115) (S. 530, Z. 25–S. 531, Z. 10). – Mairhofer Nr. 186. (59ra) Tit.: De eius studiis (Graesse Nr. 120 [115] [S. 531, Z. 10–13]). (59rb) Tit.: De eius mundicia corporali (Graesse Nr. 120 [115] [S. 531, Z. 19–23]). (59rb) Tit.: De canonico qui ad maxillam eum per cussit (Graesse Nr. 120 [115] [S. 532, Z. 4–17]). – Inc.: Quidam canonicus ad eum venit et ut ad monachum eum reciperet … (59va) Graesse Nr. 120 (115) (S. 532, Z. 22–S. 533, Z. 2). – Mair hofer Nr. 187. (59vb) Graesse Nr. 120 (115) (S. 533, Z. 2–12). – Mairhofer Nr. 188. (60ra) Tit.: Quod fratres pro eo oraverunt (Graesse Nr. 120 [115] [S. 533, Z. 14–18]). – Expl. erweitert: … sinite me abire. Noviciis autem intrare volentibus dicere consueverat. Si ad ea que intus sunt festinatis hic foris dimittite corpora que a seculo attulistis. Soli spiritus ingrediantur caro autem non prodest quicquam. (60rb) Tit.: Quod tota die iuxta lacum pergens eum non vidit (Graesse Nr. 120 [115] [S. 533, Z. 24–S. 534, Z. 7]). (60va) Tit.: De temptacione eius cum predicaret qualiter eum vicit (Graesse Nr. 120 [115] [S. 534, Z. 7–14]). (60va) Graesse Nr. 120 (115) (S. 534, Z. 15–30). – Mairhofer Nr. 662. (60vb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 2, cap. 15 (PL 185, 1324D–1325C. – Mairhofer Nr. 664). (61rb) ‹Herbertus Clarae vallensis›: De miraculis, Lib. 2, cap. 16 (PL 185, 1325C–1326C. – Mairhofer Nr. 665). (61vb) Graesse Nr. 120 (115) (S. 534, Z. 30–S. 535, Z. 8). – Mairhofer Nr. 189. (62ra) Tit.: De epistula quam sub divo scripsit (Graesse Nr. 120 [115] [S. 535, Z. 8–17]). – Inc.: Frater Ruodbertus eius monachus et secundum carnem … (62ra) Tit.: De multis quas excommunicavit (Graesse Nr. 120 [115] [S. 535, Z. 17–20]). (62rb) Tit.: De obsessa quam liberavit (Graesse Nr. 120 [115] [S. 535, Z. 20–S. 536, Z. 10]). – Mairhofer Nr. 190. (62vb) Graesse Nr. 120 (115) (S. 536, Z. 10–26). – Mairhofer Nr. 191. (63ra) Graesse Nr. 120 (115) (S. 536, Z. 27–S. 537, Z. 2). – Mairhofer Nr. 192. (63rb) Graesse Nr. 120 (115) (S. 537, Z. 3–12). – Mairhofer Nr. 193. (63va) Graesse Nr. 120 (115) (S. 537, Z. 13–24). – Mairhofer Nr. 194. (63va) Graesse Nr. 120 (115) (S. 537, Z. 24–33). – Mairhofer Nr. 195. (63vb) Graesse Nr. 120 (115) (S. 537, Z. 33–S. 538, Z. 2). – Mairhofer Nr. 196. (63vb) Graesse Nr. 120 (115) (S. 538, Z. 2–12). – Mairhofer Nr. 197. (64ra) Graesse Nr. 120 (115) (S. 538, Z. 12–22). – Mairhofer Nr. 198. (64rb–va) Aegidius: Tit.: De sancto Egidio abbate et Karulo imperatore (Graesse Nr. 130 [125] [S. 584, Z. 2–14]. – Mairhofer Nr. 199). 240 Cod. 308 (64va–b) Pelagius papa: Graesse Nr. 181 (176), 4 (S. 833, Z. 13–S. 834, Z. 2). – Mairhofer Nr. 200. (64vb–68rb) Dominicus: (64vb) Graesse Nr. 113 (108) (S. 467, Z. 10–29). – Mairhofer Nr. 201. (65ra) Graesse Nr. 113 (108) (S. 468, Z. 26–S. 469, Z. 11). – Mairhofer Nr. 202. (65va) Tit.: Quod Petrus baculum Paulus sibi librum tradidit (Graesse Nr. 113 [108] [S. 469, Z. 11–18]. – Mairhofer Nr. 203). (65va) Graesse Nr. 113 (108) (S. 470, Z. 3–28). – Mairhofer Nr. 204. (66ra) Graesse Nr. 113 (108) (S. 470, Z. 28–38). – Mairhofer Nr. 205. (66rb) Graesse Nr. 113 (108) (S. 470, Z. 38–S. 471, Z. 13). – Mairhofer Nr. 206. (66rb) Graesse Nr. 113 (108) (S. 471, Z. 13– 19). – Mairhofer Nr. 207. (66va) Graesse Nr. 113 (108) (S. 471, Z. 28–39). – Mairhofer Nr. 208. (66vb) Tit.: De iuvenibus ignotis qui ei panes detulerunt (Graesse Nr. 113 [108] [S. 473, Z. 32–S. 474, Z. 5]. – Mairhofer Nr. 209). (66vb) Graesse Nr. 113 (108) (S. 474, Z. 5–10). – Mairhofer Nr. 210. (67ra) Graesse Nr. 113 (108) (S. 476, Z. 25–31). – Mairhofer Nr. 211. (67ra) Graesse Nr. 113 (108) (S. 477, Z. 20–S. 478, Z. 18). – Mairhofer Nr. 212. (67vb) Graesse Nr. 113 (108) (S. 478, Z. 19–35). – Mairhofer Nr. 213. (67vb) Tit.: Quod gratus Deo locus est in quo corpus viri Dei requiescit (Graesse Nr. 113 [108] [S. 482, Z. 13–S. 483, Z. 8]. – Mairhofer Nr. 214). (68rb–72vb) Franciscus: (68rb) Graesse Nr. 149 (144) (S. 663, Z. 6–22). – Mairhofer Nr. 215. (68va) Tit.: Quod carnalis frater suus eum derisit (Graesse Nr. 149 [144] [S. 664, Z. 2–6]. – Mairhofer Nr. 216). (68vb) Graesse Nr. 149 (144) (S. 664, Z. 6–10 und 16–20). – Mairhofer Nr. 217. (68vb) Tit.: Quod ei adheserunt (Graesse Nr. 149 [144] [S. 664, Z. 10–16]. – Mairhofer Nr. 218). (69ra) Tit.: De ypocrita (Graesse Nr. 149 [144] [S. 664, Z. 20–29]. – Mairhofer Nr. 219). (69rb) Graesse Nr. 149 (144) (S. 664, Z. 29–35). – Mairhofer Nr. 220. (69rb) Graesse Nr. 149 (144) (S. 665, Z. 15–25). – Mairhofer Nr. 221. (69va) Graesse Nr. 149 (144) (S. 665, Z. 8–15). – Mairhofer Nr. 222. (69va) Graesse Nr. 149 (144) (S. 665, Z. 25–32). – Mairhofer Nr. 223. (69vb) Graesse Nr. 149 (144) (S. 666, Z. 33–S. 667, Z. 3). – Mairhofer Nr. 224. (70ra) Graesse Nr. 149 (144) (S. 666 Z. 15–33). – Mairhofer Nr. 225. (70rb) Tit.: Quod demones provocavit (Graesse Nr. 149 [144] [S. 667, Z. 4–11]. – Mairhofer Nr. 226). (70va) Tit.: De eius excellenti gloria (Graesse Nr. 149 [144] [S. 667, Z. 11–20]. – Mairhofer Nr. 227). (70va) Tit.: Quod stigmata Christi habuit (Graesse Nr. 149 [144] [S. 667, Z. 20–26]. – Mairhofer Nr. 228). (70vb) Graesse Nr. 149 (144) (S. 668, Z. 18–30). – Mairhofer Nr. 229. (70vb) Graesse Nr. 149 (144) (S. 668, Z. 34–39). – Mairhofer Nr. 230. (71ra) Graesse Nr. 149 (144) (S. 668, Z. 39–S. 669, Z. 8). – Mairhofer Nr. 231. (71rb) Graesse Nr. 149 (144) (S. 669, Z. 31–36). – Mairhofer Nr. 232. (71rb) Graesse Nr. 149 (144) (S. 670, Z. 19–27). – Mairhofer Nr. 233. (71va) Graesse Nr. 149 (144) (S. 670, Z. 27–31). – Mairhofer Nr. 234. (71va) Graesse Nr. 149 (144) (S. 670, Z. 7–14). – Mairhofer Nr. 235. (71vb) Graesse Nr. 149 (144) (S. 670, Z. 32–S. 671, Z. 1). – Mairhofer Nr. 236. (71vb) Graesse Nr. 149 (144) (S. 671, Z. 13–25). – Mairhofer Nr. 237. (72ra) Graesse Nr. 149 (144) (S. 671, Z. 29–35). – Mairhofer Nr. 238. (72rb) Graesse Nr. 149 (144) (S. 671, Z. 39–S. 672, Z. 8). – Mairhofer Nr. 239. (72rb) Tit.: Quod celos ascendit (Graesse Nr. 149 [144] [S. 672, Z. 10–15]. – Mairhofer Nr. 240). (72va) Graesse Nr. 149 (144) (S. 673, Z. 6–15). – Mairhofer Nr. 241. (72vb) Schlussschrift: Explicit distinccio VI. 1.4 (72vb–92ra) Dist. 7: Miracula de sanctis virginibus. (72vb–73ra) Register mit Kapitelzahlen. (73ra–76vb) Maria Magdalena: (73ra) Graesse Nr. 96 (90), 1–2 (S. 409, Z. 29–S. 413, Z. 8). – Mairhofer Nr. 250. (74vb) Graesse Nr. 96 (90), 2 (S. 413, Z. 10–21). – Mairhofer Nr. 251. (75ra) Graesse Nr. 96 (90), 2 (S. 413, Z. 21–S. 415, Z. 2). – Mairhofer Nr. 252. (75vb) Graesse Nr. 96 (90), 2 (S. 415, Z. 3–8). – Mairhofer Nr. 253. (75vb) Tit.: De quodam milite occiso sibique devoto (Graesse Nr. 96 [90], 4. – Mairhofer Nr. 254). (76ra) Graesse Nr. 96 (90), 7. – Mairhofer Nr. 255. (76ra) Graesse Nr. 96 (90), 8. – Mairhofer Nr. 256. (76ra) Graesse Nr. 96 (90), 9. – Mairhofer Cod. 308 241 Nr. 257. (76rb) Graesse Nr. 96 (90), 5. – Mairhofer Nr. 258. (76rb) Graesse Nr. 96 (90), 10. – Mairhofer Nr. 259. (76va) ‹Conradus Eberbacensis›: Exordium magnum Cisterciense, Dist. 4, cap. 2,1–30 (PL 185, 1096D–1097B. – Mairhofer Nr. 260). (76vb–78vb) Catharina: (76vb) Tit.: De virtutibus beatissime Katherine virginis et martiris (Graes se Nr. 212 [209]. – Mairhofer Nr. 261). (78rb) Graesse 172 (167) (S. 795, Z. 5–9). – Mairhofer Nr. 262. (78va) Graesse 172 (167) (S. 795, Z. 10–20). – Mairhofer Nr. 263. (78vb) Mairhofer Nr. 264. (80rb) Mairhofer Nr. 265. (80vb) Mairhofer Nr. 266. (81rb–va) Agnes: (81rb) Graesse Nr. 24, 1 (S. 116, Z. 2–8). – Mairhofer Nr. 267. (81rb) Graesse Nr. 24, 3. – Mairhofer Nr. 268. (81va) Tit.: De ymagine beate Agnetis (Graesse Nr. 24, 4 [S. 116, Z. 16–26]. – Mairhofer Nr. 269). (81va–82ra) Agatha: (81va) Tit.: De virtutibus beate Agathe (Graesse Nr. 39 [S. 173, Z. 4–14]. – Mairhofer Nr. 270). (81vb) Graesse Nr. 39 (S. 173, Z. 18–25). – Mairhofer Nr. 271. (82ra–va) De undecim milibus virginum: (82ra) Graesse Nr. 158 (153), 2. – Mairhofer Nr. 272. (82rb) Graesse Nr. 158 (153), 3. – Mairhofer Nr. 273. (82va–83ra) Margarita: Graesse Nr. 151 (146). – Mairhofer Nr. 274. (83ra–84va) Thais meretrix: (83ra) Graesse Nr. 152 (147) (S. 677, Z. 25–S. 679, Z. 24). – Mairhofer Nr. 275. (84rb) Graesse Nr. 152 (147) (S. 679, Z. 24–31). – Mairhofer Nr. 276. Expl. erweitert: … Illa autem confusa recessit et postea penitencia ducta tandem conversa est ad domi num. (84va–85va) Prothus et Iacinthus: Graesse Nr. 136 (129) (S. 602, Z. 14, Z. 17–S. 604, Z. 31). – Mairhofer Nr. 277. (85vb–92ra) Elisabeth: (85vb) Mairhofer Nr. 278. (91ra) Mairhofer Nr. 279. (92ra) Schlussschrift: Explicit distinccio VIIa. 1.5 (92ra–109va) Dist. 8: Miracula de defunctis. (92ra–vb) Register mit Kapitelzahlen. (92vb–93rb) De omnibus sanctis. Tit.: De festo omnium sanctorum et memoria mortuorum qualiter ortum habuerunt (Graesse Nr. 162 [157] [S. 727, Z. 12–S. 728, Z. 4]). Inc. gegenüber Ed. leicht abweichend, übereinstimmend mit Mairhofer Nr. 280. (93rb–97rb) De commemoratione animarum: (93rb) Graesse Nr. 163 (158) (S. 731, Z. 4–24). – Mairhofer Nr. 281. (93vb) Graesse Nr. 163 (158) (S. 731, Z. 26–S. 732, Z. 9). – Mairhofer Nr. 282. (94ra) Graesse Nr. 163 (158) (S. 732, Z. 13–22). – Mairhofer Nr. 283. (94rb) Graesse Nr. 163 (158) (S. 732, Z. 34–S. 733, Z.11). – Mairhofer Nr. 284. (94va) Tit.: De mortuis qui vivum defenderunt (Graesse Nr. 163 [158] [S. 733, Z. 20–25]. – Mairhofer Nr. 285). (94vb) Graesse Nr. 163 (158) (S. 733, Z. 11–18). – Mairhofer Nr. 286. (94vb) Graesse Nr. 163 (158) (S. 733, Z. 30–S. 734, Z. 19). – Mairhofer Nr. 287. (95rb) Mairhofer Nr. 288. (95vb) Graesse Nr. 163 (158) (S. 734, Z. 33–S. 735, Z. 3). – Mairhofer Nr. 289. (96ra) Graesse Nr. 163 (158) (S. 735, Z. 18–26). – Mairhofer Nr. 290. (96rb) Graesse Nr. 163 (158) (S. 735, Z. 32–S. 736, Z. 8). – Mairhofer Nr. 291. (96va) Graesse Nr. 163 (158) (S. 736, Z. 9–13). – Mairhofer Nr. 292. (96vb) Graesse Nr. 163 (158) (S. 738, Z. 34–S.739, Z. 13). – Mairhofer Nr. 293. (97ra) Graesse Nr. 163 (158) (S. 739, Z. 19–29). – Mairhofer Nr. 294. (97rb) Ed.: J. Klapper, Exempla aus Handschriften des Mittelalters (Sammlung mittellateinischer Texte II). Heidelberg 1911, Nr. 26, 27. – Mairhofer Nr. 295. (97va) Mairhofer Nr. 296. (97va) Mairhofer Nr. 297. (97vb) Mairhofer Nr. 298. (98ra) Tit.: De duobus fratribus sacerdotibus horribile (Mairhofer Nr. 299). (98vb) ‹Innocentius III.›: De con temptu mundi, Lib. 3, cap. 7 (PL 217, 740A. – Mairhofer Nr. 300). (99ra) Mairhofer Nr. 301. (99vb–101rb) ‹Honorius Augustodunensis›: Speculum ecclesiae. Sermo generalis: (99vb) Ad milites (PL 172, 865A–C. – Mairhofer Nr. 302). (100ra) Ad pauperes (PL 172, 864C–865A. – Mairhofer Nr. 303). (100rb) Ad agricolas (PL 172, 866B–867B. – Mairhofer Nr. 304). (101ra) Ad coniugatos (PL 172, 870A–B. – Mairhofer Nr. 305). 242 Cod. 308 (101rb–vb) ‹Herbertus Claraevallensis›: De miraculis = ‹Conradus Eberbacensis›: Exordium magnum Cisterciense: (101rb) Lib. 2, cap. 33 (PL 185, 1344B–D) = Dist. 4, cap. 35, 1–48 (PL 185, 1123B–1124A). – Mairhofer Nr. 306. (101va) Lib. 2, cap. 34 (PL 185, 1344D–1345B) = Dist. 5, cap. 2, 41–64 (PL 185, 1126D–1127B). – Mairhofer Nr. 307. (101vb) Lib. 2, cap. 35 (PL 185, 1345C–D) = Dist. 5, cap. 3, 11–44 (PL 185, 1127C–1128B). – Mairhofer Nr. 308. (102ra–103va) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum: (102ra) Lib. 2, cap. 204, 1–6 (PL 179, 1188B–1189C. – Mairhofer Nr. 309). (103ra) Lib. 2, cap. 204, 7 (PL 179, 1189D–1190A. – Mairhofer Nr. 310). (103ra) Lib. 3, cap. 293, 1–2 (PL 179, 1262A–C. – Mair hofer Nr. 311). (103va) Mairhofer Nr. 312. (104ra) Mairhofer Nr. 313. (105ra) Mairhofer Nr. 314. (105va) Mairhofer Nr. 315. (106va) Tit.: De clerico qui nepotem suum de monasterio extraxit (Mairhofer Nr. 316). (107rb) Mairhofer Nr. 317. (107va–109ra) ‹Gregorius Magnus›: Dialogi, Lib. 4: (107va) Cap. 33 (PL 77, 372C–373A. – Mairhofer Nr. 318). (107vb) Cap. 40 (PL 77, 392C–393A. – Mairhofer Nr. 319). (108ra) Cap. 40 (PL 77, 393B–C. – Mairhofer Nr. 320). (108rb) Cap. 54 (PL 77, 413B–C. – Mairhofer Nr. 321). (108va) Cap. 55 (PL 77, 416A–B. – Mairhofer Nr. 322). (108vb) Cap. 56 (PL 77, 416B–C. – Mairhofer Nr. 323). (108vb) Cap. 53 (PL 77, 412C–413A. – Mairhofer Nr. 324). (109ra) Graesse Nr. 180 (175) (S. 816, Z. 12–33). – Mairhofer Nr. 325. (109va) Schlussschrift: Explicit distinccio octava. 1.6 (109va–131ra) Dist. 9: Miracula de sacramentis ecclesiae. (109va) Tit.: Incipit distinccio nona que agit per ordinem de sacramentis ecclesie et continet miracula LIIIIor. (109va–110rb) Register mit Kapitelzahlen. (110rb–111va) Sacramentum baptismi: (110rb) Graesse Nr. 52, 2. – Mairhofer Nr. 326. (111rb) Graesse Nr. 117 (112) (S. 491, Z. 1–7); anschließend Ergänzung zur hl. Odilia, vgl. Graesse Nr. 190 (189). – Mairhofer Nr. 327. (111va) Graesse Nr. 181 (176), 1 (S. 824, Z. 24–28). – Mairhofer Nr. 328. (111va) Graesse Nr. 181 (176) (S. 834, Z. 3–10). – Mairhofer Nr. 329. (111va) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 32 (PL 185, 1377B–1378C. – Mairhofer Nr. 330). (112va) Mairhofer Nr. 331. (113ra) Sacramentum confirmationis: Mairhofer Nr. 332. (113va–114rb) Unctio infirmorum: (113va) Mairhofer Nr. 333. (114ra) Mairhofer Nr. 334. (114rb–117ra) Sacramentum eucharistiae: (114rb–116ra) ‹Paschasius Radbertus›: De corpore et sanguine dni, cap. 14: (114rb) PL 120, 1317B–C. – Mairhofer Nr. 335. (114rb) PL 120, 1318B–1319C. – Mairhofer Nr. 336. (115ra) PL 120, 1319D–1320D. – Mairhofer Nr. 337. (115va) Mairhofer Nr. 338. (115vb) Mairhofer Nr. 339. (116ra) ‹Herbertus Claraevallensis›: De miraculis, Lib. 1, cap. 20 (PL 185, 1296D–1297D. – Mairhofer Nr. 340). (116va) Tit.: De quodam sacerdote inmundo et corpore Christi indigno terribile signum (Mairhofer Nr. 341). (116vb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 28–29 (PL 185, 1373D–1374D. – Mairhofer Nr. 342). (117ra) Mairhofer Nr. 343. (117ra–126rb) Sacramentum paenitentiae et reconciliationis: (117ra) Mairhofer Nr. 344. (117vb) Mairhofer Nr. 345. (118ra) ‹Grimlaicus›: Regula solitariorum, cap. 36 (PL 103, 625D–626A. – Mairhofer Nr. 346). (118rb) Mairhofer Nr. 347. (118va) Mairhofer Nr. 348. (119ra) Mairhofer Nr. 349. (119va) Mairhofer Nr. 350. (120ra) Mairhofer Nr. 351. (120va) Mairhofer Nr. 352. (121va) ‹Sigbertus Gemblacensis›: Chronica 823 (PL 160, 157C. – Mairhofer Nr. 353). (121va) ‹Odo de Cheriton›, Parabolae 188 (Ed. Hervieux IV 339. – Mairhofer Nr. 354). (121vb) Mair hofer Nr. 355. (122vb) Mairhofer Nr. 356. (122vb) Graesse Nr. 181 (176), 4 (S. 834, Z. 17–21). – Mairhofer Nr. 357. (123ra) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 175 (PL 179, 1146D–1147A. – Mairhofer Nr. 358). (123vb) Tit.: De quodam dyacono Cod. 308 243 qui tonsuram deposuit (Mairhofer Nr. 359). (123vb) ‹Herbertus Claraevallensis›: De m iraculis, Lib. 3, cap. 13 (PL 185, 1365C–D. – Mairhofer Nr. 360). (123vb) Mairhofer Nr. 361. (124rb) Mairhofer Nr. 362. (124vb) Mairhofer Nr. 363. (125rb) Mairhofer Nr. 364. (125va) ‹Sulpicius Severus›: Dialogi, Lib. 1, cap. 14 (PL 20, 192D–193B. – Mairhofer Nr. 365). (126ra) Tit.: De latrone qui veniam meruit propter IIIIor candelas (Mairhofer Nr. 366). (126rb) Mairhofer Nr. 367. (126vb) Tit.: De obsesso qui monachum non confessum confudit (Mairhofer Nr. 368). (126vb) ‹Caesarius Heisterbacensis›: Dialogus miraculorum, Dist. 3, cap. 6. Tit.: Item aliud de eodem (Ed. Strange I, 116–120. – Mairhofer Nr. 369). (127rb) Tit.: Quod confessio non tantum anime sed eciam corpori prodest (Mairhofer Nr. 370). (127va) Mairhofer Nr. 371. (127va) Mairhofer Nr. 372. (127vb) Mairhofer Nr. 373. (127vb) Mairhofer Nr. 374. Expl.: … sine confessione et viatico exspiravit. (128ra) Mairhofer Nr. 375. (128rb–131ra) Sacramentum matrimonii: (128rb) ‹Herbertus Claraevallensis›: De miraculis, Lib. 3, cap. 34 (PL 185, 1379C–1381B. – Mairhofer Nr. 376). (129rb) Mairhofer Nr. 377. (129va) Mairhofer Nr. 378. Expl.: … angelorum consorcia in celestibus meruerunt. (130ra) Mairhofer Nr. 379. (131ra) Schlussschrift: Explicit distinccio IXa. 1.7 (131ra–187vb) Dist. 10: Miracula de factis et dictis theologorum. (131ra) Tit.: Incipit distinccio Xa que agit de diversis videlicet factis et dictis theologorum et continent (!) sub compendio capitula CCa et XXa valde utilia necnon et delectabilia (Mairhofer Bl. 118vb). (131ra–133va) Register mit Kapitelzahlen. (133va) Ed.: R. Lotti, Contributi su Domenico Cavalca (c. 1270–1342), Diss. Amsterdam 1987, 231–232. – Mairhofer Nr. 390. (133vb) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 174 (PL 179, 1146A–C. – Mairhofer Nr. 391). (134rb) Mairhofer Nr. 392. (134va) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 207 (PL 179, 1192A–B. – Mairhofer Nr. 393). (134va) Mairhofer Nr. 394. (135va) Mairhofer Nr. 395. (136rb) ‹Guilelmus Malmesburiensis›: Gesta regum Anglorum, Lib. 2, cap. 192 (PL 179, 1172A–B. – Mairhofer Nr. 396). (136vb–138rb) ‹Sulpicius Severus›: Dialogi, Lib. 1: (136vb) Cap. 15 (PL 20, 193D–194B. – Mairhofer Nr. 397). (137ra) Cap. 17 (PL 20, 194D–195A. – Mairhofer Nr. 398). (137rb) Cap. 18 (PL 20, 195C. – Mairhofer Nr. 399). (137vb) Cap. 19 (PL 20, 195D–196B. – Mairhofer Nr. 400). (138ra) Cap. 20 (PL 20, 196B–197A. – Mairhofer Nr. 401). (138rb) ‹Wernerus de Ellerbach›: Deflorationes ss. patrum, Lib. 1, Sermo in rogationibus (PL 157, 968B–C. – Mairhofer Nr. 402). (138va–139va) ‹Honorius Augustodunensis›: Speculum ecclesiae: (138va) Dnca 1. post pent. (PL 172, 1042B–D. – Mairhofer Nr. 403). (139ra) Dnca 20. post pent. (PL 172, 1066D–1068A. – Mairhofer Nr. 404). (139va–141ra) ‹Gregorius Magnus›: XL Homiliae in Evangelia: (139va) Lib. 1 (PL 76, 1133C–1134B. – Mairhofer Nr. 405). (140ra) Lib. 2 (PL 76, 1300B–D. – Mairhofer Nr. 406). (140rb) Lib. 1 (PL 76, 1158B–1159A. – Mairhofer Nr. 407). (141ra–144va) Ioseph: (141ra) Graesse Nr. 67 (63), 2 (S. 298, Z.15–S. 299, Z. 13). – Mairhofer Nr. 408. (141vb) Graesse Nr. 67 (63), 2 (S. 299, Z. 15–S. 300, Z. 16). – Mairhofer Nr. 409. (142rb) Graesse Nr. 67 (63), 2 (S. 300, Z. 16–301, Z. 16). – Mairhofer Nr. 410. (143ra) Graesse Nr. 67 (63), 2 (S. 301, Z. 16–36). – Mairhofer Nr. 411. (143rb) Graesse Nr. 67 (63), 2 (S. 301, Z. 37–S. 302, Z. 11). – Mairhofer Nr. 412. (143vb) Graesse Nr. 67 (63), 2 (S. 302, Z. 11–32). – Mairhofer Nr. 413. (144ra) Graesse Nr. 67 (63), 2 (S. 302, Z. 32–S. 303, Z. 9). – Mairhofer Nr. 414. (144rb) Graesse Nr. 67 (63), 2 (S. 303, Z. 9–15). – Mairhofer Nr. 415. (144va) Iudas Iscariot: Graesse Nr. 45 (S. 184, Z. 1–S. 186, Z. 13). – Mairhofer Nr. 416. 244 Cod. 308 (145vb–147vb) Pilatus: (145vb) Graesse Nr. 53 (51) (S. 231, Z. 21–S. 232, Z. 24). – Mairhofer Nr. 417. (146va) Graesse Nr. 53 (51) (S. 232, Z. 30–233, Z. 26). – Mairhofer Nr. 418. (147ra) Graesse Nr. 53 (51) (S. 233, Z. 27–S. 234, Z. 26). – Mairhofer Nr. 419. (147vb–148vb) Nero: (147vb) Graesse Nr. 89 (84), 3 (S. 376, Z. 5–22). – Mairhofer Nr. 420. (148ra) Graesse Nr. 89 (84), 3 (S. 376, Z. 25–377, Z. 14). – Mairhofer Nr. 421. (148va) Graesse Nr. 89 (84), 3 (S. 377, Z. 15–29). – Mairhofer Nr. 422. (148vb–150ra) Iulianus Apostata: (148vb) Graesse Nr. 30, 5 (S. 143, Z. 35–S. 144, Z. 11). – Mairhofer Nr. 423. (149ra) Graesse Nr. 30, 5 (S. 144, Z. 12–23). – Mairhofer Nr. 424. (149rb) Graesse Nr. 125 (120), 1 (S. 571, Z. 3–8). – Mairhofer Nr. 425. (149rb) Graesse Nr. 125 (120), 1 (S. 571, Z. 8–14). – Mairhofer Nr. 426. (149va) Graesse Nr. 125 (120), 1 (S. 571, Z. 14–21). – Mairhofer Nr. 427. (149va) Graesse Nr. 125 (120), 1 (S. 571, Z. 21–28). – Mairhofer Nr. 428. (149vb) Graesse Nr. 30, 5 (S. 144, Z. 34–35). – Mairhofer Nr. 429. Inc.: In odium quoque Chris tianorum Iulianus templum … (150ra–va) Basilius: (150ra) Graesse Nr. 30, 5 (S. 144, Z. 35–S. 145, Z. 6). – Mairhofer Nr. 430. (150ra) Graesse Nr. 30, 5 (S. 145, Z. 6–30). – Mairhofer Nr. 431. (150va) Graesse Nr. 181 (176), 4 (S. 838, Z. 9–15). – Mairhofer Nr. 432. (150va) Graesse Nr. 181 (176), 4 (S. 837, Z. 31–38). – Mairhofer Nr. 433. (150vb) Graesse Nr. 181 (176), 4 (S. 837, Z. 13–21). – Mairhofer Nr. 434. (151ra) Graesse Nr. 181 (176), 4 (S. 836, Z. 37–S. 837, Z. 10). – Mairhofer Nr. 435. (151rb) Graesse Nr. 181 (176), 4 (S. 839, Z. 1–12). – Mairhofer Nr. 436. (151va) Mairhofer Nr. 437. (152ra) Mairhofer Nr. 438. (152rb) Graesse Nr. 181 (176), 4 (S. 840, Z. 3–841, Z. 5). – Mairhofer Nr. 439. (153rb) Mairhofer Nr. 440. (153vb) Graesse Nr. 136 (129) (S. 603, Z. 4–9). – Mairhofer Nr. 441. (153vb) Mairhofer Nr. 442. (154ra) Graesse Nr. 182 (177) (S. 846, Z. 39–S. 847, Z. 14). – Mairhofer Nr. 443. (154rb) Graesse Nr. 182 (177) (S. 854, Z. 9–17). – Mairhofer Nr. 444. (154rb) Mairhofer Nr. 445. (154va) Mairhofer Nr. 446. (154vb) 3. Esr 3,3–4,47. – Mairhofer Nr. 447. (156rb) Tit.: De XVm signis ante diem iudicii (Graesse Nr. 1 [S. 6, Z. 35–7, Z. 24]). – Expl.: … resurgent omnes et tunc apparebit filius hominis in aere. (156va–162rb) ‹Odo de Cheriton›: Parabolae: (156va) Nr. 6 (Ed. Hervieux IV 267. – Mairhofer Nr. 448). (156vb) Nr. 8 (Ed. Hervieux IV 268. – Mairhofer Nr. 449). (157ra) Mairhofer Nr. 450. (157ra) Nr. 12 und 13 (Ed. Hervieux IV 271. – Mairhofer Nr. 451). (157rb) Nr. 26 (Ed. Hervieux IV 275. – Mairhofer Nr. 452). Expl.: … cadaver illius cuius fetorem abhorrens. (157rb) Nr. 42 (Ed. Hervieux IV 281. – Mairhofer Nr. 453). (157va) Nr. 45 (Ed. Hervieux IV 282. – Mairhofer Nr. 454). (157va) Nr. 46 (Ed. Hervieux IV 282. – Mairhofer Nr. 455). Expl.: … da exemplum. (157vb) Nr. 47 (Ed. Hervieux IV 283. – Mairhofer Nr. 456). (158ra) Nr. 52 (Ed. Hervieux IV 285. – Mairhofer Nr. 457). (158ra) Mairhofer Nr. 458. (158rb) Nr. 74 (Ed. Hervieux IV 294. – Mairhofer Nr. 459). Expl.: … comedo. Hoc ordo. (158va) Nr. 84 (Ed. Hervieux IV 298. – Mair hofer Nr. 460). (158va) Nr. 86 (Ed. Hervieux IV 298–299. – Mairhofer Nr. 461). (158vb) Nr. 88 (Ed. Hervieux IV 299. – Mairhofer Nr. 462). (158vb) Mairhofer Nr. 463. (159ra) Nr. 93 (Ed. Hervieux IV 301. – Mairhofer Nr. 464). (159ra) Mairhofer Nr. 465. (159rb) Nr. 102 (Ed. Hervieux IV 304. – Mairhofer Nr. 466). (159va) Nr. 105 (Ed. Hervieux IV 305. – Mairhofer Nr. 467). (160vb) Nr. 116 und 117 (Ed. Hervieux IV 309–310. – Mairhofer Nr. 468). (160ra) Nr. 128. Tit.: Quod verus religiosus laudes et vituperia vilipendit (Ed. Hervieux IV 313–314). (160ra) Vgl. Nr. 123 (Ed. Hervieux IV 312. – Mairhofer Nr. 469). (160ra) Nr. 132 (Ed. Hervieux IV 315– 316. – Mairhofer Nr. 470). (160rb) Nr. 131 (Ed. Hervieux IV 315. – Mairhofer Nr. 471). (160va) Nr. 136 (Ed. Hervieux IV 217. – Mairhofer Nr. 472). (161ra) Nr. 133 (Ed. Hervieux IV 316. – Mairhofer Nr. 473). (161ra) Nr. 143 (Ed. Hervieux IV 320–321. – Mairhofer Nr. 474). (161rb) Nr. 144. Tit.: Qui hic contempnitur in celo consolatur (Ed. Hervieux IV 321. – Mairhofer Nr. 475). (161va) Nr. 153. Tit.: De cupiditate, luxuria et superbia (Ed. Hervieux IV 325. – Mairhofer Nr. 476). (161va) Nr. 163 (Ed. Hervieux IV 328. – Mairhofer Nr. 477). (161vb) Nr. 164 (Ed. Hervieux IV 328–329. – Mairhofer Nr. 478). (161vb) Nr. 157 (Ed. Hervieux IV Cod. 308 245 329–330. – Mairhofer Nr. 479). (162ra) Nr. 171. Tit.: De utilitate infirmitatis (Ed. Hervieux IV 332. – Mairhofer Nr. 480). (162rb) Mairhofer Nr. 481. (162rb) Mairhofer Nr. 482. (162va) Mairhofer Nr. 483. (162va) Mairhofer Nr. 484. (162vb) Graesse Nr. 180 (175) (S. 815, Z. 24–S. 816, Z. 9). – Mairhofer Nr. 485. (163ra) Mairhofer Nr. 486. (163rb) Mairhofer Nr. 487. (163rb–164vb) ‹Grimlaicus›: Regula solitariorum: (163rb) Cap. 44 (PL 103, 635B–C. – Mair hofer Nr. 488). (163va) Cap. 56 (PL 103, 847D–648A. – Mairhofer Nr. 489). (163vb) Cap. 54 (PL 103, 615D. – Mairhofer Nr. 490). Expl.: … perimunt et vanam gloriam ostendunt. (164ra) Cap. 57 (PL 103, 648D. – Mairhofer Nr. 491). (164ra) Cap. 39 (PL 103, 630B. – Mairhofer Nr. 492). (164rb) Mairhofer Nr. 493. (164va) Mairhofer Nr. 494. (164va) Vitae patrum, Lib. 5, Verba seniorum, libell. 16, cap. 19 (PL 73, 973B–C) = ‹Grim laicus›: Regula solitariorum, cap. 59 (PL 103, 654A–B. – Mairhofer Nr. 495). (164vb) Mairhofer Nr. 496. (164vb) Mairhofer Nr. 497. Expl.: … et obiit in conversacione bona. (165ra) Tit.: Quod efficaciora sunt exempla (Mairhofer Nr. 498). (165rb) Mairhofer Nr. 499. (165va) Mairhofer Nr. 500. (165va) Mairhofer Nr. 501. (165vb) Mairhofer Nr. 502. (166ra) Mairhofer Nr. 503. (166rb) ‹Gregorius Magnus›: Dialogi, Lib. 4, cap. 11 (PL 77, 336B–337A. – Mairhofer Nr. 504). (166va) Mairhofer Nr. 505. (166va) Vitae patrum, Lib. 5, Verba seniorum, libell. 3, cap. 23 (PL 73, 864A–B. – Mairhofer Nr. 506). (166va) Mairhofer Nr. 507. (166vb) Mairhofer Nr. 508. (166vb) Tit.: Quod nichil nobis manet nisi quod pauperibus damus (Mair hofer Nr. 509). (167ra) Mairhofer Nr. 510. (167rb) Mairhofer Nr. 511. (167rb) Mairhofer Nr. 512. (167va) Vitae patrum, Lib. 5, Verba seniorum, libell. 6, cap. 18 (PL 73, 891C–D. – Mairhofer Nr. 513). (167vb) Mairhofer Nr. 514. (167vb) Mairhofer Nr. 515. (168ra) Vitae patrum, Lib. 5, Verba seniorum, libell. 6, cap. 21 (PL 73, 892B–C. – Mairhofer Nr. 516). (168rb) Mairhofer Nr. 517. (168va) Mairhofer Nr. 518. (168vb) Vitae patrum, Lib. 5, Verba seniorum, libell. 3, cap. 14 (PL 73, 862B–C. – Mairhofer Nr. 519). (168vb) Mairhofer Nr. 520. (169ra) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 55 (PL 73, 768C–D) = ‹Grimlaicus›: Regula solitariorum, cap. 39 (PL 103, 630C–D). – Mairhofer Nr. 521. (169rb) ‹Hieronymus›: Epistola 107 (ad Laetam) (PL 22, 872. – Mairhofer Nr. 522). (169rb) Mairhofer Nr. 523. (169va) Mairhofer Nr. 524. (169vb) Mairhofer Nr. 525. (169vb) Mairhofer Nr. 526. (170rb) Mairhofer Nr. 527. (170rb) Mairhofer Nr. 528. (170va) Mairhofer Nr. 529. (170vb) Vgl. Vitae patrum, Lib. 5, Verba seniorum, libell. 8, cap. 24 (PL 73, 909C. – Mairhofer Nr. 530). (170vb) Mairhofer Nr. 531. (171ra) Mairhofer Nr. 532. (171rb) Mairhofer Nr. 533. (171rb) ‹Petrus Cantor›: Verbum abbreviatum, cap. 86 (PL 205, 256D–257A. – Mairhofer Nr. 534). (171va) ‹Nicolaus Pergamenus›: Dialogus creaturarum, dial. 75, 23–36 (Ed.: Th. Graesse, Die beiden ältesten latei nischen Fabelbücher des Mittelalters des Bischofs Cyrillus „Speculum sapientiae“ und des Nico laus Pergamenus „Dialogus creaturarum“. Stuttgart 1880, 219. – Mairhofer Nr. 535). (171va) Mairhofer Nr. 536. (171vb) Mairhofer Nr. 537. (172ra) Mairhofer Nr. 538. (172rb) Mairhofer Nr. 539. (172va) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 118 (PL 73, 782C–D. – Mairhofer Nr. 540). (172vb) Mairhofer Nr. 541. (172vb) Mairhofer Nr. 542. (173ra) ‹Gregorius Magnus›: Dialogi, Lib. 4, cap. 19 (PL 77, 352A–B. – Mairhofer Nr. 543). (173ra) Mairhofer Nr. 544. (173va) Mairhofer Nr. 545. (173va) ‹Rufinus Aquileiensis›: Historia monachorum, cap. 1 (PL 21, 394C. – Mairhofer Nr. 546). (173vb) Mairhofer Nr. 547. (173vb) Mairhofer Nr. 548. (174ra) Mairhofer Nr. 549. (174ra) Mairhofer Nr. 550. (174rb) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 97 (PL 73, 778A–C) und Vitae patrum, Lib. 6, Verba seniorum, libell. 3, cap. 17 (PL 73, 1013D–1014B. – Mairhofer Nr. 551). (174va) Mairhofer Nr. 552. (174va–175ra) ‹Innocentius III.›: De contemptu mundi, Lib. 2: (174va) Cap. 36 (PL 217, 731D–732A. – Mairhofer Nr. 553). (174vb) Cap. 39 (PL 217, 733B. – Mairhofer Nr. 554). (175ra) Cap. 20 (PL 217, 725A. – Mairhofer Nr. 555). (175ra) Mairhofer Nr. 556. 246 Cod. 308 (175rb–176va) Vitae patrum, Lib. 5, Verba seniorum: (175rb) Libell. 14, cap. 52 (PL 73, 963B– C. – Mairhofer Nr. 557). (175va) Mairhofer Nr. 558. (175va) Libell. 14, cap. 72 (PL 73, 966A– B. – Mairhofer Nr. 559). (176ra) Libell. 5, cap. 34 (PL 73, 882D–883A. – Mairhofer Nr. 560). (176rb) Mairhofer Nr. 561. (176rb) Libell. 3, cap. 2 (PL 73, 860C–D. – Mairhofer Nr. 562). (176va) Libell. 17, cap. 18 (PL 73, 976B. – Mairhofer Nr. 563). (176va) Mairhofer Nr. 564. (176vb) Mairhofer Nr. 565. (177ra) ‹Petrus Cantor›: Verbum ab breviatum, cap. 98 (PL 205, 280A–B. – Mairhofer Nr. 566). (177ra) Mairhofer Nr. 567. (177rb) ‹Gregorius Magnus›: Dialogi, Lib. 1, cap. 9 (PL 77, 197B–C. – Mairhofer Nr. 568). (177va) Mairhofer Nr. 569. (177va) ‹Rufinus Aquileiensis›: Historia monachorum, cap. 31 (PL 21, 458B–C. – Mairhofer Nr. 570). (177vb) Mairhofer Nr. 571. (178ra–182ra) ‹Ps.-Rufinus Aquileiensis›: Vitae patrum: (178ra) Lib. 3, cap. 28 (PL 73, 756B–C) und Lib. 6, Verba seniorum, libell. 2, cap. 17 (PL 73, 1004A. – Mairhofer Nr. 572). (178ra) Lib. 3, cap. 13 (PL 73, 745B–746B. – Mairhofer Nr. 573). (178rb) Mairhofer Nr. 574. (178va) Mairhofer Nr. 575. (178va) Mairhofer Nr. 576. (178vb) Mairhofer Nr. 577. (178vb) Mairhofer Nr. 578. (179vb) Mairhofer Nr. 579. (180va) Mairhofer Nr. 580. (181rb) Lib. 3, cap. 84 (PL 73, 775B–C. – Mairhofer Nr. 581). (181vb) ‹Sulpicius Severus, Iohannes Cassianus›: Vitae patrum, Lib. 4, Excerpta, cap. 43 (PL 73, 841A–C. – Mairhofer Nr. 582). (182ra–183vb) ‹Rufinus Aquileiensis›: Historia monachorum: (182ra) Cap. 1 (PL 21, 399A–400A. – Mairhofer Nr. 583). (182vb) Cap. 6 (PL 21, 401C–403C. – Mairhofer Nr. 584). (183vb–187va) Vitae patrum: (183vb) ‹Sulpicius Severus, Iohannes Cassianus›: Lib. 4, E xcerpta, cap. 52 (PL 73, 846B–D. – Mairhofer Nr. 585). (184ra) Lib. 5, Verba seniorum, libell. 4, cap. 27 (PL 73, 880C–D. – Mairhofer Nr. 586). (184va) Lib. 5, Verba seniorum, libell. 5, cap. 37 (PL 73, 883D–884B. – Mairhofer Nr. 587). (185ra) Lib. 5, Verba seniorum, libell. 5, cap. 30 (PL 73, 881C. – Mairhofer Nr. 588). (185ra) Lib. 5, Verba seniorum, libell. 7, cap. 41 (PL 73, 903A. – Mairhofer Nr. 589). (185rb) Lib. 5, Verba seniorum, libell. 7, cap. 43 (PL 73, 903B–904A. – Mairhofer Nr. 590). (185vb) Lib. 5, Verba seniorum, libell. 10, cap. 10 (PL 73, 913C–D. – Mairhofer Nr. 591). (186ra) Lib. 5, Verba seniorum, libell. 14, cap. 18 (PL 73, 952A) = ‹Grimlaicus›: Regula solitariorum, cap. 58 (PL 103, 649D–650A). – Mairhofer Nr. 592. (186rb) Lib. 6, Verba seniorum, libell. 1, cap. 12 (PL 73, 994C–995A. – Mairhofer Nr. 593). (186vb) Lib. 6, Verba seniorum, libell. 1, cap. 13 (PL 73, 995A–B. – Mairhofer Nr. 594). (187ra) ‹Ps.- Rufinus Aquileiensis›: Vitae patrum, Lib. 3, Verba seniorum, cap. 54 (PL 73, 768B–C. – Mair hofer Nr. 595). (187ra) Lib. 3, Verba seniorum, cap. 55 (PL 73, 768D. – Mairhofer Nr. 596). (187va) ‹Godefridus Altissiodorensis›: Declamationes ex s. Bernardi sermonibus, cap. 25 (PL 184, 454C–455A. – Mairhofer Nr. 597). (187vb) Schlussschrift: Hic explicit liber qui dicitur Lacteus liquor. Post hec incipiunt alia exem pla que non videbantur superius contineri. Lit.: Zur Verfasserfrage und Überlieferung s. Mairhofer 9–74. Zur Lacteus liquor-Sammlung am Beispiel von Heiligenkreuz, Stiftsbibl., Cod. 323 vgl. Welter 279–281. 2 (187vb–201rb) 27 weitere Exempla: Miracula de diversis. Unmittelbar an Lacteus liquor anschließend und nach der Angabe am oberen Blattrand zu Dist. 10 gehörend. (187vb) Tit.: De sancto Thoma martyre et sua domina sancta Maria. – Inc.: Sanctus Thomas Cantuariensis bone yndolis adolescens cum adhuc in scolis … – Expl.: … de medio sunt s ubtracti. (188va) Tit.: De sancto Udalrico episcopo et tribus bursis dyalogus. – Inc.: Sanctus Udalricus presul Augustensis cum quandam ecclesiam sue diocesis dedicare vellet … – Expl.: … signa ex corde ostendere. (189ra) Visio Philiberti, 1. Fassung (Ed.: L. Zatočil, Die Visio Fulberti nach einer bislang unbe achteten Brünner Handschrift aus dem Ende des 14. Jahrhunderts. Sborník prací filosofické fakul ty Brnĕnské University D. Řada literárnĕvĕdná 21 (1974) 39–47. – Walther, Initia Nr. 11894). Cod. 308 247 Tit.: Conflictus corporis et anime. – Inc.: Noctis sub silencio tempore brumali deditus … – Expl.: … et me Christi manibus totum commendavi. Lit.: N. F. Palmer, „Visio Philiberti“. Verfasserlexikon2 X 412–418. (191rb) Tit.: De quodam milite suspenso et virtute corporis Christi. – Inc.: Miles quidam aput Ffridricum imperatorem in multis est accusatus … – Expl.: … miraculum divulgantes. (191vb) Tit.: De virtute confessionis et quodam iuvene qui apostatavit (enthalten in: J. Major, Magnum speculum exemplorum. Circino aureo 1611, 44–45 [Nr. III]). (192vb) Tit.: Quod infirmitas cum paciencia mitigat penam purgatorii. – Inc.: Monachus quidam vite laudabilis sed corpore debilis fuit … – Expl.: … admodum superius est relatum. (193ra) Tit.: De quodam magno latrone et virtute penitencie. – Inc.: Legitur quod in dyocesi Tridentina quidam latro fuerat famosus … – Expl.: … virginis devotissime ieiunare. (194ra) Tit.: De sancto Bernhardo et monacho suo qui habuit multos pueros (enthalten in: J. Major, Magnum speculum exemplorum. Circino aureo 1611, 46 [Nr. VI]). (194va) Tit.: De quodam usurario qui vidit duos nigros equos et de dyabolo. – Inc.: Usurarius quidam cum crux in quibusdam partibus predicaretur cum ceteris cecus non ex devocione sed ex importuna circumstancium ammonicione … – Expl.: … et viatico est defunctus et sepultus in inferno. (195rb) Tit.: De usurario et eius corde inexplebili. – Inc.: Usurarius quidam dives nimis diversa rum … – Expl.: … postmodum aliis enarravit. (196rb) Tit.: De sancto Thoma et milite. – Inc.: Miles quidam beatum Thomam apostolum arden ter diligebat … – Expl.: … sponso valedixit. (197ra) Tit.: Miraculum de decima. – Inc.: Miles quidam in dandis decimis suis tante erat iusticie ut ei summum esset studium … – Expl.: … uberiores fructus fecit. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 285, Bl. 107r. (197rb) Tit.: De quodam converso et malo consilio dyaboli. – Inc.: Conversus quidam in quodam cenobio cum monachis loquendo … – Expl.: … et anima confundatur. (197vb) Tit.: De sancto Severino et purgatorio. – Inc.: Legitur de sancto Severino quod vivens mortuos suscitavit ymo omnem infirmitatem … – Exp.: … post hec disparuit. (198rb) Tit.: De dyabolo et globo igneo. – Inc.: Fuit quidam vir lascivus valde contigit eum … – Expl.: … possum moveri pede. (198va–200va) ‹Gregorius Magnus›: Dialogi (CPL Nr. 1713). (198va) Lib. 3, cap. 38 (PL 77, 316A–317A). Tit.: De sancto Eutico et de mundo fugiendo. – (198vb) Lib. 3, cap. 4 (PL 77, 224C–225A). Tit.: De diversis vocibus dyaboli propter super biam. – (199ra) Lib. 3, cap. 5 (PL 77, 225C–228B). Tit.: De ceco episcopo et veneno. – (199rb) Lib. 3, cap. 10 (PL 77, 236B–237A). Tit.: Quod elementa Deo et sanctis obediunt homines vero non. – (199rb) Lib. 3, cap. 11 (PL 77, 237A–B). Tit.: De virtute hospitalitatis. – (199va) Lib. 4, cap. 36 (PL 77, 381C). Tit.: De suppliciis inferni. – (199vb) Lib. 4, cap. 32 (PL 77, 372C–373A). Tit.: De cuiusdam morte subitanea. – (200ra) Lib. 3, cap. 14 (PL 77, 218B–C). Tit.: Qui de trahunt aliena et hereditabunt vermes et serpentes. – (200ra) Lib. 3, cap. 15 (PL 77, 253C–D). Tit.: De serpentibus qui propter oracio (!) mortui sunt et propter oracionem aves d eportaverunt. – (200rb) Lib. 3, cap. 16 (PL 77, 257B–D). Tit.: De dyabolo qui per formam serpentis apostatare studuit quendam sanctum. (200va) Tit.: De risione dyabolorum pro uno calcio. – Inc.: Legitur de sancto Udalrico cum vellet causa devocionis … – Expl.: … peccatum mortale evasit. (200vb) Tit.: De rustico paupere et morte. – Inc.: Fuit quidam rusticus qui cum iret … – Expl.: … maioribus sanctis in celo. (201rb) Schlussschrift: Expliciunt miracula de diversis. 248 Cod. 308 3 (201va–202ra) Mirakel. (201va–b) Tit.: Prolacio. – Inc.: Primo debemus querere Christum cum plena cordis conversio ne … – Expl.: … pereunt omnia prius facta. (201vb–202ra) Tit.: Miraculum. – Inc.: Exemplum legitur quod quidam pauper manibus labora bat … – Expl.: … gracias Deo retulit. Confidamus ergo fratres in solo Deo. Parallelüberlieferung: Schlägl, Stiftsbibl., Cod. 202, Bl. 228v. (202rb–vb) nur Rahmung und Linierung. 4 (203ra–210ra) Übersetzung vom Lat. ins Dt. einzelner Wörter des Psalteriums, der Cantica, des Pater noster, des Credo, des Te Deum, des Symbolum Athanasianum und des Ave Maria. Jeweils Anfang der einzelnen Psalmen und Gebete angeführt, anschließend ausgewählte Wörter über setzt (lat. Wörter in Schwarz, dt. Übersetzung in Rot, meist darüber, vereinzelt daneben). Tit.: Vlgarizacio (!) aliquorum vocabulorum psalterii. – Inc.: Beatus vir qui non abiit / Pestilencie (darüber:) Der Suechte … – Expl.: … Alvo. Pauch. Coadunare. zů fuegen. 5 (210ra–225va) Tractatus de decem praeceptis. (210ra) Prolog. Tit.: De incredulitate hominum prologus. – Inc.: Audi Israel Deus tuus unus est (Dt 6,4). Sunt autem septem genera hominum qui Deum non colunt sed ydola adorant. Primus genus est de quibus … – Expl.: … regnum Dei non videbunt. (211rb) Text. Inc.: Non sumas nomen Dei tui invanum. Augustinus. Nomen Dei sumit invanum qui semper turpia verba profert ex corde. Unde legitur in dyalogo Gregorii … – Expl.: … vel incendunt vel depredantur, hos neque episcopus absolvere nisi gracia apostolici etc. Traktat endet hier nach dem siebten Gebot. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 230, Bl. 155ra (bis Bl. 162rb übereinstimmend) (219ra–b) am unteren Blattrand Ergänzung von anderer Hand: Exemplum de quodam rege qui habuit duos camerarios valde invidos … et hoc fecit propter invidiam ut suo socio ambo oculi eruerentur. 6 (225va–228rb) Sentenzen zum Dies Sabbati verschiedener Auctoritates, u. a. Hieronymus, Augustinus, Gregorius, Seneca, Beda Venerabilis, Malachias, Ambrosius, Eusebius. Tit.: Auctoritates sanctorum de diversis sentenciis. – Inc.: Memento ut sanctifices diem sabbati. Per sabbatum intelligimus sollempnitates sanctorum in quibus quam plurimum homines vacant commessacionibus et ebrietatibus … – Expl.: … Bernhardus. Recole primordia tua hec pudorem addunt. Attende media illa dolorem ingerunt. Memorare novissima illa mecum incuciunt. Cogita unde veneris et erubesce ubi sis et ingemisce et contremisce quo vadis. 7 (228rb–vb) Verse zu Stationen im Leben Mariens und Jesu (vgl. Walther, Initia Nr. 6444). 1) De annunciacione versus. – Fert paranynphus ave quo mundus solvitur ave. 2) De salutacione Marie et Elizabet. – Scandit montana pia virgo stupet veterana. 3) De nativitate domini. – Sic mundo pactum solvit verbo caro factum. 4) Christus pastoribus nunciatur. – Grex cum pastore pacis letatur amore. 5) Adoratur a magys. – Dona ferunt proceres colitur quibus inclitus heres. Edocti fato dant reges munera nato. 6) De purificacione sancte Marie. – Sistitur hic domino redimendus turture bino. 7) De occisione parvulorum. – Plebs ab ymatu parit Herodis cruciatu. 8) De fuga in Egyptum. – Dum mors grassatur puero fuga tuta paratur. 9) De sessione Ihesu. – Inter doctores populi sedet et seniores. 10) De baptismo Christi. – Iordanis sacri subit hic baptisma lavatri. / Contactu mundas nobis Deus conficit undas. 11) De mutacione aque et (radiert, korr. zu in) vinum. – Hic contra morem vini gerit unda s aporem. 12) De vocacione Petri et Andree. – Hos vocat in primis Pelagi salvator ab ymis. 13) De cura leprosi. – Mundat leprosum tangens sub lege perosum. Cod. 315 249 14) De filio regali. – Regulus obnatum stat supplex febre gravatum. 15) Convertit Samaritanam apud puteum. – Fons vite lassus solus sitit essibus assus. 16) De sedacione tempestatis maris. – (am unteren Blattrand eingefügt: Pupe saporatus iubet obmutescere natus.) Imperat sic ventis Pelagi silet unda furentis. 8 (228vb) Kurztraktat über die Legende der 30 Denare, unvollständig. Tit.: Nota. Translacionem denariorum pro quibus venditus fuit Ihesus. – Inc.: Primus qui hic fuit Ninius idem fabricator fuit Thare ille dedit Abram. Abram dedit Ysmahelitis … – Expl. mut.: … et XV custodientibus sepulchrum [ (Ir–v) leer. A. P. Cod. 315 ISAIAS mit GLOSSA ORDINARIA Perg. I, 88, I Bl. 235 × 170. Entstehungsort unbestimmt, 12. Jh. B: Perg. unregelmäßig beschnitten, tw. löchrig, Nahtspuren, tw. fleckig. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 11.IV88 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Vor der ersten Lage fehlen wohl sechs Quaternionen mit Textverlust (Bl. 1–46), nach der letzten Lage ebenfalls Textverlust. Am Lagenende Kustoden, stark beschnitten. Am rechten oberen Blatt rand Foliierung 15. Jh. in arabischen Ziffern 47–92 (Zahl 88 ausgelassen), entspricht Bl. 1–45 der Foliierung 19. Jh. durch die ULB Tirol in arabischen Ziffern 1–88 (hier berücksichtigt). S: Schriftraum 160 × 135/140. Drei Spalten von unterschiedlicher Breite, durch zweifache Tintenlinien von einander abgegrenzt. Haupttext zu 13–14 Zeilen auf Tintenlinierung. Zirkelstiche. Karolingisch-gotische Minuskel wohl von einer Hand. Rand- und Interlinearglossen. Am äußeren Rand vereinzelt Notizen von anderer Hand. A: Schwarze Paragraphzeichen. Bl. 76r und 77v rote Paragraphzeichen, Auszeichnungsstriche und Kapitel angaben. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H.18. Jh. Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren ge säumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck JSAIÆ ONUS ÆGYP., im dritten Signatur schild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neu stift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II2 D 14 (?) (Tinte, radiert), II1 D 15 (Bleistift) und 315 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 315. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol IV 90, 471. (Ir–v) leer. Isaias mit Glossa ordinaria, unvollständig: Is 18,7–59,13 (Druck: GW 4282. PL 113, 1260B–1302A). (1r) am oberen Blattrand Vermerk (18. Jh.): Caput 19 Jsaie incipit hic à verbis: Onus Egypti. (1r) Inc. mut.: ]tuum montem Syon. Onus Egypti. Ecce dominus ascendet super nubem levem … – Expl. mut.: … et loquemur calumpniam et transgressionem. Concepimus et [ Zur Frage der Verfasserschaft der Glossa ordinaria s. L. Smith, The Glossa Ordinaria. The Making of a Medieval Bible Commentary. Leiden 2009. (Ir–v) leer. P. K. 250 Cod. 327 Cod. 327 GALFREDUS DE VINOSALVO Perg. I, 21, I Bl. 220 × 150. Frankreich (?), 13. Jh. B: Perg. fleckig, Haar- und Fleischseite deutlich unterscheidbar. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + (IV– 1)7 + (IV–I)13 + (IV–III)15 + (IV–I)21 + (I–1)I* (Nachsatzbl., Pap. 18 Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Urspr. wohl durchgehend Quaternionen: Bei Lage 1 fehlt Ge genbl. zu Bl. 1, bei Lage 2 fehlt ein Doppelbl. in Lagenmitte, bei Lage 3 fehlen äußerstes Doppelbl. sowie zwei Doppelbl. in Lagenmitte, bei Lage 4 fehlt ein Doppelbl. in Lagenmitte (Textverlust). Nach Bl. 21 fehlt mindestens ein Unio (Textverlust). Bl. 13v Reklamante. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 160 × 80, von tw. kaum sichtbaren Bleistiftlinien gerahmt, zu 36 Zeilen auf tw. kaum sichtbarer Bleistiftlinierung. Textualis. Am Blattrand und interlinear Kommentare und Notizen von zeitgenössischer Hand. A: Rote Auszeichnungsstriche, Durchstreichungen, Über- und Schlussschriften sowie ein- und zweizeilige ein fache Lombarden, tw. mit Punktverdickungen. Bl. 1r Initiale P (42 mm hoch) mit rot-blau kopfstempel förmig gespaltenem Buchstabenkörper und rotem Palmettenfleuronnée. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, drei einfache Bünde, von Streicheisenlinien begrenzt. Im zweiten Feld Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen II`1 D 9 S. 2 (Tinte, Bleistift), darunter ältere radierte Signatur (erhalten: II` , Tinte) sowie 327 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 327. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller VI 122. – Wilhelm I 135. – D. Schaller, J. Stohlmann, Nachträge zu Hans Walther, Initia carminum ac versuum medi aevi. Mittellateinisches Jahrbuch 2 (1971) 293–314, hier 300. – W. Schmidt, Untersuchun gen zum „Geta“ des Vitalis Blesensis. Ratingen u. a. 1975, 10. – A. Paeske, Der „Geta“ des Vitalis von Blois. Köln 1976, 44. – Steer 36. – Katalog ULB Tirol IV 137, 471. – M. C. Woods, Classroom Commentaries. Teaching the Poetria nova across Medieval and Renaissance Europe. Columbus 2010, 144–145, 299. 1 (Ir–v) leer. (1r–19v) Galfredus de Vinosalvo: Poetria nova, unvollständig (Druck: P. Leyser, Historia poe tarum et poematum medii aevi. Halae 1721, 862–978. E. Faral, Les arts poetiques du XIIe et du XIIIe siècle [Bibliothèque de l’École des Hautes Études 238]. Paris 1924, 197–262). (1r) Tit.: Incipit poetria magistri Ganfredi. (1r–7v) Inc.: Papa stupor mundi si dixero papa nocenti … – Expl. mut. (V. 478): … puduit minus in peregrinis [ V. 515–526 bereits urspr. ausgelassen, Nachtrag der Verse und Hinweis vom selben Schreiber auf Bl. 19v. (8r–10v) Inc. mut. (V. 563): ] preformet capitis (!) nature … – Expl. mut. (V. 775): … labra flamme [ (11r–13v) Inc. mut. (V. 915): ] consiliis cinctus odiis … – Expl. mut. (V. 1130): … virtus optima rerum [ (14r–v) Inc. mut. (V. 1276): ] In serie dicta … – Expl. mut. (V. 1347): … mortale remordeat, istud [ (15r–v) Inc. mut. (V. 1498): ] Qui cecidit seseque potens … – Expl. mut. (V. 1568): … placuerunt federa pacis [ (16r–18v) Inc.: mut.: (V. 1709): ] Vocum que flecti nequeunt … – Expl. mut. (V. 1923): … per horret hyatus [ Cod. 353 251 (19r–v) Inc. mut. (V. 2066): ] Iam mare transcurri … – Expl.: … de iure mereris. (19v) Anschließend nachgetragen V. 515–526: Si vetus exemplum non sufficit ecce novellum … Cordis ad exemplar regis formata Richardi. Vgl. Hinweis am Blattrand: Explicit hic liber sit scriptor crimine liber (Colophons Nr. 21008. Vgl. W. Wattenbach, Das Schriftwesen im Mittelalter. Leipzig 31896, 509 und 522). Hoc exem plum apostrophe debet sequi vel postponi illi exemplo. Ancillatur item etc. (19v) Schreibervers: Finito libro grates Christo referamus. Darunter Schlussschrift: Explicit poetria novella Ganfredi. Lit.: Guizzardo da Bologna, Recollecte super Poetria magistri Gualfredi, a cura di D. Losappio (Gli umanisti 3). Verona 2013; Woods 144–145, 299, mit Hinweis auf die Verwandtschaft des Kommentars von Cod. 327 mit Rom, Vaticana, Cod. Ottob. Lat. 3291. 2 (20ra–21vb) ‹Vitalis Blesensis›: Geta, unvollständig (Ed. Paeske, mit Nennung der Hs. F. B ertini, La comedia elegiaca latina in Francia nel secolo XII. Genova 1973, 90–120. – Walther, Initia Nr. 7272). Tit.: Incipit Geta. – Inc.: Grecorum studia nimiumque diuque secutus … – Expl. mut. (V. 444): … Si quecumque loquor omnia falsa forent [ (Ir–v) leer. P. K. Cod. 353 LIBER HORARUM, Bd. 1 (zusammengehörend mit Cod. 654) Perg. VI, 194 Bl. 160 × 120. Raum Brixen, um 1496. B: Regelmäßiges Perg., vereinzelt kleine Löcher und Nahtspuren. Lagen: (IV–2)VI + 2.V20 + IV28 + 10.V128 + IV136 + 5.V187(186) + (V–2)195(194). Gegenbl. zu Bl. I (wohl ohne Textverlust) und Bl. II (Textverlust) sowie zu Bl. 188 und 189 (Textverlust), zudem wohl auch einstiges Nachsatzbl. herausgerissen bzw. -geschnitten. Am Ende der Lagen 3 und 5–8 stark beschnittene Reklamanten. Um die zweite Lage schmaler Pergamentfalz geschlagen. Am oberen Blattrand zeitgenössische rote Foliierung in arabischen Ziffern 1–195 (hier berück sichtigt); Fehler in der Zählung: 153 doppelt angeführt, 164 und 168 ausgelassen. Vorgebundenes Kalendar von späterer Hand in römischen Zahlen foliiert I–VI. Inhaltsverzeichnis (Bl. Iv) mit Folioangaben bezieht sich auf Cod. 353 (= Bd. 1) und 654 (= Bd. 2, hier ohne Folioangaben). S: Bl. 1r–195v Schriftraum 110 × 70, von jeweils zweifachen roten Tintenlinien gerahmt, zu 18–20, größten teils 19 Zeilen auf feiner Bleistift-, tw. Blindlinierung. Zirkelstiche. Kalligraphische Bastarda (auffällig gelängte Initialmajuskeln, Unterlängen und Kürzungsstriche, bisweilen i-Punkte in Kreisform). Schreiber: Georg Hölzl aus Mattighofen, um 1496, vgl. Cod. 654 (Bl. 202r: dat. 1496). Kalendar Bl. IIr–VIv Schriftraum 115 × 75/85. Zwei Spalten, von roten Tintenlinien gerahmt. Inhaltsver zeichnis Bl. Ir–v Schriftraum 115 × 75, von jeweils zweifachen roten Tintenlinien gerahmt, Folioangaben im Inhaltsverzeichnis von Hand der Foliierung (wie Cod. 654). Kalligraphische Bastarda. Schreiber: Georg Hölzl aus Mattighofen. Nachträge von späterer Hand. A: Rote Auszeichnungsstriche, (Zwischen-)Überschriften, Interpunktionszeichen, Unterstreichungen; wenige, relativ große rote und blaue Paragraphzeichen. Bl. Ir am Textbeginn größere rote Cadelle; Bl. 102r schwarze Cadelle mit Federzeichnungsdekor. Ein- bis sechszeilige Lombarden (als KL-Ligaturen zweizeilig), ab wechselnd hell-/dunkelrot und blau, mit Punktverdickungen bzw. gestielten Punkten; Bl. 10v aufwändige dreizeilige blaue Lombarde mit markanten Punktverdickungen, freiem Punkt und in Achterschlingen geleg ten Ausläufern, gut vergleichbar mit den Lombarden in Brixen, Priesterseminarbibl., Cod. C.8 (ab Bl. 157r), F.5, F.6 (z. B. Bl. 2v: wohl von demselben Lombardenmaler) und F.7; Bl. 94v Lombarde nicht ausgeführt. Repräsentanten. Deckfarbenschmuck: neun drei- bis sechszeilige Ornamentalinitialen mit Ranken, acht ganzseitige (Schrift spiegelgröße) rankenlose Miniaturen. 252 Cod. 353 Zu den Charakteristika der ornamentalen Initialen (Bl. 1r, 40r, 55r, 71r, 94r, 116r, 131r, 165r, 184r) s. Cod. 654; in Cod. 353 im Verhältnis mehr blattvergoldete Binnenfelder und Buchstabenkörper als in Cod. 654, zu den Punzierungen s. u.; bei goldenen Buchstabenkörpern die Blattkonturen in Rot eingetragen; Bl. 40r Buchstabenkörper (Deckfarbe) mit Kreisen gefüllt. – Randschmuck: Akanthusranken (Charakteristika s. Cod. 654) und einige naturalistische Zweige, die recht grob gemalt sind: Bl. 94r roter Klee, 165r Eichen zweige (mit Eicheln), 184r Weinranken; nur eine Drolerie (Bl. 1r): Dompfaff, der auf einem Bein steht. Zu den Charakteristika der Miniaturen (u. a. Figurenbildung, Hintergründe, Kolorit) s. Cod. 654. Bl. 39v Schmerzensmann: Christus, dessen Körper von Blutstropfen bedeckt ist, präsentiert stehend seine Wundmale; zwei flankierende Engel halten seinen roten Mantel (innen grün) auf; kobaltblauer Hintergrund (s. u. bei Punzierungen). Bl. 54v Letztes Abendmahl an einem runden Tisch; Johannes kauert auf dem Schoß Jesu; Judas, aus dessen Mund ein Teufelsfigürchen entweicht, mit rotbraunem Haar, ockergelbem Gewand und rotem Geldbeutel an der Seite; Hintergrund: Fenster (mit gemauertem Fensterkreuz), Landschaftsausblick. Vgl. kleinformatigere und daher einfachere Komposition in Cod. 654, Bl. 163v. Bl. 70v Dreifigurige Kreuzigung: am Fuß des grob gemaserten Kreuzes Steinhaufen und Gebeine Adams; Gekreuzigter mit gelbgrünem Lendentuch (vgl. Cod. 654, Bl. 15v und 165r oben); Kreuzestitel INRI (mit stark eingerollten Enden) auf das Kreuz aufgesteckt. Maria, die ihren Mantel vor dem Leib rafft, richtet den Blick verloren zu Boden, während Johannes mit gefalteten Händen in die Ferne blickt; Hintergrund: Stadt vedute (u. a. türmchenbesetzte Kirchenfassade), Fluss oder See und Bergkette. Bl. 93v Pietà flankiert von Maria Magdalena (neben ihr das Salbgefäß) und Johannes; auf Marias Herz weist großes Schwert als Zeichen ihres Schmerzes; Hintergrund: Kreuz (mit aufgestecktem Kreuzestitel), an dessen Querbalkenenden Geißel und Rute hängen, mittig Dornenkrone; Leidenswerkzeuge jeweils mit Blutstropfen benetzt. Bl. 115v Gnadenstuhl: Gottvater sitzt auf einer (Holz-?)Bank, hinter der zwei Engel ein blaues, mit golde nen Ähren übersätes Tuch hochhalten, und präsentiert mit zur Seite gerichtetem Blick das Kruzifix; darüber (auf einem in Rot eingezeichneten, stilisierten Wolkenband) Taube des hl. Geistes auf Goldgrund (Punzie rung s. u.); Wasserschaden (?), s. insbes. über der rechten Schulter Gottvaters. Bl. 130v Mondsichel-Madonna: Maria, über die zwei Engelchen eine Bügelkrone halten, steht auf hell blauem Sichelmond mit Gesicht (nach unten gerichtet). Das Jesuskind, das den linken Arm um den Hals seiner Mutter gelegt hat, präsentiert mit der Rechten einen roten Apfel (?); Mutter und Kind blicken einander an. Blattvergoldeter Hintergrund (Punzierung s. u.), Wiesengrund mit Maiglöckchen und rotblütiger Pflanze im Vordergrund. Bl. 163v Mariä Himmelfahrt: Apostel (zuvorderst Rückenfigur, deren Heiligenschein tellerartig am Hinter kopf sitzt) in einem Talkessel, u. a. mit hochgewachsenen Maiglöckchen; über ihnen wird Maria (mit gefalte ten Händen) von fünf Engeln, die einen Kranz um sie bilden, in den Himmel getragen; einer der Engel dient ihr als Fußstütze, direkt über ihrem Kopf rot-goldene Wolke, die Strahlen aussendet. Bl. 183v Verkündigung: Maria kniet betend an einem Pult, während der Engel (mit Pfauenflügeln und Spruchband AVE GRACIA PLEN[A]) segnend vor ihr niederkniet; hoch über ihnen zwei Fenster (über schnitten von einem blau ausgemalten Rippengewölbe, vgl. Cod. 654, Bl. 51v), das rechte vergittert, im linken Büste des segnenden Gottvaters, von dem goldene Strahlen ausgehen, auf welchen das nimbierte, Kreuz tragende Jesuskind und die Taube des hl. Geistes zu Maria hinabgleiten. Punzierungen: 1) Binnenfelder: Bl. 1r und 184r Rautengitter aus winzigen Quadraten, Bl. 1r mit fünfblätt rigen Blüten (aus kleinen Kreisen) gefüllt, Gitterschnittpunkte zudem mit vierblättrigen roten und blauen Blüten besetzt; Bl. 184r Rautengitter mit sechsblättrigen Blüten (aus kleinen Quadraten) gefüllt; hier auch die blattvergoldete Rankenschlinge punziert (Rautengitter mit Kreisen). – 2) Hintergründe von Miniaturen: Bl. 39v kobaltblaue Deckfarbe mit wie in einen Blattgoldgrund eingepunztem Rautengitter, darin sechsblätt rige Blüten; Bl. 115v Goldgrund mit größeren sechs- und kleineren fünfzackigen Sternen bzw. Sternblüten (auch auf Nimben); Bl. 130v Goldgrund mit größeren sechsblättrigen Blüten, die abwechselnd rot und blau ausgemalt sind, sowie kleineren fünfzackigen Sternen bzw. Sternblüten. Der eher grobe Deckfarbenschmuck stammt von der Hand, die auch Cod. 654 ausgestattet hat, in Cod. 353 allerdings etwas sorgfältiger und detaillierter (z. B. bei der Gestaltung der Fußböden, Gewänder, Möbel- und Architekturzier) zu Werk gegangen ist. Zur Einordnung s. bei Cod. 654. S. R. E: Renaissance-Einband: braunes Leder auf Holz mit Golddruck. Neustift, zw. 1569 und 1581 (identisch mit Cod. 654). Werkstatt: Narziß Schuechl. Einband 2014 restauriert (vgl. Restaurierbericht, drei kleine abgelöste Makulaturfragmente beigelegt). VD und HD gleich: ein von je zweifachen Streicheisenlinien in Gold begrenzter Rahmen: Rolle Enthauptung des Johannes, Eherne Schlange, Kreuzigung, Auferstehung (Nr. 1 = Neuhauser, Rolleneinbände, R 29). Im Mit Cod. 353 253 telfeld des VD Plattenstempel mit Darstellung eines Erzengels (Michael?) mit Wappenschild von Propst Augustin Schabl (Propst 1569–1581) mit zwei steigenden geflügelten Löwen (Nr. 2 = Neuhauser P 3). Da rüber Inschrift Augustinus praepositus Novæcelle. Im Mittelfeld des HD in den Ecken Einzelstempel mit floralem Dekor aus Vase wachsend (Nr. 3), in der Mitte Einzelstempel Kreuzigung Christi (Nr. 4). Zwei Leder-Metall-Schließen, beim Materialwechsel sich stark verjüngend. Kanten gerade und nach innen hin abgeschrägt und durch einfache Streicheisenlinie eingefasst. Rücken: vier Doppelbünde, von zweifachen goldenen Linien gesäumt, Linien am VD und HD fortgesetzt, spitz aufeinander zulaufend. In den Feldern zw. den Bünden jeweils florale Rolle (Nr. 5 = Neuhauser, Rolleneinbände, R 26). Im zweiten Feld Signatur schild (s. bei G). Kapitale mit gelbem und rotem Zwirn umstochen. Schnitt goldfarben und punziert. Spiegel des VD Perg. mit Exlibris, Signaturen und Stempel (s. bei G), Spiegel des HD leeres Papierblatt. G: Hs. im Auftrag des Kaspar Neuhauser, bischöflicher Zöllner (1490/92) und Stadtrichter (1494–1501) in Klausen (vgl. Kustatscher I 300–302 und II 646; K. Brandstätter, Städtische Verwaltung und Bürgerschaft in Klausen im 15. Jh. Der Schlern 73 [1999] 442, 718), 1496 von Georg Hölzl geschrieben. Hs. spätestens seit der Zeit des Propstes Augustin Schabl (1569–1581) in Neustift. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II1 E 14 (Tinte, Bleistift) sowie 353 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 353. Bl. Ir Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Am Spiegel des VD Stempel der Soprintendenza Tridentina 20. Jh. Restaurierbericht 2014 von A. Al Shami. L: Wilhelm I 135 und II 110. – Chev. Nr. 30585 und 30053 (ohne Nennung der Sign.). – AH 30, Nr. 8; 30, Nr. 12; 30, Nr. 52 und 31, Nr. 5. – Laußermayer 301–302, 309–310. – Neuhauser, Neustift 85. – Peintner, Buchmalerei, 16, 34, 84–86, 88–90 (ohne Nennung der Sign.). – Katalog ULB Tirol IV 217, 471. – Kustat scher I 302 (ohne Nennung der Sign.). – Kustatscher–Korenjak 157 (ohne Nennung der Sign.). Liber Horarum, Bd. 1. (Ir–v) Titel samt Inhaltsverzeichnis. Tit.: Subscripti cursus sparsim hincinde in devocionum libellis inventi summa cum diligencia hic in unum sunt collecti ut quivis excitandam devocionem prout suus afficitur animus ad manum habeat quo se orando convertat: aut scilicet ad trinitatem beatissimam aut ad eternam sapienciam ad corpus Christi ad ipsius passionem vel eius animam ad spiritum sanctum vel beatissimam virginem Mariam ad beatos angelos vel omnes Dei sanctos aut pro misericordia inpetrandam pro peccatis propriis vel fidel ibus defunctis cum septem penitencialibus psalmis cum mortuorum vigiliis et accessu altaris prout secundum ordinem huiusmodi cum numero foliorum hic sunt intitulati. Darunter in Schwarz Auflistung aller Kapitel samt Angabe der Blattzahl. Aufgelistet werden auch jene Kapitel, die in Cod. 654 zu finden sind, allerdings ohne Angabe einer Blattzahl. Inc.: Cursus de proprio angelo folio 1 … – Expl.: … Vespere mortuorum folio. 1 (IIra–VIv) Kalendar, unvollständig (bis 22. Dezember). Besonderheiten, die auf eine Entstehung in der Diözese Brixen hinweisen: Ingenuinus et Al buinus (5. Februar), Translatio s. Albuini (13. Mai), Dedicatio ecclesiae Brixinensis (31. Juli), Cassianus (13. August). (IIIra) Nachtrag am 15. März von späterer Hand zum Tod des Nikolaus Neuhauser, Kirchpropst (1483/84), Bürgermeister (1488) und Stadtrichter (1492) in Klausen (s. bei G): Anno domini 1496 obit Nicolaus Neünhausser pater meus dilectus. (IVrb) Nachtrag am 20. Juni von späterer Hand zum Tod des Bischofs Georg Golser: Anno domini 1489 obit Icorius (!) Golser episcopus Prixinensis dominus meus generosus. Fehlstelle: Nach Bl. VI zwei Blätter herausgerissen (s. bei B), auf erstem Blatt urspr. Forts. und Abschluss des Kalendars, tw. erhalten. 2 (1r–12v) Cursus de proprio angelo. Hymnen (AH 50, Nr. 146, Strophen auf die einzelnen Tages zeiten aufgeteilt, Str. 4 mehrfach), Antiphonen (De s. Michaele). Tit.: Incipit cursus de proprio angelo qui dicendus est dominica die. 254 Cod. 353 (1r) Invitatorium mit Ps 95. – (2r) Matutin. Hymnus. – (2v) Ps 8,5–9. – Antiphon (CAO Nr. 1491). – (3r) Leccio prima. Inc.: Stetit angelus iuxta aram templi habens thuribulum aureum in manu sua … (vgl. Apc 8,3). – Expl.: … salus honor virtus et potestas omnipotenti Deo. Tu autem. – Responsorium mit Versikel. – Te Deum. (4r) Laudes. Ps 148,1–5. – (4v) Antiphon (CAO Nr. 2102). – Capitula. – Hymnus. – (5r) Psalmus (recte: Lc 1,68–79). – (5v) Antiphon (CAO Nr. 1472). – Oracio. Deus qui summa providencia humano generi supernorum spirituum presidia subministras … (5v) Prim. Hymnus. – (6r) Ps 97,7–10. – Antiphona. Emitte angelum de celis ut me perducat in paradisum exultacionis alleluia. – Capitulum. – (6v) Responsorium mit Versikel. – Oracio. Omnipotens et misericors Deus qui hominem ad imaginem tuam formasti et ei angelum corporis et anime custodem … (7r) Terz. Hymnus. – Ps 103, 20–22. – (7v) Antiphona. Emitte angelum de celis ut me perducat in paradisum exultacionis alleluia. – Capitulum. – Responsorium mit Versikel. – Oracio. Omni potens et misericors Deus qui electos spiritus celestium gaudiorum … (8r) Sext. Hymnus. – Ps 104,1–4. – (8v) Antiphona. Emitte angelum de celis ut me perducat in paradisum exultacionis alleluia. – Capitulum. – Responsorium mit Versikel. – (9r) Oracio. Deus qui nos miseros peccatores perire non sinis presta quesumus ut angelum michi deputatum … (9r) Non. Hymnus. – (9v) Ps 143,10–12. – Antiphona. Emitte angelum de celis ut me perducat in paradisum exultacionis alleluia. – Capitulum. – Responsorium mit Versikel. – (10r) Oracio. Perpetuum nobis domine tue miseracionis presta subsidium … (10r) Vesper. Ps 138,1–3. – Antiphon (CAO Nr. 3757). – (10v) Capitulum. – Hymnus. (11r) Can ticum Mariae (nur Verweis) mit Antiphon (CAO Nr. 3755). – Oracio. Omnipotens sempiterne Deus qui angelum tuum antequam nasceretur homo et ego nascerer ad custodiendum … (11v) Komplet. Ps 91,10–12. – Antiphona. Emitte angelum de celis ut me perducat in paradisum exultacionis alleluia. – Hymnus. – (12r) Capitulum mit Versikel. – Canticum Simeonis (nur Verweis) mit Antiphon (CAO Nr. 3592). – (12v) Oracio. Deus qui fidelium tuorum deprecacio nem semper exaudis presta michi indigno peccatori … 3 (13r–25v) Cursus de aeterna sapientia (Druck: u. a. H. Porro, Vita e morte della sig. Cornelia Lampugnana … Pavia 1624, 215–240. Henricus Suso zugewiesen, vgl. P. Künzle, Heinrich Seuses Horologium Sapientiae. Erste kritische Ausgabe unter Benützung der Vorarbeiten von D. Planzer OP [Spicilegium Friburgense 23]. Freiburg 1977, 606ff.). Hymnen (Mone I 329 Nr. 258). Tit.: Cursus de eterna sapiencia feria secunda dicendus. (13r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus. – (13v) Ps 66,1–4. – Antiphon (CAO Nr. 4810) mit Versikel. – (14r) Benediccio. Precibus et meritis sanctissime sue matris salvat et protegat … – Leccio prima. Inc.: Sapiencia eterna splendor glorie et substancia patris qui universa de nichilo creasti … – Expl.: … sed tu in me et ego in te indissolubili vinculo amoris eternaliter maneamus. Tu autem. – (14v) Responsorium mit Versikel. – Leccio secunda. Inc.: Ihesu mi dulcissime felicissima sapiencia verbum patris principium et finis alpha et o omnium encium … – Expl.: … sed permaneat morte forcior noster amor in evum. – (15r) Responsorium mit Versikel. – (15v) Leccio tercia. Inc.: O refugium meum o liberator meus per immensum amorem qui te in patibulo crucis amarissimam mortem pro me subire coegit … – Expl.: … et ad conspectum glorie tue pervenire concedas. Tu autem. – (16r) Responsorium mit Versikel. (16r) Laudes. Ps 117. – (16v) Antiphon (CAO Nr. 4811). – Capitulum. – Hymnus. – (17r) Ps. (recte: Lc 1,68–79). – (17v) Antiphon (CAO Nr. 4081). – (18r) Oracio. Deus qui per coeternam sapienciam hominem cum non esset condidisti … (18r) Prim. Hymnus. – (18v) Ps 143,8–10. – Antiphona. Ego diligentes me diligo et qui mane vigilaverint ad me invenient me. – Capitulum. – (19r) Responsorium mit Versikel. – (19r) Oracio. Corda nostra quesumus domine eterne sapiencie splendor illustret … Cod. 353 255 (19r) Terz. Hymnus. – Ps 143,10–12. – (20r) Antiphona. Filii concupisce sapienciam conserva iusticiam et prebebit eam dominus tibi. – Capitulum. – Responsorium mit Versikel. – Oracio. Deus qui per coeternam sapientiam hominem cum non esset condidisti … (20v) Sext. Hymnus. – (21r) Ps 33,20–22. – Antiphon (CAO Nr. 2418). – Capitulum. – Respon sorium mit Versikel. – (21v) Oracio. Exaudi nos misericors Deus et mentibus nostris sapiencie tue lumen ostende … (21v) Non. Hymnus. – (22r) Ps 51,12–14. – Antiphon (CAO Nr. 2418). – Capitulum. – (22v) Responsorium mit Versikel. – Oracio. Infunde quesumus domine Deus tue cordibus lumen sa piencie … (22v) Vesper. Ps 111,9–10. – (23r) Antiphon (CAO Nr. 4153). – Capitulum. – Responsorium mit Versikel. – Hymnus. – (23v) Versikel. – Magnificat (nur Verweis). – Antiphon (CAO Nr. 4050). – Oracio. Deus qui per coeternam sapienciam hominem cum non esset condidisti … (24r) Komplet. Ps 13,4–6. – Antiphon (CAO Nr. 2576). – (24v) Capitulum. – Responsorium mit Versikel. – Hymnus. – (25r) Canticum Simeonis (nur Verweis). – Antiphon (Cantus Nr. 203570). – (25v) Oracio. Fragilitatem nostram quesumus domine benignus respice et saporem nobis eterne sapiencie … 4 (26r–39r) Cursus de sancto spiritu. Hymnen (falls nicht anders ausgewiesen AH 50, Nr. 144). Tit.: Cursus de sancto spiritu tercia feria dicendus. (26r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus (AH 54, Nr. 153). – (26v) Ps 1. – (27r) Anhtiphon (CAO Nr. 5327). – Benediccio. Confortare et corroborare in bonis operibus … – (27v) Leccio (2 Cor 13,13). – Responsorium mit Versikel. (27v) Laudes. Ps 93. – (28r) Antiphon (CAO Nr. 5005). – Capitulum. – Hymnus (Chev. Nr. 2340). – (29r) Ps. (recte: Lc 1,68–79). – (29v) Antiphon (CAO Nr. 4920). – Oracio. Concede quesumus omnipotens Deus sanctum nos spiritum vocis promereri … – (30r) Antiphon De sancta trinitate (CAO Nr. 5119). – Oracio. Omnipotens sempiterne Deus qui dedisti famulis tuis in con fessione vere fidei eterne trinitatis gloriam agnoscere … – (30v) Antiphon De beata virgine (CAO Nr. 1563). – Oracio. Deus qui virginalem aulam beate Marie virginis in qua habitares eligere dignatus es … – (31r) Antiphon De omnibus sanctis (CAO Nr. 4726). – Oracio. Protege domine populum tuum et apostolorum tuorum Petri et Pauli et omnium aliorum … – Oracio. Omnes sancti tui quesumus domine nos ubique letificent et adiuvent ut dum eorum merita reco limus patrocinia senciamus … (31v) Prim. Hymnus. – (32r) Ps 54,3–9. – (32v) Antiphon (CAO Nr. 5327). – Capitulum. – Res ponsorium mit Versikel. – (33r) Oracio. Cordibus nostris quesumus domine spiritum sanctum … (33r) Terz. Hymnus. – (33v) Ps 70. – (34r) Antiphon (CAO Nr. 5327). – Capitulum. – Responso rium mit Versikel. – Oracio etc. Mentes nostras quesumus domine spiritus sanctus reparet … (34r) Sext. – (34v) Hymnus. – Ps 87. – (35r) Antiphon (CAO Nr. 5327). – Capitulum. – Respon sorium mit Versikel. – Oracio. Deus cui omne cor patet et omnis voluntas loquitur et quem n ullum latet secretum … (35v) Non. Hymnus. – Ps 124. – (36r) Antiphon (CAO Nr. 5327) (nur Verweis). – Capitulum. – Responsorium mit Versikel. – Oracio. Illo nos igne spiritus sanctus inflammet quem dominus noster Ihesus Cristus … (36v) Vesper. Ps 128,1–5. – Antiphon (CAO Nr. 5327). – Capitulum. – (37r) Responsorium mit Versikel. – Hymnus. – (37v) Magnificat (nur Verweis). – Antiphon (CAO Nr. 4902). – (38r) Ora cio. Deus qui corda fidelium sancti spiritus illustracione docuisti … (38r) Komplet. Ps 129. – (38v) Antiphon (CAO Nr. 5327) (nur Verweis). – Hymnus (nur Ver weis). – Capitulum. – (39r) Canticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 5005). – Oracio. Sancti spiritus corda nostra mundet infusio … (39v) ganzseitige Miniatur (s. bei A). 256 Cod. 353 5 (40r–54r) Cursus de misericordia dni. Hymnen (Chev. Nr. 30585. AH 30, Nr. 12). Tit.: Cursus de misericordia domini quarta feria dicendus. (40r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus. – (40v) Ps 25,6–9. – (41r) Bene diccio. Ostende nobis domine misericordiam tuam et salutare tuum da nobis … – Leccio prima (Sap 11,23–26). – Responsorium mit Versikel. – (41v) Leccio secunda (Sir 36,1–5). – (42r) Responsorium mit Versikel. – Leccio tercia (Dn 3,31,29,30,41–43). – (42v) Responsorium mit Versikel. – Te Deum. Inc.: Te Deum laudamus te dominum misericordissimum confitemur tuam maximam misericordiam semper imploramus … – Expl.: miserere miserere miserere nostri Ihesu benigne: qui passus es pro nobis clementer in cruce Amen. (44r) Laudes. Ps 25,15–18. – (44v) Antiphon (vgl. CAO Nr. 3816). – Capitulum. – Hymnus. – (45r) Canticum Zachariae. – (45v) Antiphona. Cum vidisset dominus viduam misericordia motus super illam dixit et noli flere. – Oracio. Deus qui omnipotenciam tuam parcendo maxime et miserando manifestas multiplica super nos misericordiam tuam … (46r) Prim. Hymnus. – Ps 51,3–5. – (46v) Antiphona. Euntes discite quid est misericordiam volo et non sacrificium: non enim veni vocare iustos sed peccatores dicit dominus. – Capitulum. – Res ponsorium mit Versikel. – (47r) Oracio. Subveniat nobis domine misericordia tua ut ab immi nentibus peccatorum nostrorum periculis … (47r) Terz. Hymnus. – (47v) Ps 119,75–78. – (48r) Antiphon CAO Nr. 2368). – Capitulum. – Res ponsorium mit Versikel. – Oracio. Pateant aures misericordie tue domine precibus s upplicancium et ut petentibus desiderata … (48v) Sext. Hymnus. – Ps 69,14–17. – (49r) Antiphon (CAO Nr. 3873). – Capitulum. – (49v) Res ponsorium mit Versikel. – Oracio. Adesto supplicacionibus nostris omnipotens Deus et q uibus fiduciam sperande pietatis indulges … (49v) Non. Hymnus. – (50r) Ps 103, 8–11. – Antiphon (CAO Nr. 3770). – (50v) Capitulum. – Responsorium mit Versikel. – Oracio. Tua nos domine Deus misericordia ab omni surrepcione vetustatis … (51r) Vesper. Ps 123, 1–3. – Antiphon (CAO Nr. 4695). – Capitulum. – (51v) Responsorium mit Versikel. – Hymnus. (52r) Magnificat (nur Verweis). – Antiphona. Cum adhuc longe esset filius vidit illum pater suus et misericordia motus est occurrens cecidit super collum eius et osculatus est eum. – Oracio. Omnipotens sempiterne Deus qui ex abundancia pietatis tue et merita suppli cum excedis … (52r) Komplet. – (52v) Ps 86,3–7. – Antiphona. Procedens servus dominum rogabat dicens pacienciam habem me et omnia reddam tibi miserttus autem dominus servi illius dimisit eum et debitum dimisit ei. – Hymnus. – (53r) Capitulum. – Responsorium mit Versikel. – (53v) Canti cum Simeonis. – Preces. Ego dixi miserere mei domine. Sana animam meam quia peccavi tibi. Convertere domine … – (54r) Oracio. Protector noster in te sperancium Deus sine quo nichil est validum … (54v) ganzseitige Miniatur (s. bei A). 6 (55r–70r) Cursus de corpore Christi. Tit.: Cursus de corpore Christi quinta feria dicendus. (55r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus (AH 50, Nr. 388). – (55v) Ps 16. – (56v) Antiphon (Cantus Nr. 200833). – Benediccio. Corporis et sanguinis domini nostri Ihesu Cristi sacramentum sit … – Leccio prima. Inc.: Immensa divine largitatis beneficia exhibita po pulo Christiano inestimabilem ei conferunt dignitatem … – Expl.: … servitute a peccatis omni bus mundaremur. Tu autem. – Responsorium mit Versikel. – (57v) Leccio secunda. Inc.: Et ut tanti beneficii iugis in nobis maneret memoria corpus suum in cibum … – Expl.: … de acciden tibus iudicant sibi notis. Tu autem. – (58r) Responsorium mit Versikel. – (58v) Leccio tercia. Inc.: Nullum eciam sacramentum est isto salubrius quo purgantur peccata … – Expl.: … Deo gra ciarum debite acciones. Tu autem. – (59v) Responsorium mit Versikel. – Te Deum (nur Verweis). Cod. 353 257 (59v) Laudes. Ps 20,2–7. – (60r) Antiphon (Cantus Nr. 203078). – Capitulum. – Hymnus (AH 50, Nr. 395). – (60v) Canticum Zachariae (nur Verweis). – Antiphon (CAO Nr. 2594). – Oracio. Deus qui nobis sub sacramento mirabili passionis tue memoriam reliquisti tribue … (61r) Prim. Hymnus (Chev. Nr. 23597). – (61v) Ps 23. – (62r) Antiphon (Cantus Nr. 203765). – Capitulum. – Responsorium mit Versikel. – Oracio. Reminiscere miseracionum tuarum quesu mus domine et famulos tuos … (62v) Terz. Hymnus (Chev. Nr. 23749). – (63r) Ps 78,23–30. – Antiphon (Cantus Nr. 200280). – (63v) Capitulum. – Responsorium mit Versikel. – Oracio. Deus qui ecclesiam tuam preciosi corporis et sanguinis domini nostri Ihesu Christi misterio … (64r) Sext. Hymnus (Chev. Nr. 7807). – Ps 81, 9–17. – (64v) Antiphon (Cantus Nr. 200786). – (65r) Capitulum. – Responsorium mit Versikel. – Oracio. Propitiare nobis quesumus omnipotens pater per unigeniti tui corpus et sanguinem … (65r) Non. Hymnus (Chev. Nr. 33769). – (65v) Ps 84,2–5. – (66r) Antiphon (Cantus Nr. 201674). – Capitulum. – Responsorium – Oracio. Domine Ihesu Christe fili Dei vivi qui pro nobis in cruce moriens dulcissimi lateris … (66v) Vesper. Ps 128. (67r) Antiphon (Cantus Nr. 204661). – Capitulum. – Responsorium mit Versikel. – Hymnus (vgl. Chev. Nr. 27686). – (68r) Magnificat (nur Verweis). – Antiphon (Cantus Nr. 203576). – Oracio. Domine Ihesu Christe qui in ultima cena tradidisti discipulis tuis … (68r) Komplet. – (68v) Ps 4,6–9. – (69r) Antiphon (Cantus Nr. 200020). – Hymnus (Chev. Nr. 28563). – (69v) Capitulum. – Canticum Simeonis. – Antiphon (Cantus Nr. 203492). – (70r) Oracio. Deus cuius unigenitus tuus pro nobis usque ad mortem obediens … (70v) ganzseitige Miniatur (s. bei A). 7 (71r–93r) Cursus de passione dni. Hymnen (AH 50, Nr. 382). Tit.: Cursus de passione domini sexta feria dicendus. (71r) Invitatorium mit Ps 95. – Matutin.– (71v) Hymnus. – (72r) Ps 2. – (73r) Antiphona. Insur rexerunt in me viri absque misericordia et non pepercerunt anime mee. – Benedictio. Gloriosa passio Christi perducat nos ad gaudia paradisi … – (73v) Leccio prima (Io 19,1–3). – Respon sorium mit Versikel. – (74r) Leccio secunda (Io 19, 16–17). – Responsorium mit Versikel. – Leccio tercia (Io 19, 28–30). – Responsorium mit Versikel. – (74v) Te Deum. (75r) Laudes. Ymnus (recte: Ps 13). – (75v) Antiphon (CAO Nr. 1913). – Capitulum. – (76r) Hymnus. – (76v) Canticum Zachariae. – (77r) Antiphon (CAO Nr. 4395). – Oracio. Domine Ihesu Christe fili Dei vivi qui hora matutina stans ligatus … – (77v) Alia oracio. O virgo virgi num sanctissima Dei genitrix Maria propter gravamen et tormentum quo torquebatur … (78r) Prim. Hymnus. – Ps 43. – (78v) Antiphon (CAO Nr. 2833). – Capitulum. – Responsorium mit Versikel. – (79r) Oracio. Domine Ihesu Criste fili Dei vivi qui hora diei prima permisisti te duci in consilio Iudeorum … – (79v) Alia oracio. O virgo virginum predulcissima mater Maria quando vidisti filium tuum … (80r) Terz. Hymnus. – (80v) Ps 64. – (81r) Antiphon (CAO Nr. 2422). – Capitulum. – Responso rium mit Versikel. – (81v) Oracio. Domine Ihesu Criste fili Dei vivi qui hora diei tercia fuisti ab Herode illusus … – Alia oracio. O virgo virginum beatissima virgo Maria propter angustias et cruciam inaque cor tuum sustinuit … (82v) Sext. Hymnus. – Ps 116,10–19. (83r) Antiphon (CAO Nr. 4343). – Capitulum. – (83v) Responsorium mit Versikel. – Oracio. Domine Ihesu Criste qui hora diei sexta ante crucem fuisti denudatus … – (84r) Alia oracio. O virgo virginum clementissima mater Maria propter doloris gladium qui pertransivit animam tuam … (84v) Non. Hymnus. – Ps 142. (85v) Antiphon (CAO Nr. 1970). – Capitulum. – Responsorium mit Versikel. – Oracio. Domine Ihesu Christe fili Dei vivi qui hora diei nona pendens in cruce et clamans voce magna te derelictum dixisti … – (86r) Alia oracio. O virgo virginum sanctissima regina propter gravamen et tormentum quo torquebatur spiritus tuus iuxta crucem … 258 Cod. 353 (87r) Vesper. Ps 30. – (88r) Antiphona. Dignum es domine Deus noster accipere librum et solvere singnacula (!) eius quoniam occisus es et redimisti nos Deus noster. – Capitulum. – Respon sorium mit Versikel. – (88v) Hymnus. – (89r) Magnificat (nur Verweis). – Antiphona. Recessit pastor noster bonus fons aque vive qui posuit animam suam pro ovibus suis et pro grege suo mori dignatus est. – Oracio. Domine Ihesu Criste fili Dei vivi qui hora vesperarum cenasti cum discipulis tuis turbatus spiritu unum ex eis te traditurum … – (89v) Alia oracio. O virgo virginum castissima Dei genitrix Maria propter planctum acerbi eiulatus … (90r) Komplet. Ps 88. – (91v) Antiphon (CAO Nr. 4295). – Hymnus. – Capitulum. – (92r) Can ticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 4690). – Oracio. Domine Ihesu Christe fili Dei vivi qui hora completorii sepultus es et a mulieribus planctus es et lamentatus … – (92v) Alia oracio. O virgo virginum gloriosissima mater Maria propter gemitus et suspiria indicibiliaque lamenta quibus afficiebaris … (93v) ganzseitige Miniatur (s. bei A). 8 (94r–115r) Cursus de compassione BMV, Papst Johannes XXII. zugeschrieben (vgl. Bl. 114v). (Druck: u. a. Thesaurus cursuum devotissimorum … collectus ab G. B. Pontano. Prag 1600, 185– 219). Tit.: Cursus de compassione Marie virginis sabatho dicendus. (94r) Invitatorium. – Psalmus (AH 31, Nr. 158). – Matutin. – (94v) Hymnus (Chev. Nr. 8483). – (95r) Ps 88,2–10. – Antiphona. Nocte qua Ihesus capitur absque somno fuisti que soporatis cete ris flens pervigil mansisti. – Ps 88, 5–7. – (95v) Antiphona. Hec nox merorifera tunc fit et amara matri quando capitur proles eius cara. – Ps 88,8–10. – (96r) Antiphona. Erat mater virgo pia plorans foris atriter regem mundi furibundi dum tractarent viliter. – Benediccio. Salutifera lamentacio Christi conferat nobis gaudia paradise … – Leccio prima (Mone II Nr. 441). – (96v) Responsorium mit Versikel. – Leccio secunda. Inc.: De cruce quando dominus ad cumulum por tatur confestim ei genitrix portantes … – Expl.: … rigans lacrimis madefecit. Tu autem. – (97r) Responsorium mit Versikel. – Leccio tercia (Mone II Nr. 443). – (97v) Te Deum. (98r) Laudes. Ps 22,2–16. – (98v) Antiphona. Virgo regis celici genitrix iugiter preconiis honore tur dignis. – Ps 22,5–7. – (99r) Antiphona. Nunquam mater pertulit tam anxios dolores ut Maria filii considerans livores. – Ps 22,8–12. – Antiphona. Nulla novit genitrix angustias tam duras plagas cum aspiceret et captis puncturas. – (99v) Ps 22,12–14. – Antiphona. Rubens color verti tur matris in palorem dum conspicit filii crurorem. – Ps 22,15–16. – (100r) Antiphona. Desperata creditur que culpam non commisit eius natus moriens dum spiritum emisit. – Capitulum. – Hym nus (AH 4, Nr. 87). – (100v) Canticum Zachariae. – (101v) Antiphona. Tanquam latro capitur funibus constrictus Christus virgis ceditur alapis afflictus … – Oracio. Domine Ihesu Christe clementissime qui pro nobis peccatoribus mortem subiens temporalem nec materno dolori … – Alia oracio. Domine Ihesu Christe fili virginis et matris Marie qui hora matutinali pro salute humani generis ex Maria … (102v) Prim. Hymnus (Chev. Nr. 9578). – (103r) Ps 22,17–19. – Antiphona. Quantam putas ma tris mesticiam cum sentiret nati tristiciam non est dolor … – Capitulum. – (103v) Responsorium mit Versikel. – Oracio. Domine Ihesu Christe qui pro nobis peccatoribus in cruce agonizans … – (104r) Oracio. Domine Ihesu Christe fili virginis Marie salus mundi qui hora diei prima Pylato presidi fuisti presentatus … (104v) Terz. Hymnus (Chev. Nr. 12570). – Ps 55,3–5. – (105r) Antiphona. Agnus mitis tormenta paciens: heli clamat pro nobis moriens … – Capitulum. – Responsorium mit Versikel. – (105v) Oracio. Domine Ihesu Christe qui pro nobis peccatoribus in cruce agonizans … – Oracio. Domi ne Ihesu Criste fili virginis Marie salus mundi qui hora diei tercia coram Pylato preside purpura indutus fuisti … (106r) Sext. Hymnus (Chev. Nr. 17058). – (106v) Ps 59,2–4. – Antiphona. Clamor Christi matrem angustiat nullus planctus plangentem faciat … – Capitulum. – Responsorium mit Versikel. – Cod. 353 259 (107r) Oracio (Verweis auf vorangegangenes Gebet). – Alia oracio. Domine Ihesu Christe fili virginis Marie salus mundi qui hora diei sexta in cruce alta clavis durissimis affixus fuisti … (107v) Non. Ymnus. Rerum Deus tenax vigor Christus in cruce moritur sed pena fit acerbior cum reus non compatitur … – (108r) Ps 69,20–21. – Antiphona. Postquam hoc diversorum est clau sum virgo filium liquit qui fecit seculum flendo sequens discipulum. – (108v) Capitulum. – Res ponsorium mit Versikel. – Oracio (Verweis auf vorangegangenes Gebet). – Alia oracio. Domine Ihesu Christe fili virginis Marie salus mundi qui hora diei nona crucifixus manibus et pedibus verberibus et flagellis … (109r) Vesper. Ps 69,2–3. – Antiphona. Omnis etas defleat mortem redemptoris virgini condoleat matri salvatoris. – Ps 69,4–5. – (109v) Antiphona. Virgo cum aspiceret natum in cruce mori: suo nullus similis dolor est dolori. – Ps 69,6–7. – Antiphona. Matris quis angustias poterat auferre cum in cruce filium cerneret clavari. – Ps 35,11–13. – (110r) Antiphona. Queso mihi misero vir ginum regina teccum (!) flendo lacrimas humiles propina. – Ps 109,1–4. – Antiphona. Fac ut ita defleam mortem tui nati partemque possideam regni reparati. – Capitulum. – (110v) Respon sorium mit Versikel. – Hymnus (Mone II 142, Nr. 439). – (111r) Magnificat (nur Verweis). – An tiphona. Expirante filio mortem mater indicat stat in plebis medio penas mortis publicat … – Oracio. Domine Ihesu Christe virginis fili dulcissime qui cum proditore cenasti … – (111v) Alia oracio. Domine Ihesu Christe fili virginis Marie salus mundi qui hora diei vespertina pressuris innumeris nimis fatigatus … (112r) Komplet. Ps 41,8–11. – (112v) Antiphona. Recolamus virginis gloriose luctus cum in tem plum dominus vinctus fuit ductus. – Capitulum. – Responsorium mit Versikel. – (113r) Ymnus. Tu lucis ante terminum Christe sepulcro clauderis te plorat virgo virginum Deus humani gene ris … – (113v) Canticum Simeonis (nur Verweis). – Antiphona. Tuam pertransivit animam virgo carens crimine Symeonis gladuis omni cum gravamine qui devote dixerat … – Oracio. Salvator clementissime qui inter opprobria que tulisti pie matris anxietatibus compassus fuisti … – (114r) Alia oracio. Christe rex clementissime fili Marie qui inter plurima opprobria sanctissimum spir itum emittendo patri tuo … – Antiphona. De beata virgine Salve regina. – Oracio. Interveniat quesumus domine Ihesu Christe pro nobis apud tuam ineffabilem clemenciam nunc et in hora mortis nostre … (114v–115r) Schlussschrift: Has horas composuit papa Iohannes XXII et donavit omnibus vere penitentibus confessis et contritis eas devote dicentibus quadraginta annos indulgenciarum et quatuor carenas. Anschließend (in Schwarz): Et sequitur cursus de sancta trinitate in secundo folio etc. (115v) ganzseitige Miniatur (s. bei A). 9 (116r–130r) Cursus de sancta trinitate. Tit.: Cursus de sancta trinitate qui es trinus et unus miserere nobis. (116r) Invitatorium mit Verweis auf Ps. 95. – Matutin. Hymnus (Chev. Nr. 487. AH 51, Nr. 96). – (116v) Ps 8. – (117r) Antiphona. Benedictus es Deus patrum nostrorum laudabilis et gloriosus et superexaltatus in secula. – Benediccio. Te invocamus te adoramus te laudamus beata trinitas miserere nobis … – (117v) Leccio prima. Inc.: Adesto Deus unus omnipotens pater et filius et spi ritus sanctus te unum in substancia trinum … – Expl.: … tibi graciarum accio in secula. Tu autem domine. – Responsorium mit Versikel. – (118r) Leccio secunda. Inc.: Deus gracias tibi vera una trinitas: una et trina unitas te sanctam trinitatem … – Expl.: … omnium creaturarum tibi laus tibi gloria tibi graciarum accio in secula. Tu autem. – Responsorium mit Versikel. – (118v) Inc.: Credimus te Deum patrem a temetipso non ab alio nec factum nec creatum nec genitum te filium … – Expl. … Libera nos salva nos iustifica nos o beata trinitas. Tu autem domine. – Res ponsorium mit Versikel. (119r) Laudes. Ps 100. – (119v) Antiphona. O beata trinitas et benedicta et gloriosa unitas pater et filius et spiritus sanctus tibi laus tibi Gloria tibi graciarum accio. – Capitulum. – Hymnus 260 Cod. 353 (Chev. Nr. 13381. AH 31, Nr. 5) – (120r) Canticum Zachariae. – (121r) Antiphon (CAO Nr. 1707). – Oracio. Omnipotens sempiterne Deus qui dedisti famulis tuis in confessione vere fidei eterne trinitatis gloriam agnoscere … (121r) Prim. Hymnus (Chev. Nr. 13150. AH 51, Nr. 40 und 2, Nr. 17). – (121v) Ps 24. – (122r) Antiphon (CAO Nr. 2948). – Capitulum. – Responsorium mit Versikel. – (122v) Oracio. Sancta trinitas unus et omnipotens Deus nos quesumus a peccatorum nostrorum maculis … (123r) Terz. Hymnus (Chev. Nr. 13150. AH 51, Nr. 40 und 2, Nr. 17). – Ps 67. – (123v) Antiphon (CAO Nr. 3601). – Capitulum. – Responsorium mit Versikel. – (124r) Oracio. Omnipotens sem piterne Deus qui in unitate permanes et in trinitate semper consistis … (124r) Sext. Hymnus (nur Verweis: Chev. Nr. 13150. AH 51, Nr. 40 und 2, Nr. 17). – Ps 117. – (124v) Antiphon (CAO Nr. 2947). – Capitulum. – Responsorium mit Versikel. – (125r) Oracio. Domine sancte pater omnipotens sempiterne Deus famulos tuos maiestati tue subiectos per uni cum filium tuum … (125r) Non. Hymnus (nur Verweis: Chev. Nr. 13150. AH 51, Nr. 40 und 2, Nr. 17). – Ps 113. – (125v) Antiphon (CAO Nr. 3600). – Capitulum. – (126r) Responsorium mit Versikel. – Oracio. Concede quesumus omnipotens Deus ut sicut in nomine patris et filii divini generis intelligimus … (126r) Vesper. Ps 147,1–11. – (127r) Antiphon (CAO Nr. 5120). – Capitulum. – Responsorium mit Versikel. – (127v) Hymnus (Chev. Nr. 487. AH 51, Nr. 96). – (128r) Magnificat (nur Verweis). – Antiphon (CAO Nr. 5117). – (128r) Oracio. Filium tuum omnipotens pater ante secula gemitum invocantes exoramus … (128v) Komplet. Ps 150. – (129r) Antiphon (CAO Nr. 4086). – Capitulum. – Responsorium mit Versikel. – Hymnus (Chev. Nr. 10968. AH 52, Nr. 117, 5). – (129v) Canticum Simeonis (nur Verweis). – Antiphona. Sancta trinitas unus Deus omnipotens exaudi clemens supplices tuos et solve vincula peccatorum nostrorum … – (130r) Oracio. Exaudi nos domine sancte pater omni potens eterne Deus invocacionem omnipotentis filii tui … (130v) ganzseitige Miniatur (s. bei A). 10 (131r–163r) Cursus communis de BMV. Tit.: Cursus de communi de beata Maria virgine. (131r) Invitatorium mit Verweis auf Ps. 95. – Matutin. Hymnus (Chev. Nr. 16347. AH 50, Nr. 72). – (131v) Ps 8. – (132r) Antiphon (CAO Nr. 1709). – Ps 19. – (133v) Antiphon (CAO Nr. 4942). – Ps 24. – (134r) Antiphon (CAO Nr. 1438). – Benediccio. Precibus et meritis beatis sime Marie virginis et omnium sanctorum perducat nos dominus ad regna celorum … – (134v) Leccio prima (Sir 24,11–13). – Responsorium mit Versikel. – (135r) Leccio secunda (Sir 24,15– 16). – Responsorium mit Versikel. – Leccio tercia (Sir 24,17–20). – (135v) Responsorium mit Versikel. – Te Deum (nur Verweis). (136r) Laudes. Ps 93. – (136v) Antiphon (CAO Nr. 1503). – Ps 100. – (137r) Antiphon (CAO Nr. 3707). – Ps 63. – (137v) Ps 67. – (138r) Antiphon (CAO Nr. 3261). – Psalmus (recte: Dn 3,57–88). (139v) Antiphon (CAO Nr. 1705). – Ps 148. – (140v) Ps 149.– (141r) Ps 150. – Anti phon (CAO Nr. 4418). – (141v) Capitulum. – Hymnus (Chev. Nr. 16347. AH 50, Nr. 72, 6–8). – Psalmus (recte: Lc 1,68–79). – (142v) Antiphon (CAO Nr. 4029). – (143r) Oracio. Concede nos famulos tuos quesumus domine Deus noster perpetua mentis et corporis sanitate gaudere … – Compassio: Hora matutina Marie nuncciatur … (vgl. AH 30, Nr. 46 und Walther Nr. 10794). – (143v) Oracio. Domine Ihesu Christe fili Dei vivi qui dixisti nolo mortem peccatoris sed ut magis convertatur et vivat queso pone amaram mortem … (143v) Prim. Hymnus (AH 32, Nr. 130). – (144r) Ps 117. – Ps 120. – (144v) Ps 121. – (145r) Ps 122. – (145v) Antiphon (CAO Nr. 3707). – Capitulum. – (146r) Responsorium mit Versikel. – Oracio. Deus qui salutis eterne beate Marie virginitate fecunda humano genere premia prestitis ti … – (146v) Oracio. Domine Ihesu Criste fili Dei vivi qui dixisti … (identisch mit Bl. 143v). Cod. 353 261 (147r) Terz. Hymnus (AH 32, Nr. 130). – Ps 123. – (147v) Ps 124. – (148r) Ps 125. – Antiphon (CAO Nr. 1570). – Capitulum. – (149r) Oracio. Concede misericors Deus fragilitati nostre pre sidium ut qui sancte Dei genitricis Marie memoriam … – Oracio. De omnibus sanctis. Deus qui nos concedis omnium sanctorum memoriam agere … – (149v) Oracio. Domine Ihesu Criste fili Dei vivi qui dixisti … (identisch mit Bl. 143v). (149v) Sext. Hymnus (AH 32, Nr. 130). – (150r) Ps 126. – (150v) Ps 127. – (151r) Ps 128. – Anti phon (CAO Nr. 4699). – (151v) Capitulum. – Responsorium mit Versikel. – Oracio. Famulorum tuorum quesumus domine delictis ignosce ut qui tibi placere … – (152r) Oracio. Domine Ihesu Christe fili Dei vivi qui dixisti … (identisch mit Bl. 143v). (152r) Non. Hymnus (AH 32, Nr. 130). – (152v) Ps 129. – (153r) Ps 130. – (153v) Ps 131. – (153Ir) Antiphon (CAO Nr. 5040). – Capitulum. – Responsorium mit Versikel. – Oracio. Porrige nobis Deus dexteram tuam et per intercessionem sanctissimi Dei genitricis Marie auxilium nobis superne virtutis impende … – (153Iv) Oracio. Domine Ihesu Criste fili Dei vivi qui dixisti … (identisch mit Bl. 143v). (153v) Vesper. Ps 110. – (154v) Antiphon (CAO Nr. 2547). – Ps 113. – (155r) Antiphon (CAO Nr. 4937). – Ps 122. – (155v) Antiphon (CAO Nr. 2855). – Ps 127. – (156r) Antiphon (CAO Nr. 2641). – Ps 147,12‑20. – (156v) Antiphon (CAO Nr. 2887). – Capitulum. – Responsorium mit Versikel. – (157r) Hymnus (AH 2, Nr. 29). – (157v) Magnificat (nur Verweis). – Antiphon (CAO Nr. 1542). – Oracio. Supplicacionem servorum tuorum Deus miserator exaudi ut qui in commemoracione sancte Dei genitricis et virginis Marie congregamur eius intercessionibus … – (158r) Oracio. Domine Ihesu Christe fili Dei vivi qui dixisti … (identisch mit Bl. 143v). (158r) Komplet. – (158v) Ps 132. – (159v) Ps 133. – Ps 134. – (160r) Antiphon (CAO Nr. 5040). – Capitulum. – Responsorium mit Versikel. – (160v) Hymnus (Chev. Nr. 6346). – (161r) Canticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 2952). – Oracio. Quesumus omnipotens Deus ut beate et gloriose semperque virginis Dei genitricis Marie intercessio gloriosa nos ab omni malo protegat … – (161v) Conclusio. Has horas canonicas Maria mater pia tue laudi refero cordis simphonia … – Oracio etc. Ave gloriosa Maria mater Cristi virgo pia pone misericordiam et compassionem … – (162r) Oracio devotissima de compassione gloriose virginis Marie etc. Sta bat mater dolorosa iuxta crucem lacrimosa dum pendebat filius … – (163r) Oracio. Interveniat pro nobis domine Ihesu Criste nunc et in hora nostre mortis apud tuam clementiam gloriosa Dei genitrix virgo Maria … (163v) ganzseitige Miniatur (s. bei A). 11 (165r–183r) Cursus de septem gaudiis Mariae virginis. Hymnen (AH 30, Nr. 52). Tit.: Cursus de septem gaudiis Marie virginis. (165r) Invitatorium mit Verweis auf Ps. 95. – Matutin. Hymnus. – (165v) Ps 43. – (166r) Anti phona. Gaude celum trinitatis celans Dei filium planta virens puritatis iocundum ferens lilium tu regina virginum tu lux plena luminum … – Benediccio. Precibus et meritis beate Marie virginis liberet nos filius eius ab omnibus malis … – (166v) Leccio etc. Inc.: Gaude felix mater cum tibi per angelum Dei filius nuncciatur cum pudicus alvus tuus spiritu sancto fecundatur … – Expl.: … angeli leticiam iusti graciam et peccatores veniam inveniunt in eternum. Tu autem. – (167r) Responsorium. – (167v) Te Deum Marianum. (Chev. Nr. 20158. Walther, Initia Nr. 19052). (169v) Laudes. – (170r) Ps 117. – Antiphona. Gaude virgo virginum virga Yesse celeste ortans florem … – Capitulum. – (170v) Hymnus. – (171r) Canticum Zachariae. – (171v) Antiphona. Gaude virgo stirps Davitica flos fecundus fructus iocunditatis … – (172r) Oracio. Letifica et adiuva qui sumus omnipotens et misericors Deus dominice anuncciacionis et concepcionis gau dia recolentes … (172v) Prim. Hymnus. – (173r) Ps 87. – (173v) Antiphona. Gaudet celum letantur sidera caste matris exultant viscera … – Capitulum. – Responsorium mit Versikel. – (174r) Oracio. In men 262 Cod. 353 tibus nostris quesumus domine tue gratie lumen infunde ut qui conceptum et natum de virgine Deum verum et hominem confitemur … (174r) Terz. – (174v) Hymnus. – (175r) Psalmus (recte: Ies. 12). – (175v) Antiphona. Gaudet mater et filia novo regis preconio plaudit celorum curia celesti nato parvulo … – Capitulum. – Responsorium mit Versikel. – (176r) Oracio. Deus qui novo stelle signo novum filium regibus ostendisti presta quesumus ut intercedente benedicta et gloriosa semper virgine … (176r) Sext. Hymnus. – (176v) Ps 18,47–50. – (177r) Antiphon (CAO Nr. 4597). – Capitulum. – Responsorium mit Versikel. – Oracio. O domine Ihesu Criste fili Dei vivi benedicti qui pro nobis peccatoribus agonisans in cruce predilectam matrem … (177v) Non. Hymnus. – (178r) Ps 68,33–36. – Antiphona. O regina supernorum summo gaude gaudio psallat chorus angelorum cum regis fit ascensio dulcis cantus … – (178v) Capitulum. – Responsorium mit Versikel. – Oracio. Festina quesumus domine Ihesu festina ne tardaveris et auxilium nobis superne virtutis impende … (179r) Vesper. Ps 51,12–16. – (179v) Antiphona. Ihesu bone permissum spiritum peccatori di mitte debitum pie matris attendens meritum dirum … – Capitulum. – Hymnus. (180v) Magnificat (nur Verweis). – Antiphona. Clemens virgo mater pia cor inflamma frigidisisque nobis recta via … – Oracio. Deus qui benedicte et gloriose semper virginis genetricis filii tui Marie corpus et animam sancti spiritus illustracionem perfecisti … (180v) Komplet. Ps 45,10–14. – (181r) Antiphona. O decus innocencie flos virginitatis mater regis gloriose templum trinitatis regina clemencie … – Capitulum. – (181v) Responsorium mit Versikel. – Hymnus. – (182r) Canticum Simeonis (nur Verweis). – Antiphona. O mater egregia tutrix orphanorum portus spes fiducia facta peccatorum … – (182v) Oracio. Deus qui beatissi mam semper virginem Mariam in conceptu et partu virginitate servata triplici gaudio … – Se quitur quedam prosa de omnibus septem gaudiis beate virginis Marie. Gaude virgo que de celis iuxta vocem Gabrielis concepisti filium. Gaude mater Ihesu Christi quia virgo peperisti creato rem omnium … (183v) ganzseitige Miniatur (s. bei A). 12 (184r–195v) Cursus de anima Christi, unvollständig. Hymnen (AH 30, Nr. 8). Tit.: Cursus de anima Christi ut sequitur. (184r) Invitatorium mit Verweis auf Ps. 95. – Matutin. Hymnus. – (184v) Ps 34,2–6. – Antiphon (CAO Nr. 4418). – (185r) Benedictio. Exaudi domine preces servi tui ad te clamantis qui in trinitate perfecta vivis et regnas Deus … – Leccio. Inc.: Domini Ihesu felix anima divinitati personaliter unita humane infirmitatis defectus habuit mihi tristis fuit … – Expl.: … et in eius lucem splendoremque tenens cum ipso principaliter unus (!) spiritus est facta. Tu autem. – (185v) Responsorium mit Versikel. – Te Deum. (185v) Laudes. Ps 63,6–12. – (186r) Antiphon (CAO Nr. 3261). – (186v) Capitulum. – Hymnus. – (187r) Canticum Zachariae (nur Verweis). – Antiphon (CAO Nr. 5454). – Oracio. Celesti quesumus domine lumine semper et ubique nos preveni rationalis … (187v) Prim. Hymnus. – Ps 30,2–6. – (188r) Antiphon (CAO Nr. 2547). – Capitulum. – Respon sorium mit Versikel. – (188v) Oracio. Illumina quesumus domine populum tuum et splendore passionum carismatum et glorie racionalis ac beatissime anime unigeniti filii tui domini nostri Ihesu Cristi … (188v) Terz. Hymnus. – (189r) Ps 86,1–6. – (189v) Antiphon (CAO Nr. 2855). – Capitulum. – Responsorium mit Versikel. – Oracio. Presta quesumus omnipotens Deus ut passionum carisma tum et glorie racionalis ac beatifice anime unigeniti filii tui domini nostri Ihesu Cristi … (190r) Sext. Hymnus. – Ps 119,81–84. – (190v) Antiphon (CAO Nr. 4343). – Capitulum. – Res ponsorium mit Versikel. – Oracio. Domine Ihesu Criste qui dum hora sexta pro redempcione mundi crucis ascendisses lignum … Cod. 360 263 (191r) Non. Hymnus. – (191v) Ps 31,6–9. – Antiphon (CAO Nr. 2641). – Capitulum. – (192r) Responsorium mit Versikel. – Oracio. Accepta sit quesumus domine in conspectu tuo nostra devocio ut passionum carismatum et glorie rationalis ac beatissime anime … (192r) Vesper. Ps 84,2–5. – (192v) Antiphon (CAO Nr. 2887). – Capitulum. – Responsorium mit Versikel. – (193r) Hymnus. – (193v) Magnificat (nur Verweis). – Antiphon (CAO Nr. 5162). – Oracio. Domine Deus virtutum qui collapsa restauras et restaurata conservas … (194r) Komplet. Ps 16,7–11. – (194v) Antiphon (CAO Nr. 5329). – Capitulum. – (195r) Respon sorium mit Versikel. – Hymnus. – (195v) Canticum Simeonis. – Antiphon (CAO Nr. 5325). – Oracio [ U. S. Cod. 360 CONRADUS HIRSAUGIENSIS Perg. I, 26, I Bl. 165 × 110. Entstehungsort unbestimmt, 12. Jh. B: Perg. tw. stark fleckig, vereinzelt kleine Löcher. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 3.IV24 + I26 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufge klebt. Letzte Lage möglicherweise urspr. ebenso Quaternio, in dessen Lagenmitte Blätter herausgenommen wurden (s. bei Inhalt). Am oberen Blattrand Foliierung 19 Jh. durch die ULB Tirol. S: Schriftraum 135 × 70/80, von kaum sichtbaren Bleistiftlinien gerahmt, zu 22 (Bl. 1r–8r, Schreiber 1–3) und 32 (Bl. 8v–25v, Schreiber 4–9) Zeilen auf kaum sichtbarer Bleistiftlinierung. Zirkelstiche, zumeist beschnit ten. Karolingisch-gotische Mischschrift von mehreren Händen: 1) Bl. 1r, Z. 1–3; 2) Bl. 1r, Z. 4–Bl. 7v, Z. 13; 3) Bl. 7v, Z. 14–Bl. 8r; 4) Bl. 8v–10r, Z. 8; 5) Bl. 10r, Z. 9–Bl. 14r, Z. 13; 6) Bl. 14r, Z. 14–Bl. 16v; 7) Bl. 17r; 8) Bl. 17v–24v; 9) Bl. 25r–v; 10) Bl. 26r–v. Bl. 17v–24r Marginalbuchstaben zur Kennzeichnung der Sprechereinsätze. Vereinzelte Marginal- (tw. beschnitten) und Interlinearglossen. A: Bl. 1r–7v rote ein- bis dreizeilige Lombarden, Bl. 7v mit einfachem Dekor im Binnenfeld. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im zweiten Feld Golddruck M. S. VAR. MAT. in Goldrahmung, im dritten Signatur schild (s. bei G). Schnitt rot gefärbt. Spiegel Pap. 18. Jh. Am Spiegel des VD Signaturen und Spuren einer Bleistiftnotiz (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Auf hebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 F 2 (?) (Tinte, radiert), II1 E 4 S. 2 (Bleistift) und 360 (Tinte) sowie Spuren einer Bleistiftnotiz. Am Rücken gelbes Signaturschild der ULB Tirol 360. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. Ir Stempel der Soprintendenza Tridentina 20. Jh. L: P. Lehmann, Literaturgeschichte im Mittelalter I. Germanisch-Romanische Monatsschrift 4 (1912) 572. – R. B. C. Huygens, Conrad de Hirsau, Dialogus super auctores. Latomus 17 (1955) 5. – K. Langosch, Über lieferungsgeschichte der mittellateinischen Literatur, in: Geschichte der Textüberlieferung II. Zürich 1964, 184 Anm. 351. – R. B. C. Huygens, Accessus ad auctores. Bernard d’Utrecht. Conrad d’Hirsau, Dialogus super auctores. Édition critique entièrement revue et augmentée. Leiden 1970, 10. – L. G. Whitbread, Con rad von Hirsau as literary critic. Speculum 47 (1972) 234–245, 235. – Neuhauser, Neustift 84. – V. Brown, A twelfth-century Virgilian miscellany-commentary of German origin (Vatican, Ms. Pal. Lat. 1695), in: S. Krämer, M. Bernhard, Scire litteras. Forschungen zum mittelalterlichen Geistesleben (Abh. der Bayerischen Akademie der Wissenschaften, phil.-hist. Kl. N. F. 99). München 1988, 73–86, hier 74. – Katalog ULB Tirol IV 252, 472. – R. Marchionni, Dialogo sugli autori. Corrado di Hirsau. Introduzione, testo, traduzione e note di commento (Annali dell’Università di Napoli „L’Orientale“ [AION]. Dipartimento di studi del mondo classico e del mediterraneo antico. Sezione filologico-letteraria, Quaderni 12). Pisa, Roma 2008, insbes. 30–31. 264 1 Cod. 397 (Ir–v) leer bis auf Stempel (s. bei G). (1r–25v) ‹Conradus Hirsaugiensis›: Dialogus super auctores, unvollständig (Ed. Huygens 1970, 71–131 [basierend auf Neustift, Stiftsbibl., Cod. 360 und Würzburg, UB, M. p. th. f. 53]). Inc.: Quia video te diu vacare ocio silentio … – Expl. mut. (V. 1506): … in quorum ponderoso sensu et litera proprietate magna prolata [ Zur Textüberlieferung: Huygens 1970, 10–17. 2 (26r) ‹Ps.-Beda Venerabilis›: Termini quartae decimae lunae regulares (Memorialverse zur Be rechnung des Osterfestes) (MGH Poetae IV 670–671. PL 90, 708B und 802A). Inc.: None Aprilis norunt quinos … Lit.: B. Bischoff, Ostertagtexte und Intervalltafeln, in: Ders., Mittelalterliche Studien. Aus gewählte Aufsätze zur Schriftkunde und Literaturgeschichte II. Stuttgart 1967, 192–227, hier 192. 3 (26r) Notizen: 1) durchgestrichene und radierte Notiz (unleserlich). 2) verblasst: Isti sunt filii […] Iohannes bonus et Lafrancus filius Ambrosii est Albertus Zi cerius … 4 (26v) Tafel zur Berechnung des Osterfestes mit erläuternden Versus de termino paschae (19 Zeilen) (D. Schaller, E. Könsgen, Initia carminum Latinorum saeculo undecimo antiquiorum. Göttingen 1977, Nr. 208. A. Cordoliani, Contribution à la littérature du comput ecclésiastique au moyen âge. Studi medievali Ser. 3, 1/1 [1960] 112 und 2/1 [1961] 182. Walther, Initia Nr. 444). Am oberen Blattrand Bezeichnung der Spalten der Tabelle. Inc. Versus: Ad XVI numerum quintum dato primum … – Expl.: … Tandem cum binis XXVI dato trinis. Lit.: B. Bischoff, Ostertagtexte und Intervalltafeln, in: Ders., Mittelalterliche Studien. Aus gewählte Aufsätze zur Schriftkunde und Literaturgeschichte II. Stuttgart 1967, 192–227. Darunter komputistische Notizen. (Ir–v) leer. P. K. Cod. 397 IOHANNES DE TURRECREMATA Perg. I, 88, I Bl. 140 × 105. Süddeutscher Raum (?), 2. H. 15. Jh. B: Dickes Perg. Lagen: (I–1)I (Vorsatzbl.) + 11.IV88 + (IV–7)I* (Nachsatzbl.). Gegenbl. zum Vorsatzbl. als Spie gel auf dem VD aufgeklebt. Sechs Blätter der letzten Lage herausgeschnitten: Gegenbl. zum herausgeschnit tenen zweiten Blatt erhalten (= Nachsatzbl.), Gegenbl. zum herausgeschnittenen ersten Blatt als Spiegel auf dem HD aufgeklebt. In der ersten Lagenhälfte jeweils Kustoden, beschnitten. Am oberen Blattrand Foliie rung 19. Jh. durch die ULB Tirol. S: Schriftraum 90 × 65, von feinen, kaum sichtbaren Tintenlinien seitlich gerahmt, zu 14 Zeilen auf feiner Tin tenlinierung. Textualis. A: Rote Überschriften und Schlussschrift. In der ersten Zeile Oberlängen von Buchstaben bisweilen mit cadel lenartigen Ausläufern. Platz für zwei- bzw. dreizeilige Lombarden freigelassen, die später (Barockzeit?) nachgetragen wurden (z. B. M Bl. 34v). Repräsentanten. E: Gotischer Einband: braunes Leder über Holz mit Blinddruck. Süddeutscher Raum, 2. H. 15. Jh. Cod. 397 265 Leder abgerieben und tw. beschädigt, insbes. an den Ecken, am Rücken oben und unten eingerissen. VD: zwei von jeweils dreifachen Streicheisenlinien gebildete Rahmen. Rahmen 1: Rolle mit Vase, Wappenschild und Engel, tw. stark abgerieben (Nr. 1); Rahmen 2: Einzelstempel Schriftband „Maria“ (Nr. 2), die Ecken der Rahmenbegrenzungen durch dreifache Streicheisenlinien miteinander verbunden. Im Mittelfeld Rolle zwei Quadrate konkav nebeneinander, gefüllt mit Fünfpunktblüte (Nr. 3). HD: zwei von jeweils dreifachen Streicheisenlinien gebildete Rahmen. Rahmen 1: Rolle mit ornamentalem Blattwerk in geometrischer An ordnung (Nr. 4); Rahmen 2 (innere Begrenzungslinien bis an die äußeren verlängert): seitlich je ein Schrift band „Maria“ (Nr. 2), oben und unten leer. Im Mittelfeld Rolle (Nr. 1). Zwei Leder-Metall-Schließen. Kan ten gerade. Rücken: drei Doppelbünde. Im ersten Feld Papierschild M. S. (Tinte), im zweiten Papiertitelschild Turrecremata Meditatio[…] (Tinte), im dritten Signaturschild (s. bei G), im vierten leeres Papierschild. Kapitale mit grün-rosa Garn umstochen. Spiegel Perg. Am Spiegel des VD Exlibris, Signaturen (s. bei G) sowie Notiz (s. bei Inhalt), am Spiegel des HD Gebet und Federproben (s. bei Inhalt). G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Bl. 1r Besitzvermerk 18. Jh. Collegii Nova cellensis. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II1 F 18 (Bleistift) und 397 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 397. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. Ir Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller I 439. – C. F. Bühler, Rezension zu Kristeller I, Speculum. A Journal of Mediaeval Studies 39 (1964) 712–713, hier 712 (dazu J. N. Garvin, Scriptorium 19 [1965] 307, B 545). – Kaeppeli Nr. 2736. – Katalog ULB Tirol IV 446, 472. (VDS) Notiz. Si vixeris secundum carnem / morieris morte aeterna. (Ir–v) leer bis auf Stempel (s. bei G). (1r–88v) Iohannes de Turrecremata: Meditationes (Druck: u. a. GW M48257. Faksimile-Ausgabe des Erstdrucks von 1467 nach dem Exemplar der Stadtbibliothek Nürnberg, ed. H. Zirnbauer. Wiesbaden 1968. – Kaeppeli Nr. 2736, mit Nennung der Hs.). (1r) Tit.: Meditationes reverendissimi patris domini Iohannis de Turrecremata sacrosancte Romane ecclesie cardinalis posite et depicte de ipsius mandato in ecclesie ambitu sancte Marie de Minerva Rome. (1r) Tit.: De Dei potentia sapiencia ac benignitate. Meditatio prima. – Inc.: O admiranda et laudanda tue dispensationis gratia omnipotentissime domine Deus. Quia enim in natura tua ab homine videri non poteras ut hominem noticie tue participem faceres opus fecisti … – Expl.: … continue habere re commissos. (88v) Schlussschrift: Finite sunt contemplaciones reverendissimi patris domini Iohannis de Turre crema. Deo gracias. (Ir) Federprobe: Pater noster qui es in coelis (übermalt). (Iv–HDS) Mariengebet, dt. Tit. verblasst. Inc.: Maria du hochgelobter Namen ein Khönigin des Himels … – Expl.: … embpfangen. Darunter Federproben. C. S. 266 Cod. 405 Cod. 405 BREVIARIUM SALISBURGENSE Perg. VII, 236 Bl. 135 × 100. Wohl Neustift, um 1500 (u. a. 1501). B: Hs. im 20. Jh. neu gebunden (sehr eng), hierbei zahlreiche Fälze von sehr starrem Perg. eingefügt, daher ein zelne Blätter heute schadhaft und beinahe gelöst (kein Restaurierbericht erhalten). Lagen: (I–1)Ia + (IV–2)VII + 2.V20 + IV28 + V38 + IV46 + 8.V126 + 2.IV142 + 9.V221(232) + (V–6)225(236). Gegenbl. zu Bl. Ia als Spiegel auf dem VD aufgeklebt, heute weitgehend gelöst. Gegenbl. zu Bl. I und II herausgeschnitten (ohne Textverlust). Gegenbl. zu Bl. 222 als Spiegel auf dem HD aufgeklebt, Gegenbl. zu Bl. 222–225 sowie ein Doppelbl. herausgeschnitten (jeweils ohne Textverlust). Um Bl. 1 und 10, Bl. 88 und 95, Bl. 130 und 131, Bl. 136 und 141, Bl. 183 und 190, Bl. 195 und 199, Bl. 205 und 208, Bl. 213 und 220, Bl. 216 und 217 jeweils schmaler Falz geschlagen. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: auf 177 folgt 167, daher 167–177 doppelt angeführt. Bl. 1, 106, 126, 150, 170 Spuren bzw. Reste von Signakeln. S: Schriftraum 105/110 × 75, von feinen Tintenlinien seitlich begrenzt, zu einheitlich 24 Zeilen auf Blind linierung. Bastarda. Heiligenlitanei Bl. 130r–131r: Zwei Spalten, von feinen Tintenlinien gerahmt. Kalendar Bl. Ir–VIv: Schriftraum 115 × 80, mit Tintenlinien in fünf Spalten unterteilt, Tintenlinierung. Bastarda von gleicher Hand. Bl. 225r–v Nachtrag Textualis 16. Jh. Bl. 225r Datierung 1501, unklar, ob auf Haupttext oder auf Nachtrag bezogen. A: Rote Auszeichnungsstriche (vereinzelt), Überschriften. Satzmajuskeln mit tw. gelängten Schäften und cadellenartigen Verzierungen. Ein- bis vierzeilige (dunkel-)rote und blaue Lombarden (als KL-Ligaturen ca. zweizeilig, in der Litanei Bl. 130r–131r S[ancte]-Schleifen) mit Punktverdickungen sowie fallweise mit kopfstempelartigem Besatz und Ausläufern, die in gestrichelte Achterschlingen (Bl. 214v und 161v Spiral ranken, Bl. 161r Flechtknoten) gelegt und u. a. mit freien Kreisen und Fibrillen dekoriert sein können; sehr ähnliche Lombardenausläufer in Cod. 85 und 194 (vgl. z. B. Bl. 160v mit Cod. 85, Bl. 1r bzw. Cod. 194, Bl. 22r); einige der Lombarden (v. a. Bl. 204v–222r, drei- und vierzeilig) mit Fleuronnée in der Gegenfarbe. Repräsentanten. Flüchtiges Fleuronnée; in den Binnenfeldern längliche, zuweilen (leicht) blasige Knospen in verschiedenen Formationen sowie gegenständige (halbpalmettenartige) Blattpaare, Bl. 124r mit tropfenförmiger Mittel ader, gefüllt mit kleinen Kreisen („Erbsenschotenmotiv“); Konturlinien; als Besatz u. a. Perlen und Knospen, charakteristisch eine Kombination aus zumeist drei Knospen (tw. bedornt), die in zwei kürzere Fäden aus laufen können; kurze, schwungvolle Ausläufer; Bl. 204v Profilgesicht mit wulstiger Stirn und Nase; zur Einordnung s. u. Deckfarbenschmuck: 24 drei- bis sechszeilige Ornamentalinitialen, gut die Hälfte mit (sehr) kurzen Ranken, v. a. die kleineren Initialen rankenlos; zwei Wappen; drei ganzseitige Miniaturen. Die ornamentalen Initialen (Bl. 1r, 2r, 14v, 19v, 31v, 41v, 51v, 66r, 77v, 92v, 106r, 119r, 136r, 163v, 169v, 169Ir, 176Ir, 185v, 199r, 206v, 208v, 216v, 211r, 221r) von unterschiedlicher Qualität, die kleineren Initialen bes. nachlässig gemalt (z. B. Bl. 14v), aber wohl alle von einer Hand. Quadratische Initialfelder mit plumpen Rahmen(linien). Buchstabenkörper gerne blau; weiße und v. a. (ocker-)gelbe Höhungen; einfache Blatt füllungen, u. a. mit kopfstempelartigen Aussparungen (eigentlich nur Stempelknäufe); Bl. 77v Stufenband. Binnenfelder (auch Außengründe) häufig blattvergoldet, teils punziert (mit Blüten gefüllte Rautengitter), teils mit flüchtigem, gelblichem (Linien-)Dekor bemalt; in dieser Art auch der Buchstabenkörper Bl. 185v auf dem Goldgrund aufgebracht. Einige Buchstabenkörper gehen in kurze Ausläufer über; zudem recht derbe Akanthusranken (mit und ohne Kontakt zur Initiale), die unterschiedliche Blattformen zeigen (u. a. mit tropfenförmig verdickten Adern); Bl. 2r (und 106r) etwas aufwändigere Ranken mit Fantasieblüten; Bl. 216v separate Ranke mit naturalistisch gestaltetem Anschnitt; als Dekor Goldfelder (Bl. 106r in Eckschlinge) und -punkte mit rotem Dekor, u. a. freie Kreise und Fibrillen (vgl. Fleuronnée). Urspr. VDSv (100 × 66) Hl. Rochus mit Pilgerstab und -hut schlägt seinen Rock zurück, um Pestwunde am Oberschenkel zu zeigen; daneben an einem Baumstamm (?) der von Pfeilen durchbohrte hl. Sebastian (mit Lendentuch und erhobenem rechtem Arm). Beine des hl. Rochus tw. vom Grün des Wiesengrundes über deckt; Pfeile des hl. Sebastian abgeblättert oder abgekratzt. Bl. Iav (102 × 72) Engel (Dreiviertelfigur) präsentiert zwei Wappenschilde; links Propst Lukas Härber von Ringelsperg (Wappenschild geviert, Feld 1: auf rotem Grund goldenes Tau-Kreuz, Feld 2: auf blauem Grund drei diagonale Goldpfeile, Feld 3: auf blauem Grund goldener, gewundener Kranz, Feld 4: auf rotem Grund zwei überkreuzte goldene Schlüssel); rechts Neustifter Wappen (auf rotem, von zarten, gelblichen Ranken überzogenem Grund goldenes Tau-Kreuz); der Engel mit hohem Stirnkreuz und Flügeln, die über ihm einen Bogen andeuten, stützt die Wappen auf einer Steinbrüstung auf. HDS (104 × 68) Hl. Barbara mit Kelch (darüber Hostie, die abgeblättert zu sein scheint). Cod. 405 267 Das Fleuronnée von der im 1. Fünftel 16. Jh. nachweisbaren Hand, die auch in Cod. 194 (dat. 1507) und Cod. 142, T. I (dat. 1521) tätig war, in beiden Bänden des Zollner-Graduales einige Initialen mit Fleuronnée (s. jeweils Ergänzungshand 2) geschmückt und Sign. 14820 zur Gänze ausgestattet hat. Diese Hand ist ver mutlich auch für den ornamentalen Deckfarbendekor in der vorliegenden Hs. sowie in Cod. 194 verantwort lich; dieser Dekor und das Fleuronnée in Cod. 405 allerdings flüchtiger ausgeführt. Detailliert zu dieser Hand und den Zusammenhängen zw. Fleuronnée- und Deckfarbenzier s. bei Sign. 14820. Die Heiligenminiaturen stammen von einer Hand: bunte, breite Rahmungen; Goldgründe, auf denen mit gelblichem Malmittel Spiralranken und Heiligenscheine (bei hl. Katharina auch Krone) eingetragen sind; simple Bodengestaltungen; stimmige Figurenanatomie, ovale Gesichter mit hoher Stirnpartie, großflächigen Oberlidern und gerader Nase; Haarwellen der hl. Katharina mit breitem Pinsel in regelmäßigen Schwüngen gestaltet; Gewanddraperien mit einfachen, aber sicheren (Schraffur-)Linien definiert. – Wappenhalterengel Bl. Iav von anderer Hand: Rahmen in ockerfarbenem Pinselgold, Hintergrund kobaltblau (mit dünnlinigem weißem Muster), flankiert von breiteren Goldstreifen (punziert mit zwölfblättrigen Blüten); Engel mit paus bäckigem Gesicht, zwei unterschiedlich geschnittenen Augen und wolligem Haar; sein zart rosafarbenes Gewand mit routiniert ausgeführten Faltenbahnen, zudem gekonnte Schattenwürfe, die der Komposition Tiefe verleihen. Der Engel zeigt einige Ähnlichkeiten zur Miniatur des hl. Augustinus im 1507 geschaffenen Cod. 194; vgl. z. B. die Faltenbildung des Engelskleides und des Untergewands des hl. Augustinus; darüber hinaus stehen Engel und Augustinus hinter einer Fensterbrüstung; Ausführung der Miniaturen in Cod. 194 jedoch wesentlich aufwändiger und subtiler. S. R. E: Renaissance-Einband: braun gefärbtes Leder über Holz mit Blind- und Golddruck. Neustift, 2. H. 16. Jh. Werkstatt: Narziß Schuechl. Leder tw. leicht abgerieben, am VD Bünde tw. freiliegend, unteres und oberes Häubchen tw. eingerissen. Holz des VD und HD leicht gewölbt. VD und HD gleich: am äußersten Rand durch dreifache Streicheisen linien (tw. auf Kante) eingefasst. Durch ebensolche Linien Einkerbungen in der Mitte des oberen, unteren und seitlichen Randes betont. Ein durch je dreifache Streicheisenlinien begrenzter Rahmen: Rolle mit in stilisierten Blättern auslaufender Volute in Golddruck (Nr. 1 = Neuhauser, Rolleneinbände, R 24). Im Mittel feld mit zweifachen Goldlinien eingeschriebene Raute, in der Mitte rautenförmiger floraler Einzelstempel frei (Nr. 2), in den Ecken jeweils Einzelstempel gebündeltes Dreiblatt in Quadratform (Nr. 3). Am VD zu sätzlich Einzelstempel kleine fünfblättrige Blüte frei (Nr. 4). Spuren bzw. Rest von zwei Leder-MetallSchließen, beim Materialwechsel sich stark verjüngend, urspr. wohl gleiche Schließen wie Cod. 353 und 654. Kanten gerade und nach innen abgeschrägt, sowohl am Spiegel des VD als auch am HD durch einfache Streicheisenlinie eingefasst. Rücken: drei Doppelbünde, oben und unten je ein einfacher Bund, durch Streicheisenlinien begrenzt bzw. bei Doppelbünden unterteilt, Begrenzung der Bünde am VD und HD fort gesetzt, spitz aufeinander zulaufend. In den Feldern zw. den Doppelbünden jeweils Einzelstempel gebündel tes Dreiblatt in Quadratform (Nr. 3), in den Feldern zw. Einzel- und Doppelbund Einzelstempel florales Dekor in Rhombusform (Nr. 5). An den Längsseiten des Rückens jeweils acht Einzelstempel achtblättrige Blüte frei (Nr. 6). Am oberen und unteren Rücken jeweils Rest eines aufgeklebten Papiertitelschilds, oben Psalte[rium] zu lesen. Unterhalb des ersten Doppelbundes Signaturschild (s. bei G). Kapitale mit gelbem und blauem bzw. tw. grünem Zwirn umstochen (Neubindung). Schnitt golden gefärbt, reich punziert mit pyramidal aufgebauten Kreisen, z. T. mit Binnenkreisen, und floralem Dekor. Urspr. Spiegel des VD heute größtenteils gelöst (s. bei Inhalt), auf dem Recto deutliche Klebespuren, heute Stempel, auf dem Verso Miniatur in Deckfarbenmalerei (s. bei A) sowie Signaturen (s. bei G). Unterhalb des urspr. Spiegels des VD beschriebenes Papierblatt aufgeklebt (s. bei Inhalt), darüber Exlibris (s. bei G). Am Spiegel des HD Miniatur in Deckfarbenmalerei (s. bei A). G: Aufgrund der Wappendarstellung auf Bl. Iav Hs. wohl im Auftrag von bzw. für Propst Lukas Härber von Ringelsperg (Propst 1483–1503) um 1500 entstanden, möglicherweise in Zusammenhang mit dem hand schriftlichen Adligat von ULB Tirol, 156 F 20 (s. bei Inhalt). Auch die Suffragien auf Bl. 216r–223r weisen auf Propst Lukas Härber hin (später zusätzlich Propst des Augustiner Chorherrenstiftes Waldsee, vgl. P. A. Beck, Lukas Härber, Probst von Waldsee und Neustift: ein Beitrag zur Geschichte des Chorherrenstiftes Waldsee. Diözesan-Archiv von Schwaben 18 [1900] 97–101). Bl. Iar Besitzvermerk 18. Jh. Collegii Neocell. Am heutigen Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Verso des urspr. Spiegels des VD alte Signaturen der ULB Tirol II1 F 6. S. 2 (Tinte, Bleistift) und 405 (Tinte), am Recto des urspr. Spiegels des VD Stempel der ULB Tirol 19. Jh. Am Rücken gelbes Signaturschild der ULB Tirol 405. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Am Recto des urspr. Spiegels des VD Stempel der Soprintendenza Tridentina 20. Jh. 268 L: Cod. 405 Wilhelm II 110. – Laußermayer 310. – Chev. II Nr. 14718 (ohne Nennung der Sign.) – AH 4, Nr. 397 und 398; AH 23, Nr. 139; AH 51, Nr. 107. – Neuhauser, Neustift 85. – Katalog ULB Tirol V 84, 673. – Stefani 107. – Kustatscher–Korenjak 158. – Baroffio 324, Nr. 16719–20. (VDS) Fragment aus einem philosophischen Traktat. Pap. 130 × 85, quer zum Buchblock eingeklebt. Erhaltener Schriftraum 85 × 100. Von feinen Tintenlinien seitlich begrenzt, 20 Zeilen erhalten, von Exlibris der Stiftsbibl. Neustift überklebt. Humanistische Minuskel 16. Jh. Inc. mut.: ] Item agitur scilicet qui […] non est generacione […] non est … – Expl. mut.: … super totam preposicionem […] so[…] videt non hominem et ut sic est communis (?) […] solum feratur super [ (urspr. VDSr) Stempel (s. bei G). (urspr. VDSv) Signaturen (s. bei G). Ganzseitige Miniatur (s. bei A). (Iar) leer bis auf Besitzvermerk (s. bei G). (Iav) ganzseitige Miniatur (s. bei A). (Ir–225v) Breviarium Salisburgense. Mit Breviarium Salisburgense (Druck: Venedig 1482 = GW 5442 = ULB Tirol, Ink. 156 F 20 [Kalendar mit handschriftlichen, auf Neustift bezogenen Ergänzungen]) weitgehend übereinstimmend, Anord nung abweichend. Die der Innsbrucker Inkunabel beigebundene Handschrift weist nach Neustift (vgl. insbes. Bl. 9v–10v); nicht zuletzt die übereinstimmende Reihenfolge der Suffragien sowie die Datie rung (vgl. jeweils Cod. 405, Bl. 221r–v und ULB Tirol, Ink. 156 F 20, Bl. 10v) erlauben die Annahme, dass Cod. 405 in Zusammenhang mit der Inkunabel samt Handschrift steht (vgl. auch Katalog X 469– 472); möglicherweise ist der handschriftliche Nachtrag in ULB Tirol, Ink. 156 F 20 mit Datierung von 1501 (Verbrüderung der beiden Augustiner Chorherrenstifte Waldsee und Neustift) auch erst zugleich mit Cod. 405 entstanden. In Cod. 405 weisen zudem Suffragien nach Stift Waldsee (s. bei G). 1 (Ir–VIv) Kalendar. Enthält in drei Spalten den Römischen Kalender, die Tagesbuchstaben und die Festbezeichnungen. Zudem Nennung der Anzahl der Lesungen, der Festgrade und Tierkreiszeichen. Die Hervorhebung in Rot von Ingenuin und Albuin, Kassian sowie der Dedicatio ecclesiae Brixi nensis verweisen auf die Diözese Brixen. 2 (1r–124v) Psalterium feriatum. Psalterium mit Antiphonen, Versikel, Capitula, Orationen und Benediktionen, gegliedert nach Tagzeiten sowie nach Wochentagen bzw. dies dominici und feriales unterschieden. Zudem jeweils unterschieden zw. Sommer- und Winterteil. (1r) Tit.: Incipit psalterium secundum usum ecclesie Salczburgensis. (1r–92r) Matutin und Laudes. (1r) Dominicis diebus. – (19v) Fer. 2. – (31r) Fer. 3. – (41r) Fer. 4. – (51r) Fer. 5. – (66r) Fer. 6. – (77v) Sabbatum. (92v) Prim. – (100r) Terz. – (102r) Sext. – (104r) Non. – (106r) Vesper. – (122r) Komplet. 3 (124r–129v) Preces ad omnes horas. (124r) Tit.: Sequuntur preces ad omnes horas per totum annum. (124r–125v) Matutin. – (124r) Ad matutinas in Quadragesima preces. Ps 51 (nur Verweis). (125r) Rubrik: Oracio ferialis vel de tempore. Finita oracione ad omnes horas dicatur super populum. Ps 51 (nur Verweis). Oracio. Omnipotens sempiterne Deus mestorum consolacio laborancium fortitudo … Ps 6, 120, 121 (jeweils nur Verweise). (125v–129r) Prim. – (125v) Ad primas in Quadragesima preces. – (126r) Rubrik: Hic incipe preces quotidianas ad primam per circulum anni. Zunächst Preces für die hohen Feiertage, ab Bl. 127r für die Werktage: Rubrik: Hucusque ad summa festa et ad binos dicuntur preces pre tacte. Ad primam. – (127v) Rubrik: Conclude cum versiculo et collecta ut supra in matutinis. Cod. 405 269 Ps 32, 122, 123 (jeweils nur Verweis). – (127v) Fünf abschließende Orationes mit vorangehender Rubrik: Ad primas per totum annum oracio. 1) Domine Deus pater omnipotens qui nos ad principium huius diei pervenire fecisti … Mit Zusätzen: Ad capitulum dicatur. Iube domine benedicere. Eterna Dei sapiencia illuminare dignetur sensus corda et corpora nostra … 2) Sancta Dei genitrix perpetua virgo Maria cum omnibus sanctis et electis diei intercedere dignare … Mit Zusätzen: Ter dicatur Gloria Kyrie Pater noster. Ps 90,16–17. 3) Dirigere sanctificare et regere digneris domine sancte pater omnipotens eterne Deus … Mit Zusätzen: Ad festum et binos. Ps 117. 4) Actiones nostras quesumus domine aspirando preveni et adiuvando prosequere … Mit Zu sätzen: Ad IX et tres lecciones dicitur. Ps 130 (nur Verweis). Kyrie. Pater noster. 5) Rubrik: Ad IX lecciones oracio. – Absolve quesumus domine animam fratrum et sororum pa rentum et benefactorum nostrorum et omnium fidelium defunctorum … (128v) Rubrik: De sanctis trium leccionum et in pura feria precum dicuntur tres collecte: 1) Beati Petri apostoli tui quesumus domine intercessione nos protege et animas famulorum tuorum sacerdotum sanctorum tuorum iunge … 2) Deus venie largitor et humane salutis amator quesumus clemenciam tuam … 3) Fidelium Deus omnium conditor et redemptor animabus famulorum famularumque tuarum omnium fidelium … (129r) Rubrik: In die animarum, in adventu domini et in Quadragesima feriatis diebus dicantur hi quinque psalmi: Ps 5, 6, 116, 1–9, 116, 10–19, 130 (jeweils nur Verweis). (129r) Rubrik: In adventu dicantur tres collecte ut supra. In Quadragesima quinque ut supra: 1) Beati Petri ut supra. 2) Deus indulgenciarum domine da fidelibus tuis quorum anniversarium deposicionis diem et memoriam agimus … 3) Deus venie largitor ut supra. Deus cuius misericordie non est numerus suscipe propicius preces humilitatis nostre … 4) Fidelium Deus ut supra. (129r) Benedictio: Adiutorium nostrum in nomine domini … (129v) Terz, Sext und Non. Preces ad tercias, sextas et nonas. (129v) Vesper und Komplet. Ad vesperas preces ut ad matutinas. – Ad completorium preces. Zu nächst Preces für die hohen Feiertage, anschließend für die Werktage: (129v) Rubrik: Hucusque ad festa et binos ut supra in primis. Anschließend Oracio. Deus qui diem diem (!) discernis a nocte … (129v) Abschließende Rubrik: Nota per annum extra Quadragesimam dicantur ad horas preces. 4 (130ra–132v) Litania. (132v) Rubrik: In Quadragesima additur psalmus Levavi oculos meos in (= Ps 121) cum Gloria patri. 5 (132v–135r) Preces votivae. (132v) Preces. Oremus pro omni gradu ecclesie. Sacerdotes tui induantur iusticia … – (132v) Oratio: Pietate tua quesumus domine nostrorum absolve vincula peccatorum … (133r) Rubrik: In angariis dicantur he 14 collecte. 1) Pro omni gradu ecclesie oracio. Omnipotens sempiterne Deus respicere dignare super apos tolicam dignitatem … 2) Pro episcopo. Omnipotens sempiterne Deus qui facis mirabilia magna solus … 3) Pro regibus et principibus. Omnipotens sempiterne Deus qui celestia simul et terrena mode raris … 4) Pro congregacione. Defende quesumus domine Deus beata Maria semper virgine … 5) Pro concordia oracio. Deus largitor pacis et amator charitatis da famulis tuis … 270 Cod. 405 6) Pro iter agentibus. Deus qui diligentibus te misericordiam tuam semper impendis et a servien tibus tibi in nulla … 7) Pro inimicis oracio. Deus pacis charitatisque amator et custos da omnibus … 8) Pro penitentibus. Omnipotens et misericos (!) Deus qui omnem animam confitentem et peni tentem … 9) Pro elemosinariis. Deus qui post baptismi sacramentum secundam abolicionem peccatorum elemosinis … 10) Pro quacumque tribulacione. Omnipotens sempiterne Deus mestorum consolacio laboran cium fortitudo … 11) Pro familiaribus. Deus qui charitatis dona per graciam sancti spiritus cordibus fidelium infundis … 12) Pro temptacione cogitacionum. Deus cui omne cor patet et omnis voluntas loquitur et quem nullum latet secretum … 13) Pro peticione lacrimarum. Omnipotens mitissime Deus qui sicienti populo fontem … 14) Pro vivis et defunctis. Omnipotens sempiterne Deus qui vivorum dominaris simul et mortu orum omniumque … (135r) Magnificat (Luc 1,46–55). – (135v) Pater noster. Credo. 6 (135v–163r) Hymnar. (135v–148r) Temporale. Tit.: Sequuntur hymni per totum annum. Et primo de tempore. (148r–163r) Sanctorale. Mit Druck weitgehend übereinstimmend: in der Hs. De confessoribus Ingenuino et Albuino (AH 23, Nr. 139, mit Nennung der Hs.) samt Laudes (AH 51, Nr. 107, mit Nennung der Hs.) sowie De sancta Monica matre Augustini (AH 4, Nr. 397, mit Nennung der Hs.) samt Nokturn (AH 4, Nr. 398, mit Nennung der Hs.) und Laudes (AH 19, Nr. 391) ergänzt. Tit.: Sequuntur nunc hympni de sanctis. 7 (163v–198v) Commune sanctorum. Mit Druck weitgehend übereinstimmend, Hymnen im Druck im Gegensatz zur Hs. im Hymnar enthalten, Lektionen gegenüber Druck gekürzt. Tit.: Incipit commune sanctorum et primo apostolorum. (163v) In festis apostolorum. – (169v) De pluribus martiribus. – (169Ir) In festo unius martiris. – (176Ir) De confessore et pontifice. – (185v) De simplici confessore. – (189r) De pluribus confes soribus. – (191r) De virgine et martire. – (195r) De virgine non martire. – (197r) De pluribus virginibus et martiribus. 8 (198v–206v) Vigiliae mortuorum. (198v) Tit.: Sequuntur vigilie mortuorum. 9 (206v–207v) Quindecim Gradus. (206v) Tit.: Sequuntur XV gradus. – Ad primum nocturnum. Ps 120–124 (jeweils Verweis). An schließend Versikel und Oratio: Absolve domine quesumus animas famulorum famularumque tuarum ab omni vinculo delictorum … – (207r) Secundus nocturnus pro congregacione. Ps 125– 129 (jeweils Verweis). Anschließend Versikel und Oratio: Deus cui proprium est misereri semper et parcere suscipe deprecacionem … – Tercius nocturnus pro familiaribus. Ps 130–134 (jeweils Verweis). Anschließend Versikel und Oratio: Pretende domine fidelibus tuis dexteram celestis auxilii … 10 (207v–208v) Orationes de ss. trinitate. (207v) Tit.: Sequitur trina oracio. – Invitatorium mit Psalm Deus in adiutorium und Psalm Ad te domine levavi (jeweils nur Verweis). Cod. 405 271 (207v) Ad patrem. Domine Deus pater omnipotens qui consubstancialem et coeternum tibi ante omnia secula ineffabiliter … (207v) Ad filium. Domine Ihesu Christe fili Dei vivi qui es verus et omnipotens Deus splendor et imago patris vita eterna … (208r) Ad spiritum. Domine sancte spiritus qui coeternus coequalis et consubstancialis patri filioque existens ab eis inenarrabiliter … (208r) Graciarum actio ad trinitatem. Gracias tibi ago omnipotens Deus qui me dignatus es custodire in hac nocte … 11 (208v–216r) Cursus BMV. (208v) Tit.: Sequitur cursus beate Marie virginis. Bei jeder Tagzeit zudem Einschub De passione Christi. (208v) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus (AH 50, Nr. 72, Str. 1–5), an schließend: Maria mater gracie mater misericordie tu nos ab hoste protege in hora mortis susci pe … – (209r) Antiphonen und Psalmen (jeweils nur Verweise) je nach Tagen verschieden: In dominica die, 2a et 5a feriis: Antiphonen (CAO Nr. 1709, Nr. 4942, 1438); Ps 8, 19, 24. Tercia et sexta feria: Antiphonen (CAO Nr. 4987, Nr. 1282, 4936); Ps 45, 46, 87. Quarta feria et sabbato: Antiphonen (CAO 2924, 2217, 4332); Ps 96–98. (209v) Benediccio. Sancta Dei genitrix sit nobis auxiliatrix. – Leccio 1a. Inc.: O beata et gloriosa virgo Maria quis digne tibi valeat iura graciarum … – Expl.: … que solo tuo commercio recupe randi aditum invenit. Tu autem. – Benediccio. Nos cum prole pia benedicat virgo Maria. – Leccio 2a. Inc.: Accipe itaque quascumque exiles quascumque meritis tuis impares graciarum accio nes … – Expl.: … et reporta nobis antidotum reconciliacionis. Tu autem. – (210r) Benediccio. Ad gaudia supernorum perducat nos regina celorum. – Leccio 3a. Inc.: Sit per te excusabile quod per te gerimus … – Expl.: … eiusdem iudicis et redemptoris. Tu. (Zu den Lesungen vgl. Fulbertus Carnotensis: Sermo IX [PL 141, 338]). (210r) Laudes. Ps 93 (nur Verweis). – (210v) Antiphon (CAO Nr. 1503). – Capitulum. – Hymnus (AH 2, Nr. 27, Str. 5–7). – Ps Benedictus (nur Verweis). – Antiphon (CAO Nr. 5041). – (211r) Oracio. Deus qui de beate Marie virginis utero verbum tuum angelo annunciante carnem susci pere voluisti … – In adventu domini oracio. Adiuvet nos quesumus domine beate Marie semper virginis intercessio veneranda … (211r) De passione Christi hora matutina. Hymnus (AH 30, Nr. 13, Str. 1). – Oracio. Domine Ihesu Christe absolve me per sanctam captivitatem tuam ob omnibus viciis … (211v) Ablassgebet: Anima Christi. Quicumque sequentem oracionem dixerit habet tria milia criminalium et XX milia venialium dierum indulgenciam. (211v) Prim. Hymnus (AH 32, Nr. 130, Str. 1–2 und 5). – (212r) Ps 120–122 (jeweils nur Ver weis). – Antiphon (CAO Nr. 4591). – Capitulum. – Oracio. Deus qui salutis eterne beate Marie virginitate fecunda humano generi premia prestisti … (212r) De passione domini ad primas. Hymnus (AH 30, Nr. 13, Str. 2). – (212v) Oracio. Domine Ihesu Christe qui ante iudices ligatis manibus stare voluisti … (212v) Terz. Hymnus (nur Verweis: AH 32, Nr. 130). – Ps 123–125 (jeweils nur Verweise). – Antiphon (CAO Nr. 5040). – Capitulum. – Oracio. Concede nos famulos tuos quesumus domine Deus noster perpetua mentis et corporis sanitate gaudere … (213r) De passione domini ad tercias. Hymnus (AH 30, Nr. 13, Str. 3). – Oracio. Domine Ihesu Christe sanctifica me per spineam coronam et per sacra vulnera … (213r) Sext. Hymnus (nur Verweis: AH 32, Nr. 130). – Ps 126–128 (jeweils nur Verweise). – Antiphon (CAO Nr. 4705). – Capitulum. – (213v) Oracio. Omnipotens sempiterne Deus famulos tuos dextera potencie tue a cunctis protege periculis … 272 Cod. 405 (213v) De passione domini ad sextas. Hymnus (AH 30, Nr. 13, Str. 4 [leicht abweichend]). – Oracio. Domine Ihesu Christe rogo te per amaram extensionem omnium membrorum et nerv orum tuorum … (213v) Non. – (214r) Hymnus (nur Verweis: AH 32, Nr. 130). – Ps 129–131 (jeweils nur Verwei se). – Antiphon (CAO Nr. 1357). – Capitulum. – Oracio. Protege domine famulos tuos subsidiis pacis et beate Marie virginis patrociniis confidentes … (214r) De passione domini ad nonas. Hymnus (AH 30, Nr. 13, Str. 5). – Oracio. Domine Ihesu Christe qui propter me mortem pati voluisti … (214v) Vesper. Ave Maria. – Antiphon (CAO Nr. 2547). – Ps 110 (nur Verweis). – Antiphon (CAO Nr. 4937). – Ps 113 (nur Verweis). – Antiphon (CAO Nr. 2855). – Ps 122 (nur Verweis). – Antiphon (CAO Nr. 2641). – Ps 127 (nur Verweis). – Antiphon (CAO Nr. 2887). – Ps 147,12 (nur Verweis). – Capitulum. – Ymnus. Ave maris (nur Verweis: De annunciacione supra in hymnario quere). – (215r) Antiphon (CAO Nr. 4703.3). – Oracio. Famulorum tuorum quesumus domine delictis ignosce ut qui tibi placere … (215r) De passione domini ad vesperas. Hymnus (AH 30, Nr. 13, Str. 6). – Oracio. Domine Ihesu Christe qui proprio sanguine tuo mundum redemisti redime me ab omnibus peccatis et negligen ciis … (215v) Komplet. Ps 132 (nur Verweis). – Antiphon (CAO Nr. 1862). – Hymnus (nur Verweis: AH 32, Nr. 130). – Capitulum. – Antiphon (CAO Nr. 2952). – Psalmus (recte: Lc 2,29–32). – Ora cio. Concede misericors Deus fragilitati nostre presidium ut qui sancte Dei genitricis Marie … (215v) De passione domini ad completorium. Hymnus (AH 30, Nr. 13, Str. 7). – (216r) Oracio. O dies eterne nunquam deficiens et sol nunquam occidens qui circa solis occasum sepultus suscepisti … 12 (216r–223r) Suffragia. 12.1 (216r–217v) Suffragia des Augustiner Chorherrenstiftes Waldsee. (216r) Tit.: Sequuntur suffragia contraria ad vesperas et ad matutinas secundum modum mo nasterii s. Petri in Walsee. Den einzelnen Gebeten der Suffragien jeweils eine Antiphon und ein Versikel vorangestellt. (216v) De s. trinitate. Omnipotens sempiterne Deus qui dedisti famulis tuis in confessione vere fidei eterne trinitatis gloriam agnoscere … – De sancta cruce. Deus qui crucem sanctam ascen disti et mundi tenebras illuminasti … – De beata virgine. Beate et gloriose semperque virginis Dei genitricis Marie quesumus omnipotens Deus intercessio gloriosa nos protegat … – (217r) De an gelis. Deus qui miro ordine angelorum ministeria hominumque dispensas concede propicius … – De s. Iohanne Baptista. Presta quesumus omnipotens Deus ut familia tua per viam salutis incedat et beati Iohannis precursoris … – De apostolis Petro et Paulo. Apostolicis nos quesumus domine beatorum apostolorum Petri et Pauli et aliorum apostolorum adtolle presidiis … – (217v) De s. Iohanne ewangelista. Ecclesiam tuam quesumus domine benignus illustra et (!) beati Iohannis apostoli tui et evangeliste illuminata doctrinis … – De s. Augustino. Adesto supplicacionibus nos tris quesumus omnipotens Deus et quibus fiduciam sperande pietatis indulges … – De omnibus sanctis. Da nobis domine continua omnium sanctorum tuorum veneracione letari … 12.2 (218r–220v) Suffragia ad placitum. (218r) Tit.: Sequuntur nunc alia suffragia ad placitum. Vor den einzelnen Orationen jeweils ein Versikel. (218r) De s. Maria virgine. Concede nos famulos tuos quesumus domine Deus noster perpetua mentis et corporis sanitate gaudere … – De s. Iohanne Baptista (nur Verweis). – De sancto Petro. Deus qui apostolo Petro collatis clavibus … – De apostolis Petro et Paulo. Protege domine po pulum tuum et apostolorum tuorum Petri et Pauli patrocinio confidentem … – De s. Augustino. Deus amator omnium bonorum qui sanctissimum Augustinum confessorem … – (218v) Alia ora cio. Adesto suplica[tionibus] (nur Verweis). – De s. Iohanne ewangelista. Ecclesiam tuam que Cod. 405 273 sumus … – De s. Iohanne Baptista. Concede quesumus omnipotens Deus ut quod beati Iohannis Baptiste solemnia colimus … – De s. Maria Magdalena. Deus qui beatam Mariam Magdalenam dominice resurreccionis gaudium sanctis apostolis innotescere voluisti … – De omnibus sanctis. Omnipotens sempiterne Deus qui nos omnium sanctorum tuorum merita sub una tribuisti celebritate … – Alia oracio. Da nobis quesumus domine continua omnium sanctorum tuorum veneracione letari … – (219r) De sancta Margaretha. Deus qui beatam virginem Margaretham ad celos per martirii palmam pervenire fecisti … – De s. spiritu. Deus qui corda fidelium sancti spiritus illustracione docuisti … – De angelis (nur Verweis). – De s. Katherina. Deus qui dedisti legem Moysi in summitate montis Synai et in eodem loco per sanctos angelos … – De s. Nicolao. Deus qui beatum Nicolaum pontificem tuum innumeris decorasti miraculis … – De s. cruce (nur Verweis) – De s. Dyonisio. Presta quesumus omnipotens Deus ut qui gloriosos martires Dyoni sium socios quam (!) eius fortes in sua confessione cognovimus … – (219v) De s. Laurencio. Da nobis quesumus omnipotens Deus viciorum nostrorum flammas extinguere … – De s. Andrea. Maiestatem tuam domine suppliciter exoramus ut sicut ecclesie tue beatus Andreas … – De s. Luca ewangelista. Interveniat pro nobis quesumus Deus sanctus tuus Lucas ewangelista qui crucis mortificacionem … – De corpore Christi. Deus qui nobis (gestrichen: per) sub sacramento mirabili passionis tue memoriam reliquisti tribue … – (220r) Pro pace. Deus a quo sancta desi deria recta consilia et iusta sunt opera … – Generalis oracio. A cunctis nos quesumus domine mentis et corporis defende periculis et intercedente gloriosa virgine Dei genitrice … – Oracio pro defunctis. Avete omnium fidelium anime in pace Christi requiescite quos redemit suo precioso sanguine … – (220v) Alia oracio pro defunctis. Sancte Deus sancte fortis sancte et immortalis Deus miserere animabus omnium fidelium defunctorum … – Alia oracio. Domine Ihesu Christe fili Dei vivi mundi creator et redemptor qui dixisti … 12.3 (221r–223r) Suffragia des Augustiner Chorherrenstiftes Neustift. (218r) Tit.: Sequuntur alia suffragia secundum chorum monasterii s. Marie in Novacella. Den einzelnen Orationen der Suffragien jeweils eine Antiphon und ein Versikel vorangestellt. Suffragien jeweils nach Tagen und Tagzeiten unterschieden. (221r) Et primo sabbatis noctibus ad vesperas. De sancta trinitate. Omnipotens sempiterne Deus qui dedisti famulis tuis in confessione vere fidei eterne trinitatis gloriam agnoscere … – De sanc ta cruce. Deus qui unigeniti filii tui precioso sanguine vivifice crucis vexillum sanctifficare (!) voluisti … – De beata virgine. Famulorum tuorum quesumus domine delictis ignosce ut qui tibi placere … – (221v) De sancto Augustino. Excita quesumus domine in ecclesia tua spiritum cui beatus Augustinus confessor tuus et pontifex servivit … – De omnibus sanctis. Maiestatem tuam domine supplices exoramus ut sicut nos iugiter omnium sanctorum … (221v) Dominicis diebus ad matutinas. De s. trinitate (nur Verweis). – De s. cruce. Adesto nobis domine Deus noster et quos sancte crucis letari fecisti … – (222r) De beata virgine. Protege domine famulos tuos subsidiis pacis et beate Marie virginis patrociniis confidentes … – De s. Augustino. Deus qui populo tuo beatum Augustinum eterne salutis ministrum constituisti … – De omnibus sanctis (nur Verweis). (222r) Per ebdomadam suffragia. Orationes durchwegs identisch mit obigen, lediglich Anti phonen und Versikel sind verändert. De beata virgine. – (222v) De sancto Augustino. – De omni bus sanctis. (223r) Suffragia ad matutinas per ebdomadam. Orationes durchwegs identisch mit obigen, ledig lich Antiphonen sind verändert. De beata virgine. – De sancto Augustino. – De omnibus sanctis. 13 (223v–224r) Accessus altaris (vgl. Jungmann I 354–355). Tit.: Sequitur accessus altaris. Ymnus. Veni creator (nur Verweis). – Ps 84–86, 116,10, 130 (jeweils nur Verweise). – Capitulum. – Preces. Ego dixi domine miserere mei. Sana animam meam quia … – Anschließend fünf Orationes: Aures tue pietatis mitissime Deus inclina precibus meis … – Deus cui omne cor patet et omnis voluntas loquitur … – (224r) Deus qui corda 274 Cod. 440 de[lium] (nur Beginn). – Auge in nobis domine fidem rectam spem firmam caritatemque fi perfectam … – Omnipotens sempiterne Deus dirige actus nostros in beneplacito tuo … 14 (224r–v) Zwei Orationes ante missam. Tit.: Sequitur ante missam oracio. 1) Pulcherrime Ihesu Christe rogo te per illam sacratissimam effusionem preciosissimi sanguinis tui quo sumus redempti … 2) Domine Ihesu Christe qui in cruce suspensus es pro nostra redempcione tuum dulcissimum spiritum in manus Dei patris … 15 (225r–v) Recessus altaris. Nachtrag (möglicherweise dat. 1501, s. bei S) (vgl. Jungmann II 572). (225r) Tit.: Sequitur recessus altaris. Canticum trium puerorum (Dn 3,57–88), Ps 150, Canticum Simeonis (Lc 2,29–32) (jeweils nur Textbeginn). Mit Antiphon. – Preces. Confiteantur tibi domi ne omnia opera tua … Exultabunt sancti in gloria … Non nobis domine non … Domine exaudi orationem. Anschließend drei Orationes: Deus qui tribus pueris mitigasti flammas ignium, con cede propicius … – Acciones nostras quesumus domine aspirando preveni et adiuvando prose quere … – (225v) Da nobis quesumus domine vitiorum nostrorum flammas extinguere … (HDS) ganzseitige Miniatur (s. bei A). U. S. Cod. 440 MARSILIUS DE INGHEN. IOHANNES BURIDANUS Pap. I, 414 Bl. 280 × 205. Tirol (?), um 1450. B: Pap. tw. fleckig, insbes. am Schnitt und ab Bl. 413. Bl. 13 lose. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 341, CCl 404, CCl 409. Lagen: II (Vorsatzbl.) + 7.VI84 + (VI–8)88 + 14.VI266(256) + (VI–10)268(258) + 13.VI424(414). Die letzten acht bzw. zehn Blätter der Lagen 8 bzw. 23 herausgeschnitten (ohne Textverlust). In den Lagenmitten tw. Pergamentfälze (bisweilen zweispaltig beschrieben und rubriziert). Lagenreklamanten, tw. be- bzw. weggeschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 143–152 ausgelassen. Bl. 88 und 268 neuzeitliche orange Papiersignakel der ULB Tirol. S: Schriftraum 195/225 × 145/155. Zwei Spalten, von Tinten- bzw. Bleistiftlinien gerahmt, zu 40–51 Zeilen. Zirkelstiche. Bastarda von drei Händen: 1) Bl. 1ra–88ra; 2) Bl. 89ra–268ra; 3) Bl. 269ra–423vb. Bei Schrei ber 1 und 3 Textbeginn in Textualis formata (Textura). A: Platz für Überschriften (bei Schreiber 2), Lombarden (bei Schreiber 3) und Initialen freigelassen, nicht aus geführt. E: Halbledereinband: weißes Schweinsleder über dickem Holz mit Blinddruck.Tirol (?), 2. H. 15. Jh. Lederüberzug am VD und HD etwa bis Mitte des Buchdeckels reichend. Leder am VD stark abgenützt und beschädigt. Am VD und HD von vierfachen Streicheisenlinien gebildeter Rahmen, Mittelfeld durch eben solche Linien diagonal unterteilt, Begrenzungsdreiecke gefüllt mit zahlreichen Einzelstempeln. Stempel am VD nicht mehr erkennbar, am HD: kleine Siebenpunktblüte (Nr. 1); Schriftband „Maria“ (Nr. 2); Doppelad ler mit Krone in Tropfenform (Nr. 3); Rosette mit zweifachem Blütenkranz frei (Nr. 4); Kielbogen mit Blüte frei (Nr. 5); nach links blickender Steinbock in Rhombus (Nr. 6). Zwei Leder-Metall-Schließen, verziert mit Schriftzug und achtblättriger Rosette (wie Neustift, Stiftsbibl., Cod. 4), oberes Schließband mit aufgekleb tem beschriebenem Papiertitelschild Marsilius. Kanten gerade. Rücken: blau gefärbt (wie öfters in Neustift), Fitzbünde, dazwischen drei Doppelbünde, Felder mit zweifachen Streicheisenlinien diagonal unterteilt. Im ersten Feld Papiertitelschild M: S: Marsilius de Passionibus, im zweiten Signaturschild (s. bei G), im vierten blau gefärbtes Papierschild (Reparatur?). Spiegel des VD und HD urspr. jeweils wohl Pergamentfragment, jetzt nicht mehr erhalten und nur noch Abklatsch sichtbar (liturgischer Text: Vigiliae Mortuorum, Lectio Cod. 440 275 VI–IX, Responsorien und Versikel; urspr. am VD aufgeklebtes Pergamentblatt heute gelöst; s. bei Inhalt), am VD zusätzlich Exlibris. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Die Wasserzeichen weisen in den Wiener Raum; verwandte Wasserzeichen u. a. auch in Innsbruck, ULB Tirol, Cod. 749, geschrieben in Schnals und Innichen. Bl. 1r Besitzvermerk 18. Jh. Collegii Neocell. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. Ir alte Signaturen der ULB Tirol II3 B 11 (Tinte, radiert), II2 B 12 (Bleistift) und 440 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 440 (mit Klebestreifen fixiert). Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Triden tina 20. Jh. L: Ch. H. Lohr, Medieval latin Aristotle commentaries. Traditio 27 (1971) 332. – M. Markowski, Katalog dzieł Marsyliusza z Inghen z ewidencją rękopisów (Katalog der Werke des Marsilius von Inghen mit der Evidenz der Handschriften). Studia Mediewistyczne 25, 2 (1988) 39–135, hier 108 und 122. – Kristeller I 439. – Ka talog ULB Tirol V 297, 673. (urspr. VDS, heute abgelöst) Fragment aus einem Breviarium (Brixinense?). Perg. ca. 277 × 210, urspr. als Spiegel auf dem VD aufgeklebt (Klebespuren am Verso), an den Blatträndern beschnitten. Schriftraum: 228 × 180, von Tintenlinien gerahmt (am Recto links von zweifachen Linien). Am Recto einspaltig, am Verso zwei Spalten, zu 28 Zeilen auf Tintenlinierung mit zahlreichen Ergänzungen am Blattrand. Textura 14. Jh., Ergänzungen großteils wohl von derselben Hand, vereinzelt von späterer Hand. Rote Auszeichnungs striche, Durchstreichungen, Paragraphzeichen und Überschriften. Rote ein- und zweizeilige Lombarden. Am oberen Blattrand links Signatur 440 (Tinte). 1) 2) 1 Recto: Enthält Gebete und Psalmhinweise aus dem Officium pro defunctis. Tit.: In deposicione alicuius defuncti. – Inc.: Commendo domine in misericordiam tue divinitatis maiestatis ac magnificencie … – Expl. mut.: … Sancte Stephane cum omnibus martiribus veni in adiutorium anime eius et animabus [ Am Blattrand Sequenzen. Am oberen Blattrand Inc. mut.: ] per quem videmus signa in populo. (rot) Sequencia sancte Marie. (AH 54, Nr. 237). Am linken Blattrand zwei Sequenzen (AH 54, Nr. 227; AH 32, Nr. 24), wobei die zweite am unteren Blattrand fortgesetzt wird. Verso (schlecht lesbar): Lectio VI–IX (Dn 12, 2–3; 1 Cor 15, 22–23; 1 Cor 15, 51–52; 1 Th 5, 2–5) mit Responsorien. Am oberen, rechten und unteren Blattrand Sequenzen (u. a. unten AH 55, Nr. 232). Die Lesungen entsprechen der Gruppe 9 bei Ottosen (K. Ottosen, The Responsories and Versicles of the Latin Office of the Dead. Aarhus 1993, 85–87), weit verbreitet im deutsch sprachigen Raum und auch in anderen Brixner Handschriften für die Vigiliae Minores belegt. Auch die Reihe der Responsorien (68–28–40–38) ist weit verbreitet; aus der Bamberger Tradi tion stammend lässt sie sich im gesamten deutschsprachigen Raum nachweisen, auch in Brixen. Hervorzuheben ist aber die Tatsache, dass das letzte Responsorium Libera me durchgestrichen und in Rot mit jenem aus den Vigiliae mortuorum sollemnes kommenden Deus aeterne (= Otto sen 18) ersetzt wurde. Dieses Responsorium begegnet in der einzig in Sign. 15063 überlieferten Responsorienreihe 70–44–47–58–83–76–1–18–38 an achter Stelle (freundlicher Hinweis Giona ta Brusa). (Ir) nur Signaturen (s. bei G). (Iv) leer. (1ra–88ra) Marsilius ‹de Inghen›: Quaestiones parvorum naturalium Aristotelis (Lohr 331–332, Nr. 15). Tit. (18. Jh., am oberen Blattrand, vom Schreiber des Besitzvermerkes auf derselben Seite): De passionibus Magistri Marsilii. Darüber ältere Angabe: magistri Marsilii. Inc.: [U]trum de passionibus et operacionibus animatorum ponenda sit sciencia distincta a sciencia de anima de animalibus et a sciencia de plantis … – Expl.: … circa hanc materiam de quo sit Deus in secula seculorum benedictus Amen. 276 Cod. 440 Schlussschrift: Expliciunt Questiones parvorum naturalium reverendi magistri Marsilii. (88rb–vb) nur Rahmung. 2 (89ra–268ra) Quaestiones secundum Iohannem Buridanum super libros Ethicorum Aristotelis I–V, unvollständig (Lohr 179–181, Nr. 42). (89ra–91rb) Prolog. Inc. mut. (Platz für Textbeginn freigelassen, nicht ausgeführt): ] Aristoteles 2o Ethicorum cum dicit eam esse non contemplacionis gracia. Iste est prologus quem promittit (!) magister Buridanus libris Ethicorum quo primo innuit primam conclusionem … – Expl.: … malefacere sed tamen non stat eum permanere eum moralem philosophum et eum malefacere. Sequitur questio utrum de virtutibus sit sciencia etc. (91rb–268ra) Text. Inc.: In conclusione prima dicitur quod de virtutibus potest esse sciencia arguitur virtus non est sciencialis … – Expl. mut.: … qui bonus deliquit et bonus qui Aristoteles 2o probatur [ (Lib. V, qu. 14) Incipit und Explicit der Buchanfänge gedruckt in M. Markowski, Buridanica quae in codicibus manu scriptis bibliothecarum Monacensium asservantur. Wroclaw et al. 1981, 119–120, Nr. 15. Parallelüberlieferung: München, UB, 4o Cod. ms. 685, Bl. 1r. (268rb–vb) nur Rahmung. 3 (269ra–346rb) ‹Iohannes Buridanus›: Quaestiones super libros Meteororum Aristotelis, unvoll ständig (Lohr 172, Nr. 21–22). (269ra–318vb) Lib. 1. Inc. mut. (qu. 2): ] deficientem et respondent totum ad temperamentum sanitatis … – Expl.: … ita profunde descendit venit enim mineam de alio loco ad illum et hec sufficiant de illa materia. (319ra–346rb) Lib. 2. Inc.: [I]am de mari composita fuerunt multa super primum librum. Sed adhuc super secundum queritur de perpetuitate vel generacione maris scilicet utrum fuerit per petuum … – Expl.: … violenta et impetuosa. Soluciones aliarum racionum satis haberi possunt ex predeterminacionis etc. Schlussschrift: Et sic est finis libri Meteorum. (346va–b) nur Rahmung. (347ra–411rb) Lib. 3. (347ra) Prolog. Inc.: [R]estat querere aliqua circa 3m librum et verum est quod ab antiquo sole mus illum totalem librum dividere in 4or parciales … – Expl.: … circa 3m librum queremus utrum etc. (347ra) Text. Inc.: Queritur primo utrum et Nephie et Typhones descendunt ad terram ex nu bibus … – Expl.: … de aliis. Et sic est finis questionum primi 2i et 3i librorum Metheororum reverendi magistri Iohannis Buridani etc. (387va–b) nur Rahmung (ohne Textverlust). (411va–423vb) Lib. 4. Inc.: [C]irca quartum Metherorum (!). Utrum tamen 4or sunt qualitates prime scilicet caliditas frigiditas siccitas et humiditas. Et arguitur primo quod non quia … – Expl.: … quia non uritur ab igne dicimus quod tale animal habitat in igne etc. et tamen de isto. Schlussschrift: Et sic est finis. (424r–v) nur Rahmung. A. P. / U. S. Cod. 447 277 Cod. 447 KANONISTISCHE SAMMELHANDSCHRIFT, u. a. ALBERTUS DE BRIXIA Pap. 208, X* Bl. 295 × 215. Süddeutscher Raum, 1414. B: Am seitlichen Blattrand Wasserschaden, vereinzelt kleine Löcher, Blätter der ersten Lage am Vorderschnitt tw. leicht eingerissen. Bl. 1 und 12 lose, Bl. 133 und 134 Riss. Wasserzeichen s. WZMA. Wasserzeichenbe fund mit Datierung übereinstimmend. Lagen: 5.VI60 + V70 + 3.VI106 + V116 + 3.VI152 + V162 + 3.VI198 + V208 + VX. Bl. X klebte urspr. als Spiegel auf dem HD (Rostspuren). Lagenreklamanten, meist stark be- bzw. weggeschnitten, sowie Kustoden in schwarzen oder roten arabischen Ziffern, tw. beschnitten. Am unteren Blattrand Zählung der Blätter der ersten Lagenhälfte, bisweilen beschnitten. In den Lagenmitten Pergament fälze, meist unbeschrieben, in fünfter und neunter Lage beschrieben in karolingischer Minuskel in Rot: quis descendit de celo filius / hominis qui est in celo (Io 3,13) bzw. Quis accendit lucernam (Lc 11,33). Um die letzte Lage Pergamentfalz. Bl. 1r–13v, 20r–v, 41v–43r, 50r, 52v, 71r, 83r, 86v, 127v, 163r, 205r am oberen Blattrand Angabe des jeweiligen Buches in Rot, am seitlichen Blattrand Kapitelzählung mit Buchstaben. Am oberen Blattrand zeitgenössische Foliierung in roten arabischen Ziffern von 1–208, anschließend Foliierung 19. Jh. in römischen Zahlen I–VIIII durch die ULB Tirol. Letztes Blatt (X) ungezählt. Spuren von nicht mehr vorhandenen Signakeln. S: Schriftraum 215/220 × 150. Zwei Spalten, von Tintenlinien gerahmt, zu 38–43 Zeilen. Bastarda, dat. 29. Juli 1414 (Bl. 206vb bzw. Bl. IXra). Schreiber: Hainricus Chirchmair de Malhenstorff (Bl. 206vb) (möglicher weise Kloster Mallersdorf OSB, Diözese Regensburg). Bl. IXra Nachtrag von späterer Hand in Notula. Bl. Xr: Schriftraum 235 × 165, einspaltig (ohne Rücksicht auf die vorhandene Rahmung), zu 58 Zeilen. Bl. 207ra–208vb sowie Xr flüchtigere Bastarda von zeitgenössischer Hand. Ergänzungen am unteren Blatt rand von späterer Hand. A: Rote Auszeichnungsstriche, selten Paragraphzeichen, Durch- und Unterstreichungen, Rahmungen sowie Über- und Schlussschriften. Zweizeilige rote Lombarden, vereinzelt mit Punktverdickungen. Bl. 1ra vier zeilige rot-schwarze Initiale, bogenförmig gespalten, mit flüchtigem, rotem Dekor im Binnenfeld. Bl. 207ra– 208vb sowie Bl. Xr keine Rubrizierung. Bl. 43va und 44ra am unteren Blattrand Repräsentanten, tw. beschnitten. E: Gotischer Einband: rot gefärbtes Schafsleder über dickem Holz mit Blindlinien. Süddeutscher Raum, 1. H. 15. Jh. Leder stark abgerieben, am VD und HD eingerissen. Holz des HD vertikal gebrochen. VD und HD gleich: ein von zweifachen Streicheisenlinien gebildeter Rahmen, Mittelfeld durch zweifache Streicheisenlinien diagonal und in der Mitte waagrecht unterteilt. In der Mitte sowie an den Ecken Spuren von fünf runden Beschlägen. Reste von zwei Langriemen-Leder-Metall-Schließen. Kanten gerade. Rücken: drei Doppel bünde. Im ersten Feld aufgeklebtes Papiertitelschild M: S: 1414 Liber de Officio, im zweiten Signaturschild (s. bei G), im vierten aufgeklebtes Papierschild, leer. Kapitale mit rot gefärbtem Leder umstochen. Am Spiegel des VD Signaturen, Exlibris (s. bei G) sowie Abklatsch eines wohl abgelösten beschriebenen Blattes, Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Bl. 1r Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r alte Signatur der ULB Tirol II4 C 22 (?) (Tinte, radiert), am Spiegel des VD weitere Signaturen der ULB Tirol II`2 C 6 (Bleistift) und 447 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 447. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm II 107. – Stegmüller RS Nr. 368. – Kaeppeli Nr. 98. – Stefani 108. – Peintner, Stiftsbibliothek 121. – Katalog ULB Tirol V 330, 673. 1 (1ra–206vb) ‹Albertus de Brixia›: Summa de officio sacerdotis (Kaeppeli Nr. 98. Bloomfield Nr. 5053, vgl. Nr. 5045). Tit. (am oberen Blattrand von anderer Hand ergänzt): Incipit. De fide, caritate, iusticia, tem perancia. Tytulus libri est summa de officio sacerdotis. Inc.: Quoniam ut ait Ieronimus sacerdotis officium est respondere de lege et ut dominus dicit per Malachiam prophetam Labia sacerdotis custodiunt sacramentum et legem … – Expl.: … faciens nos filios suos adoptivos per unigenitum filium dominum nostrum Ihesum Christum qui cum patre 278 Cod. 447 et spiritu sancto eternaliter est Deus unus et verus ipsi Deo sit honor et gloria qui vivit in eternum et regnat per omnia secula seculorum Amen. Deo gracias etc. (1ra) Lib. 1. – (42va) Lib. 2. – (78ra) Lib. 3. Schlussschrift mit Schreibernennung und Datierung: Finitus est liber iste per manus Hainrici Chirchmair de Malhenstorff dominica Ecce Deus adiuvat Anno domini Mo quadringentesimo quartodecimo etc. (29. Juli 1414). Lit.: Schulte GQ II 424; M. Grabmann, Mittelalterliches Geistesleben. Abhandlungen zur Geschichte der Scholastik und Mystik III. München 1956, 323–351: Albert v. Brescia und sein Werk „De officio sacerdotis“ (tw. Textabdruck); Ders., Albert von Brescia OP (gest. 1314) und sein Werk „De officio sacerdotis“. Ein Beitrag zur Geschichte der seelsorgerlichen Verwertung der Summa theologica des hl. Thomas in der älteren italienischen Dominikaner- und Thomisten schule. Divus Thomas (Friburgensis) 18 (1940) 5–38. 2 (207ra–208ra) De usura. Inc.: Utrum usura mortaliter culpabilis sit penes intencionem principaliter mensuralis (!). Argu itur quod non quia usura attenditur penes pactum mutui inter diversas personas contractum ergo non penes intencionem principaliter … – Expl.: … quod scriptura sacra et cetera sancta affert. Inc. wie Prag, Národní knihovna České republiky, IX B 7, Bl. 372v–373v (Expl. mut.). 3 (208ra–vb) De reemptionibus. Inc.: Casus talis est. Quedam domus religiose quarum fratres non manducant emerunt pensiones annuas perpetuas ipsis oportunas … – Expl.: … prudentes esse conamur. Mit Gutachten von (208ra) Hainricus de Duren advocatus in curia Coloniensi; (208rb) Henricus Stuger de Bonna (!), decretorum doctor; Iohannes Wirborgensis, decretorum doctor; Iohannes de Novo Lapide, legum doctor; Iohannes de Cerno, legum doctor; (208va) Theodericus de Monasterio, sacro (!) theologie professor Colonie; Iacobus de Susato ordinis Predicatorum sacre pagine professor; Nicolaus de Spiria, sacre theologie professor fratrum ordinis Camelitarum (!); Iohannes Ban., legum doctor prepositus ecclesie sancti Georgii Coloniensis; Petrus cardinalis Cameracensis; Iohannes de Gersano, cancellarius Parysiensis; cardinalis Florus; (208vb) archiepiscopus Ianuensis; Angelus rector ecclesie sancti purati Utinensis dyocesis decretorum doctor. Parallelüberlieferung: Frankfurt, Staats- und Universitätsbibl., Fragm. lat. IV 2; Nürnberg, Stadt bibl., Cent. VI 11, Bl. 359r. Darunter: Sequitur confirmacio bulle super hiis dictis in fine libri. 4 (Ira–IXra) Register zu Nr. 1. Mit Blattangabe (rechts) und Kapitelzählung (links). (IXra) Schlussschrift mit Datierung: Et sic est finis etc. anno etc. XIIIIo. 5 (IXra) Nota de baptismo. Inc.: Queritur quis effectus baptismi. Respondetur secundum Thomam in 4o Pueri quidem in baptismo mundantur … – Expl.: … et parentes confirmati. (IXrb–v) leer. 6 (Xr) Martinus V.: Bulla „Regimini universalis“, Rom, 1425 Juli 2 (Extrav. Com. Lib. 3, tit. 5, cap. 1. CIC, ed. Friedberg II 1269–1271). (Xv) nur Rahmung. U. S. Cod. 453 279 Cod. 453 MEDIZINISCHE SAMMELHANDSCHRIFT Pap. III, 234, I* Bl. 295 × 210. T. I: Wiener Raum, um 1455–1460, T. II–IV: Wiener Raum (?), um 1440. B: Vier Teile: I (III, 1–58), II (59–142), III (143–186), IV (187–234, I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. E: Schmuckloser Einband: dunkelbraunes Leder über Holz. S. Maria di Rosano (Firenze), Ende 20. Jh. Einband sowie Bindung im Zuge der Restaurierung erneuert. Kanten gerade. Rücken: drei Doppelbünde. Im ersten Feld Spuren eines aufgeklebten Signaturschildes, im vierten Signaturschild (s. bei G). Kapitale mit hellblauem und rotem Zwirn umstochen. Spiegel Pap. 20. Jh. Am Spiegel des VD Fragment mit Besitz vermerk (s. bei G) sowie Fragment mit Notiz (s. bei Inhalt) eingeklebt. Am Spiegel des HD Etikett der Restaurierwerkstätte (s. bei G). Bl. IIIv Titelschild (möglicherweise einst am urspr. Einband aufgeklebt) Tegni Galieni. Libri afforismorum. G: Wohl Geschenk des Neustifter Präbendars und Klosterarztes Bertoldus de Saxonia ans Stift (gemeinsam mit Cod. 123 und vermutlich auch dem tw. von denselben Händen geschriebenen Cod. 748, vgl. Schenkungsver merk in Cod. 748, Bl. IIIv und Cod. 123, Bl. Vv). Auf dem Titelschild (eingeklebt auf Bl. IIv) unter dem Titel in roter Tinte T9, möglicherweise alte (Neustifter?) Signatur oder Besitzvermerk. Am Spiegel des VD eingeklebtes Fragment mit Besitzvermerk 15. Jh. Iste liber est monasterii sancte Marie virginis ad Novam cellam Brixinensis dyocesis. Bl. 143r Besitzvermerk 15. Jh. Iste liber est monasterii sancte Marie virginis in Novacella. Bl. 1r Besitzvermerk 18. Jh. Collegii Neocell. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro S. Maria di Rosano), kein Restaurierbericht vorhanden. Bl. Ir Signatur 20. Jh. Ms 453 (Bleistift). Am Rücken Signaturschild 20. Jh. 453. L: Kristeller I 439. – P. Kibre, Hippocrates Latinus. Repertorium of Hippocratic writings in the Latin Middle Ages. New York 21985, II 277 und III 269. – Katalog ULB Tirol V 348, 674. I (III, 1–58) B: Pap. tw. fleckig, insbes. Bl. 1r. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasser zeichenbefund. Identische Wasserzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 187 (dat. 1458); Wien, ÖNB, Cod. 3901 (dat. 1461). Lagen: (I–1)I (Vorsatzbl. 20. Jh.) + IIII (Vorsatzbl. 15. Jh.) + 4.VI48 + V58. Ge genbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Bl. 12v Lagenreklamante erhalten. Bl. 39r–53r am oberen rechten Blattrand der Rectos Zählung der Textabschnitte (prima particula Amphorismorum [!], 2a particula etc.). S: Bl. 1r–33r Schriftraum 180 × 110, von Tintenlinien gerahmt, zu 23–25 Zeilen. Bl. 33v–34v Schriftraum 245 × 150, von Tintenlinien gerahmt, zu 70–74 Zeilen. Bl. 35r–53r Schriftraum 200 × 120/125, von Tinten linien gerahmt (Bl. 50v–53r nur seitlich begrenzt). Bl. 35r–38v zu 37–38, Bl. 39r–46v zu 24–27, Bl. 47r–53r zu 23 Zeilen. Bl. 53v–58v Schriftraum 230/240 × 160. Zwei Spalten, von Tintenlinien gerahmt, zu 50–59 (Bl. 53v–54v zu 72) Zeilen. Bastarda von drei Händen: 1) Bl. 1r–46v; 2) Bl. 47r–53r; 3) Bl. 53v–54v und Zählung der Textabschnitte auf Bl. 39r–53r (= späterer Duktus des Schreibers von T. II–IV, von Neustift, Stiftsbibl., Cod. 748, T. I–III und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identi fizieren mit Bertoldus de Saxonia, s. bei G). A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Überschriften. Ein- und zweizeilige (Bl. 35r: dreizeilige) rote Lombarden, meist mit Punktverdickungen. Bl. 1r sowie Bl. 39r–46v Platz für ein zeilige Lombarden freigelassen, nicht ausgeführt. Repräsentanten. Bl. 53va–54va keine Rubrizierung. 1 (VDS, aufgeklebtes Schildchen) Besitzvermerk (s. bei G). Notiz: Prima pronosticorum: ffinis laudabilis medicine existit cum actor futura in singulis propendit quamvis omnes minime curare possit. Quod si posset non iam futuri previsor sed divinis prophetis nobilior diceretur. (Ir–IIIr) leer bis auf Signatur (s. bei G). 280 Cod. 453 (IIIv) Titelschild (s. bei E) sowie Notiz zum Inhalt 18. Jh. Regni (!) Galieni et Joannis Damas ceni Aphorismi. 2 (1r–33r) Galenus: Ars parva seu Microtegni, tr. Gerardi Cremonensis (Druck: GW 2678–2683. – Thorndike–Kibre 1585), mit Interlinear- und Marginalglossen, tw. mit Paraphrase (Thorndike– Kibre 700). Tit.: Incipit Tegni Galieni de corporibus sanis egris et neutris et signis sanis egris et neutris. (1r) Prolog. Inc.: [T]res sunt omnes doctrine que ordine habentur. Prima quidem ex finis no cione … – Expl.: … sed explicacionem demonstratorum dicentur nunc (rot:) de significacione medicine. (1v) Text. Inc.: Medicina est sciencia sanorum egrorum et neutrorum … – Expl.: … scribens et quot sint et qualia determinabo in hiis oracionem. Glosse. Inc.: Nota secundum Averroim quod medicina est de subiecto cuius subiecti racio in medicina salvatur … – Paraphrase. Inc.: In omnibus doctrinis que secundum ordinem currunt incessus sunt secundum rerum ordines. Una earum est que fit secundum … 3 (33v–34v) Divisiones libri Tegni Galieni. Inc.: Tres sunt omnes doctrine que ordine habentur etc. Istius libri qui Tegni id est pars (!) parva dicitur … – Expl.: … in tradicione istius libri visus est observasse. Schlussschrift: Expliciunt divisiones libri Tegni Galieni. 4 (35r–38v) Iohannes ‹Mesue›: Liber aphorismorum (Ed.: D. Jacquart, G. Troupeau, Yûḥannâ ibn Mâsawayh [Jean Mesue], Le livre des axiomes médicaux [Aphorismi]. Genève, Paris 1980, 107–223. – Thorndike–Kibre 824). Tit.: Incipit Liber afforismorum Iohannis Damasceni. 5 (39r–53r) ‹Hippocrates›: Aphorismi (Thorndike–Kibre 1704. Kibre, Hippocrates Latinus II, Nr. III, mit Nennung der Hs.), tr. anonyma (P. Kibre, Hippocrates Latinus II, Nr. III,3). Mit Inter linear- und Marginalglossen Bl. 39r–42r. Text. Inc.: [V]ita brevis ars vero longa tempus acutum experimentum vero fallax iudicium autem difficile … – Expl.: … quanto magis dabuntur tanto plus peyorabuntur. Glosse. Inc.: Continuo oportet in medicina libros afforistica brevitate collaciones componere … 6 (53va–54va) Quaestiones. (53va) Inc.: Utrum in omni tempore morbi possit fieri evacuatio in febre putrida. Quod sic auc toritate illius afforismi. Inchoantibus morbis etc. … – Expl.: … melius et perfectius complebitur cum adiutorio artis. (54va) Inc.: Utrum corpus discrasiatum ex generacione et tempore debeat reduci ad tempera mentum … – Expl.: … que requirit calefaciencia vel subtiliancia vel incidencia. (54vb–58vb) leer. II (59–142) B: Pap. tw. leicht fleckig. Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identi sche Wasserzeichen: Neustift, Stiftsbibl., Cod. 748, T. III. Lagen: 7.VI142. Ab Bl. 71r jeweils am Lagen beginn am oberen Blattrand mittig Kustoden, tw. ausgeschrieben, tw. in arabischen Ziffern, beschnitten und tw. radiert (3–6, 7us). Am oberen Blattrand rechts Zählung der Textabschnitte (beginnend Bl. 60r mit prima particula) und mittig Foliierung innerhalb der einzelnen Abschnitte. S: Schriftraum 240/250 × 155/160. Zwei Spalten, seitlich von Tintenlinien begrenzt, zu 50–59 Zeilen. Bastarda (= Schreiber von T. III und IV, von Neustift, Stiftsbibl., Cod. 748, T. I–III und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Abschnitts anfänge in Textualis formata (Textura). A: Rote Auszeichnungsstriche, Paragraphzeichen und Durchstreichungen. Abschnittsanfänge gerahmt. Cod. 453 7 281 (59ra–134ra) ‹Iacobus Forliviensis›: Expositio in Hippocratis Aphorismos (Druck: GW M10637. – Thorndike–Kibre 1087. Kibre, Hippocrates Latinus III 269). Inc.: [V]ita brevis ars vero longa tempus acutum. Pretermissis quibusdam superfluis inquisicio nibus in principio huius libri … – Expl.: … causam scilicet frigiditatem et humiditatem. Ideo non procedit contra dicta. (134rb) nur Rahmung. (134v–142v) leer. III (143–186) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 534, T. I (dat. 1439); Neustift, Stiftsbibl., Cod. 748, T. III. Lagen: 2.VI166 + 2.V186. Ab Bl. 143r am Lagenbeginn am oberen Blattrand mittig Kustoden (primus–4). Bl. 143r–165r am oberen rechten Blattrand der Rectos Kennzeichnung des ersten Textabschnittes (prima). S: Schriftraum 225/240 × 155/160. Zwei Spalten, seitlich von (tw. zweifachen) Tintenlinien begrenzt, zu 52–58 Zeilen. Zirkelstiche. Bastarda (= Schreiber von T. II und IV, von Neustift, Stiftsbibl., Cod. 748, T. I–III und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Beginn der Quaestiones in Textualis formata (Textura). A: Rote Auszeichnungsstriche, Paragraphzeichen und Durchstreichungen. Überschriften gerahmt. 8 (143ra–183ra) Quaestiones in Hippocratis Aphorismos, unvollständig (Kibre, Hippocrates Lati nus III 269). Text bricht in Particula 6 ab. Tit.: Circa primam afforismorum. In exercitantibus ad summum. – Inc.: Utrum sit possibile hu morem vel humores nostri corporis peccare solum in quali. Et arguitur quod non. Primo sic qui libet humor peccans … – Expl. mut.: … quia non video secundum primam reversionem quomodo responderetur [ (183rb–186vb) nur Rahmung. IV (187–234, I) B: Pap. fleckig, tw. Tintenkleckse. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Was serzeichenbefund. Verwandte Wasserzeichen: Neustift, Stiftsbibl., Cod. 748, T. I. Lagen: (VI+1)199 + 2.VI223 + (VI–1)234 + (I–1)I* (Nachsatzbl. 20. Jh.). Gegenbl. zum Nachsatzbl. als Spiegel auf dem HD aufgeklebt. Bl. 190 kleineres eingehängtes Einzelbl. (230 × 210). Ab Bl. 200 am Lagenbeginn am oberen Blattrand mit tig Kustoden (2–4). S: Schriftraum 230/235 × 155/160. Zwei Spalten, seitlich von Tintenlinien begrenzt, zu 52–55 Zeilen. Zirkel stiche. Bastarda (= Schreiber von T. II und III, von Neustift, Stiftsbibl., Cod. 748, T. I–III und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Bl. 187ra–b und 191ra Beginn der Quaestiones in Textualis formata (Textura). A: Rote Auszeichnungsstriche, Paragraphzeichen und Durchstreichungen. Überschriften gerahmt. 9 (187ra–232rb) ‹Iacobus Forliviensis›: Quaestiones primi tegni Galeni (Druck: GW M10647. – Thorndike–Kibre 1676). Inc.: [U]trum tantum tres sint omnes doctrine ordinarie. Arguitur quod non. Primo quot sunt questiones tot sunt sciencie vel doctrine … – Expl.: … sex rerum non naturalium amministracio sub qua eciam acta anime continentur. (232va–234vb) nur Rahmung. (Ir–v) leer. L. Š. 282 Cod. 483 Cod. 483 HONORIUS AUGUSTODUNENSIS Perg. I, 76, I Bl. 255 × 170. Entstehungsort unbestimmt, 2. H. 12. Jh. B: Perg. von unterschiedlicher Qualität, leicht fleckig, abgegriffen, vereinzelt Löcher und Nahtspuren. Lagen (Neubindung 20. Jh.): (I–1)I (Vorsatzbl., Pap. 20. Jh.) + II4 + IV12 + III18 + 2.I22 + IV30 + II34 + 2.I38 + 3.IV62 + I64 + IV72 + II76 + (I–1)I* (Nachsatzbl., Pap. 20. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Urspr. wohl durchwegs Quaternionen, zahlreiche Fehlstellen (s. bei Inhalt): zw. Bl. 2 und 3 (Lage 1) fehlen innere Doppelbl., zw. Bl. 12 und 13 sowie Bl. 18 und 19 (Lage 3) äußerstes Doppelbl., zw. Bl. 20 und 21 (Lage 4) äußere Doppelbl., zw. Bl. 21 und 22 (Lage 5) innere Doppelbl., zw. Bl. 32 und 33 (Lage 7) innere Doppelbl., zw. Bl. 34 und 35 sowie 35 und 36 (Lage 8) innere und äußere Doppelbl., zw. Bl. 36 und 37 sowie 38 und 39 (Lage 9) äußere Doppelbl., zw. Bl. 63 und 64 (Lage 13) i nnere Doppelbl. Fehlbindung: urspr. Lagen I–VI (Bl. 13–36) heute nach urspr. Lage VII (Bl. 5–12) eingebunden, vgl. Kustode Bl. 12v VIIus, Lage 3 (Bl. 13–18) Blattzählung am rechten Rand von zeitgenössischer Hand 2–7 sowie Inhalt. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 205/215 × 130/145, von seitlich zweifachen Blind- bzw. Bleistiftlinien gerahmt. Zirkelstiche. Bl. 1r–38v zu einheitlich 33 Zeilen, ab Bl. 39r zu einheitlich 30 Zeilen auf Blind- bzw. Bleistiftlinierung. Karolingische Minuskel mit unterschiedlichem Duktus von zwei Händen: 1) Bl. 1r–38r; 2) Bl. 38v–76v. Bisweilen Tinte radiert und verblasst. Vereinzelt Randnotizen, tw. von späterer Hand. A: Rote Paragraphzeichen (vereinzelt) und Überschriften. Bl. 19v und 22r Zeigehände. Ein- und zweizeilige rote Lombarden, vereinzelt mit Punktverdickungen und Ausläufern. Ab Bl. 39r Repräsentanten am seit lichen Blattrand, quer zum Schriftraum. E: Bibliothekseinband: dunkelrot gefärbtes Leder über Holz. Rignano sull’Arno (Prov. Firenze), um 1980. Einband und Bindung im Zuge der Restaurierung erneuert. Kanten gerade. Rücken: fünf Doppelbünde. Im ersten Feld Spuren eines Signaturschildes, im fünften weißes Signaturschild (s. bei G). Kapitale mit natur farbenem und gelbem Garn umstochen. Spiegel Pap. 20. Jh., leer bis auf Etikett der Restaurierwerkstätte am HD (s. bei G). G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Auf hebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des HD Nachweis der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro S. Maria di Rosano), kein Restaurierbericht vorhanden. Bl. Ir Signatur 20. Jh. Ms. 483. Am Rücken Signaturschild 20. Jh. 483. L: R. Foreville, J. Leclercq, Un débat sur le sacerdoce des moines au XIIe siècle, in: Analecta monastica IV (Studia Anselmiana 41). Roma 1957, 8–118. – Katalog ULB Tirol V 542, 674. (Ir–v) leer bis auf Signatur (s. bei G). (1r–76v) ‹Honorius Augustodunensis›: Gemma animae, unvollständig (PL 172, 541–753). Anordnung in Hs. vielfach vom Druck abweichend (möglicherweise aufgrund Neubindung), Kapitel einteilung und Überschriften tw. abweichend, Kapitel vereinzelt leicht gekürzt, bisweilen zusätzliche Einschübe. (1r–2v) Lib. 2, cap. 1–15 (PL 172, 615D–620D). Inc. mut.: ] exercitus in militiam congregatur. Signa militibus per turbas (!) dantur … – Expl. mut.: … in bonis in locum exul[tationis adducet (3r–8v) Lib. 2, cap. 50–68 (PL 172, 629C–642C). Inc. mut.: ] etiam Benedictus in eadem significatione … (8v–12v) Lib. 3, cap. 1–30 (PL 172, 641D–650C). Expl. mut.: … Unde in illo tempore alleluia cantatur [ Mit Auslassung in cap. 22 (De tribus nuptiis …), s. Bl. 67r–v. (13r–18v) Lib. 1, cap. 3–38 (PL 172, 544A–555D). Inc. mut.: ] subiugat. Oppressos a diabolo liber ans … – Expl. mut.: … duo typum Enoch et Helye preferunt qui Iudaicum [ (19r–20v) Lib. 1, cap. 63–73 (PL 172, 563C–567A). Inc. mut.: ] sumit significans angelum qui Christo resurgente lapidem … – Expl. mut.: … ad monasterium redimus quasi ad locum certa[minis (21r–v) Lib. 1, cap. 46–51 (PL 172, 557D–559D). Inc. mut.: vir]gine nascitur et per multas passiones in candorem resurrectionis … – Expl. mut.: … scriptis vocibus premonstraverunt [ Cod. 483 283 (22r–32v) Lib 1, cap. 84–149 (PL 172, 570C–590C). Inc. mut.: ] Missa legatio dicitur Totum tempus quo Christus mundum intravit … – Expl. mut.: … diversi libri sacre scripture. Secretum quod claustri gerit [ Mit Einschub zw. cap. 114 und 115 (Bl. 28r): Inc.: Licet laico plus in die … – Expl.: … aliis sacerdotibus licet. (33r–34v) Lib. 1, cap. 174–187 (PL 172, 597D–601D). Inc. mut.: ] Dei dicuntur quia ex precepto Dei in servitium Dei offeruntur … – Expl. mut.: … cum Christiani relicta lege Dei paganos spurcis [ (35r–v) Lib. 1, cap. 58–63 (PL 172, 561D–563C). Inc. mut.: ] ecclesia conficitur et capiti Christo con iungitur … – Expl. mut.: … diaconus calicem tenet dum papa sanguinem [ (36r–v) Lib. 1, cap. 73–79 (PL 172, 567A–568D). Inc. mut.: certa]minis tendimus. Scrinium cum reli quiis portamus contra demones … (37r–38r) Lib. 4, cap. 2–13 (PL 172, 690C–693C). Inc. mut.: ] rubrum id est per baptismum vel per desertum id est penitentiam … (38r–v) Lib. 3, cap. 33–38 (PL 172, 651C–652B). Expl. mut.: Nabuchodonosor est diabolus. [ Mit Ein schub nach cap. 37: Inc.: Nabuchodonosor quippe rex Babilonis bellica manu Ierusalem d estruxit … – Expl.: … duplex gaudium habuerunt. (39r–v) Lib. 3, cap. 46–50 (PL 172, 657A–658D). Inc. mut.: ] adhuc super corda Iudeorum positum et ideo non valent videre splendidum legis sensum … (39v–40r) Lib. 4, cap. 14 (PL 172, 693C–694B). (40r–55r) Lib. 3, cap. 51–149 (PL 172, 658D‑–684D). Mit zwei Einschüben: 1) (53v) zw. cap. 145 und 146: Inc.: Responsorium de ascensione nostri domini … – Expl.: … ad eum super sydera. 2) (54r–v) in cap. 146: Inc.: Ut in aliis vero dominicis. Propter baptismalia sacramenta in tribus personis … – Expl.: … remissio peccatorum percipitur. Quod in hac nocte vel in omnibus summis festivitatibus ad vesperam … (55r–60v) Lib. 4, cap. 15–40 (PL 172, 694B–703C). (60v) Lib. 3, cap. 150 (PL 172, 684D–685A). (60v–63v) Lib. 4, cap. 41–53 (PL 172, 703D–708C). Expl. mut.: … David pro populo alleluia vero in te [ (64r–67r) Lib. 4, cap. 74–89 (PL 172, 718D–724D). Inc. mut.: ] tabernaculorum celebratio ordinati enim … (67r–v) Lib. 3, cap. 22 (PL 172, 648D–649A). Inc. (verkürzt, s. Auslassung Bl. 12r): De tribus nuptiis legitur in evangelio … (67v–76v) Lib. 4, cap. 90–118 (PL 172, 724D–738B). Mit drei Einschüben: 1) (72r–v) in cap. 115: Inc.: Omnibus martyribus prefertur. Sic etiam de loco vel eventu Felicis … – Expl.: … eos coronatos nominabant. Sic de ceteris notandum. 2) (72v–73v) in cap. 116: Inc.: Imperatoris susceptio est tri umphi imitatio. Tryumphus namque regis exhibebatur … – Expl.: … prima propter ornatum secunda est quasi literatura laicorum tercia quia sunt admonitiones clericorum. 3) (76r) kurz vor Ende von cap. 118: Inc.: Mos paganorum versus est in religionem Christianorum … – Expl.: … quia cottidie sollempnia missarum celebrantur. (76v, direkt anschließend, ohne Hervorhebung) Versus de ornamentis filiarum Ierusalem. Inc.: Ecce filie Ierusalem quas delicias vobis preparavi … – Expl.: … ipse vestris nuptiis interesse mereatur. Amen. Parallelüberlieferung: u. a. Düsseldorf, ULB, Ms. B 73, Bl. 94vb, Ms. B 167, Bl. 189va; Berlin, Staats bibl., Preußischer Kulturbesitz, Ms. theol. lat. fol. 348, Bl. 133r; Graz, UB, Hs. 804, Bl. 76v. (Ir–v) leer. A. P. / U. S. 284 Cod. 485 Cod. 485 HUGO DE SANCTO VICTORE Perg. I, 66, I Bl. 240 × 180. Entstehungsort unbestimmt, um 1200. B: Dickes Perg., vereinzelt Löcher und Nahtspuren, Bl. 65v und 66r Wasserflecken. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + (IV–I)6 + 7.IV62 + II66 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsa tzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Lage 9 (= heute Lage 1) unvollständig: urspr. wohl Qua ternio, äußerstes Doppelbl. fehlt (vor Bl. 1 und nach Bl. 6 Textverlust). Am Lagenende Kustoden in römis chen Zahlen, zumeist beschnitten. Lage 2 (Bl. 14v) Kustode Xus, Lage 7 (Bl. 54v) Kustode XVus: Am Beginn fehlen daher wohl acht Lagen. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 185/195 × 125, von zweifachen Blindlinien gerahmt, zu 26 Zeilen auf Blindlinierung. Zirkel stiche. Karolingisch-gotische Mischschrift. Bl. 33v–35r am Blattrand Notizen von späterer Hand. A: Rote Überschriften sowie Kapitelzählung am Blattrand. Meist zwei-, vereinzelt auch ein- bis siebenzeilige rote Initialmajuskeln. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. etwas abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, vier einfache Bünde, von Goldbordüren gesäumt. Im ersten Feld Golddruck M: S:, im zweiten Golddruck DE LAPSU HOMINIS ET SACRAM in Goldrahmung, im dritten Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Bl. 66r Besitzvermerk 15. Jh. Iste liber est sancte Marie in Novacella. Bl. 66v Besitzvermerk 15. Jh. Iste liber est monasterii s. Marie ad Novacellam. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spie gel des VD alte Signaturen der ULB Tirol II4 D 25 (?, Tinte, radiert),2 D 14 S. 2. (Bleistift) und 485 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 485. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol V 547, 674. (Ir–v) leer. (1r–66v) ‹Hugo de Sancto Victore›: De sacramentis, Lib. 1, unvollständig (PL 176, 271D–364A. Ed. R. Berndt, Hugonis de Sancto Victore. De sacramentis Christianae fidei [Corpus Victorinum. Textus historici 1]. Monasterii Westfalorum 2008, 147–269). (1r–11r) Pars 6 (Kapitelzählung in Hs. vom Druck abweichend), unvollständig. (1r–6v) Inc. mut. (cap. 14): revela]cionem plurimum addendum fuit. Modum vero divine cognicionis quam primus ille homo habuisse creditur explicare difficile est excepto eo quod diximus … – Expl. mut. (cap. 26): … in rebus occultis ea que magis probabi[lia (7r) Inc. mut. (cap. 29): ] quam commodum obedientis adtenderet. Itaque ut homo super hanc vitam … (11r) Pars 7. – (24r) Pars 8. – (32v) Pars 9. – (40r) Pars 10. – (51v) Pars 11. – (54v) Pars 12. (Ir–v) leer. U. S. Cod. 495 285 Cod. 495 GREGORIUS MAGNUS Perg. 202 Bl. 210 × 140/145. Entstehungsort unbestimmt, 10. Jh. B: Perg. von unterschiedlicher Qualität, fleckig (Feuchtigkeitsschäden) und abgegriffen, löchrig, Blattränder tw. eingeschnitten. Lagen: 8.IV64 + (III+2)72 + (II+4)80 + (III+2)88 + 2.IV104 + (III+2)112 + IV120 + V130 + IV138 + (III+2)146 + IV154 + (V–2)162 + 5.IV202. Bl. 68, 69, 74, 75, 78, 79, 84, 85, 108, 109, 142 und 143 eingefügte Einzelbl., Gegenbl. zu Bl. 156 und 157 herausgeschnitten (ohne Textverlust). Am oberen Blattrand Foliie rung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: Bl. 155 fälschlich als 195 angeführt. S: Schriftraum 160 × 100/110, seitlich von zweifachen Blindlinien begrenzt, zu 20, Bl. 133–136, 140–157 und 162 zu 19 Zeilen auf Blindlinierung. Zirkelstiche. Karolingische Minuskel in unterschiedlicher Federdicke, wohl von mehreren einander abwechselnden Händen. Bl. 18r, 39r am unteren Blattrand Ergänzungen von der Hand des Schreibers. Bl. 1r Überschrift und Initium in Capitalis rustica. A: Rote Kapitelzählungen, Über- und Schlussschriften, Nennung der Dialogpersonen (tw. mit Repräsentanten). Satzmajuskeln in Lombardenform sowie einzeilige, am Beginn der Bücher dreizeilige rote Initialmajuskeln. E: Schmuckloser romanischer Einband: stark gedunkeltes, fleckiges Schweinsleder über Holz. Entstehungsort unbestimmt, 11. Jh. Am VD schwarze Titelaufschrift in Textualis formata (Textura) 12. Jh. Dyalogus Gregorii de Sunaburg. Spuren zweier Schließen, am VD Dorne erhalten. Kanten leicht abgeschrägt und gerade. Rücken: blau gefärbt (wie öfters in Neustift), zwei Fitzbünde und zwei Doppelbünde. Im ersten Feld weiße Aufschrift Gregorii Dialogi MS., im zweiten Signaturschild (s. bei G), im vierten aufgeklebtes Papierschild, leer. Kapitale mit Spagat umstochen. Spiegel des VD und HD: gedunkelte, beschriebene, jedoch nicht mehr lesbare Pergamentstreifen, tw. vom eingeschlagenen Rückenleder überdeckt, am Spiegel des VD Exlibris und Signaturen (s. bei G). G: Aufgrund der Titelaufschrift am VD ab dem 12. Jh. im Besitz des Benediktinerinnenstifts Sonnenburg im Pustertal. Zeitpunkt und Art der Erwerbung durch Neustift unklar. Am Spiegel des VD Exlibris der Stifts bibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II 1 B 17 (Tinte, radiert) und II2 E 10 (Bleistift). Am Rücken gelbes Signaturschild der ULB Tirol 495. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol V 610, 674. Gregorius Magnus: Dialogi = Dialogorum libri IV de vita et miraculis patrum Italicorum, unvollständig (PL 77, 149–429 [Lib. 1, 3 und 4] bzw. PL 66, 126–204 [Lib. 2]. Ed. U. Moricca [Fonti per la storia d’Italia 57]. Roma 1924. Ed. A. de Vogüé [Sources chrétiennes 260]. Paris 1979 [Buch 1–3] und [Sour ces chrétiennes 265]. Paris 1980 [Buch 4]. – CPL Nr. 1713). (1r) Tit.: Dialogorum Gregorii Papae urbis Romae libri Numero quatuor de miraculis Patrum Itali corum. (1r) Prolog. – (4r) Kapitelübersicht zu Lib 1. – (4r) Lib. 1. – (43v) Kapitelübersicht zu Lib. 2. – (45r) Lib. 2. Bl. 76v/77r (Lagenmitte) Unterbrechung ohne Textverlust (PL 66, 184B6–8): Seitenbeginn Bl. 76v: ad monasterium omnipotentis Dei famulum repperit. Quia (Rest der Zeile leer), auf neuer Zeile: a creditore suo (Rest der Seite leer), Forts. Bl. 77r: pro duodecim solidis graviter affligeretur intimavit … (88v) Lib. 3. – (147r) Lib. 4. Expl. mut. (stark zerstört, Schrift endet am Seitenende): … ad exitum veniens non pertimescat Quando etsi iam novit quod egit [ (PL 77, 405C3 in cap. 45) W. N. 286 Cod. 532 Cod. 532 SAMMELHANDSCHRIFT Perg. I, 145, I Bl. 150 × 110. Entstehungsort unbestimmt, 14. Jh. B: Perg. tw. genäht, Fehlstellen im Zuge der Restaurierung 20. Jh. ergänzt (s. bei G). Lagen: (I–1)I (Vorsatzbl., Pap. 20. Jh.) + 15.IV120 + II124 + 2.IV140 + (III–1)145 + (I–1)I* (Nachsatzbl., Pap. 20. Jh.). Gegenbl. zum Vorbzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 145 herausgeschnitten (Text verlust, vgl. Reklamante). Am Ende der Lagen 1–4 sowie 17–18 schwarz-rot gerahmte Reklamanten mit einfachem Dekor, am Ende der Lagen 5–16 Kustoden in roten römischen Zahlen, bisweilen beschnitten. Zeitgenössische Foliierung in arabischen Ziffern 4–149. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Bl. 1r–122v und 125r–144v: Schriftraum 115 × 75, von Tintenlinien gerahmt, zu 34 Zeilen auf Tintenlinie rung. Zirkelstiche. Textualis von zwei Händen: 1) Bl. 1r–122v und 125r–126r in abwechselnder Größe; 2) Bl. 126v–144r. Nachtrag Bl. 123r–124v: Schriftraum 120 × 75, von Tintenlinien gerahmt, zu 25 bzw. 27 Zeilen. Zirkel stiche. Bastarda 15. Jh. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Rahmungen, Auctoritatesangaben und Überschriften. Bl. 79v Randmarkierung in der Art eines Fleuronnée-Ausläufers. Zeigehände. Lagenrekla manten mit Zierrahmen (Bl. 132v mit Blüte, 140v). Am oberen Blattrand Bezeichnung der einzelnen Bücher in Rot. Ein- bis vierzeilige rote Lombarden. E: Bibliothekseinband: helles Leder über Holz. Rignano sull’Arno (Prov. Firenze), 20. Jh. Einband unter Verwendung von Resten des urspr. Ledereinbandes (15. Jh.) am Rücken sowie Bindung im Zuge der Restaurierung erneuert. Am HD Signaturschild (s. bei G). Kanten gerade. Rücken: drei einfache Bünde, im ersten Feld Signatur schild (s. bei G). Kapitale mit natur- und ockerfarbenem Zwirn umstochen. Spiegel Pap. 20. Jh. Spiegel des VD leer, am Spiegel des HD Etikett der Restaurierwerkstätte (s. bei G). G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Auf hebung des Klosters Neustift der ULB Tirol übergeben. Am Rücken gelbes Signaturschild der ULB Tirol 532. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro S. Maria di Rosano), kein Restaurierprotokoll vorhanden. Am HD Signaturschild 20. Jh. 532. Bl. Ir Signatur 20. Jh. Ms 532 (Bleistift). L: Kaeppeli Nr. 2346. – Bloomfield Nr. 5755. – Katalog ULB Tirol VI 102, 381. 1 (Ir–v) leer bis auf Signatur (s. bei G). (1r–27r) Tractatus de absentis praesentia. (1r) Tit.: Incipit tractatus de gradibus virtutum. (1r) Prolog. Inc.: Cum quadam vice animum meum infelicem vanis ac transitoriis occupacionibus assidue deditum ad meam recogitandam miseriam … – Expl.: … calor vincit teporem longum. (1r) Text. Inc.: Hec ita cogitans menti occurrit illa vocatio trina apostolorum qui primo ad noti tiam … – Expl.: … quidem docentem hominem scientiam. Dominum autem pro laboribus remu nerantem invenire in cursus nostri termino meramur. Amen. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 390, Bl. 91r; München, BSB, Clm 5534, Bl. 73; Augsburg, Staats- und Stadtbibl., 2o Cod. 139, Bl. 258ra. 2 (27r–28v) ‹Ps.-Augustinus›: De praedestinatione Dei (PL 45, 1677–1678. – CPL Nr. 383. CPPM II 180). 3 (28v–34v) ‹Eadmerus Cantuariensis›: De beatitudine coelestis patriae (PL 159, 587B–643B). 4 (34v–122v) ‹Ps.-Henricus de Frimaria› = ‹Ps.-Hugo de Sancto Caro›: In libros I–IV Sententia rum sive Filia magistri (Stegmüller RS Nr. 373. Glorieux Nr. 2bh. Zumkeller Nr. 328d). Cod. 550 287 Inc.: Quoniam velut IIIIor paradisi flumina … Scriptura sacra de duobus agit … Veteris ac nove legis continenciam considerantibus … – Expl.: … per gratiam evaserunt. Schlussschrift mit Schreibervermerk: Finito libro sit laus et gloria Christo. Pitet got umb den der daz geschriben hat. 5 (123r–124r) ‹Richardus de Mediavilla›: Commentarius in Petri Lombardi Sententiarum librum secundum, Prologus (Druck: u. a. GW M22506. – Stegmüller RS Nr. 722, II. Glorieux Nr. 324c). Tit.: […, beschnitten] super secundum librum Sentenciarum etc. – Inc.: Omnia per ipsum facta sunt et sine ipso factum est nihil. Hec verba scripta Io. primo (3) sic possunt exponi. Omnia sci licet entia a Deo alia. Facta sunt per ipsum id est per verbum Dei. Unde Augustinus sermonem dirrigens ad Deum … – Expl.: … coniungantur cum eo. Quod nobis prestare dignetur … per omnia secula seculorum. Amen. 6 (124v) Notiz. Inc.: Nota quod omnis disposicio … – Expl.: … quibus hoc forte meruerunt penitere et hiis similia. 7 (125r–144v) ‹Iohannes Friburgensis›: Confessionale (Druck: GW M13586. – Bloomfield Nr. 5755, mit Nennung der Hs. Kaeppeli Nr. 2346, mit Nennung der Hs.). Tit.: Incipit tractatus de instruccione confessorum. (125r) Prolog. Inc.: Simpliciores et minus expertos confessores de modo audiendi … Anschließend Inhaltsübersicht. (125v) Text. Inc.: Cum aliquid petit suam confessionem audiri quem antea … – Expl.: … et testa mentum (!) celeriter expedient frequentibus admonicionibus impulsabis. Schlussschrift: Explicit liber confessorum. (145r–v) leer. (Ir–v) leer. C. S. Cod. 550 GUIDO DE MONTE ROCHERII. GUILELMUS DURANTIS Pap. I, 446, III Bl. 295 × 210. T. I: Wiener Raum, 6. Jz. 15. Jh., T. II: Raum Brixen, u. a. Villanders, 7. Jz. 15. Jh. (u. a. 1465). B: Zwei Teile: I (I, 1–92), II (93–447[446], III). In der Mitte des oberen Blattrandes zeitgenössische Foliierung 1–87, anschließend Forts. im 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 269 ausgelassen. E: Schmuckloser Einband: dunkelrot gefärbtes Leder über Holz. Rignano sull’Arno (Prov. Firenze), um 1980 (?). Einband unter Verwendung der wohl urspr. Metallteile sowie Bindung im Zuge der Restaurierung erneuert. Zwei Leder-Metall-Schließen: Leder erneuert, Lager und Rasten verziert. Kanten gerade. Rücken: drei Doppelbünde. Im ersten Feld Spuren eines entfernten Signaturschildes, im vierten weißes Signaturschild (s. bei G). Kapitale mit rotem und hellgelbem Garn umstochen. Spiegel Pap. 20. Jh., leer bis auf Etikett der Restaurierwerkstätte am Spiegel des HD (s. bei G). Originaldeckel gelöst und getrennt aufbewahrt (freundlicher Hinweis Simon Terzer, Neustift): rot gefärbtes Leder mit Blindlinien über Holz. Brixen, 15. Jh. (?). Leder abgegriffen, tw. nachgedunkelt und zerkratzt. VD durch zweifache Blindlinien gerahmt, Mittelfeld durch ebensolche Linien diagonal unterteilt. Reste von zwei am oberen Rücken und auf den VD überlappenden Papier(titel?)schildern. Spuren von fünf Beschlägen und zwei Schließen. HD heute Holz großteils freiliegend. Auf den Innenseiten der beiden Holzdeckel jeweils Abklatsch einer Urkunde, tw. unleserlich. Innenseite VD: Notariatsinstrument Brixen 1446, Juni 14 (Io hannes de Radmansdorff clericus Aquilegensis diocesis notarius publicus). Oben alte Signaturen der ULB Tirol: II […] (Tinte, laut Alter Titelkopie an der ULB Tirol: II3 A 9) und II3 D 8 (Bleistift). Innenseite HD: Notariatsinstrument Brixen (?) 1420, Juni 4 (Wolfangus Payr de Hederstorf clericus Pataviensis diocesis publicus imperiali auctoritate notarius). 288 Cod. 550 G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Bl. 1r Besitzvermerk 18. Jh. Collegii Neocell. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Labora torio di restauro del libro, S. Maria di Rosano), kein Restaurierbericht vorhanden. Bl. Ir Signatur (Bleistift) Ms. 550, am Rücken Signaturschild 20. Jh. 550. L: Peintner, Stiftsbibliothek 121. – Katalog ULB Tirol VI 172, 381. – Stefani 110–111. I (I, 1–92) B: Pap. am Beginn sowie am Schluss etwas verschmutzt. Nachweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: u. a. Klosterneuburg, Stiftsarchiv, Rb 25/1 (dat. 1457); Wien, ÖNB, Cod. 4204 (dat. 1458). Lagen: (I–1)I (Vorsatzbl., Pap. 20. Jh.) + 6.VI72 + 2.V92. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Lagenreklamanten, tw. rubriziert. S: Schriftraum 195/205 × 150 (Bl. 1r–24v) bzw. × 140 (ab Bl. 25r). Zwei Spalten, vereinzelt von Blindlinien gerahmt, zu 39–42 Zeilen. Bastarda. Am seitlichen Blattrand Hervorhebung und Wiederholung einzelner Wörter und kurzer Textpassagen sowie Korrekturen von späteren Händen, Bl. 4r–10r in Humanistischer Minuskel in Rot. A: Rote Auszeichnungsstriche und -punkte, Paragraphzeichen, Unter- und Durchstreichungen sowie Bl. 3ra–16ra Überschriften, tw. in schwarzer Tinte ergänzt. Bl. 87vb Schlussschrift in Blau. Zwei- bis vierzeilige, meist dreizeilige rote Lombarden, tw. mit Punktverdickungen, Aussparungen und einfach verzierten Ausläufern, Bl. 86va und 87va Binnenfelder mit einfachem Dekor in Blau gefüllt. Vereinzelt Buchstaben nach Initiale cadellenartig gedehnt und in Rot hervorgehoben (z. B. Bl. 25ra, 47va). Bl. 1ra und 25ra fünf- bzw. sechszeilige rote Lombarden mit fein gezeichnetem tintenbraunem Dekor (als Besatz kammartig abstehende, spitz zulau fende Fadengruppen; diese auch an Cadelle Bl. 25ra). Repräsentanten am unteren Blattrand, tw. beschnitten. 1 (Ir–v) leer bis auf Signatur (s. bei G). (1r) Notiz zu T. II (18. Jh): Rationale Divinerum (!) Officiorum Guilelmi Episc. Minae. 1461. (1ra–88vb) Guido de Monte Rocherii: Manipulus curatorum (Druck: u. a. GW 11716. – Bloom field Nr. 5019. Stegmüller RS Nr. 277). Text bisweilen vom Druck leicht abweichend, vereinzelt Kürzungen bzw. Auslassungen, Erwei terungen bzw. Einschübe. Gliederung der Pars 2 vom Druck abweichend (hier Gliederung vom Druck übernommen). Überschriften zu den einzelnen Abschnitten nur vereinzelt vorhanden, tw. falsch zugeordnet. Überschriften im Register genannt, Reihenfolge jedoch nicht mit Gliederung der Traktate und Kapitel übereinstimmend, vom Druck bes. ab Bl. 38 häufig abweichend. Tract. 3, cap. 6 und 7, sowie Tract. 4, Prolog und cap. 1 fehlen, obwohl laut Register vorgesehen. (1ra) Prolog. Inc.: Reverendo in Christo patri ac domino domino Raymundo divina providencia sancte sedis Valencie episcopo servorum devotorum minimus Gwido de Monte Rocheri cum devota … Quoniam secundum quod dicit propheta Malachias ymo dominus per Malachiam Labia … – Expl.: … ut videant ea que circa suum officium sunt agenda. (2ra–44va) Pars 1. Inc.: Dividitur autem presens opusculum in tres partes in quarum prima agi tur de sacramentis … – Expl.: … ecclesia in suo sensu habundat secundum quod habet in usu sequitur etc. (2ra) Tract. 1. – (3vb) Tract. 2. Mit Einschub am Ende von cap. 2 (4vb–5ra): Inc.: Item si alicui presbytero presentaretur femella et ipse putaret masculum … – Expl.: … baptizatus baptismo flaminis. – (10ra) Tract. 3. – (11vb) Tract. 4. Mit Einschub am Ende von cap. 9 (24vb): Inc.: Notandum tamen etc. quod quidam sacerdos … – Expl.: … datur aliquantulum et pane et vino etc. – (31va) Tract. 5. – (34rb) Tract. 6. – (35rb) Tract. 7,1. – (37rb) Tract. 7,2. (44vb–77ra) Pars 2. Inc.: Verus ac pius samaritanus dulcis Ihesus qui vinum et oleum fundit … – Expl.: … animo sed benigne corrigat et emendet. (44vb) Prolog. – (46va) Tract. 1. – (51va) Tract. 2. – (69rb) Tract. 3. Cod. 550 289 (77ra–87vb) Pars 3. Inc.: Omnium magister et dominus Ihesus Christus de mundo ascensurus ad patrem … – Expl.: … faciat nos rex regum et dominus dominancium Ihesus Cristus. Amen. (87vb) Schlussschrift (blau): Laus Deo Amen. (88ra) De fine huius operis. Ohne Tit. Inc.: Hec circa officium curatorum breviter a me scripta sunt … – Expl.: … et pro me peccatore preces ad dominum fundat. (88ra–vb) Inhaltsverzeichnis, tw. mit Blattangabe (bis Bl. 12). – Schlussschrift: Finitur tabula huius libelli. (89r–92v) leer. Lit.: Schulte GQ II 430. II (93–447[446], III) B: Bl. 96–97 und ab Bl. 352–445 Tintenfraß, tw. Textverlust. Bl. 94–97 und 352–II* im 20. Jh. restauriert (s. bei G). Nachweise und Bilder der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identi sche Wasserzeichen: Neustift, Stiftsbibl., Cod. 155; Innsbruck, ULB Tirol, Cod. 566, T. I (dat. 1463). Lagen: (I+II)98 + 29.VI447(446) + III* (Nachsatzbl., Pap. 15. Jh.) + (I–1)III* (Nachsatzbl., Pap. 20. Jh.). Bl. 94–97 ein gehängter Binio. Gegenbl. zu Bl. III* als Spiegel auf dem HD aufgeklebt. Am oberen Blattrand ab Bl. 99r zeitgenössische Foliierung in Rot, zunächst in arabischen Ziffern (1–9), anschließend in römischen Zahlen (X–LXXXX). Bl. 99, 150, 182, 195, 253, 262, 354 jeweils Spuren eines Signakels. S: Schriftraum 210/230 × 145/150. Zwei Spalten, vereinzelt von feinen Tintenlinien gerahmt, zu 46–48 Zeilen. Schriftraum Register 210/215 × 140. Zwei Spalten, zu 42 Zeilen: Bastarda von drei Händen: 1) Bl. 96ra– 97vb, 180vb–382ra und 385ra–445ra, lokalisiert und dat. Villanders, 1465 (Bl. 97vb und 445ra [24.9.]). Schreiber: Laurencius Smyt (Bl. 445ra); 2) Bl. 99ra–180vb; 3) Bl. 382ra–384vb. Am Blattrand Ergänzungen und Korrekturen tw. von jeweiliger Schreiberhand, tw. von anderer Hand, bisweilen unterstrichen oder ge rahmt. Bl. 180r am Blattrand neuzeitliche Bleistiftnotizen, radiert. A: Rote Auszeichnungsstriche und -punkte, Paragraphzeichen, Durch- und Unterstreichungen, Rahmungen, Überschriften sowie Angaben des jeweiligen Buches am oberen Blattrand, tw. beschnitten und Bl. 201ra–va Hervorhebung der einzelnen Kapitel. Verweiszeichen. Ab Bl. 184r in der letzten Zeile Buchstaben bisweilen gedehnt und Ausläufer in Schleifen gelegt. Zwei- bis sechszeilige, meist vierzeilige (Bl. 262ra achtzeilig, nur durch Umrisslinien) rote Lombarden, meist mit Punktverdickungen, tw. mit Aussparungen, tw. mit Aus läufern (ab Bl. 256vb mehrfach mit Dreiblättern als Besatz), vereinzelt mit Dekor im Binnenfeld: Bl. 117rb Blatt, Bl. 123rb mit ausgespartem Knospenfleuronnée, Bl. 183rb mit rotem Fleuronnée, Bl. 279vb mit ein geschriebenem Gesicht, Bl. 304rb die geometrischen Aussparungen tw. schwarz ausgemalt, Bl. 384vb mit Punktdekor. Bl. 99ra neunzeilige rote Initiale, Binnenfeld in Kompartimente unterteilt und mit Knospengarben gefüllt; Initialfeld, Perlenbesatz und Fadenausläufer (in dreiblättriger Blüte endend) sowie Profilkopf mit Zipfelmütze auf dem oberen Seitenrand recht flüchtig gezeichnet. Repräsentanten. 2 (93r–95v) leer. (96ra–445ra) ‹Guilelmus Durantis›: Rationale divinorum officiorum (CCCM 140–140B). (96ra–97vb) Register. (97vb) Schreibervermerk: Iste liber dicitur Racionale Wilhelmi Minacensis episcopi pro eo quod utitur racione et quia eorum qui fiunt in ecclesiasticis officiis continent (!) raciones. Anno domini (gestrichen: mille) Mo CCCCo sexagesimo quinto finitus est presens liber in Willanders. (98r–v) leer. (99ra–445ra) Tit.: Incipit Racionale divinorum officiorum. (99ra) Prolog. – (100va) Lib. 1. – (124va) Lib. 2. – (135va) Lib. 3. – (150rb) Lib. 4. – (232rb) Lib. 5. – (262ra) Lib. 6. – (396ra) Lib. 7. – (431va) Lib. 8. (445ra) Schlussschrift mit Schreibernennung und Datierung: Explicit Racionale divinorum anno domini Mo CCCCo LXV die 3o 24 mensis Septembris in Willanders per me Laurencium Smyt (24. September 1465). (445rb–447v) leer. (Ir–IIIv) leer. A. P. / U. S. 290 Cod. 578 Cod. 578 RAPHAEL VOLGOSIUS Pap. I, 10, I* Bl. 295 × 210. Brixen, 1480. B: Erstes bzw. letztes Blatt verschmutzt. Am rechten Blattrand Wasserschaden. Bilder und Nachweise der Was serzeichen s. WZMA. Wasserzeichnung mit Datierung übereinstimmend. Lage: VII,10,I. Am oberen Blatt rand Foliierung der beschriebenen Blätter 19. Jh. durch die ULB Tirol. S: Schriftraum mittig längs durch eine Blindlinie unterteilt: zwei Spalten mit wechselnder Zeilenzahl (40–51). Flüchtige Bastarda, lokalisiert und dat. Brixen, 1480 (Bl. 10vb). E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Am VD Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Bl. Ir Kaufvermerk 15. Jh. Helene 1481 lib XXXVI crucif VIII. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] C […] (radiert, Tinte), II3 B 16 (Bleistift) und 578 (Tinte). Am VD gelbes Signaturschild der ULB Tirol 578. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm 109. – Wretschko–Sprung 22. – Dolezalek II o. S. – Peintner, Stiftsbibliothek 122. – Katalog ULB Tirol VI 296, 382. – Krämer, Scriptores, s. v. „Volgosius, Raphael“ (Schreiber nicht identisch mit Raphael Volgosius, gest. 1427 in Padua. Freundlicher Hinweis Leonardo Granata, Padua, vgl. C. Bukowska Gorgoni, „Raffaele Fulgosio“. Dizionario Biografico degli Italiani L. Roma 1998, https://www.treccani.it/enciclopedia/ raffaele-fulgosio_(Dizionario-Biografico)/. (Ir) Tit.: Tituli omnium librorum legalium. (Ir) Kaufvermerk (s. bei G). (Iv) leer. (1ra–10vb) Raphael Volgosius: Alphabetisches Titelverzeichnis zum Römischen Recht, in der Hs. geordnet nach Institutionum, Digesti veteris, Digesti novi, Inforciati, Codicis, Authenticorum, Trium librorum codices ffeudorum. Institutionum. 1 Adoptionibus. Altiliano tutore. Auctoritate tutoris … vasallo qui contra constitutio nem / vocatis feudum. (10vb) Schlussschrift mit Lokalisierung und Datierung: Expliciunt rubrice iuris civilis secundum ordi nem alphabeti a Raphaele Volgosio compilate 1480 in Brixina. (Ir) leer. (Iv) Schuldvermerk: Item Hanns doctor Grendner ist mir schuldig […] gar IIII gulden minder IIII gulden. C. S. Cod. 579 291 Cod. 579 PUBLIUS OVIDIUS NASO Pap. 14 Bl. 300 × 210. Wiener Raum oder Tirol (?), 1491. B: Pap. bes. am Beginn und am Ende stark verschmutzt, am oberen rechten Blattrand fleckig (Wasserschaden), Bl. 12v–14r Tintenfleck, erstes und letztes Blatt an den Rändern eingerissen bzw. beschädigt, Bl. 2 Riss ge klebt. Nachweise und Bilder der Wasserzeichen s. WZMA. Lage: VII14. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Am Spiegel des VD Reste eines orangen Papiersignakels der ULB Tirol. S: Schriftraum 260 × 100/105, von jeweils zweifachen Blindlinien begrenzt, zu 30 Zeilen. Bastarda, dat. 3. November 1491 (Bl. 14v). Interlinear- und Randglossen von zwei zeitgenössischen Händen in Bastarda. Bl. 14v Zusätze von späterer Hand. A: Vereinzelt rote Auszeichnungsstriche, Paragraphzeichen, Unter- sowie Durchstreichungen, vereinzelt Rand notizen, Datierung (Bl. 14v) und Überschrift. Bl. 12v rote Zeigehand. Zweizeilige rote Lombarden. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Rücken stark beschädigt, Bindung gelöst. Am Rücken bzw. VD Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Rahmung, Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Bl. 1r Besitzvermerk Anfang 16. Jh. Magistri Nicolaus Leopoldi [ex] Insprugga XXXIIII (?, beschnitten) (vgl. Cod. 167; 1511 Lehrer an der Pfarrschule von St. Jakob, Innsbruck [W. Steiner, Die Lateinschule der St. Jakobspfarrkirche in Innsbruck, in: K. Drexel, M. Fink (Hrsg.), Musikgeschichte Tirols I: Von den Anfän gen bis zur Frühen Neuzeit (Schlern-Schriften 315). Innsbruck 2001, 404, 501], empfängt 1513 in Brixen die niederen Weihen und erhält 1515 Kanonikat, gest. 1535 [K. Wolfsgruber, Das Brixner Domkapitel in seiner persönlichen Zusammensetzung in der Neuzeit 1500–1803 (Schlern‑Schriften 80). Innsbruck 1951, 42 und 170; W. Senn, Musik und Theater am Hof zu Innsbruck. Geschichte der Hofkapelle vom 15. Jahrhundert bis zu deren Aufhebung im Jahre 1748. Innsbruck 1954, 13 und 46]). Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II1 D 3 (Tinte, radiert), II3 B 17 (Bleistift) und 579 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 579. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: J. Lechner, De codice Aenipontano 579 quo continetur Ovidi Remedia amoris. Commentationes Aeniponta nae 4 (1909) 17–104. – Kristeller I 439. – Neuhauser, Neustift 84. – Katalog ULB Tirol VI 296, 382. – Stefa ni 112. (1r–14v) Publius Ovidius Naso: Remedia amoris (Ed. E. J. Kenney. Oxonii 21995). Tit.: Publii Ovidii Nasonis Sulmonensis remedium amoris etc. Mit zahlreichen Interlinear- und Marginalglossen (Ed. Lechner 23–54). (1r, am Rand) ‹Antonius Volscus›: Vita Ovidii (Druck: u. a. GW M28918). Inc.: Publii Ovidii Nasonis poete vetusta equestris ordinis familia fuit … – Expl.: … ut tradit Eusebius. (1r–7v) V. 1–396. – (7v–14v) V. 397–814. Tit.: Publii Ovidii Nasonis liber secundus de remedio amoris. (14v) Schlussschrift mit Lokalisierung und Datierung: Τελος. (gestrichen: III Nonas Aprilis M CCCC 9o1). (rot:) In Lipcks (Lipsiae?) III Nonas (gestrichen: Aprile) Novembres M CCCC 9o1 (3. November 1491). Darunter Notiz von anderer Hand: Roma vetus veteres cur (!) te rexere Quirites (Walther, Initia Nr. 16879). (14v, am linken Blattrand, beschnitten sowie auf dem Kopf stehend) Federproben mit Briefanreden 16. Jh. von anderer Hand: 1) Freuntlicher lieber sorg auff ewr verschreyben so ir mir than habt fueg ich euch zw wissen das mir solichs. 2) Freuntlicher lieber vatter und schwager ich las euch … 3) Freuntlicher lieber schwager auff ewr verschr[ G. K. / A. P. 292 Cod. 597 Cod. 597 SERMONES. AUGUSTINUS FAVARONI Pap. 58 Bl. 270 × 210. Wiener Raum, um 1464/65. B: Pap. fleckig. Buchblock stark beschnitten. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Verwandte Wasserzeichen: u. a. Klosterneuburg, Stiftsbibl., CCl 308, T. I, CCl 561 (dat. 1465), CCl 743B. Lagen: 4.VI48 + (VI–2)58. Gegenbl. zu Bl. 49 und 50 herausgeschnitten (Textverlust). In den Lagenmitten Pergamentfälze, in der dritten Lage beschrieben (Ebermunt qui Penno. Eckehart et [?] frater Adili[…?] eundem / […]). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 225/230 × 150/155. Zwei Spalten, von feinen Tintenlinien begrenzt, zu 47–51 Zeilen. Bastarda von mehreren Händen: 1) Bl. 1ra–2vb, Z. 31, Bl. 3rb–vb; 2) Bl. 2vb, Z. 31–3ra; 3) Bl. 4ra–57vb; 4) Bl. 58ra– vb. Notizen am Rand (tw. stark beschnitten) von zeitgenössischer bzw. späterer Hand. Bl. 1r Vermerk 19. Jh. Alcorani doctrina. A: Bl. 1r rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen und Rahmung sowie zweizeilige einfache rote Lombarde. Ab Bl. 1v keine Rubrizierung, Platz für zwei- und dreizeilige Initialen freigelassen, nicht ausgeführt. Repräsentanten. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Pap. am VD unten beschädigt, seitlich eingerissen, am HD löchrig. Rücken oben tw. freiliegend, Signatur schild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] (radiert, Tinte), II3 C 18 (Bleistift) und 597 (Tinte). Am Rücken bzw. am VD gelbes Signaturschild der ULB Tirol 597. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Zumkeller Nr. 162. – Katalog ULB Tirol VI 373, 382. 1 (1ra–43va) Quadragesimale ‹De schola Christi› (48 Predigten) (vgl. J. B. Schneyer, Wegweiser zu lateinischen Predigtreihen des Mittelalters [Bayerische Akademie der Wissenschaften, Veröffentlichungen der Kommission für die Herausgabe ungedruckter Texte aus der mittelalter lichen Geisteswelt 1]. München 1965, 116 [ungedruckt]). (1ra) Inc.: Cum ieiunatis nolite fieri sicut ypocrite tristes Mt. VI (16). Pro huius thematis intro duccione invenio tres scolas. Prima igitur scola fuit Moisy in esse figurali … – Expl.: … ut Deus qui veritas est ut tibi in presenti ministret graciam et gloriam in futuris. (2va) Inc.: [S]icut credidisti fiat tibi Mt. 8 (13). Pro huius thematis introduccione invenio tres scholas scilicet wlgarium wlgarium (!), naturalium, moralium. Prima mensuratur experiencia secunda racione 3a (gestrichen: disar) discrecione … – Expl.: … et hereditatem eternam cum omnibus sanctis gloriose collocat et vitam eternam donat. Quam etc. (4ra) Inc.: [D]iligite inimicos vestros Mt. 5o (44). Pro huius thematis introduccione tres scolas reperio scilicet nature, scripture, gracie. Primo dico quod invenio scolam nature … – Expl.: … propter meritum quod expectas quod omnia pondera facit esse levia ad quod meritum nos per ducat Amen. (5rb) Inc.: [E]go sum nolite timere Mar. 6o (50). Pro huius thematis introduccione tres scolas reperio scilicet Epicuriorum, Platonicorum, Ypotheticorum. Sed in omnibus istis scolis doctrina legitur que secundum aliquam partem merito nobis timorem … – Expl.: … adimpletur hic per graciam et in futuro per gloriam Amen. (6rb) Inc.: [E]cce nunc tempus acceptabile ecce nunc dies salutis 2a Cor 6o (2). Pro huius thema tis introduccione tres scolas reperio et fortune, nature, racionalis creature. Prima scola scilicet nature dicit quod omnia naturalia … – Expl.: … sanantur anime vulnera hic in presenti etc. Cod. 597 293 (7ra) Inc.: [I]psi soli servies Mt. 4o (10). Pro huius thematis introduccione invenio tres scolas scilicet Heremitarum, Predicatorum, Minorum. Prima scola est Heremitarum in Egidio qui videtur velle tenere quod si primus homo non peccasset … – Expl.: … quod merito nostrum servicium … cum nobis hic in presenti per graciam et in futuro per gloriam. Amen. (8rb) Inc.: [C]um venerit filius hominis in maiestate sua etc. Mt. 26 (recte: 25,31). Pro huius thematis introductione invenio tres scolas scilicet Hebraicam Grecam et Latinam. Prima scola est Hebraica in qua considero Salomonem facientem iusticiam … – Expl.: … ibunt miseri in supplicium eternum, iusti autem in vitam eternam. Quam nobis concedat etc. Amen. (9va) Inc.: [D]omus mea domus oracionis vocabitur Mt. 21 (13). Pro huius thematis introduccio ne invenio tres scolas scilicet Belzebub in Babilonia, Iani in Roma, Phariseorum in Ierusalem. Prima scola fuit in magna civitate Babilonie in qua tractabatur … – Expl.: … que non licent homini loqui et postea ait Cupio dissolvi et esse cum Christo in gloria quam nobis concedat etc. Amen. (10vb) Inc.: [S]ignum non dabitur ei Mathei 12 (39). Pro huius thematis introduccione invenio tres scolas scilicet mechanicas, trepidas et incredulas. Primo invenio scolas mechanicas et iste sine dubio ut earum opera cognoscantur … – Expl.: … rem sacramenti. Ad quod nos perducat qui cum patre et spiritu sancto vivit etc. Amen. (12ra) Inc.: [M]iserere mei fili David Mt. (gestrichen: XIo) XV (22). Pro huius thematis intro duccione tres scolas reperio scilicet illustrium magnificorum, 2a eloquencium, 3a decipiencium et proditorum. Prima scola fuit illustrium etc. … – Expl.: … si nos dabimus intercedente merito recipiemus gloriam. Ad quam ille nos perducat etc. (13rb) Inc.: [S]anus fiebat a quacumque detinebatur infirmitate Ioh. Vo (4). Pro huius thematis introduccione invenio tres scolas videlicet phisicam Avicenne, cyrusicam Galieni, ewangelicam Ihesu Christi. In prima scola fuit magnus medicus naturalis et in illa legebatur … – Expl.: … sanati sumus hic in presenti per graciam et in futuro per gloriam Amen. (14rb) Inc.: [M]iserere nostri Deus omnium et respice nos Sapiencie 28 (recte: Sir 36,1). Pro huius thematis introduccione tres scolas reperio ethicam seu monasticam, yconomicam seu fami liarem, politicam seu civilem. Prima scola est monastica in qua legitur … – Expl.: … misericor diam tuam nos redemisti et viam glorie nobis aperuisti hic per graciam et in futuro per gloriam. Ad quam nos etc. (16ra) Inc.: [S]ic ambuletis ut habundetis magis prima ad Thessol. (!) 4o (1). Pro huius thematis introduccione invenio tres scolas. Prima scola est agricultorum qui habundat in gracia tempora li cum tales fructus terre et graciam Dei … – Expl.: … Quod apostolus considerans dicit in fine epistule Non enim vocavit nos Deus in immundiciam sed in sanctificacionem hic per graciam etc. Amen. (17rb) Inc.: [T]ransfiguratus est ante eos Mt. XVII (2). Pro huius thematis introduccione invenio tres scolas scilicet alchimistarum, artistarum, poetarum. In omnibus autem hiis scolis legitur transfiguracio. Prima scola est alchimistarum … – Expl.: … eius cantu Ecce quanta est paradisi gloria. Ad quam nos perducat Amen. (18vb) Inc.: [I]n peccato vestro moriemini Io. 8 (21). Pro huius thematis introduccione invenio 3 scolas scilicet celi, paradisi, mundi. Prima ergo scola fuit celi. Nam in principio creavit Deus celum et terram … – Expl.: … septem sunt peccata que impedire querunt animam ne valeat ad gloriam paradisi pervenire promissam in lege. Ad quam gloriam nos etc. Amen. (20ra) Inc.: [Q]ui se exaltat humiliabiliter Mt. 23 (12). Pro huius thematis introduccione invenio tres scolas scilicet elegancie in naturalibus affluencie in temporalibus eminencie in dignita tibus … – Expl.: … paciens pauper factus est ut ad pacienciam superbum reduceret hic in p resenti per graciam etc. (21rb) Inc.: [N]escitis quid petatis Mathei 4o (recte: 20,22). Pro huius thematis introduccione tres scolas reperio videlicet mundi, carnis, demonis. Prima scola est mundi in 1a legitur apparencia 294 Cod. 597 sive existencia … – Expl.: … et induamur arma lucis sciencie et gracie. Quam nobis concedat qui sine fine vivit et regnat Amen. (22va) Inc.: [M]ortuus est dives et sepultus est in inferno Luc. 16 (22). Pro huius thematis intro duccione invenio tres scolas scilicet Sardanapelli in Asya, Catiline in Roma et Attile in Ungaria. Prima ergo scola est Sardanapelli … – Expl.: … contemptus Dei inhumanitas et oblivio mortis procedas sicut scis quia amplius de 1o sermone non inveni in exemplari. (24rb) Inc.: [H]eres est. Venite occidamus eum Mt. 21 (38). Pro huius thematis introduccione tres scolas invenio scilicet Polestinoris, Mesabellis, Pignaleonis. Prima scola fuit Polestinoris qui fuit rex cuiusdam insule … – Expl.: … et vineam suam locabit aliis agricolis quia ibi modo dicun tur esse Enoch et Helyas. Rogemus ergo etc. (25va) Inc.: [D]issipavit bona sua vivendo luxuriose Luce III (recte: 15,13). Pro huius thematis introduccione tres invenio scolas scilicet Medee, Dyodonis, Ysiphile. Prima ergo scola fuit Medee nam Medea fuit filia Ochis (!) regis qui regnavit in insula Cholchos (!) … – Expl.: … sepe recommendemus nos virgini Marie qui impetrabit graciam pro nobis hic in presenti et in futuro gloriam. Ad quam etc. (27ra) Inc.: [E]stote imitatores Dei sicut filii carissimi (Eph 5,1). Pro huius thematis intro duccione invenio tres scolas scilicet dominorum, generatorum, magistrorum. Prima scola fuit dominorum et in ista et in aliis legitur quod omnes intendunt sibi assimilare … – Expl.: … nos ad bonum vocant sed pocius repellunt et Christus semper stat ad hostium et dicit Qui me invenerit inveniet vitam hic per graciam et in futuro per gloriam Amen. (28ra) Inc.: [O]mne regnum in se divisum desolabitur Luce XI (17). Pro huius thematis intro duccione invenio tres scolas scilicet Thebanam, Troyanam, Romanam. Prima scola fuit Thebana et in hac pro sui tempore lecta fuit divisio … – Expl.: … castri cum muris circumdata cum duobus custodibus cum portis cum populo etc. Procedas sicut scis quia ulterius non invenio. (29ra) Inc.: [N]emo propheta acceptus est in patria sua Luce 4o (24). Pro huius thematis intro duccione tres scolas invenio scilicet filiorum, inimicorum, discipulorum. Prima scola est filiorum. In hac considero inter filios primi parentis … – Expl.: … invidia originetur. Quoniam exercitetur in quo figuretur, procedas sicut scis et vis etc. (29rb) Inc.: [Q]uecumque solveris super terram erunt soluta et in celo Mt. 18 (18). Pro huius thematis introduccione tres scolas invenio scilicet nuptiarum, victoriarum et scienciarum. Prima ergo fuit nuptiarum et illam invenio in rege Aswero … – Expl.: … auctoritas originetur. Quoniam exercitetur in quo figuretur. Et procedas sicut placet etc. (29vb) Inc.: [Q]ue procedunt de ore hec coinquinant hominem Mt. VIo (recte: 15,18). Pro huius thematis introduccione invenio tres scolas invenio (!) Kathonis, Tulii et Senece. Prima scola est Cathonis et quamquam intra illam bona vita legatur tamen inter alia qui ibi leguntur … – Expl.: … verbum originetur. Quoniam exercitetur in quo figuretur dicas de verbo mentali et vocali et scripto etc. (30rb) Inc.: [I]mperavit febri et dimisit illam Luc. 4o (39). Pro huius thematis introduccione invenio tres scolas videlicet Origenis, Augustini, Ieronimi. Prima scola est Origenis cuius talis fuit opinio seu doctrina … – Expl.: … procedas qualiter originatur ex proporcione naturalium electorum seu 7 qualitatum etc. (30va) Inc.: [H]ic est vere salvator mundi Io. 9 (recte: 4,42). Pro huius thematis introduccione invenio tres scolas scilicet lapidum, herbarum, verborum. Prima ergo scola est lapidum et in illa legitur magna virtus. Nam videmus quod carbunculus … – Expl.: … ut hoc facto possit haurire graciam hic in presenti et in futuro gloriam Amen. (32ra) Inc.: [N]ec ego te contempnabo (!) Ioh. 8 (11). Pro huius thematis introduccione invenio tres scole (!) scilicet wlgaricam, canonicam, theologicam et in hiis legitur quod nullus possit iudicari … – Expl.: … unus sit iudex et quod iudex sine misericordia est severus. Procedas sicut tu scis. Cod. 597 295 (32rb) Inc.: [N]on sumus filii ancille sed libere Gal. 6o (recte: 4,31). Pro huius thematis intro duccione tres scolas invenio scilicet potenciarum, naturarum, gramaticorum. Nam prima est potenciarum et quia habemus plures potencias … – Expl.: … unde filii originentur. Quoniam exercitentur in quo figurentur procedas sicut scis et vis et nota libri falsitatem. (32vb) Inc.: [D]istribuit discumbentibus Io. 6o (11). Pro huius thematis introduccione reperio tres scolas mirificam, munificam, magnificam. Prima scola est mirificum (!) et hec scola est Dei. Prope enim Deus qui fecit mirabilia … – Expl.: … utrum plus mereatur dando pauperibus quam divitibus procedas sicut placet. (33rb) Inc.: [S]olvite templum hoc et in tribus diebus excitabo illud Io. 2o (19). Pro huius thema tis introduccione invenio 3 scolas scilicet extimam, intimam, extrinsecam. Prima scola dicitur extima et hec est excellencie dignitatis et in hac lego trinitatem … – Expl.: … unum solucio originetur. Quoniam exercitetur in quo figuretur procedas sicut scis et dic quod originatur a peccato etc. (33va) Inc.: [Q]uomodo hic litteras scit cum non didicerit Io. 7o (15). Pro huius thematis intro duccione invenio tres scolas scilicet organicam, scientificam, ewangelicam. Prima scola dicitur organica et in hac legitur quod quis doctus esse non potest … – Expl.: … utrum infinite videat facialem visionem divinitatis. Nunc procedas sicut tibi placet. (34ra) Inc.: [E]tatem habet ipse de se loquatur Io. 9 (21). Pro huius thematis introduccione tres scolas invenio scilicet examinacionis, confessionis, obligacionis. Prima scola est examinacionis et in ista scola legitur quod reus dato quod omnia sunt allegata et approbata … – Expl.: … non est opus valentibus medicus etc. sic peccator spiritualiter dum vivit etc. (34va) Inc.: [T]ibi dico surge Luce 7o (14). Pro huius thematis introduccione 3 scolas invenio scilicet equitatis, benignitatis, sanctitatis. Prima scola est equitatis et in hac legitur quod rele vetur infirmus … – Expl.: … sicut mundari ad maculatum fuisse et qualiter causatur a libero arbitrio etc. (34vb) Inc.: [D]omine si fuisses hic frater meus non fuisset mortuus (Io 11,21). Pro huius thema tis introduccione reperio tres scolas scilicet elementalem, spiritualem, iudicialem. In hiis tribus scolis legitur mors. Prima ergo scola est elementalis quia composita originaliter ab hiis cum omne compositum resolvi … – Expl.: … in sphera et in cursu eiusdem etc. (35rb) Inc.: [P]er proprium sanguinem introivit semel in sancta eterna redempcione inventa (Hbr 9,12). Pro huius thematis introduccione invenio tres scolas scilicet cordialem, pigmentalem, usualem. Prima est cordialis in qua legitur quod sanguis est summum elementum cordis … – Expl.: … Ideo Dei filius sanguinem suum posuit homine ut dicunt verba proposita. In quibus queras 3a etc. (35vb) Inc.: [M]ortem non gustabit in eternum Io. 9 (recte: 8,52). Pro huius thematis intro duccione invenio tres scolas scilicet planete, comete, monete. Prima est planete et in hac legitur vita secundum quod melior est constellacio vel planeta de tanto efficimur levioris vite … – Expl.: … In quibus tria quero scilicet unde nostra redempcio originetur, quoniam excitetur, in quo figu retur. Dicas sicut scis. (36ra) Inc.: [Q]ueritis me et non invenietis Io. 7mo (34). Pro huius thematis introduccione invenio tres scolas scilicet motorum, auditorum, bellatorum. Prima est motorum et in illa legitur lux gracie unde Gen. primo dixit Deus fiat lux et facta est lux … – Expl.: … et querat in scolis fidei quia eum querentibus dicitur verbum propositum. Queritis me etc. In quibus verbis quere 3a. Unum etc. (36va) Inc.: [N]on enim volebat in Iudeam ambulare quia querebant eum Iudei interficere (Io 7,1). Pro huius thematis introduccione 3 scolas invenio scilicet Martis, partis, mortis. Prima est Martis. Nam Mars ut pluries dictum est planeta malivolus est et nati sub illo sunt pessimi … – Expl.: … Non enim volebat etc. In quibus queras utrum velle interficere originetur et procedas ulterius sicut scis etc. 296 Cod. 597 (36vb) Inc.: [O]pera que ego facio in nomine patris mei ipsa testimonium perhibent de me Io. 20 (recte: 10,25). Pro huius thematis introduccione invenio tres scolas scilicet veneficas (darüber geschrieben: b), angelicas (darübergeschrieben: a), truffaticas. Prima scola est angelica et in hac legitur magna opera facta fuisse et primo legitur de archangelo Michaele … – Expl.: … et dicitur quod a virtutibus ut patet de pacientibus prosequerer sicut scis et sicut tibi placet. Sequitur. (37rb) Inc.: [L]acrimis rigavit pedes meos Luce 7o (44). Pro huius thematis introduccione invenio tres scolas scilicet arborum et plantarum, vaporum et nebularum, dolencium animarum. Prima scola est plantarum et arborum in qua talis doctrina legitur quod quando arbores plorant … – Expl.: … et sanguinem quo suscepto anima ad Deum evolavit et sic habuit graciam in presenti et in futuro gloriam Amen. (38rb) Inc.: [C]ollegerunt pontifices et Pharisei concilium adversus Ihesum Io. primo (recte: 11,47). Pro huius thematis introduccione invenio tres scolas scilicet urbis fundatorum, in campo bellatorum, in campo clericorum. Prima scola est urbis fundatorum … – Expl.: … illi qui hic dat graciam et in futuro gloriam. Ad quam nos perducat idem dominus benedicans in secula Amen. (39va) Inc.: [P]ater clarifica nomen tuum Io. 12 (28). Pro huius thematis introduccione invenio tres scolas scilicet sapiencie, presencie, refulgencie. Prima scola est sapiencie in qua legitur quod sapiencium nomen clarificatur per famam in multis locis … – Expl.: … quero in quo n omine Dei figuretur. Respondeo quod figuratur in nomine generali procedas sicut vis. (40va) Inc.: [N]on rapinam arbitratus est esse se equalem Deo ad Phil. 2o (6). Pro huius t hematis introduccione invenio 3 scole (!) angelice nature, humane creature, artificialis figure (gestrichen: Pro huius thematis introduccione) et in omnibus hiis legitur forma rapine. Prima scola est an gelice nature in qua legitur rapina … – Expl.: … dico quod originatur ex inordinato appetitu et disproporcionato dicas sicut scis. (40vb) Inc.: [C]eperunt ramos palmarum et processerunt obviam ei Luce 12 (recte: Io 12, 13). Pro huius thematis introduccione invenio tres scilicet olive in Minerba, lauri in Mercurio, palme in Marte. Prima scola est olive in Minerba. Nam ut possum capere ex dictis poetarum oliva consecrata est in nomine Minerbe … – Expl.: … et tunc dominus recipitur in Iherusalem id est in visionem pacis ad quam nos perducat Amen. (42ra) Inc.: [M]artha ministrabat Io. 12 (2). Pro huius thematis introduccione invenio tres scolas scilicet internas, alternas, subalternas. Prima est interna et est voluntatis et in hac legitur quod ea que sunt in homine ipsi voluntati principaliori et digniori subalternas serviunt … – Expl.: … recipiat condimentum hoc per graciam et in futuro per gloriam Amen. (43ra) Inc.: [V]enite mittamus lignum in panem eius Iere. 41 (recte: 11,19). Pro huius thematis introduccione invenio tres scolas scilicet mechanicas, piraticas, maleficas. Prima scola est me chanica et hec utitur ligno … – Expl.: … si crux que est lignum poneretur in membris corporis Christi Amen. 2 (43va–57vb) Augustinus Favaroni: Sermones bzw. Interrogationes seu introductiones quadrage simales (vgl. Schneyer II: Augustinus [Favaroni] de Roma, München, BSB, Clm 4706, Bl. 1–36: Sermones 43 und 50‑52 in Hs. nicht enthalten. Zumkeller Nr. 162. W. Eckermann, „Augustinus Favaroni“. Theologische Realenzyklopädie IV. Berlin 1979, 739–742). Inc.: [R]edde illis mercedem Mt. 20 (8). Questio inter doctores scolasticos solet moveri utrum Deus posset non retribuere mercedem summe beatitudinis creature racionali diligenti eum fide liter … – Expl.: … et hoc in ligno sanctissime crucis de cuius morte et passione hodie ecclesia facit specialem mencionem etc. Enthält 49 Sermones, Sermo 3 im Druck nicht enthalten: (44ra) Inc.: [V]obis datum est nosse misterium regni Dei Mt. XIIIo (11). Questio oritur inter doctores utrum in hac vita possit (gestri chen: aliquis) Deum videre facialiter … – Expl.: … doctores scolastici pro maiori parte tenent cum Augustino in hac questione etc. Cod. 605 297 Schlussschrift: Expliciunt distinctiones reverendissimi magistri Augustini de Roma ordinis Here mitarum sancti Augustini. (57ra–vb) Auflistung der Sermones, jeweils als Quaestio formuliert (ohne Nummerierung, ohne Blattangabe). Inc.: [U]trum Deus posset non retribuere mercedem summe beatitudinis creature racionali di ligenti eum fideliter et perseveranter … – Expl.: … Utrum aliqua pura creatura potuerit esse sufficiens ad redimendum genus humanum. 3 (57vb) Auctoritates-Zitate zu De Anima. Tit.: Nota de anima. – Inc.: Cassiodorus in libro De anima. Magistri scolarium (!) litterarum aiunt animam esse substanciam simplicem speciem naturalem distantem a materia corporis sui organum membrorum … – Expl.: … sicut dicitur in libro de motu cordis. Item Seneca Anima est spiritus intellectualis ad beatitudinem in se et in corpore ordinatus. 4 (58ra–vb) Sermo, unvollständig. Inc.: [V]os estis (darüber: autem) genus electum regale sacerdocium gens sancta etc. 1. Pe. 2 (9). Hec verba sunt beati Petri ad commendacionem et dignitatem sacerdotalem. In verbis premissis possumus notare triplicem gradum sive excellenciam cuiuslibet sacerdotis. Primo eius dignita tem … – Expl. mut.: … unico verbo liberare posset (gestrichen: ut) carcer aperiretur (gestrichen: et) vincula dirumperentur et liber a capti [ Tw. Sätze durchgestrichen, Ergänzungen und Zusätze am Blattrand (beschnitten). U. S. Cod. 605 PETRUS HISPANUS. DE MODIS SIGNIFICANDI Perg. I, 8, I Bl. 185 × 135. Entstehungsort unbestimmt, Ende 13./Anfang 14. Jh. B: Bl. 1 stark fleckig, Bl. 5 Nahtspur, vereinzelt kleine Löcher. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + I2 + III8 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Zw. Bl. 1 und 2 sowie nach Bl. 2 und 8 Textverlust: urspr. wohl Quaternionen. Am oberen Blat trand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 140/145 × 105. Zwei Spalten, von nur mehr schwach erkennbaren Tintenlinien gerahmt, zu 35 Zeilen auf schwacher Tintenlinierung. Zirkelstiche. Textualis. Am Blattrand vereinzelt Nachträge von zeit genössischer Hand. A: Vereinzelt rote Auszeichnungsstriche, Paragraphzeichen, Zeilenfüllsel, Über- und Schlussschrift sowie Schreiberverse. Bl. 2rb Ergänzung am Blattrand in blauer Tinte. Zweizeilige rote Lombarden mit Punktver dickungen. Bl. 5va achtzeilige rote Initiale mit verzierten Ausläufern. Bl. 1ra sechszeilige abgeriebene Initiale, mit blauen und roten Umrisslinien nachgezeichnet, linker Buchstabenschaft abgetreppt. Bl. 5v und 6v spätere Bleistiftkritzeleien. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Pap. tw. abgerieben und beschädigt. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, drei Bünde, von jeweils dreifachen Streicheisenlinien gesäumt. Im zweiten Feld Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neu stift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II`6 D 13 (? radiert, Tinte), II`3 C 5 S. 2 (Bleistift) und 605 (Tinte). Am Rücken bzw. VD gelbes Signaturschild der ULB Tirol 605. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Triden tina 20. Jh. 298 L: Cod. 605 Kristeller I 439. – Katalog ULB Tirol VII 43, 484. (Ir) leer bis auf Bleistiftnotiz: Balos v. Menter (?). (Iv) leer. 1 (1ra–5rb) ‹Petrus Hispanus›: Tractatus sive Summulae logicales, unvollständig (Ed.: L. M. de Rijk, Tractatus called afterwards Summule logicales [Philosophical texts and studies 22]. Assen 1972). Gegenüber Ed. tw. erweitert, tw. leicht abweichend. (1ra–vb) Tract. 1: De introductionibus, unvollständig. Inc.: Dyaletica (!) est ars arcium sciencia scienciarum ad omnia methodorum principia habens viam. Sola enim dyalectica probabiliter disputat de principiis omnium artium … – Expl. mut. (cap. 8): … ut sortes currit [ (2ra–vb) Tract. 3: De praedicamentis, unvollständig, Tit. Bl. 2rb nachgetragen. Inc. mut. (cap. 2): ] quedam forma substancialis ut anima est substancialis forma hominis alia autem et forma accidentalis ut album homini. Prima harum … – Expl. mut. (cap. 12): … et quibus dam aliis. Item substancia non sus[ (3ra–5rb) Tract. 5: De locis, unvollständig. Inc. von Ed. abweichend. Inc. mut. (cap. 20): ]nens et materia transiens. Permanens est ut in cultello ferrum transiens est ut in pane farina et in vitro … – Expl. (cap. 40, Schluss von Tract. 5): … argumentum construc tive et destructive. (5rb) Schlussschrift: Explicit tractatus. Darüber und darunter jeweils ein Schreiberspruch: 1) Lauda scriptorem donec videas meliorem (Colophons Nr. 22151. Vgl. Walther, Proverbia Nr. 13516a). 2) Detur pro penna scriptori pulchra puella (Colophons Nr. 20587. Walther, Proverbia Nr. 5527a). Lit.: M. Grabmann, Handschriftliche Forschungen und Funde zu den philosophischen Schriften des Petrus Hispanus, des späteren Papstes Johannes XXI. (SB der phil.-hist. Abt. der Bayerischen Akademie der Wissenschaften 9). München 1936, 1–85. Zur Verfasserfrage vgl. Rijk IX–XXIV. 2 (5va–8vb) De modis significandi (Grammatica speculativa), unvollständig. Anonymer Traktat nach (tw. übereinstimmend mit) Thomas de Erfordia: De modis significandi (vgl. Ed.: M. F. García, Lexicon scholasticum philosophico-theologicum. Hildesheim 1974 [hier Iohannes Duns Scotus zugeschrieben], Pars 1, cap. 46–53. Vgl. auch G. L. Bursill-Hall, Grammatica speculativa of Thomas of Erfurt. London 1972, 276–310). Inc.: Ad cognoscendam naturam construccionis in se et in suis partibus sive speciebus sciendum quod construccio est quoddam incomplexum et quia incomplexum secundum Boecium cogno scitur dupliciter. Diffinicione que ordinatur ad cognoscendam rem in se … – Expl. mut.: … vel accusativus ut legitur librum vel ablati[ Lit.: S. Grotz, Thomas von Erfurt. Abhandlungen über die bedeutsamen Verhaltensweisen der Sprache (Tractatus de Modis significandi) (Bochumer Studien zur Philosophie 27). Amsterdam, Philadelphia 1988, VII–XLIV. (Ir–v) leer. A. P. / U. S. Cod. 606 299 Cod. 606 TRACTATUS DE CONFESSIONE Perg. I, 6, I Bl. 220 × 155. Entstehungsort unbestimmt, 14. Jh. B: Dickes Perg. Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + III6 + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Urspr. möglicherweise Quaternio (vor Bl. 1r und nach Bl. 6v Textverlust). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 170 × 110. Zwei Spalten, von Tintenlinien gerahmt, zu 34 Zeilen auf Tintenlinierung. Zirkel stiche. Textualis. Am Rand vereinzelt Ergänzungen und Korrekturen tw. von zeitgenössischer, tw. von späterer Hand. A: Rote Überschriften, Paragraphzeichen, Auszeichnungsstriche, Trennungsstriche und Unterstreichungen. Zweizeilige rote Lombarden, tw. mit Ausläufern. Bl. 1ra fünfzeilige rote Lombarde. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift, 2. H. 18. Jh. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, sechs dreifache Streicheisen linien. Am Rücken bzw. VD Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II`6 B 4 (radiert, Tinte), II`3 C 6 S. 2 (Bleistift) und 606 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 606. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Katalog ULB Tirol VII 43, 489. (Ir–v) leer. Tractatus de confessione, unvollständig (vgl. Bloomfield Nr. 1125 und Suppl. Nr. 1125. Hamesse Nr. 9778. Kaeppeli Nr. 3196). (1ra) Prolog. Inc. mut.: ] nugis et ioco ac fabulis vacantes. Nota ad honorem individue trinitatis … – Expl.: … duximus compilare in quo tractatu agere. (1ra) Inhaltsübersicht. Tit.: Capitula. – Inc.: Quomodo confessor consideret condiciones confiten tis … – Expl.: … De casibus pro quibus est ad superiorem recurrendum. (1rb–6vb) Text. Tit.: Primum capitulum quomodo confessor debet considerare vitam et condiciones confitentis. – Inc.: Primum capitulum est quomodo confessor debet considerare condiciones confiteri volentis. Ubi notandum quod confessor sedens ad confessiones audiendas si confiteri volentem non cognoscit querat ab eo an sit suus parrochianus … – Expl. mut. (cap. 16): … potest confessor inquire re modum qui accidit sibi dormiendo an vigilando an aliquo tactu suo vel [ Vgl. Wilten, Stiftsbibl., Cod. 32 03 07, Bl. 215ra. Lit.: Schulte GQ II 530, Nr. 11; P. Michaud-Quantin, Sommes de casuistique et manuels de confession au moyen âge (XII–XVI siècles). Analecta Medievalia Namurcensia 13 (1962) 85, Index. (Ir–v) leer. A. P. 300 Cod. 614 Cod. 614 JURISTISCHE SAMMELHANDSCHRIFT Pap. I, 290, I Bl. 210 × 145. Tirol, T. I: 7./8. Jz. 14. Jh, T. II: 1. Viertel 15. Jh. (u. a. 1419), T. III: 1. Jz. 15. Jh., T. IV: 4. Jz. 15. Jh. B: Pap. im Zuge der Restaurierung 20. Jh. (s. bei G) am äußersten Blattrand tw. ergänzt, Bl. 173–182 verstärkt, Löcher geklebt. Vier Teile: T. I (I, 1–149), T. II (150–269), T. III (270–283), T. IV (284–290, I). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. E: Bibliothekseinband: rot gefärbtes Leder über dicker Pappe. Rignano sull’Arno (Prov. Firenze), 20. Jh. Einband und Bindung im Zuge der Restaurierung erneuert. Kanten gerade. Kapitale mit gelblichem und grünem Garn umstochen. Rücken: drei einfache Bünde, im untersten Feld Signaturschild (s. bei G). Spiegel Pap. 20. Jh. Spiegel des VD leer, am Spiegel des HD Etikett der Restaurierwerkstätte (s. bei G). G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. 1809 anlässlich der vorübergehenden Auf hebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r alte Signaturen der ULB Tirol II […] (radiert, Tinte), darüber 614 (Tinte) sowie Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauer leihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh., dieser auch Bl. 2r. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro S. Maria del Rosano), kein Restaurierbericht erhalten. Bl. Ir Signatur 20. Jh. Ms. 614 (Bleistift), am Rücken Signaturschild 20. Jh. 614. L: Schulte GQ II 410. – Wilhelm I 136 und II 109. – Wretschko–Sprung 22. – Neuhauser, Neustift 88. – Stefani 113. – Katalog ULB Tirol VII 94, 484. – Krämer, Scriptores, s. v. „Weinkern, Albertus“. I (I, 1–149) B: Bl. 119 am oberen Blattrand beschnitten (ohne Textverlust). Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: (I–1)I (Vorsatzbl. 20. Jh.) + VIII16 + 2.IV32 + 9.VI140 + (VI–3)149. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. Die letzten drei Blätter der letzten Lage herausgeschnitten (ohne Textverlust). Zwei Kustodenzählungen in zeitgenössischen schwarzen Zif fern: 1) am Lagenende beginnend bei 3, die zweite Lage hierbei übersprungen, folglich dritte Lage Forts. mit 4; 2) am Lagenanfang beginnend bei 1, Lage 1–5 in der Mitte oben, ab Lage 6 unten. S: Schriftraum ca. 160/170 × 105/120, seitlich meist von Tintenlinien gerahmt, zu 35–47 Zeilen. Bastarda. Ver einzelt Ergänzungen und Korrekturen am Blattrand von derselben Hand. Oberlängen in der ersten Zeile vereinzelt gedehnt. A: Bl. 1r–58r, 93r–97r, 105r–110r, 117r–125r, 129r–147v rote Auszeichnungsstriche, tintenfarbene und tw. rot übermalte Paragraphzeichen. Bl. 1r–17v, 23v–27v, 28v–45r, 57r–58r, 105r–110v, 117r–125r, 129r–147v am oberen Blattrand Titel der jeweiligen Kapitel in Rot angeführt. Durchgehend rote Überschriften sowie ein- bis fünfzeilige einfache Lombarden, vereinzelt mit Punktverdickungen, Ausläufern und Aussparungen; Lombarden bisweilen nicht ausgeführt. Bl. 1r vierzeilige rote Lombarde Q mit Aussparungen und als Binnen feld- und Caudafüllung Horizontalreihen aus winzigen Bögen (in Rot und hellem Tintenbraun). Bl. 46v–56v, 59r–60r, 61r–64r, 65r–66v, 67v, 68v–69r, 71v, 72v, 76v, 78r, 79v–80r, 81r–v, 82v–83r, 84r, 85r, 86r–v, 87v, 88v–89r, 91r–v, 97v, 99r–100v, 101v, 102v, 103v–104v, 112r–116r, 125r, 126r und 128r keine Rubrizierung. 1 1.1 (Ir–v) leer bis auf Signatur (s. bei G). (1r–147v) Raimundus de Pennaforte: Summa. (1r–139r) Summa de paenitentia (= Summa, Lib. 1–3) (Ed.: X. Ochoa, A. Diez, S. Raimundus de Pennaforte, Summa de paenitentia [Universa Bibliotheca iuris I B]. Roma 1976. – Kaeppeli Nr. 3407. Bloomfield Nr. 4963, 5054). (1r) Prooemium. (1v) Lib. 1. – (40v) Lib. 2. – (68r) Lib. 3. (73v) nur Rahmung sowie zweifacher Hinweis in den Ecken in Rot vacat. Cod. 614 1.2 301 (139r–147v) Summa de matrimonio (= Summa, Lib. 4) (Ed.: X. Ochoa, A. Diez, S. Raimundus de Pennaforte [Universa Bibliotheca iuris I C]. Roma 1978. – Kaeppeli Nr. 3408. Bloomfield Nr. 4943). (147v) Schlussschrift: Explicit summa casuum (darunter kleiner und in schwarzer Tinte wieder holt). (148r–149r) leer. Lit.: Schulte GQ II 410–413; Kuttner 443–445. (149v) Federproben: In nomine domini Amen. Anno nativitatis eiusdem. – Summa Raymundi. – Lex est defuncta ubi iudicis est manus uncta (darunter in kleinerer Schrift nochmals wiederholt) (Walther, Proverbia Nr. 13695). II (150–269) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: 10.VI269. Am Lagenanfang und -ende Kustoden in zeitgenössischen roten arabischen Ziffern. Lagenreklamanten. S: Schriftraum 140/145 × 90/95, von Tintenlinien gerahmt, zu 18–24 Zeilen. Bastarda mit zunehmend breiter werdendem Duktus, dat. 12. Dezember 1419 (Bl. 261r). Schreiber: Albertus Weinkern (u. a. auch Brixen, Priesterseminarbibl., Cod. D.19, dat. 1421; München, BSB, Clm 8375, dat. 1418 in Innsbruck, vgl. Krämer, Scriptores; belegt auch als notarius publicus in Mairhofer 514 [Stift Wilten 1426]). Hervorhebungen am Rand von gleicher Hand, Korrekturen und Ergänzungen tw. von anderer Hand. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen und Rahmungen. Vereinzelt Zeigehände. Zwei- und dreizeilige rote Lombarden. Bl. 151r neunzeiliger roter I-Versal mit ausgesparter Herzblattranke. Repräsentanten. 2 (150r–261r) Processus iudiciarius (dictus Schweidnitz). (150r) Verworfener Text. Tit.: Incipit processus iudiciarius dictus Sweidnicz de causis delegatis et subdelegatis. – Inc.: In nomine domini Amen. Circa processum iudiciarium in causis delegatis et subdelegatis et ordinariis primo premittitur cum impetrator rescripti … – Expl. mut.: … in specturi mandatis vestris [ (150v) nur Rahmung. (151r–261r) Tit.: Incipit processus iudiciarius dictus Sweidnicz de causis delegatis et subdelega tis. – Inc.: In nomine domini Amen. Circa processum iudiciarium in causis delegatis … – Expl.: … Item oppone quod non est talis casus vel causa in quo vel in qua potuit appellare ut supra dictum est etc. Deo gracias gracias Deo Deo gracias etc. Schlussschrift mit Schreibernennung und Datierung: Explicit processus iudiciarius dictus Sweid nicz de causis delegatis et subdelegatis. Qui scriptus est per Albertum Weinkernem notarium publicum ac clericum Frisingensis dioecesis anno etc. XIXo in vigilia sancte Lucie virginis etc. (12. Dezember 1419). (261v–269v) nur Rahmung. Parallelüberlieferung: u. a. Augsburg, Staats- und Stadtbibl., 2o Cod. 260a, Bl. 1r und 2o Cod. 312, Bl. 198ra. In Augsburg, Staats- und Stadtbibl., 2o Cod. 312 als Wiener Vorlesung des Magisters Nikolaus Schweidnitz überliefert. Zu Schweidnitz s. F. J. Worstbrock, „Schweidnitz, Nikolaus“. Verfasser lexikon2 VIII 929–930; P. Uiblein, Acta facultatis artium universitatis Vindobonensis 1385–1418 (Publikationen des Instituts für Österreichische Geschichtsforschung VI, 2). Graz u. a. 1968, 25, 35, 553. Vgl. auch Th. Muther, Zur Geschichte des Römisch-canonischen Prozesses in Deutschland während des vierzehnten und zu Anfang des fünfzehnten Jahrhunderts. Rostock 1872, 52–75. 302 Cod. 614 III (270–283) B: Bl. 283v stark verschmutzt. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzei chenbefund. Identische Wasserzeichen: Innsbruck, ULB Tirol, Cod. 588, T. II, Cod. 590, T. V [Klausen/ Diözese Brixen]). Lage: VII283. S: Bl. 270r–282r: Schriftraum 145/160 × 95/105, von Bleistiftlinien gerahmt, zu 27–31 Zeilen. Bastarda. Bl. 282v–283v: Schriftraum 180/185 × 100, von kaum sichtbaren Bleistift- bzw. Blindlinien seitlich begrenzt. Bastarda, Feder- und Tintenwechsel Bl. 282v, Z. 34. Von gleicher Hand auch Zusatz auf Bl. 282r. A: Bl. 270r–274v: hellrote Trennungspunkte, Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Rahmungen und Überschriften sowie zweizeilige Lombarden. Bl. 270r dreizeilige rote Lombarde mit un gelenkem tintenfarbenem Knospenfleuronnée im Binnenfeld. Bl. 275r–282r: vereinzelt hellrote Auszeichnungsstriche und Unterstreichungen, zahlreiche Paragraph zeichen und Überschriften sowie meist einzeilige Lombarden, beim Buchstabenwechsel zwei- bis vier zeilige Lombarden, tw. mit Punktverdickungen und minimalem Dekor. Bl. 282v–283v keine Rubrizierung. 3 (270r–273v) Iohannes de Deo: Concordantiae Decretalium et Decretorum (G. Murano, Initia operum iuris canonici medii aevi. A shortlist of works, arranged by their incipit words [https:// home.uni-leipzig.de/jurarom/manuscr/murano/initican.htm]). Tit.: Concordancie Decretalium et Decretorum. – Inc.: De constitucionibus d II. et III. per totum et d IIII c 1 … – Expl.: … Salomitane ubi Iohanne (!) plene notavit de hac materia. Lit.: Schulte GQ II 97–98. 4 (273v–274v) Erklärungen zur Gliederung des Decretum Gratiani, der Dekretalen und des Corpus iuris civilis. Inc.: Sciendum quod Decretum dividitur in tres partes principales, quarum prima vocatur Distin cciones … – Expl.: … siquis dogmata conswevit de hoc notari d III § I secundum Hugonem et Io. 5 (275r–282r) Regulae iuris civilis. Alphabetisch geordnete Sammlung von Rechtssprichwörtern und -regeln (A–U/V). Tit.: Regule iuris civilis huius littere A sunt hee. – Inc.: Alterius circumvencio alteri non prebet accionem. Actus legittimi ut supra. Aliud est vendere aliud vendicioni consentire … – Expl.: … furiosus absentis loco est et ita Pomponius L 1 epistolarum scripsit. Inc. wie Bamberg, Staatsbibl., Msc. Can. 44, Bl. 109r. (282r) Darunter: Ergänzungen von anderer Hand, nicht mehr alphabetisch geordnet. Inc.: Omnis res per quascumque causas nascitur per easdem et dissolvitur (Decretum Gratiani, p. 2, C. 27, qu. 2, cap. 4). Ea facta que dubia sunt ut quo animo in meliorem partem i nterpretremur (!) … – Expl.: … quis homagium compellatur. 6 (282v–283v) ‹Bonifatius VIII.›: Regulae iuris sexti libri Decretalium (CIC, ed. Friedberg II 1122–1124). Tit.: Regule iuris libri Sexti. – Inc.: Beneficium ecclesiasticum non potest licite sine institucione canonica obtineri … – Expl.: … qui verba legis complectens contra legis nititur voluntatem. Datum Rome apud sanctum Petrum quinto Nonas Marcii pontificatus nostri anno quarto. IV (284–290, I) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: (V–3)290 + 1I* (Nachsatzbl. 20. Jh.). Gegenbl. zu Bl. 284–286 herausgeschnitten. Gegenfalz zum Nachsatzbl. vor Bl. 284. S: Schriftraum 160/175 × 90/100, von Blindlinien gerahmt. Beschriftung vielfach über den gekennzeichneten Schriftraum hinausreichend. Bastarda (gleiche Hand wie Bl. 282v, Z. 34–Bl. 283v). 303 Cod. 617 7 (284r–289r) Summarium Quaestionum mercurialium Iohannis Andreae (vgl. Druck: u. a. GW 1735. G. Murano, Initia operum iuris canonici medii aevi. A shortlist of works, arranged by their incipit words [https://home.uni-leipzig.de/jurarom/manuscr/murano/initican.htm]). Tit.: Secuntur Questiones mercuriales d. Io. An. disputate super regulas iu. li. VI. – Inc.: Artat statutum doctores legentes et ut questiones disputatas … – Expl.: … An magistri composiciones librorum vel scriptorum sua nomina debeant in illis inscribere. 8 (289v) Bartholomaeus Brixiensis: Quaestiones dominicales, De conditionibus appositis, qu. 1, unvollständig (Druck: Köln 1570, 89). Tit.: Questiones dominicales Bartho. Brixiensis. – Inc.: Quidam habens uxorem condempnatus de crimine capitali … – Expl. mut.: … si reddendus sit an sit decapitandus [ Lit.: Schulte GQ II 86. (290r–Iv) leer. P. K. / U. S. Cod. 617 SAMMELHANDSCHRIFT Pap. I, 282 Bl. 220 × 145. Tirol (u. a. Neustift), T. I, II, IV: 7. Jz. 15. Jh. (u. a. 1467), T. III: 6. Jz. 15. Jh. (u. a. 1455) B: Vier Teile: I (I, 1–54), II (55–116), III (117–199[200]), IV (200[201]–281[282]). Am oberen Blattrand Foli ierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 187 doppelt angeführt. Neuzeitliche orange Papiersignakel der ULB Tirol. E: Gotischer Einband: rot gefärbtes Schafsleder über Holz mit Blindlinien. Tirol, 2. H. 15. Jh. VD und HD: von dreifachen Blindlinien gebildeter Rahmen, im Mittelfeld ein durch zweifache Linien ge bildeter innerer Rahmen, am VD dieser durch zweifache Linien diagonal in mehrere Rautenfelder unterteilt. Spuren von je fünf kleinen Beschlägen. Leder-Metall-Schließe. Am VD Titelschild (Perg.) Donatus misticus de octo partibus oracionis cum pluribus […] und Signatur (?) N 10. Kanten gerade. Rücken: Leder tw. be schädigt, zwei Fitzbünde und vier Doppelbünde, im ersten Feld Pergamentschild M: S: de Oratione devota, im zweiten Pergamentschild a (?) Sermones Festivales, im dritten Signaturschild (s. bei G). Am Spiegel des VD Pergamentmakulatur (s. bei Inhalt), Exlibris und Signaturen (s. bei G), am Spiegel des HD Reste urspr. aufgeklebter Pergament- und Papierblätter sowie Abklatsch. G: Besitzvermerke 15. Jh.: Bl. Ir Iste liber est monasterii s. Marie virginis ad gracias alias ad Novacellam dicti; Bl. 281v Iste liber est monasterii s. Marie in Novacella, darunter in Textura Iste liber est monasterii s. Marie in No. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] (radiert, Tinte), II`3 D 3 (Bleistift) sowie 617 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 617. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm II 108. – Kristeller I 439. – Kaeppeli Nr. 146. – Hamesse Nr. 5333. – Stefani 114–115. – Katalog ULB Tirol VII 102, 484. – Krämer, Scriptores, s. v. „Fresell, Jacobus“. I (I, 1–54) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Identische Wasserzeichen in T. IV. Lagen: 1I (Vorsatzbl.) + 4.VI48 + (VI–6)54. Gegenbl. zu Bl. 50–53 heraus geschnitten (ohne Textverlust). In den Lagenmitten Papierfälze. Gegenfalz zum Spiegel des VD sowie zu Bl. I zw. Lage 1 und 2. Lagensignaturen (a1, a2, …), tw. weggeschnitten. Lagenreklamanten. 304 Cod. 617 S: Schriftraum 140 × 90, von Blindlinien gerahmt, zu 30 Zeilen. Bastarda, dat. 1467 (Bl. 53r). Schreiber: Iaco bus Fresell de Scocia, vgl. Krämer, Scriptores. A: Rote Auszeichnungsstriche, Unterstreichungen und Überschriften sowie zweizeilige Lombarden. Repräsen tanten. 1 (VDS) Fragment eines Psalteriums: Ps 105,7–21. Schriftraum beschnitten, von Tintenlinien gerahmt, zu 23 Zeilen auf Zeilenlinierung. Textualis formata (Tex tura) Ende 13./Anfang 14. Jh. Rote Satzmajuskeln am Versbeginn. Mittig von Exlibris der Stiftsbibl. Neu stift überklebt. Inc. mut.: ] noster in universa terra iudicia eius … – Expl. mut.: … sue et principem omnis [ (Ir) Inhaltsverzeichnis und Besitzvermerk 15. Jh. (s. bei G). (Iv) leer. 2 (1r–53r) Donatus devotionis: De octo partibus orationis, Pars 1 (Druck: J. Rastell, Donatus de votionis de octo partibus orationis. London o. J. [um 1515]. – Bloomfield Nr. 8301 und 8731). (1r) Tit.: Incipiunt octo partes oracionis devote secundum ordinem Donati grammatici. (1r) Prolog. Inc.: Evangelica clamat hystoria. Sine intermissione orate hoc est semper quidem orat qui semper bene agit … – Expl.: … quod hec oracioni commendacio stet pro huius tractatus sequentis prologo et prohemio. (2r) Inhaltsübersicht. Tit.: Capitula prime partis huius Donati oracionis. (2v) Text. Inc.: Partes oracionis quot sunt? Octo. Que? Devocio lacrime dulcedo incendium ascensio meditacio contemplacio et languor. Devocio quid est … – Expl.: … et securitas eterne possessionis que ex eo et per eum habebuntur quam cognicionem eternaliter nobis concedat qui vivit et regnat in secula seculorum Amen. (53r) Datierung und Schreibernennung: Explicit (korr. zu Expliciunt) octo partes oracionis devo te scripte per Iacobum Fresell de Scocia primo die Iulii anno domini millesimo quadringente simo sexagesimo septimo (1. Juli 1467). Zur Überlieferung und zur Verfasserfrage s. A. I. Doyle, The European Circulation of Three Latin Spiritual Texts, in: A. J. Minnis (Hrsg.), Latin and Vernacular. Studies in Late-Medieval Texts and Manuscripts. Cambridge 1989, 129–146, hier v. a. 138–141. Vgl. auch Bibliothèque Natio nale, Catalogue Général des manuscrits latins VI. Paris 1975, 319–320 zu Paris, BN, Ms. lat. 3593, Bl. 1r; E. J. Devereux, Bibliography of John Rastell. Québec 1999, 95. (53v–54v) leer. II (55–116) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: 4.VI102 + (VI+2)116. Bl. 115 und 116 eingefügte Einzelbl. (mit Klebeband fixiert). Lagenreklamanten, tw. beschnit ten. In den Lagenmitten beschriebene Pergamentfälze. S: Schriftraum ca. 145 × 100, von Tintenlinien gerahmt, zu ca. 30 Zeilen. Bastarda. A: Rote Auszeichnungsstriche, Unterstreichungen, Überschriften sowie drei- und vierzeilige Lombarden mit einfachem Dekor. 3 (55r–62v) Tractatus quomodo religiosus proficere possit in religione (Hamesse Nr. 5333). (55r) Prolog. Inc.: Carissime frater. Vivatis in religione secundum doctrinam apostoli dicentis Sobrie et iuste et pie vivamus in hoc seculo expectantes beatam spem et adventum glorie magni Dei. In quibus verbis apostolus Paulus docet … – Expl.: … adventum glorie magni Dei non se habere credat per sentimenta et dulcedines sensibiles sed expectet. (55r–62v) Text. Inc.: De primo est sciendum duplicem esse ebrietatem vitandam … Secundo vo lens in religione proficere debet iuste cum fratribus vivere … – Expl.: … (Bl. 61r, mit Initiale letzter Abschnitt beginnend) Sobrie iuste et pie vivens ferventique zelo perseveranter et discrete Cod. 617 305 ad perfeccionem tendens nec tamen credens se certe ipsam habere … huius caritatis affectus magnus inflammatur ut tandem homo ad perfeccionem quam tendit perveniat et vitam eternam obtineat Amen. 4 (63r–116v) ‹Nicolaus Kempf de Argentina›: Tractatus de proponentibus religionis ingressum et de anno probationis (Hauréau, Initia App. I 107). (63r) Pars 1: 31 Kapitel. Tit.: Capitulum primum. – Inc.: Convertimini ad me et salvi eritis. Ysaie (45) 22. Quamvis divina miseracio multis variisque modis in scripturis sanctis omnis vocare dignata est … – Expl.: … qui suum propositum roboravit spiritu sancto et trinitate unus Deus in seculum benedictus Amen. Explicit prima pars huius tractatuli. (95v) Pars 2: 23 Kapitel. Inc.: Novicius indutus quantocius potest in generali confessione se expediat in qua non sit nimis scrupulosus … – Expl.: … Numquid iterum post veteres declinaciones mores et conswetudines. Deo Gracias. Zur reichen Parallelüberlieferung vgl. D. D. Martin, Kempf, „Nikolaus von Straßburg“. Ver fasserlexikon2 IV 1119. III (117–199[200]) B: Bl. 117 Riss. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Lagen: 7.VI199(200). In den Lagenmitten beschriebene Pergamentfälze. Lagenreklamanten. S: Schriftraum 165 × 120, von Tintenlinien gerahmt, zu 32 Zeilen. Bastarda, dat. 1455 (Bl. 134r und 191v [18. April]). Schreiber: Erasmus Lämpl (Bl. 134r und 191v), Chorherr in Neustift, vgl. Giner 10. A: Rote Auszeichnungsstriche, Unterstreichungen, Überschriften und zweizeilige Lombarden. Repräsentanten. 5 (117r–134r) Rabbi Samuel, ins Lat. übersetzt von Alfonsus Bonihominis: Epistola ad Rabbi Isaac (Kaeppeli Nr. 146 mit Angaben zur reichen Überlieferung, mit Nennung der Hs.). Abgeglichen mit GW M39838, gegenüber Druck tw. gekürzt, tw. leicht abweichend. (117r) Tit.: Incipit disputacio Iudeorum cum ipsis Christicolis. 26 Kapitel. (117r) Prolog. Inc.: Reverendissimo in Christo patri fratri Hugoni magistro ordinis fratrum Pre dicatorum dignissimo ac sacrosancte pagine professori. Eius subditus frater Alfonsus Bonihominis Yspanus promissam obedienciam … – Expl.: … per te Deo volente in veritate confirmari et in dubiis declarari. (117v) Text. Cap. 1–26. Inc.: Desidero domine mi certificari per te ex testimoniis legis et prophe tarum et aliarum scripturarum … – Expl.: … contra dictum et eius scripturam docuit ut corruptus homo penitus et ignorans. (134r) Schlussschrift mit Schreibernennung und Datierung: Explicit epistula magistri Samuelis missa ad Rabbi Ysaac magistrum sinagoge. Excerptus per Erasmum Lämpel in Ässnigk ( Assling?) plebanum anno 1455. 6 (134v–193v) Sermones de sanctis et de tempore, u. a. ‹Peregrinus Oppoliensis›: Sermones de sanctis (Schneyer 4, 559–570 [Nummern – wenn nicht anders angegeben – im Folgenden darauf Bezug nehmend]). Tit.: Sermones de sanctis. (134v) De s. Andrea (Nr. 196). Expl.: … et viginti milia hominum sua predicacione convertit. Ecce (?) quam viriliter etc. – (135r) De s. Nicolao (Nr. 198). (136r) Tit.: De sancto Thoma. – Inc.: Affer manum tuam in latus meum. Ioh. 20 (27). Benigni homines habent modum dum vident aliquem carum amicum cecidisse in aliquam passionem solent eos instruere … – Expl.: … sine mortalibus existentibus. Rogemus. 306 Cod. 617 Parallelüberlieferung: Neustift, Stiftsbibl., Cod. 78, Bl. 8vb (Entstehungsort: Raum Brixen [Neu stift?], 1. H. 15. Jh.); Innsbruck, ULB Tirol, Cod. 71, Bl. 21rb (Vorbesitzer: Wilten, Entstehungs ort: Tirol, 15. Jh.), Cod. 207, Bl. 5va (Vorbesitzer: Schnals, Entstehungsort: Österreich, dat. 1440, Schreiber: Christianus de Engelprecz). (136r) Tit.: In nativitate domini. – Inc.: Parvulus natus est nobis. Ys. (9,5). Circa nativitatem domini tria notantur. Primo natus describitur secundo nativitatis tempus tercio nativitatis utili tas. De primo sciendum quod fuit natus admirabilis ineffabilis formidabilis venerabilis … – Expl.: … faciat nos tanti gaudii participes quod nobis prestare dignetur. (136v) De s. Stephano (Nr. 204). – (137r) De s. Iohanne ap. (Nr. 207). Expl. (geringfügig erwei tert): … perseveravit in fide Christi ut vicissitudinem amicicie Christi proposse daret. Rogemus ergo (Expl. wie Neustift, Stiftsbibl., Cod. 78, Bl. 15vb). – (137v) De s. Innocentibus (Nr. 208). Expl.: … quia nomen patris eius scriptum est in frontibus eorum scilicet quod vocabuntur filii Dei in eternum. (138v) Tit.: In circumcisione domini. – Inc.: Circumcisionis dies est solemnis quia octavus nati vitatis Christi est. Voluit autem dominus die octava circumcidi unde Rabi Moyses philosophus et theologus licet Iudeus … – Expl.: … unde hec dies novus annus dicitur. (139r) Tit.: In epiphania domini. – Inc.: Cum natus esset Ihesus in diebus Herodis regis etc. (Mt 2,1). Hodie triplex festivitas celebratur. Una est de adoracione magorum … – Expl.: … sci licet virtutum revertamur ad penitenciam celestem Amen. (139v) In conversione s. Pauli (Nr. 216). Expl. (leicht abweichend): … quod lac fluxit loco san guinis de corpore suo. (Expl. wie Neustift, Stiftsbibl., Cod. 78, Bl. 23va). – (140r) In purifica tione BMV (Nr. 217). Expl. (leicht erweitert): … ipsa stat pacem et veniam obtinendo petimus et graciam impetrando. Rogemus etc. – (141r) In cathedra s. Petri (Nr. 220). Expl.: … et in Romana kathedra sedit annis XV. – (142r) De s. Matthia (Nr. 222). Expl.: … et ei devotissime servivit. (143r) Tit.: De commendacione ieiunii. – Inc.: Adest tibi venerabile tempus Quadragesime … – Expl.: … Qui se in paupere esurire testatur. Rogemus etc. (144r) De s. Gregorio (Nr. 126). Expl. (leicht abweichend): … Ultima felicitas quam recepit est visio Dei quam ipse recepit que in celis finitur ad quam perducat nos etc. – (145r) In annuntia tione BMV (Nr. 228). Expl.: … quis hominibus placeret. Respondit Si gesseris optima et locutus fueris pauca. Rogemus. (146v) Nr. 230. Inc. (geringfügig abweichend): Egredietur virga de ra dice Yesse. Ys. (11,1). Legitur de quinque virgis laudabilibus per quas beata virgo Maria merito figuratur … (147r) Nr. 229. Inc. (leicht abweichend): Fons crevit in flumen magnum. Hester VI (recte: 10,6). Ibi dic historiam qualiter Mardocheus sompniavit duos dracones … (148r) ‹Graeculus›: Sermo in annuntiatione BMV (Schneyer 2, 227, Nr. 267). Tit.: De eadem. (149r) Tit.: In die palmarum. – Inc.: Quadruplex agitur festum in hac septimana scilicet festum veneracionis festum recreacionis festum reconciliacionis … – Expl.: … Christus a morte resur gens discipulus ianuis clausis potuit apparere vel sicut virginis uterum ingrediens etc. (150r) ‹Leo OCist.›: Speculum corporis Christi sive Dialogus increduli et ecclesiae de sacra mento eucharistiae (Schneyer 4, 40, Nr. 369, hier ohne Prolog). Tit.: Incipiunt dicta Leutoldi sublectoris Bienne que intitulantur Speculum de corpore Christi. (163v) Tit.: Sermo in die parasceves. – Inc.: Rex David cooperto capite etc. (?) R. (CAO Nr. 4650). Historia hec est de David quomodo planxit filium suum Absolon. Historia nota est scilicet quid significat … – Expl.: … audiamus illam dulcissimam vocem et vocacionem. Venite benedicti etc. quod ipse prestare dignetur. (165r) De s. Georgio (Nr. 234). Expl. (geringfügig erweitert): … a dracone cum XXti milibus qui conversi sunt ad dominum. – (166v) De s. Philippo et Iacobo (Nr. 238). – (167v) In inventione s. crucis (Nr. 239). Expl.: … et quanta passus sit pro te et omnia erunt tibi dulcia. (168v) Tit.: In diebus rogacionum. – Inc.: Omnis qui petit accipit etc. Ioh. (recte: Mt 7,8 oder Lc 11,10). In verbis istis notandum quod ad impetrandum salutaria 4or oportet considerare Cod. 617 307 v idelicet a quo sit petendum quid sit petendum qualiter sit petendum quando sit petendum … – Expl.: … Expelle eos quoniam irritaverunt te domine. (169r) Tit.: De eodem. – Inc.: Petite et accipietis. Luc. IX (recte: 11,9). In hiis verbis notandum est quod sex sunt que invitant nos ad petendum … – Expl.: … Sextus gradus est quod solem suum facit oriri super bonos et malos. (170r) Tit.: In domini ascensione. – Inc.: Quo abiit dilectus tuus o pulcherrima mulierum. Can. VI (1). In hiis verbis tria videnda sunt scilicet que sit dilecta quis sit dilectus quo et quomodo abiit. Circa primum sciendum quod dilecta est beata virgo … – Expl.: … Vado parare vobis locum ad hunc locum nos perducere dignetur. (171r) Tit.: De eodem. – Inc.: Non est vir in domo sua abiit via longissima secum tulit sacculum pecunie in die plene lune reversurus Prover. VIIIo (recte: 7,19–20). Verba proposita possunt esse interioris hominis id est anime respondentis ad questionem sibi factam … – Expl.: … super malos maledicendo dicens Ite maledicti. (172r) Tit.: De sancto spiritu. – Inc.: De excelso misit ignem in ossibus meis et erudivit me R. (Lam 1,13). Quantum ad presentem festivitatem possunt hec verba esse matris ecclesie … – Expl.: … de iudicio quod non timuerunt ad quam iusticiam nos perducat. (173r) Tit.: De corpore Christi. – Inc.: Accipite et manducate ex hoc omnes (vgl. Mt 26,26; 1 Cor 11,24). Bene fit illi homini qui hunc panem digne et recte manducat … – Expl.: … ad omnia opera bona insuper illo die cicius exauditur. Vgl. Zwettl, Stiftsbibl., Cod. 115, Bl. 12r. (173v) Tit.: De eodem. – Inc.: Rex Assuerus tercio anno imperii sui fecit grande convivium cunc tis principibus et pueris suis et vocavit omnem populum qui inventus est in Susia Hester primo (3). Per hoc convivium Veteris Testamenti figuratum est nobis convivium a Christo hodie institu tum sacri corporis et sanguinis sui … – Expl.: … digne et devote in hospicio cordis sui locaverit benediccionem eternam obtinebit. (174r) De s. Iohanne Baptista (Nr. 245). Expl.: … a puericia crevisse in heremo. Rogemus. (175v) Tit.: De sancto Petro apostolo. – Inc.: Petre amas me. Tu scis domine quia amo te Mt. (recte: Ioh. 21,17). Dicit beatus Iohannes in suo ewangelio quod dominus interrogavit beatum Petrum de sua dileccione dicens … – Expl.: … Ecce quanta passus est beatus Petrus propter amicum suum Christum. (176v) De s. Margareta (Nr. 252). – (177v) De s. Maria Magdalena (Nr. 254). – (178v) De s. Iacobo (Bibelzitat und Expl. wie Nr. 257). Inc.: Dic ut sedeant hii duo filii mei etc. Mt. 20 (21). Filius Dei volens loqui Iudeis dixit hec verba que eciam ego possum dicere. Mea doctrina non est mea sed eius … – (180r) De s. Petro ad vincula (Nr. 149). Expl.: … et regni eius non erit finis (Expl. wie Neustift, Stiftsbibl., Cod. 78, Bl. 72ra). – (181r) In assumptione BMV (Nr. 265). Expl.: … honorificentia populi populi (!) mei scilicet generis humani. (182v) Tit.: De eodem festo. – Inc.: Extendit Noe manum et comprehensam columbam in archam intulit. Gen. (8,9). Hic tria possunt notari scilicet Marie assumpcio cum dicitur Extendit Noe manum 2o festivitas venerabilis Marie cum dicitur Apprehensam columbam 3o assumpcionis securitas cum dicitur In archam … – Expl.: … et scio quia voluntas non abest cum sis mater misericordie obtine nobis graciam filii tui ut a te non separemur. (183r) Tit.: Bartholomei apostoli. – Inc.: Percusserunt me et vulneraverunt me custodes muro rum. Cant. V (7). In verbis propositis notatur triplex martirium quod suscipit beatus Bartholo meus fuit enim fustigatus deinde vivus excoriatus postremum decollatus … – Expl.: … non vult cognoscere celi gaudia et penas inferni. Parallelüberlieferung: München, UB, fol. 64, Bl. 241rb. (184v) De s. Augustino (Nr. 272). Expl.: … sic ipse fugat tenebras infidelitatis. Rogemus etc. – (186r) In nativitate BMV (Nr. 276). 308 Cod. 617 (187Ir) Tit.: In exaltacione crucis. – Inc.: Ego dominus exaltavi lignum humile et humiliavi lig num sublime Ezech. (17,24). In hiis verbis agitur de duobus lignis. Primo de ligno salvacionis in quo homo fuit vivificatus … – Expl.: … ergo ut congregemur. Rogemus dominum etc. (187Iv) Tit.: De sancto Matheo apostolo. – Inc.: Sequere me. Mt. 9 (9). In illo ewangelio duo notantur scilicet dignitas vocantis cum dicitur Sequere me … – Expl.: … quia omnes fideles Christus toto corde semper dilexit. Rogemus etc. (187v) De s. Michaele et de s. angelis (Nr. 286). Expl.: … a nobis sunt honorandi. Rogemus etc. (188v) Tit.: Simonis et Iude. – Inc.: Nimis honorati sunt amici tui Deus Ps. (138,17). Dicitur vulgariter Bonum ferrum trahit homini argentum de bursa … – Expl.: … dicit propheta Nimis honorati sunt amici etc. (189r) Tit.: De omnibus sanctis. – Inc.: Gaudete et exultate omnes sancti quoniam merces vestra multa est in celo Mt. (recte: Lc 6,23). Verba proposita leguntur in ewangelio hodierno et per ea insinuatur nobis gloria premium omnium sanctorum quorum hodie festivitas celebratur … – Expl.: … Ideo debemus hunc diem sermone honorare. Rogemus ergo dominum ut per intercessio nem omnium sanctorum suorum perducat nos ad gaudia sine fine permansura. Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 443, Bl. 131rb. (190r) Tit.: De animabus. – Inc.: Animas pauperum tuorum ne obliviscaris in finem Ps. (74,19). Notandum quod quintuplex est anima scilicet anima inmunda anima rapta anima sepulta anima salvata anima cruciata … – Expl.: … absolvunt animas quas purgans detinet ignis de hiis require in Peregrino de sanctis. (190v) De s. Martino (Bibelzitat und Inc. wie Nr. 174). Expl.: … et reliquam partem induit sequenti nocte vidit celum apertum et Christum etc. ibi dic legendam. (191v) Tit.: De sancta Katherina. – Inc.: Inspice et fac secundum exemplar quod tibi in monte monstratum est (Ex 25,40). Nota quod Deus duo genera sanctorum habet in celo quosdam ab scondit a nobis et quosdam nobis monstravit … – Expl.: … et cum sponso suo in eternum valemus conregnare Amen. (191v) Schlussschrift mit Schreibernennung und Datierung: Explicit per Erasmum Lämpl cano nicum Novecellensem et plebanum in Ässnigk in die Clavorum et Lancee anno Lmo quinto (18. April 1455). 7 (194r–195r) ‹Iohannes Contractus›: Sermo de s. Andrea (Schneyer 3, 439, Nr. 88, zu München, BSB, Clm 3420). Expl.: … deterremur et tamen vocantem sequi contempnimus. Darunter Federproben: 1) Vermerk: Accipe o homo exemplum qui es in periculis huius mundi a sancto Andrea … ad primam vocem omnibus relictis eum sequebatur. 2) Alphabet. 3) Notizen: Venerabilis domine N. Vos misistis ad me pro Amigdalis ego deberem nos providere cum Amigdalis ego locutus sum cum domino N. – Benedictum sit dulce nomen domini mei Ihesu Christi et gloriosissime virginis Marie matris eius in seculum et ultra nos cum prole pia benedicat virgo Maria. Benedictum. (195v) nur Rahmung. 8 (196r) Exzerpt aus ‹Robertus Holcot›: Commentarius in librum Sapientiae, cap. 3, lectio 31, art. 1 (Druck: u. a. GW 12884. – Stegmüller RB Nr. 7416. Kaeppeli Nr. 3497). [A]brahe sinus dicitur requies sanctorum qui olim fuit limbus nunc est celum et sic orat ecclesia sinus te suscipiat. (196v–199v) nur Rahmung. 309 Cod. 654 IV (200[201]–281[282]) B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen in T. I. Lagen: 3.VI235(236) + V245(246) + 3.VI281(282). Lagenreklamanten. In den Lagenmitten beschrie bene Pergamentfälze. Zw. vorletzter und letzter Lage Reste eines Papierfalzes, wohl zum urspr. als Spiegel auf dem HD aufgeklebten, herausgerissenen Blatt gehörig. S: Schriftraum 160 × 105, von feinen, tw. kaum sichtbaren Tintenlinien umrahmt, zu 34 Zeilen. Bastarda. A: Platz für zweizeilige Lombarden freigelassen. Lediglich Bl. 205v–207v, 218v–220r, 227v–229r, 250v–253r und 277v–280r rubriziert: Auszeichnungsstriche, Unterstreichungen, Notizen, Abschnittszählungen. Bl. 201v Zeigehand. Satzmajuskeln und einzelne Buchstaben der ersten Zeilen tw. gelängt. 9 (200r–280r) ‹Servasanctus de Faenza›: Sermones (Schneyer 5, 388–391). (200v) De apostolis (Nr. 193). Expl.: … et que sunt infra nos mundum. Rogemus igitur omni potentem. (202v) Nr. 194. (205v) Nr. 195. (207v) Nr. 196. – (210v) De evangelistis (Nr. 198). (212v) Nr. 199. – (214r) De uno martyre (Nr. 200). (217r) Nr. 201. (218v) Nr. 202. (220r) Nr. 203. (222r) Nr. 204. – (223v) De pluribus martyribus (Nr. 205). (226r) Nr. 206. (227v) Nr. 207. (229r) Nr. 208. (231r) Nr. 209. – (233v) De uno confessore (Nr. 210). (236v) Nr. 211. (239r) Nr. 212. (241r) Nr. 213. (243v) Nr. 214. – (246r) De pluribus confessoribus (Nr. 215). (248v) Nr. 216. – (250v) De uno confessore (Nr. 217). (253r) Nr. 218. – (256v) De una virgine (Nr. 219). (258r) Nr. 220. (260r) Nr. 221. (261v) Nr. 222. (263v) Nr. 223. Expl.: … quibus carui dum ista dictarem Valete fratres et orate pro me dominum nostrum Ihesum Christum. (267v) Nr. 188 bzw. Schneyer 7, 643, Nr. 9, zu Köln, Stadtarch. GB, fol. 146. Expl.: … Sic paupertatem amare sic obedire etc. (= Expl. Schneyer 7, 643, Nr. 9). (269v) Nr. 191. (271v) Nr. 197. Expl.: … confundit animam. – (274v) De BMV (Nr. 224). (277v) Nr. 225. (279v) Nr. 226. Expl.: … scias te esse vacuum et non plenum etc. Amen. Deo gracias. (280v–281r) nur Rahmung. (281v) Besitzvermerke (s. bei G). U. S. Cod. 654 LIBER HORARUM, Bd. 2 (zusammengehörend mit Cod. 353) Perg. II, 207, I Bl. 165 × 120. Raum Brixen, 1496. B: Regelmäßiges Perg., sehr vereinzelt kleine Löcher und Nahtspuren. Lagen: (I–1)I (Vorsatzbl., Pap. 20. Jh.) + (I–1)II + VI10a(12) + 8.V90(92) + (V–1)99(101) + (V–1+1)109(111) + 9.V197(201) + (IV–2)203(207) + (I–1)I* (Nachsatzbl., Pap. 20. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. II, 91, 100, 199 und 200 herausgeschnitten (ohne Textverlust). Bl. 109 eingehängtes Einzelbl. Am oberen Blattrand zeitgenössische rote Foliierung in arabischen Ziffern von 1 bis 202, anschließend ein ungezähltes Bl. (ursprünglich wohl als Spiegel am HD aufgeklebt), Fehler in der Zählung: 7 und 119 doppelt angeführt, zw. Bl. 10 und 11 sowie zw. Bl. 191 und 192 jeweils ein ungezähltes Blatt. S: Schriftraum 110 × 70, von zweifachen roten Tintenlinien gerahmt, zu 17–19, größtenteils einheitlich 19 Zeilen auf feiner Bleistift-, tw. Blindlinierung. Zirkelstiche. Kalligraphische Bastarda (auffällig gelängte Initialmajuskeln, Unterlängen und Kürzungsstriche, bisweilen i-Punkte in Kreisform), dat. 28. Juli 1496 (Bl. 202r). Schreiber: Georg Hölzl aus Mattighofen, vgl. Krämer, Scriptores. A: Rote Auszeichnungsstriche, (Zwischen-)Überschriften, Interpunktionszeichen, Unterstreichungen, Zeilen füllsel (Bl. 133r, 144v); wenige rote und blaue Paragraphzeichen. Bis Bl. 119r ein- bis sechszeilige Lombar den, abwechselnd hell-/dunkelrot und blau, mit Punktverdickungen bzw. gestielten Punkten; Bl. 16v Lom barde nicht ausgeführt. Repräsentanten. Deckfarbenschmuck: 18 vier- bis siebenzeilige ornamentale Initialen mit Ranken; 26 ganzseitige (Schrift spiegelgröße) rankenlose Miniaturen, die ab Bl. 122r in drei gleiche, querrechteckige Felder untergliedert 310 Cod. 654 sind (Bl. 144r oberes und unteres Register zusätzlich in zwei quadratische Felder unterteilt), sodass sich, einschließlich des Kruzifixes Bl. 93v, insgesamt 60 Bilder ergeben. Die Ornamentalinitialen (Bl. 1r, 16r, 34r, 52r, 67r, 91r, 94r, 100r, 110r, 117r, 123r, 133r, 144v, 155r, 165v, 179r, 188v, 198v) mit meist annähernd quadratischen, durch Konturlinien gerahmten Initialfeldern; Buchsta benkörper in der Regel mit großformigen Blattranken gefüllt; Bl. 117r etwas filigranere Initiale mit Ausläu fer (Achterschlinge); folgende Binnenfeldgestaltungen in Deckfarbe: geometrische Muster, gegenständige Blattpaare, typisch das aus dem Federzeichnungsdekor der Zeit übernommene Motiv der aneinanderge reihten Kopfstempel, die als Negativformen blattartige Gebilde entstehen lassen (z. B. Bl. 188v; vgl. auch Brixen, Priesterseminarbibl., Cod. C.8); Bl. 165v Binnenfeldblätter mit „Erbsenschotenmotiv“ und Bl. 91r, 100r, 110r, 155r Perlreihen, die von den Füllblättern des Buchstabenkörpers umschlossen werden. – Die Akanthusranken (mit variierenden Blattformen, u. a. mit tropfenförmig verdickten Adern) ohne Kontakt zur Initiale, meist um die untere rechte Blattecke gelegt, dabei in der Ecke eine goldgefüllte Schlinge ausbil dend; als Dekor Goldpunkte, auch frei (mit meist rotem Federzeichnungsdekor), bisweilen Fadenranken; als Randschmuck auf einigen Folios naturalistische Zweige, die recht grob gemalt sind: Bl. 52r Distel, 91r Glockenblumen (?), 94r Akelei (?), 110r Erdbeeren, 117r Heckenrosen; Bl. 34r Akanthusranke mit naturali stischen Anschnitten. – Zur Gestaltung von Initialen (Buchstabenkörper und Binnenfelder) und Ranken s. auch Cod. 353. Die Rahmen der Miniaturen in Gold oder Deckfarbe (einfarbig sowie verschiedenfarbige Abschnitte im Wechsel) mit Kontur- und Gehrungslinien; Bl. 15v sowie Cod. 353, Bl. 93v, zusätzlich mittig an den Rah menleisten Goldpunkte mit roter Federzeichnungszier (Fadenbündel mit Fibrillen); diese, in üppiger Ausfor mung, auch am Kruzifix Bl. 93v. In die ganzseitigen Miniaturen sind tw. Architekturrahmen eingeschrieben (Rundbogen, fallweise mit Zwickelblättern, Knospenkapitellen und -basen); in Cod. 654 werden auch durch die architektonischen Gegebenheiten (Bogen, Säulen, Durchgänge) Rahmungen geschaffen, die die Einzel felder untergliedern. Hintergründe: nur selten Teppichgründe (rosafarben mit groben, karminroten Ranken; Bl. 11v und 144r unten rechts) und Goldgrund (v. a. für die Marienfiguren, s. Bl. 109v sowie Cod. 353, Bl. 130v; zu Punzierungen s. u. und Cod. 353); häufig nackte Mauerflächen (rosafarben oder grau); gerne auch Landschaftshintergründe (Fensterausblicke), diese simpel und mit wenigen Details angelegt, zu denen kleine Boote auf den Seen bzw. Flüssen gehören; etwas aufwändiger allerdings Bl. 15v, 177v (Mitte), 178r (oben) sowie Cod. 353, Bl. 54v, 70v; Wolken als ondulierende Bänder dargestellt; charakteristisch die Vogel formationen in helleren Himmelszonen; Vegetation der grünen Hügel nur durch horizontale Strichelung angedeutet, auch die meist breitblättrigen Pflanzen im Vordergrund eher grob, nur wenige Blumen differen zierter dargestellt (z. B. Bl. 177v oben und Cod. 353, Bl. 130v: Maiglöckchen). – Figuren mit uniformen Gesichtern (lange, zuweilen etwas klobige Nasen, kleine, rundliche Münder mit leicht nach unten gezogenen Mundwinkeln und rundbogige Augenbrauen), rosigem Inkarnat und einfachen, aber lebhaften Haarstruk turen (Marienfiguren mit langem, gewelltem Blondhaar); größere Heiligenscheine (v. a. von Gottvater und Christus) als Kreuznimben, das Kreuz in Rot eingetragen, tw. ergänzt durch Strahlenkranz (helleres Mal mittel); Körper von Draperien verhüllt, deren Faltenwürfe durch lapidare Schattierungen, Schraffen und Höhungen herausgearbeitet sind, z. B. Schutzmantel-Madonna, Bl. 116v; Maria Magdalena stets mit zwei farbiger, turbanartiger Kopfbedeckung; ungelenke Anatomie des Gekreuzigten (Gliedmaßen im Verhältnis zum Rumpf zu kurz, tordierte Beine), der im Vergleich zum Kreuz meist zu klein angelegt ist, z. B. Bl. 93v; sein Lendentuch häufig abflatternd dargestellt. – Perspektivische Unsicherheiten, z. B. Bl. 144r (Mitte) die Fluchtlinien der Seitenwände; auch Einrichtungsgegenstände perspektivisch uneindeutig in das Raumgefü ge eingezeichnet, z. B. die Pulte, auf denen jeweils ein aufgeschlagenes Buch liegt. Buntfarbiges Kolorit u. a. mit klarem (Kobalt-)Blau und charakteristischem Gelbgrün, u. a. für Holzflächen; gerade von Gesichtern Deckfarbenschicht immer wieder abgeplatzt, z. B. Jesus, Bl. 153v (Mitte) oder auch Brust des rechten Schergen und Gesicht Jesu, Bl. 164v (oben); Blattgold nur wenig abgerieben, z. B. Binnenfeld der Ornamen talinitiale Bl. 67r, hier lachsroter Bolus sichtbar. Bl. 11Iv Allerheiligen: Heiligenschar; in der vordersten Reihe (von links nach rechts): Katharina (Schwert und Bruchstück eines Rades), Andreas (X-Kreuz), Petrus (Tiara und Schlüssel), Sebastian (Pfeil) und Bene dikt (Stab und Nuppenglas). Bl. 15v Auftraggeber-Bildnis: Junger, blondgelockter Mann in schwarzem, pelzbesetztem Mantel, wohl Kaspar Neuhauser, kniet mit gefalteten Händen und mit Gebetsschnur vor Christus an einem hohen Kreuz, das in einer Landschaft (von kleinen Städten übersäte Ebene und See oder Fluss vor einer Bergkette) auf gestellt ist; Gekreuzigter mit gelbgrünem Lendentuch. Am oberen Rand des Himmels Textabklatsch von Bl. 16r; über den Schwalben gelbe stilisierte Wolken (?). Bl. 33v Armenseelenbild: Auf einem Friedhof kniet betend ein junger, blondgelockter Mann (mit s chwarzem Umhang und roten Hosen) vor dem Beinhaus, während aus mehreren Gräbern mit Dreschflegel, Pfeil und Bogen, Schere etc. ausgestattete Skelette entsteigen, sich ihrer Leichentücher entledigen und bewaffnete Angreifer verjagen; neben dem Beinhaus, an dem außen ein Weihwasserkessel (mit Aspergil) angebracht ist, Kirche mit grünem Dach, Maßwerkfenstern und -rosette sowie Portal mit beschlagwerkartigen Verzierungen. – Cod. 654 311 Dargestellt ist die Rettung eines von Feinden bedrohten Beters durch die Armenseelen, die ihm aus Dank barkeit für sein Gebet zu Hilfe eilen (s. Wieser). Bl. 51v In einem Kirchenraum mit blauem, von weißen Kreisen übersätem Rippengewölbe und Maßwerk fenstern sitzen drei Kleriker, vor denen (mit dem Rücken zu ihnen) ein Büßer, möglicherweise Kaspar Neuhauser, mit entblößtem Oberkörper auf dem nackten Steinfußboden kniet und eine brennende Kerze vor sich hält. Bl. 66v (vgl. Bl. 33v) Auf einem Friedhof beten ein junger, blondgelockter Mann (in langem, schwarzem Gewand) und eine Frau (mit Rise und schwarzem Kleid), vermutlich Kaspar Neuhauser und seine Gemahlin, die eine Tochter von Michael Pacher war (s. C. Fischnaler, Die Erben Michael Pachers. Zeitschrift des Fer dinandeums für Tirol und Vorarlberg 37 [1893] 359–361). Der Mann kniet vor dem Beinhaus, die Frau im Vordergrund, tw. von der Kirchhofmauer verdeckt; zw. ihnen Gräber und Gebeine; beide Eheleute mit Ge betsschnüren; neben dem Beinhaus eine Kirche (ähnlich Bl. 33v, aber größer); sowohl an der Kirche als auch am Beinhaus Weihwassergefäß angebracht, auch neben der Frau ein Weihwasserkessel, jeweils mit Aspergil. Bl. 93v Goldenes Kreuz mit den Evangelistensymbolen (jeweils mit Spruchband) in Medaillons (mit Drei passbesatz) an den Kreuzenden (oben beginnend, im Uhrzeigersinn): Johannesadler, Markuslöwe, Lukas stier, Matthäusengel; große Goldpunkte in den Winkeln der Kreuzarme (zum Federzeichnungsdekor s. o.). Der Gekreuzigte mit weißem Lendentuch. Bl. 109v Maria im Ährenkleid. Bl. 116v Schutzmantel-Madonna als Pestbild: Maria in goldenem Gewand versammelt unter ihrem blauen Mantel Frauen und Männer aller Stände (Papst und König deutlich erkennbar), die kniend beten. Im Himmel Gottvater mit Pfeil und Bogen als Symbole seines Zornes, der als Pest über die Menschen käme, wenn Maria sie nicht davor schützte. Bl. 122r oben Zwei Engel mit blau-roten Flügeln verkünden Anna und Joachim die Geburt Mariens. – Mitte: Joachim umarmt Anna. – unten Geburt Mariens: Anna liegt mit Maria (als Wickelkind) im Bett und b ekommt von einer Magd einen Teller mit Essen gereicht, das eine Köchin in der angrenzenden Küche zubereitet hat. Bl. 122v oben der Tempelgang Mariens: Maria als Mädchen im Ährenkleid steigt betend eine Treppe empor, an deren Fuß ihre Eltern und oben ein Geistlicher (mit Bischofshut) stehen. – Mitte: Maria im Tempel (links sieht man sie diskutierend zw. zwei Jungfrauen, rechts lesend an einem Pult). – unten Vermählung Mariens: Bischof hält Josef und Maria, die ihn flankieren, jeweils an den Unterarmen; zwei Zeugen. Bl. 132r oben Verkündigung (Engel, Schriftband AVE GRACIA PLENA um Säule geschlungen, Maria be tend an einem Pult; über ihr die Taube, im Hintergrund grüner Vorhang). – Mitte Heimsuchung: Vor einem Haus mit rundbogigem Tor begrüßt Elisabeth Maria. – unten Geburt Christi: Im Stall, hinter dem Ochs und Esel hervorlugen, beten Maria und Josef (mit Kerze) kniend das auf dem Boden liegende, nackte Kind an. Bl. 132v oben Beschneidung Christi: Auf einem Holzthron ein greiser Priester; auf seinem Schoß das Jesus kind, das vom knienden Mohel kniend beschnitten wird; zwei Assistenzfiguren. – Mitte Anbetung der Hei ligen Drei Könige: Vor dem Stall sitzt Maria mit dem nackten Kind auf dem Schoß; ihre Schattensilhouette zeichnet sich im Stallinneren ab; der dunkelhäutige König hebt seine Krone zum Gruß. – unten Darbringung Christi im Tempel: Neben dem Altar (mit gemustertem Tuch, darauf die Gesetzestafeln) links Frau und Priester mit dem Jesuskind, rechts Maria (mit gefalteten Händen) und Josef mit den Opfertauben. Bl. 143v oben links schläft Josef an einem Wangentisch; im Traum erscheint ihm ein Engel, der ihm befiehlt, mit seiner Familie zu fliehen; rechts stillt Maria das Kind. – Mitte: Flucht nach Ägypten. – unten Christus unter den Schriftgelehrten: links doziert Jesus von einem erhöhten Sitz herab, rechts steht er bei seinen Eltern. Bl. 144r oben links Maria spinnend; rechts Josef als Zimmermann bei der Arbeit (Vermessen?), bei der ihm Jesus hilft (auf dem Boden Beil und Farbtopf?); deutliche Risse in der Mauer im Hintergrund. – Mitte: Josef, Jesus (als Jüngling) und Maria bei Tisch. – unten: links kniet Jesus betend vor einem Pult, rechts sitzt Maria lesend an einem Pult. Bl. 153v oben Taufe Jesu im Jordan: am Ufer ein Engel mit den Gewändern Jesu. – Mitte Versuchung Jesu: flankiert von kahlen Bäumen Jesus und der Teufel (mit roten Vogelbeinen und hängenden Brüsten). – unten Berufung der Apostel Petrus und Andreas: links Petrus und Andreas als Fischer, die auf dem See von Galiläa ihre Netze ausbringen; rechts, am Seeufer, Jesus und die beiden Apostel im Boot. Bl. 154r oben Hochzeit zu Kana: links Jesus, der das Wasser (in sechs hohen Krügen, dahinter Maria) in Wein verwandelt; rechts die Hochzeitsgesellschaft. – Mitte: Jesus predigt von einer Holzkanzel herab zu einer sitzenden Gruppe, dahinter stehen Maria und zwei Apostel. – unten: Heilung des Lahmen. Bl. 154v oben Auferweckung des Lazarus: links Martha (rothaarig) und Maria (hält ihr Gewand vors Ge sicht), die Schwestern des Lazarus; der Auferweckte steht, umhüllt von einem rosafarbenen Leichentuch, betend im Grab. – Mitte Einzug nach Jerusalem: Jesus reitet segnend auf der Eselin dem Stadttor zu, vor dem ein Mann einen roten Mantel ausbreitet. – unten Verrat des Judas: links ein Bischof auf einem verzierten Holzthron, der mit einem Mann mit roter Haube diskutiert; rechts ein Hohepriester und der gelbgewandete, rothaarige Judas mit rotem Geldbeutel. 312 Cod. 654 Bl. 163v oben Letztes Abendmahl an einem runden Tisch; Johannes kauert auf dem Schoß Jesu, Judas (wie Bl. 154v) deutlich erkennbar. – Mitte Ölberg: Jesus betet Blut schwitzend den Kelch im Felsen an, während die Jünger schlafen; Felsen als sandfarbene, kulissenartige Formationen mit gewellten Konturen gestaltet. – unten Gefangennahme Jesu (Judaskuss). Bl. 164r oben: Drei Apostel mit Maria Magdalena und Maria. – Mitte Verhöhnung: Pilatus als Bischof; Jesus mit Blutschweiß und verbundenen Augen. – unten Geißelung: auf dem Bein des rechten Schergen ein goldenes M mit Krone darüber. Bl. 164v oben Dornenkrönung; der grüne Rock des linken Schergen mit blattförmigen Schößen. – Mitte: links wäscht Pilatus seine Hände, rechts wird Jesus vorgeführt (einer der Soldaten mit geöffnetem Mund). – unten Kreuztragung: links Maria und Johannes; Simon von Cyrene hilft Jesus beim Tragen. Bl. 165r oben Kreuzigung: links Johannes stützt Maria; Gekreuzigter mit gelbgrünem Lendentuch. – Mitte Pietà, flankiert von Johannes und Maria Magdalena (mit Salbgefäß), im Hintergrund Querbalken des Kreu zes. – unten Grablegung Christi: Josef von Arimathäa bettet Jesus in Steinsarkophag, vor dem Johannes (als Rückenfigur), Maria (mit erhobenen, gefalteten Händen) und Nikodemus knien; stehend Maria Magdalena. Bl. 177v oben Höllenfahrt Christi: Jesus (mit Siegesfahne) befreit die Seelen der Gerechten, die ihm aus dem zerborstenen Höllentor entgegeneilen; aus Seitenfenster der Pforte blickt ein grüner Teufel. – Mitte Auferstehung Christi: Der Auferstandene in rotem Mantel (mit Wundmalen und Siegesfahne) steht vor dem geschlossenen Sarkophag; die schlafenden Soldaten verdeckt; Seelandschaft, darin u. a. Golgota, Baum mit Erhängtem, drei Figürchen, im Hintergrund Bergkette. – unten: Christus erscheint Maria, die betend an einem Pult sitzt. Bl. 178r oben Noli me tangere: In einem von einem Flechthag (vgl. Bl. 163v) umzäunten Garten kniet Maria Magdalena (mit Salbgefäß) und betet Christus an, der segnend und mit Siegesfahne vor ihr steht; in der Hintergrundslandschaft u. a. Golgota sowie drei Soldaten vor einem Wirtshaus (?). – Mitte: Christus (segnend und mit Weltkugel) erscheint zw. vier Frauen (darunter Gottesmutter erkennbar) und drei Aposteln (zuvorderst Petrus, mit Schlüssel), denen er sich zuwendet. – unten Himmelfahrt Christi: Maria und Apostel flankieren den Auffahrtsberg (darauf Fußspuren Christi); aus dem Wolkenband ragen noch Füße und Ge wandsaum Jesu sowie das untere Ende der Siegesfahne (?) heraus. Bl. 178v oben Pfingsten: Taube des hl. Geistes über Maria und Aposteln. – Mitte: Petrus predigt von einer verzierten Holzkanzel (dahinter weitere Apostel) zu einer sitzenden Gruppe herab, hinter der Maria und zwei Frauen stehen. – unten Letztes Gebet Mariens: Maria sitzt betend an einem Pult, links ihr Sterbebett (dahin ter Vorhang), rechts (an der Tür) drei Apostel. Bl. 187v oben: Engel (mit grünblättrigem Zweig) verkündet Maria, die an einem Pult sitzt, den Tod. – Mitte Marientod I: Die verstorbene Maria liegt mit geschlossenen Augen auf dem Bett; drei Apostel. – unten Marientod II: Umgeben von sechs Aposteln liegt die Verstorbene auf dem Bett, über ihr in einer G oldgloriole (mit Wolkenrand) Christus, der das Seelenfigürchen Mariens auf dem Arm hält. Bl. 188r oben Himmelfahrt Mariens: Maria wird von sechs Engeln, zwei davon musizierend (Harfe und Laute), in den Himmel empor getragen. – Mitte: Maria, deren blaues Kleid von Goldglanz überzogen ist, flankiert von zwei weiblichen und vier männlichen Heiligen (davon zwei mit Schriftbändern: IMAR[…]O und HVEM). – unten Krönung Mariens: Maria kniet vor der Dreifaltigkeit; links Christus und rechts Gott vater, die ihr die Krone aufsetzen, hinter ihr der hl. Geist in Menschengestalt. Punzierungen. Rautengitter aus winzigen Quadraten, im Binnenfeld Bl. 67r mit fünfblättrigen Blüten (aus kleinen Kreisen) gefüllt, Bl. 94r, auf kobaltblauem Deckfarbengrund, mit zwölfblättrigen Blüten und auf dem Miniaturenhintergrund Bl. 109v mit sechsblättrigen, abwechselnd rot und blau ausgemalten Blüten, Gitterschnittpunkte zudem mit vierblättrigen roten und blauen Blüten besetzt. Deckfarbenschmuck von der Hand, die auch Cod. 353 illuminierte. Während Cod. 353 einige Detailfreude zeigt (ganzseitige Miniaturen, die mehr Gestaltungsfläche bieten), überwiegen in Cod. 654, bis auf Bl. 11v (Heiligenfiguren etwas individueller gestaltet), naiv-schematisierte Illuminationen; in beiden Bänden far benfreudiges Kolorit und verhältnismäßig reicher Einsatz von Blattgold sowie Punzierungen. – Der Illumi nator von Cod. 353 und 654 kannte sehr wahrscheinlich das wohl zu Beginn der 1480er Jahre in Neustift entstandene Epistolar (Cod. 196, Bl. 1r–20v) und Evangelistar (Esztergom, Kathedralbibl., Ms. II.2, und Cod. 196, Bl. 30r–35[26]v), s. die Blattgoldgründe mit gepunztem Rautengitter und sowohl punzierten als auch aufgemalten, bunten Blüten (vgl. Cod. 353, Bl. 1r, mit Cod. 196, Bl. 10v1), den um eine Folioecke geführten Randschmuck, der eine goldgefüllte Schlinge ausbildet (z. B. Ms. II.2, Bl. 3r und 11r; gerade im Evangelistar beginnt sich der Randschmuck vollständig von der Initiale zu lösen, wie das in Cod. 353 und 654 durchgängig der Fall ist; s. z. B. Ms. II.2, Bl. 34v, wo die Randzier die Initiale nur noch hauchfein berührt), die naturalistischen Zweige (vgl. z. B. Cod. 353, Bl. 94r mit Cod. 196, Bl. 34r: roter Klee) und generell den Akanthus mit seinen von Seite zu Seite variierenden Blattformen. – Als Vorlagen für die Minia turen Cod. 654, Bl. 116v–188r, dürfte der Buchmaler die Holzschnitte aus dem Inkunabeldruck des Itinera rium seu peregrinatio BMV (s. bei Inhalt, Nr. 9) herangezogen haben, von denen er auch die dreiregistrige Unterteilung der Bildfelder übernahm. In Frage kommen sowohl die ca. 1487/88 von Johann Reger in Ulm Cod. 654 313 gedruckte lat. Version dieses Werks (GW M15415) als auch die 1489 von Johann Schobser in Augsburg ge druckte dt. Fassung (GW M15419); die Holzschnittzyklen dieser beiden Drucke sind nicht völlig identisch, aber stark verwandt, und zeigen im Vergleich mit den Miniaturen in Cod. 654 mehr Übereinstimmungen als (geringfügige) Abweichungen. Bei dem M mit Krone (Cod. 654, Bl. 164r unten) könnte es sich um eine Buchmaler-Signatur handeln, wobei sich die Frage stellt, warum gerade einer der geißelnden Schergen dieses Zeichen trägt. Cod. 353 ist unsigniert. S. R. E: Renaissance-Einband: braunes Leder auf Holz mit Golddruck. Neustift, zw. 1569 und 1581 (identisch mit Cod. 353). Werkstatt: Narziß Schuechl. Einband restauriert, Leder tw. ergänzt (deutlich sichtbar), Bindung erneuert. VD und HD gleich: ein von je zweifachen Streicheisenlinien in Gold begrenzter Rahmen: Rolle Enthauptung des Johannes, Eherne Schlange, Kreuzigung, Auferstehung (Nr. 1 = Neuhauser, Rolleneinbände, R 29). Im Mittelfeld des VD Plattenstempel mit Darstellung eines Erzengels (Michael?) mit Wappenschild von Propst Augustin Schabl (Propst 1569–1581) mit zwei steigenden geflügelten Löwen (Nr. 2 = Neuhauser, Rolleneinbände, P 3). Da rüber Inschrift Augustinus praepositus Novæcelle. Im Mittelfeld des HD in den Ecken Einzelstempel mit floralem Dekor aus Vase wachsend (Nr. 3), in der Mitte Einzelstempel Kreuzigung Christi (Nr. 4). Zwei Leder-Metall-Schließen, beim Materialwechsel sich stark verjüngend, Leder erneuert. Kanten gerade und nach innen hin abgeschrägt und durch einfache Streicheisenlinie eingefasst. Rücken: vier Doppelbünde, von zweifachen goldenen Linien gesäumt, Linien am VD und HD fortgesetzt, spitz aufeinander zulaufend. In den Feldern zw. den Bünden jeweils florale Rolle (Nr. 5 = Neuhauser, Rolleneinbände, R 26). Kapitale mit naturfarbenem und rotem Zwirn umstochen (20. Jh.). Schnitt goldfarben und punziert. Spiegel Pap. 20. Jh. Am Spiegel des VD Signatur (s. bei G). G: Hs. im Auftrag des Kaspar Neuhauser, bischöflicher Zöllner (1490/92) und Stadtrichter (1494–1501) in Klausen (vgl. Kustatscher I 300–302 und II 646; K. Brandstätter, Städtische Verwaltung und Bürgerschaft in Klausen im 15. Jh. Der Schlern 73 [1999] 442, 718), 1496 von Georg Hölzl geschrieben. Hs. spätestens seit der Zeit des Propstes Augustin Schabl (1569–1581) in Neustift. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. IIr sowie 202v Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. IIr Stempel der Soprintendenza Tridentina 20. Jh. Am Spiegel des VD Signatur 20. Jh. Ms 654 (Bleistift). L: Wilhelm II 108 und 109. – Hermann 209–211, Nr. 209. – H. Wieser, Die dankbaren Toten. Veröffentlichungen des Tiroler Landesmuseums Ferdinandeum 26/29 (1946/49) 491–501, Taf. XLVIII. – Laußermayer 6, 301– 308. – Colophons Nr. 5053. – Neuhauser, Neustift 85. – Peintner, Buchmalerei, 33–34, 65–83, 87, 91–92 (ohne Nennung der Sign.). – H. Rizzolli, Der Fachbogen der Hut- und Tuchmacher als spätmittelalterliches Heiligenattribut in Tirol. Der Schlern 60 (1986) 52, 56 (Abb. 4). – Stefani 116. – Peintner, Stiftsbibliothek 121. – Kustatscher I 302 (ohne Nennung der Sign.). – Katalog ULB Tirol VII 299, 485. – Kustatscher– Korenjak 157. – Krämer, Scriptores, s. v. „Höltzl (Hölzl), Georgius“. Liber Horarum, Bd. 2. (Ir–v) leer. (IIr) Tit. von späterer Hand ergänzt (Abschrift von Cod. 353, Bl. Ir): Subscripti cursus sparsim hincinde in devocionum libellis inventi summa cum diligencia hic in unum sunt collecti ut quivis ad excitandam devocionem prout suus afficitur animus ad manum habeat quo se orando convertat: aut scilicet ad tri nitatem beatissimam aut ad eternam sapienciam ad corpus Christi ad ipsius passionem vel eius a nimam ad spiritum sanctum vel beatissimam virginem Mariam ad beatos angelos vel omnes Dei sanctos aut pro misericordia inpetrandam (!) pro peccatis propriis vel fidelibus defunctis cum septem peniten c[ialibus] psalmis cum mortuorum vigiliis et accessu altaris prout secundum ordinem huiusmodi cum numero foliorum hic sunt intitulati. (IIv) Inhaltsangabe mit Blattangabe. Cursus de omnibus sanctis 1 / Cursus pro peccatis 16 / Cursus de animabus 34 / Septem psalmi 52 / Vi gilie mortuorum 67 / Vespere mortuorum 87. 1 (1r–15r) Cursus de omnibus sanctis. Lektionen (‹Ps.-Augustinus›: Meditationes, cap. 24 [PL 40, 918–919. – CPL Nr. 386]). 314 Cod. 654 Die Beschriftung folgt nicht der Blattfolge, der Text auf Bl. 7r wird auf 8v fortgesetzt, an schließend folgt Bl. 7Ir, dann 7v, 8r und schließlich 7Iv, ehe der Text (ohne Textverlust) auf Bl. 9r weitergeht. Der Text von Bl. 10v wird auf 11r fortgesetzt, das Blatt dazwischen (11I) wurde erst später eingehängt. Tit.: Cursus de omnibus sanctis. – Anrufung: Domine labia mea aperies. Et os. Deus in adiuto rium meum. (1r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus (AH 51, Nr. 129). – (1v) Ps 31,20– 25. – (2r) Antiphona. Te mirabilem in sanctis rex regum collaudamus tua opera Criste tua mirabilia … – (2v) Benediccio. Per suffragia beate virginis Marie et omnium sanctorum tu orum … – Leccio prima. Inc.: Felices sancti Dei omnes qui pertransistis pelagus mortalitatis et pervenire … – Expl.: … agitamur procellis. Tu autem. – (3r) Responsorium mit Versikel. – (3v) Leccio secunda. Inc.: Vos porte pulcerrime que in magnam surrexistis altitudinem … – Expl.: … salvi esse non possumus. – (4r) Responsorium mit Versikel. – Leccio tercia. Inc.: O vos electi sumus nanque (!) fragiles et nullius veritatis … – Expl.: … quietis continue pacis et nunquam sinende securitatis. – (4v) Responsorium mit Versikel. – (5r) Te Deum. (5r) Laudes. Ps 150. – (5v) Antiphona. Omnes sancti Dei intercedere pro nobis ad dominum Ihe sum Christum alleluia. – Capitulum. – Hymnus (AH 51, Nr. 130). – (6v) Canticum Zachariae. – (7r) Antiphon (CAO Nr. 4689). – Oracio. Concede quesumus omnipotens Deus ut intercessio sancte Dei genitricis Marie sanctique Iohannis Baptiste … (8v) Prim. Hymnus (AH 51, Nr. 130). – (7Ir) Ps 117. – Antiphon (Cantus Nr. 203647). – Capitu lum. – Responsorium mit Versikel. – (7Iv) Oracio. Deus qui nos concedis omnium sanctorum tuorum merita venerari … (7Iv) Terz. Hymnus (AH 51, Nr. 130). – (8r) Ps 33,1–6. – (8v) Antiphon (CAO Nr. 4583). – Ca pitulum. – Responsorium mit Versikel. – Oracio. Deus qui nos beate Marie semper virginis et beatorum spirituum patriarcharum … (9r) Sext. Hymnus (nur Verweis: AH 51, Nr. 130). – Ps 34,9–15. – (9v) Antiphon (CAO Nr. 3208). – Capitulum. – Responsorium mit Versikel. – (10r) Oracio. A cunctis iniquitatibus nostris exue nos domine et intercedente beata et gloriosa Dei genitrice virgine Maria … (10r) Non. Hymnus (nur Verweis: AH 51, Nr. 130). – Ps 34,16–23. – (10v) Antiphona. O quam digna et veneranda [ (10Ir) Gebet: Corda nostra quesumus domine passionum carismatum et glorie racionalis … (10Iv) ganzseitige Miniatur (s. bei A). (11r) Forts. von Bl. 10v. Inc. mut.: ] est memoria omnium sanctorum Dei qui despecto pharao nis … – Capitulum. – Responsorium mit Versikel. – (11v) Oracio. Infirmitatem paupertatem tribulacionem atque omnem angustiam nostram … (11v) Vesper. Ps 145,10–16. – (12r) Antiphon (CAO Nr. 4063). – Capitulum. – (12v) Respon sorium mit Versikel. – Hymnus (AH 51, Nr. 129). (13r) Magnificat (nur Verweis). – Antiphon (CAO Nr. 3590). – (13v) Oracio. Concede quesumus omnipotens Deus ut intercessio sancte Dei genitricis Marie sanctique Iohannis Baptiste … (14r) Komplet. Ps 145,17–21. – Antiphon (CAO Nr. 2908). – Capitulum. – (14v) Hymnus (AH 51, Nr. 130). – Canticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 3210). – (15r) Ora cio etc. Ut tuam domine misericordiam consequamur fac nos per intercessionem … (15v) ganzseitige Miniatur (s. bei A). 2 (16r–33r) Cursus pro peccatis. Tit.: Cursus pro peccatis multum devotus. Anrufung: Ave benigne Ihesu fili David miserere nobis. Domine labia mea aperies. Et os. Deus in adiutorium. (16r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus (AH 51, Nr. 54, Str. 1–2). – Ps 5. – (17r) Antiphon (CAO Nr. 1847). – (17v) Benediccio. Per magnam misericordiam domini nostri Ihesu Cristi et per multitudinem miseracionum suarum salvet et protegat … – Leccio prima. Inc.: Cod. 654 315 Deus meus confundar et erubesco levare faciem meam … – Expl.: … quoniam Deus miseracio num et clemens tu es. Tu autem (vgl. 2 Esr 9,31). – (18r) Responsorium mit Versikel. – (18v) Leccio secunda. Inc.: Iniquitates nostre contenderunt contra nos domine libera nos … – Expl.: … ne derelinquas nos domine Deus noster. – Responsorium mit Versikel. – (19r) Leccio tercia etc. (Ier 14,19–21). – (19v) Responsorium mit Versikel. – Te Deum (nur Verweis). . (19v) Laudes. Ps 51,3–5. – (20r) Antiphon (vgl. CAO Nr. 4678). – Capitulum. – Hymnus (AH 51, Nr. 54, Str. 3–5). – (20v) Canticum Zachariae (nur Verweis). – Antiphon (CAO Nr. 2770). – Ora cio. Omnipotens et sempiterne Deus cuius natura bonitas cum (!) voluntas potencia cuius opus … (21r) Prim. Hymnus (AH 51, Nr. 55). – (21v) Ps 6. – (22v) Antiphona. Miserere mei domine quoniam infirmus sum sana me domine quoniam conturbata sunt ossa mea. – Capitulum. – Res ponsorium mit Versikel. – (23r) Oracio. Omnipotens sempiterne Deus confitentibus tibi famulis tuis pro tua pietate relaxa peccata … (23v) Terz. Hymnus (AH 51, Nr. 55). – (24r) Ps 32. – (24v) Antiphona. Delictum meum cognitum tibi feci et iniusticiam meam a te non abscondi. – Capitulum. – (25r) Responsorium mit Ver sikel. – Oracio. Exaudi quesumus domine preces supplicum et confitencium tibi … (25r) Sext. – (25v) Hymnus (nur Verweis: AH 51, Nr. 55). – Ps 38. – (27r) Antiphon (vgl. Cantus Nr. 204374). – Capitulum. – Responsorium mit Versikel. – Oracio. Deus infinite misericordie veritatisque immense propiciare iniquitatibus nostris et animarum nostrarum … (27v) Non. Hymnus (nur Verweis: AH 51, Nr. 55). – Ps 130. – (28r) Antiphon (CAO Nr. 1466). – Capitulum. – Responsorium mit Versikel. – (28v) Oracio. Adesto domine supplicacionibus nos tris ne sit a nobis famulis tuis longinqua tua miseracio … (28v) Vesper. – (29r) Ps 102,2–19. – (30r) Antiphon (CAO Nr. 7820o). – Capitulum. – Hymnus (AH 51, Nr. 89). – (30v) Magnificat (nur Verweis). – Antiphon (CAO Nr. 5013). – Oracio. Deus sub cuius oculis omne cor trepidat omnesque conscientis (!) contremiscunt … (31r) Komplet. Ps 102 (nur Verweis). – Antiphona. Non avertas faciem tuam a me et similis ero descendentibus in lacum. – Hymnus (AH 51, Nr. 22). – (32r) Capitulum. – Versikel. – Canticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 4461). – (32v) Oracio. Fideles tuos quesumus domine ab ira tua confugientes ad te paterna recipe pietate … – (33r) Alia oracio. Omnipotens sempiterne Deus ne nos miseros peccatores permittas propter peccata nostra perire … (33v) ganzseitige Miniatur (s. bei A). 3 (34r–51r) Cursus pro defunctis. Hymnen (AH 30, Nr. 81 Vesper und Completorium vertauscht). Tit.: Cursus pro defunctis fidelibus animabus. (34r) Invitatorium mit Verweis auf Ps 95. – Matutin. Hymnus. – (34v) Ps 3. – (35r) Antiphon (vgl. CAO Nr. 1512). – Benediccio. Exaudi domine preces servorum tuorum ut anime fidelium defunctorum a penis liberentur infernorum … – (35v) Leccio prima. Inc.: Deus cui omnia mo riencia vivunt et cui non pererint (!) moriendo corpora nostra sed mutantur in melius … – Expl.: … iudicare vivos et mortuos et seculum per ignem. Tu autem domine. – (36r) Responsorium mit Versikel. – Leccio secunda. Inc.: Deus vite dator humanorum corporum reparator qui a peccato ribus exorari voluisti exaudi quesumus preces nostras … – Expl.: … per resurgentem a mortuis dominum Ihesum Christum. Tu autem. – (36v) Responsorium mit Versikel. – Leccio tercia. Inc.: Te domine sancte pater omnipotens eterne Deus supplices deprecamur pro animabus omnium fidelium defunctorum quas ab origine mundi accersiri voluisti … – Expl.: … per resurgentem a mortuis dominum nostrum Ihesum. Tu autem. – (37r) Responsorium mit Versikel. Parallelüberlieferung der Lectiones: Innsbruck, ULB Tirol, Cod. 398, Bl. 91v–93r. (37v) Laudes. Ps 51,3–21. – (38v) Antiphon (CAO Nr. 1912). – (39r) Capitulum. – Hymnus. – (39v) Ps 72,18 (nur Verweis). – Antiphon (Cantus Nr. 200808). – (40r) Oracio. Respice quesu mus omnipotens et misericors Deus super animas omnium fidelium defunctorum … (40r) Prim. Hymnus. (40v) Ps 67. – (41r) Antiphon (CAO Nr. 2444). – Capitulum. – Responso rium mit Versikel. – (41v) Oracio. Da quesumus omnipotens et misericors Deus ut anime omnium 316 Cod. 654 fidelium defunctorum hic et ubique in Christo quiescencium … (41v) Terz. Hymnus. – (42r) Ps 56. – (43r) Antiphon (CAO Nr. 1211). – Capitulum. – Responso rium mit Versikel. – (43v) Oracio. Sacratissimam domine dilectissimi filii tui passionis memo riam devote recolentes pro liberacione animarum … (44r) Sext. Hymnus. – Ps 57. – (45r) Antiphon (vgl. Cantus Nr. g02393). – Capitulum. – Respon sorium mit Versikel. – (45v) Oracio. Deus qui unicum filium tuum ut servum redimeres sponte in mortem crucis tradidisti … (45v) Non. – (46r) Hymnus. – Ps 130. – (46v) Antiphona. Domine Ihesu Christe pastor bone qui ovem perditam reportasti ad ovile veniant omnes sancti in adiutorium animabus omnium fidelium qui pius et misericors es salvator noster. – Capitulum. – Responsorium mit Versikel. – (47r) Oracio. Domine Ihesu Criste fili Dei vivi qui precioso sanguine tuo mundum redemisti … (47v) Vesper. Ps 116,1–9. – (48r) Antiphon (vgl. CAO Nr. 1783,1). – Capitulum. – Responso rium mit Versikel. – (48v) Hymnus. – (49r) Versikel. – Magnificat (nur Verweis). – Antiphon (CAO Nr. 4689). – Magnificat (nur Verweis). – Oracio. Domine Ihesu Criste iudex vivorum et mortuorum qui ob hoc irasceris ut subvenias ad hoc minaris … (49v) Komplet. Ps. (recte: Ies. 38,10). – (50r) Antiphon (Cantus Nr. g01573). – Hymnus. – (50v) Capitulum. – Versikel. – Canticum Simeonis (nur Verweis). – Antiphon (CAO Nr. 3732). – Ora cio. Deus qui unigeniti filii tui domini nostri Ihesu Christi precioso sangwine humanum genus redimere dignatus es … (51v) ganzseitige Miniatur (s. bei A). 4 (52r–66r) Septem psalmi Paenitentiales mit Allerheiligenlitanei. (52r–59v) Tit.: Septem psalmi penitenciales sequuntur ut infra (Ps 6, 32, 38, 51, 102, 130, 143). Nach jedem Psalm jeweils Gloria. Anschließend Antiphon (CAO Nr. 3861). (59v–64r) Litania omnium sanctorum. (64r–66r) Orationes. (64r) Inc.: Et veniat super nos misericordia tua domine … – (64v) Alia oracio. Omnipotens sem piterne Deus dirige actus nostros in beneplacito tuo … – Oracio. Fac nos quesumus domine sancte Marie semper virginis subsidiis attolli … – (65r) Oracio. Ecclesie tue quesumus domine preces placatus admitte … – Oracio. Pretende quesumus domine famulis et famulabus tuis … – (65v) Oracio. Suppliciter te Deus pater omnipotens qui es creator omnium rerum … – Oracio. Ure igni sancti spiritus renes nostros et corda nostra domine … – (66r) Oracio. Animabus que sumus domine omnium fidelium defunctorum oracio proficiat … (66v) ganzseitige Miniatur (s. bei A). 5 (67r–90v) Vigiliae mortuorum. Tit.: Vigilie mortuorum sequuntur. – Anrufung: Oremus pro fidelibus defunctis. Requiem eternam dona eis domine. (67r) Invitatorium mit Ps 95. – (68r) Ps 5. – (69r) Antiphon (CAO Nr. 2244). – Ps 6. – (70r) An tiphon (CAO Nr. 1921). – Ps 7. – (71v) Antiphon (CAO Nr. 3875). – Versikel. – Leccio prima (Iob 7, 16–21). – (72r) Responsorium mit Versikel. – Leccio secunda (Iob 10,1–7). – (72v) Res ponsorium mit Versikel. – Leccio tercio (!) (Iob 10, 8–12). – (73r) Responsorium mit Versikel. (73v) Tit.: In secundo nocturno. Ps 23. – (74r) Antiphon (CAO Nr. 3250). – Ps 25. – (75v) An tiphon (CAO Nr. 2146). – Ps 27. – (76v) Antiphon (CAO Nr. 1948). – Leccio quarta: Iob 13, 23–28. – (77r) Responsorium mit Versikel. – (77v) Leccio quinta (Iob 14,1–6). – (78r) Respon sorium mit Versikel. – Leccio sexta (Iob 14, 13–16). – (78v) Responsorium mit Versikel. – Ps 6 (nur Verweis). – Antiphona. Non derelinquas me domine Deus meus ne discesseris a me. – Ps 41. – (80r) Antiphon (CAO Nr. 4696). – Ps 42. – (81r) Antiphona. Sitivit anima mea ad Deum fontem vivum quando veniam et apparebo ante faciem domini. – Versikel. – Leccio septima (Iob 17). – (81v) Responsorium mit Versikel. – (82r) Psalmus (recte: Lectio Iob 19,20–27). – (82v) Responsorium mit Versikel. – (83r) Leccio VIIIo (Iob 10,18–22). – (83v) Responsorium mit Ver Cod. 654 317 sikel. – Ps 51 (nur Verweis). – Antiphon (CAO Nr. 2810). – Ps 65. – (84bv) Antiphon (CAO Nr. 2767). – (85r) Antiphon (CAO Nr. 3725). – Psalmus (recte: Lectio: Is 38,10–20). – (86r) Antiphon (CAO Nr. 2674). – Ps 148 (nur Verweis). Antiphon (CAO Nr. 4154). – Canticum Za chariae (nur Verweis). – Antiphon (CAO Nr. 2601). – (86v) Oracio. Inclina domine aurem tuam ad preces nostras quibus misericordiam tuam suppliciter deprecamur ut animam famuli tui … – (87r) Alia oracio. Quesumus domine pro tua pietate miserere anime famuli tui a contagiis morta litatis … – Oracio. Deus qui nos patrem et matrem honorare precepisti miserere queso clementer animabus patris mei et matris mee omniumque parentum … – Oracio. Deus venie largitor et humane salutis amator quesumus clementiam tuam … – (87v) Oracio. Fidelium Deus omnium conditor et redemptor animabus famulorum famularumque omnium fidelium … – (88r) Antiphon (CAO Nr. 3732). (88r) Tit.: Vespere mortuorum. Ps 116, 1–9. – (88v) Antiphon (CAO Nr. 4293). – Ps 120 (nur Verweis). – Antiphon (CAO Nr. 3038). – Ps 121. – (89r) Antiphon (CAO Nr. 2402). – Ps 130 (nur Verweis). – (89v) Antiphon (CAO Nr. 4899). – Ps 138. – (90r) Antiphon (CAO Nr. 4159). – Ma gnificat (nur Verweis). – Antiphon (CAO Nr. 1211). – Antiphon (CAO Nr. 4115). (90v–93r) Oratio pro animabus (Ablassgebet Papst Johannes’ XXII.). (90v) Ablassvermerk (rot): Oracio pro animabus. Papa Iohannes vicesimus secundus concessit cuilibet dicenti subscriptam oracionem cum devocione in cimiterio cum uno Pater noster Ave Maria et Credo tot annos indulgenciarum quot corpora requiescunt et Innocencius papa tercius concessit cuilibet dicenti pro quolibet vice trecentos annos indulgenciarum. (91r) Avete omnes fideles anime in sancta pace quiescite qui vos redemit suo precioso sanguine vobis dignetur misereri penasque vestras … – (91v) Ps 130 (nur Verweis). – Oremus. Miserere domine per sanctam anuncciacionem adventum nativitatem ieiunium … – (92v) Oracio. Fide lium Deus omnium conditor et redemptor animabus famulorum famularumque tuarum omnium fidelium … Darunter Ablassvermerk (rot): Ista metra subscripta sunt dicenda in cimiterio et qui ea devote dixerit ab aliquibus summis pontificibus maximas consequitur indulgencias ut sequuntur. (schwarz:) Sex milia sexcenta sexaginta sex tot fertur Ihesus pro nobis vulnera passus. Da re quiem cunctis Deus hic et ubique sepultis. Ut sint in requie propter vulnera quinque. (93v) ganzseitige Miniatur (s. bei A). 6 (94r–98v) Anfänge der vier Evangelien. (94r) Tit.: Quatuor inicia ewangeliorum. – (94r) Tit.: Secundum Matheum. Matth 1,1–16. An schließend: Deo gracias per istos sermones et per verba et virtutem sancti ewangelii dignetur dominus noster Ihesus Christus nos et fructus terre nostre custodire … – (95v) Oracio. A domo tua quesumus domine spiritales nequicie repellantur et aeriarum discedat maligni … – Tit.: Se cundum Marcum. Marc 1,2–8. – (96v) Tit.: Secundum Lucam. Luc 1,5–17. – (97v) Tit.: Secun dum Iohannem. Ioh 1,1–14. Anschließend: Deo gracias per istos sermones et per verba et per virtutem sancti ewangelii dignetur dominus noster Ihesus Christus … – (98v) Verweis: Oracio ut supra. (99r–v) nur Rahmung und Linierung. 7 (100r–108v) ‹Ps.-›Birgitta Suedensis: Orationes XV de passione dni (Druck: GW 4362–4383). Tit.: Incipiunt quindecim oraciones sancte Brigite. – Inc.: O domine Ihesu Criste eterna dulcedo te amancium iubilus excedens omne gaudium et omne desiderium … – Expl.: … ut post vite mee terminum te laudare merear cum omnibus sanctis tuis in eternum Amen. Ave dulcissime Ihesu Christe. (109r) leer. (109v) ganzseitige Miniatur (s. bei A). 318 Cod. 691 8 (110r–115v) Oratio pro BMV (Ablassgebet Papst Innozenz’ IV.). (110r) Ablassvermerk (rot): Hanc oracionem suprascriptam composuit s. Bonaventura quam postea Innocencius papa quartus confirmavit et dedit omnibus eam devote dicentibus de quolibet vice trecentos dies indulgenciarum. (110r) Domina mea sancta Maria perpetua virgo virginum mater summe benignitatis et miseri cordie deprecor te … Parallelüberlieferung: Innsbruck, ULB Tirol, Cod. 402, Bl. 159r. (116r) nur Rahmung und Linierung. (116v) ganzseitige Miniatur (s. bei A). 9 (117r–202r) Itinerarium seu peregrinatio BMV (Druck: GW M15411 und M15415). Tit.: Prefacio in itinerarium seu peregrinacionem: beate virginis et Dei genitricis Marie. – Inc.: Subsequens oratio de beata virgine et Dei genitrice Maria continens principaliora ipsius et filii eius gesta … – Expl.: … ac te sibi congregare constituit et previdit in eternum. Amen. (rot:) Finis itinerarii seu peregrinacionis beate Marie virginis. (122r–v, 132r–v, 143v–144r, 153v–154v, 163v–165r, 177v–178v, 187v–188r) ganzseitige Minia turen (s. bei A). (202r) Schlussschrift mit Schreibernennung und Datierung: Hec oraciones scripte sunt per me Georgium Höltzl de oppido Matikofen in expensis domini Caspar Newhauser tunc temporis iudex in Clusna in anno domini millesimo quadragentesimo nonagesimo sexto vigesimo octavo die mensis Iulii etc. Laus Deo (28. Juli 1496) (Colophons Nr. 5053). (202v) nur Rahmung und Linierung sowie Stempel (s. bei G). (203r) nur Rahmung und Linierung. (203v und Ir–v) leer. U. S. Cod. 691 ARGUMENTA LIBRORUM DICTYS CRETENSIS Pap. 5 Bl. 210 × 145. Neustift (?), Anfang 16. Jh. B: Pap. am unteren Blattrand fleckig (Wasserschaden). Wasserzeichen nur fragmentarisch erhalten, kein Nach weis möglich. Lage: (1+II)5. Bl. 1 eingehängtes Einzelbl., am Spiegel des VD angeklebt. Am oberen Blatt rand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 165/170 × 95, seitlich von Blindlinien begrenzt, zu 30–32 Zeilen. Humanistische Kursive von zwei Händen: 1) Bl. 1r–4r; 2) Bl. 4v. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Am VD Signaturschild (s. bei G). Spiegel Pap. Am Spiegel des VD Exlibris, Signaturen und Stempel (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II1 D 5 (Tinte), II3 E 27 S. 2 (Bleistift) und 691 (Tinte). Am VD gelbes Signaturschild der ULB Tirol 691. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Am Spiegel des VD Stempel der Soprintendenza Tridentina 20. Jh. L: W. Speyer, Die unbekannte Epitome des Dictys Cretensis im Codex Brixinensis 691. Rheinisches Museum für Philologie, N. F. 107/1 (1964) 76–92. – Kristeller I 439. – Katalog ULB Tirol VII 470, 485. Argumenta librorum Dictys Cretensis: De Troiani regni excidio cum appendiculis (Ed. Speyer 84–91 [basierend auf Neustift, Stiftsbibl., Cod. 691]). Cod. 694 319 Inc.: Minos Iovis et Europe filius Androgeum Ariadnam Phedram Daucalionem (!) et Molum p rogenuit. Ex Deucalione Idomeneus ex Molo Meriones seu Mirion et Idomeneus alter nascitur. Item Tantalus Iovis filius genuit Pelopem … – Expl.: … ex quo Anchises editus est Aeneam filium procreavit. Der Text umfasst Inhaltsangaben der sechs Bücher des Dictys Cretensis, die von zwei Appendices gerahmt werden: (1r) Appendix prima. Ohne Tit. – (1v) Tit.: Argumentum primi libri. – (2v) Tit.: Argumentum secundi libri. – (3r) Tit.: Argumentum tercii libri. – (3r) Tit.: Argumentum quarti libri. – (3v) Tit.: Argumentum quinti libri. – (4r) Tit.: Argumentum sexti et ultimi libri Dictis Cretensis. – (4v) Tit.: De origine Troiano rum (= Appendix secunda). Zur Überlieferungsgeschichte sowie zum Aufbau des Textes vgl. Speyer 76–83. (5r–v) leer. A. P. / U. S. Cod. 694 PAULUS URSINUS Pap. 27 Bl. 205 × 150. Tirol, 1523. B: Bilder der Wasserzeichen s. WZMA. Lagen: 2.VI24 + (II–1)27. Gegenbl. zu Bl. 25 als Spiegel auf dem HD aufgeklebt. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 140/145 × 70/75, seitlich von Blindlinien begrenzt, zu 23–26 Zeilen. Humanistische Kursive, dat. (erschlossen) 1523. Vermutlich Autograph (Widmungsexemplar). A: Weinrote Marginalien, Personenangaben im Text, Überschriften und Schlussschrift. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Pap. leicht beschädigt, Bindung gelockert (Lage 1 oben gelöst). Am Rücken bzw. VD Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel des VD und HD Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] E […] (Tinte, radiert), II3 E 30 S. 2 (Bleistift) und 694 (Tinte). Am Rücken bzw. VD gelbes Signaturschild der ULB Tirol 694. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Ger main an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 137. – Kristeller I 439. – Katalog ULB Tirol VII 473, 485. Paulus Ursinus: Dialogus de formidine diluvii instantis anni vigesimi quarti abigenda. (1r) Tit.: Dialogus de formidine diluvii instantis anni vigesimi quarti abigenda. (1v) Widmungsbrief. Tit.: Epistola. – Inc.: Reverendissimo in Christo patri ac domino domino Se bastiano Dei gracia episcopo Brixinensi necnon egregiis eximia doctrina et sapiencia eiusdem ecclesie prelatis ac canonicis Paulus Ursinus philosophus et medicus s. d. Que vicio nostro non proveniunt, queque in nostra minime sita sunt … – Expl.: … complurium interventu beneficiorum me frui cercior sum. (3r) Text. Tit.: Dialogus non minus festivus quam facetus de formidine diluvii anni 24ti ultra sesquemil lesimum abigenda. Interlocutores: Philagrus Mercurius Genius. – Inc.: Phil[agrus]. Ut quid Mercuri hic ades galero redimitus et citharam gestans … – Expl.: … si non suprema de omnibus iudicet dies. Finis. (27v) leer. Paulus Ursinus ex Nisia (Neiße) wurde nach dem Studium in Bologna und Ferrara 1512 zum Doktor der Medizin promoviert (vgl. C. Zonta, Schlesische Studenten an italienischen Universitäten. Stuttgart 320 Cod. 726 2004, 168). Seine dem Brixner Fürstbischof Sebastian Sprenz (1521–1525) gewidmete Schrift steht im Kontext der Sintflutdebatte, die durch astrologische Vorhersagen für das Jahr 1524 ausgelöst wurde. Zur Debatte s. H. Talkenberg, Sintflut. Prophetie und Zeitgeschehen in Texten und Holzschnitten astro logischer Flugschriften 1488–1528. Tübingen 1990. L. Š. Cod. 726 RITUALE NEOCELLENSE Pap. III, 78, I Bl. 120 × 100. Neustift, 1523 (mit Zusätzen 16. und 17. Jh.). B: Bilder der Wasserzeichen s. WZMA. Lagen: (II–2)II + 10.IVIII,77(78) + (I–1)I. Gegenbl. zu Bl. I herausge schnitten, Gegenbl. zu Bl. II als Spiegel auf dem VD, Gegenbl. zu Bl. I als Spiegel auf dem HD aufgeklebt. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 24 und 37 doppelt an geführt, Bl. 77 nicht foliiert. S: Bl. 1r–70v: Schriftraum 95 × 70, zu 21 Zeilen, seitlich und tw. oben von Tintenlinien gerahmt. Textualis mit kursivem Einschlag, dat. 1523 (Bl. 24v, 40v, 65r, 70r und 70v [11.6.]). Schreiber: Franciscus Prensteiner (Bl. 70v), Dekan von Neustift, vgl. Giner 12. Bl. 71r–73v: Schriftraum 105 × 70, zu ca. 18 Zeilen, seitlich von Tintenlinien gerahmt. Humanistische Halb kursive in schwarzer und roter Tinte 16. Jh. Bl. 74r: Schriftraum 100 × 70, zu 10 Zeilen, seitlich von Tintenlinien gerahmt. Antiquakursive 17. Jh. (1684?). Bl. 76r: Schriftraum 105 × 70, zu 14 Zeilen, seitlich von Tintenlinien gerahmt. Antiquakursive 16. Jh. (1581?). A: Bl. 1r–70v: Rote Auszeichnungsstriche, Überschriften, Benediktionskreuze (tw. gerahmt), Satzmajuskeln und zweizeilige Lombarden mit Punktverdickungen. Repräsentanten. Bl. 71r–73v: Weinrote Überschriften und Satzmajuskeln. E: Renaissance-Einband: hellbraunes Leder mit Blinddruck über Holz. Brixen oder Neustift, 2. H. 16. Jh. VD und HD gleich: zwei aus je dreifachen Streicheisenlinien begrenzte Rahmen: Rahmen 1: Rolle Köpfe in Medaillon, dazwischen Rankenmuster und Inschrift SOLI bzw. CARO bzw. OCDA (Nr. 1 = Neuhauser, Rolleneinbände, R 56 [Gruppe 7]); Rahmen 2: Rolle S-förmiges (Schrift-?)Band, ineinander verhakt (Nr. 2). Im Mittelfeld Einzelstempel Maria mit Kind (Nr. 3), von mehrfachen Streicheisenlinien begrenzt. Reste einer Leder-Metall-Schließe. Kanten gerade. Rücken: blau gefärbt (wie öfters in Neustift), drei Doppelbün de. Im ersten und zweiten Feld Papiertitelschild MS Manuale Benedictionum ac Rituum Neocellensium, im dritten Signaturschild (s. bei G). Kapitale mit rötlich-weißem Zwirn umstochen. Spiegel Pap. 16. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signatur der ULB Tirol II3 F 12 S. 2 (Bleistift) und 726 (Tinte). Bl. Iv Abklatsch des Stempels der ULB Tirol 19. Jh. (urspr. möglicherweise auf Recto des herausgerissenen Gegenbl. zu Bl. I). Am Rücken gelbes Signaturschild der ULB Tirol 726. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. IIIv Stempel der Soprintendenza Tridentina 20. Jh. L: Gschwend 86–88 (ohne Nennung der Sign.). – Lipphardt III 1086; VI 352, Nr. 629 (Sigle Neust4). – Stefani 117. – Peintner, Stiftsbibliothek 126. – Gozzi 548, Nr. 80. – Katalog ULB Tirol VIII 127, 529. (Ir–IIIv) leer bis auf Stempel (s. bei G). 1 (1r–70v) Rituale Neocellense. Vgl. Cod. 194 und Sign. 14820. (1r–7r) In purificatione BMV. (1r) Benedictio candelarum. Tit.: Purificacionis Marie cereorum benedictio post 3as. – Inc.: O remus. Erudi quesumus domine plebem tuam … – (5v) Processio candelarum. – (6r) Canticum Zachariae. Cod. 726 321 (7r–9r) Fer. 4 cinerum (nicht in Cod. 194 und Sign. 14820). (7r) Benedictio cinerum. Tit.: Feria IIIIa. In capite ieiunii. Benedictio cineris. – (8v) Impositio cinerum. Rubrik: Tunc ponendi sunt cineres super capita singulorum dicens. (9r–21v) Dnca in palmis. (9r) In die palmarum ad processionem. – (12v) Exorcismus florum et frondium. – (12v) Benedic tio palmarum. – (16v) Praefatio. – (17v) Aspersio et thurificatio palmarum. – (18r) Distributio palmarum. – (18v) In reditu. – (19v) Adoratio crucis. – (21r) Canticum puerorum. – (21v) In reditu ante altare oratio. (21v–24v) Fer. 5. in coena dni. (21v) In cena domini benedictio ignis. – (23r) Ad mandatum. (24v) Datierung (in Rot) 1523. (25r–35r) Fer. 6. in parasceve. (25r) Adoratio crucis. – (29r) Depositio crucis (Gschwend 86). – (29v) Vesper. (35r–44r) In vigilia paschatis. (35r) Benedictio ignis. – (36v) In reditu canitur himnus Prudencii (AH 50, Nr. 31). – (37r) Ex sultet. – (39v) Benedictio aquae. – (43v) Litaniae sanctorum. (40v) Datierung 1523. (44r–46v) Dnca resurrectionis dni (Gschwend 86–88). (46v–47r) Benedictio aquae in vigilia Pentecostes (Verweis). (47r) In assumptione BMV. (47r–55v) Ordo paenitentiae. (55v–58r) Einkleidung der Novizen. Tit.: De induitione noviciorum. (58r–65r) Profess. Tit.: De professione noviciorum. – (65r) Datierung 1523. (65r–67r) Ordo für die Aufnahme von Laien. Tit.: De fraternitate laicali. (67r–70v) Totenliturgie für die Konventualen. Tit.: De obitu fratrum professorum nostri ordinis. Mit Datierung Bl. 70r 1523. (70v) Schlussschrift mit Datierung und Schreibernennung: 1523 f. p. (= Franciscus Prensteiner). Anno domini MoCCCCCXXIII tercio Idus Iunii is finitus est codiculus seu Obsequiale. Scriptus ad honorem et gloriam unigeniti ante secula summi patris filii domini nostri Hiesu (!) Christi eiusdemque matris gloriose virgini (!) Marie Novecelle patrone graciosissime per fratrem Fran ciscum Prenstainer tunc temporis in Novacella decanum (11. Juni 1523). 2 (71r–73v) Ordo baptisterii (vgl. J. Baur, Die Spendung der Taufe in der Brixner Diözese in der Zeit vor dem Tridentinum [Schlern-Schriften 42]. Innsbruck 1938, 35–36). Tit.: Exorcismus salis minor ad baptizandum si alias non esset in promptu. – Inc.: Exorciso te creatura salis in nomine patris omnipotentis et in charitate domini nostri Ihesu Christi et in vir tute spiritus sancti … – Expl.: … sit tibi Deus in vitam eternam. Pax tecum. 3 (74r) Verfügung des Propstes Fortunat Troyer (Propst 1678–1707), betreffend die Offizien beim Begräbnis eines Klerikers, dat. 4. August 1687. 322 Cod. 748 Tit.: Declaratio Reverendissimi et Domini Domini Prælati Fortunati circa Sepulturam Clerici. (74v–75v) leer. 4 (76r) Vermerke über den 1581 verstorbenen Neustifter Propst Augustinus Schbäl (recte: Schabl) und den 1581 zum Propst gewählten Adamus Lang. (76v–77v) leer. (Ir–v) leer. W. N. / U. S. Cod. 748 MEDIZINISCHE SAMMELHANDSCHRIFT Pap. III, 220 Bl. 290 × 215. T. I–III: Wiener Raum (?), um 1440, T. IV: Schwäbisch-Schweizer Raum, um 1430. B: Vier Teile: I (III, 1–40), II (41–52), III (53–137), IV (138–219[220]). Wohl im Zuge der Bindung am oberen rechten Blattrand zeitgenössische Lagenzählung jeweils am Lagenbeginn, tw. ausgeschrieben primo, tw. in arabischen Ziffern 2–9 und 12–19, tw. in römischen Zahlen X–XI ergänzt. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 211 doppelt angeführt. E: Schmuckloser gotischer Einband: Leder über Holz. Tirol (?), Mitte 15. Jh. Der urspr. Bezug aus rotgefärbtem Leder auf den Deckeln nur in Spuren vorhanden, tw. noch als Einschläge auf der Innenseite der Deckel sichtbar. Die urspr. unter dem Bezug aufgeklebten Pergamentstreifen auf der äußeren und inneren Seite der Deckel freiliegend. Am Holz tw. Einritzungen (Rautenmuster) zur besseren Haftung sichtbar. Am VD und HD Spuren von je fünf Beschlägen. Spuren von zwei Langriemen-LederMetall-Schließen. Kanten abgeschrägt und gerade. Rücken: nach der Entfernung des Lederbezuges und der Beschläge mit braunem Leder neu bezogen, drei Doppelbünde. Kapitale mit Zwirn umstochen (oben urspr. dunkel gefärbt, unten naturfarben), gebrochen. An der Innenseite der Deckel Holz freiliegend. An der Innen seite des VD Exlibris und Signatur (s. bei G), an der Innenseite des HD Spuren des entfernten beschriebenen urspr. Spiegels. G: Geschenk des Neustifter Präbendars und Klosterarztes Bertoldus de Saxonia ans Stift (gemeinsam mit Cod. 123 und vermutlich auch dem tw. von denselben Händen geschriebenen Cod. 453): Bl. IIIv Schen kungsvermerk (s. Nr. 2) mit Neustifter Wappen, Bl. 137v Besitzereintragung Bertoldus medicus Saxo. Zu Bertoldus vgl. Liber anniversariorum Novae Cellae Brixinensis (MGH Necrologia Germaniae III 46): „Item magister Bertoldus de Saxonia, medicine doctor, prebendarius et phisicus huius monasterii, in vita sua multa fecit bona et libere omnia sua dedit monasterio, prout continetur in litera prebendali, quam monasterio dedit“. Bl. 137v Besitzvermerk 15. Jh. Liber iste est monasterii sancte Marie virginis in Novacella Brixi nensis dyocesis mit Neustifter Wappen, Bl. 219r Besitzvermerk 15. Jh. (von gleicher Hand) Iste liber est monasterii sancte Marie virginis in Novacella. Bl. 1r Inhaltsangabe und Besitzvermerk 18. Jh. Summarium de febribus per Antonium Gaynerium. In fine Ysac de urinis. Ad Bibliothecam Neocell. An der Innenseite des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. Ir alte Signaturen der ULB Tirol II`3 B 14 (Tinte, radiert), II`4 C 11 (Bleistift) und 748 (Tinte). Letztere beiden auch auf Bl. IIIv (Bleistift), 748 auch an der Innenseite des VD. Am Rücken gelbes Signaturschild der ULB Tirol 748. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Kristeller I 439. – Neuhauser, Neustift 89. – Katalog ULB Tirol VIII 247, 530. I (III, 1–40) B: Pap. tw. stark verschmutzt (u. a. Wasserschaden), an den Rändern leicht eingerissen, Bl. 7 Loch. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: Neustift, Stiftsbibl., Cod. 748, T. III. Lagen: 3III (Vorsatzbl.) + (VI+1)13 + VI25 + (VII+1)40. Bl. 1 und 39 ein Cod. 748 323 geklebte Einzelbl. Lagen außen und in der Mitte durch Pergamentfälze verstärkt. Am oberen Blattrand mittig Kustoden in arabischen Ziffern 1–3. S: Bl. Ir–IIIv (Vorsatzbl.) horizontale Tintenlinienrahmung, Bl. Iv–IIr zusätzlich seitlich von zwei Tintenlinien begrenzt. Schriftraum 210/225 × 130/140, seitlich von Tintenlinien begrenzt, zu 45–49 Zeilen. Bastarda (= Schreiber von T. II und III, von Neustift, Stiftsbibl., Cod. 453, T. II–IV und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Bl. Ir–v, 1r, 40v vermutlich deutlich spätere Nachträge vom selben Schreiber. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen, Bl. 29v und 31v jeweils rote Überschrift am Blattrand. Platz für Lombarden freigelassen, nicht ausgeführt. Tw. rote, tw. schwarze Repräsentanten. Bl. IIIv Neustifter Wappen in schwarzer Tinte. 1 (Ir–v) Rezepte und Notizen. (Ir) 1) Inc. mut.: ] aqua de floribus fabarum aut aqua furfuris aut aqua … – Expl.: … cum istis tribus aquis maxime albatur facies. 2) Tit.: Contra fluxum sanguinis narium. – Inc.: Sanguinaria id est bursa pastoris sive succus … – Expl.: … in albumen ovi involvatur et immittatur. (Iv) 3) Inc.: Signa generalia putridarum eis propria sunt duo ut dicit Averroes … – Expl.: … Si non tunc est effimera et hoc supposito quod febret. 4) Tit.: De tertiana continua. – Inc.: Nota. Galieus (!) 2o de crisi dicit quod si videris urinam egri tendentem … – Expl.: … pervenerint VII periodi deinde consumetur. 5) Inc.: Ab infirmitate surgentibus cibaria nociva petentibus non est denegandum … – Expl.: … potius natura viget dum vota replentur. 6) Inc.: Vehemenciores fiunt motus egrium (!) et acta eorum in nocte … – Expl.: … quando somp num auferunt ut in nocte. (IIr–IIIr) tw. Rahmung. 2 (IIIv) Schenkungsvermerk mit Inhaltsangabe: Liber iste donatus est monasterio sancte Marie virginis ad Novamcellam Brixinensis dyocesis a dive memorie magistro Bertoldo de Saxonia doctore medicine eximio cum aliis libris et rebus pro prebenda et remedio anime sue. Obiit 1476. Continet vero liber presens summarium de febribus editum per Antonium de Gaynerys. Librum Ysaac de urinis. Item librum Ysaac de febribus. Synonima Rasis. 3 (1r) Verse und Notizen. 1) Am oberen Blattrand sechs Verse aus Regimen Sanitatis Salernitanum (vgl. S. Renzi, Col lectio Salernitana V. Naples 1859, V. 1931–1937). Inc.: Triplex causa febrem generat custodit et auget … – Expl.: … quare generatur putrida febris. 2) Inc.: Nota. Decrementa et incrementa paroxismorum ad invicem aut sunt secundum quan titatem aut secundum tempora aut secundum qualitatem. Si secundum quantitatem aut lon gior … – Expl.: … aut non precesserunt et sic est principium. 3) Inc.: Nota. Anticipacio paroxismi augmentum non significat nisi cum hoc longitudo temporis aut fortitudo febris augeatur … – Expl.: … completur digestio. 4 (1v–40v) Antonius Guainerius: Summarium de febribus (Druck: u. a. GW 11586. – Thorndike– Kibre 318). Tit.: Incipit summarium de febribus editum per me Anthonium de Gayneriis ad insignem arcium et medicine doctorem magistrum Antonium Maglianum de Cherio illustrissimi domini domini ducis Subaudie phisicum ad laudem Dei patris et filii et spiritus sancti feliciter incipit. (1v) Prolog mit anschließender Kapitelübersicht. Inc.: [T]ui amoris gratia my Antoni Magliane ad hunc deveni locum … – Expl.: … me venire permittat qui est de virgine natus. (2r) Text. Inc.: [C]um mee sit intencionis hoc in summario febrium … – Expl.: … sic breviter dicta sufficiant que huic summario finem imponit pro quo Ihesu crucifixo sint infinite laudes Amen. 324 Cod. 748 Schlussschrift: Et sic est finis tractatus de febribus ad insignem arcium et medicine doctorem magistrum Anthonium Maglianum de Chero (!) altissimi domini domini ducis Subaudie phisicum editus per Anthonium de Gayneriis de Papia in villa Crambraraci commorantem. 5 (40v) Notizen. 1) Inc.: Confortatio fit multipliciter. Primo per restauracionem bonam sicud spiritus ex vino aro matico dicuntur confortari … – Expl.: … vite ut faciunt bezaartica omnia. 2) Accidens morbi est triplex scilicet preventivus ut rigor concomitans ut dolor capitis et sub sequens ut prostratio appetitus. 3) Nota. Triplex est rigor scilicet precedens et est in interpolates coincidens ex multitudine ma terie et subsequens a fortitudine expulsive. Et isti duo rigores sunt aliquando in compositis febribus. II (41–52) B: Pap. insbes. am Blattrand verschmutzt und eingerissen. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Wasserzeichen: Neustift, Stiftsbibl., Cod. 453, T. III. Lage: VI52. Lage außen und in der Mitte durch Pergamentfälze verstärkt. S: Schriftraum 225/240 × 135/140, seitlich von Tintenlinen begrenzt, zu 51 Zeilen. Links neben dem Schriftraum eine Zählspalte, seitlich von Tintenlinien begrenzt. Bastarda (= Schreiber von T. I und III, von Neustift, Stiftsbibl., Cod. 453, T. II–IV und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). A: Rote Auszeichnungsstriche, Paragraphzeichen, Unterstreichungen und Überschriften. Platz für Lombarden freigelassen, nicht ausgeführt. Repräsentanten. 6 (41r–51v) Christophorus Barsisius: De febrium cognitione et cura (Druck: Clementii Clementi ni lucubrationes. Basileae 1535, 251–283. – Thorndike–Kibre 765, vgl. auch 667). Inc.: [I]ntenciones habende in febribus excerpte ex mente Avicenne ex locis universalibus et par ticularibus. In colerica interpollata pura sunt intenciones (am Blattrand ergänzt: Prima intencio) Est administracio regiminis debiti sex rerum … – Expl.: … Duodecima est respectus et cura accidencium que intelligenti sunt nota. Schlussschrift: Expliciunt declaraciones intencionum in febribus humidarum et earum ordina cionem ad introduccionem iuvenum ad curacionem febrium et contra quosdam detrahentes Avi cenne quod in curis egritudinum insufficienter procedat per magistrum Cristoforum de Pergamo arcium et medicine doctorem collectarum. 7 (52r–v) Tit.: Intenciones morborum materialium. – Inc.: Nota. In morbo materiali sunt habende iste intenciones. Prima amministracio regiminis debiti … – Expl.: … confortatio et remocio male qualitatis morbi etc. 8 (52v) Notizen. 1) Nota. Flebotomia electiva debet fieri parum post ortum solis et post primam et secundam di gestionem et superfluitum expulsionem et post modicum exercicii. Sed coacta potest fieri omni hora secundum necessitatem cogentem. 2) Inter medicinas et corpora currunt proporciones que non comprehenduntur nisi cum experi mento. Cum ergo experieris consequeris quid magis iuvativum sit, ut patet X 3ii capitulo de cura asumatis. III (53–137) B: Pap. leicht verschmutzt, am Blattrand tw. eingerissen. Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung um 1435/40 nach Wasserzeichenbefund. Identische Wasserzeichen: Neustift, Stiftsbibl., Cod. 453, T. III und IV, Cod. 748, T. I. Lagen: (V+1)63 + V73 + 2.VI97 + 4.V137. Bl. 58 eingeklebtes Einzelbl. Lagen Cod. 748 325 außen und in der Mitte durch Pergamentfälze verstärkt. Lage 2–5 am oberen Blattrand mittig Kustoden in arabischen Ziffern. Bl. 137 neuzeitliches oranges Papiersignakel der ULB Tirol. S: Bl. 52r–63v: Schriftraum 205/215 × 145/150. Zwei Spalten, von Bleistiftlinien begrenzt (Bl. 54v–59r und 63v über die zweispaltige Rahmung einspaltig beschrieben), zu 45–51 Zeilen. Bastarda (= Schreiber von T. I und II, von Neustift, Stiftsbibl., Cod. 453, T. II–IV und von Neustift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G). Bl. 91r–137r Überschriften in Textua lis formata. Bl. 69v–73v, 90r und 117v vermutlich deutlich spätere Nachträge vom selben Schreiber. Bl. 91r– 137r Überschriften in Textualis formata. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unter- und Durchstreichungen, Marginalien, Besitzvermerk und Titel. Bl. 132v dreizeilige rote Lombarde. Platz für Lombarden freigelassen, nicht ausgeführt. Repräsen tanten. Rubrikator vermutlich Bertoldus de Saxonia (s. bei G). Bl. 137v Neustifter Wappen in schwarzer Tinte. 9 (53ra–54ra) De aqua vitae (Thorndike–Kibre 779). Tit.: De aqua vite. – Inc.: [H]ec sunt virtutes aque ardentis vel vite. Est enim clarior quam aqua rosacea et quia plus est secure … – Expl.: … distilla cum lento igne. Ista aqua ponitur loco balsami. Parallelüberlieferung: Erfurt, UB, Dep. Erf. CA. 4o 223, Bl. 157v. 10 (54ra–b) Notizen und Rezepte. 1) Inc.: Nota verum secretum in provocacione urine. Recipe corticis interioris … – Expl.: … da cum aqua apericiam. 2) Inc.: Nota oleum factum per decoccionem medicinarum … – Expl.: … per inspissacionem a radiis solaribus facto. 3) Recipe lixinii partis (!) III olei nucum partis (!) I et valet contra cancrum et lupum et ignem persicum in principio certissimum est. 4) (von anderer Hand) Inc.: Antrax et carbunculus quia fiunt ex materia grossa … – Expl.: … fermentum est basis et materia emplastrorum. 5) Inc.: Nota. Quicumque esset lesus vel percussus a malicia Mercurii … – Expl.: … et unge locum et sanabitur. 11 (54v–64v) ‹Raimundus Chalmelli de Vinario›: De pestilentia libri tres, unvollständig (Druck: De peste libri tres opera Iacobi Dalechampii. Lugduni 1553. Ed. [Buch 1]: R. Hoeniger, Der schwarze Tod in Deutschland. Berlin 1882, 160–161, 163–169, 170–176). Tit.: Ista sunt extracta ex quodam libro pestilenciali (rot:) optimo (schwarz:) de pestilencia primo tempore facta. – Inc.: Pestilencie causa dicitur esse triplex. Nam vel est causa radix superior tantum et hec est superior et celestis … – Expl.: … preparatio et earum evacuacio cum laxativis apropriatis dictis vel aliis. Der Text auf Bl. 54v–58r entspricht Ed. Hoeniger 160–161, 163–169, 170–176, der Rest des Textes weicht von der freien Bearbeitung des Dalechampius erheblich ab (vgl. dazu Hoeniger 157–159). Parallelüberlieferung: u. a. Heiligenkreuz, Stiftsbibl., Ms. 485, Bl. 142r; Padova, Biblioteca del Seminario vescovile, Cod. 16, Bl. 1ra. 12 (64v) Rezept. Inc.: Nota. Frequenter fit aurum potabile sic et confert pestilenciatis. Recipe auri optimi … – Expl.: … secundum quos datur credo paucum conferre quia minime digeritur. 13 (64v–66r) Tit.: Dubia quedam optima. – Inc.: Primum dubium quare confert tyriaca et metrida tum cum sint excellentis calefactionis et notum est quod que augmentant caliditatem in corpore nostro … – Expl.: … sub arbore nucis in ortis caulium et sub umbra oleandri. Et sic est finis. 326 Cod. 748 14 (66r) Notiz. Inc.: Nota rupto apostemate et extracta materia … – Expl.: … antracis foliis basiliconis cito implebitur carne. 15 (66v, am oberen Blattrand) Zitat aus ‹Gentilis de Fulgineo›: Consilium de peste, Textbeginn (Druck: u. a. GW 10620). Quoniam gloriosus et excelsus Deus de largitate sua medicinam produxit et medicum velud na ture refugium creavit decet eum nichil negligencie habere in ostendendo sanitatis ingenium ut primo de [ingenio] sanitatis. 16 (66v–69v) Practica contra pestem (Ed.: G. Sticker, Tractatus de epidemia anni 1424. Janus 27 [1923] 15–27. – Thorndike–Kibre 1288). Tit.: Contra pestem alia practica. Schlussschrift: Deo laus eiusque matri virgini Marie, beate Barbare et omnibus sanctis Amen. 17 (69v) Notizen. 1) Nota quod succus peonie vulneribus noviter fissis applicatus nondum ad sanacionem pro venientibus curat. Expertum est. 2) Nota. Elleborus niger decoctus in pomo sub prunis datus in scrupulis II optime purgat t empore pestilencie. 18 (69v–70v) Iohannes de Tornamira: Preservatio et cura apostematum antrosorum pestilentialium (Ed.: K. Sudhoff, Pestschriften aus den ersten 150 Jahren nach der Epidemie des „schwarzen Todes“ 1348 III. Sudhoffs Archiv 5 [1912] 48–53. – Thorndike–Kibre 313). Tit.: Preservacio et cura apostematum antrosorum pestilencialium secundum intencionem Iohan nis de Tornamira. 19 (70v) Rezepte. Tit. (am Blattrand): Receptum preservacionis aliud. – Inc.: Contra aeris infeccionem. Eius enim attraccione membra principalia … – Expl.: … sumendum est ad modum unius nucis mus sine potu. 20 (70v) Rezept. Tit. (am Blattrand): Nota contra antracem. – Inc.: Nota. Tempore pestilenciali si vis ut apostema ex materia venenosa … – Expl.: … postea ligetur et superponatur pannus. 21 (71r) Kurzes Pestregimen. Tit.: Remedia et regimen contra pestilenciam. – Inc.: Oportet ut extrahantur (gestrichen: primo) superfluentes humide primo per flebotomiam … – Expl.: … vitetur ira et tristicia et fiat conver sacio socialis cum dilectis. 22 (71r–72r) Iohannes de Sancta Sophia: Consilium de pestilentia. Tit.: Consilium aliud de pestilencia magistri Iohannis de sancta Sophia. – Inc: Quamvis veneran di domini hec pestis tam admirabilis et horrenda a causis superioribus procedat regulantibus et gubernantibus corpora nostra … – Expl.: … et quia ut cognovi bonum animum geritis erga me. Deus vos conservet ab hac peste Amen. 23 (72r) Rezept. Tit.: Tragea contra pestem. – Recipe tormentille drachmas 15 terre sigilate … fiat tragea. 24 (72r–73v) Tit. (am Blattrand): Contra pestilenciam. – Inc.: Pestilencia est mutacio quedam in aere facta propter admixtionem vaporum corruptorum vel sic est morbus quidam contagi osus … – Expl.: … hoc idem potest fieri si infirmus iaceat continue dormiendo. 25 (74r–90r) Mundinus de Leuciis: Anatomia (Druck: u. a. GW M25667. Weitere Drucke s. A. Cod. 748 327 C. Klebs, Incunabula scientifica et medica. Bruges 1938, ND Hildesheim 1963, 230–231. – Thorndike–Kibre 1306). Tit.: Anathomia Mundini. – Inc.: [Q]uoniam ut ayt Galienus septimo terapentice (!) methodi auc toritate Platonis opus in aliqua sciencia vel arte … – Expl.: … debet esse excarvata ne aggravet pedem. Schlussschrift: Et ex hoc est huius doctrine dictum completum que est de corporis humani anothomia (!) a magistro Mundino artis medicine professore dignissimo ad utilitatem studen cium Bononie compilata. Deo gracias. 26 (90r) Tractatus de anthrace. Tit. (am Blattrand): De antrace. – Inc.: Antrax est apostema quod partim fit de colera partim de sanguine partim de melancholia. De sanguine habet putredinem … – Expl.: … cum bombace ex aliquo ungento calido vel oleo ungatur. 27 (90v–137v) ‹Antonius Guainerius›: Tractatus de peste et venenis (Druck: GW 11591). Tit.: Incipit tractatus de peste ad illustrissimum dominum dominum Philippum Mariam Viceco mitem ducem Mediolanensem Papie Anglarieque comitem et Ianue dominum. (90v) Prolog (Thorndike–Kibre 1036). Inc.: [P]ersepe animadvertens hoc mortale hominum genus iniuriis casibus additum et infinita mortis … – Expl.: … prefati illustrissimi et clementissi mi domini domini mei precipuam utilitatem Amen. (91r) Inhaltsübersicht zu Pars 1. (91r) Pars 1 (Thorndike–Kibre 280). Inc.: [C]um aliquem pestis rapit eum taliter alterat et de vorat ut velud incendium totum in brevi … – Expl.: … eius genitricis alme tociusque celestis curie triumphantis Amen. (122r) Inhaltsübersicht zu Pars 2. (122r) Pars 2 de venenis (Thorndike–Kibre 529). Inc.: [E]t si venenum multipliciter accipi possit hic per venenum id solum intellego quod qualitercumque humano corpori … – Expl.: … dis sonum quod me advertere nichil est meeque compaciendum est ignorancie et non arrogancie imputandum. Schlussschrift: Explicit et est finis. Besitzereintrag und Besitzvermerk (s. bei G). IV (138–219[220]) B: Pap. verschmutzt, am oberen Blattrand tw. Wasserflecken. Bl. 138–147 schmaler (290 × 210). Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lagen: V147 + 6.VI218(219) + 1219(220). Lagen außen und in der Mitte durch Pergamentfälze verstärkt. Am Lagenbeginn Kustoden am obe ren Blattrand in arabischen Ziffern (1 und 3–7) und am unteren Rand tw. ausgeschrieben, tw. in arabischen Ziffern (primus sexternus, 3us sexternus, 4us, 5us, 6tus). Bl. 159v und 207v Lagenreklamanten erhalten. Bl. 138 neuzeitliches gelbes Papiersignakel der ULB Tirol. S: Bl. 138r–212v: Schriftraum 190/195 × 130/135. Zwei Spalten, von Tintenlinien gerahmt, zu 38–43 Zeilen. Zirkelstiche. Bl. 213r–219v: Schriftraum 230/235 × 160/170. Drei Spalten, von Tintenlinien gerahmt, zu 47–53 Zeilen. Bastarda von einer Hand. Ergänzungen und Notizen (Bl. 203r und 209r ausführlich) von demselben Schreiber wie T. I, II und III (= Schreiber von Neustift, Stiftsbibl., Cod. 453, T. II–IV und Neu stift, Stiftsbibl., Cod. 123, Schreiber 2, möglicherweise zu identifizieren mit Bertoldus de Saxonia, s. bei G), Duktus der späteren Nachträge. A: Rote Auszeichnungsstriche, Paragraphzeichen, Unter- und Durchstreichungen, Überschriften. Platz für Lombarden freigelassen, nicht ausgeführt. Repräsentanten. 28 (138ra–161rb) Isaac Iudaeus, ins Lat. übersetzt von ‹Constantinus Africanus›: Liber de urinis (Druck: Opera omnia. Lugduni 1515. J. Peine, Die Harnschrift des Isaac Judaeus, Diss. Leipzig 1919, 10–74). Tit.: Isaac de urinis. 328 Cod. 748 (138ra) ‹Constantinus Africanus›: Praefatio (Thorndike–Kibre 688). Inc.: [I]n Latinis quidem libris nullum auctorem invenire potui qui de urina certam et auctenticam cognicionem dede rit … – Expl.: … de unaquaque urina et ypostasi quid significet etc. (138ra) Text (Thorndike–Kibre 1608). Inc.: [U]rina est colamentum sanguinis et aliorum simili ter humorum de nature quidem actionibus natum … – Expl.: … Urina spumosa in superficie sicut uva (!) dolorem renum significat etc. Schlussschrift: Expliciunt urine Ysac etc. (161v) nur Rahmung. 29 (162ra–212vb) Isaac Iudaeus, ins Lat. übersetzt von ‹Constantinus Africanus›: Liber de febri bus (Druck: Opera omnia. Lyon 1515). Tit.: Incipit Ysaac de febribus. (162ra) ‹Constantinus Africanus›: Praefatio (Thorndike–Kibre 1304). Inc.: [Q]uoniam te karissime filii Iohannes lacrimas mesto cordis dolore perpetim effundere perspexi … – Expl.: … Quibus itaque cognitis curas earum levissime impendere sciet. (162ra) Text (Thorndike–Kibre 820). Inc.: [L]iber ergo iste in V partes dividitur. Prima est de essencia febris eiusque diffinicione febris naturam et causas … – Expl.: … sanguinem qui est eius materia et propter vicinitatem cordis etc. Schlussschrift: Et sic est finis huius operis. Deo gracias. Zur Überlieferung s. R. Veit, Das Buch der Fieber des Isaac Israeli und seine Bedeutung im latei nischen Westen. Wiesbaden 2004. (176ra, 212vb) Verse aus Regimen Sanitatis Salernitanum (Ed.: S. Renzi, Collectio Salernitana V. Naples 1859). 30.1 (176ra) am unteren Blattrand vier Verse aus Regimen Sanitatis Salernitanum (Ed. [V. 1 und 2] Renzi V. 2924–2925). Tit.: Metra. – Inc.: Est triplex tipus rigor oripilacio frigus / … – Expl.: … A causa nigra sit orip pilacio vera / Est quartus tremor quod febres circuit has tres. Die letzten zwei Verse nicht bei Renzi. 30 30.2 (212vb) Sechs Verse aus Regimen Sanitatis Salernitanum (Ed. Renzi V. 1588–1589, 1604–1605, 1590–1591). Inc.: Grana quater quinque scrupuli pro pondere summe / … – Expl.: … / Exagium solido difert in nomine solo. 31 (212v) am unteren Blattrand Notiz. Inc.: Item nota quod febricitantibus acutatarum (!) febrium et etiam aliis qui paciuntur … – Expl.: … in lecto ut bene possit resistere et hoc per unam horam. 32 (213ra–218vb) Synonyma Rasis (Thorndike–Kibre 73, 81). Auch Serapion und Mesue zuge schrieben. Inc.: [Allia]seph puncti rubei qui sunt ut pulicis morsus et fiunt in omnibus membris … – Expl.: … [V]egi id est vitreolum. [V]erusca id est pondera etc. Schlussschrift: Expliciunt Synonima Rasis. 33 (218vc) Notizen. Tit.: Questio utrum spiritus sit subiectum sanitatis. – Inc.: Spiritus est vapor temperatus genitus ex parte subtiliori sanguinis … – Expl.: … dicitur de turbulencia oculi. Darunter: Venetus color id est celescis (!). (219r) Besitzvermerk (s. bei G). (219v) nur Rahmung. L. Š. Cod. 754 329 Cod. 754 THEOLOGISCHE SAMMELHANDSCHRIFT, u. a. IOHANNES GERSON Pap. 26 Bl. 280 × 195. Tirol, 3. Viertel 15. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Identische Was serzeichen: u. a. Innsbruck, ULB Tirol, Cod. 8 (dat. 1459), Cod. 101 (dat. 1464). Lagen: VI12 + V22 + 426. Bl. 27 als Spiegel auf dem HD aufgeklebt. In der Mitte der Lage 2 in Textura beidseitig beschriebener Pergamentfalz (zwei Spalten, Rubrizierung, u. a. Ps 26,14). Am Ende der Lage 1 Reklamante. Bis Bl. 6 am unteren Blattrand mittig zeitgenössische Foliierung. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S. Schriftraum 220 × 125/135, von Tintenlinien gerahmt, zu 36–40 Zeilen. Textualis cursiva mit variierender Schriftgröße von zwei Händen: 1) Bl. 1r–12v; 2) Bl. 13r–26v. A: Bl. 1r–12v schwarze Unterstreichungen, Paragraphzeichen. Aussparungen für zwei- und dreizeilige Lom barden. Vereinzelt rote Auszeichnungsstriche. Bl. 13r und 18r–26v Überschriften sowie zwei- und drei zeilige Lombarden, tw. mit Punktverdickungen und Ausläufern. Repräsentanten. Bl. 16v Nota-Hinweis mit Segenshand O vere nota. Ab Bl. 14r Oberlängen in der ersten bzw. Unterlängen in der letzten Zeile bisweilen cadellenartig gedehnt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 18. Jh. Buchblock tw. nur mehr schwach mit dem Einband verbunden, Rücken tw. freiliegend. Signaturschild (s. bei G). Schnitt rot gesprenkelt. Spiegel Pap. 18. Jh. Am Spiegel des VD Rahmung, Exlibris und Signaturen (s. bei G), am HD nur Rahmung. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol über geben. Am Spiegel des VD alte Signaturen der ULB Tirol II6 D […] (?, Tinte, radiert), II4 C 17 (Bleistift), 754 (Tinte). Am Rücken bzw. VD gelbes Signaturschild der ULB Tirol 754. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Wilhelm I 137. – Peintner, Stiftsbibliothek 121. – J. Mangei, Kartäuserorden und Visionsliteratur im Mittel alter und in der Frühen Neuzeit, in: S. Lorenz (Hrsg. in Zusammenarbeit mit dem Institut für Geschichtliche Landeskunde und Historische Hilfswissenschaften der Eberhard-Karls-Universität Tübingen und dem Instytut Informacji Naukowej i Studiów Bibliogicznych der Universität Warschau), Bücher, Bibliotheken und Schriftkultur der Kartäuser. Festgabe zum 65. Geburtstag von Edward Potkowski (Contubernium 59). Stuttgart 2002, 289–316, hier 301 und Anm. 76. – D. D. Martin, Carthusians as Advocates of Women Visionary Reformers, in: J. M. Luxford, Studies in Carthusian Monasticism in the Late Middle Ages. Turnhout 2009, 127–153, hier 139. – Katalog ULB Tirol VIII 269, 530. 1 (1r–11v) Iohannes Gerson: De oratione et suo valore (Ed.: Glorieux, Oeuvres II 169–191). 2 (11v–13r) ‹Dionysius Carthusianus›: Revelatio cuiusdam Carthusiensis in missa, dat. 1454 (vgl. Dionysius Carthusianus, Opera Omnia, Bd. 1, XLI–XLVIII). Bl. 11v Vermerk am Rand [V]isio cuiusdam Karthusiensis […]c persecucione seu statu sancte matris ecclesie anno domini 1454. Rubrica. Inc.: [Q]uidam religiosus ordinis Cartusiensis monasterii in Ruremunda in solempnitate purifica cionis gloriosissime virginis Marie in summa missa toto conventu presente et inspiciente ab exte rioribus sensibus … – Expl.: … Accidit autem hec visio currente videlicet anno domini milesimo quadringentesimo quinquagesimo quarto duravitque revelacio ista per integram circiter horam quia usque ad finem misse sedit in ea frater quasi immobilis et abstractus sicut nunc cecitatum est ad laudem et gloriam omnipotentis Dei qui est super omnia Deus gloriosus et adorandus in secula seculorum benedictus Amen. Zur handschriftlichen Überlieferung und zum Verhältnis der Textredaktionen bzw. -fassungen vgl. Mangei 289–316. 330 Cod. 754 3 (13r–18r) Epistola de visione cuiusdam fratris ordinis Carthusiensis in Ruremunda. Tit.: Sentimentum cuiusdam Cartusiensis super infestacione Thorcorum. – Inc.: Venerande ac predilecte pater ad satisfaciendum obediencie michi per paternitatem vestram nuper iniuncte sub actu visitacionis videlicet ut scribere vobis deberem quid senciam … – Expl.: … propter peccata nostra ut animabus saltem nostris provideat de salute eterna. Quod nobis prestare dignetur pater et filius et spiritus sanctus. Inc. u. a. Melk, Stiftsbibl., Cod. 1799, Bl. 78r. Lit.: Martin 127–153, bes. 139–142. 4 (18r–26v) ‹Ps.-Nicolaus de Dinkelsbühl›: Speculum artis bene moriendi (Druck: u. a. GW 2571. – R. Rudolf, Ars moriendi. Von der Kunst des heilsamen Lebens und Sterbens [Forschungen zur Volkskunde 39]. Köln, Graz 1957, 75. Madre 292. Bloomfield Nr. 1076). Gegenüber Druck gekürzt. Tit.: Tractatus de arte moriendi magistri Iohannis Gerson. – Inc.: Cum de presentis exilii [mise ria] mortis transitus propter moriendi impericiam … – Expl.: … ut prius antequam mors ipsum arripiat id ipsum scire mori discat. Quod nobis prestare dignetur pater et filius et spiritus sanctus Amen. Zur handschriftlichen Überlieferung, zum Werk und zur Verfasserfrage (in den Hs. u. a. auch Albertus Magnus, Dominicus de Capranica, Henricus de Langenstein, Iohannes Nider, Matthaeus de Cracovia, Thomas Peuntner zugeschrieben) s. Madre 292–295 und Rudolf 75–82. Bei Madre 292–295 unter den Opera dubia des Nicolaus de Dinkelsbühl angeführt, von Rudolf zu den echten Werken gezählt. Vgl. K. Schneider, „Speculum artis bene moriendi“. Verfasserlexikon2 IX 40–49. 5 (26v) Votivmessen und Gebete für Verstorbene (nicht im Druck für De arte moriendi nach weisbar). Inc.: Quidam egregius doctor in sermone publice predicavit quod quicumque devota intencione infrascriptas missas legeret seu legi disponeret pro anima … – Expl.: … quibus completis anima eidem apparuit leta et liberata magnas grates referens. Fünf Messen. Inc.: Prima missa est de exaltacione sancte crucis … In secunda missa sic domine Ihesu Christe qui iudicium innocentis sustinuisti … In tercia sic domine Ihesu Christe qui innocenter et indigne in cruce es conclamatus … In quarta missa sic domine Ihesu Christe per vulnera piissimi corporis et cordis tui … In quinta sic domine Ihesu Christe rex celorum per sepulturam tuam dignare animam … – Expl.: … ut ipsa a te numquam separetur in perpetuum. Parallelüberlieferung: Weimar, Herzogin-Anna-Amalia Bibl., Cod. Q 108, Bl. 316r. Schlussschrift: Amen dicat hec fidelis / qui regnare vult in celis. P. A. Cod. 755 331 Cod. 755 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 12 Bl. 270 × 200. Neustift oder Wiener Raum (?), 6. Jz. 15. Jh., 1481 (?) und 16. Jh. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasserzeichenbefund. Lage: VI12. Bl. 1r–9v am oberen Blattrand mittig in Rot Werktitel des jeweiligen Textabschnittes, auf Recto und Verso aufgeteilt, stark beschnitten. Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Bl. 4 neuzeitliches gelbes Papiersignakel der ULB Tirol. S: Schriftraum 200 × 140. Zwei Spalten, von Tintenlinien gerahmt, Bl. 1ra–9vb zu 42–46, Bl. 10ra–12vb zu 27–37 Zeilen. Zwei Hände: 1) Bl. 1ra–9vb Bastarda, möglicherweise dat. [14]81 (Bl. 3vb); 2) Bl. 10ra–12vb Humanistische Kursive 16. Jh. Nach Goy 424 möglicherweise in Neustift entstanden. A: Bl. 1ra–9vb rote Auszeichnungsstriche, Unterstreichungen und Überschriften. Bl. 1ra dreizeilige, sonst zwei zeilige rote Lombarden. Am oberen Blattrand Buchtitel in Rot, beschnitten, tw. zur Gänze w eggeschnitten. Bl. 10r–12v keine Rubrizierung. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Am Rücken bzw. VD Signaturschild der ULB Tirol. Schnitt rot gesprenkelt, kaum sichtbar. Spiegel Pap. 18. Jh., am Spiegel des VD Exlibris und Signaturen (s. bei G), Spiegel des HD leer. G: Hs. möglicherweise in Neustift entstanden; andernfalls Zeitpunkt und Art der Erwerbung unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II6 D […] (? Tinte, radiert), II4 C 18 (Bleistift) und 755 (Tinte). Am Rücken bzw. VD gelbes Signaturschild der ULB Tirol 755. Bl. 1r Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. L: Goy 424, Nr. 135. – Peintner, Stiftsbibliothek 122. – Katalog ULB Tirol VIII 269, 530. 1 (1ra–3vb) Iohannes Gerson: De probatione spirituum (Ed.: Glorieux, Oeuvres IX 177–185. – Bloomfield Nr. 4280). Tit.: Sequitur alius tractatulus specialis de probacione spirituum editus per supra nominatum arcium et sacre theologie professorem m. Io. Gerson [in] concilio Constanciensi. (3vb) unmittelbar an das Explicit anschließend Datierung (?) in Rot: LXXXI. 2 (4ra–8ra) Hugo de Sancto Victore: De orando Deum (PL 176, 977–988. – Goy 424, Nr. 135. Bloomfield Nr. 4865). Gegenüber PL tw. leicht abweichend. Ohne Prolog, Beginn der einzelnen Kapitel in der Hs. nicht hervorgehoben. Tit.: Hugo de Sancto Victore de orando Deum etc. – Inc.: Quo studio et quo affectu a nobis oran dus sit Deus … – Expl.: … et gratissimum Deo sacrificium in ara crucis adoletur ibi Amen etc. Bl. 6v nur Rahmung, Forts. des Textes von Bl. 6vb auf Bl. 7ra ohne Textverlust. 3 (8rb–vb) Iohannes Gerson: De modo confessionis et absolutionis sacramentalis (Ed.: Glorieux, Oeuvres IX 646–648). 4 (8vb–9va) Iohannes Gerson: De modo excommunicationum (Ed.: Glorieux, Oeuvres IX 92–94). 5 (9va–b) Direkt anschließend ohne Hervorhebung: ‹Iohannes Gerson (?)›: De iniunctione paeni tentiae (Ed.: Glorieux, Oeuvres I 48. – Bloomfield Nr. 5390). 6 (10ra–12vb) Auszüge aus verschiedenen lat. Autoren. (10ra–11rb) Publius Terentius Afer, mit vereinzelten Auszügen aus dem Kommentar des I ohannes Calphurnius und des Aelius Donatus (Druck: Lipsiae 1830. Ed.: R. Kauer, W. M. Lindsay. Oxonii 1958. Kommentar nur im Druck enthalten). Vereinzelt Einschübe aus Werken anderer Autoren. 332 Cod. 755 (10ra) Tit.: Andria: Prolog, 22–23. – 1,1,60–61. – 1,1,68. – 1,1,77–79. – 1,2,191. – 1,3,218. – 1,5,266. – 2,1,305–306. – 2,1,309. – 2,1,330–331. – 2,5,425–427. – 3,3,555. – 5,3,903. – 5,4,940. – 5,5,959–961. (10ra–b) Tit.: Eunuchus: Prolog, 41. – 1,2,178. – 2,1,225–226. – 2,2,248–249. – 2,2,251–253. – 2,2,265. – 2,2,276. – Kommentar zu 2,3,379. – 2,3,381. – 3,1,408–409. – 3,1,426. – 3,1,445. – 3,2,476. – 3,2,490–491. – 3,2,494–495. – 4,2,640–641. – 4,4,722. – 4,4,732. – 4,6,761–762. – 4,7,788–789. – Plautus: Miles gloriosus 1,1,68. – Forts. Terenz 5,1,832. (10rb–vb) Tit.: Heautontimorumenon. Cicero: De officiis 1,22. – 1,2,177. – 1,2,202. – 2,2,239– 240. – 2,3,256. – 2,3,314. – 2,3,364. – Kommentar zu 2,4,382. – 3,3,419. – Cicero: Cato maior de senectute 3,7. – Forts. Terenz 1,1,77. – 1,2,209–210 – 3,3,481–483. – 3,3,503–506. – 3,3,573. – 3,3,566–567 – 3,3,578. – 3,3,611. – 4,1,646. – 4,1,648. – 4,1,666. – 6,2,675. – Kommentar zu 4,3,687. – Kommentar zu 4,3,693. – 4,3,706. – Kommentar zu 4,4,732. – Kommentar zu 4,5,753. – 4,5,796. – 4,6,805. – 4,7,839. – 5,1,887. – 5,1,922–923. – 5,2,962–963. – 5,3,991–993. (10vb–11ra) Tit.: Adelphis: 1,1,74–75. – 1,2,98–99. – 1,2,105. – 2,2,216. – 2,3,254. – 3,3,386– 388. – 3,3,414–416. – 3,3,399. – Pii esse debemus in parentes in saguine (!) nobis iniunctos ad neccesitatem precipue autem post debemus esse in Deo. – 3,4,508–509. – 4,1,537. – 4,7,734. – 4,7,739–741. – 5,3,833–834. – 5,4,855–856. – 5,4,861. (11ra–b) Tit.: Phormio: 1,1,41–42. – 1,1,55–56. – 1,2,77–78. – 1,2,138. – 1,3,172. – 1,4,203. – 2,1,264–265. – 2,1,275–277. – 2,2,330–331. – 2,4,454–456. – 3,2,487. – 4,1,587. – 4,5,726. – 5,1,756–758. – 5,2,766–767. – 5,8,903–904. – 5,8,948–950. (11rb) Tit.: Echira (recte: Hecyra): 2,1,199–200. – 2,1,202–204. – 4,4,660–661. – Kommentar zu 4,4,687. (11va–b) Quintus Horatius Flaccus: Carmina (Ed.: D. R. Shackleton Bailey, Q. Horati Flacci Opera. Berolini et Novi Eboraci 2008). Tit. stark beschnitten: Horacii par[…]. Ode in libro primo. (11va) 1,3,37–40. – 1,4,13–20. – 1,10,7–9. – 1,35,26–28. (11va) Decimus Iunius Iuvenalis: Saturae (Ed. J. A. Willis. Stutgardiae 1997) 9,128–129. (11va–b) Quintus Horatius Flaccus: Carmina (Ed. wie oben) 2,2,9–12. – 2,3,21–28. – 2,10,9– 12. – 2,10,19–20. – 2,11,16–18. (11vb) Publius Ovidius Naso: Ars amatoria (Ed. E. J. Kenney. Oxonii ²1995) 2,279–280. (11vb) Gaius Sallustius Crispus: De coniuratione Catilinae (Ed. L. D. Reynolds. Oxonii 1991) 52,29. (11vb) Quintus Horatius Flaccus: Carmina (Ed. wie oben) 2,13,13–14. (11vb–12ra) Quintus Horatius Flaccus: Carmina (Ed. wie oben) 2,14,1–4. – 2,16,13–16. – 2,16,25–28. – 2,18,15–16. – 3,1,13–15. – 3,1,16–20. – 3,2,13. – 3,2,29–32. – 3,2,21–22. – 3,3,9– 10. – 3,29,29–32. (12ra) Decimus Iunius Iuvenalis: Saturae (Ed. wie oben) 2,3. – 13,209–210. (12ra) Quintus Horatius Flaccus: Sermones (Ed. wie oben) 2,8,61–63. (12ra) Iuvenalis: Saturae (Ed. wie oben) 6,53–54. (12ra) Quintus Horatius Flaccus: Carmina (Ed. wie oben) 4,7,7–8. – 4,7,14–18. (12rb) Quintus Horatius Flaccus: Ars poetica (Ed. wie oben) 158–174. – 25–26. – 139. – 359. – 337. – 343. – 350. – 356. – 390. – 385. – 365. – 476. (12va) Quintus Horatius Flaccus: Sermones (Ed. wie oben) 1,1,106–107. – 1,2,86–91. – 1,3,68– 69. – 2,3,325–326. – 2,3,94–99. – 2,5,7. – 2,6,94–97. (12va–b) Quintus Horatius Flaccus: Epistolae (Ed. wie oben) 1,1,45–46. – 1,2,14. – 1,2,27–31. – 1,2,57–59. – 1,2,69–70. – 1,4,14. – 1,5,12. – 1,5,16–20. – 1,6,27–28. – 1,6,36–38. – 1,7,98. – 1,8,11. – 1,12,4. – 1,10,47–48. – 1,16,48. – 1,16,52–53. – 1,16,78–79. A. P. Cod. 756 333 Cod. 756 MATTHIAS ENGELSCHALK Pap. 53 Bl. 275 × 200. Süddeutscher Raum, 1479. B: Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Lagen: 4.VI48 + 553. Bl. 49–53 eingehängte Einzelbl. In den Lagenmitten beidseitig beschriebene Pergament fälze, tw. rubriziert (Makulatur aus liturgischen Hs.). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. S: Schriftraum 255 × 165. Zwei Spalten, von Tintenlinien gerahmt, zu vorwiegend 43 bzw. 46 Zeilen. Bastarda, dat. 1479 (Bl. 53ra). A: Rote Auszeichnungsstriche, Unterstreichungen, zweizeilige (Bl. 1ra vierzeilige) rote Lombarden mit Punkt verdickungen. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Rücken: Pap. oben eingerissen und geklebt, drei einfache Bünde. Im zweiten Feld Signaturschild (s. bei G). Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Signaturen und Stempel (s. bei G), Spiegel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neustift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] (radiert, Tinte), II4 C 19 (Bleistift) und 756 (Tinte). Am Rücken gelbes Signaturschild der ULB Tirol 756. Bl. 1r Stempel der ULB Tirol. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Am Spiegel des VD Stempel der Soprintendenza Tridentina 20. Jh. L: Stegmüller RB Nr. 5550 Suppl. – Stefani 118. – Peintner, Stiftsbibliothek 121. – Katalog ULB Tirol VIII 269, 530. ‹Matthias Engelschalk›: De passione Christi secundum quattuor evangelistas (Stegmüller RB Nr. 5550 Suppl., mit Nennung der Hs.) Tit.: Passio domini nostri Ihesu Christi secundum Matheum Marcum Lucam et Iohannem. – Inc.: In illo tempore dixit Ihesus discipulis suis. Scitis quia post biduum pascha fiet et filius hominis tradetur ut crucifigatur … (Mt 26,2–25). Quia nullus ewangelistarum materiam dominice passionis plene ac complete conscripsit … – Expl.: … vindicta inflicta Iudeis propter innocentem Christi mortem. (53ra) Datierung (in Rot): Anno MoCCCCo et LXX9. Darunter: Et sic est finis. Darunter von späterer Hand (in Rot) Carnem. (53rb) Nachtrag: 17. Jh. (kursiv): Commonefacio vos enim fratres mei collegaeque devoti, futuri festi nativitatis Christi, ad sacras ordines nempe ipsasque assumere promoveritis vosque ideo summa doctrinae providentia provideatis ut digne eas percipere possitis. Darunter wiederholt (leicht abgeändert) in Rot. (53v) leer. C. S. 334 Cod. 776 Cod. 776 MUSIKALISCH-ASTRONOMISCHE SAMMELHANDSCHRIFT Pap. I, 71, I Bl. 210 × 155. Neustift (?), Anfang 16. Jh. (u. a. 1502). B: Drei Teile: I (I, 1–22), II (23–46), III (47–71, I). Buchblock nachträglich leicht beschnitten (vgl. Rand notizen). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol. Bl. 22 und 47 neuzeitliche gelbe Papiersignakel der ULB Tirol. E: Bibliothekseinband: Perg. über Pappe. Italien, 20. Jh. Neubindung im Zuge der Restaurierung (kein Restaurierbericht vorhanden). Kapitale mit weißem und grünem Zwirn umstochen. Schnitt rot gesprenkelt. Spiegel Pap. 20. Jh., leer. G: Hs. möglicherweise in Neustift entstanden und urspr. mit Innsbruck, ULB Tirol, Cod. 1015 in einer Hs. vereinigt (vgl. die übereinstimmenden Wasserzeichen sowie die Größe der Hs.). Gegebenenfalls wohl im Zuge der Neuaufstellung der Neustifter Bibliothek in der 2. H. 18. Jh. (wie dies auch bei anderen Hs. und In kunabeln gemacht wurde) in Einzelhandschriften aufgelöst. Vor Bl. 1 Exlibris der Stiftsbibl. Neustift 18. Jh. eingebunden, urspr. wohl am Spiegel des VD (nicht mehr vorhanden) aufgeklebt. 1809 anlässlich der vor übergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Bl. 1r alte Signatur der ULB Tirol 776 und Stempel der ULB Tirol 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italie nischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Tridentina 20. Jh. und der Stiftsbibl. Neustift 20. Jh. Vor Exlibris zusätzlicher schmaler Papierstreifen eingebunden ms. 776. Biblioteca dell’abbazia di Novacella – Varna-Bressanone (Bolzano). L: Wilhelm I 137. – Kristeller I 439. – P. Fischer, The Theory of Music from the Carolingian Era up to 1400 II: Italy. München, Duisburg 1968, 73. – Neuhauser, Neustift 87. – Peintner, Stiftsbibliothek 126. – M. H. Peintner, Psallite Deo! Zur Musikgeschichte des Stiftes Neustift, in: 850 Jahre Augustiner Chorherrenstift Neustift 134–153, hier 137 (ohne Nennung der Sign.). – M. Peintner, Musikgeschichte des Stiftes, in: 850 Jahre Neustift. Katalog 94–100, hier 97 (ohne Nennung der Sign.). – Stefani 119. – Neuhauser, Musik geschichtliche Quellen 194. – M. Bernhard, Zur Rezeption der musiktheoretischen Werke des Hermannus Contractus, in: W. Pass, A. Rausch (Hrsg.), Beiträge zur Musik, Musiktheorie und Liturgie der Abtei Rei chenau (Musica mediaevalis Europae occidentalis 8). Tutzing 2001, 99–126, hier 115–116, 120. – Katalog ULB Tirol VIII 426, 530. I (I, 1–22) B: B. 1r leicht, Bl. 22v stark verschmutzt und fleckig. Bilder und Nachweise der Wasserzeichen s. WZMA. Wasserzeichenbefund mit Datierung übereinstimmend. Lagen: (I–1)I (Vorsatzbl. 20. Jh.) + VI12 + V22. Gegenbl. zum Vorsatzbl. als Spiegel auf dem VD aufgeklebt. S: Schriftraum stark wechselnd, seitlich von Blindlinien begrenzt, Zeilenzahl wechselnd. Flüchtige Humanis tische Kursive, dat. 2. Jänner 1502 (Bl. 12v). Überschriften bzw. Textanfänge sowie einzelne Wörter im Text in Auszeichnungsschrift. Zahlreiche Notenbeispiele tw. in gotischer Choralnotation mit c- und f-Schlüssel auf Vier- bzw. Fünfliniensystemen, tw. in Quadratnotation auf Drei- bis Fünfliniensystemen. Zahlreiche geschwungene Klammern als strukturierende Gliederungshilfen. A: Rote Unterstreichungen. Bl. 1v und 13r drei- bzw. zweizeilige blassrote Lombarden mit Punktverdickungen. Bl. 8v und 9r Schemata zu Tonschlüssen (Primus et secundus toni finiuntur in D sol re … septimus et octavus finiuntur in G sol re ut [gravi naturaliter sive regulariter]). 1 (1r–12v) Musicae modulationis commendatio. (1r) Signatur und Stempel (s. bei G). Tit.: Musica gregoriana. (1v) Tit.: Musice modulacionis commendacio. – Inc.: Sicut apotheca est salubris ita musica delectabilis quia dulcedine sua … – Expl.: … Artem discamus usum vero teneamus. (8v–9r) Schemata zu Tonschlüssen. (12v) Schlussschrift mit Datierung: Finis adest fauste anno 1502 die IIa Ianuarii (2. Jänner 1502). Cod. 776 2 335 (13r–20v) Musiktraktat. Inc.: Musica humane vite necessaria nedum apud Grecos aut Romanos verum etc. apud barbaros semper magna laude prosecuta est … – Expl.: … Excedes duplum – Non caudata – Non quadrata – Ascendit tunc quelibet videlicet brevem / Descendit tunc prima videlicet longam et quevis sequens brevem. Enthält zwölf Kapitel. (21r–22v) leer. II (23–46) B: Pap. tw. fleckig, Bl. 23r–24r stark verschmutzt. Bilder und Nachweise der Wasserzeichen s. WZMA. Identi sche Wasserzeichen: u. a. Innsbruck, ULB Tirol, Cod. 1015 (dat. 1509). Lagen: 2.VI46. S: Schriftraum stark wechselnd, seitlich von Blindlinien begrenzt, stark wechselnde Zeilenzahl. Zwei Hände: 1) Bl. 23r–29v flüchtige Humanistische Kursive, wohl von derselben Hand wie T. I (dat. 1502); 2) Bl. 31r– 46v regelmäßige Bastarda, Überschriften bzw. Textanfänge und einzelne Wörter im Text jeweils in Aus zeichnungsschrift. Zahlreiche Notenbeispiele in gotischer Choralnotation mit c- und f-Schlüssel auf Vierund Fünfliniensystemen. Geschwungene Klammern als strukturierende Gliederungshilfen. 3 (23r–46v) Tractatus de divisionibus in musica. Tit.: [M]usica Isidoro diffiniente est recte canendi sciencia ad laudem Dei finaliter adinventa. – Inc.: Et dedumur (!) a Musis que per Bocacium XI libro De genologia (!) Deorum gentilium … – Expl.: … Examen clavium. Hec habui amantissimi scholares que vobis et condiscendam musicam practicam et planam digna ducerim colligenda que et si meo ingenio minime invenerim sed maiorum nostrorum tradicionibus et informacione perfecto magistri mei in his rebus Udalrici Flordigalis per benigna dedicerim (!). Isto tamen ordine quem vobis omnibus (gestrichen: meis) magis conveniente existimavissem in unum opusculum collegi quare vos omnes oro obtestor laborem hunc meum non aspernemini unde vobis non modicum utilitatis ubi non vos ipsos in hiis meis sudoribus exercitare non pugnerit (?) sit accessurum. Lit.: Bernhard 115–116, bezogen auf Bl. 27v Ter terni sunt modi … III (47–71, I) B: Pap. bes. Bl. 47r, 61r und 71v fleckig, Bl. 47v–50r Tinte stark verwischt (Tintenfraß?). Bilder der Wasserzei chen s. WZMA. Lagen: VII60 + (VI–1)71 + (I–1)I* (Nachsatzbl., 20. Jh.). Gegenbl. zum Nachsatzbl. als Spie gel auf dem HD aufgeklebt. Gegenbl. zu Bl. 61 herausgeschnitten (ohne Textverlust), Gegenfalz auf Bl. 71 aufgeklebt. S: Schriftraum 150/160 × 95/100. Seitlich von Blind-, Bl. 55r–60r von Tintenlinien begrenzt, zu 29–33 Zeilen. Bastarda mit wechselnder Tinte. Oberlängen in der ersten Zeile bisweilen gedehnt. Überschriften bzw. Text anfänge in Auszeichnungsschrift. Ergänzungen am Blattrand wohl von derselben Hand. A: Bl. 68v rote Auszeichnungsstriche. Zeigehände. Bl. 61r Platz für siebenzeilige Initiale freigelassen, nicht ausgeführt. Bl. 68r Fixsterntabelle mit Spalten- und Zeilenbezeichnungen in Rot. Bl. 71v Skizze einer Alhi dade (Vorrichtung zur Winkelanzeige oder Winkeleinstellung am Astrolabium). 4 4.1 (47r–71v) Astronomische Texte. (47r–60v) ‹Robertus Anglicus (?)›: De astrolabio (Druck: GW M38333. – Thorndike–Kibre 331,4 und 1228,5). Inc.: Quia plurimi obnimiam quandoque accurationem et magnam scriptorum sentenciam canones utilis astrolabii declarantes intelligere … – Expl.: … pars altitudinis rei tui adde. Enthält 53 Canones. Parallelüberlieferung: u. a. Melk, Stiftsbibl., Cod. 601, Bl. 18ra. Lit.: F. J. Carmody, Arabic astronomical and astrological sciences in latin translation. A critical bibliography. Berkeley, Los Angeles 1956, 19 Nr. 14; G. Rosinska, Instrumenty astronomiczne 336 Cod. 780 na Uniwersytecie Krakowskim w 15 wieku (Studia Copernicana 11). Wroclaw u. a. 1974, 105– 106, Nr. 12; Dies., Scientific writings and astronomical tables in Cracow I: Initien (Studia Coper nicana 22). Wroclaw u. a. 1984, Nr. 1779. 4.2 (61r–71v) ‹Prosdocimus de Beldemandis›: Compositio astrolabii (Druck: GW M38333. – Thorn dike–Kibre 1164,1). Inc.: [Q]uamvis de astrolabii composicione tam modernorum quam veterum dicta habeantur … – Expl.: … restringens in foramine illud inmitte ut teneat. Et habebis astrolabium perfectum. Parallelüberlieferung: Rostock, UB, Mss. math.-phys. 1, Bl. 173v; Melk, Stiftsbibl., Cod. 794, Bl. 369r. Lit.: P. Spunar, Repertorium auctorum Bohemorum provectum idearum post universitatem Pra gensem conditam illustrans I (Studia Copernicana 25). Wratislaviae u. a. 1985, 126, Nr. 334 (unter den Werken des Cristannus de Prachaticz = de Praga); N. Severino, Il libro degli astrolabi. La storia dell’Astrolabio da Tolomeo a Regiomontano. Roccasecca 1994, 39; G. Rosinska, Ins trumenty astronomiczne (s. o.) Nr. 11; Dies., Scientific writings (s. o.) Nr. 1715; A. Hadravová, P. Hadrava, Křišťan z Prachatic. Stavba a užití astrolábu. Praha 2001. 4.3 (71v) Notiz zur Verwendung der Alhidade mit Skizze (vgl. R. B. Thomson, Pseudo-Mash’allah. On the Astrolabe part II. Toronto 2012, 68). Inc.: Si per allidadam horariam vis horam diei invenire naturalem poni aldidam (!) super al titudinem medie diei illius in dorso astrolabii suspensi … – Expl.: … in divisionibus erit hora quesit[a]. (Ir–v) leer. A. P. / U. S. Cod. 780 GAIUS SALLUSTIUS CRISPUS Pap. 86 Bl. 210 × 145. Oberitalien (?), Ende 15. Jh. B: Bl. 10–15 und 46–49 beinahe, Bl. 72/73 vollständig lose. Buchblock nachträglich beschnitten (vgl. Rand bemerkungen mit Textverlust). Bilder und Nachweise der Wasserzeichen s. WZMA. Datierung nach Wasser zeichenbefund. Lagen: XII24 + VI36 + XII59(60) + XIII85(86). Am oberen Blattrand Foliierung 19. Jh. durch die ULB Tirol; Fehler in der Zählung: 41 doppelt angeführt. Reste von zwei neuzeitlichen gelben Papiersigna keln der ULB Tirol. S: Schriftraum Bl. 1r–v: 160/175 × 90/105, Bl. 2r–35v: 170 × 70/75, Bl. 37r–85v: 165/180 × 80/85, seitlich von Blindlinien begrenzt, Bl. 1r–v zu 35 Zeilen, Bl. 2r–35v zu 27–28 Zeilen, Bl. 37r–85v zu 28–35 Zeilen. Humanistische Kursive, wohl von einer Hand mit unterschiedlichem Duktus. Notizen von zeitgenössischer Hand. A: Randmarkierungen in Form von vereinfachtem Perlenbesatz. Bl. 40r Zeigehand. Bl. 2r–28r zweizeilige tintenfarbene Lombarden mit Punktverdickungen, anschließend Platz für Lombarden freigelassen, nicht aus geführt. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. Neustift, 2. H. 18. Jh. Pap. insbes. am HD und am Rücken beschädigt. Am Rücken Signaturschild der ULB Tirol. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris und Signaturen (s. bei G) sowie Bleistiftnotiz (s. bei Inhalt), Spie gel des HD leer. G: Zeitpunkt und Art der Erwerbung durch Neustift unbekannt. Am Spiegel des VD Exlibris der Stiftsbibl. Neu stift 18. Jh. 1809 anlässlich der vorübergehenden Aufhebung des Klosters Neustift der ULB Tirol übergeben. Am Spiegel des VD alte Signaturen der ULB Tirol II […] B 11 (Tinte, radiert), II4 D 9. S. 2 (Bleistift) und 780/(I) (Tinte, Bleistift). Am Rücken gelbes Signaturschild der ULB Tirol 780. Bl. 1r Stempel der ULB Tirol Cod. 780 337 19. Jh. 1921 aufgrund des Friedensvertrages von St. Germain an den italienischen Staat abgegeben und von diesem 1929 dem Kloster Neustift als Dauerleihgabe überlassen. Bl. 1r Stempel der Soprintendenza Triden tina 20. Jh. L: Peintner, Stiftsbibliothek 126 (hier fälschlich als Cod. 789 bezeichnet). – Neuhauser, Neustift 84. – Katalog ULB Tirol VIII 434, 530. (VDS) Bleistiftnotiz 19. Jh.: NB. Continet etiam librum de bello Iugurthino; hoc est in catalogo notandum. 1 (1r–v) Iulius Pomponius Laetus: Begleitbrief an Augustinus Maphaeus, der Sallustedition voran gestellt (GW M39655). Tit.: Augustino Maffeo rerum Ro. thesauro Pomponius Letus. – Inc.: M. Valerius Probus unice vetustatis amator … – Expl.: … ab omni offensiuncula vindicabis. Vale foelicissime. 2 (2r–36v) Gaius Sallustius Crispus: De coniuratione Catilinae (Ed. L. D. Reynolds. Oxonii 1991, 5–53). Mit zahlreichen Rand- und Interlinearglossen, u. a. Zitate aus Ovid (z. B. Bl. 2r Metamor phoses, Lib. 1, V. 84–86), Vergil (z. B. Bl. 2v Georgica, Lib. 2, V. 532–540), Iuvenal (z. B. Bl. 2v Saturae XV, V. 168). Tit.: C. Crispii Sallustii liber de coniuratione L. Sergii Catilinę. (36r–v) nur Rahmung. 3 (37r–85v) Gaius Sallustius Crispus: De bello Iugurthino (Ed. L. D. Reynolds. Oxonii 1991, 54–149). Mit zahlreichen Rand- und Interlinearglossen. Tit.: C. Crispi Sallusti liber de bello Iugurthino. P. K. 338 Sign. 14820 Sign. 14820 RITUALE NEOCELLENSE Perg. X, 85, II* Bl. 300 × 230. Neustift, Anfang 16. Jh. (mit Zusätzen, u. a. 1578). B: Dickes Perg. Lagen: 2II (Pap.) + (IV–2)VIII (Perg.) + 2X (Pap.) + (II+1)5 + 9.III59 + 2.(III–1)69 + 2.IV86(85) + III* (Pap.). Gegenbl. zu Bl. III und IV herausgeschnitten (ohne Textverlust), Bl. IX und X (ein Doppelbl.) auf Fälze aufgeklebt. Bl. IX und X urspr. kleineren Formates (300 × 210), Bl. IX am Blattrand durch zusätzli chen aufgeklebten Rahmen auf Größe des Buchblocks ergänzt, Bl. X auf anderes Papierblatt aufgeklebt. Bl. 5 eingehängtes Einzelbl. Gegenbl. zu Bl. 62 und 67 herausgeschnitten (ohne Textverlust). Am oberen Blattrand mittig zeitgenössische Foliierung in roten römischen Zahlen von I–LXXXV; Fehler in der Zählung: LXXII ausgelassen. Bl. 56v, 57rv, 58r, 62v, 63rv, 77rv Reste von braunen Ledersignakeln. S: Bl. 1r–85v: Schriftraum 225 × 155, von feinen Tinten- bzw. Blindlinien gerahmt, zu einheitlich 24 Zeilen auf feiner Tinten- bzw. Blindlinierung (mit Hilfslinien für Minuskeln). Zirkelstiche. Textualis formata (Textura mit Rotundaeinflüssen, verzierte Doppelschrägstriche als Skriptoriumseigenart?). Schreiber (?): F. H. (Bl. 85v). Bl. 64v und 67r kurze deutschsprachige Einschübe, Buchstaben nach rechts geneigt. Am Blattrand sehr vereinzelt Ergänzungen von späterer Hand. Bl. 85v Zusätze von zwei späteren Händen. Datierung 1578 (Bl. 72v) wohl nicht auf Haupthand bezogen, sondern auf spätere Nachträge, möglicherweise auf die Bl. 67v–71v nachgetragenen Seitenverweise (vgl. Register). Spätere Zusätze von verschiedenen Händen: Bl. IIIv: Schriftraum 240 × 150. Zwei Textspalten sowie zwei zusätzliche Spalten für die Blattangabe, seitlich jeweils von Blindlinien begrenzt, zu 34 Zeilen. Textualis formata (Textura) 16. Jh. Bl. IVr–VIIIr: Schriftraum 225/230 × 150/155. Von Blindlinien gerahmt, zu 23 Zeilen auf Blindlinierung. Antiquakursive wohl 17. Jh. Bl. IXr–Xr: Schriftraum 250 × 160. Zwei Spalten, von jeweils doppelten Tintenlinien gerahmt, zu ca. 30 Zeilen auf Blindlinierung. Antiqua 17./18. Jh. Bl. 86r: Schriftraum 235 × 165. Von Bleistiftlinien gerahmt, 21 Zeilen auf Bleistiftlinierung, diese jedoch nicht immer berücksichtigend. Vier verschiedene Hände. Antiqua bzw. Textura 17./18. Jh. N: Bl. 7v–9v, 19r–20v, 38v–42v, 65r: Gotische Choralnotation mit f- bzw. c-Schlüssel auf vier roten Linien. Kustoden. Reperkussionston mit einem ornamentalen Haarstrich. Rote Gliederungsstriche. A: Rote Auszeichnungsstriche, Überschriften, Verweiszeichen, Durchstreichungen und Zeilenfüllsel; wenige große Paragraphzeichen in (Dunkel-)Rot und auch Grün; (dunkel-)rote Benediktionskreuze (zumeist in Tatzenkreuzform, Bl. 17r–18r in den untersten Zeilen auch als lat. Kreuze mit Dekor). Zur Wasserweihe drei Zeichen (jeweils ca. eine Notenzeile hoch), die die schriftlichen Handlungsanweisungen visuell unterstützen (vgl. Cod. 194): Bl. 40v Kreuz (dunkelrot, mit verzierten Enden) zu hic insuffla per modum crucis sancte, Bl. 41v Stern (blau, mit vier spindelförmigen Strahlen) zu hic proiciatur aqua in quatuor partes und Bl. 43r Lilie (dreiblättrig mit Herzblatt als Gegenmotiv, blau mit roten Konturlinien) zu suffla ter in fontem in hunc modum. Vereinzelt aufwändigere (schwarz-)rote Cadellen in den obersten Zeilen (z. B. Bl. 73r); einige (cadellenartig) auf den unteren Seitenrand verlängerte Textualis-Buchstaben. In den Binnenfeldern der Tex tualis-Majuskeln bisweilen feine Doppelschrägstriche, tw. mit kleinen angesetzten Bögen (z. B. Bl. 51v, 10. Z. von unten). Im Register ein- und zweizeilige rote Initialbuchstaben, Bl. VIIv mit einfachem Dekor. Vielzahl von ein- bis zehnzeiligen Lombarden in Blau, in charakteristischem zu Türkis tendierendem (Hell-) Grün sowie in mehreren Rot- und Violetttönen, davon ca. 250 mit Fleuronnée verziert. Lombarden aller Farben können mit (wein-)rotem Fleuronnée kombiniert werden, blaue und rottonige Lombarden mit (oliv-) grünem Fleuronnée, hellrote und -grüne Lombarden mit (blass-)braunem und nur hellrote Lombarden mit (blass-)blauem Fleuronnée; häufig Punktverdickungen; v. a. in den untersten Zeilen einige aufwändigere Lombarden, deren Ausläufer in Achterschlaufen gelegt sind, z. B. Bl. 82v6 (sehr ähnlich in Cod. 100, vgl. z. B. Bl. 5va; aber wohl nicht von einer Hand); größere Lombarden (fünfzeilig und höher) fallweise kopf stempelartig sowie in diversen geometrischen Formen rot-blau oder rot-grün gespalten und mit zweifarbi gem Fleuronnée geschmückt (zu Bl. 49r, mit ausgesparten Blättchenranken, s. u.). In den Binnenfeldern vielfältige Formationen aus aneinandergereihten, länglichen, (leicht) blasigen K nospen (in verschiedenen Größen); gerne auch ondulierende, fallweise aus vollfarbigem Grund ausgesparte Ranken mit dreizackigen Blättern, deren Mitteladern tropfenförmig verdickt und mit kleinen Punkten oder Kreisen gefüllt sein können („Erbsenschotenmotiv“), z. B. Bl. 47r3, 76r3; selten geometrisch strukturierte Binnen felder, z. B. Bl. 63r Rautengitter mit eingeschriebenen Blüten. Konturlinie(n); als Besatz sehr oft Zweieroder Dreierkombinationen von Knospen bis hin zu Knospenreihen (jeweils im Wechsel mit Dornen), deren Endperlen in mehr oder weniger geschwungene Fäden auslaufen können; häufig kammartig abstehende, spitz zulaufende, tw. gebündelte Fadenausläufer (mit hakenartig geschwungenen Enden), die meist ortho gonal oder radial abstehen, aber auch – dies ist eher ungewöhnlich – an einer Initiale alle senkrecht nach oben ausgerichtet sein können (s. Bl. 75v1); seltener Fleuronnée-Leisten: aus mehreren Konturlinien und Sign. 14820 339 Knospenreihen aufgebaut, am Schriftspiegel entlang geführt, mit bes. stark geschwungen bzw. spiralig ein gedrehten Fadenausläuferenden (z. B. Bl. 24v: Leiste verknüpft die beiden Initialen). Bei größeren Fleuron née-Initialen Ausbildung von mehrlinig gerahmten Initialfeldern (z. B. Bl. 3r1) und Vollfarbigkeit möglich, s. türkisgrün ausgemaltes Binnenfeld Bl. 48r2 (s. auch bei figürlichem Schmuck) sowie die Initialen Bl. 12r2, 50v1, 52r4, 53r2, 73r2, bei denen die Buchstabenkörper (in Rot, Grün und nachgedunkeltem, jetzt ockerfar benem bis bräunlichem Gelb) tw. mit Blattranken (s. o. Binnenfeldranken) verziert, mit ausgemalten Profil rahmen umgeben und von einem mit (Blatt-)Ranken überzogenen, vollfarbigen Initialfeld hinterlegt sind, wobei die verwendeten Farben (z. B. Bl. 52r4 Violett für das Initialfeld) dünn aufgetragen wurden; Bl. 52r4 und 53r2 ohne Besatzfleuronnée. Figürlicher Dekor in (zumeist weinroter) Federzeichnung. Besatz: kleinere Blattmasken und Gesichter (Bl. 1v2, 4r1); diverse Profilköpfe mit langen Nasen (charakteristisch der gewellte Nasenrücken, z. B. Bl. 24r); Bl. 74r größere Profilbüste mit herausgestreckter Zunge. Binnenfeld: Bl. 29v Gesicht, Bl. 48r2 (Fronleichnam) goldener Kelch mit Hostie darüber (vollfarbig), Bl. 60v Mönchsbüste. Bl. 4v (die untere Seitenhälfte ausfüllend): Neustifter Wappen mit kartuschenartigem Schild, Geviert, Feld 1 und 4: auf rotem Grund je vier silberne Spitzen, Feld 2: Tau-Kreuz, Feld 3: Patriarchenkreuz auf Dreiberg; als Zimier Engel (in Wams mit weiten Ärmeln, auf dem Kopf florales Gebinde), dem zwei große spiegelsymmetrisch ange legte Heckenrosenvoluten (mit distelartigen Knospen) entwachsen, nach denen er die Arme ausbreitet. Der allergrößte Teil des verschwenderisch eingesetzten, variationsreichen Fleuronnée, das das Gesamt erscheinungsbild der Handschrift entscheidend prägt, stammt von der am Anfang des 16. Jh. nachweisbaren Hand, die auch in Cod. 405 (dat. 1501), Cod. 194 (dat. 1507; zu dieser Hs. zudem Textbezüge) und Cod. 142, T. I (dat. 1521) tätig war, und in beiden Bänden des Zollner-Graduales einige Initialen mit Fleuronnée geschmückt hat (s. jeweils Ergänzungshand 2, speziell die große Ähnlichkeit der zweifarbig gespaltenen Initialen mit ausgesparten Blättchenranken in Sign. 14820, Bl. 49r, und im ersten Gradualband, Bl. 187bisv). Die Formenvielfalt kann die Entscheidung erschweren, ob eine Variante vorliegt oder ob weitere Hände am Werk waren, so Bl. 65r (Notation) zwei Cadellen mit Profilköpfen, Bl. 85v (Nachtrag) Initiale mit Profil kopf, der von anderer Hand gezeichnet sein könnte (Besatz dem Hauptfleuronnée allerdings sehr ähnlich); Bl. 7r1, 18r, 21v1, 38r2 jeweils das O von Omnipotens sempiterne Deus mit ansonsten nicht verwendeten Überschneidungen bzw. Doppelungen im Buchstabenkörper und im Liniennetzdekor sowie plastisch wir kenden Fibrillen, wohl von anderer Hand, die wahrscheinlich auch Bl. 77v1 das d (mit Fleuronnée im Binnen feld) über ein hellrotes I gesetzt hat (vgl. weinrote Tinte). Der Aufbau der Binnenfeld-Blattranken erinnert an Deckfarbenranken, was vermuten lässt, dass der Florator auch als Deckfarbenmaler tätig war (oder zumindest Vorzeichnungen für ornamentalen Deckfarbenschmuck lieferte), vgl. Bl. 76r3 bzw. Bl. 5v mit Cod. 194, Bl. 27r (Ranken hier verkürzt zu zwei gegenständigen Blättern mit tropfenförmig verdickten Mitteladern); des Weiteren zeigt auch die Fleuronnée-Initiale Cod. 405, Bl. 124r, die von der in Sign. 14820 tätigen Hand stammt, starke Ähnlichkeiten zur Buchstaben körperfüllung der Ornamentalinitiale Cod. 405, Bl. 41v, sowie zum Binnenfeld Cod. 194, Bl. 27r. Ein wei terer Bezug zu Cod. 194 und zugleich Beleg für die Verbindung zw. Fleuronnée- und Deckfarbenhand ist die vierzeilige Fleuronnée-Initiale Sign. 14820, Bl. 47v3, die in der Art einer Ornamentalinitiale gestaltet wurde, wofür eine Deckfarbeninitiale wie Cod. 194, Bl. 45v (oder auch Cod. 405, Bl. 31v) als Vorlage gedient haben könnte (vgl. Rankenschlinge und Sitz des Goldpunktes). Der Versuch, eine Fleuronnée-Handschrift an die Farbigkeit eines mit Deckfarben ausgemalten Codex heranzuführen (durch Nuancierung: alle Farben auch in helleren bzw. blasseren Tönen eingesetzt), ist evtl. ein zusätzlicher Beleg dafür, dass der Florator auch Deckfarbendekor ausführte. S. R. E: Roter Samteinband über Holz. Neustift, 17. Jh. (?) Samt insbes. an den Ecken und Kanten etwas abgerieben, Holz vereinzelt freiliegend. Kanten gerade, nach innen abgeschrägt. Schnitt golden gefärbt. Am Spiegel des VD und HD sowie am Recto des Vor- und am Verso des Nachsatzbl. weiße Papiertapete mit Rankendekor aufgeklebt, am Übergang von Spiegel zu Vorbzw. Nachsatzbl. jeweils ein ca. 30 mm breiter Streifen schwarz gefärbt. Am Spiegel des VD Signaturschild (s. bei G). G: Das Neustifter Wappen Bl. 4v lässt darauf schließen, dass die Hs. in Neustift hergestellt wurde. Hs. bei vorübergehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. Am Spiegel des VD Neustifter Signaturschild 14.820 (= wohl Numerus currens der Neustifter Stiftsbibl.). L: Gschwend 86–88 (ohne Nennung der Sign.). – Lipphardt III 1086; VI 353, Nr. 629a (Sigle Neust5). – Gozzi 548, Nr. 82. – R. Kay, Pontificalia. A Repertory of Latin Manuscript Pontificals and Benedictionals. Lawrence 2007, 108, Nr. 559. – Baroffio 324, Nr. 16721. 340 Sign. 14820 (Ir–IIIr) leer. (IIIv) Inhaltsverzeichnis. 1 (IVr–VIIIr) Vier Evangelien bei der Fronleichnamsprozession (von späterer Hand, s. bei S). (VIIIv) leer. 2 (IXr–Xr) Gebete des Bischofs beim Anziehen der bischöflichen Gewänder (vgl. Bl. 76r–77v) (von späterer Hand, s. bei S). Gebet Ad caligas nur Titel, Text des Gebetes Ad calceos. (Xv) leer. 3 (1r–85v) Rituale Neocellense mit Breviertexten. Vgl. Cod. 194 und Cod. 726. (1r–2r) In nativitate dni (zusätzlich Ad laudes). (2r–3r) In circumcisione dni. (3r–4r) In epiphania dni. (4r–12r) In purificatione BMV. (5r) Benedictio candelarum. Tit.: In die sancto post tertias sequitur Benedictio Cereorum. – (10r) Processio candelarum. – (11r) Canticum Zachariae. (12r–13r) In annuntiatione BMV. (13r–24r) Dnca in palmis. (13r) In die palmarum ad processionem. – (15r) Exorcismus florum et frondium. – (15v) Benedictio palmarum. – (18v) Praefatio. – (21r) Aspersio et thurificatio palmarum. – Distributio palmarum. – (21v) In reditu. – (22v) Adoratio crucis. – (23v) Canticum puerorum. – (24r) In reditu ante altare oratio. (24v–27r) Fer. 5. in coena dni. (24v) In cena domini benedictio ignis. – (25r) Ad mandatum. (27v–35r) Fer. 6. in parasceve. (27v) Adoratio crucis. – (30r) Depositio crucis (Gschwend 86). – (30v) Vesper. (35r–43v) In vigilia paschatis (35r) Benedictio ignis. – (37r) In reditu canitur himnus Prudencii (AH 50, Nr. 31). – (37r) Ex sultet (nur Verweis). – (37r) Benedictio aquae. Mit Verweis auf Allerheiligenlitanei. (43v–46r) Dnca Resurrectionis dni. (Gschwend 86–88). (46r–47r) In vigilia ascensionis dni. (47r–48r) Benedictio aquae in vigilia Pentecostes. (48r–49r) In festo ss. Corporis Christi. (49r–50r) In assumptione BMV. (50v–51r) Augustinus. (51r–52r) In nativitate BMV. Eingeschoben: (51v) Adrianus (Ad Matutinas) und (51v) Corbinia nus (Ad Matutinas). (52r–53r) In dedicatione ecclesiae. (53r–v) De omnibus sanctis. Eingeschoben: (53v) Caesarius (Ad Matutinas). Sign. 14820 341 (54r–v) In commemoratione omnium fidelium defunctorum. Mehrere Gebete, davon eines Ad Completorium. Eingeschoben: (54r) In depositione beati Hartmanni. (55r–61r) Ordo paenitentiae. (61v–63r) Einkleidung der Novizen. Tit.: De indutione novitiorum. (63v–69r) Profess. (69r–70v) Ordo für die Aufnahme von Laien. Tit.: De fraternitate laicali. (70v–72v) Totenliturgie für die Konventualen. (72v) Datierung 1578. (73r–76r) Accessus altaris (vgl. Jungmann I 354–355). (76r–77v) Gebete des Bischofs beim Anziehen der bischöflichen Gewänder (vgl. Bl. IXr–Xr; Jungmann I 358ff.). (77v–78r) Bischöfliche Segensformel (Jungmann II 551). Tit.: Sequitur benedictio post missam. (78r–79r) Gebete nach der Messe (Jungmann II 571). (79v–83r) Collectae de tempore. Tit.: Sequuntur collecte seu orationes de tempore. Für Dnca 3., 4., 5. post epiph., Septuages., Sexages., Quinquages., Dnca 1., 2. in Quadrages., Dnca 3. mit Collectae feriales, Dnca 4. mit Collectae feriales, Dnca passionis mit Collectae feriales, Dnca 8.–16., 19.–22. post pent. (83r–85r) Collectae de sanctis. Tit.: Sequuntur Collecte de sanctis. Für Philippus et Iacobus, Monica, Iohannes ante Portam Latinam, Gordianus et Epimachus, Nicomedes, Marcellinus, Erasmus, Bonifatius, Primus et Felicianus, Barnabas, Basilides et Cyrinus et Nabor, Vitus et Modestus, Marcus et Marcellinus, Eusebius, Hermes, Undecim milia virginum, Severinus, Amandus, Simon et Iudas (mit Oratio in secundis vesperis), Narcissus. Vgl. Cod. 100, Bl. 97va–104ra mit ähnlicher Auswahl und Anordnung. (85r–v) Versikel. Tit.: Sequuntur nunc versiculi communes per annum. Enthält: Sabbatinus diebus ad vesperas, in secundis vesperis. De apostolis ad vesperas, ad matutinas. De martiribus ad vesperas, ad matutinas. De uno martire sive confessore ad vesperas, ad matutinas. De virginibus ad vesperas, ad matutinas. (85v) Schlussschrift: Laus Deo. Darunter Schreibernennung (?): F. H. 4 (85v–86r) Nachträge (von mehreren Händen, s. bei S). (85v) Drei Gebete: De s. Pangracio. Presta quesumus omnipotens Deus ut qui beati Pangratii martiris tui natalicia colimus … (vgl. Corpus orationum Nr. 4520a, für Valentin). Nereus et Achilleus. Semper nos domine martirum tuorum Nerei et Achillei foveat … (Corpus orationum Nr. 5465). De s. Wolffgango episcopo. Exaudi domine preces nostras quas in sancti Wolffgangi confessoris tui atque pontificis solemnitate deferimus … (vgl. Corpus orationum Nr. 2479, für Marcellus). (85v) Totengebet. Quesumus domine ut anime famuli tui cuius obitus sive diem commemoramus sanctorum tuorum largiri digneris consortium et rorem misericordie tue perennis infundere. Per. 342 Sign. 15063 (86r) Gebet. Deus cuius misericordie non est numerus suscipe propicius preces humilitatis nostre et animabus que nobis in oracionem commendate sunt et qua utimur vel usi suus elemosinis quibus tui nominis dedisti confessionem remissionem cunctorum tribue peccatorum. Per (Corpus orationum Nr. 1178). (86r) Oratio de s. Floriano. Deus qui nos annua beati Floriani martiris tui sociorumque eius gloriosa passionis solemnitate letificas … (vgl. Corpus orationum Nr. 1843, für Cyriacus). (86r) Totengebet in anniversario. Deus indulgentiarum domine da anime famuli (daneben: e) tui (daneben: e) cuius anniversarium depositionis diem et memoriam agimus refrigerii sedem quietis beatitudinem luminis claritatem. (86r) Versikel. Christus factus est pro nobis obediens usque ad mortem. (86r) Gebet. Domine Iesu Christe fili Dei vivi pone amaram mortem et passionem tuam inter animas nostras et misericordiam et districtum iudicium qui cum Deo patre et spiritu sancto vivis et (Corpus orationum Nr. 1251). (86v) leer. (Ir–IIv) leer. W. N. / U. S. Sign. 15063 ANTIPHONARIUM Perg. 197 Bl. 355 × 255. Wohl Neustift, 2. H. 15. Jh. (mit späteren Zusätzen). B: Perg. stark abgegriffen, vereinzelt kleine Löcher und Nahtspuren. Am unteren Blattrand bisweilen Fehl stellen. Bl. 58, 76, 77, 138 und 193 am Blattrand beschnitten. Lagen: (IV–1)7 + (IV–1)14 + (IV–1)21 + 5.IV61 + (IV–1)68+ IV76 + (IV–1)83 + (II–2)85 + III91 + (III–1)96 + (IV–1)103 + IV111 + (IV–1)117(118) + IV124(126) +III130(132) + 5.IV169(172) + (IV–2)175(178) + (I+3)180(181) + (IV–1)187(190) + (IV–3)192(195) + I194(197). Gegenbl. zu Bl. 7 als Spiegel auf dem VD aufgeklebt, Gegenbl. zu Bl. 8, 16, 65, 77, 84, 85, 95, 103, 114, 170, 171, 184, 188, 189, 190 herausgeschnitten (tw. Textverlust, s. bei Inhalt). Bl. 178–180 Einzelbl. Um die 14. und 19. Lage (Bl. 92 und 96, 125 und 130) Falz geschlagen, urspr. wohl Quaternionen. Am Lagenende bisweilen (Lagen 4–10, 17, 19, 21–23) Reklamanten. Am oberen Blattrand Foliierung 19. Jh.; Fehler in der Zählung: 116, 121 und 169 doppelt angeführt. Bl. 21 Spuren, Bl. 92 und 110 Reste von Ledersignakeln. S: Schriftraum 245/250 × 170/175, von zweifachen feinen Tinten- bzw. Blindlinien seitlich begrenzt: Spalte links jeweils für Notenschlüssel, Spalte rechts für Kustos verwendet. Zirkelstiche. Zehn rote Vierlinien systeme mit ebenso vielen Textzeilen auf feiner Tinten- bzw. Blindlinierung. Textualis formata (Textura). Zahlreiche Ergänzungen und Korrekturen von verschiedenen Händen in Tinte und Bleistift (bis ins 19. Jh.), zahlreiche Rasuren (s. bei Inhalt). N: Gotische Choralnotation (mit Virga) auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden. Reperkussionston mit einem ornamentalen Haarstrich. Vereinzelt schwarze Gliederungsstriche, insbes. am Schluss der einzel nen Gesänge. A: Rote Durchstreichungen und Überschriften. Rote und blaue Lombarden, selten mit Punktverdickungen, meist in der Höhe einer Notenzeile. Bl. 1r rote Lombarde mit einfacher Aussparung, zwei Notenzeilen hoch. Bl. 5r–v (Ergänzung) schwarze Initialbuchstaben. Schwarze Cadellen, eine Notenzeile hoch, mit rotem Füll strich (Bl. 1r und 2r Cadelle nachträglich weinrot konturiert) und schwarzem Fleuronnée von einer Hand, fallweise flüchtig ausgeführt: In den Binnenfeldern in der Regel Knospenformationen, bisweilen ausgesparte vereinfachte Blattformen oder Liniendekor; Konturlinien; als Besatz zumeist vereinfachter Perlenbesatz sowie häufig Profilköpfe, die an die Besatzköpfe des Hauptflorators der Neustifter Gradualien erinnern (Stirnwulst), Bl. 10v bärtiger Profilkopf, Bl. 53r weibliche Dreiviertelfigur. Auch bei den (späteren) Ergänzungen einfacher Dekor von mehreren Händen: 1) Bl. 81v–83v, 84v–95v und z. B. auch über Rasur Bl. 97r: abwechselnd rote und blaue (Blau pastos aufgetragen) Lombarden (I-Lombarde mit kopfstempelförmigem Besatzelement, z. B. Bl. 87v) mit Punktverdickungen; 2) Bl. 96v schwarzes Fleuronnée, ebenfalls den Hauptflorator der Neustifter Gradualien imitierend; diese Hand ergänzte Sign. 15063 343 möglicherweise auch Fadenausläufer (z. B. Bl. 12r Bas-de-page); 3) Bl. 97r2 blaue Lombarde (zwei Noten zeilen hoch) mit rotem Knospenfleuronnée, als Besatz Perlenpyramiden mit Fibrillen; 4) Bl. 193v rote Lom barde mit Punktverdickungen und Ausläufern, Cadellen mit rotem Füllstrich und etwas Dekor, jeweils eine Notenzeile hoch; 5) Bl. 194v schwarze Initialbuchstaben mit kritzeligem schwarzem Dekor (stilisierte Blätter). S. R. E: Renaissance-Einband: dunkelbraun gefärbtes Leder über Holz mit Blinddruck. Neustift, 2. H. 16. Jh., Werk statt: Narziß Schuechl. VD und HD gleich: sechs durch je zweifache Streicheisenlinien gebildete Rahmen (von außen nach innen): Rahmen 1 gefüllt mit Rolle Profilköpfe in Handspiegeln mit Quastenbehang (Nr. 1 = Neuhauser, Rollenein bände, R 22), Rahmen 2 gefüllt mit Rolle Eherne Schlange, Kreuzigung, Auferstehung (Nr. 2 = Neuhauser, Rolleneinbände, R 20), Rahmen 3 gefüllt mit Rolle Menschenpaare unter Baldachin mit floralem Dekor; am Ende der Rolle jeweils Umschrift DVRKO (Nr. 3 = Neuhauser, Rolleneinbände, R 45), Rahmen 4 gefüllt mit Einzelstempel Herzblatt (Nr. 4 = Neuhauser, Rolleneinbände, S 18 [?]) und Doppellilie mit rhombischem Mittelblatt (Nr. 5 = Neuhauser, Rolleneinbände, S 40), Rahmen 5 gefüllt mit Rolle florales Dekor (Nr. 6 = Neuhauser, Rolleneinbände, R 25), Rahmen 6 gefüllt mit Ranke mit eingeschlossener Fünfpunktblüte (Nr. 7 = Neuhauser, Rolleneinbände, R 23). Mittelfeld gefüllt mit Rolle Eherne Schlange, Kreuzigung, Auferste hung (Nr. 2). Jeweils fünf Beschläge, am VD und HD unterschiedlich: Am VD mittig rautenförmig mit ge radem Abschluss und mit mittig größerem und außen mehreren kleineren Buckeln. Eckbeschläge jeweils mit Spruchband ([…] mater […]), Mittelbeschlag und Blattwerkabschluss mit sechs kleineren Buckeln. Am HD mittig rautenförmiger Beschlag mit Blattornament, in den Ecken spitz zulaufende Beschläge mit floralem Dekor. Reste (oben) bzw. Spuren (unten) von zwei Leder-Metall-Schließen. Kanten gerade, nach innen hin abgeschrägt. Rücken: Häubchen oben und unten beschädigt; vier Doppelbünde, jeweils von zweifachen Streicheisenlinien gesäumt. Im untersten Feld Signaturschild 19. Jh. Kapitale, unten tw. beschädigt, mit na turfarbenem und rötlichem Zwirn umstochen. Spiegel des VD Pergamentblatt, beschrieben, unten tw. gelöst, Abklatsch auf Holz sichtbar, Spiegel des HD Pergamentblatt mit Federproben. G: Hs. bei vorübergehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck ge langt. Am Rücken Neustifter Signaturschild 15.063 (= wohl Numerus currens der Neustifter Stiftsbibl.). L: G. Gabrielli, Tropes in cantus planus sources from South Tyrol, in: J. Border u. a., Cantus Planus. Venice meeting 2014. Venezia (in Druck). – MeSch VI (in Vorbereitung), bei Wien, ÖNB, Cod. 2777 (Oswald von Wolkenstein, Liederhandschrift A) Sign. 15063 erwähnt (K. G. Pfändtner). Antiphonarium. 1 (VDS) Hymnus In adventu dni (AH 51, Nr. 47) mit Notation von späterer Hand. Darunter Ergänzung in Bleistift 19. Jh.: Versikel. Rorate Coeli desuper & nubes pluant iustum. Responsorium. Aperiatur terra et germinet salvatorem (Ies. 45,8). 2 (1r–58r) Temporale (Dnca 1. adventus bis In vigilia paschatis). (1r) Dnca 1. adventus. – (1v) Dnca 2. adv. – (2r) Dnca 3. adv. – (2v) O-Antiphonen (ab Bl. 3v, Z. 7 radiert). – (5r) In nativitate dni (z. T. von späterer Hand auf Rasur). Responsorium Descendit de celis mit Prosula (Hofmann-Brandt Nr. 416, CAO Nr. 6411); Responsorium Verbum caro mit Prosula (AH 34, Nr. 1, Hofmann-Brandt Nr. 537). – (13r) Stephanus. – Suff. (In nativitate BMV). – Zweistimmiger Tropus (B. Stäblein, Hymnen I: Die mittelalterlichen Melodien des Abendlandes [Monumenta Monodica Medii Aevi 1]. Kassel, Basel 1956, 396 [mit Transkription der Melodien]). – (14v) Expl. mut. (Antiphon BMV): … eius gloriam quasi uni[geniti (1 Bl. fehlt, s. bei B) (15r) Iohannes ap. Inc. mut. (Antiphon): ] michi ianuam vite et … – (15r) Innocentes. – (15v) Dnca infra octavam (tw. radiert). – (16v) In circumcisione dni. (Bl. 16v, 17v: tw. radiert und von späteren Händen neu geschrieben, Bl. 18r–19v mittig radiert). – (19v) In epiphania dni. – (20r) Dnca infra octavam epiph. – Per hebdomadam. – (21r) In octava epiph. – (21v) Dnca 1. post octavam epiph. – (22v) Fer. 2. – Fer. 3. – (23r) Fer. 4. – Fer. 5. – (23v) Fer. 6. – (24r) Dnca 2. post octavam epiph. – Dnca 3. post octavam epiph. – Dnca in Septuages. – (25v) Dnca in Sexages. – (26v) Dnca in Quinquages. – (27v) Dnca in Quadrages. – (29r) Fer. 2. (Notation nicht aus geführt). – Fer. 3. (Notation nicht ausgeführt). – Fer. 4. (Notation nicht ausgeführt). – (29v) 344 Sign. 15063 Fer. 5. – Fer. 6. – Dnca 2. in Quadrages. – (30v) Fer. 2. – (31r) Fer. 3. – Fer. 4. – Fer. 5. – Fer. 6. – Dnca 3. in Quadrages. – (31v–32r) am unteren Blattrand Ergänzung von drei Versen (Cantus Nr. 203225, CAO Nr. 3732zc und CAO Nr. 3732zb) mit Notation von späterer Hand zur An tiphon Media vita. – (32v) Fer. 2. – Fer. 3. – Fer. 4. – Fer. 5. – (33r) Fer. 6. – Dnca 4. in Quadrages. – (34r) Fer. 2. – Fer. 3. – Fer. 4. – (34v) Fer. 5. – Dnca in passione dni. – (35v) Fer. 2. – (36r) Fer. 3. – Fer. 4. – Fer. 5. – Fer. 6. – (36v) Dnca in palmis. – (37v) Fer. 2. – Fer. 3. – Fer. 4. – In coena dni (tw. radiert). Mit Nokturnen und Kyrie Qui passurus advenisti propter nos (CAO Nr. 8446,1). – (45v) Fer. 6. in parasceve. Mit Nokturnen. – (52r) Sabbato. – (58r) Ergänzung von späterer Hand: Antiphona de b. virgine (Salve sanctissima virgo Maria mater, nur Beginn mit metrischer Notation mit Zeichen der Mensuralnotation). (58v) nur Linierung. 3 (59r–84v) Sanctorale. (59r) Andrea. – (60r) Barbara: Reimoffizium (AH 25, Nr. 42). – (62r) Nicolaus. – (64r) Lucia. – (64v) Thomas Cant. (tw. radiert). Expl. mut. (Antiphon): … pravorum framea ce[(lum) (1 Bl. fehlt, s. bei B) (65r) [Fabianus et Sebastianus]. Inc. mut. (Antiphon): ] virum de plebe et claritatem … (tw. radiert). – (65v) Agnes (tw. radiert). – (66r) In conversione s. Pauli. – (67v) In purificatione BMV: Reimoffizium Videtur miraculum mater domini concepit (in AH nicht angeführt). Responsorium Videte miraculum mit zwei Tropen (Hoffmann-Brandt Nr. 281, CAO Nr. 7869 und Hoff mann-Brandt Nr. 262, CAO Nr. 7869zd). Mit Nokturnen (Bl. 71v, Z. 2–8 ohne Notation). – (72r) Agatha (fälschlich Agnetis v.). – (73v) Dorothea: Reimoffizium (AH 5, Nr. 56 [mit leichten Abweichungen]). – (76r) Ingenuinus et Albuinus (vgl. Breviarium Brixinense 1489, Bl. 271aβ). – (78r) Gregorius: Reimoffizium (AH 5, Nr. 64). – (79v) In annuntiatione BMV. (81v–84r, Z.1) Ergänzung von späterer Hand: (81v) In festo s. Michaelis archangeli. – (82v) In festo omnium sanctorum. (84r) Rudpertus. Inc. mut. (Antiphon): ] in populo Dei … – Expl. mut.: …viri sancti quod mole [ 4 (84v–95v) Ergänzung von späterer Hand: Commune sanctorum. (84v) De apostolis. – (85v) Temp. pasch. de apost. et martyr. – (87r) De uno martyre. – (88r) De martyribus. – (90r) De episcopo et confessore. – (91r) Pro doctore. – (91v) Pro solis sanctis pontificibus. – De communi confessore non pontifice. – (92v) De virginibus. – (94v) In dedica tione ecclesiae. (96r) Inc. mut. (Antiphon „Regina caeli“ mit Tropus Cantus Nr. 850222): si]cut dixit alleluia … – Marianische Antiphon. – (96v) Ergänzung von anderer Hand: De s. spiritu. Expl. mut. (Respon sorium): … et spiritui sancto [ (1 Bl. fehlt, s. bei B) 5 (97r–120v) Forts. des Temporale (Dnca resurrectionis dni bis In festo ss. corporis Christi). (97r) Dnca resurrectionis dni (tw. von späterer Hand auf Rasur). Mit Nokturnen. – (100r) Visita tio sepulchri (bei Lipphardt nicht angeführt; tw. von späterer Hand auf Rasur). – (102v) Fer. 2. – (103r) Fer. 3. – Fer. 4. – (103v) Fer. 5. (fälschlich Feria IIII). – Fer. 6. – Dnca 1. post pascha. – (104v) Dnca 2. post pascha. – (105r) Dnca 3. post pascha. – (106r) Dnca 4. post pascha (Dominica ultima). – (106v) In ascensione dni (tw. von späterer Hand auf Rasur). – (108v–109v, Z. 4) Rasur. – (109v) Dnca pentecostes et de spiritu sancto: nur Antiphon zum Magnificat der zweiten Vesper. – (110r) Fer. 2. – Fer. 3. – Fer. 4. – (110v) In festo ss. trinitatis (tw. von späterer Hand auf Rasur): Reimoffizium Gloria tibi trinitas equalis una Deitas (in AH nicht angeführt, Beginn mit Chev. Nr. 7287 übereinstimmend). – (112v) In festo ss. corporis Christi. Responso rium Homo quidam fecit cenam mit Prosula (Hofmann-Brandt Nr. 448, Cantus Nr. 601068Pa). Mit Nokturnen. 345 Sign. 15063 6 (120v–128r) Historiae aestivales. (120v) Liber Regum (tw. von späterer Hand auf Rasur). – (121v) Liber Sapientiae. – (123r) Historia Iob, unvollständig. (124v) Expl. mut. (Antiphon): … et peccata responde michi cur [ (1 Bl. fehlt, s. bei B) (125r) Liber Iudith, unvollständig. Inc. mut. (Responsorium): ]date cum omnes populi … – Liber Esther. – (125v) Liber Maccabaeorum. – (126v) Liber Ezechielis. 7 (128r–132r) Forts. des Temporale (Dncae post festum ss. trinitatis). (128r) Dnca 1. post trinitatem. – (128v) Dnca 2. – (129r) Dnca 3. – Dnca 4. – Dnca 5. – (129v) Dnca 6. – Dnca 7. – Dnca 8. – (130r) Dnca 9. – Dnca 10. – (130v) Dnca 11. – Dnca 12, unvoll ständig. Expl. mut. (Antiphon): … Dum transiret dominus [ (1 Bl. fehlt, s. bei B) (131r) Dnca 17. Inc. mut. (Antiphon): prin]cipis phariseorum sabbato manducare … – Dnca 18. – Dnca 19. – Dnca 20. – (131v) Dnca 21. – Dnca 22. – Dnca 23. – (132r) Dnca 24. – Dnca 25 (tw. radiert und gelöscht). 8 (132r–175v) Forts. des Sanctorale. (132r) Incipiunt hystorie de sanctis primo tempore paschali. Antiphonen und Responsorien aus dem Commune Sanctorum (tw. von späterer Hand auf Rasur). – (135r) Philippus et Iacobus. – (136r) In inventione s. crucis. – (137v) De lancea dni. Mit Hymnus (AH 52, Nr. 6) und Reim offizium (AH 5, Nr. 7). – (140r) Suffragia s. Alexandris. – Iohannes Baptista. – (141v) Iohannes et Paulus. – (142v) Petrus. – (144v) In octava Iohannis Baptistae. – (145r) In visitatione BMV. Mit zwei Tropen: 1) Responsorium Hec est ista (in Cantus nicht angeführt) mit Tropus Currens vini fluminis (in Hofmann-Brandt nicht angeführt); 2) Responsorium O preclara stella maris mit Prosula (AH 48, Nr. 399, Hofmann-Brandt Nr. 403). – (148v) Margaretha: Reimoffizium (AH 28, Nr. 3). Responsorium Quadam die Olibrius mit Prosula (Hofmann-Brandt Nr. 445). – (150v) Maria Magdalena. – (153r) Iacobus. – (153v) Petrus ad vincula. – (154v) Laurentius. – (155r) In assumptione BMV: Responsorium Que est ista que ascendit mit Prosula (Hofmann-Brandt Nr. 694). – (158v) Augustinus. – (162r) Suffragia Iohannis. – (162v) In nativitate BMV. – (164v) In exaltatione s. crucis. – (165r) In octava nativitatis BMV. – Matthaeus. – (166r) Suffragia Mauritii. – (166v) Suffragia Cosmi. – (167r) Michael (tw. radiert). – (168v) In translatione s. Augustini: Reimoffizium O rex altissime Deus mirabilis (bestehend aus zwei rhythmisierten Antiphonen, die eine ähnlich wie AH 25, Nr. 85 [Dominicus], die andere ähnlich wie Cantus Nr. 206563 [Monica]). – (169r) Gallus. – (169v) Ursula et undecim milia virginum. – (169bisv) Omnes sancti. – (170v) Martinus. – (172v) Suffragia Othmari. – Elisabeth: Reimoffizium (AH 25, Nr. 90). – (175r) Caecilia. – Clemens. – Catharina: Reimoffizium (AH 18, Nr. 37 und AH 26, Nr. 69). (175v) Expl. mut. (Antiphon): … et spiritui sanc[to (1 Bl. fehlt, s. bei B) 9 (176r–184r) Commune sanctorum. (176v) De apostolis. – (177v) De evangelistis. – (178v) De martyribus. – (180r) De uno martyre (tw. ohne Notation). – (183r) De virginibus (tw. ohne Notation). 10 (184r–185v) De conceptione BMV: Reimoffizium (AH 5, Nr. 13; Cantus Nr. 200447). 11 (185v–192v) Officium defunctorum. Mit Nokturnen (zweite Vesper ohne Notation, Responso rienreihe der Nokturnen nicht in K. Ottosen, The responsories and versicles of the Latin Office of the Dead. Aarhus 1993 angeführt). 12 (193r–194r) Marianische Gesänge (von verschiedenen späteren Händen). Mit Sequenz (AH 54, Nr. 201). 13 (194v) Ergänzung von späterer Hand: Antiphonae Sabbato per annum (nur Inc.). (HDS) Federproben, u. a. Specie tua et pulchritudine tua und Sancti tui domine florebunt. G. G. / U. S. 346 Nr. 139 (olim Sign. 14819) Nr. 139 (olim Sign. 14819) GRADUALE. ANTIPHONARIUM Pap. 231 Bl. 315 × 215. Umfeld Neustift, Ende 15. Jh. (T. I: 1495/96). B: Zwei Teile: I (1–153), II (154–231). T. I 1495/96 dat. (s. bei S), T. II nach Wasserzeichenbefund zeitnah entstanden. Wasserzeichen beider Teile z. T. übereinstimmend. Verwandte Wasserzeichen: u. a. in Neustift, Stiftsbibl., Nr. 940. Am unteren Blattrand Foliierung 20. Jh. (Bleistift, hier berücksichtigt). E: Gotischer Einband: helles Schweinsleder über Holz mit Blinddruck. Brixen oder Neustift, Ende 15. Jh. Leder abgerieben und insbes. am VD etwas fleckig. Am VD ein durch zwei- (oben) bzw. dreifache Streich eisenlinien gebildeter Rahmen, gefüllt mit Einzelstempel vierblättrige Blüte frei (Nr. 1) im Wechsel mit Adler in Rhombus (Nr. 2). Mittelfeld mit dreifachen Streicheisenlinien gerautet. In den vier Mittelfeldern Einzelstempel (stark abgerieben, nicht mehr erkennbar) in Kreis (Nr. 3), in den Außenfeldern vierblättrige Blüte (Nr. 4). An den Schnittpunkten Einzelstempel Lilie in konkavem Karo (Nr. 5). Aufschrift in Tinte 1496. Am HD ein durch dreifache Streicheisenlinien gebildeter Rahmen, gefüllt mit Einzelstempel sechs blättrige Blüte in Kreis (Nr. 6), seitlich rechts im Wechsel mit Lilie frei (Nr. 7). Mittelfeld mit dreifachen Streicheisenlinien gerautet. In den Feldern Einzelstempel fünfblättrige Blüte frei (Nr. 8). Auf den Schnitt punkten mittig Einzelstempel fünfblättrige Blüte frei (Nr. 8), ansonsten Dreiblatt frei (Nr. 9). Zwei LederMetall-Schließen, untere nur noch als Rest erhalten. Kanten gerade, unten und oben Einkerbungen im Holz. Rücken: blau gefärbt (wie öfters in Neustift), vier Doppelbünde. Unten aufgeklebtes Papiertitelschild, da runter ältere Signatur sichtbar (s. bei G). Kapitale mit Leder umschlagen, oben etwas beschädigt. Spiegel des VD und HD jeweils ein Pergamentblatt, am VD mit Besitzvermerk und Exlibris (s. bei G). G: Die Hs. stimmt in Inhalt und Aufbau weitestgehend mit Neustift, Stiftsbibl., Nr. 940 überein (vgl. auch ver wandte Wasserzeichen); möglicherweise stellt Nr. 139 eine Abschrift von Nr. 940 für die Verwendung in der Pfarre Kiens dar, geschrieben wohl vom selben Schreiber Heinrich Edlinger (vgl. Nr. 940). Am Spiegel des VD Besitzvermerk Ad ecclesiam […] V. M. […] parochiae Kiens, darüber Exlibris der Stiftsbibl. Neustift 18. Jh. Wann die Hs. aus der von Neustift betreuten Pfarre in die Stiftsbibl. gelangte, ist nicht bekannt. Hs. bei vorübergehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. Am Rücken Neustifter Signaturschild 14819 (= wohl Numerus currens der Neustifter Stiftsbibl.) und Signaturschild 20. Jh. 139. L: M. Peintner, Musikgeschichte des Stiftes, in: 850 Jahre Neustift. Katalog 94–100, hier 94 (ohne Nennung der Sign.). – Stefani 98. – Engels 309–310 (mit Abb. und Notenbeispielen). – G. Gabrielli, Tropes in cantus planus sources from South Tyrol, in: J. Border u. a. (Hrsg.), Cantus Planus. Venice meeting 2014. Venezia (in Druck). I (1–153) B: Pap. tw. stark verschmutzt, bes. an den Lagenanfängen und -enden, Bl. 56r am oberen Blattrand kreisförmi ger Fleck, elfte Lage stark vergilbtes Pap., Bl. 1 Wurmfraß, Bl. 1, 64, 109, 113 eingerissen. Bl. 3 und 6, 50–51 etwas kleiner (295/305 × 215). Lagen: (V–1)9 + 3.VI45 + (VI–2)55 + (V–1)64 + (VI–1)75 + (V–1)84 + 2.VI108 + 2.V128 + (VI+1)141 + VI153. Erste Lage mit Papierfalz umschlagen, in der Mitte der ersten Lage Per gamentfalz mit Resten einer Beschriftung. Gegenfalz zu Bl. 7 auf Bl. 3r aufgeklebt. Gegenbl. zu Bl. 48, 52, 68 herausgeschnitten (ohne Textverlust), an den Gegenfälzen Rahmung erhalten. Gegenfalz zu Bl. 56 auf Bl. 64v, zu Bl. 84 auf 76r aufgeklebt. Bl. 129 eingehängtes Einzelbl., Gegenfalz auf Bl. 141v aufgeklebt. Am Lagenende Reklamanten, meist stark beschnitten. Zw. Bl. 20 und 21 nachträglich eingehängtes kleines Einzelbl. (120 × 210), in Zählung (und Lagenformel) nicht berücksichtigt. Am unteren Blattrand alte Foli ierung (?) in arabischen Ziffern in Tinte nur mehr tw. sichtbar. Am rechten unteren Blattrand Oberlängen einer ehem. Beschriftung stark beschnitten. Am oberen Blattrand Foliierung wohl von der Hand des Schrei bers in roten römischen Zahlen von IIII–CIIC (= Bl. 3–153); Fehler in der Zählung: XVI, XIX, XXIV, XLIV, XLVI, XLIX, LII, LIII, LVII, LIX, LXII, LXVI, LXXII, LXXIV, LXXXII, LXXXIII, LXXXV, LXXXVI, LXXXVIII, XCI, XCIII, XCVIII, CVI, CVIII, CXI, CXIV, CXV, CXXII, CXXIII, CXXIV, CXXIX, CXXXII, CL, CLIII, CLVIII, CLXV, CLXVIII, CLXXXIV, CXCIII, CXCIV ausgelassen; C und CLXXI zwar ausgelassen, Blatt in der fortlaufenden Zählung jedoch berücksichtigt (heute Bl. 73 bzw. 129); LXXI doppelt angeführt. Bl. LXV falsch eingebunden, zw. Bl. LXXXIV und LXXXVII, heute als Bl. 63 gezählt. S: Schriftraum 185/200 × 125/140. Von zweifachen roten Tintenlinien gerahmt, zu sieben bis acht Vierliniensys temen sowie Textzeilen. In seitlicher Rahmung jeweils Notenschlüssel bzw. Kustos eingetragen. Textualis mit kursivem Einschlag, dat. 1495 (Bl. 1r) und 17. Juli 1496 (Bl. 54v). Schreiber: Heinricus Edlinger (vgl. Nr. 139 (olim Sign. 14819) 347 Nr. 940, Bl. 1r) (vgl. Krämer, Scriptores: wohl identisch mit Henricus Edling: München, BSB, Clm 6195, dat. 1482, Prov. Frauenzell). Bl. 123v Text in Bastarda (wohl von gleicher Hand). Vereinzelt Abschriften einzelner Wörter in Bleistift 19./20. Jh. Inhaltsverzeichnis (Bl. 1v–2v): Schriftraum 190/240 × 145/160. Vier Spalten, mit horizontaler Unterteilung für die Überschrift, jeweils von zweifachen roten Tintenlinien gerahmt, Textualis cursiva. Blätter zusätzlich von je zweifachen roten Tintenlinien am äußersten Blattrand gerahmt, urspr. wohl gesamte Hs. (vgl. Reste Bl. 3r unten, 4r oben), heute beschnitten. Bl. 1r Rahmung durch zweifache rote Tintenlinien mit Abgrenzung eines Binnenfeldes (175 × 95) und zu sätzlicher horizontaler Unterteilung. N: Gotische Choralnotation auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden, rote vertikale Trennstriche (lange Striche, d. h. über alle vier Notenlinien als Abschluss bei jedem Gesang; kurze Striche zw. den einzel nen Wörtern). Metrische Notation mit Zeichen der Mensuralnotation: Bl. 59v–60v Gloria Spiritus et alme unter Verwen dung von Semibreves, Breves (gebildet aus zwei aneinandergefügten Semibreves) und Minimae (mit rotem Notenhals). Beim Tropus Rubriken chorus und pueri; Bl. 118v–121v, 123v: vier Credos und eine Sacra Cantio unter zusätzlicher Verwendung von Semiminimae. A: Rote Paragraphzeichen, Auszeichnungsstriche, Verbindungsstriche (vereinzelt auch in Mintgrün), Trenn striche (bei Notation), Unterstreichungen, Überschriften. Oberlängen und Unterlängen vereinzelt cadellen artig verziert. Rote und tintenfarbene, vereinzelt auch mintgrüne Initialen mit Aussparungen (vereinzelt kopfstempelförmig), Punktverdickungen und einfachen Ausläufern, bisweilen mit einzelnen Punkten, ein ander gegenüberstehenden Punkten oder auch Dreipunktmotiv am Buchstabenschaft und vereinzelt auch im Binnenfeld, bisweilen mit Ausläufern in Achterform gelegt, tw. gestrichelt (Bl. 118v, 135r), tw. mit Punkten versehen (Bl. 129r). Bl. 69v Ausläufer als Band gestaltet. Tintenfarbene Initialen mit roter Tinte umrahmt oder verziert, meist cadellenartig gestaltet. Bl. 109r rote Initialen mit mintgrüner Farbe nachgefahren. Bl. 126v am unteren Blattrand rote Tinte verschmiert/verwischt. Graduale. (1r) Datierung in Rot: Anno Cristi filii Dei millesimo quadringentesimo novagesimo quinto die comme moracionis s. Pauli. Darunter später in Schwarz: anno 1496, darunter korr. 1495. Todesnotiz in Rot: Heinricus Edlinger devixit anno Christi 1496 in die s. Steffani (26. Dezember 1496). Darunter in Schwarz abgeschrieben 19. Jh. Henricus Edlmor (!) devixit anno Christi 1496 in die s. Stephani. (1v) nur Rahmung. (2ra–va) Inhaltsübersicht mit Blattangabe. 1 (3r–54v) Temporale. (3r) Dnca 1. adventus. – (4r) Dnca 2. adv. – (5r) Dnca 3. adv. – (6v) Dnca 4. adv. – (7v) In galli cantu. Mit Sequenz (AH 53, Nr. 16). – (8v) In aurora. Mit Sequenz (AH 53, Nr. 15 mit propter beatam virginem divisio). – (11v) In nativitate dni (publica missa). Mit Sequenz (AH 53, Nr. 16). – (14v) Stephanus. Mit Sequenz (AH 53, Nr. 215). – (18r) Iohannes ev. Mit Sequenz (AH 53, Nr. 168). – (20v) Innocentes. Mit Sequenz (AH 53, Nr. 157). Mit eingehängtem kleinen Einzelbl. 20I. – (22v) In circumcisione dni. Mit Sequenz (AH 53, Nr. 16). – (24v) In epiphania dni. Mit Sequenz (AH 53, Nr. 29). – (27v) In coena dni. – (29r) Dnca resurrectionis dni. Mit drei Sequenzen (AH 53, Nr. 50; AH 54, Nr. 148; AH 54, Nr. 7). – (34r) In rogacionibus. – (35v) In ascensione dni. Mit Sequenz (AH 53, Nr. 67). – (38r) Dnca pentecostes et de spiritu sancto. Mit Sequenz (AH 53, Nr. 70). – (42r) In festo ss. trinitatis. Mit Sequenz (AH 53, Nr. 81). – (44v) In festo ss. corporis Christi. Mit Sequenz (AH 50, Nr. 385 mit divisio), Offertorium Sacerdotes mit Prosula Ad panem vite nunc accedite und zusätzlich zum Schluss Alia prosa (anderer Text für dieselbe Melodie: AH 49, Nr. 643). – (49v) In dedicatione ecclesiae. Mit Sequenz (AH 53, Nr. 247). – (52r) Pro defunctis. Mit drei Communiones (CAO Nr. 1211, CAO Nr. 1420 und Can tus Nr. 204995). Darunter (Bl. 54v): Anno 1496 in beati Allexii profesto (= 17. Juli). (55r–v) nur Rahmung und Linierung. 348 Nr. 139 (olim Sign. 14819) 2 (56r–80v) Missae BMV. (56r) Ohne Tit. Mit Sequenz (AH 54, Nr. 218), Offertorium Recordare mit Prosula (AH 49, Nr. 624). – (59r) In summo festo b. Marie. Kyrie, Gloria rhythmisiert mit Tropus Spiritus et alme orphanorum paraclite (Bosse Nr. 49). Anschließend Aliud et mediocre: Kyrie, Gloria mit Tropus Spiritus et alme (Bosse Nr. 23). – (63r) Aliud de virginibus. Kyrie. – Aliud ad idem. Kyrie. – (63v) Alleluia Ave benedicta Maria mit Tropus (Cantus Nr. 507047), Sequenz (AH 54, Nr. 218). – (64v) Tit.: Sequitur de s. Maria in adventu domini. Mit Sequenz (AH 54, Nr. 191). – (67v) Infra nativitatem et purificationem. Mit zwei Sequenzen (nur Verweis auf Gaude Dei genitrix, Divisio von AH 53, Nr. 15 und AH 54, Nr. 2). – (69r) In purificatione BMV: Ante processionem a ntiphona (Cantus Nr. 200456). – (69v) Ad missam. Mit Sequenz (AH 53, Nr. 99). – (73r) In annuntiatione BMV. – (73r) Circa festa pascalia. Mit Sequenz (AH 54, Nr. 18). – (74v) In assumptione BMV. Mit Sequenz (AH 53, Nr. 104). – (76v) In nativitate BMV. Mit Sequenz (AH 53, Nr. 95). – (78r) In visitatione BMV. Mit zwei Sequenzen (AH 54, Nr. 193; AH 54, Nr. 219 mit Rubrik: Generalis). 3 (80v–108v) Sanctorale. Tit.: Sequitur modo de sanctis per circulum anni. (80v) Iohannes Baptista. Mit Sequenz (AH 53, Nr. 163). – (83r) Petrus et Paulus. Mit Sequenz (AH 53, Nr. 210). – (85r) Paulus. – (85v) Maria Magdalena. Mit Sequenz (AH 50, Nr. 268). – (88v) Laurentius. Mit Sequenz (AH 53, Nr. 173). – (90v) Michael. Mit Sequenz (AH 53, Nr. 192). – (93v) Omnes sancti. Mit Sequenz (AH 53, Nr. 112). – (94v) Martinus. Mit Sequenz (AH 53, Nr. 181 mit Divisio propter simplices conf.). – (97r) Elisabeth. Mit Sequenz (AH 55, Nr. 120). – (98v) Catharina. Mit Sequenz (AH 54, Nr. 52). – (100v) Andreas. Mit Sequenz (AH 53, Nr. 122). – (102r) Nicolaus. Mit Sequenz (AH 55, Nr. 265). – (105r) In cathedra s. Petri. – (105v) De quodlibet sancto post septuagesimam. – (106r) De uno beato post pasca vide licet Georgio vel alio. Mit Verweis auf Sequenz (AH 54, Nr. 7). – (107r) Philippus et Iacobus. Mit Verweis auf Sequenz (AH 54, Nr. 7). – (108r) In inventione s. crucis. Mit Verweis auf Sequenz (AH 54, Nr. 7). 4 4.1 (109r–123v) Kyriale. (109r–118v) Kyrie. Gloria. (109r) In summis festis. Kyrie mit Tropus (AH 47, Nr. 99, Str. 1a, 2a und 3a). – (109r) Aliud in festo summo. Kyrie mit Tropus (AH 47, Nr. 5, Str. 1a, 2a und 3c). – (109v) Angelicum sive solemne. Kyrie, Gloria. – (111v) Pascale. – (112v) De apostolis. Nur Kyrie. – (113r) De confes soribus. – (114v) De martiribus. – (116r) De virginibus. – (117r) Dominicale. 4.2 (118v–121v) Credo. (118v) Tit.: Symbolum concilii Nyceni. – Patrem omnipotentem … et sepultus est (Miazga Nr. 450, in C statt in F; in C auch in Innsbruck, ULB Tirol, Cod. 457, Bl. 90r). – (119v) Patrem omnipotentem … et homo factus est (bei Miazga nicht angeführt). – (120r) Aliud. – Patrem omni potentem … et homo factus est (Miazga Nr. 84). – (120v) Aliud. – Patrem omnipotentem … et sepultus est (Miazga Nr. 123). 4.3 (121v–123v) Sanctus. Agnus Dei. (121v) Angelicum. Nur Sanctus. – (122r) Solenne. Sanctus mit Tropus (AH 47, Nr. 382, Str. 1a). – (122v) Ohne Tit. Nur Sanctus. – (123r) Paschale. 5 (123v) Sacra cantio. Rubrik: Infra elevacionem canendum est hoc. – Iesus Christus nostra salus (rhythmisiert) (AH 45, Nr. 125). 6 (124r–125v) Officium missae contra pestilentiam. Nr. 139 (olim Sign. 14819) 349 7 (125v–129r) Forts. des Kyriale: Sanctus. Agnus Dei. (125v) De beata virgine Maria. Nur Sanctus. – (126r) Aliud de eadem. Nur Sanctus. – (126v) De virginibus. – (127r) De martiribus. – (127v) De confessoribus. – (128v) In adventu dni. – (129r) In festis generale. Nur Sanctus (unvollständig). (129v) nur Rahmung und Linierung. 8 (130r–153r) Commune sanctorum. Jeweils mehrere Introitus, Gradualia, Alleluia, Offertoria und Communiones angeführt. (130r) De apostolis. Mit Sequenz (AH 53, Nr. 228). – (133r) De martyribus. Mit zwei Sequenzen (AH 55, Nr. 14; AH 53, Nr. 229). – (138r) De uno martyre. Mit Sequenz (AH 55, Nr. 9) und zwei Communiones (die zweite Communio mit zwei unterschiedlichen Melodien). – (142v) De confessore et pontifice. – (143r) De simplici confessore. Mit zwei Sequenzen (AH 54, Nr. 88; AH 53, Nr. 243). – (147v) De virginibus. Introitus mit Angabe von zwei zusätzlichen Versen: 1) De omnibus sanctis; 2) In assumptione BMV. – (148r) De virginibus (!). Mit Sequenz (AH 50, Nr. 271) und zwei Offertoria mit unterschiedlicher Melodie (maius, minus). (153v) nur Rahmung und Linierung. II (154–231) B: Bl. 154r stark verschmutzt, Bl. 170v–171r Tintenklecks, Bl. 171v Tintenspritzer. Bl. 206 und 207 etwas kleiner (295 × 215). Lagen: (VI–1)164 + 5.VI224 + (IV–1)231. In der Mitte der letzten Lage Pergamentfalz. Am unteren Blattrand alte Foliierung (?) in arabischen Ziffern in Tinte tw. beschnitten, tw. sichtbar. Am unteren Blattrand Reste einer ehem. Beschriftung stark beschnitten, tw. sichtbar. Am oberen Blattrand zeitgenössi sche Foliierung in roten römischen Zahlen von I–XCII (= Bl. 154–224); Fehler in der Zählung: X, XXVI, XXVII, XXIX–XXXIX, XLV, XLVI, LIII, LVII, LXIV, XCI ausgelassen; LXXXVII zwar ausgelassen, Blatt in der fortlaufenden Zählung jedoch berücksichtigt (heute Bl. 221). S: Schriftraum 190/205 × 130/140. Von zweifachen roten Tintenlinien gerahmt, zu jeweils 7–8, Bl. 223v 9 Vier liniensystemen samt Textzeilen. Textualis mit kursivem Einschlag, von gleicher Hand wie T 1. Bl. 220v und 221r–v. Am äußersten seitlichen Blattrand Notizen in Bastarda, beschnitten. Bl. 220v–223v am seitlichen Blattrand Bleistiftnotiz 19./20. Jh. Inhaltsverzeichnis (Bl.154r–v) Schriftraum 185/200 × 150. Vier Spalten, mit horizontaler Unterteilung für die Überschrift, jeweils von zweifachen roten Tintenlinien gerahmt, Textualis cursiva (wohl gleiche Hand wie Schreiber). Blätter zusätzlich von je zweifachen roten Tintenlinien am äußersten Blattrand gerahmt. Auch Bl. 155r zusätzlicher äußerer Rahmen erhalten. N: Gotische Choralnotation auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden, rote vertikale Trennstriche (lange Striche, d. h. über alle vier Notenlinien als Abschluss bei jedem Gesang; kurze Striche zw. den einzel nen Wörtern). A: Rote Paragraphzeichen, Auszeichnungsstriche, Verbindungsstriche (vereinzelt auch in Mintgrün), Trenn striche (bei Notation), Unterstreichungen, Überschriften. Rote, tintenfarbene und mintgrüne Initialen mit Aussparungen (vereinzelt kopfstempelförmig), Punktverdickungen und einfachen Ausläufern, bisweilen mit einzelnen Punkten, einander gegenüberstehenden Punkten oder auch Dreipunktmotiv am Buchstabenschaft und vereinzelt auch im Binnenfeld, bisweilen mit Ausläufern in Achterform gelegt, an den Schnittstellen tw. gestrichelt (Bl. 159v, 166v, 174r in der Gegenfarbe), tw. mit Punkten versehen. Bl. 164v Initiale D Buch stabenkörper mit Punkten versehen, am Schaft zwei einander gegenüberstehende Punkte, Fadenausläufer nach unten und oben, nach links Ausläufer in Form eines zehnlinigen Notensystems, mit einzelnen Noten (Punkten) versehen. Antiphonarium. (154r–v) Register. (154r) Tit.: Registrum Antiphonarii. Am unteren Blattrand Federprobe Notation. (154v) Inhaltsübersicht mit Blattangabe. 350 Nr. 139 (olim Sign. 14819) 1 (155r–191v) Temporale und Sanctorale, gemischt. (155r) Dnca 1. adventus. – (155v) Dnca 2. adv. – (156r) Dnca 3. adv. – (157r) Dnca 4. adv. – (157v) In die Lucie an. O (O-Antiphon). – (158r) Dnca 2. (O-Antiphon). – Dnca 3. (O-Antiphon). – Ohne Tit. (O-Antiphon). – (158v) In sancta nocte nativitatis dni. Responsorium Descendit de caelo, Vers Tamquam sponsus mit Prosula (Hofmann-Brandt Nr. 416, CAO Nr. 6411). – (167r) De BMV. – (168v) Stephanus. – (168v) Maria. – (169r) Iohannes. – (169r) Innocentes. – (169v) Dominica infra [octavam]. – In circumcisione dni. – (170r) In epiphania dni. Responsorium Illu minare illuminare Ierusalem mit Prosula (Hofmann-Brandt Nr. 673). – (172r) Paulus. – (172v) In purificatione BMV. – (173v) In annuntiatione angelica. – (174v) In nocte sancti pascatis. – (179r) Ad visitationem sepulchri (Engels 310 [mit Transkription]). – (179v) Ad laudes antiphona. – Angelus autem domini descendit de celo et accedens … – (180r) Ad Benedictus antiphona. – Et valde mane una sabbatorum … – (180v) In secundis vesperis. – Et dicebant ad inficem (!) quis revolvet nobis lapidem … – (181v) Alia antiphona. – Isti sunt sermones quos dicebam vobis … – (182r) In ewangelio. – Cum esset sero die illa una sabbatorum … – (183r) In vigilia ascensionis dni. – (183v) In ascensione dni. – (184v) De sanctis infra pascha et pentecostes. – (185v) In inventione s. crucis. – (187r) Iohannes ante Portam Latinam. – In nocte sancta pentecostes. – (188r) In festo ss. trinitatis: Reimoffizium (LMLO XT31). – (189r) In festo ss. corporis Christi. Responsorium Homo quidam fecit cenam mit Prosula (Hofmann-Brandt Nr. 448). 2 (191v–194r) Historiae aestivales. (191v) Tit.: Incipiunt hystorie estivales. (191v) Et primo de libris Regum. – (192r) Hystorie Sapiencie. – (192v) Hystoria Iob. – (193r) Hystoria Thobie. – (193r) Iudith et Hester. – (193v) Hystoria Machabeorum. – (194r) Hystoria Ezechielis. 3 (194v–210v) Forts. des Sanctorale. (194v) Iohannes Baptista. – (195r) Petrus et Paulus. – (196r) Margaretha: Reimoffizium (AH 28, Nr. 3). Responsorium Quadam die Olibrius mit Prosula (Hofmann-Brandt Nr. 445). – (197v) Maria Magdalena. Responsorium Accessit ad pedes mit Prosula (Hofmann-Brandt Nr. 696). – (199r) Laurentius. – (199v) In assumptione BMV. – (201v) In nativitate BMV. – (203v) Michael. – (204v) Omnes sancti. – (205v) Martinus. – (207r) Catharina. Antiphon zum Magnificat (AH 18, Nr. 37 und AH 26, Nr. 69). Responsorium Surge virgo mit Prosula (Hofmann-Brandt Nr. 483). – (208r) Andreas. – (209r) Nicolaus. – (210r) Thomas. (211r–212v) nur Linierung. 4 (213r–220v) Commune sanctorum. (213r) Commune apostolorum. – (213v) De evangelistis. – (214v) De pluribus martyribus. – (215v) De uno martyre. – (216v) De confessore et pontifice. – (217r) De simplici confessore. – (218r) De una virgine vel electa. – (219r) In dedicatione ecclesiae. 5 (220v–221v) Differentiae et Toni psalmorum. (220v) Tit. (am Blattrand von späterer Hand ergänzt): Versus memoriales pro octo tonis. Über sicht (Summula tractatus metricus de musica glossis commentarioque instructus, ed. E. Vetter [Divitiae musicae artis, A/VIIIa]. Buren 1988, 89). – (220v) Primus tonus. – (221r) Secundus tonus. Tercius. Quartus. Quintus. – (221v) Sextus. Septimus. Octavus. 6 (221v–223r) Melodien für Benedicamus Domino. (221v) Solenniter. Pascaliter. – (222r) Medium. Simplex cum tribus. De corpore Christi. Mit Tropus Benedicamus in laude panis … – (222v) Minus. De beata virgine Maria. Mit Tropus (Cantus Nr. a00821). – De apostolis. De martiribus. De confessoribus. De virginibus. Mit Tropus Benedicamus ad honorem virginis pie Katherine … – (223r) De apostolis. Mit Tropus Apostolo rum solennia sacrata … Laudetur sancta trinitas … Nr. 940 351 7 (223r–224r) Melodien für Ite missa est. (223r) Solenniter. Mit Tropus (AH 47, Nr. 481). – (223v) De BMV. Mit Tropus (AH 47, Nr. 487). – De s. Maria. Mit Tropus (MMMA XIX Nr. 221). – De apostolis. Mit Tropus (AH 47, Nr. 475). 8 (223v–224v) Zwei Antiphonen: De BMV, De omnibus sanctis. (225r–228r) nur Rahmung und Linierung. (228v–231r) nur Rahmung. (231v) nur Rahmung und Linierung. G. G. / U. S. Nr. 940 GRADUALE. ANTIPHONARIUM Pap. I, 229, I* Bl. 310 × 220. Umfeld Neustift, 1490/93. B: Zwei Teile: I (I, 1–151), II (152–229, I). T. I dat. 1490/93 (s. bei S), T. II nach Wasserzeichenbefund zeitnah entstanden. Wasserzeichen beider Teile z. T. übereinstimmend. Verwandte Wasserzeichen: u. a. in Neustift, Stiftsbibl., Nr. 139 (olim Sign. 14819). Am unteren Blattrand Bleistiftzählung 21. Jh. (hier berücksichtigt). E: Gotischer Einband: braunes Leder über Holz mit Blinddruck. Brixen oder Neustift, Ende 15. Jh. Leder insbes. am HD tw. abgerieben und beschädigt, am Rücken oben eingerissen. Am VD ein durch zwei- (unten) bzw. dreifache Streicheisenlinien begrenzter Rahmen, gefüllt mit drei einander abwechselnden Einzelstempeln vierblättrige Blüte frei (Nr. 1), gebogener Ast (Nr. 2) und punktiertes Herz frei (Nr. 3). Mit telfeld von zwei- bzw. dreifachen Streicheisenlinien gerautet. In den vier Mittelfeldern am VD Einzelstempel sechsblättrige Blüte in Kreis (Nr. 4), in den Außenfeldern rautenförmige Lilie frei (Nr. 5). Auf den Schnitt punkten Einzelstempel Punktdekor in Karo (Nr. 6). Am HD ein durch dreifache Streicheisenlinien begrenz ter Rahmen, gefüllt mit Einzelstempel Astdekor in Rechteck (Nr. 7). Mittelfeld mit dreifachen Streicheisen linien gerautet. In den vier Mittelfeldern Einzelstempel Osterlamm mit Fahne in Kreis (Nr. 8), in den Außenfeldern Nr. 5. Auf den Schnittpunkten Einzelstempel Vierblatt in konkavem Karo (Nr. 9). Jeweils drei der urspr. vier spitz zulaufenden Eckbeschläge mit kleinem halbrundem Buckel erhalten, von einstigem rautenförmigen Mittelbeschlag nur Nägel tw. erhalten. Zwei Leder-Metall-Schließen, obere nur mehr tw. erhalten, unten Leder beschädigt, Lager und Raster mit Aufschrift Maria. Kanten gerade, unten, seitlich und oben jeweils leichte Einkerbungen im Holz. Rücken: vier Doppelbünde, durch Streicheisenlinien betont. Felder zw. den Bünden jeweils mit zweifachen Streicheisenlinien mittig unterteilt. Im vierten Feld auf geklebtes Signaturschild (s. bei G). Kapitale mit naturfarbenem, grünem und rötlichem Garn umstochen, oberes Kapital leicht beschädigt. Spiegel des VD und HD jeweils ein Papierblatt, am VD leer, am HD mit Notiz (s. bei Inhalt). G: Hs. von dem aus Passau stammenden Heinrich Edlinger für die Verwendung in der Diözese geschrieben (vgl. Bl. 1r). Die Hervorhebung des hl. Ulrich sowie die später ergänzten Gebete am Spiegel des HD lassen auf eine Verwendung in der Kirche St. Ulrich in Ilstern (Pfarrei St. Sigmund / Kiens) schließen; Entstehung möglicherweise in Zusammenhang mit Neuerrichtung der Kirche 1491. Wann die Hs. aus der von Neustift betreuten Pfarre in die Stiftsbibl. gelangte, ist nicht bekannt. Hs. bei vorübergehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. Am Rücken Neustifter Signaturschild 20. Jh. 940. Die Hs. stimmt in Inhalt und Aufbau weitestgehend mit Neustift, Stiftsbibl., Nr. 139 (olim Sign. 14819) überein (vgl. auch verwandte Wasserzeichen); möglicherweise stellt Nr. 139 eine Abschrift von Nr. 940 für die Verwendung in der Pfarre Kiens dar. I (I, 1–151) B: Pap. tw. verschmutzt, Bl. 11, 20 sowie Bl. 103–123 an der rechten unteren Ecke große Tintenflecke, Pap. dadurch auch tw. beschädigt und eingerissen, Bl. 105 Pap. seitlich eingerissen. Lagen: 1I (Vorsatzbl., Perg.) + 2.V20 + VI32 + 5.V82 + VI94 + IV102 + V112 + (VI–1)123 + 2.V143 + IV151. Vorsatzbl. eingehängtes Einzelbl. In 352 Nr. 940 der Mitte der ersten Lage Pergamentfalz. Gegenbl. zu Bl. 123 herausgeschnitten (ohne Textverlust). Lagen reklamanten mittig, tw. stark beschnitten, vereinzelt auch Seitenreklamanten am Blattrand (vgl. z. B. Bl. 54r, 57r, 61r, 68r). Am Lagenanfang Kustoden (Ordinalzahlen), stark beschnitten. Am oberen Blattrand Foliie rung wohl von der Hand des Schreibers in roten römischen Zahlen von I–CLXXV; Fehler in der Zählung: XI, XII, XIII, XV, XVIII, XXVIII, XXXVII, XLV, L, LII, LVII, LXII, LXVIII, LXXIX, LXXXII, LXXXVI, XCV, XCVI, CVIII, CIX, CXVIII, CXIX, CXXXI, CXXXIII, CXLV und CLXVII ausgelassen; XVI und LXXX doppelt angeführt; VIII und XXII zwar nicht angeführt, Blätter in der fortlaufenden Zählung jedoch berücksichtigt. S: Schriftraum 180/205 × 120/140. Von zweifachen roten Tintenlinien gerahmt, zu sieben Vierliniensystemen sowie Textzeilen. In seitlicher Rahmung jeweils Notenschlüssel bzw. Kustos eingetragen. Textualis mit kur sivem Einschlag. Schreiber: Heinricus Edlinger, gest. 26. Jänner 1496 (vgl. Nr. 139 [olim Sign. 14819], Bl. 1r) (Bl. 1r), dat. 1490 (Bl. 1r) bzw. 1493 (Bl. 2r) (vgl. Krämer, Scriptores: wohl identisch mit Henricus Edling: München, BSB, Clm 6195, dat. 1482, Prov. Frauenzell). Inhaltsverzeichnis (Bl. 1v–2v): Schriftraum ca. 250 × 150. Vier Spalten, jeweils von zweifachen roten Tinten linien gerahmt, mit zusätzlicher horizontaler Unterteilung für die Überschrift. Textualis cursiva (ebenso von Henricus Edlinger). Blätter zusätzlich von je zweifachen roten Tintenlinien am äußersten Blattrand gerahmt, tw. beschnitten, urspr. wohl gesamte Hs. (vgl. auch Bl. 3r). Bl. 1r Rahmung durch zweifache rote Tintenlinien mit Abgrenzung eines Binnenfeldes (170 × 95) und zu sätzlicher horizontaler Unterteilung. N: Gotische Choralnotation auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden, rote vertikale Trennstriche (lange Striche, d. h. über alle vier Notenlinien als Abschluss bei jedem Gesang; kurze Striche zw. den einzel nen Wörtern). Metrische Notation mit Zeichen der Mensuralnotation: Bl. 23r–v Prosula Triumphat Dei filius unter Verwen dung von Semibreves, Breves (gebildet aus zwei aneinandergefügten Semibreves) und Minimae (mit rotem Notenhals). A: Rote Paragraphzeichen, Auszeichnungsstriche, Verbindungsstriche (Bl. 112v auch in Mintgrün), Trenn striche (bei Notation), Unterstreichungen, Überschriften. Oberlängen und Unterlängen vereinzelt cadellen artig verziert. Rote und tintenfarbene (Bl. 112v auch mintgrüne) Initialen mit Aussparungen (vereinzelt kopfstempelförmig), Punktverdickungen und einfachen Ausläufern, bisweilen mit einzelnen Punkten, einander gegenüberstehenden Punkten oder auch Dreipunktmotiv am Buchstabenschaft und vereinzelt auch im Binnenfeld, bisweilen mit Ausläufern in Achterform gelegt, vereinzelt gestrichelt (Bl. 83r, 113v), ver einzelt mit Punkten versehen (Bl. 112v). Tintenfarbene Initialen mit roter Tinte umrahmt oder verziert, meist cadellenartig gestaltet. Graduale. (Ir) Tit. (rot): Graduale sancti Udalrici. (Iv) leer. (1r) Datierung und Schreibernennung: (rot:) Anno Cristi celitus missi (schwarz:) millesimo quadrin gentesimo nonagesimo conscriptus est presens libellus cantualis ad divinum cultum augmentandum (gestrichen: conscriptus) per Heinricum Edlinger presbiterum Pataviensis diocesis. Cuius inicium dies prima Aprilis habuit feliciter secundum chorum Brixinensis ecclesie. Ut videri habet registrum et rubrica subsequentes in speciali de maioribus festis et ceteris minoribus agentes. Primo Graduale deinde Antiphonarium exequentes. (1va–2ra) Inhaltsübersicht mit Blattangabe. Tit.: Registrum Gradualis in presenti libello contenti iuxta numerum foliis assignatum. Darunter Datierung (rot): 1493. (2va–b) nur Rahmung. 1 (3r–55r) Temporale. (3r) In galli cantu. Mit Sequenz (AH 53, Nr. 16). – (4r) In nativitate dni (Ad missam publicam). Mit Sequenz (AH 53, Nr. 16). – (7r) Stephanus. Mit Sequenz (AH 53, Nr. 215). – (10r) Iohannes ev. Mit Sequenz (AH 53, Nr. 168). – (12v) Innocentes. Mit Sequenz (AH 53, Nr. 157). – (15r) In circumcisione dni. Tit.: In circumcisione domini aut de sancta Maria infra nativitatem Christi et purificacionem eiusdem Marie. Ad missam. Mit Verweis auf Sequenz Gaude Dei genitrix Nr. 940 353 (Divisio von AH 53, Nr. 15) und Sequenz (AH 53, Nr. 15, mit Divisio). – (18r) In epiphania dni. Mit Sequenz (AH 53, Nr. 29). – (21v) Rubrik: De accionibus fiendis modo vide alibi. – (21v) In die pasce ante processionem. – (22r) Ad processionem. Antiphon Cum rex glorie (Cantus Nr. 201042) mit Prosula (Cantus textualis) Triumphat Dei filius in metrischer Notation (Lipp hardt II 526–527, Nr. 385a, Z. 37–39 und 43–46). – (23v) Cantus puerorum (Salve festa dies …, AH 50, S. 79 [o. Nr.]). Mit Angabe der Variationen für Christi Himmelfahrt und Pfingsten. – (24r) Ad chorum (Prozessionsantiphon). – (24v) Dnca resurrectionis dni. Mit drei Sequenzen (AH 54, Nr. 7; AH 53, Nr. 50; AH 54, Nr. 148). (30v) De uno martire post pasca. Mit Verweis auf Sequenz (AH 54, Nr. 7). – (31v) Philippus et Iacobus. – (32v) De s. cruce. Mit Verweis auf Sequenz (AH 54, Nr. 7). – (34r) In rogacionibus ad processionem. – (35v) In ascensione dni. Mit Sequenz (AH 53, Nr. 67). – (38v) Dnca pente costes et de spiritu sancto. Mit Sequenz (AH 53, Nr. 70). – (42r) In festo ss. trinitatis. Mit Sequenz (AH 53, Nr. 81). – (45r) In festo ss. corporis Christi. Mit Sequenz (AH 50, Nr. 385, mit Divisio), Offertorium Sacerdotes mit Prosula (Rubrik: Salutatio, AH 49, Nr. 643). – (50r) In dedicatione ecclesiae. Mit Sequenz (AH 53, Nr. 247). – (52v) Pro defunctis. Mit drei Communiones (zu sätzlich zu Absolve Domine auch CAO Nr. 1420 und Cantus Nr. 204995). 2 (55r–77v) Missae BMV. (55r) De veneracione s. Marie per annum. Mit Kyrie, Gloria mit Tropus Spiritus et alme, Alleluia Ave benedicta Maria mit Tropus (Cantus Nr. 507047), Sequenz (AH 54, Nr. 218), Offertorium Recordare mit Prosula (AH 49, Nr. 624), Communio mit Einschub für De visitatione BMV. – (60v) In adventu domini de s. Maria aut in annunciacione angelica. Mit Sequenz (AH 54, Nr. 191). – (63r) Infra nativitatem et purificationem. Mit Sequenz (AH 54, Nr. 2). – (65r) In pu rificatione BMV: Ante officium canitur hec antiphona (Cantus Nr. 200456). – (65v) Ad missam. Mit Sequenz (AH 53, Nr. 99). – (69v) In annuntiatione BMV. – (69v) Post pasca de s. Maria officium. Mit Sequenz (AH 54, Nr. 18). Mit Hinweis am Schluss: Crist ist erstanden (rot:) Singt das volckh. – (71r) In assumptione BMV. Mit Sequenz (AH 53, Nr. 104). – (73r) In nativitate BMV. Mit Sequenz (AH 53, Nr. 95). – (74v) In visitatione BMV. Mit Sequenz (AH 54, Nr. 193) – (76r) Sequencia de s. Maria per annum communis (AH 54, Nr. 219). Daran anschließend zu sätzliche Sequenz (AH 54, Nr. 245). 3 (77v–101v) Sanctorale. (77v) Iohannes Baptista. Mit Sequenz (AH 53, Nr. 163). – (80r) Petrus et Paulus. Mit Sequenz (AH 53, Nr. 210). – (82r) Paulus. Mit Verweis auf Sequenz (AH 53, Nr. 228). – (83r) Maria Magdalena. Mit Sequenz (AH 50, Nr. 268). – (86r) Laurentius. Mit Sequenz (AH 53, Nr. 173). – (88v) Michael. Mit Sequenz (AH 53, Nr. 192). – (91v) Omnes sancti. Mit Sequenz (AH 53, Nr. 112). – (93r) Martinus. Mit Sequenz (AH 53, Nr. 181 mit Divisio huius propter simplices conf.). – (95v) Catharina. Mit Sequenz (AH 55, Nr. 203). – (97r) Andreas. Mit Sequenz (AH 53, Nr. 122). – (98v) Nicolaus. Mit Sequenz (AH 55, Nr. 265, die letzten zwei Zeilen ohne Nota tion). – (101v) Beginn der Sequenz zu Elisabeth (AH 55, Nr. 120, ohne Notation): Gaude Syon quod egressus a te decor et depressus [ (102r–v) nur Rahmung und Linierung. 4 4.1 (103r–118r) Kyriale. (103r–112r) Kyrie. Gloria. (103r) Summum. Kyrie mit Tropus (AH 47, Nr. 99, 1a, 2a und 3a). – (103r) Aliud summum. Kyrie mit Tropus (AH 47, Nr. 5, 1a, 2a und 3c). – (103v) Angelicum. – (105r) Pascale. – (106v) De apostolis. – (108v) De martiribus. – (109v) De virginibus vel de s. Maria. – (111r) Dominicale. 4.2 (112r–118r) Sanctus. Agnus Dei. (112r) Tit.: Sequitur cantus omnium sanctorum continuus. – (112v) Angelicum. Nur Sanctus. – (112v) Aliud summum. Sanctus mit Tropus (AH 47, Nr. 382). – (113v) Summum. Sanctus. – 354 Nr. 940 (114r) Pascale. – (114v) De dominica nec s. Maria. Sanctus mit Tropus Benedictus Marie filius qui venit … – (115r) De sancta Maria. Sanctus mit Tropus Benedictus Marie filius qui venit … – (115v) Aliud. Sanctus. – (116r) De apostolis. – (116v) De martiribus. – (117r) De confessori bus. – (117v) Dominicaliter. 5 (118r–142r) Commune sanctorum. Jeweils mehrere Introitus, Gradualia, Alleluia, Offertoria und Communiones angeführt. (118r) Tit.: Sequitur commune sanctorum apostolorum. – (118v) [De apostolis]. Mit Sequenz (AH 53, Nr. 228). – (122r) De pluribus martyribus. Mit Sequenz (AH 55, Nr. 14). – (127r) De uno martyre. Mit Sequenz (AH 55, Nr. 9). – (132r) De confessore et pontifice. Mit Sequenz (AH 54, Nr. 88). – (136r) De simplici confessore aut evangelista. Mit Hinweis auf Sequenz (Divisio von AH 53, Nr. 181, Str. 15). – (136r) De virginibus. Introitus mit Angabe von zwei zusätzlichen Versen: 1) De assumpcione beate virginis; 2) Omnium sanctorum. Mit Sequenz (AH 50, Nr. 271) und zwei Offertoria mit unterschiedlicher Melodie (offertorium maius, offertorium minus). 6 (142r–148v) Forts. des Temporale. (142r) Rubrik: Tractus in LXXa continentur folio CLXXII. – (142r) Dnca 1. adventus. – (143v) Dnca 2. adv. – (144v) Dnca 3. adv. – (146r) Dnca 4. adv. – (147r) In cathedra s. Petri. Nur Tractus. – (148r) In quadragesima de uno martire aut confessore. Nur Tractus. (149r–151r) nur Rahmung und Linierung. II (152–229, I) B: Pap. vereinzelt leicht verschmutzt, selten kleine Tintenflecken und Tintenspritzer. Bl. 175 unten leicht ein gerissen. Lagen: (VI–3)160 + 2.V180 + VI192 + 2.V212 + (II+1)217 + VI229 + 1I* (Nachsatzbl., Perg.). Gegenbl. zu Bl. 152, 156 und 160 herausgeschnitten (jeweils ohne Textverlust, trotz Reste urspr. Buchstabenausläufer in Rot auf Gegenfalz zu Bl. 160). Bl. 217 eingehängtes Einzelbl., Falz auf Gegenfalz zum Nachsatzbl. mit Papierstreifen aufgeklebt. In der Mitte der letzten Lage Pergamentfalz. Lagenreklamanten, nur vereinzelt erhalten, vereinzelt auch Seitenreklamanten am Blattrand (vgl. z. B. Bl. 174r, 175r, 193r, 196r). Bl. 171r und 193r am Lagenanfang Kustode XVIII bzw. XX. Am oberen Blattrand zeitgenössische Foliierung in roten römischen Zahlen von I–CXC; Fehler in der Zählung: IX, XI, XII, XXIII, XXXVII, XLVII, LVI, LIX, LXI, LXVIII, LXXI, LXXIII–LXXVIII, LXXX, LXXXIV, CLXXXIV, CLXXXVI und CLXXXVIII ausgelas sen; statt LXVII fälschlich LXXVII; auf LXXXV folgt CLXXVI; LXX zwar nicht angeführt, Blatt in der fortlaufenden Zählung jedoch berücksichtigt (heute Bl. 211). S: Schriftraum 185/205 × 120/135. Von zweifachen roten Tintenlinien gerahmt, zu jeweils sieben Vierlinien systemen samt Textzeile, Bl. 228r–229r ca. 28 Zeilen. Textualis mit kursivem Einschlag, von gleicher Hand wie T. I. N: Gotische Choralnotation auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden, rote vertikale Trennstriche (lange Striche, d. h. über alle vier Notenlinien als Abschluss bei jedem Gesang; kurze Striche zw. den einzel nen Wörtern). A: Rote Paragraphzeichen, Auszeichnungsstriche, Verbindungsstriche (vereinzelt auch in Mintgrün), Trenn striche (bei Notation), Unterstreichungen, Überschriften. Oberlängen und Unterlängen vereinzelt cadellen artig verziert. Rote und tintenfarbene Initialen mit Aussparungen (vereinzelt kopfstempelförmig), Punkt verdickungen und einfachen Ausläufern, bisweilen mit einzelnen Punkten, einander gegenüberstehenden Punkten oder auch Dreipunktmotiv am Buchstabenschaft und vereinzelt auch im Binnenfeld, bisweilen mit Ausläufern in Achterform gelegt. Tintenfarbene Initialen mit roter Tinte umrahmt oder verziert, meist cadellen artig gestaltet. Antiphonarium. 1 (152r–182v) Temporale und Sanctorale, gemischt. (152r) Dnca 1. adventus. – (152v) Dnca 2. adv. – (153r) Dnca 3. adv. – (154r) De s. Maria antiphona. – Dnca 4. adv. – (154v) Rubrik: Hic canuntur O ad singulas dominicas a festo s. Lucie (vier O-Antiphonen). – (155v) In sancta nocte nativitatis dni. Responsorium Iudea et Ierusalem mit Prosula (Hofmann-Brandt Nr. 176). Responsorium Descendit de caelo, Vers Tamquam Nr. 940 355 sponsus mit Prosula (Hofmann-Brandt Nr. 416, CAO Nr. 6411). – (164r) De BMV (gleich wie in Nr. 139 [olim Sign. 14819], hier aber ohne Titel). – (165v) Stephanus. – (166r) BMV. – I ohannes. – (166v) Innocentes. – Dominica infra [octavam]. – In circumcisione dni. – (167r) In epiphania dni. Responsorium Illuminare illuminare Ierusalem mit Prosula (Hofmann-Brandt Nr. 673). – (169r) In conversione s. Pauli. – In purificatione BMV. – (170r) In annuntiatione BMV. – (171r) Alia quere. – In die pasce in matutinas invitatorium (gleich wie in Nr. 139). – (175v) Ad visitationem sepulchri (vgl. Engels 310 [mit Transkription], bezogen auf Nr. 139, Bl. 179r). – (176r) Ad laudes antiphona. Angelus autem domini descendit de celo et accedens … Vorange stellt: (rot:) Wlgus (schwarz:) Crist ist erstanden von der marter allen. – Ad Benedictus. Et valde mane una sabbatorum … – (176v) In vigilia ascensionis. – (177r) In vigilia pentecostes. – (178r) In festo ss. trinitatis: Reimoffizium (LMLO XT31). – (179v) De corpore Christi. Responsorium Homo quidam fecit cenam mit Prosula (Hofmann-Brandt Nr. 448). 2 (182v–185v) Historiae aestivales. (182v) Tit.: Sequuntur historiae estivales. (182v) Et prima Regum. – (183r) Historia Sapiencie. – (183v) Historia Iob. – (184r) Historia Thobie. – (184v) Historia Iudith. – Historia Machabeorum. – (185r) Historia Ezechielis. 3 (185v–193r) Commune sanctorum. (185v) In dedicatione ecclesiae. – (187r) Commune Apostolorum. – (188r) De pluribus martyri bus. – (189v) De uno martyre. – (190v) De confessore et pontifice. – (191v) De simplici confes sore. – (192r) De una virgine. 4 (193r–211r) Forts. des Sanctorale. (193r) Tit.: Sequuntur de sanctis per estatem. – Iohannes Baptista. – (194r) Petrus et Paulus. – (195r) Margaretha: Reimoffizium (AH 28, Nr. 3). Responsorium Quadam die Olibrius mit Pro sula (Hofmann-Brandt Nr. 445). – (196v) Maria Magdalena. Responsorium Accesit ad pedes mit Prosula (Hofmann-Brandt Nr. 696). – (198v) Laurentius. – (199v) In assumptione BMV. – (201v) In nativitate BMV. – (203r) Michael. – (204r) Omnes sancti. – (205v) Martinus. – (207r) Catha rina. Antiphon zum Magnificat (AH 18, Nr. 37 und AH 26, Nr. 69). Responsorium Surge virgo mit Prosula (Hofmann-Brandt Nr. 483). – (208v) Andreas. – (209v) Nicolaus. – (210v) Thomas. 5 (211r–212v) Differentiae et Toni psalmorum, unvollständig. (211r) Tit.: Inductio puerorum ad tonos flectendos in psalmodia. Übersicht (Ed.: E. Vetter, Sum mula tractatus metricus de musica glossis commentarioque instructus [Divitiae musicae artis, A/ VIIIa]. Buren 1988, 89). – (211v) Primus tonus. Secundus tonus. Tertius tonus. – (212r) Quartus tonus. – (212v) Quintus tonus. Sextus tonus. Expl. mut.: … fratres in unum. Magnificat [ (Kustos vorhanden) (213r–227r) nur Rahmung und Linierung. 6 (227v–229r) Dedicatio Ecclesiae (Udalricus?). (227v) Hymnus (AH 51, Nr. 102, mit Divisio). – (228r) Psalmi ad vesperas in solemnitatibus. – (229r) Hinweis in Rot: Capitula et collecte habentur in missalibus. (229v) Hymnus de confessore et pontifice (AH 51, Nr. 118). (Ir–v) leer. 7 (HDS) von späterer Hand ergänzt: Gebete, dt. 1) Pit umb des Nikclas Geczenperger vater sel und umb seiner muter vatter und mutter sel und umb seiner hawsfrau vatter sel et omnium etc. 2) Pitt umb Michel auff Ellen sel und umb seins vater und miter sel und umb seiner ersten hau sfrau vater und muter sel und umb der iczingen vater und muter sel et omnium etc. 356 Neustifter Wappenbuch 3) Pitt umb Michel (gestrichen: auff) Pranttner sel auff dem Kunperch und umb seiner vater und mutter sel und umb seines Iacob veter sel und umb der Pranttnerin vater und muter sel und antecessores et omnium fidelium. G. G. / U. S. ohne Sign. NEUSTIFTER WAPPENBUCH Perg. I, 88, I Bl. 245 × 180. Sterzing (?), 1548–1552. B: Lagen: (I–1)I (Vorsatzbl., Pap. 18. Jh.) + 16.II124(62) + (II–1)130(65) + 2.II146(73) + I150(75) + (II–1)156(78) + 2.II172(86) + (II–2)176(88) + (I–1)I* (Nachsatzbl., Pap. 18. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 65 (S. 129/130) und 72 (S. 153/154) herausgeschnitten (Textverlust?). Am oberen Blattrand Paginierung, beginnend mit der Zahl 2, Anfang 20. Jh. radiert und ersetzt, beginnend mit der Zahl 1 (wohl durch Marie Fischnaler, s. Arch 11). Eingelegtes Bl. (gefaltet) mit zwei schwarzweiß ausgeführten und einem kolorierten Wappen. S. 53–77 kleine Papiermarken mit Buchstaben „M“ (S. 55, zweite Zeile Mitte „55 M“) in Tinte über den Wappen aufgeklebt, nachträglich. Bisweilen über den Wappen Klebespuren derartiger Papiermarken sowie Reste von rotem, vereinzelt auch grünem Wachs (?), das als Klebstoff verwendet wurde. S: Zuweisungen über dem jeweiligen Wappen von unterschiedlichen Händen, Haupthand bastardaähnliche Buchschrift, wohl Vigil Raber. A: Da das Wappenbuch durch die Edition von H. W. Arch umfassend und rezent erschlossen ist, an dieser Stelle nur grundlegende Angaben zur Wappenanzahl, Technik, Händescheidung und Datierung. Insgesamt 1562 Wappen (koloriert und unkoloriert, mit und ohne Namen der Wappenträger; ohne Leerwap pen [13] und Hauszeichen [2]); Zahlenangabe 1558 (S. 176) evtl. keine Jahreszahl, was aber nicht völlig verworfen werden sollte, sondern Wappenzählung (s. Arch 16): 1562 minus vier übermalte, unfertige oder verschmierte Wappen; in der Regel drei mal drei Wappen pro Seite, S. 176 nur zwei Wappen. Rundbogige Schilde (54 × 47, mit Schablone angelegt). S. 104–105 Umrisszeichnung mit schwarzer Tinte, 104,7 zusätzlich Vorzeichnung erkennbar (nur 105,8 koloriert); ansonsten Ausmalung mit Deckfarben (ohne Verwendung von Blattgold), fallweise dünnerer Farbauftrag; Originalkolorit etwas kräftiger als in Ed. Seiten verkehrte Darstellung unsymmetrischer Wappen (möglicherweise für geplanten Druck, s. Arch 14–15). Plastizität von Holz- bzw. Steinschilden (mit Graten) wird durch verschattete Kanten und horizontal gestri chelte (mit Pinsel ausgeführte) Schattenpartien in Grau bzw. in dunklerer Lokalfarbe erzeugt; auch Wappen bilder durch Schattenwürfe immer wieder räumlich hervorgehoben (z. B. 7,4). Es fällt auf, dass ab S. 106, also nach dem Abschnitt mit den unkolorierten Zeichnungen, bis einschließlich S. 116 Binnenzeichnungen minutiöser ausgeführt sind, vgl. z. B. die einfache Fellstrukturierung des Löwen 85,2 mit den bewegten Binnenmodellierungen S. 107; zudem verstärkte, die Plastizität von Schilden und Motiven erhöhende Flächenschattierung: hier wohl andere Hand tätig; ab S. 117 bis S. 123 möglicherweise wieder die urspr. Hand tätig; danach vergröbert sich die Ausführung bis zum Ende der Hs. Die beiden Wap pen S. 176, v. a. 176,2, könnten Ergänzungen im Stile der voranstehenden Wappen sein. Inwieweit daraus (in Zusammensicht mit Handwechseln in der Schrift) de facto auf Handwechsel in der malerischen Ausgestal tung geschlossen werden kann, die in der Hauptsache sicherlich Raber zugeschrieben werden muss, bleibt noch zu untersuchen. Arch gibt das Wappenbuch in die letzten Lebensjahren Rabers (vor 1552, s. auch bei G). Sollte es sich bei den Wappen S. 176 tatsächlich um Ergänzungen handeln, dann könnte 1558 doch eine Jahreszahl sein, die auf die Ergänzungen Bezug nimmt. S. R. E: Neuzeitlicher Bibliothekseinband: schwarz gesprenkeltes braunes Pap. über Pappe. Neustift (?), 2. H. 18. Jh. Pap. tw. stark abgerieben. Am VD und HD Ecken mit braunem Leder verstärkt. Rücken: braunes Leder, fünf einfache Bünde, von Goldbordüren gesäumt. In den Feldern 1 sowie 3–6 in Goldprägung Einzelstempel kleine Blumensträußchen (Vignetten) frei (Nr. 1). Zw. erstem und zweitem Bund rotes Titelschild mit Gold druck WAPPEN[BU]CH. Schnitt rot gefärbt. Spiegel Pap. 18. Jh. Am Spiegel des VD Exlibris, Besitz vermerke und Signaturen (s. bei G), Spiegel des HD leer. Graduale Neocellense I 357 G: Entstanden zw. Vigil Rabers Arlberg-Aufenthalt 1548 und dessen Tod 1552. Am Spiegel des VD alte Signa turen, wohl nicht der Stiftsbibl. Neustift: 2602 (Tinte), Z. I. c. C. (Tinte, gestrichen) und Z. III. l (Bleistift). Besitzvermerk Stiftsbibl. Neustift 18. Jh. Bibliothecæ Neocell., mit Exlibris der Stiftsbibl. Neustift 18. Jh. überklebt. Darunter in Bleistift (fälschlich) 1.576 Wappen. S. 176r 1558 (Tinte), am Spiegel des VD 155(9) (Bleistift, schlecht lesbar), wohl nicht Jahreszahl, sondern Wappenzählung (nicht korrekt). Bei der vorüber gehenden Klosteraufhebung 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. L: C. Fischnaler, Vigil Rabers Wappenbuch der Arlberg-Bruderschaft in Weimar. Der deutsche Herold. Zeit schrift für Wappen-, Siegel- u. Familienkunde 14 (1909), Nr. 8, 153–176, hier 162–162. – G. Gerola, Il codice araldico di Novacella in Alto Adige. Bollettino ufficiale della consulta araldica del Regno 9, 43 (1937) 127–164. – Rasmo 63 (Nr. 235) und Abb. 189. – H. W. Arch, Vigil Rabers Neustifter Wappenbuch. Aus der Bibliothek des Augustiner Chorherren-Stiftes Neustift. Brixen 2001. (Ir–v) leer. Vigil Raber: Neustifter Wappenbuch (Ed. und Faksimile mit Blasonierungen sowie ausführlicher Ein leitung: H. W. Arch, Vigil Rabers Neustifter Wappenbuch. Aus der Bibliothek des Augustiner Chorher ren-Stiftes Neustift. Brixen 2001). Insgesamt 1577 Wappendarstellungen, darunter zwei Hauszeichen und 13 leere Wappenschablonen. 352 ohne Namenszuweisungen. Wahrscheinlich diente das Wappenbuch des Jörg Rugen(n), Innsbruck, ULB Tirol, Cod. 545 im Origi nal oder in einer Kopie als Vorlage, vgl. bes. Bl. 249r–260r „Vnder dem keissertom Constantinoppel“ (Phantasiewappen). Vgl. Weimar, Herzogin-Anna-Amalia-Bibl., Fol. 220: Vigil Raber, Wappenbuch 1548 (Konrad Fisch naler, Vigil Rabers Wappenbuch der Arlberg-Bruderschaft in Weimar. Der Deutsche Herold 8 [1909] 1–24). Eingelegter Zettel mit drei Wappendarstellungen (jeweils mit Helmzier). (Ir–v) leer. C. S. ohne Sign. GRADUALE NEOCELLENSE I Perg. II, 289, I Bl. 725 × 505. Neustift, 1442. B: Hs. um 1980 restauriert (s. bei G). Perg. am rechten unteren Blattrand tw. stark abgegriffen, daher Ecken häufig im Zuge der Restaurierung ergänzt bzw. ersetzt. Perg. tw. dünn und Tinte durchscheinend. Einzelne Blätter oder Teile von Blättern stark vergilbt (insbes. Bl. 31r–v unten, 74v, 76v, 156v, 209r–v, 274r–v, 277r– v). Am Blattrand vereinzelt kleine Löcher und Risse, Nahtspuren. Risse und Fehlstellen tw. mit Pergament geklebt. Zahlreiche Bl. verloren gegangen (Textverlust, s. bei Inhalt), Gegenfälze der Einzelbl. meist im Zuge der Restaurierung verstärkt bzw. erneuert. Bisweilen Korrektur durch Aufkleben von zusätzlichen Pergamentstreifen und -stücken (z. B. Bl. 199v, 213r, 214r, 215v, 245r). Zahlreiche Melismen zur Verein fachung und Modifizierung der Melodien mit Papierstreifen mit rotem Vierliniensystem (meist leer, vereinzelt Kustode neu aufgetragen) im 16./17. Jh. überklebt (Bl. 1v, 2r, 3v, 4r, 4v, 6v, 8v, 17r, 17v, 19r, 48v, 49r, 51r, 51v, 52r, 54r, 57v, 59r, 62v, 63r, 70r, 76r, 76v, 88r, 88v, 89r, 116v, 117r, 118r, 130v, 131r, 132r, 136v, 139v, 141r, 142r, 282v, 283r), heute jeweils nur mehr am linken Rand festgeklebt, darunter Klebespuren und Durchstreichungen sowie Bleistifthinweise und tw. Abgrenzungen in Rot deutlich sichtbar. Papierstreifen vereinzelt verloren gegangen (vgl. Klebespuren bzw. Verfärbungen). Lagen: (I–1)I (Vorsatzbl. 20. Jh.) + 1II + (IV–1)7 + 3.IV31 + (IV–1)38 + (IV–2)47(44) + (IV–1)55(51) + 3.IV79(75) + (IV–6)87(77) + IV95(85) + (IV–1)103(92) + 3.IV127(116) + (IV–2)135(122) + IV143(130) + (IV–8)151(130) + (IV–1)158(137) + (IV–3)167(142) + 2.IV183(158) + (IV+1)191(167) + IV199(175) + (IV–6)207(177) + IV215(185) + (IV–2)223(191) + 3.IV248(215) + (IV–1)255(222) + IV263(230) + (IV–3)271(235) + IV279(243) + (IV+1)287(252) + IV295(260) + (IV+1)303(269) + IV311(277) + (IV+4)325(289) + (I–1)I* (Nachsatzbl. 20. Jh.). Urspr. Bindung bzw. Lagenordnung bei Restaurierung tw. nicht berücksichtigt (z. B. Bl. 218 und 220 [urspr. Gegenbl. zu Bl. 219 und 221] herausgeschnitten, Gegenfälze heute zw. Bl. 217 und 219 und somit Bl. 219 fälschlich der zweiten Lagenhälfte zugeordnet). 358 Graduale Neocellense I Bl. II eingehängtes Einzelbl., anderes Perg. kleineren Formats (690/705 × 450/460), urspr. wohl als Spiegel auf dem VD aufgeklebt (Holzfaserung auf Recto sichtbar), heute gelöst und auf neuen Pergamentfalz aufge klebt (zw. Bl. 6 und 7). Bl. 7 eingehängtes Einzelbl., möglicherweise urspr. Gegenbl. zu heutigem Bl. II., Gegenfalz nach Lage 2 (zw. Bl. 15 und 16). Gegenbl. zu Bl. 32, 45, 46, 50, 102, 134, 135, 152, 165, 166, 167, 219, 221, 255, 270 (Textverlust), Gegenbl. zu Bl. 218 und 219 (ohne Textverlust) herausgeschnitten. Zusätzlich Bl. 81–86, 144–151 (= Lage 19), 201–206, 266/269, 322 und 323 (Textverlust) herausgeschnitten. Bl. 318–321 eingehängte Einzelbl. Zw. Bl. 187 und 188, 282 und 283, 301 und 302 jeweils ein später ein gehängtes Einzelbl., Perg., in zeitgenössischer Foliierung nicht beachtet, mit Bleistift als „187bis“, „282bis“ bzw. „301bis“ foliiert. Am Lagenende jeweils Kustoden in römischen Zahlen. Lagenreklamanten. Am oberen Blattrand zeitgenössische Foliierung in roten römischen Zahlen I–CCCXXIIII; Fehler in der Zählung: 244 ausgelassen. Bl. 28, 36, 157, 199 und 227 Spuren von Signakeln. S: Schriftraum 535/540 × 350/360, seitlich von zweifachen roten Tintenlinien begrenzt (linke Spalte Noten schlüssel, rechte Spalte Kustos), jeweils sieben rote Vierliniensysteme (39 mm) mit Textzeilen. Bl. 325r–v: zwölf bzw. acht Zeilen auf roter Tintenlinierung. Zirkelstiche. Textualis formata (Textura), dat. 7. Septem ber 1442 (Bl. 325v). Schreiber: Friedrich Zollner, Chorherr aus Langenzenn (Bl. 325r; zu Friedrich Zollner s. Giner 10: gest. 1449; laut Eintrag auf Bl. 325v gest. 1446). Nur vereinzelt Radierungen und Korrekturen (sowohl bei Noten als auch bei Text). Zahlreiche Radierungen, Durchstreichungen, Ergänzungen und Korrekturen von späteren Händen (Bl. 324r: dat. 1512), meist ebenfalls in Textualis formata (Textura), tw. auch in Rotunda (z. B. 271r, Z. 2–7). Blatt gestaltung (Höhe der Vierliniensysteme sowie Schriftgröße) meist nachgeahmt, vereinzelt Abweichungen (z. B. Bl. 287bisr nur sechs Notensysteme). Zusätze in schwarzer und roter Tinte sowie Bleistift bis ins 18. Jh., z. B. Vacat, Gliederungsstriche von unterschiedlicher Breite, Rubriken, Buchstaben für die Aufführungs praxis (mit Orgel? bzw. Bl. 223r–v, 225r in tw. pastosem Rot Chorus – Rectores), Blattverweise. Mehrere Feste zur Gänze von späterer Hand gestrichen. N: Gotische Choralnotation (mit Virga) auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden. Kustoden in Rot weisen auf einen Wechsel des Schlüssels innerhalb eines Gesangs hin. Reperkussionston mit einem orna mentalen Haarstrich. Bl. 297v und 306v (Tropus zum Sanctus bzw. Agnus Dei) metrische Notation mit Zeichen der Mensuralnotation. Bl. 301bisr–v von anderer Hand 15. Jh.: sog. „umgekehrte Hufnagelnotation“ (vgl. hierzu auch Graduale II). Im 16./17. Jh. zahlreiche Melismen gestrichen und durch Pergamentstreifen überklebt, Notation bisweilen ergänzt bzw. verändert. Hinzufügung von zahlreichen, tw. sehr dicken schwarzen vertikalen Gliederungsstrichen am Beginn und am Schluss der zu streichenden Melismen (z. B. Bl. 4v). Im 18. Jh. Ergänzung von langen Gliederungsstrichen in Rot (oben lilienförmig bzw. Rhombus form), die über das Vierliniensystem hinausreichen (z. B. Bl. 1r), möglicherweise Hinweis auf AlternatimAufführungsform, vielfach in Verbindung mit den Rubriken Rectores und Chorus (z. B. Bl. 110r). Bl. 271v– 310v (Kyriale) am Blattrand entsprechend den verschiedenen Abschnitten Ergänzung der Buchstaben A, B, C, D: Bedeutung nicht eindeutig geklärt, möglicherweise jedoch Hinweis auf Aufführungspraxis. Von noch späterer Hand zahlreiche Vorzeichen (# und b) in Schwarz und Rot ergänzt. A: Rubriken und Foliierung in Rot; spätere Zeigehand Bl. 4r. Fleuronnée: gut 800 Fleuronnée-Initialen (abwechselnd rote und blaue, in der Regel eine Notenzeile hohe Buchstaben mit Punktverdickungen und wenigen herzblattförmigen Verzierungen, z. B. Bl. 150v; in der Regel keine Aussparungen, die Palmettenaussparungen Bl. 136r wohl von anderer Hand; Einleitungsinitiale des Sanctus-Teils Bl. 291r in der Höhe von zwei Notenzeilen mit rot-blauem, durch schraffierte Blattaus sparungen verziertem Buchstabenkörper; Bl. 319v und 321r zwei rot-blau kopfstempel‑ bzw. bogenförmig gespaltene Initialen von späterer Hand) und ca. 400 fleuronnierte Cadellen, zumeist in der Höhe einer No tenzeile; von diesen etwa 1200 erhaltenen fleuronnierten Buchstaben die allermeisten vom Hauptflorator; nur wenige Fleuronnée-Nachträge (von mindestens acht Händen, s. u.). Das Fleuronnée der Haupthand ungeachtet der großen Anzahl der zu dekorierenden Buchstaben sehr gleichmäßig und akkurat ausgeführt; wenige Motive, die trotz unterschiedlichster Kombinationen einen relativ gleichförmigen Gesamteindruck erzeugen. Außer bei Cadellen in den obersten Notenzeilen, die aufwändiger geschmückt sein können (z. B. Bl. 23v, 91r, 118v, 305v), das Formenvokabular des Cadellen-Fleuronnée sehr ähnlich, aber noch etwas re duzierter (z. B. weniger Besatz); Cadellen-Fleuronnée in Schwarz, mit wenig rotem Dekor, vorwiegend Konturlinien und Punkte. Die blauen Initialen mit Fleuronnée in tw. changierenden Rot‑ und Violetttönen (Hell‑ und Dunkelrot, Orangerot bis Bräunlich, oft sehr blasses Violett, Rosa), seltener in Grün; rote Initialen vorwiegend mit grünem, seltener mit rotem oder violettem Fleuronnée; fallweise Farbkombinationen im Fleuronnée, z. B. Bl. 195r (grün-violett) und 291r (grün-rot), ab und an Profilköpfe (s. u.) in Rot zusammen mit andersfarbigem Ornament, z. B. Bl. 112v (grün), 215v2 (violett), 270v (grün-violett); wenige Fälle von andersfarbigen, evtl. nachgetragenen Punkten und/oder Kernen (z. B. Bl. 25r4). Graduale Neocellense I 359 Drei Knospenformen: (1) spitz, auch zu Paaren bzw. Ähren zusammengefasst (z. B. Bl. 249r1) oder als Knospenwiegen (häufig als Binnenfeldgliederung, s. u.); (2) rund, zumeist stark eingerollt, auch als Ähren (z. B. Bl. 7v1, oberes Binnenfeld) oder Knospenwiegen (z. B. beim „bedachenden“ Besatzmotiv, s. u.); (3) rund und durch Doppelstriche mit den Stielen der Knospenwiegen verbunden (z. B. als Besatz der Initiale Bl. 55v); (2) und (3) auch im Wechsel mit Dornen. Gestrichelte Halbpalmetten, die den spitzen Knospen ähneln, aber größer sind als diese und sich eng an ihre Stiele zurückklappen (z. B. Bl. 212r). Binnenfelder immer wieder in Kompartimente, oftmals Streifen (v. a. friesartig übereinander gesetzte Knospenwiegen, z. B. bei der Initiale Bl. 102r) unterteilt, bisweilen Kachelungen (so ergeben aneinander gesetzte Dreiecks formen, gefüllt mit Knospenpaaren, wie (1), Herzformen, z. B. Bl. 105v1 und im gesamten großflächigen, teppichartigen Initialfeld Bl. 291r; Bl. 208v2 quadratische Kachelung, mit Kreuzverbindungen), Medaillons bzw. S-förmige Doppelspiralen (z. B. Bl. 107v); aus vollfarbigem Binnenfeldgrund ausgespart werden u. a. die gekachelten Knospenpaare und die zu Ähren zusammengefassten Halbpalmetten (z. B. Bl. 212r) oder Doppelknospen, die runde Knospen und Halbpalmetten kombinieren können (z. B. in den Binnenfeldern der Initiale Bl. 78v); speziell in den Binnenfeldern von Cadellen aus schwarzem Grund ausgesparte ondulieren de Halbpalmettenranken (z. B. Bl. 17r) und Halbblätter mit sanft eingebuchteten und in regelmäßigen Ab ständen tief eingeschnittenen Rändern, am Ende des Einschnitts jeweils ein kleines „Loch“ (z. B. bei der oberen Cadelle Bl. 163v; zur Herleitung dieses Motivs als Negativform von Kopfstempeln s. S. Rischpler, Der Illuminator Michael. Purkersdorf 2009 [Codices Manuscripti. Suppl. 1], 49). Konturlinien. Als Besatz immer wieder runde Knospen, u. a. weit auseinander stehend (z. B. Bl. 208v2), auch als gegeneinander ge kehrte Paare (z. B. unten an der Initiale Bl. 114v) oder seltener als Ähren (z. B. Bl. 126r); ab und an Laufen der Hund (z. B. Bl. 246v) sowie ein dreieckiges Motiv, das häufig bedachend eingesetzt wird: auf einer Knospenwiege, hier mit runden Knospen, sitzt eine gefüllte Dreiecksform, deren Schrägen von Fadenaus läufern begleitet werden, die von den Knospen ausgehen und mit den Schrägen durch Doppellinien ver spannt sind (z. B. Bl. 12r); sehr oft Halbpalmetten, die durch Zackenlinien verbunden sind, wodurch sich charakteristische Ketten aus rautenartigen Gliedern ergeben, welche zusätzlich mit Doppelstrichen verziert sind (z. B. Bl. 7v1); nur bei der aufwändigsten Initiale dieses Gradualbandes, Bl. 291r, Besatzmotiv aus ge reihten, mit den Spitzen zueinander zeigenden Halbpalmettenpaaren, zw. denen jeweils eine eng an ihren Stiel geklappte Halbpalmette aufragt; über den Palmettenpaaren zwei einander zugewandte Elemente des Laufenden Hunds mit einem hochstehenden Herzblatt in der Mitte. Charakteristisches Besatzmotiv: Profil köpfe (maximal fünf Köpfe an einer Initiale, Bl. 270v; Bl. 177r in der untersten Zeile zum urspr. I mit Be satzkopf weiteres I spiegelsymmetrisch ergänzt, um den Kopf in ein Binnenfeld zu integrieren), die gegen ständig (z. B. Bl. 52v1), spiegelsymmetrisch über- (z. B. Bl. 246v) oder wie Janusköpfe nebeneinander angeordnet sein können (z. B. an beiden Cadellen auf Bl. 143r); die zumeist langgezogenen Köpfe eher stereotyp gestaltet: markanter Wulst über der Nase, der die niedrige Stirnpartie stark gerunzelt wirken lässt und in Kombination mit dem schmal geschnittenen, oft etwas schräg gestellten Auge (stets mit geschwun genem Brauen‑ und betonendem Unterstrich) und der dünnen, meist deutlich nach unten gezogenen Mund linie einen ernsten, fallweise mürrischen Eindruck erzeugt; bei Cadellen zuweilen rote Zunge ergänzt (z. B. Bl. 305v1) oder Bl. 2v1 ein Blättchen. Variationen eigentlich nur bei den Nasenformen (z. B. Bl. 31v1 sehr ausgeprägte Nase) und in der Gestaltung von Haar und Kopfbedeckung: bei Profilköpfen der FleuronnéeInitialen höhere „Stoppelfrisuren“ oder dünnere Haarkränze (Tonsuren?, s. z. B. Bl. 178r) aus feinen Parallel strichen (der Bart Bl. 78v dürfte ergänzt sein); Cadellen-Köpfe tragen häufig mehr oder weniger hohe kegel förmige Mützen mit breiten Krempen. Konturlinien. Nicht allzu lange, oft doppelgestrichelte und Achterschlaufen bildende Fadenausläufer, tw. auch um Ausläufer der Initialen gewickelt, selten gebündelt (z. B. Bl. 163r: rote Bündelstriche, Fäden doppelgestrichelt, symmetrisch angeordnet und mit zurückschwing enden Häkchenenden); Fibrillen; nur wenige Fleuronnée-Leisten, jeweils mit Fadenausläuferbündeln an den Enden: Bl. 271v u. a. aus spiegelsymmetrisch zueinander gebogten Knospenähren; Bl. 291r Leisten, bei denen der charakteristische, mit Herzblättern garnierte Besatz (s. o.) spiegelsymmetrisch gedoppelt wurde. – Bl. 33r und 36v nehmen die in Deckfarbe ausgeführten Randdrolerien (s. u.) eindeutig Bezug auf die Fleu ronnée-Initialen bzw. orientieren sich auf diese hin, woraus für den Arbeitsablauf geschlossen werden kann, dass die Fleuronnée-Initialen zusammen mit Notation und Texten vor der Deckfarbenausstattung fertig gestellt waren. Der Hauptflorator ist darüber hinaus auf dem Einband von Cod. 540 (Cadelle mit Profilköpfen) sowie in den Handschriften Cod. 76 und Brixen, Priesterseminarbibl., Cod. C.2 (jeweils eine Fleuronnée-Initiale) nach weisbar – allesamt Zeugnisse ohne Deckfarbenschmuck, die im 5. Jz. des 15. Jh. in Neustift oder Brixen entstanden sind. Er ist aber auch in drei älteren Hs. (ebenfalls ohne Deckfarbenschmuck) belegt, die im Augustiner Chorherrenstift Neunkirchen am Brand bei Nürnberg geschaffen wurden (Bamberg, Staatsbibl., Msc. Patr. 40 [1415], Msc. Patr. 164 [um 1415] und Msc. Patr. 147 [1420]; s. Die Handschriften und Drucke des 15. und frühen 16. Jahrhunderts der Staatsbibliothek Bamberg. Katalog der illuminierten Handschriften der Staatsbibliothek Bamberg, Bd. 4, in Vorbereitung). Dieser Buchmaler, von dem aus der geraumen S panne zwischen seiner Tätigkeit in Franken und seinem Wirken in Südtirol (bislang) keine weiteren Zeugnisse 360 Graduale Neocellense I bekannt sind, könnte gemeinsam mit dem aus Langenzenn, dem Tochterstift von Neunkirchen am Brand, stammenden Schreiber Friedrich Zollner oder auf dessen Anregung nach Neustift gekommen sein, wohin er (ost-)mitteleuropäisches Formenvokabular mitbrachte. So sind seine Profilköpfe in der mitteleuropäischen Buchmalerei des 14./15. Jh. weit verbreitet und begegnen beispielsweise in der großen Neunkirchener Bibel von 1389/90 (Bamberg, Staatsbibl., Msc. Bibl. 9/1–3, hier im Motivschatz der 2. Hand, z. B. Msc. Bibl. 9/1, Bl. 1v; s. K.-G. Pfändtner, S. Westphal, Die Handschriften des 14. und 15. Jahrhunderts der Staatsbibliothek Bamberg. Katalog der illuminierten Handschriften der Staatsbibliothek Bamberg, Bd. 3. Wiesbaden 2015, 74–79, Kat.-Nr. 42, Abb. 154; mit der zweiten Hand der Neunkirchener Bibel arbeitete der Hauptflorator in Msc. Patr. 164 [jeder eine Fleuronnée-Initiale] und Msc. Patr.147 [an einer Fleuronnée-Initiale] zusammen). Aus der ostmitteleuropäischen Buchmalerei entlehnte der Hauptflorator z. B. die Buchstabenkörper mit aus gesparten Blattformationen oder die sich eng an ihre Stiele zurückklappenden Halbpalmetten, vgl. z. B. Stuttgart, Württembergische LB, Cod. poet. et phil. 2o 34, Bl. 1r, 79r (Böhmen / Schlesien, 2. Viertel 14. Jh., s. P. Burkhart, Die gotischen Handschriften der Württembergischen Landesbibliothek Stuttgart, Bd. 2: Vom späten 13. bis zum frühen 15. Jahrhundert. Wiesbaden 2005, 109–110, Kat.-Nr. 69, Abb. 250, 253; Volldigi talisat der Hs. unter digital.wlb-stuttgart.de/purl/bsz408922788). Für die großformatigen Gradualbände mit ihren annähernd 3000 Initialen und Cadellen musste der Hauptflorator seine Gestaltungskonzepte adaptie ren, wobei auch italienischer Einfluss vorbildgebend gewesen sein könnte, vgl. die opulenten Initialen Bd. 1, Bl. 291r und Bd. 2, Bl. 254r, 278r, deren Binnenfeldgründe farbig ausgemalt sind, mit Vorbildern aus der Toskana, wo im 14. Jh. Chorbücher mit großformatigen Fleuronnée-Initialen geschaffen wurden, die man ab Ende des 14. Jh. mit üppigen, seitenumrahmenden Ausläufern versah (s. RDK, Fleuronné, Sp. 1180) – als Beispiel hierzu ein Hymnar aus dem frühen 15. Jh.: London, British Library, Add. 30014, Bl. 16r; s. G. Vailati Schoenburg Waldenburg, Un innario senese del 1415, in: E. Sesti (Hrsg.), La miniatura italiana tra Gotico e Rinascimento. Firenze 1985, Bd. 2, 545–562, Fig. 6. Auch die erste Deckfarbenhand führte etwas Fleuronnée aus (zu diesem Miniator s. beim zweiten Gradual band unter „Deckfarbenschmuck“ sowie Rischpler): z. B. Bl. 36v Ähren aus spitzen, gepunkteten Knospen auf die blattvergoldeten Außengrundzwickel der Deckfarbeninitiale mit weißlichem Malmittel aufgetragen, vgl. goldenes Binnenfeld-Fleuronnée auf Bl. 175r des zweiten Gradualbandes, s. des Weiteren Cod. F.8 der Brixner Priesterseminarbibl., dessen Deckfarben‑ und Fleuronnée-Schmuck (Goldfleuronnée in Binnen feldern von Deckfarbeninitialen sowie rotes und blaues Lombarden-Fleuronnée) fast zur Gänze von dieser Hand stammt, oder die Gutenberg-Bibel München, BSB, 2 Inc. s.a. 197 (gedruckt Mainz, 1454/55, nicht nach 24.8. bzw. 15.8.1456), s. beispielsweise Fleuronnée-Initiale im ersten Bibelband, Bl. 1r (http://daten. digitale-sammlungen.de/bsb00004647/image_5). Zudem mindestens acht (jüngere) Fleuronnée-Hände, großteils unter Einfluss des Hauptflorators: (1) Bl. 316v–319v: an 23 Initialen und Cadellen variationsreiches, sauber gezeichnetes Fleuronnée, das wie (2) plastisch herausmodellierte Blattranken als Binnenfeldfüllungen zeigt (z. B. Bl. 317v3); diese Hand auch im zweiten Gradualband tätig (Ergänzungshand 1). (2) Bl. 187bisr–v und 321r: Fleuronnée von der geübten, wohl in das 1. Fünftel 16. Jh. zu datierenden Hand, die in Cod. 405 (dat. 1501, hier das Fleuronnée flüchti ger), Cod. 194 (dat. 1507), Cod. 142 (Teil mit in Rede stehendem Fleuronnée, dat. 1521) zu finden ist, eine Reihe von Initialen im zweiten Band des Zollner-Graduales gezeichnet und Sign. 14820 zur Gänze ausge stattet hat (ausführlich zu dieser Hand bei Sign. 14820); bei den vier Initialen Bl. 187bisv orientierte sie sich stark an der größten Fleuronnée-Initiale, die der Hauptflorator im ersten Gradualband geschaffen hat (Bl. 291r), vgl. z. B. den charakteristischen, mit Herzblättern garnierten Besatz; es ist eher unwahrscheinlich, dass diese Hand auch die aufwändige Cadelle Bl. 187bisr (u. a. zwei kleine Drachen mit verschlungenen Hälsen als Besatz) gestaltet hat. (3) Bl. 273v4, 278v5, 314v: unsicher fleuronnierte Initialen, Bl. 278v5 mit „Janusköpfen“. (4) Bl. 91v, 245r (hier auf eingeklebtem Streifen mit schwungvollem Profilkopf): flüchtig dekorierte Cadellen. (5) Bl. 14r: fleuronnierte Cadelle (zusammen mit ergänzter Notenzeile). (6) Bl. 99r: an einer Initiale Besatzkopf radiert, schwarzes Fleuronnée des Hauptflorators belassen und rotes, ungelenkes Fleuronnée ergänzt. (7) Bl. 324r (mit Datierung 1519): größere Fleuronnée-Cadelle von einer Hand, die wie derum (2) nachahmt. (8) Bl. 325r (Kolophon): schwarzer Fleuronnée-Dekor (mit betonender Funktion) in den Binnenfeldern des N von Nicolaus und des Q von Qui incepit, von der Hand, die den auf Propst Nikolaus Scheiber bezogenen Sterbevermerk geschrieben hat. Frontispiz. Bl. IIv (eingehängtes Pergamentblatt, s. bei B): Thronende Madonna in lavierter und partiell kolorierter Federzeichnung (in Hell‑ und Dunkelbraun). Kopie nach dem wohl urspr. Frontispiz des Gradual bandes, das 1925 aus einer unbekannten Berliner Sammlung vom Städel (Frankfurt a. M.) erworben wurde, wo es sich noch heute befindet (Inv.-Nr. SG 355). Bis auf die Maße (Original 710 × 485, leicht unregelmäßig; Kopie 690/704 × 450/460) äußerst getreue Kopie, die beispielsweise den beschnittenen oberen Rand der Zeichnung sowie deren delikate Farbgebung minutiös berücksichtigt, aber den Abklatsch der Rubrik auf Bl. 1r des ersten Gradualbandes ignoriert, der auf dem Original sicht‑ und entzifferbar ist; zudem differieren die Physiognomien der Madonnenfiguren minimal (während die kopierte Muttergottes ernst blickt, scheint sie auf dem Original zu lächeln). Gerade die Beschneidung des Blattes lässt daran zweifeln, ob die Frankfurter Graduale Neocellense I 361 Zeichnung explizit für das Graduale geschaffen wurde (vgl. Roland, Gotik 287–288 [Kat. 184], hier 287; mit Abb., die allerdings die Kopie, ohne Schriftabklatsch, zeigt); eine sekundäre Verwendung ist wahrschein licher, so schlagen Brinkmann–Kemperdick (s. u.) vor, dass das originale Frontispiz als Vorzeichnung für eine Wandmalerei angelegt worden sei. – In imposanter und mittels Fialen, Maßwerk, bewegten Kreuz blumen, Akanthusblatt‑ und Krabbenbesatz detailreich gestalteter Thronarchitektur mit Podest, apsisartig gerundeter Sitznische und einem an ein Altargesprenge erinnernden Aufbau wird die auf einer Bank sitzende und ihr Kind zärtlich umfassende Gottesmutter von zwei Kerzen tragenden Engeln mit heraldisch anmuten den Flügeln flankiert; Mantel der Gottesmutter reich gefältelt, auffällig ihre große, üppig mit Perlen und Edelsteinen geschmückte Krone, die abgeflacht wirkt; im Giebel über Thronnische, von einem Strahlen kranz umgeben, Kopf Gottvaters, der seine Rechte zum Segen erhebt und mit der Linken die Taube des Heiligen Geistes präsentiert; darunter zwei Posaunenengel, von deren Instrumenten weiße Fahnen mit roten Tatzenkreuzen abflattern; zwei Seitengiebel: links Verkündigungsengel, rechts Maria. S. auch ausführliche Beschreibung mit Abb. bei B. Brinkmann, St. Kemperdick, Deutsche Gemälde im Städel 1300–1500. Mainz 2002, 155–161 sowie https://sammlung.staedelmuseum.de/de/werk/thronende-madonna-mit-engeln (mit Abb.); detaillierte Beschreibung ebenso bei Hermann 221–222. – Es ist anzunehmen, dass das originale, jetzt in Frankfurt aufbewahrte Blatt nicht allzu lang vor der Fertigstellung des ersten Gradualbandes 1442 geschaffen wurde, wahrscheinlich von einem (Süd‑)Tiroler Zeichner, der definitiv unter (ober‑)italienischem Einfluss stand (s. Hermann 221–222; Brinkmann–Kemperdick; Roland, Gotik 288); die Kopie dürfte an lässlich der Veräußerung des Originalfrontispizes Anfang 20. Jh. entstanden sein. Deckfarbenschmuck: ein Wappen; acht (erhaltene) figürliche Initialen mit bevölkerten Ranken (Bl. 1r, 27v, 33r, 36v, 156v, 200r, 227v, 261r; bis auf das fünfzeilige, fast 400 mm hohe P auf Bl. 27v Initialen in der Höhe von zwei bis drei Notenzeilen) von zwei Händen: Bl. 1r von der ersten, der Rest von der zweiten Hand. Darüber hinaus mindestens zwei Folios mit figürlichem Deckfarbenschmuck herausgetrennt; diese Seiten (Bl. 129 und 218: Dnca in palmis und In festo ss. trinitatis), die bei Laußermayer nicht mehr aufgeführt sind, von Hermann noch beschrieben, was bedeutet, dass sie zw. 1905 und 1965 entfernt worden sein müssen. Zu Einordnung und Charakteristika der ersten Hand s. beim zweiten Gradualband unter „Deckfarben schmuck“, der zur Gänze von diesem Illuminator stammt, sowie Rischpler; nachfolgend Einordnung der zweiten Hand und ihr Verhältnis zur ersten sowie die Charakteristika, die die zweite Hand im vorliegenden Band zeigt: Die zweite Hand, von Roland (288) als „Gradualmaler B“ bezeichnet, auch in den Handschriften Brixen, Priesterseminarbibl., Cod. F. 4 als „Hand 1“ und Cod. E. 21 für gesamte Deckfarbenausstattung nachweis bar; Gestaltung der Binnenfelder im zweiten Gradualband jedoch wesentlich elaborierter; bei seinem Rand schmuck zeigt dieser Illuminator sowohl eine üppig-flächendeckende Variante (s. insbes. Cod. E.21, Bl. 14r) als auch eine italianisierende Version mit Rankenstäben, die mit diversen Manschetten und häufig symme trisch angeordneten Motiven geschmückt sind (s. u.). Da diese Art des Marginaldekors beispielsweise auch den niederösterreichischen Randleistenstil (ca. letztes Viertel 14. Jh., s. Roland, einschlägige Einleitung in MeSch II 129–131) prägt, muss man nicht unbedingt von einer Beeinflussung des Illuminators vor Ort in Italien ausgehen, sondern kann auch eine indirekte Beeinflussung im süddeutsch-österreichischen Raum bzw. – und dies ist wahrscheinlicher – in Südtirol annehmen. Roland (ebda) geht davon aus, dass der „Gra dualmaler B“ der ersten Hand untergeordnet war. Am Anfang bzw. während der Arbeiten an der Deckfarben ausstattung des vorliegenden Gradualbandes dürfte dies jedoch umgekehrt gewesen sein. Möglicherweise haben beide Buchmaler zunächst eine „Probearbeit“ abgeliefert: die erste Hand die Einleitungsseite und die zweite Hand Bl. 27v, woraufhin man – auch wenn sich die erste Hand durch akkuratere Arbeitsweise, reinere Farbgebung und delikatere Figurengestaltung auszeichnet – die lebendigere zweite Hand, die bei der Ausführung der Binnenfelder über die Hintergrundfolien der ersten Hand hinausgeht und versucht, mit ihren Architekturen und Landschaften räumliche Tiefe zu erzeugen, mit der Ausstattung und dem damit ver bundenen, relativ komplexen Bildprogramm des 1442 fertiggestellten Bandes betraute. Während die erste Hand auch Ornamentalinitialen gestaltete und die Akanthusranken überwiegend mit Tierdrolerien belebte, führte die zweite Hand ausschließlich figürliche Initialen aus, die in der Buchschmuck-Hierarchie über den ornamentalen Initialen stehen, und fügte darüber hinaus bei den Initialen der Feste Weihnachten, Epiphanie, Ostern und Kirchenweihe (Bl. 27v, 36v, 156v, 261r) in die Buchstabenkörper und in den Marginaldekor eine beachtliche Anzahl alt‑ und neutestamentarischer Verweise und Zusatzszenen ein (abweichend: Medaillon mit der Schweineschlachtung, Bl. 156v, das an Monatsbilder erinnert, dort allerdings zumeist dem Dezem ber zugeordnet). Sind Weihnachts‑, Oster‑ und Kirchweih-Initiale (jeweils drei Notenzeilen hoch) durch Binnenfelder herausgestellt, die an Bildteppiche mit übereinander geschichteten bzw. ineinandergreifenden Szenen erinnern, so werden bei der Epiphanie-Initiale (Bl. 36v), die lediglich zwei Notenzeilen hoch ist und keine figurative Buchstabenkörperfüllung aufweist, die typologischen Verweise in das Binnenfeld integriert. Zudem findet sich auf dem Blattgold-Außengrund dieser Initiale Fleuronnée-Dekor, der von der ersten Hand stammt (s. unter „Fleuronnée“); demnach assistierte in dieser Ausstattungsphase die erste Hand der zweiten. Im Folgenden die Charakteristika der zweiten Hand: 362 Graduale Neocellense I Schlichte Initialrahmen, Bl. 33r lediglich violette Linie. Außengründe bzw. -zwickel mit Blattgold belegt (tw. abgeblättert) und mit weißlichem Malmittel verziert; dabei wohl nicht nur der Fleuronnée-Dekor Bl. 36v von der ersten Deckfarbenhand, sondern auch die zarten Ranken wie bei der Initiale Bl. 261r. Stämmige Buchstabenkörper mit feinen Konturstegen (nur Bl. 27v Buchstabenkörper von breiterem, mit gelben Zier kreisen dekoriertem Rahmen eingefasst) sowie fallweise mit kleinen Blattausläufern und Manschetten in Form von (andersfarbigen) stilisierten Blüten (z. B. Bl. 261r); Bl. 27v steht der grüne Buchstabenschaft, an dem inwendig eine violette Zierleiste mit Golddekor anliegt, auf einem kleinen, stilisierten Drachen in Orange rot, der in den Letternschaft beißt. Bl. 27v, 33r und 156v Buchstabenkörper mit diversen Ton in Ton gemalten figurativen Szenen (Beschreibung s. u. in Auflistung), ansonsten mit tendenziell symmetrisch angelegtem Blattwerk gefüllt, dessen Einzelbl. sich stark eindrehen können; Bl. 36v die vegetabile Füllung des violetten Buchstabenkörpers unfertig, Vorzeichnung (?) sichtbar; bei dieser E-Initiale Querstrich mit andersfarbigen und ‑formigen Blättern geschmückt; Bl. 227r mittig im linken Buchstabenschaft kleines Köpfchen, dem Füllranken entwachsen. In Binnenfeldern charakteristische blau oder violett „marmorierte“ Hintergründe (tw. mit Farbverläufen), die Himmel oder Ausblicke bezeichnen (besonders aufwändig Bl. 36v und 200r); Bl. 227r als Hintergrund blaugrundiges Kachel‑ bzw. Teppichmuster. Die eher dünnen, (fast) zweiseitig bis (annähernd) vierseitig angelegten Äste der Marginalranken – sowohl lichte, in den Blattecken rechtwinklig geführte Rankenstäbe mit symmetrischen Blattpaaren und Fantasie blüten (z. B. Bl. 227v) als auch dichtere Voluten – gehen zumeist von den Serifen aus, Bl. 36v von der Man schettenblüte; diverse Blatttypen, häufig Paare von kleinen, spitzen Blättern sowie mehrbahnige Blätter, bei denen sich eine Bahn stark einrollt (vgl. Buchstabenkörperfüllungen); im Umfeld von Drolerien üppigere, tw. an drapierte Stoffbahnen erinnernde Blätter; Verzierung der Rankenäste durch Manschetten in diversen Ausformungen, Goldpunkte und -tropfen (in Astgabelungen). Auf allen Deckfarbenseiten Drolerien, zumeist anthropomorph (nur Bl. 227r ausschließlich Tierdrolerien wie bei der ersten Hand, s. Bl. 1r) und häufig in Rankenvoluten eingefügt; Marginaldekor Bl. 1r und 261r in der rechten unteren und Bl. 156v in der linken unteren Ecke abgegriffen und daher schlecht erkennbar. Eher ungelenke Figuren mit diversen anatomischen Unsicherheiten, beispielsweise Knieposition der Gottes mutter (Bl. 27v) oder des Königs, der dem Jesuskind seine Gabe überreicht (Bl. 36v), des Weiteren gum miartige, nackte Gliedmaßen (z. B. Bl. 33r rechtes Bein des Rankenkletterers sowie Arme und Beine des horizontal am Rankenast hängenden Putto, Bl. 200r, Bas-de-page) oder auch der flächige Brustkorb des Auferstandenen Bl. 156v; besonders unsicher die Darstellung von Rückenfiguren, z. B. die beiden Engel mit Notenblättern im Vertikalschaft der Initiale Bl. 156v; bei Frauenfiguren kleine, kugelige Brüste (z. B. Maria Bl. 27v oder Königstochter Bl. 27v, unterer Seitenrand). Gesichter von Älteren mit akzentuierenden Falten linien und charakteristischer Kerbe über der Nase (z. B. Bl. 33r: Priester und Rankenkletterer); Antlitz der Gottesmutter Bl. 27v folgt zwar dem üblichen Gesichtsschema (konturierte Nase, relativ kleiner Mund, run des Kinn), wirkt aber durch die zurückgenommene Augenlinierung und die hohe, gewölbte Stirn besonders lieblich; geöffnete Münder müssen nicht, wie sonst oft der Fall, für Verkommenheit stehen, sondern drücken vielmehr mündliche Rede aus (z. B. Prophetenreihe, Bl. 156v, oberer Foliorand) oder Gesang (z. B. ebda, linke untere Ecke, hier Zahnandeutungen, vgl. Propheten, abgerieben); Haare in der Regel einfach gesträhnt, mit relativ dicken Flechten. Unaufgeregte Draperien mit Röhren‑ und Knickfalten, lediglich Umhang des Auferstandenen Bl. 156v etwas aufwändiger gefältelt; sich abzeichnende Gliedmaßen mit Weißhöhungen herausgearbeitet, z. B. Beine und Knie Mariens, Bl. 27v. Bei Stoffen und Preziosen wenig Detailfreude (s. z. B. Bas-de-page Bl. 27v die lediglich mit schwarzen Kontur‑ und Binnenlinien gestaltete Goldrüstung des Drachenkämpfers oder den Bischofsstab, Bl. 261r), einfache Holzsichtigkeit (z. B. Holzmaserung des Beschneidungstisches Bl. 33r); lediglich die für diesen Buchmaler typischen „marmorierten“ Flächen, die Himmel bzw. Weite, aber auch kostbaren Stein bezeichnen (z. B. grüne Säulen Bl. 36v), von einiger Delika tesse; recht fein sind auch Notenblätter dargestellt, s. v. a. im Buchstabenkörper Bl. 156v. Vorliebe für Architekturen, die sehr detailreich (gerne mit Rundbogen) gestaltet sein können; dabei perspek tivische Unsicherheiten, so setzt bei der Kirche Bl. 261r die Giebelfront des Seitenschiffs direkt am Haupt portal an, oder bei dem Mauervorbau Bl. 33r (am unteren Binnenfeldrand) scheint das linke Mauerstück zu kippen; charakteristische Baumdarstellungen mit Kronen aus konzentrisch angeordneten Blättern, in deren Mittelpunkt jeweils ein roter Apfel (?) sitzt; Felsen (und Erde?) durch dunkelgraue Schollen wiedergegeben; der Fluss Bl. 261r kompakt wie ein Garnstrang. Insgesamt gedecktere (etwas nachgedunkelte?) Farbigkeit, speziell bei Marginalranken, mit gebrochenen Blau‑, Grün‑ und Violetttönen; allerdings häufig auch Orange verwendet, das recht leuchtend sein kann; Vorliebe für Violett (Rosa), u. a. für (einfachen) Baustein; auffällig die violetten Himmel Bl. 156v, 200r und 261v; Bl. 156v die ockerfarbene Initiale in Pinselgold ausgeführt; wenn Blattgold (v. a. Außengründe der Initialen) abgerieben, rosafarbener Bolus gut erkennbar. Bl. 1r König David, der seine Seele zu Gott erhebt: David kniet (?) betend im unteren Binnenfeld; Haltung der zum Gebet gefalteten Hände unklar wiedergegeben; aus seiner Brust entweicht relativ großes Seelen figürchen (ebenfalls mit gefalteten Händen), zu dem David aufblickt; im oberen Binnenfeld segnender Graduale Neocellense I 363 Gottvater (als Dreiviertelfigur) mit Weltkugel in der Linken. Drolerien konzentrieren sich auf Bas-de-page; erkennbar sind (von links nach rechts): dunkelblau-gelber (?) Vogel, „lächelnder“ Hase, Storch mit Schlange (?) im Schnabel, grüner Papagei, mittig Neustifter Wappen: goldenes, bedorntes Tau-Kreuz auf rotem Schild; darüber Blattranke mit Profilgesicht, rechts daneben kugelige Maus (?), des Weiteren stehender Bär mit Dudelsack und rotes Eichhörnchen. Bl. 27v Im Binnenfeld Geburt Christi und Verkündigung an die Hirten; über dezent bewachsenen Erdschol len, die nach unten bzw. vorne von einem durch wenige Bäume bezeichneten Garten und einen Flechthag ab gegrenzt werden, über den drei Hasen springen, erhebt sich rosafarbene Stadt mit bewehrter Mauer, Türmen, Treppengiebeln, hohem rundbogigen Tor und größerer Kirche (mit aufwändigem Portal), deren Chorturm (?) und Glockentürmchen Kuppelhauben tragen; diese repräsentieren in der Regel orientalisch-fremdländische Architekturen und sind an dieser, in ihrem restlichen Aufbau durchweg abendländischen Kirche wohl als Reminiszenz an Bethlehem zu verstehen; darüber Maria (groß angelegt und die Komposition beherrschend), die das in einer mit Heu gefüllten Krippe liegende Jesuskind kniend anbetet; Kind hält roten Apfel in der Linken und hat die Rechte zu seiner Mutter erhoben; hinter der Krippe roter Engel (mit Spruchband In dulcis [!] iubilo, singet und seyt fro, m[eines?] h[erzens] w[onne] l[eit] i[n] p[raesepio]), daneben Ochse und Esel, die von dem Heu fressen; hinter der Gottesmutter Stall (mit verwinkeltem Strohdach), in dem hinter einem violetten Vorhang Josef schläft; darüber baumbestandene Felslandschaft mit Schafen, in der ein weiterer roter Engel (als Büste auf stilisiertem Wolkenband, vom dem kompakte Strahlen ausgehen, mit Spruchband Ecce ewangeliso vobis gaudium m[agnum], Lc 2,10) drei Hirten (mit zwei Hunden, Hirtenstab, Fässchen und Spruchband Transeamus usque Bethle[hem], Lc 2,15) die Geburt Christi verkündet; die vertikale Mittel achse des Bildfeldes betonend reckt sich ein Widder (?), der seine Vorderhufe an einem Baum abstützt, zu dessen Blätterkrone hinauf; im Himmel (auf einem Wolkenband) fünf rote Engel, die sich um ein Notenblatt scharen (darauf Gloria in excelsis Deo et in terra erkennbar). – Im Schaft des Buchstabenkörpers drei Figuren: oben und unten eine mit erhobenen Händen betende Frau, jeweils von hinten gegeben, in der Mitte ein von vorne zu sehender Engel (Flügel nicht eindeutig erkennbar, es könnte sich auch um Blattpaar handeln, vgl. die anderen Blätter der Schaftfüllung) mit einem Spruchband in der Linken und erhobener Rechter mit ausgestrecktem Zeigefinger; im P-Bogen frontal thronender König David (?) mit Zepter in der Linken und Spruchband in der Rechten, zu seinen Füßen ruhender Löwe, darunter Teufel mit Eselsohren. – Marginal schmuck: auf Initialhöhe zwei Männchen (blau und violett), die übereinander in der Ranke sitzen und auf einander deuten, wobei sich die obere Figur mit der Linken an draperieartigem Blattausläufer festhält; gänz lich in die Ranke integriert, die hier eine starke, mit kleinen Kreisen verzierte Mittelrippe aufweist, ein kopfloser grüner Drache mit violettem Flügel; direkt darunter ein kleinerer orangefarbener Drache; auf der Höhe des P-Schafts in Rankenvolute hockend der greise Prophet Jesaja mit violetter Kappe und Spruchband (Ecce virgo concipiet et par[iet] fi[lium], Is 7,14); auf dem unteren Seitenrand Drachenkampf: Ritter in Goldrüstung auf Schimmel (hl. Georg?) stößt einem grünen Drachen mit orangefarbenen Fledermausflügeln seine zersplitternde Lanze ins Maul, dem nicht nur Feuerzungen, sondern auch die Bas-de-page-Ranken entwachsen; in Rankenmedaillon zwischen den Kontrahenten eine kniend (?) betende Frau mit Krone (Königstochter, die von Georg befreit wurde?). Bl. 33r Im Binnenfeld drastisch dargestellte Beschneidung: In farbfreudigem Architekturrahmen (mit Säu len, unterschiedlich ausgeformten Bögen und kassettierten Decken), der Himmelsausblicke freigibt, be schneidet jüdischer Priester (Mochel) mit einer Schere das auf einem breiten Holztisch liegende, relativ große Jesuskind, das von vier weiteren Priestern (einer davon mit Krücke) und einem Mann mit Judenhut festgehalten wird; zu dieser Szenerie führt eine Treppe hinauf, welche zu einer Art Stadtmauer gehört; auf einem Vorsprung dieser Mauer zwei Frauen, die nach oben zum Beschneidungstisch blicken. – Im Buch stabenkörper links Geburt und Waschung Christi: Ein Engel hebt den Vorhang über dem Bett, auf dem Maria liegt und auf Josef und ihr Kind blickt; Josef mit Stock und Fässchen (?); das Kind in einem von Engeln umgebenen Weidenbottich sitzend; darunter weiterer Engel und eine weibliche Figur (Engel?) von hinten; rechts im Buchstabenkörper zwei weitere Engel, jeweils mit Spruchband; in der Serife vier Engelsfiguren bzw. -köpfe. – Marginalschmuck: auf Rankenspitze Männchen mit Krone, der zwei Blattzipfel entwachsen; diese Figur hält sich, auf den Schultern eines weiteren Männchens hockend, mit seiner Linken an der nächst liegenden Fleuronnée-Initiale fest und deutet mit der Rechten auf den geöffneten Mund seines Trägers, dessen Beine bzw. Füße in den Rankenstab übergehen; auf der Höhe der Deckfarbeninitiale weiterer Ranken kletterer mit ebenfalls geöffnetem Mund und roter Haube, nach dessen Stiefel von unten her ein nacktes Männchen mit Lendentuch und grünem Umhang greift; links auf Bas-de-page in einem Blattpaar blaue Büste, die mit einem Stock eine orangefarbene Figur attackiert, welche mit einem Ausfallschritt und sich an einem Ast festhaltend die Verbindung zw. zwei Rankenabschnitten herstellt. Bl. 36v Im Binnenfeld Epiphanie mit typologischem Charakter: Im oberen Binnenfeld links, in Fels landschaft mit drei burgartigen Städtchen, die Heiligen Drei Könige, die auf das Kind im Strahlenkranz (auf Felsspitze, rotes Äpfelchen in der Linken) und damit auf die Worte des Bileam (Nm 24,17) weisen, die das Kind als Spruchband in der Rechten hält Orietur stella ex [Iacob …] (Hermann liest zudem: in Iudeam et 364 Graduale Neocellense I invenietis); tw. verdeckt von der Felsnadel, auf der das Kind sitzt, haben sich die Könige auf ihren Pferden nach rechts gewandt, in Richtung eines kleinen rundbogigen Tores. Im unteren Binnenfeld auf reich verziertem Holzthron Maria mit dem Kind, das die Hände nach den Gaben der Heiligen Drei Könige ausstreckt; links neben dem Thron schläft Josef auf seinen Stock gestützt, hinter ihm das rundbogige, hier größer angelegte Tor, das die obere mit der unteren Bildebene und somit die alttestamentarische Prophe zeiung des Jakobssterns mit dem neutestamentarischen Stern der Heiligen Drei Könige verbindet. Diese Verknüpfung wird noch durch den mittleren König unterstrichen, der auf das Tor hinter Josef bzw. nach oben weist. – Marginalschmuck: Büste eines greisen Königs (David), der auf die Deckfarbeninitiale deutet und Spruchband mit dem Psalm Reges Tarsis (Ps 71,10) hält, welcher ebenfalls auf die Anbetung der Heiligen Drei Könige verweist. Auf der Höhe der Fleuronnée-Initiale hockende Figur in Blaugrau, die in Richtung Initiale weist; grüner Drache mit verschlungenem Hals, der dem oberen Profilkopf des Fleuronnée-Dekors in die Nase beißt; gräulicher Bär mit Dudelsack (?); grauer Kopf, der aus eingerolltem Blatt wie aus einem Mantelkragen herausblickt; auf Bas-de-page vier figurierte Voluten (von links nach rechts): Prophet Ezechiel (Halbfigur) mit Spruchband Et effundam super eos aquam mundam (Ez 36,25); Taufe Christi; Prophet Baruch (knappe Dreiviertelfigur) mit Spruchband Post hec in terris visus est (Bar 3,38); Hochzeit zu Kana: Maria mit Spruchband Vinum non habent (Io 2,3) und Jesus, der mit der Rechten auf sechs blaue Weinkrüge deutet und mit der Linken Segensgestus ausführt. [Bl. 129r Einzug in Jerusalem, Ranke mit Brustbildern] Bl. 156v Im Binnenfeld Auferstehung Christi: In einem von einem Flechtzaun umschlossenen Garten steht der Auferstandene, dessen Wundmale deutlich zu sehen sind, frontal auf dem geschlossenen Sarkophag (mit Halteringen auf dem Deckel und versiegelt?), die Rechte segnend erhoben, in der Linken die Siegesfahne; zu seinen Füßen die schlafenden Soldaten; das felsige Terrain von einigen Bäumen bestanden (an einem nagt, rechts unten neben Christus, ein kleiner Hase), links wölben sich die grauen Felsen grottenartig über den Sarkophag; zwei kleinere Stadtveduten mit Türmchen. – Im Buchstabenkörper links drei Engel, die drei Notenblätter mit österlichen Gesängen präsentieren (von oben nach unten): Salve festa dies toto venerabi lis evo, Regina celi letare und Alle[luia] Dei filius ab hoste superbissimo resurgens; rechts oben Noli me tangere: in einem Garten kniet Maria Magdalena (mit Salbgefäß) vor Christus und weist mit der Rechten auf die Wundmale seiner Füße; rechts unten Drei Frauen am Grabe: in einem Garten zeigt ein Engel den drei Marien den offenen und leeren Sarkophag. – Marginalschmuck, jeweils in Rankenvoluten: auf dem oberen Seitenrand siebenmal der Prophet Hosea, jeweils mit Spruchband (Verse Os 6,1–3 lassen sich auf Auferste hung beziehen): Venite et revertamur ad dominum (Os 6,1), Quia ipse cepit et sanabit nos (Os 6,2), Percutiet et curabit nos (Os 6,2), Vivificabit nos post duos di[es] (Os 6,3), In die tertia suscitabit nos (Os 6, 3), Et vivemus in conspectu eius (Os 6,3), Sciemus sequemurque ut cognoscamus dominum (Os 6,3); auf dem linken Seitenrand (oben) vier stehende Engel, jeweils mit Spruchband (Verse eines deutschen Ostergesanges, von oben nach unten): Maria du zarte, Du bis ayn rosengarten, Den got selwer geczyrt hat, Wen her von dyr geporen wart; darunter fünfmal der Prophet Jesaja (sitzend, mit langem grauem Haar und Bart), jeweils mit Spruchband (von oben nach unten): Quis est iste qui venit de Edom (Is 63, 1), Iste formosus in stola gradiens (Is 63, 1), Torcular calcavi solus de gen[tibus] (Is 63, 3), Dies enim ulcionis in corde meo (Is 63, 4), Annus retribucionis mee venit (Is 63, 4); auf Bas-de-page (von links nach rechts; abgerieben): zwei sitzende, singende Männer mit Notenblatt (Vespere autem schabati [que] lu[cescit], Mt 28,1), zwei Männer, die Schwein schlachten, je ein Prophet mit Spruchband (Beschriftung des ersten nicht mehr lesbar, auf dem zweiten nur noch gentes et zu erkennen), Lamm mit Siegesfahne (dieses Motiv mit erhaltenem Kolorit in Cod. E.21, Bl. 29v, vgl. die identische Haltung der ungelenk verschränkten Vorderbeine). Bl. 200r Im Binnenfeld Himmelfahrt Christi: Hinter einem Flechthag ragt der an seinem Fuß und auf dem Gipfel baumbestandene Auffahrtsfelsen empor, um den sich Maria und Apostel betend geschart haben; vio letter Himmelsgrund, in dem Engelspaar auf Wolkenbändern schwebt, jeweils mit Spruchband (Viri Galilei quid admiramini / [Asp]icie[nte]s in celum, Apc 1,11); darüber stilisiertes Wolkenband mit Ton in Ton ge malten orangefarbenen Engeln, die den Auffahrenden umgeben, von dem noch Füße (mit Wundmalen) und Gewandsaum zu sehen sind; auf dem Felsen sind Fußspuren erkennbar. – Drolerien: auf dem rechten Seiten rand heraldisch aufsteigendes gelbes Einhorn (vgl. sehr ähnliche Ausführung dieses Fabeltiers in Cod. E.21, Bl. 14r, links auf Bas-de-page), mittig auf Bas-de-page Drachenpaar (orange und grün) mit verschlungenen Köpfen, flankiert von Putti, die in Ranken bzw. auf Rankenstäben sitzen oder an ihnen hängen und sich gegenseitig mit Stöcken attackieren (Turnierpersiflage). [Bl. 218r In festo ss. trinitatis (zweimal)] Bl. 227v Binnenfeld von Vorhang horizontal in zwei Bereiche aufgeteilt: Vorne kniet auf Rasenstück ein Augustiner Chorherrenpropst im hellen Chorherren-Mantel mit grauem Kapuzenkragen (mit Schweifen be setztes Feh-Almutium), mit violetter Kappe und Spruchband (Domine in tua misericordia speravi exultavit cor meum) und betet Schmerzensmann an; dieser als Dreiviertelfigur in Sarkophag (zw. Vorhang und teppichartigem Hintergrund), Hände vor der Brust überkreuzt, mit Dornenkrone, Rutenbündel und Geißel. – Drolerien: auf dem linken Seitenrand Fabelwesen mit Drachenkörper und Vogelkopf, dessen Hals sich um Graduale Neocellense I 365 den Rankenstab windet, in den er zusätzlich mit seinem langen Schnabel beißt; auf Bas-de-page zwei graue Häschen. Bl. 261r Im Binnenfeld Kirchweihe: Im unteren Feldbereich hölzerner Steg, der flankiert von geradstäm migen Bäumen über einen Fluss führt; dahinter Mauer, über der sich Kirche mit Seitenschiff (goldene Krab ben auf Giebeln) und Turm erhebt, von dessen Satteldach die prächtige Kirchweihfahne (Kreuzfahne) weht; vor der Kirche (neben einem Baum) Bischof im Ornat mit großem, goldenem Krummstab; hinter der Mauer bzw. über dem Kirchenareal dicht stehende Bäume, vor denen (auf stilisiertem Wolkenband) segnender Christus mit Spruchband (Ista tenella florens ut rosa Nova Cella) schwebt; hinter ihm schmaler violetter Himmelsstreifen. – Marginalschmuck: auf Bas-de-page, jeweils in zarte Rankenmedaillons eingeschrieben, Strahlenkranzmadonna (Mutter und Kind mit roten Heiligenscheinen, Kind hält Äpfelchen? in der Rechten), flankiert von zwei adorierenden Klerikergruppen; links fünf Augustiner Chorherren, zuvorderst ein Chor herrenpropst mit Schweifen am Feh-Almutium, roter Kappe und Spruchband (Nos cum prole pia benedic[at] vi[rgo] M[aria]), etwas abgesetzt von der Gruppe Mönch in dunkler Kutte (Franziskaner?) mit Spruchband (Suppliciter te petimus); rechts von Strahlenkranzmadonna weitere fünf Chorherren, der nächste zur Gottes mutter mit Spruchband (Ora pro nobis mater mi[sericordi]e); auf dem rechten Seitenrand (von unten nach oben): Christus und Zachäus, auf zwei Medaillons aufgeteilt; unten Christus mit Gefolgschaft (drei Jünger, zwei Juden, zwei Frauen); Spruchband Jesu (Zachee festinans descende, Lc 19,5) schlägt Brücke zum obe ren Medaillon, in dem der Zöllner Zachäus in den Bäumen steht; in Rankenvolute darüber schläft Jakob mit dem Kopf auf einer Bodenandeutung (ansonsten keine Auflagefläche angegeben, sodass Jakob zu schweben scheint), über ihm gelbe Leiter (am Fuß flankiert von zwei Spruchbändern: Vidit Jacob scalam und Et sum mitas eius tangebat celum, Gn 28,12), auf der fünf Engel (die beiden unteren von vorne, die drei oberen von hinten zu sehen) hinauf zu Christus klettern, der als Büste auf stilisiertem Wolkenband die Rechte segnend erhebt und in der Linken ein Spruchband (Benedicens benedicam tibi terram in qua dormis dabo et …, Gn 28,13) hält. Zur Interpretation dieser Seite s. zweiter Gradualband unter „Bildnisse“. Bildnisse. Die angeführten Darstellungen sind nicht als Porträts im Sinne getreuer Abbilder mit individuel len Zügen zu verstehen, sondern als bildliche Repräsentanten der betreffenden Persönlichkeiten. Der an betende Augustiner Chorherr Bl. 227v, den der Fehschweifbesatz seines Almutiums und die violette Kappe als Propst auszeichnen, dürfte – wie schon Hermann (223) annahm – den Auftraggeber, Propst Nikolaus Scheiber darstellen; ebenso der Chorherr, der auf dem Bas-de-page Bl. 261r die adorierende Gruppe zur linken Hand der Strahlenkranzmadonna anführt (s. Giner 10; vgl. zudem Darstellung im zweiten Band Bl. 210v). Wen der Mönch (Franziskaner?) darstellen soll, der sich Bl. 261r (Bas-de-page) zu den anbetenden Chorherren gesellt, ist bislang ungeklärt; es dürfte sich jedoch nicht, wie von Hermann (224) angenommen, um den Schreiber Friedrich Zollner handeln, da dieser Augustiner Chorherr war. S. R. E: Renaissance-Einband: dunkelbraun gefärbtes Leder über Holz mit Blinddruck. Neustift, 2. H. 16. Jh. Einband restauriert, Leder tw. ergänzt (bes. deutlich sichtbar am VD und HD zum Rücken hin sowie am Rücken), Bindung erneuert. VD und HD ähnlich, Gliederung jeweils durch dreifache Streicheisenlinien. Am VD fünf Rahmen (von außen nach innen): Rahmen 1 gefüllt mit Rolle einseitwändig verschränkte Bögen mit Lilienbekrönung, darüber Leiste aus fünfblättrigen Blumen, seitlich je eine Rolle, oben und unten zwei Rollen spiegelsymmetrisch angeordnet (Nr. 1); Rahmen 2 gefüllt mit Rolle Profilköpfe in Kreis im Wechsel mit Schriftband und begleitendem floralem Dekor (Nr. 2 = Neuhauser, Rolleneinbände, R 48); Rahmen 3 gefüllt mit Rolle Wappen von Neustift (Tau-Kreuz), Säben und Patriarchenkreuz einander ab wechselnd, verbunden mit Rankendekor in gegenständigen Dreipassformen (Nr. 3 = Neuhauser, Rollen einbände, R 58); Rahmen 4 gefüllt mit Rolle Profilköpfe in Handspiegeln mit Quastenbehang (Nr. 4 = Neu hauser, Rolleneinbände, R 22); Rahmen 5 gefüllt mit eng aneinander gereihten Einzelstempeln Schriftband „Maria“, dazwischen Fischblasenmotiv und in den Zwickeln Trifolien (Nr. 5). Mittelfeld in vier Einzelfelder unterteilt, die jeweils wieder zwei von je dreifachen Streicheisenlinien begrenzte Rahmen aufweisen (von außen nach innen): Rahmen 6 gefüllt mit Einzelstempel Maske frei (Nr. 6 = Neuhauser, Rolleneinbände, S 41) im Wechsel mit Lilie frei (Nr. 7); Rahmen 7 gefüllt mit Rolle (Nr. 3, s. Rahmen 3). Ecken des äußeren und inneren Rahmens jeweils mit Blindlinien verbunden, sodass in der Mitte des VD ein Andreaskreuz ge bildet ist. Mittig Einzelstempel Stempelblüte mit Rautenranke (Nr. 8 = Neuhauser, Rolleneinbände, S 39) mit eingeschlossener Lilie frei (Nr. 7). Spuren eines großen Mittelbeschlags mit sechs (?) Nagellöchern, Spuren von Eckbeschlägen mit je drei (?) Nagellöchern. Am HD fünf Rahmen (von außen nach innen): Rahmen 1 gefüllt mit Rolle ineinander greifende, aus drei Linien aufgebaute Bögen mit Lilienbekrönung und zwei eingeschlossenen vierblättrigen Blüten (Nr. 9); Rahmen 2 gefüllt mit Rolle (Nr. 1); Rahmen 3 gefüllt mit Rolle (Nr. 3); Rahmen 4 gefüllt mit Rolle (Nr. 2); Rahmen 5 gefüllt mit Rolle (Nr. 4). Mittelfeld gerahmt, Längsfelder frei, unten und oben jeweils drei Einzel stempel Lilie in Raute (Nr. 10), mittig gerahmte Raute eingeschrieben: Rahmen gefüllt mit Rolle (Nr. 4); im 366 Graduale Neocellense I Rautenmittelfeld Brokatmotiv (mittig zum Vierblatt angeordnet, oben und unten jeweils ein Brokatmotiv geschlossen) (Nr. 11), oben mit eingeschlossenem Schriftband (?), unten mit eingeschlossenem Einzel stempel florales Dekor, rautenförmig (Nr. 12). In der unteren Rautenhälfte zudem zwei seitlich angeordnete Einzelstempel florales Dekor mit geschlossener Knospe in Rautenform (Nr. 13). In den Außenfeldern der Raute oben und unten jeweils zwei am Rahmen angesetzte Brokatmotive offen (Nr. 14) sowie in der unteren Hälfte Einzelstempel (Nr. 13, Nr. 5 und Nr. 10), in der oberen Hälfte Einzelstempel (Nr. 5 und Nr. 10). Am HD vier Eckbeschläge in Form von steigenden gekrönten Löwen mit eckigen Buckeln. Holz am oberen, unteren und seitlichen Rand eingekerbt, durch jeweils sechsfache Streicheisenlinien betont. Zwei Langriemen-Leder-Metallschließen mit Tierkopfabschluss (wohl nicht original), Leder erneuert, am VD Haltedorn auf runden Platten. Am VD und HD jeweils zwei Stehkanten. Kanten gerade. Rücken: sieben Doppelbünde. Kapitale mit naturfarbenem und rotem Zwirn umstochen. Spiegel sowie Vor- und Nachsatzbl. Perg. 20. Jh., leer bis auf Etikett der Restaurierwerkstätte (s. bei G) am Spiegel des HD. G: Hs. im Auftrag des Propstes Nikolaus Scheiber (Propst 1439–1449) in Neustift angefertigt. Die zahlreichen Streichungen, Ergänzungen und Hinweise belegen eine Verwendung der Hs. bis ins 19. Jh. Hs. bei vorüber gehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. Zahlreiche Bl. (s. auch bei B, A und Inhalt) verloren, tw. erst zw. 1905 und 1965 (vgl. Beschreibung bei Hermann bzw. Laußermayer). Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro, S. Maria di Rosano [Firenze]), kein Restaurierbericht erhalten. L: Hermann 221–224, Nr. 222. – B. Rutz, Illuminierte Choralhandschriften zu Neustift. Kirchenmusikalische Jahrbuch 22 (1909) 88–95, hier 90–94. – Laußermayer 246–248, 257–267. – H. Lehmann-Haupt, Gutenberg and the Master of the Playing Cards. New Haven-London 1966, 35–36, 43. – G. Schmidt, Gotik in Öster reich. Ausstellung 19. Mai bis 15. Oktober 1967, Minoritenkirche Krems-Stein. Krems an der Donau 31967, 136 (auch in: M. Roland [Hrsg.], Gerhard Schmidt, Malerei der Gotik. Fixpunkte und Ausblicke I: Malerei der Gotik in Mitteleuropa. Graz 2005, 46). – W. A. Schulze, Alte Dreikönigsbilder in Südtirol. Der Schlern 53 (1979) 5–17. – W. Salmen, Katalog der Bilder zur Musikgeschichte in Österreich 1 (Innsbrucker Beiträge zur Musikwissenschaft IV). Innsbruck 1980, Nr. 269. – Peintner, Buchmalerei 14, 21, 43, 45, 49–57, 63– 64. – M. Peintner, Kloster Neustift. Augustiner Chorherren in Südtirol. Bozen 1985, 19. – N. Rasmo, Kunst schätze Südtirols, Rosenheim 1985, 132–133. – M. H. Peintner, Psallite Deo! Zur Musikgeschichte des Stiftes Neustift, in: 850 Jahre Augustiner Chorherrenstift Neustift. 134–153, hier 138–139. – Ch. Giner, Leben im Kloster, in: 850 Jahre Chorherrenstift Neustift. Katalog 85–90, hier 89. – M. Peintner, Das Skriptorium von Neustift, in: 850 Jahre Neustift. Katalog 91–93, hier 93. – M. Peintner, Musikgeschichte des Stiftes, in: 850 Jahre Neustift. Katalog 94–100, hier 94. – Th. (H.) Innerhofer, Wappen und Siegel des Konvents und der Pröpste von Neustift, in: 850 Jahre Neustift. Katalog 199–214, hier 199. – P. Caneppele, Ista tenella florens ut rosa Nova Cella. Una raffigurazione ideale della chiesa abbaziale di Novacella? Archi vio per l’Alto Adige 88/89 (1994/95) 356–366. – Gozzi 546, Nr. 45. – Stefani 120–121. – O. Lässer, Bilder zur Musikgeschichte in Tirol. Von den Anfängen bis 1600, in: K. Drexel, M. Fink (Hrsg.), Musikgeschichte Tirols I: Von den Anfängen bis zur Frühen Neuzeit (Schlern-Schriften 315). Innsbruck 2001, 705–769, hier 730, Nr. 70. – Neuhauser, Musikgeschichtliche Quellen 151, 161, 164, 194. – Engels 310. – J. G. Alexander, J. H. Marrow, L. Freeman Sandler, The Splendor of the Word. Medieval and Renaissance Illuminated Ma nuscripts at The New York Public Library. London 2005, hier 181. – M. Studničková, Vážka jako obraz duše hledající spásu. Výzdoba Bratislavského misálu IV (AMB EL 13a) a skupina iluminovaných rukopisů bri xenského, vídeňského a jihoněmckého původu z poloviny 15. století / Libelle als Sinnbild der heilsuchenden Seele. Die Ausstattung des Pressburger Missale IV und eine Gruppe illuminierter Handschriften aus Brixen, Wien und Süddeutschland aus der Mitte des 15. Jahrhunderts, in: D. Buran (Hrsg.), Galéria. Ročenka Slovenskej národnej galérie / Jahrbuch der Slowakischen Nationalgalerie in Bratislava 2004/2005. Príspevky Medzinárodného kolokvia Gotika na Slovensku a jej stredoeurópsky kontext / Beiträge des Internationalen Kolloquiums Gotik in der Slowakei und ihr mitteleuropäischer Kontext. Bratislava 2006, 149–159, insbes. 158. – Roland, Gotik 274, 288–289 (Kat. 185). – Beier I 15–21, insbes. 16. – Baroffio 325, Nr. 16756. – G. Gabrielli, Tropes in cantus planus sources from South Tyrol, in: J. Border u. a. (Hrsg.), Cantus Planus. Ven ice meeting 2014. Venezia (in Druck). – Rischpler. Graduale Neocellense. (Ir–v) leer. (IIr) Frontispiz (s. bei A). (IIv) leer. Graduale Neocellense I 1 367 (1r–260v) Temporale (pars hiemalis und pars aestivalis). (1r) Dnca 1. adventus. – (3r) Dnca 2. adv. – (5r) Dnca 3. adv. – (7r) Fer. 4. quattuor temporum. – (9v) Fer. sexta. – (11r) Sabbato quattuor temporum. – (13v) Canticum trium puerorum (Dn 3,57– 88). – (16v) Dnca 4. in adv. – (18r) In vigilia nativitatis dni. – (19v) In galli cantu. Mit Sequenz (AH 53, Nr. 10). – (22v) In aurora. Mit Sequenz (AH 53, Nr. 16). – (26v) Ad tertiam missam. Mit Tropus (AH 49, Nr. 1) und Sequenz (AH 53, Nr. 15). – (31v) Dnca 1. post nativitatem dni. Mit Verweis auf Sequenz (AH 53, Nr. 10). – (33r) In circumcisione dni. Mit Verweis auf Sequenz (AH 53, Nr. 16). – (35v) In vigilia epiphaniae dni. Nur Verweis auf Introitus: Dum medium silen cium (rot:) per totum. – In epiphania dni. Rubrik: In epiphania domini tropus (von anderer Hand hinzugefügt) non canitur. Introitus mit Tropus (AH 49, Nr. 77, 1, 3a, 2b und Nr. 11) und Sequenz (AH 53, Nr. 29, unvollständig). – (Bl. 38v) Expl. mut.: … O Christe quantum patri exercitum. Iuvenis doc[tus (1 Bl. fehlt) (40r) Inc. mut. (Forts. der Sequenz): ausculta]te populi preceptori … – (40v) Dnca 1. post epiph. dni. Expl. mut. (Introitus): … angelorum psallentes in unum ecce cuius [ (2 Bl. fehlen, Textver lust) (43r) Inc. mut. (Communio): ] tuus dolentes querebamus te et quid est quod me querebatis … – Dnca 2. post epiph. dni. – (45v) Dnca 3. post epiph. dni. – (47v) Dnca in Septuages. – (50r) Dnca in Sexages. – (52v) Dnca in Quinquages. Expl. mut. (Introitus): … Esto mihi in Deum am[ (1 Bl. fehlt) (54r) Inc. mut. (Graduale): ]um filios Israhel et Ioseph … – (55v) In capite ieiunii: Ad processio nem. Mit Antiphonen Exaudi nos domine (CAO Nr. 2770), Immutemur habitu (CAO Nr. 3193), Iuxta vestibulum (CAO Nr. 3554). – (56v) Ad missam. – (59v) Fer. 5. post Quinquages. – (60v) Fer. 6. post Quinquages. – (62r) Verweis Sabbato ad missam Esto mihi (rot:) per totum. – (62r) Dnca 1. in Quadrages. – (66v) Fer. 2. – (68r) Fer. 3. – (69v) Fer. 4 quattuor temporum. – (72r) Fer. 5. – (74r) Fer. 6. – (75v) Sabb. – (77r) Dnca 2. in Quadrages. – (77v) Fer. 2. – (79r) Fer. 3. – (80r) Fer. 4. – (80v) Expl. mut. (Graduale): … Salvum fac populum tuum domine [ (6 Bl. fehlen) (87r) Sabbato post dncam 2. in Quadrages. Inc. mut. (Offertorium): ob]dormiam in morte ne quando dicat inimicus meus … – Dnca 3. in Quadrages. – (90v) Fer. 2. – (92v) Fer. 3. – (94r) Fer. 4. – (95v) Fer. 5. – (96v) Expl. mut. (Versus von Graduale): … Aperis tu manum tuam et imples [ (1 Bl. fehlt) (98r) Fer. 6. – (99v) Sabb. – (101r) Dnca 4. in Quadrages. – (104r) Fer. 2. – (105r) Fer. 3. – (107v) Fer. 4. – (110r) Fer. 5. – (112r) Fer. 6. – (114r) Sabb. – (116r) In passione dni. – (119v) Fer. 2. – (121v) Fer. 3. – (123r) Fer. 4. – (125v) Fer. 5. – (127v) Expl. mut. (Communio): … michi spem dedisti hec me consolata est in humilitate me a[ (2 Bl. fehlen: Es fehlen die Prozessionsgesänge des Palmsonntags, s. auch bei B und A.) (130r) Dnca in palmis. Inc. mut. (Introitus): ] facias auxilium tuum a me ad defensionem meam … – (136r) Fer. 2. Am Rand von späterer Hand (18. Jh.) ein Kyrie-Ruf ergänzt. – (137v) Fer. 3. – (140r) Fer. 4. – (143v) Expl. mut. (Offertorium): … Domine exaudi orationem meam et clamor meus ad [ (8 Bl. fehlen: Es fehlt beinahe das gesamte Triduum.) (152r) Sabbatum sanctum. Inc. mut. (Tractus): ] Hic Deus meus et honorabo eum Deus patris mei … – (155v) Ad vesperas. – (156r) Ad processionem: Mit Antiphon (CAO Nr. 3222). – (156v) Dnca resurrectionis dni. Mit Sequenz (AH 53, Nr. 36, unvollständig). – (158v) Expl. mut. (Str. 3): … Qui se ipsum exinanivit ut nos [ (4 Bl. fehlen) (163r) Dnca resurrectionis dni. Inc. mut. (Communio): epule]mur in azimis sinceritatis et ve ritatis. – (163r) Fer. 2. Mit Sequenz (AH 53, Nr. 46). – (167v) Fer. 3. Mit Sequenz (AH 53, Nr. 50). – (171r) Fer. 4. Mit Sequenz (AH 53, Nr. 52). – (174v) Fer. 5. Mit Sequenz (AH 53, Nr. 53). – (178v) Fer. 6. Mit Sequenz (AH 53, Nr. 54). – (182r) Sabbato. Mit Sequenz (AH 53, Nr. 56). – (185r) Dnca in albis = Dnca 1. post pascha. Mit Sequenz (AH 54, Nr. 7). – (187v) Fer. 2. – Fer. 3. – (187bisr) Fer. 4. – (187bisv) Kyrie-Ruf. Pascale minus. – (188r) Fer. 4. – Fer. 6. – (188v) De lancea dni. Mit Sequenz (AH 54, Nr. 140). – (191v) Dnca 2. post pascha. Mit Verweis 368 Graduale Neocellense I auf Sequenz (AH 54, Nr. 7). – (193r) Dnca 3. post pascha. Mit Rubrik von späterer Hand: Verweis auf drei Sequenzen (AH 54, Nr. 7; AH 54, Nr. 148; AH 50, Nr. 240). – (195r) Dnca 4. post p ascha. Mit Verweis auf Sequenz (AH 54, Nr. 148). – (195v) Dnca 5. post pascha. Mit Verweis auf Sequenz (AH 50, Nr. 240). – (196v) Rogacionum. – (198v) In vigilia ascensionis dni. – (199v) In die sancto. – (200v) Expl. mut.: … (Versus) Ascendit Deus in iubilacione et dominus in voce tube. Alleluia. (rot:) V. [ (6 Bl. fehlen: Es fehlen beinahe die gesamten Gesänge des Pfingstfestes sowie die entsprechenden Prozessionsgesänge „Rex sanctorum angelorum“, „Salve festa dies“.) (207r) Dnca pentecostes et de spiritu sancto. Inc. mut. (Alleluia. Versus): ] faciem terre. Alleluia. Mit zwei Sequenzen (AH 53, Nr. 70; AH 54, Nr. 153). – (212r) Fer. 2. Mit Verweis auf Sequenz (AH 53, Nr. 70). – (213r) Fer. 3. Mit Verweis auf Sequenz (AH 53, Nr. 70). – (214v) Fer. 4. Mit Verweis auf Sequenz (AH 54, Nr. 153). – (215v) Fer. 5. Mit Verweis auf Sequenz (AH 54, Nr. 153). – (217r) Sabbato quattuor temporum. – (217v) In festo ss. trinitatis. Expl. mut.: … Alle luia. (rot:) De sancta trinitate Introitus [ (1 Bl. fehlt) (219r) Inc. mut. (Alleluia. Versus): ]ia. Benedictus es domine Deus patrum nostrorum … Mit Sequenz (AH 53, Nr. 81, unvollständig). – (219v) Expl. mut. (Sequenz, Str. 7): … Proprietas in personis unitas est et in essencia. Maiestas par et po[testas (1 Bl. fehlt) (221r) Inc. mut. (Sequenz, Str. 17): aeter]nitas. Per te sumus redempti summa tu caritas … – (222r) In festo ss. corporis Christi. Mit Sequenz (AH 50, Nr. 385) und Offertorium mit Prosula (AH 43, Nr. 643). – (227r) Dnca 1. post trinitatis. – (229r) Dnca 2. – (230r) Dnca 3. – (231r) Dnca 4. – (232v) Dnca 5. – (234r) Dnca 6. – (235v) Dnca 7. – (237r) Dnca 8. – (238v) Dnca 9. – (240r) Dnca 10. – Dnca 11. – (241v) Dnca 12. – (243r) Dnca 13. – Dnca 14. – (246v) Dnca 15. – (247v) Dnca 16. – (248v) Dnca 17. Expl. mut. (Introitus): … secundum misericordiam tuam. Beat[i (1 Bl. fehlt, Foliierung durchgehend, s. bei B) (249r) Communio. – (249r) Fer. 4 quattuor temporum. – (250v) Fer. 6. – (251r) Sabbato. – (251v) Dnca 18. – (253r) Dnca 19. – (253v) Dnca 20. – (254r) Dnca 21. – (256v) Dnca 22. – (258v) Dominica ultima = Dnca 23. – (260v) In dedicatione ecclesiae. Mit Sequenz (AH 53, Nr. 247). – (264v) In dedicatione unius altaris. Expl. mut. (Introitus): … munera tua accepta erunt super altare meum. Miseri[ (2 Bl. fehlen) (267r) Inc. mut. (Ende der Sequenz AH 53, Nr. 112): gau]diorum vestrorum. (267r) Pro defunctis. – (268v) Expl. mut. (Offertorium): … libera animas omnium fidelium defunctorum de manu inferni et de profundo lacu libera eas de ore leo[nis (1 Bl. fehlt) (270r) Inc. mut. (Offertorium): ] eas in sinum Abrahe in lucem sempiternam. (rot:) Communione. (schwarz:) Absolve domine animas … – (270v) Pro pontificibus et praelatis defunctis. – (271r) später hinzugefügt: Dom. 2. Quad. 2 2.1 (271v–310v) Kyriale. (271v–290v) Kyrie. Gloria. (271v) Ohne Tit. Nur Kyrie. – Aliud, am Blattrand von späterer Hand ergänzt: De Apostolis 1. – (273v) Ohne Tit. Nur Kyrie. – Aliud, am Blattrand von späterer Hand ergänzt: De Confessoribus 2. Nur Kyrie. – (274r) Aliud Kyrie, am Blattrand von späterer Hand ergänzt: Angelicum 3. – Aliud, am Blattrand von späterer Hand ergänzt: De Martyribus 4. Nur Kyrie. – (274v) Aliud. – (276r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ascendens 5. Nur Kyrie. – (276v) Aliud, am Blattrand von späterer Hand ergänzt: De Virginibus 6. – (278r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Descendens 7. Nur Kyrie. – (279v) Aliud, am Blattrand von späterer Hand ergänzt: De Beata Virgine minus 8. – (280r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ad proprias laudes 9. – (281v) Ohne Tit., am Blattrand von späterer Hand ergänzt: De B. M. Virgine maius 10. Gloria mit Tropus Spiritus et alme (Bosse Nr. 23, mit Abweichungen in der Melodie). Tropus mit Papierstreifen überklebt, Gloria auf später eingefügtem Bl. 282bisr–v ohne Tropus wiederholt, urspr. Forts. auf Bl. 283r. – (284r) Pascale, am Blattrand von späterer Graduale Neocellense I 369 Hand ergänzt: Pascale 11. – (285v) De martiribus. Nur Kyrie. – De uno martire sive confessore. – (287r) De virginibus. – (289r) Dominicale. Nur Kyrie. – Ad priorem. 2.2 (291r–311r) Sanctus. Agnus Dei. (291r) Ohne Tit. – (292r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De apostolis 1. – (293r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De virginibus 2. – (294r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De martyribus 3. – (295r) Ohne Tit. Sanctus mit Tropus Benedictus Marie natus qui venit … – (296r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Descendens 4. Sanctus mit Tropus Benedictus Marie filius (?) qui venit … (radiert). – (297r) Ohne Tit. Sanctus mit Tropus Benedictus Marie natus qui venit …, Agnus Dei mit Tropus Summi celi qui tollis … – (298v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Maius de b. virgine 5. Sanctus mit Tropus Benedictus Marie filius qui venit … – (299r) Ohne Tit. – (300r) Ohne Tit. – (301r) Ohne Tit. Sanctus mit Tropus Benedictus Marie filius qui venit … – (301bisr) Ohne Tit., am Blattrand von späterer Hand ergänzt: De confessoribus 6. – (302r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De b. virgine minus 7. – (302v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ad propr. laudes 8. – (303v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Paschale 9. – (304r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Dominicale 10. – (305r) Ohne Tit. – (306v) Ohne Tit. Agnus Dei mit Tropus O dulcis Ihesu Christe ab arce patris misse tu redemisti omnia nostra delesti crimina dona nobis pacem … (Melodie mit M. Schildbach, Das einstimmige Agnus Dei und seine handschriftliche Überlieferung vom 10. bis zum 16. Jahrhun dert, Diss. Erlangen, Nürnberg 1967, 122, Nr. 149 übereinstimmend, jedoch kein Verweis auf Tropen). – (306v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Angelicum 11. – (307v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ascendens 12. – (308v) Ohne Tit. – (309v) Ohne Tit. – (310r) Ohne Tit. – (310v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Simplex 13. 3 (311r–314r) Missa contra pestilentiam. 4 (314v) Antiphonen. – (315v) Karfreitagsliturgie. Trisagion mit Antiphon Crux fidelis omnes … – (316r) Ecce lignum. Beati immaculati. 5 (316v–317r) Alleluia-Verse. (316v) Dnca 5. post pascha. – (317r) In ascensione dni (Dominica 1a ascensionis). 6 (317r–324r) Ergänzung von späterer Hand (Terminus post quem 1457 [Einführung des Festes In transfiguratione dni]) zum Temporale. (317r) In transfiguratione dni. Mit Sequenz (AH 50, Nr. 274). – (319v) De resurrectione. Zwei Sequenzen (AH 54, Nr. 148; AH 50, Nr. 240, unvollständig). – (321v) Expl. mut. (Sequenz, Str. 3a): … superis dies duplicis baptismi legis evan[gelii (2 Bl. fehlen) (324r) Inc. mut. (Sequenz, Str. 6b): ] parenti veteri tu membris forcioribus … – Rubrik: Versus sequens obmissus est supra dominica 8a post penthe. in alleluia. (Text) Replebimur in bonis domus tue sanctum est templum tuum mirabile in equitate. 7 (324v) Sanctus mit Verweis auf vorangehendes Kyriale. 8 (325r) Schlussschrift (rot): Hunc librum comparavit venerabilis pater dominus Nycolaus per missione divina prepositus Novecellum (!) pro suo monasterio ob reverenciam virginis gloriose. Qui incipit regere anno a nativitate domini Mo CCCCo XXXIXo in die concepcionis prefate virginis gloriose. Scriptus et finitus est per fratrem Fridericum Zolner sacerdotem professum eiusdem monasterii. Anno domini millesimo quadringentesimo quadragesimo secundo in vigilia nativitatis eiusdem virginis gloriose. Cui est honor laus et gloria per infinita seculorum secula. Amen (7. September 1442). 370 Graduale Neocellense II Daneben von späterer Hand in Rot, Textualis formata (Textura) ergänzt: dominus Fridericus qui scripsit hunc librum obiit 1446 (!). (325r) Notiz von späterer Hand am rechten Rand der vierten Textzeile in Bastarda: obiit 1449 feria 3a post assumpcionem s. Marie. (Ir–v) leer. G. G. / U. S. ohne Sign. GRADUALE NEOCELLENSE II Perg. I, 317, I Bl. 700 × 500. Neustift, Mitte 15. Jh. (zw. 1442 und 1446). B: Hs. um 1980 restauriert (s. bei G). Perg. am rechten unteren Blattrand tw. stark abgegriffen, daher Ecken vielfach im Zuge der Restaurierung ergänzt bzw. ersetzt. Am Blattrand vereinzelt kleine Löcher (selten auch im Schriftfeld) und Risse, selten Nahtspuren, bisweilen Naht erhalten (Bl. 20, 38, 52, 55, 261). Risse und Fehlstellen mit Perg. überklebt und ergänzt, Bl. 49 fehlende Textstellen nachgetragen. Korrekturen durch Aufkleben von zusätzlichen Pergamentstücken und -streifen (z. B. Bl. 133r, 165r, 187v, 197v, 198v, 199r, 203v, 253r–v, 288r). Lagen: (I–1)I (Vorsatzbl. 20. Jh.) + (IV–1)7 + 6.IV55 + (IV+1)63(64) + 10.IV143(144) + (IV+1)151(153) + 14.IV263(265) + (IV+1)271(274) + V279(284) + IV287(292) + (IV–1+1)295(300) + IV303(308) + (IV+1)312(317) + (I–1)I* (Nachsatzbl. 20. Jh.). Gegenbl. zum Vor- bzw. Nachsatzbl. als Spiegel auf dem VD bzw. HD aufgeklebt. Gegenbl. zu Bl. 7 heraus geschnitten (ohne Textverlust). Bl. 56bis eingehängtes Einzelbl. Gegenbl. zu Bl. 57 und 58 (urspr. Bl. 62 und 63) herausgeschnitten (Textverlust, Reste der Ausstattung im Falz sichtbar), im Zuge der Restaurierung durch leere Bl. ersetzt. Urspr. Gegenbl. zu Bl. 102 und 103 herausgeschnitten, später durch zwei Einzelbl. ergänzt (auf Falz aufgeklebt, in Foliierung berücksichtigt). Bl. 144I eingehängtes Einzelbl. mit (im Kontext) ergänztem Text von späterer Hand (Vierliniensystem etwas enger), in Foliierung nicht berücksichtigt. Bl. 264bis eingehängtes Einzelbl. (Pap., Schrift- und Notenspiegel an die restliche Hs. angepasst) mit Forts. des Gloria von Bl. 264v ohne Tropus (s. bei Inhalt), in Foliierung nicht berücksichtigt. Bl. 295 relativ zeitnah mit fortlaufendem Text ersetzt (ohne rote Foliierung, in Bleistift später ergänzt). Bl. 312 eingehängtes Ein zelbl., auf Bl. 311 aufgeklebt. Bis Bl. 287 (= Ende der Lage 36) am Lagenende Kustoden in römischen Zahlen sowie Reklamanten. Am oberen Blattrand zeitgenössische Foliierung in roten römischen Zahlen I–CCCXII; Fehler in der Zählung: 96 ausgelassen (stattdessen XCVII doppelt angeführt), nach Bl. 274 zwei Blätter ungezählt (heute 274bis und 274ter). S: Schriftraum 535/540 × 350/360, seitlich von zweifachen roten Tintenlinien begrenzt (linke Spalte Noten schlüssel, rechte Spalte Kustos), jeweils sieben rote Vierliniensysteme (39 mm) mit Textzeilen. Zirkelstiche. Textualis formata (Textura), 1442–1446. Hauptschreiber: Friedrich Zollner (vgl. Graduale I, Bl. 325r). Ein schübe von mehreren anderen zeitgenössischen Händen (z. B. Bl. 160r–164v; Bl. 179r, Z. 4, Ergänzung auf Bl. 187v; Bl. 199r, Bl. 203v; Bl. 246v, Z. 1; Bl. 252v, Z. 3–253v, Z. 1; Bl. 272r, Z. 4–276v, Bl. 298r–v, Bl. 304v, Z. 4–312r). Nur vereinzelt Radierungen und Korrekturen (sowohl bei Noten als auch bei Text). Zahlreiche Radierungen, Durchstreichungen, Ergänzungen und Korrekturen von späteren Händen, meist ebenfalls in Textualis formata (Textura), tw. auch in Rotunda (z. B. Bl. 63r, 95r–96v, 253v, 312v). Blattge staltung (Höhe der Vierliniensysteme sowie Schriftgröße) meist nachgeahmt. Zusätze in schwarzer und roter Tinte sowie Bleistift bis ins 18. Jh., z. B. Vacat, Gliederungsstriche von unterschiedlicher Breite, Rubriken, Buchstaben für die Aufführungspraxis (mit Orgel? bzw. Bl. 223r–v, 225r in Rot Chorus – Rectores), Blatt verweise. Mehrere Feste zur Gänze von späterer Hand gestrichen. N: Gotische Choralnotation (mit Virga) auf vier roten Linien, c- bzw. f-Schlüssel, Kustoden. Beginn neuer Abschnitte (z. B. am Beginn des zweiten Teils des Kyriale auf Bl. 275r) vielfach von anderer Hand 15. Jh.: sog. „umgekehrte Hufnagelnotation“. Wie im Graduale I Verwendung von Kustoden in Rot als Hinweis auf einen Schlüsselwechsel innerhalb eines Gesangs. Reperkussionston mit einem ornamentalen Haarstrich. Im 16./17. Jh. zahlreiche Melismen gestrichen und durch Pergamentstreifen überklebt, Notation bisweilen ergänzt bzw. verändert. Wie im Graduale I Ergänzung (18. Jh.) von langen Gliederungsstrichen in Rot (oben lilienförmig bzw. Rhombusform), die über das Vierliniensystem hinausreichen, möglicherweise Hinweis auf Graduale Neocellense II 371 Alternatim-Aufführungsform. Am Blattrand im Kyriale bisweilen Ergänzung von Buchstaben (18. Jh.) als Hinweis zur Aufführungspraxis. A: Rubriken und Foliierung in Rot; in Textpassagen (z. B. Bl. 131v–132v) abwechselnd große Paragraph zeichen in Rot und Blau, tw. mit (interlinearen) Ausläufern und Fibrillen, sowie rote Durchstreichungen; Bl. 281v ein Teil der Notenzeile mit rotem Rautengitter ausgestrichen. Bl. 312v die beiden Anfangsbuchsta ben mit etwas (Besatz-)Dekor. Fleuronnée: fast 1400 Fleuronnée-Initialen (abwechselnd rote und blaue, in der Regel eine Notenzeile hohe Buchstaben mit Punktverdickungen, selten Ausläufer mit Achterschlaufen und Fibrillen; Einleitungsinitialen des Kyriale und der Forts. desselben, Bl. 254r und 275r, in der Höhe von zwei Notenzeilen und, wie die meisten Initialen an den Abschnittsanfängen in diesen Graduale-Teilen und einige Initialen im Commune Sanctorum und im Sanctorale, als aufwändigere rot-blau gespaltene Initialen mit verzierten Blattausspa rungen gestaltet; Bl. 56r, 252v, 272r, 296v, 300v, 304r, 306v, 308r, 309v rot-blau gespaltene Initialen von anderen Initialmalern) und ca. 260 fleuronnierte Cadellen, zumeist in der Höhe einer Notenzeile; von diesen etwa 1460 erhaltenen fleuronnierten Buchstaben das Gros von der Hand des Hauptflorators; FleuronnéeNachträge von mindestens drei weiteren Händen (s. u.). Aufwändigere Fleuronnée-Initialen des Hauptflora tors im Commune Sanctorum: Bl. 13v, 14r, 14v, 15r, 15v, 16r (2), 17r (2), 37r, 37v (2), 38r, 38v, 39r, 50r, 50v, 51r, 51v, [63r], 64r; Sanctorale: Bl. 85v2, 89v, 92v2, 98v, 124r, 125r, 132r, 133v, 135r, 138r, 139r, 148r3, 169r1, 174v, 178r2, 193r, 206r; Kyriale (ohne Einleitungsinitiale): Bl. 254r4, 254v3, 256r1, 256r3, 256v1, 256v4, 257r2, 257r5, 258v1, 258v4, 260r, 260v4, 261r, 262r1, 262v, 263v1, 264r5, 266r1, 266r5, 267v1, 267v5, 267v9, 269r1, 269r8, 270v1, 270v5, 271r; Forts. des Kyriale (ohne Einleitungsinitiale): Bl. 275v2, 276v2, 277v, 278v, 279v, 280r2, 281r, 282r, 283r1, 283v, 284v, 285r, 286r1, 286v, 287r1, 288r, 289r, 290r, 290v2, 291v, 292v2. Fleuronnée der Haupthand wie im ersten Gradualband sehr gleichmäßig und akkurat ausgeführt; wenige Motive, die trotz unterschiedlichster Kombinationen einen relativ gleichförmigen Gesamteindruck erzeu gen. Formenvokabular des Cadellen-Fleuronnée sehr ähnlich, aber noch etwas reduzierter (z. B. weniger Besatz), außer bei Cadellen in den obersten Notenzeilen, die etwas aufwändiger geschmückt sein können (z. B. Bl. 70r, 86v, 144v, 172r, 206v); Cadellen-Fleuronnée in Schwarz, mit wenig rotem Dekor, vorwiegend Konturlinien und Punkte. Blaue Initialen mit Fleuronnée in Rot, (manchmal verblasstem) Violett, seltener in Grün; rote Initialen vorwiegend mit grünem, seltener mit rotem oder violettem Fleuronnée; bei aufwän digeren Initialen Farbkombinationen im Fleuronnée, v. a. grün-violett und grün-rot, bei Einleitungsinitialen grün-violett-rot; wenige Fälle von andersfarbigen, evtl. nachgetragenen Punkten (z. B. Bl. 140v3). Fleuronnée-Motive sehr ähnlich wie im ersten Gradualband mit folgenden Unterschieden: Häufiger gestri chelte Halbpalmetten mit deutlicherer Längungstendenz, insbes. in Binnenfeldern als S-förmig ineinander geschmiegte Paare (z. B. Bl. 8r1), auch in Kombination mit runden Knospen (z. B. Bl. 12v2), diese Knospen zuweilen gepunktet oder statt der Knospen kleine, rundblättrige Blüten (z. B. bei Cadelle Bl. 66r). Als Besatz auch große Halbpalmetten mit gebogten Rändern und plastisch herausschraffiertem Rückengrat (z. B. obere Cadelle Bl. 86v), die den aus den Buchstabenkörpern der aufwändigeren Fleuronnée-Initialen ausgesparten Halbpalmetten stark ähneln. Bei den quadratischen Binnenfeld-Kachelungen werden Kreuzverbindungen zu Stielen, Unterquadrate zu Blättchen (z. B. Bl. 106v1); zudem radial unterteilte Binnenfelder (z. B. Bl. 289r: Kompartimente jeweils mit Knospen gefüllt). Im Cadellen-Fleuronnée häufiger die aus schwarzem Binnen feldgrund ausgesparten ondulierenden Halbpalmettenranken. Bei den typischen Profilköpfen die mürrisch nach unten gezogenen Münder tw. (wie zum Sprechen) seitlich leicht geöffnet (z. B. Bl. 29v, zweite Cadelle) und häufiger „fließende“, an den Verlauf des Buchstabenkörpers angepasste Kinnpartien (z. B. Initiale Bl. 110v; noch ausgeprägter Bl. 74v1, hier auch Fadenausläufer, der direkt dem Mund des Profilkopfs ent wächst). Weitaus mehr aufwändig gestaltete Initialen (ca. 70) mit rot-blau gespaltenen Buchstabenkörpern, die mit binnendekorierten vegetabilen Aussparungen in symmetrischer Anordnung verziert sind (zu Halb palmetten s. o., zudem ondulierende Ranken mit Herz- und Rundblättern, die in Rankenvoluten eingeschrie bene Blüten ausbilden können, z. B. bei Sanctus-Initiale Bl. 275r; hier Blüten auch im Binnenfeld, in eng ondulierender Ranke; Rankenvoluten rot‑, Binnenfeld grüngrundig; Bl. 14v Aussparungen bräunlich-grün ausgemalt); aufwändigere Initialen darüber hinaus mit längeren Fadenausläuferbündeln (rote Bündelstriche, Fäden doppelgestrichelt, symmetrisch angeordnet und mit zurückschwingenden Häkchenenden), die ortho gonal und/oder diagonal abstehen können (z. B. Bl. 281r), sowie mit üppigen Fleuronnée-Leisten, die der Hauptflorator aus den gängigen Motiven aufbaut (z. B. Bl. 267v: hier verbindet die Leiste die drei aufwändi gen Initialen dieser Seite; etwas ausgefallener Bl. 37v: aneinander gereihte Elemente des Laufenden Hunds, denen hängende Herzblätter fibrillenartig entwachsen) und fallweise in Fadenausläuferbündeln enden lässt. Auch die im vorliegenden Gradualband tätige Deckfarbenhand (zu diesem Miniator s. unter „Deckfarben schmuck“ sowie Rischpler) führte etwas Fleuronnée aus: z. B. goldenes Binnenfeld-Fleuronnée der Orna mentalinitiale Bl. 175r (zu weiteren Beispielen dieses Fleuronnée s. erster Gradualband unter „Fleuronnée). 372 Graduale Neocellense II Darüber hinaus mindestens drei (jüngere) Fleuronnée-Hände, vielfach unter Einfluss des Hauptflorators: (1) Bl. 56r–v, 113r (oben), 160r–164v, 272r–274terr, 303r–304v: über 80 Initialen und Cadellen mit variations reichem, sauber gezeichnetem Fleuronnée (Bl. 56r1 aufwändiger); diese Hand auch im ersten Gradualband tätig (Ergänzungshand 1). (2) Bl. 144bisr–v, 304r2 (?), 305r–310v ca. 40 Fleuronnée-Nachträge (Bl. 306v2, 308r2 und 309v aufwändiger: u. a. mehrfarbiges Fleuronnée und längere Ausläufer; unklar ist die Zugehö rigkeit der beiden Fleuronnée-Initialen Bl. 310r–v) von der geübten, wohl in das 1. Fünftel 16. Jh. zu datie renden Hand, die in Cod. 405 (dat. 1501, hier das Fleuronnée flüchtiger), Cod. 194 (dat. 1507), Cod. 142 (Teil mit in Rede stehendem Fleuronnée, dat. 1521) zu finden ist, einige Initialen im ersten Band des ZollnerGraduales gezeichnet und Sign. 14820 zur Gänze ausgestattet hat (ausführlich zu dieser Hand bei Sign. 14820). (3) Bl. 203v, 301v: flüchtig dekorierte Cadellen. Unklar ist auch die Zuordnung der Cadelle Bl. 288v (mit spitznasigem Profilkopf). Deckfarbenschmuck. Ein Wappen; acht (erhaltene) figürliche Initialen in der Höhe von zwei bis drei Noten zeilen (Bl. 1r, 3r, 13r, 49v, 76r, 102v, 112r, 210v) und fünf ornamentale Initialen, jeweils zwei Notenzeilen hoch (Bl. 36v, 142r, 171v, 175r, 186r), jeweils mit fallweise belebten Ranken; ein herausgeschnittenes Folio (Bl. 62) mit figürlichem Deckfarbenschmuck, der von Hermann noch beschrieben wurde, woraus folgt, dass diese Seite nach 1905 entfernt worden sein muss. – Gestaltung der Bestandteile sowohl der figürlichen als auch der ornamentalen Initialen (Rahmen, Außengründe, Buchstabenkörper etc.) unterliegt keiner strikten hierarchischen Gewichtung, d. h. figürliche Initialen wurden nicht wesentlich üppiger angelegt als ornamen tale (Ausnahme: Rankendrolerien, s. u.). Dieser Gradualband war den Neustifter Augustiner Chorherren demnach so kostbar, dass jede Deckfarbenseite annähernd den gleichen Illuminationsaufwand rechtfertigte. Deckfarbenschmuck von einer Hand ausgeführt, die auch Einleitungsseite des ersten Gradualbandes gestal tete; nachfolgend die Charakteristika dieses Illuminators: Initialrahmen eher einfach gehalten, aber zumeist mit andersfarbigem Binnenrähmchen versehen; nur Rah men Bl. 1r (mit Stufenband), 76r (mit feinen Goldranken überzogen) und 36v, 112r, 210v (jeweils mit plas tisch herausgearbeiteten Zierrauten, so auch im ersten Gradualband, Bl. 1r) etwas aufwändiger. Außengrün de bzw. -zwickel in der Regel mit Blattgold belegt, tw. mit rahmenden Linien aus Punktpunzen umgeben und zumeist von feinem, in weißlichem Malmittel ausgeführten Rankenwerk überzogen, seltener mit radial vom Buchstabenkörper ausgehenden Strahlen (Strahlendekor in dieser Technik oft auch auf den vergoldeten Hei ligenscheinen); Bl. 112r Außengrund zartgrün mit goldenen, in Voluten gelegten Blättchenranken. Elegante Buchstabenkörper (mit feinen, plastisch herausgearbeiteten Konturierungen), die in dünne Rankenäste über gehen und seltener mit Stufenbändern, meist mit Blattwerk gefüllt sind: einerseits ondulierend, mit Adern, die mit Reihen von Perlen gefüllt sind, welche sich zur Blattmitte hin vergrößern („Erbsenschotenmotiv“), sowie mit den charakteristischen zweifachen Querstrichen (beide Motive z. B. Bl. 210v), andererseits lang und flach aufliegend, mit mehr oder weniger gebogten Blatträndern (z. B. Bl. 49v). Binnenfelder einfarbig (gerne in Blau) grundiert, oft von feinem goldenem Gitter überzogen, in dessen Quadrate stilisierte vierblätt rige Blüten eingeschrieben wurden (fallweise integrieren sich die mit zarten Maßwerkbögen verzierten Hei ligenscheine der Figuren in dieses Gitternetz, z. B. Bl. 3r), oder von goldenen Blättchenranken, die sich zu Voluten eindrehen und dabei Blüten umschließen können (z. B. Bl. 142r); Bl. 112r blauer, mit Goldsternen übersäter Grund; nur Bl. 175r spitzknospiges, gepunktetes Fleuronnée (in Gold) im orangeroten Binnenfeld; Bl. 1r zusätzlich schlichter gelber Rahmen um das Binnenfeld geführt. Akanthusranken zweiseitig bis (annähernd) umlaufend und auf Rectos von der römischen Foliierung unter brochen, häufig in dichte Voluten gelegt, mit herausgearbeiteten Rippen (immer wieder verlängerte und als Fibrille auslaufende Mittelrippe) und mit sich tw. zurückbiegenden, spitz zulaufenden Einzelbl.; Bl. 142r auf dem linken Blattrand (fast im Falz verschwindend) italianisierender, sich um Vertikalachse windender Rankentyp; seltener Rankenstäbe (Bl. 3r fußt Initiale auf dünnem Stab, von dem aus sich Akanthusäste auf dem Seitenrand verzweigen); Bl. 1r des ersten Gradualbands wächst ein Rankenast zw. Buchstabenkörper und Binnenrahmen hindurch und legt sich hinter den Nimbus Gottvaters; singulär das naturalistische Rosen rankenstück Bl. 210v (rechte untere Ecke); Verzierung durch (Fantasie-)Blüten (zu den Akeleien [Spiel karten-Motive] Bl. 1r s. van Buren, Edmunds, 27 [D.8]), Manschetten, Goldtropfen (in Astgabelungen, tw. einfach punziert und mit Gitterbemalung in hellem Malmittel) mit langen schwarzen Strahlen, Goldpunkte mit kürzeren und längeren, orthogonal abstehenden, schwarzen Fibrillen und seltener Goldfelder, die die Flächen in Rankenvoluten oder zw. sich überschneidenden Ästen ausfüllen; wenig (jetzt nachgedunkelter) Silberdekor, z. B. Rankenfeld neben Initiale Bl. 210v oder Tropfen auf Bas-de-page Bl. 175r. V. a. auf Folios mit historisierten Initialen Ranken mit Tierdrolerien und figürlichen Medaillons geschmückt, s. auch Einlei tungsseite des ersten Gradualbandes; lediglich bei Ornamentalinitiale Bl. 36v Drolerien im Akanthus (s. u. in Auflistung); durch Abreibungen und Beschädigungen des Marginaldekors im Bas-de-page-Bereich und an den unteren Seitenecken evtl. einige Drolerien nicht mehr erkennbar. Figuren fallweise mit anatomischen Unsicherheiten, beispielsweise Bl. 13r ungelenke Knieposition des lin ken Märtyrers (ebenso bei König David, Bl. 1r im ersten Band, kniende Körperhaltung etwas diffus, da Un terschenkel und Füße unter Gewandausläufern nicht deutlich erkennbar sind), Bl. 76r verdrehter Körperbau Graduale Neocellense II 373 des aus dem Boot steigenden Petrus (rechtes, über den Bootsrand gestrecktes Bein sitzt an der Stelle des linken) und Bl. 102v: nacktes Jesuskind ähnelt Gliederpuppe, Positionierung seines linken Ärmchens ist unklar, ebenso die Darstellung der Arme des Priesters (sollten sie sich unter dem Schal befinden, dann hätten sie nur Stumpflänge), zudem ungeschickter Armansatz bei der Marienfigur. Gesichter in der Farbigkeit des Inkarnats, bei Mund- bzw. Lippenformen, Augenschnitten (durchgängig lediglich die deutlich gesetzten, dunklen Pupillen) und Gestaltung des Haupthaars so variationsfreudig, dass sich die Frage stellt, ob diese Komponenten alle von einer Hand stammen oder ob hier nicht Arbeitsteilung anzunehmen ist. Charakte ristisch die betonte Längung von Händen, die zum Gebet gefaltet sind (z. B. bei Märtyrern Bl. 13r). Unbe holfenheiten in der Darstellung der körperlichen Gestalt werden durch geschickte Anlage der Gewanddrape rien und v. a. durch gekonnte Behandlung von Stofflichkeiten aufgewogen: So wirkt z. B. die Bischofskasel (Bl. 49v) mittels breiter grauer Schattenstreifen im Brustbereich schwer und ruhig, der zweifarbige Mantel des hl. Bartholomäus (Bl. 1r) durch den kaskadenartigen Faltenwurf sehr bewegt und das weiße Marien gewand (Bl. 112r) nicht nur durch die reiche Fältelung, sondern auch durch die zartgelben Schattierungen duftig-leicht. Einige Detailfreude bei Stoffmustern (s. z. B. priesterliches Schultertuch Bl. 102v oder den Mantel Davids im ersten Gradualband Bl. 1r), v. a. aber bei der Ausarbeitung von Edelstein‑ und Perlendekor (s. Mitra Bl. 49v, Marienkrone Bl. 210v und Davidskrone, erster Band Bl. 1r) und auch bei der Zeichnung der Holzmaserung (s. z. B. Thron Bl. 210v). Keine Raumdarstellungen oder Architekturen, Figuren agieren vor teppichartigen Ornamentalgründen; Altar Bl. 102v und Betpult Bl. 112r in etwas unsicherer Parallelperspektive, Thron Bl. 210v zentralperspektivisch gegeben (unklare Gestaltung von Sitzfläche und Lehne, die dreieckig wirkt). Klares und leuchtendes Kolorit; charakteristisch das zarte Gelb, das im Zusammenspiel mit Rosa und Weiß einen irisierenden Eindruck erzeugt (z. B. Bl. 112r auf den Spitzen der Füllblätter im rosafarbenen Buchsta benkörper oder als Schattierung des weißen Mariengewands, s. o.) und im Gesichtsinkarnat die Niedertracht der bezeichneten Figur betont (s. Schergen Bl. 13r); das Seelenfigürchen (erster Band Bl. 1r) wird durch wenige Gelbhöhungen auf seinem hellen Gewand zu einer Lichtgestalt; üppiger Einsatz von Blattgold; wenn Goldbelag abgerieben bzw. abgegriffen, rosafarbener Bolus gut erkennbar (s. v. a. Bas-de-page-Zonen); simple Punzierungen (Punkte, kleine Kreise). Bl. 1r Apostel Bartholomäus mit Winzermesser und geschlossenem Buch, vor dem E-Querstrich auf Wie sengrund stehend; in Bas-de-page-Ranken Neustifter Wappen: in blaugrundigem, von feinen, weißen Ran ken überzogenem Vierpass roter Schild mit goldenem Tau-Kreuz; Drolerien: Libelle, zwei Fliegen, Falter, Eichhörnchen (?), hockender Bär mit Dudelsack; grauer, auf einem Bein stehender Vogel mit zurückgebo genem Kopf und aufgestellten Schwanzfedern, dessen langem Schnabel sich kleinerer grüner Vogel nähert; grauer Vogel: Kopie nach Kupferstichmotiv des Spielkartenmeisters: Vogel-Drei, Reiher unten links (s. Lehrs I 110, 61; von Lehrs I 143, explizit im Neustifter Graduale identifiziert). Bl. 3r Apostel Petrus (mit Schlüssel und Buch) und Paulus (mit Schwert und Buch) auf grauem Grund stehend; auf dem linken Seitenrand zwei Rankenmedaillons mit je einem Kopf, der obere blond, der untere schwarzhaarig, im Bas-de-page-Akanthus weiteres Medaillon mit der Büste eines Augustiner Chorherrn im Dreiviertelprofil mit hoher, gewölbter Stirn, markanter Nase, mürrisch nach unten gezogenem Mund, aus geprägtem Kinn mit grauem Bart (?), kapuzenartig hochstehendem Kragen und schwarzer Kappe; Laußer mayer (248) sieht eine Tonsur, was den Schluss nahelegt, dass die Kopfbedeckung nach 1965 nachgetragen wurde (jedoch fraglich). Bl. 13r Enthauptung von Märtyrern: Der Scherge, der gerade einem Märtyrer den Kopf abgeschlagen hat (dieser liegt auf dem Boden), wischt sein Schwert ab; auf grauem Grund kniet ein weiterer Märtyrer und betet; in den Ranken auf dem oberen Seitenrand: Eichhörnchen, auf dem Bas-de-page in zwei Medaillons: Kopf eines vornehmen Mannes mit roter Kappe mit Hermelinrand und Kopf einer Frau, die Krone mit drei Blattzacken trägt (Medaillon stark berieben). Bl. 36v ornamentale Initiale; in den Ranken folgende Drolerien: blaugrüner sowie gelblich-brauner Vogel, an Rankenast angeleinter dunkelgrauer Bär, dunkelgraues Eichhörnchen, zwei grüne Papageien, großer Hase auf separatem Wiesenstück. Bl. 49v Bischof mit prächtiger Mitra, schwarzem geschlossenem Buch und Stab (mit Sudarium) steht auf Wiesengrund. [Bl. 62r Hl. Agathe] Bl. 76r Berufung der ersten Jünger: In einem Boot, das in fischreichem Wasser schwimmt, drei nimbierte Männer, von denen einer schlafend (?) am Ruder sitzt, der zweite (Andreas) ein Netz mit Fischen an Bord zieht und der dritte (Petrus) sich Jesus (mit unbeschriftetem Spruchband) zuwendend aus dem Boot steigt. Bl. 102v Darbringung Christi im Tempel: Maria reicht das Kind über Altar hinweg dem Priester, hinter Maria Frau mit zwei Opfertauben auf der verhüllten Rechten. Bl. 112r Verkündigung: Maria kniet mit vor der Brust verschränkten Armen an hölzernem Betpult, auf dem ein aufgeschlagenes Buch liegt, zu ihren Füßen schlafendes Hündchen, ihr gegenüber der Engel (mit Pfauen flügeln), der mit der Linken einen Lilienzweig hält und mit der Rechten Maria segnet; über ihm zwei mit 374 Graduale Neocellense II Goldbuchstaben beschriftete und mit Zierpunkten geschmückte Spruchbänder (erstes Band: Spiritus sanctus super[ve]niet und Ave gratia plena dominus tecum; zweites Band: Ecce ancilla domini fiat); im oberen Teil des Binnenfelds Büste des segnenden Gottvaters (mit Weltkugel in der Linken) auf stilisiertem Wolkenband, von dem goldene Strahlen ausgehen, auf welchen das nimbierte Jesuskind (mit gelbem Kreuz in Händen) und die Taube des Heiligen Geistes zu Maria hinabgleiten. Bl. 210v Thronende Muttergottes mit Kind wird von Augustiner Chorherrenpropst angebetet; Maria im blauen Sternenmantel mit reich verzierter Krone sitzt auf schlichtem Holzthron, das Kind (ebenfalls in be sterntem Gewand) steht auf ihrem rechten Oberschenkel, hat den linken Arm um ihre Schultern gelegt und greift mit der Rechten nach dem Spruchband (in Goldlettern mit Zierpunkten: O fili dei miserere mei), das sich über dem vor dem Thron knienden, bartlosen Kleriker wölbt; dieser im hellen Chorherren-Mantel mit grauem Kapuzenkragen (mit Schweifen besetztes Feh-Almutium) und dunkler Kappe; Akanthusranken durch verhältnismäßig viele Drolerien belebt: sitzender Bär mit Dudelsack, vier Fliegen, zwei Schnecken, blau-gelber (?) Vogel, Eichhörnchen, grüner Papagei, grüne Eidechse, angeleintes (?) Äffchen mit Rund spiegel, roter Falter und dunkelgrauer Vogel; dieser sitzt auf Rosenzweigvolute, die zartrosa Heckenrose umschließt; daneben in Akanthusvolute Büste eines im Dreiviertelprofil gegebenen braunhaarigen, bärtigen Mannes, der mit blauem Mantel und Chaperon über einem weißen Gewand bekleidet ist (Sendelbinde auf linke Schulter gelegt); unter seinem Kinn weiße Halsbinde sichtbar; mit gefalteten Händen, an denen er helle Handschuhe trägt, hält er Spruchband Der liben Maria nam, vertikal über ihm weiteres Band Servacius Lichtensteyn de Prussia (beide Beschriftungen in Goldbuchstaben). Bildnisse: Die in Rede stehenden Darstellungen sind nicht als (Selbst‑)Porträts im Sinne getreuer Abbilder mit individuellen Zügen zu verstehen, sondern als bildliche Repräsentanten der betreffenden Persönlichkei ten. – 1.) Laußermayer (247) vermutet in der Büste des Augustiner Chorherrn auf dem Bas-de-page Bl. 3r ein „verborgenes Porträt des Miniators“; Hermann (226) sieht darin ein Schreiber‑ oder Miniatorenbildnis. Friedrich Zollner, der Schreiber der Gradualien, ist als Augustiner Chorherr belegt (s. Giner 10); es ist also durchaus denkbar, dass das Medaillon auf Bl. 3r an Zollner erinnern soll, der sich auch im stattlichen Kolo phon (erster Gradualband, Bl. 325r–v) zusammen mit dem Auftraggeber, Propst Nikolaus Scheiber, nennt. Laußermayer sieht – in der Tat nachvollziehbare – physiognomische Ähnlichkeiten (niedrige Stirn, lange Nase mit Wulst über der Wurzel) zw. dem Chorherren-Bildnis Bl. 3r und den Profilköpfen im Fleuronnée (vgl. z. B. im ersten Band, Bl. 31v1; detaillierte Beschreibung der Köpfe s. erster Band unter „Fleuronnée“), was allerdings nicht zu bildnisbezogener Interpretation verleiten sollte. – 2.) Bl. 210v wird der Augustiner Chorherr, der die Madonna anbetet, durch den Fehschweifbesatz seines Almutiums als Propst ausgewiesen. Wie schon Hermann (227) und Laußermayer (248) vermuteten, dürfte er den Auftraggeber, Propst Nikolaus Scheiber, verbildlichen (s. Giner 10 und vgl. Darstellungen im ersten Band, Bl. 227v und Bl. 261r, Bas-depage). – 3.) Zur Darstellung und Person des Servacius Lichtensteyn de Prussia auf Bl. 210v s. Rischpler. S. R. E: Bibliothekseinband: helles Leder über Holz. S. Maria di Rosano (Firenze), Ende 20. Jh. Einband sowie Bindung im Zuge der Restaurierung unter Verwendung von Beschlägen und Schließen unbe kannter Herkunft erneuert. Am VD keine Beschläge erhalten, am HD Eckbeschläge in Form von steigenden gekrönten Löwen mit eckigen Buckeln, in der Mitte runder ornamentaler Beschlag (romanisch?) mit Buckel. Am unteren Rand filigrane Kette, untere und obere Stehkanten (jeweils zwei ca. 5 cm lange Eisenkanten) integriert. Zwei Langriemen-Leder-Metallschließen, obere Schließe schmucklos (möglicherweise original), untere Schließe Renaissance-Löwenkopf mit Datierung 1548 (wohl kein Bezug zur Hs.); am HD aufgena geltes Deckblech (original), hier funktionslos. Kanten gerade. Rücken: sieben Doppelbünde. Kapitale mit naturfarbenem Garn umschlagen. Spiegel sowie Vor- und Nachsatzbl. Perg. 20. Jh., leer bis auf Etikett der Restaurierwerkstätte (s. bei G) am Spiegel des HD. G: Hs. im Auftrag des Propstes Nikolaus Scheiber (Propst 1439–1449) in Kloster Neustift angefertigt. Die zahlreichen Streichungen, Ergänzungen und Hinweise belegen eine Verwendung der Hs. bis ins 19. Jh. Hs. bei vorübergehender Aufhebung des Klosters 1807 nicht an die Universitätsbibliothek Innsbruck gelangt. Am Spiegel des HD Etikett der Restaurierwerkstätte 20. Jh. (Laboratorio di restauro del libro, S. Maria di Rosano [Firenze]), kein Restaurierbericht erhalten. L: Chev. Nr. 28513. – AH 9, Nr. 65. – Hermann 225–227, Nr. 223. – M. Lehrs, Geschichte und kritischer Katalog des deutschen, niederländischen und französischen Kupferstichs im XV. Jahrhundert. 9 Text-, 9 Ta felbände. Wien 1908–1934, hier I, 110 und 143. – B. Rutz, Illuminierte Choralhandschriften zu Neustift. Kir chenmusikalische Jahrbuch 22 (1909) 88–95, hier 90–94. – Laußermayer 246–248, 257–267. – G. Schmidt, Gotik in Österreich. Ausstellung 19. Mai bis 15. Oktober 1967, Minoritenkirche Krems-Stein. Krems an der Donau 31967, 136 (auch in: M. Roland [Hrsg.], Gerhard Schmidt, Malerei der Gotik. Fixpunkte und Aus blicke I: Malerei der Gotik in Mitteleuropa. Graz 2005, 46). – A. H. v. Buren, S. Edmunds, Playing Cards Graduale Neocellense II 375 and Manuscripts: Some Widely Disseminated Fifteenth-Century Model Sheets. The Art Bulletin 56 (1974) 12–30, hier 27. – H. Lehmann-Haupt, Gutenberg and the Master of the Playing Cards. New Haven, London 1966, 35–36, 43. – W. A. Schulze, Alte Dreikönigsbilder in Südtirol. Der Schlern 53 (1979) 5–17, hier 11. – Peintner, Buchmalerei 14, 44, 46–48, 58–62. – M. Peintner, Kloster Neustift. Augustiner Chorherren in Südtirol. Bozen 1985, 19. – N. Rasmo, Kunstschätze Südtirols, Rosenheim 1985, 134–135. – M. H. Peintner, Psallite Deo! Zur Musikgeschichte des Stiftes Neustift, in: 850 Jahre Augustiner Chorherrenstift Neustift 134–153, hier 138. – M. Peintner, Das Skriptorium von Neustift, in: 850 Jahre Neustift. Katalog 91–93, hier 93. – M. Peintner, Musikgeschichte des Stiftes, in: 850 Jahre Neustift. Katalog 94–100, hier 94. – Gozzi 546, Nr. 46. – Neuhauser, Musikgeschichtliche Quellen 151, 161, 164, 194. – Engels 310. – J. G. Alexander, J. H. Marrow, L. Freeman Sandler, The Splendor of the Word. Medieval and Renaissance Illuminated Manuscripts at The New York Public Library. London 2005, hier 181. – M. Studničková, Vážka jako obraz duše hledající spásu. Výzdoba Bratislavského misálu IV (AMB EL 13a) a skupina iluminovaných rukopisů brixenského, vídeňského a jihoněmckého původu z poloviny 15. století / Libelle als Sinnbild der heilsuchenden Seele. Die Ausstattung des Pressburger Missale IV und eine Gruppe illuminierter Handschrif ten aus Brixen, Wien und Süddeutschland aus der Mitte des 15. Jahrhunderts, in: D. Buran (Hrsg.), Galéria. Ročenka Slovenskej národnej galérie / Jahrbuch der Slowakischen Nationalgalerie in Bratislava 2004/2005. Príspevky Medzinárodného kolokvia Gotika na Slovensku a jej stredoeurópsky kontext / Beiträge des Inter nationalen Kolloquiums Gotik in der Slowakei und ihr mitteleuropäischer Kontext. Bratislava 2006, 149– 159, insbes. 158. – Roland, Gotik 274, 288–289 (Kat. 185). – Beier I 15–21, insbes. 16. – Baroffio 325, Nr. 16755. – G. Gabrielli, Tropes in cantus planus sources from South Tyrol, in: J. Border u. a. (Hrsg.), Cantus Planus. Venice meeting 2014. Venezia (in Druck). – Rischpler. Graduale Neocellense. (Ir–v) leer. 1 (1r–76r) Commune Sanctorum. (1r) In vigilia unius apostoli. – (3r) In die sancto. Mit zwei Sequenzen (AH 53, Nr. 228; AH 55, Nr. 6). – (13r) De pluribus martyribus. Mit zwei Sequenzen (AH 53, Nr. 229; AH 55, Nr. 14). – (36r) De uno martyre. Mit Sequenz (AH 55, Nr. 9). – (49v) De uno confessore. Mit drei Sequen zen (AH 54, Nr. 90 auf später eingefügtem Einzelbl. Bl. 56bis; AH 54, Nr. 88; AH 9, Nr. 388). – (62v) De virginibus. Nur Tit. Introitus. (2 Bl. fehlen, Textverlust) (64r) Inc. mut. (Introitus): man]datis tuis que dilexi … Mit Sequenz (AH 50, Nr. 271). 2 (76r–210r) Sanctorale. (76r) In vigilia s. Andreae. – (77r) In die sancto. Mit Sequenz (AH 53, Nr. 122). – (79r) Candidus martyr et pontifex (Rubrik von späterer Hand in Rot durchgestrichen). – Barbara virgo et martyr. Mit Verweis auf Sequenz (AH 50, Nr. 271). Mit Sequenz Gratuletur gloriose unitas nunc fide lium … (nicht in AH, Chev., Mone und Dreves angeführt). Mit Hinweis vacat. – (81r) Nicolaus episcopus et confessor (mit späterer Ergänzung: In eius transactione). Mit Sequenz (AH 55, Nr. 265). – (85r) In octava s. Andreae. – In conceptione BMV (mit Verweis von späterer Hand auf Rasur: Sed Alleluia cum sequencia propria quere in fine libri post Sanctus folio CCCII CCCIII). – Damasius papa (Rubrik von späterer Hand in Rot durchgestrichen). – Lucia virgo et martyr. – Ottilia virgo (Rubrik von späterer Hand in Rot durchgestrichen). – In vigilia s. Thomae. – In die sancto. Mit Verweis auf Sequenz (AH 53, Nr. 228). – (85v) Stephanus. Mit Sequenz (AH 53, Nr. 215). – (89v) Iohannes ev. Mit Sequenz (AH 53, Nr. 168). – (92v) Innocentes. Mit Sequenz (AH 53, Nr. 157). – (95r) Thomas martyr et pontifex. Mit Verweis auf Sequenz (AH 55, Nr. 9). – Silvester papa. Mit Verweis auf Sequenz (AH 54, Nr. 88). – In octava s. Stephani. – In octava s. Iohannis ev. – (95v) In octava Innocentium. – Erhardus episcopus (Rubrik von späterer Hand in Rot durchgestrichen). – Felix presbyter. – Marcellus papa et martyr. – Antonius abbas. – Prisca virgo et martyr. – Fabianus et Sebastianus. – Agnes virgo et martyr. Mit Expl. mut.: … Alleluia. Vers: Felix [ (2 Bl. fehlen, später ersetzt durch zwei eingehängte Einzelbl.) (96r–97v) spätere Ergänzung: (96r) Ignatius. – (97r) In transfiguratione dni. 376 Graduale Neocellense II (98r) Forts. Agnes virgo et martyr. Inc. mut. (Sequenz AH 55, Nr. 51, Str. 12): en]sis cedit morti que vita succedit ecce vidit … – (98r) Vincentius martyr. – (98v) Timotheus ap. – In conversione s. Pauli. Mit Sequenz (AH 50, Nr. 269). – (102v) Policarpus episcopus et martyr. – In octava Agnetis. – Brigida virgo (Rubrik von späterer Hand in Rot durchgestrichen). – Anschließende Rubrik radiert, möglicherweise Ignatius. – (102v) In purificatione BMV. Mit Sequenz (AH 53, Nr. 99). – (106v) Blasius martyr et pontifex. – Agatha virgo et martyr. – (107r) Ingenuinus et Albuinus episcopi. Mit Sequenz (AH 9, Nr. 236). – (110r) Dorothea virgo et martyr. Mit Verweis auf Sequenz (AH 50, Nr. 271). – Amandus episcopus et confessor. – (110v) Scholastica virgo. – Euphrasia virgo (Rubrik von späterer Hand in Rot durchgestrichen). – Euphrosina virgo (Rubrik von späterer Hand in Rot durchgestrichen). – Valentinus martyr. – Faustinus et Iovita (Rubrik von späterer Hand in Rot durchgestrichen). – Iuliana virgo et martyr. – In cathedra s. Petri. – (111v) In vigilia s. Matthiae. – In die sancto. – (am unteren Blattrand von späterer Hand ergänzt:) Iosephus. – (112r) Walpurgis virgo (Rubrik von späterer Hand in Rot durchgestrichen). – Cune gundis virgo. – Gregorius papa. – Gerdrudis virgo (Rubrik von späterer Hand in Rot durchgestri chen). – Benedictus abbas. – In annuntiatione BMV. – (115r) Rupertus episcopus – (am Blattrand von späterer Hand ergänzt:) Conversio Mariae Magdalenae. – Ambrosius episcopus. Mit drei Sequenzen (AH 53, Nr. 36; AH 53, Nr. 50; AH 53, Nr. 67). – (123v) Tiburtius et Valerianus mar tyres. – Georgius martyr. Mit Verweis auf Sequenz (AH 53, Nr. 50). – Marcus ev. Mit Verweis auf Sequenz (radiert, nicht mehr leserlich). – (124r) Trudbertus martyr. – Vitalis martyr. – Philip pus et Iacobus ap. Mit Verweis auf Sequenz (AH 53, Nr. 36). – (125r) (am Blattrand von späterer Hand ergänzt:) Sigismundus. – Inventio s. crucis. Mit Verweis auf Sequenz (AH 54, Nr. 120) und Sequenz (AH 53, Nr. 56). – (128v) Florianus martyr et socii. – Iohannes ante Portam Latinam. Mit Sequenz (AH 55, Nr. 188). – (von späterer Hand ergänzt:) Monica. Mit Verweis auf Sequenz (AH 53, Nr. 50). – (132r) Gordianus et Epimachus. – Pancratius et socii eius. Mit Verweis auf Sequenz (AH 53, Nr. 50). – (132v) In translatione s. Albuini (Rubrik von späterer Hand in Rot durchgestrichen). Mit Verweis auf zwei Sequenzen (AH 53, Nr. 50 und AH 53, Nr. 67). – Victor martyr (Rubrik von späterer Hand in Rot durchgestrichen). Mit Verweis auf Sequenz (AH 53, Nr. 50). – Urbanus papa et martyr. – (133r) Petronella virgo (Rubrik von späterer Hand in Rot durchgestrichen). – Nicomedes martyr (Rubrik von späterer Hand in Rot durchgestrichen). – Marcellinus et Petrus martyres. – Erasmus martyr et pontifex. – Bonifatius et socii eius (Rubrik von späterer Hand in Rot durchgestrichen). – Primus et Felicianus martyres. – Barnabas ap. Mit Verweis auf Sequenz (AH 53, Nr. 228). – Basilides, Cyrinus, Nabor et Nazarius martyres. – Vi tus, Modestus et Crescentia martyres. Mit Verweis auf Sequenz (AH 53, Nr. 229). – Quirinus (Rubrik von späterer Hand in Rot durchgestrichen). – (133v) Marcus et Marcellianus martyres. – Gervasius et Prothasius martyres. – Albanus martyr (Rubrik von späterer Hand in Rot durchge strichen). – Achatius et socii martyres. Mit Verweis auf Sequenz (AH 53, Nr. 229). – In vigilia s. Iohannis Baptistae. – (135r) In die sancto. Mit Sequenz (AH 53, Nr. 163). – (138r) Iohannes et Paulus martyres. – Septem dormientes (Rubrik von späterer Hand in Rot durchgestrichen). – Leo papa (Rubrik von späterer Hand in Rot durchgestrichen). – In vigilia s. Petri et Pauli ap. – (139r) In die sancto. Mit Sequenz (AH 53, Nr. 210). – (141v) In commemoratione s. Pauli. Mit Verweis auf Sequenz (AH 53, Nr. 228). – In octava s. Iohannis Baptistae. – Erentrudis virgo. – In visi tatione BMV. Mit Sequenz (AH 54, Nr. 193). – (144r) Prosula zum Offertorium Filiae regum Mater Christi veneranda (AH 48, Nr. 399; Hofmann-Brandt Nr. 403). Mit Expl. mut.: … De tua letemur prole virgo pre[ (Forts. Bl. 145r) (144Ir–v) spätere Ergänzung von anderer Hand: Forts. bzw. Abschluss der Prosula. Anschließend Alleluia: Hoc alleluia per octavam canitur. (145r) Forts. von Bl. 144v. Inc. mut. (Prosula zum Offertorium): prae]clarissima Amen. – In translatione s. Martini. – Processus et Martinianus martyres. – Udalricus episcopus. – In octava apostolorum s. Petri et Pauli. Mit Verweis auf Sequenz (AH 53, Nr. 228). – Willibaldus epis Graduale Neocellense II 377 copus. – Kilianus et socii eius. – In octava visitationis BMV. – In translatione s. Nicolai. Mit Sequenz (AH 54, Nr. 66). – (148r) Septem fratres. – (148v) In translatione s. Benedicti abbatis. – Margaretha virgo et martyr. Mit Sequenz (AH 54, Nr. 64). – (151v) Hainricus confessor (Rubrik von späterer Hand in Rot durchgestrichen). – In divisione apostolorum. Mit Sequenz (AH 50, Nr. 267). – (155v) Alexius confessor. – Praxedis virgo. – Maria Magdalena. Mit Sequenz (AH 50, Nr. 268). – (159r) Apollinaris martyr et pontifex. – (159v) Christina virgo (Rubrik von späterer Hand in Rot durchgestrichen). – In vigilia s. Iacobi. – In die sancto. Mit Verweis auf Sequenz (AH 53, Nr. 228). – Christophorus martyr (Rubrik von späterer Hand in Rot durchgestrichen). – Anna. Mit Sequenz (AH 55, Nr. 62). – (161v) Martha. Mit Sequenz (AH 55, Nr. 244). – (164v) De virginibus. – (165r) Panthaleon martyr. – Felix papa. – Abdon et Sennen martyres. – Tertulli nus martyr. Si sollempnizatur die dedicacionis Brixinensis cantetur sequentia Spe mercedis (Rubrik von späterer Hand in Rot durchgestrichen). – Petrus ad vincula. Mit Verweis auf Sequenz (AH 53, Nr. 228). – Stephanus papa et martyr. – In inventione s. Stephani. – Oswaldus rex et martyr. – (165v) Sixtus et socii eius. – (am Blattrand von späterer Hand ergänzt:) De transfigura cione domini quere de tempore in fine libri. – Donatus martyr et pontifex. – Afra et sodales eius. Mit Sequenz (AH 53, Nr. 119). – (168r) Cyriacus et socii eius. – (168v) Romanus martyr (Rubrik von späterer Hand in Rot durchgestrichen). – In vigilia s. Laurentii. – (169r) In die sancto. Mit Sequenz (AH 53, Nr. 173). – (171v) Tiburtius martyr. Mit Verweis auf Sequenz (AH 53, Nr. 229). – Hyppolitus et socii eius. – Cassianus martyr et pontifex. Mit Verweis auf Sequenz (AH 54, Nr. 89). – Eusebius confessor. – In vigilia assumptionis BMV. – In die sancto. Introitus Gaudeamus mit Verweis auf Tropus Quia virgo, von späterer Hand Incipit gestrichen und non canitur nach Rasur ergänzt. Mit Sequenz (AH 53, Nr. 104). – (174v) In octava s. Laurentii. – Agapitus martyr. – (175r) Magnus martyr (Rubrik von späterer Hand in Rot durchgestrichen). – Bernardus abbas (Rubrik von späterer Hand in Rot durchgestrichen). – In octava assumptionis BMV. – Timotheus et Symphorianus martyres. – In vigilia s. Bartholomaei. – In die sancto. Mit Verweis auf Sequenz (AH 53, Nr. 228). – Genesius martyr (Rubrik von späterer Hand in Rot durchgestrichen). – Ruffus et Abundus martyres (Rubrik von späterer Hand in Rot durch gestrichen). – Hermes. Mit Verweis auf Sequenz (AH 53, Nr. 229). – Augustinus. Introitus Statuit ei mit Verweis auf Tropus Augustino, von späterer Hand Incipit gestrichen und non dicitur nach Rasur ergänzt. Mit Sequenz (AH 53, Nr. 126). – (178r) In decollatione s. Iohannis Baptistae. – (179r) Iohannes Baptista. Mit Sequenz (AH 50, Nr. 270). – (182r) Felix et Adauctus martyres. – Aegidius abbas. – (183r) In octava s. Augustini. Mit Verweis auf Sequenz (AH 54, Nr. 88) und Sequenz (AH 54, Nr. 32). – (von späterer Hand ergänzt:) In translatione s. Erndrudis. – (186r) Magnus abbas (Rubrik von späterer Hand in Rot durchgestrichen). – Ioachim et Anna (Rubrik von späterer Hand in Rot durchgestrichen). – Regina virgo et martyr. – Adrianus martyr. – In nativitate BMV. Introitus Gaudeamus mit Verweis auf Tropus Nata summa, von späterer Hand Incipit gestrichen und non canitur nach Rasur ergänzt. Mit Sequenz (AH 53, Nr. 95). – (187v) In translatione s. Cunegundis. Mit Verweis auf Sequenz (AH 50, Nr. 271). – Gorgonius martyr. – (von späterer Hand ergänzt:) Infra octavas nativitatis beate Marie. – (188r) Prothus et Iacinthus. – In exaltatione s. crucis. Mit Sequenz (AH 54, Nr. 120). – (192v) In octava BMV. – Nicomedes martyr. – Euphemia virgo. – Lucius et Geminianus (Rubrik von späterer Hand in Rot durch gestrichen). – Lampertus martyr et pontifex. – In vigilia s. Matthaei ap. et ev. – In die sancto. Mit Verweis auf Sequenz (AH 55, Nr. 6). – (193r) Mauritius et socii eius. – In translatione s. Ruperti. Mit Verweis auf Sequenz (AH 54, Nr. 89). – In translatione s. Virgilii. – Cosmas et Damianus. – Wenzelaus dux et martyr (Rubrik von späterer Hand in Rot durchgestrichen). – Michael. Mit Sequenz (AH 53, Nr. 192). – (197v) Hieronymus presbyter. Mit Verweis auf Sequenz (AH 54, Nr. 90). – Remigius, Germanus et Vedastus episcopi (Rubrik von späterer Hand in Rot durch gestrichen). – Leodegarius martyr. – Franciscus confessor. – Sergius et Bacchus (Rubrik von späterer Hand in Rot durchgestrichen). – Marcus papa. – In translatione s. Wolfgangi (Rubrik von späterer Hand in Rot durchgestrichen). – In translatione s. Erhardi (Rubrik von späterer Hand in 378 Graduale Neocellense II Rot durchgestrichen). – Dionysius et socii eius. Mit Verweis auf Sequenz (gestrichen). – Gereon et socii eius. – In translatione s. Augustini. Mit Verweis auf Sequenz (AH 54, Nr. 32). – (198r) Maximilianus episcopus. – Colomannus martyr. – Calixtus papa et martyr. – Gallus abbas. – In translatione s. Marthae. – Lucas ev. Mit Verweis auf Sequenz (AH 55, Nr. 6). – (198v) Ianuarius et socii eius. – Ursula et undecim milia virginum. Mit Verweis auf Sequenz (AH 55, Nr. 333). – Severus episcopus (Rubrik von späterer Hand in Rot durchgestrichen). – Severinus episcopus. – Crispinus et Crispinianus martyres. – Amandus episcopus. – In vigilia s. Simonis et Iudae. – In die sancto. Mit Verweis auf Sequenz (AH 53, Nr. 228). – (199r) Narcissus martyr et episcopus. – (am Blattrand von späterer Hand ergänzt:) Wolfgangus episcopus. – In vigilia omnium sanctorum. – In die sancto. Mit Sequenz (AH 53, Nr. 112). – (200v) Eustachius et socii eius. – Leonhardus confessor. – Willibrordus episcopus. – In octava omnium sanctorum. – Quattuor coronati. – Theodorus martyr. – Martinus episcopus. Mit Sequenz (AH 53, Nr. 181). – (203r) Brictius epis copus. – (203v) am Blattrand später ergänzt: Leopoldus. Mit Verweis auf Sequenz (AH 54, Nr. 90). – Othmar abbas. – In octava s. Martini. Mit Verweis auf Sequenz (AH 9, Nr. 388). – Elisabeth. Mit Sequenz (AH 55, Nr. 120). Alleluia auf Rasur. – (206r) Caecilia virgo et martyr. – Clemens papa et martyr. – (206v) Chrysogonus martyr. – Catharina virgo et martyr. Mit Sequenz (AH 54, Nr. 56). – (210r) Virgilius episcopus. Mit Verweis auf Sequenz (AH 54, Nr. 88). 3 (210r–253r) De commemoratione BMV. (210r–243v) Propriumsgesänge mit 13 Sequenzen: (212r) AH 54, Nr. 219. – (214r) AH 50, Nr. 241. – (217v) AH 54, Nr. 227. – (219v) AH 54, Nr. 248. – (222r) AH 54, Nr. 225. – (223v) AH 54, Nr. 237. – (225r) AH 54, Nr. 218. – (227r) AH 42, Nr. 74 (Str. 6 zw. Str. 1 und 2). – (229v) AH 54, Nr. 238. – (232v) AH 54, Nr. 232. – (234v) AH 54, Nr. 245. – (238v) AH 54, Nr. 221. – (240v) AH 54, Nr. 279. – (242r) Offertorium Recordare mit Prosula (AH 49, Nr. 634). – (243v) In adventu dni de BMV. Mit Sequenz (AH 54, Nr. 191). – (246r) Infra nativitatem usque ad purificationem. Mit Verweis auf zwei Sequenzen (AH 54, Nr. 2 und AH 53, Nr. 15, Str. 9–12) und Sequenz mit Hinweis vacat (AH 53, Nr. 15, bei Beginn der Str. 9 am Blattrand Verweis von späterer Hand Hic de beata v.). – (248r) De BMV post festum purificationis usque ad Quadra ges. – De BMV in Quadrages. diebus sabbatis post primas feriales. – (248v) Tempore paschali de BMV. Mit drei Sequenzen (AH 9, Nr. 65, mit Nennung der Hs. bzw. Chev. Nr. 28513, mit Nen nung der Hs.; AH 54, Nr. 18 und AH 54, Nr. 21). – (252v) von anderer Hand: Post ascensionem. Mit Sequenz (AH 54, Nr. 226). (253v) Offertorium von späterer Hand 17./18. Jh. 4 4.1 (254r–293r) Kyriale. (254r–272r) Kyrie. Gloria. (254r) Ohne Tit. Nur Kyrie. – Aliud, am Blattrand von späterer Hand ergänzt: De Apostolis 1. – (256r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De Confessoribus 2. Nur Kyrie. – (256v) Aliud Kyrie, am Blattrand von späterer Hand ergänzt: Angelicum 3. – Aliud, am Blattrand von späterer Hand ergänzt: De martiribus 4. Nur Kyrie. – (257r) Aliud. – (258v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ascendens 5. Nur Kyrie. – Aliud, am Blattrand von späterer Hand ergänzt: De virginibus 6. – (260r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Descendens 7. Nur Kyrie. – (260v) Aliud, am Blattrand von späterer Hand ergänzt: Minus de B. V. Maria 8. – (262r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ad proprias laudes 9. – (263v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Maius de B. Virgine 10. Mit späterer Abänderung: Bl. 264bisr–v (Pap.) später eingefügt, urspr. Version auf Bl. 265r fortgesetzt mit Tropus Spiritus et alme (Bosse Nr. 23, mit Abweichungen in der Melodie). – (266r) Pascale, am Blattrand von späterer Hand ergänzt: Paschale 11. – (267v) De martiribus, am Blattrand von späterer Hand ergänzt: 12. Nur Kyrie. – De uno martire sive confessore, am Blattrand von spä terer Hand ergänzt: 13. – (269r) De simplicibus virginibus, am Blattrand von späterer Hand Graduale Neocellense II 379 ergänzt: 14. – (270v) Dominicis diebus, am Blattrand von späterer Hand ergänzt: 15. Nur Kyrie. – Ad priorem, am Blattrand von späterer Hand ergänzt: 16. 5 (272r–274v) Alleluia-Rufe mit Sequenz. (272r) De s. confessore [In die translationis] mit Zuweisung an Rudbertus, von späterer Hand an Virgilius). Sequenz (AH 54, Nr. 89). – (274v) De s. Cassiano. Sequenz (AH 54, Nr. 89 [Text leicht abweichend]). 4.2 (274terr–v) Zusatz zum Kyriale. (274terr) In inventione crucis. Offertorium. – (274terv) Kyrie. Später ergänzt: 20. 4.3 (275r–293r) Forts. des Kyriale: Sanctus. Agnus dei. (275r) Ohne Tit. – (275v) Ohne Tit., am Blattrand von späterer Hand ergänzt: De Apostolis 1. – (276v) Ergänzung am Blattrand fast zur Gänze weggeschnitten: ]ini 2. – (277v) Ohne Tit., am Blattrand von späterer Hand ergänzt: De martyribus 3. – (278v) Ohne Tit., am Blattrand von späterer Hand ergänzt: 13. Sanctus mit Tropus Benedictus Marie natus qui venit … – (279v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Descendens 4. Sanctus mit Tropus Bene dictus Marie filius qui venit … – (280r) Ohne Tit. Sanctus mit Tropus Benedictus Marie natus qui venit. .., Agnus Dei mit Tropus Summi celi qui tollis … – (281r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Maius de Beata Virgine. Sanctus mit Tropus Benedictus Marie filius qui venit … (in Rot durchgestrichen). – (282r) Ohne Tit. – (283r) Ohne Tit. – (283v) Ohne Tit. Sanctus mit Tropus Benedictus Marie filius qui venit … – (284v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Minus de Bea. Virgine 6. – (285r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ad proprias laudes 7. – (286r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Paschale 8. – (286v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Dominicale 9. – (287v) Ohne Tit. – (288r) Ohne Tit., am Blattrand von späterer Hand ergänzt: De Confessoribus 10. Agnus Dei mit Tropus O dulcis Iesu Christe, radiert und nur Forts. auf Bl. 289r lesbar: ]misse tu redemisti … – (289r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Angelicum 11. – (290r) Ohne Tit., am Blattrand von späterer Hand ergänzt: Ascendens 12. – (290v) Ohne Tit. – (291v) Ohne Tit. – (292r) Ohne Tit. – (292v) Ohne Tit., am Blattrand von späterer Hand ergänzt: Sim plex 13. 5 (293v–304r) Forts. der Alleluia-Rufe mit Sequenz (BMV). (293v) Die ersten zwei Notenlinien leer, dann Alleluia und Sequenz (AH 54, Nr. 201). – (296v) AH 54, Nr. 278. – (300r) AH 54, Nr. 250. – (302r) AH 54, Nr. 188. 6 (304r–312r) Sequenzen für Heilige und BMV. (304r) Henricus (AH 55, Nr. 159). – (306v) Achatius et socii (AH 55, Nr. 38). – (308r) Sequen cia de beata virgine post nativitatem domini (AH 54, Nr. 2). – (309v) Ursula et undecim milia virginum (AH 55, Nr. 333). 7 (312v) Ergänzung von späterer Hand 18. Jh.: Introitus. Minuisti eum paulo minus ab Angelis gloria et honore corona … (Ir–v) leer. G. G. / U. S. ANHANG 383 Verzeichnis der heute noch an der ULB Tirol befindlichen, zu Beginn des 19. Jahrhunderts aus Neustift dorthin gebrachten Handschriften Die 1921 nicht an Italien abgetretenen Handschriften sind in den Katalogbänden zu den Handschriften der Universitäts- und Landesbibliothek Tirol in Innsbruck mit genauen Beschreibungen berücksichtigt. Hier erfolgt lediglich ein Überblick (Stand 2021). Cod. 59 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. I, 380 Bl. 318 × 215. Erfurt, um 1455. Cod. 82 HENRICUS DE LANGENSTEIN Pap. I, 37, I Bl. 285 × 207. Wien (?), 15. Jh. Cod. 157 ARISTOTELES, LAT. Pap. I, 55, I* Bl. 285 × 223. Wien, 1366. Cod. 159 ALBERTUS DE SAXONIA Pap. I, 73, I* Bl. 285 × 224. Wien, 1365. Cod. 164 ANSHELMUS HAVELBURGENSIS Pap. I, 37, I* Bl. 287 × 213. Tirol (?), Ende 14./Anfang 15. Jh. Cod. 169 SCHWABENSPIEGEL Pap. I, 130, I* Bl. 285 × 207. Neustift (?), um 1410–1420. Cod. 170 ALANUS AB INSULIS Pap. I, 131, I* Bl. 275 × 203. Neustift (?), ca. 1460. Cod. 172 IOHANNES DE SOMMERFELD Pap. I, 142, I* Bl. 277 × 199. Neustift (?), um 1425. Cod. 175 SERMONES CONCILII BASILIENSIS Pap. IV, 235 Bl. 283 × 203/206. Rheinland (?), um 1440. 384 Handschriften an der Universitäts- und Landesbibliothek Tirol Cod. 176 PROTOKOLL DES REICHSKAMMERGERICHTES SPEYER, 1522–1533, LAT./DT. Pap. I, 329, I* Bl. 285/290 × 205. Speyer, 1522–1533. Cod. 421 THOMAS EBENDORFER Pap. I, 428, I* Bl. 310 × 210. Süddeutschland (?), 3. Viertel 15. Jh. Cod. 553 NEUSTIFTER RITUALE (FRAGMENT) Pap. I, 25, I* Bl. 335 × 220. Neustift, Anfang 17. Jh. Cod. 593 RECHENBEHELFE UND MESSGERÄT Pap. I, 24, I* Bl. 265 × 185. Entstehungsort unbestimmt, 1443. Cod. 596 (KIRCHEN-)HISTORISCHE SAMMELHANDSCHRIFT Pap. 38 Bl. 280 × 205/210. Entstehungsort unbestimmt, 15. Jh. (T. II: Ende 15. Jh.). Cod. 598 THEOLOGISCHE SAMMELHANDSCHRIFT, U. A. IOHANNES CAPISTRANUS Pap. 22 Bl. 265 × 205. Sachsen oder Brixen (?), 2. H. 15. Jh. Cod. 642 FORMELSAMMLUNG FÜR RECHTSGESCHÄFTE Pap. I, 53, I* Bl. 220 × 160. Brixen, 1. H. 16. Jh. Cod. 681 RHETORISCHE SAMMELHANDSCHRIFT (U. A. JOHANNES WILDENAUER) Pap. I, 115 Bl. 205 × 150. Südtirol (?), 1. H. 16. Jh. Cod. 692 AENEAS SILVIUS PICCOLOMINI (PIUS II). (PS.?-)FRANCISCUS DE ASSISI Pap. 4 Bl. 225 × 155. Entstehungsort unbestimmt, Ende 15. Jh. (nach 1480). Cod. 693 IACOBUS FISCHER Pap. 14 Bl. 210 × 150/155. Neustift, 1588. Cod. 717 MECHTHILD DE HACKEBORN. PS.-BONAVENTURA Pap. 20 Bl. 160 × 105. Entstehungsort unbestimmt, 2. H. 15. Jh. Cod. 718 SERMO DE FLAGELLIS DEI Pap. 12 Bl. 160 × 105. Neustift, 16. Jh. Cod. 723 MELCHIAS UKEN Pap. I, 39, I* Bl. 155 × 100. Entstehungsort unbestimmt, nach 1773. Cod. 762 BREVIARIUM BRIXINENSE Pap. IV, 201 Bl. 215 × 140. Brixen (?), 9. Jz. 15. Jh. Handschriften an der Universitäts- und Landesbibliothek Tirol Cod. 763 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 313 Bl. 213 × 140/145. Neustift (?), 15. Jh. Cod. 764 SERMONES DE TEMPORE. LEGENDEN Pap. I, 371 Bl. 215 × 145. Kärnten (?), 15. Jh. Cod. 765 EVANGELISTAR, DT. Pap. 196 Bl. 220 × 159. Tirol, 1472. Cod. 788 GELDSCHULD ZU NATZ (URBAR), DT. Pap. VII, 132, I* Bl. 210 × 155. Südtirol, 1. H. 16. Jh. (u. a. 1510). Cod. 799 COMPUTUS. ARS PUNCTANDI. COLUTIUS SALUTATUS Pap. 18 Bl. 205 × 150. Südtirol (?), um 1530. Cod. 800 SENSATUS Pap. 24 Bl. 280 × 195. Entstehungsort unbestimmt, Mitte 15. Jh. Cod. 842 SCHWABENSPIEGEL. AUSZUG AUS DEM LANDRECHT, DT. Perg. I, 33, I* Bl. 205 × 145. Entstehungsort unbestimmt, 2. H. 14. Jh. Cod. 922 SPIEGEL DEUTSCHER LEUTE (DEUTSCHENSPIEGEL), DT. Perg. II, 80, I* Bl. 220 × 170. Süddeutschland (Neustift?), 14. Jh. Cod. 960 NEUSTIFTER-INNSBRUCKER SPIELHANDSCHRIFT, DT. Pap. I, 60, I* Bl. 275/280 × 105/110. Schmalkalden oder Neustift, 1391. Cod. 961 HANS VINTLER Pap. 3 Hefte (40, 21 und 28 Bl.) 305 × 110. Tirol, um 1415. Cod. 988 AUGUSTINER CHORHERRENSTIFT GRIES (BOZEN): STIFTUNGEN UND ÄBTELISTE Pap. III, 15 Bl. 160 × 105. Gries (Bozen), 1651. Cod. 992 DECIMUS IUNIUS IUVENALIS Pap. I, 49, I* Bl. 305 × 210. Entstehungsort unbestimmt, 4. Viertel 15. Jh. Cod. 1013 AUCTARIUM GRAMMATICAE Pap. 14 Bl. 195 × 145. Neustift (?), 1. H. 16. Jh. Cod. 1014 AENEAS SILVIUS PICCOLOMINI (PIUS II.) Pap. 4 Bl. 215 × 150. Neustift, 15. Jh. 385 386 Handschriften an der Universitäts- und Landesbibliothek Tirol Cod. 1015 KOSMOGRAPHIE Pap. 32 Bl. 205 × 155. Neustift, 1509. Cod. 1031 HIERONYMUS I. PIESENDORFER Pap. I, 43 Bl. 160 × 110. Neustift, 1535–1536. Cod. 1190 KANONIKERLISTE DES CHORHERRENSTIFTS NEUSTIFT Pap. 2 Bl. 245 × 200. Neustift, 1815. Urspr. Besitzer bzw. Zwischenbesitzer (nicht Letztbesitzer) Neustift Diese Handschriften befanden sich einst in Neustifter Besitz, sie sind jedoch nicht direkt aus Neustift an die ULB Tirol gelangt. Cod. 457 SERMONES. CANTIONARIUM. Perg. 107 Bl. 240 × 170. Tirol (?), 14. Jh. bzw. Neustift (?), Ende 14./Anfang 15. Jh. (Letztbesitzer: Schnals) Ink. 156 F 20 BREVIARIUM SALISBURGENSE (INK.) MIT HANDSCHRIFTLICHEM ANHANG FÜR NEUSTIFT Perg. 12 Bl. 165 × 115. Neustift, 1487 und 1501. (Letztbesitzer: Innsbruck, Wappenturm-Bibl.) Fraglich, ob aus Neustift an die ULB Tirol gelangt Cod. 15 MISSALE BRIXINENSE Perg. 232 Bl. 395 × 295. Augsburg oder Tirol, um 1490. Cod. 21 ANTIPHONALE Perg./Pap. 224 Bl. 369 × 255. Halltal, 1492. Cod. 47 FRONLEICHNAMSPREDIGT, DT. Pap. 7 Bl. 160 × 109. Tirol, 16. Jh. Cod. 96 ABBILDUNGEN VON PFLANZEN UND TIEREN Perg./Pap. I, 31, I* Bl. 437 × 350. Italien, 18. Jh. Cod. 111 PIRRO ANTONIO FERRARO. GIOVANNI BATTISTA FERRARO Pap. 2 Bde. ca. 335 × 220. Italien, 17. Jh. Handschriften an der Universitäts- und Landesbibliothek Tirol 387 Cod. 189 BARTHOLOMAEUS A SANCTO CONCORDIO Pap. I, 203, I* Bl. 316 × 225. Oberitalien, 1433. Cod. 233 IOHANNES HEROLT („DISCIPULUS“) Pap. I, 241 Bl. 285 × 215. Tirol (?), 1439. Cod. 321 PONTIFICALE BZW. RITUALE (BENEDICTIONALE) Perg. II, 36 Bl. 195 × 140. Tirol, 1497. Cod. 351 BREVIARIUM Perg. V, 322 Bl. 180 × 125. Schlesien (Diözese Breslau), 14. Jh. Cod. 370 BENEDICTIONALE UND RITUALE Perg. 47 Bl. 163 × 115. Süddeutschland (Bamberg oder Salzburg?) und Halltal, 15. Jh. (wohl nach 1481). Cod. 387 RAIMUNDUS DE PENNAFORTE Perg. IV, 348, IV* Bl. 135/140 × 95. Frankreich, 14. Jh. Cod. 394 BREVIARIUM Pap. 330 Bl. 145 × 105. Hauptteil Schlesien, Zusätze Österreich, wohl Tirol, vor 1512 (um 1465/1468). Cod. 442 COLLECTIO CONCLUSIONUM ET RESOLUTIONUM SACRAE CONGREGATIONIS CARDINALIUM S. CONCILII TRIDENTINI Pap. II, 254 Bl. 260 × 190. Rom (?), Anfang 17. Jh. Cod. 511 BERICHTE DER JESUITENMISSION IN SÜDDEUTSCHLAND UND TIROL Pap. II, 92 Bl. 198 × 159. Tirol, nach 1719. Cod. 563 MISSALE BRIXINENSE („TEYSER-MISSALE“) Perg. VIII, 210 Bl. 295 × 215. Neustift oder Pustertal (Bruneck oder Kiens?), 1414. Cod. 607 IOHANNES DE AUERBACH Pap. 9 Bl. 215 × 145. Diözese Passau, Ende 15. Jh. Cod. 610 BREVIARIUM BRIXINENSE Pap. I, 380, I* Bl. 215 × 150. Neustift oder Brixen, 2. H. 15. Jh. Cod. 616 THEOLOGISCHE SAMMELHANDSCHRIFT Pap. 240 Bl. 215 × 155. Tirol, 1457–1458. Cod. 628 BREVIARIUM PATAVIENSE, PARS AESTIVALIS Pap. 417 Bl. 225 × 135. Diözese Passau, 1486. 388 Handschriften an der Universitäts- und Landesbibliothek Tirol Cod. 632 BREVIARIUM PATAVIENSE (TEILBREVIER) Pap. 202 Bl. 213 × 145. Diözese Passau, Ende 15. Jh. Cod. 640 ANTONIUS AZARO DE PARMA Pap. 348 Bl. 215 × 145. Entstehungsort unbestimmt (Wien?), 1. H. 15. Jh. Cod. 645 ARZNEIBUCH, DT. Perg. 175 × 124, 65 Bl. (130 S.) 175 × 124. Tirol, 16. Jh. Cod. 657 BIBLIA LATINA Perg. II, 402, III* Bl. 185 × 125. Raum Paris, Ende 13./Anfang 14. Jh. Cod. 688 LECTURA INSTITUTIONUM IUSTINIANI Pap. 75 Bl. 200 × 150. Neustift (?), 16. Jh. Cod. 716 BELLUM BOICO-SUEVICUM Pap. I, 25 Bl. 175 × 120. Entstehungsort unbestimmt, 1. H. 18. Jh. Cod. 719 ADAM WALASSER Pap. 123 Bl. 149 × 90. Entstehungsort unbestimmt, 1638. Cod. 727 DE LA GEOMETRIE, FRZ. Pap. I, 147, I* Bl. 104 × 70. Entstehungsort unbestimmt, nach 1624. Cod. 731 STUNDENBUCH, LAT. Pap. 142 Bl. 142 × 105. Entstehungsort unbestimmt, 5. Jz. 15. Jh. Cod. 732 ANDREAS ORNITHOPARCHUS. IOHANNES COCHLAEUS Pap. 171 Bl. 154 × 110. Entstehungsort unbestimmt, 1523. Cod. 783 SAMMELHANDSCHRIFT VON BREVIERTEXTEN (ORDO BREVIARII, OFFICIA, LEGENDEN) MIT AUTOGRAPHEN IOHANNES HINDERBACHS Pap. III, 116 Bl. 215 × 145. U. a. Trient, 2. H. 15. Jh. Cod. 794 NATURWISSENSCHAFTLICHE SAMMELHANDSCHRIFT Pap. I, 53, I* Bl. 200 × 145. Entstehungsort unbestimmt, Anfang 16. Jh. Cod. 818 STATUS CAMERALIS, CIVILIS ET PROVINCIALIS DER GEFÜRSTETEN GRAFFSCHAFFT TYROL, DT. Pap. I, 54, I* Bl. 340 × 235. Tirol, 18. Jh. Cod. 829 LANDRECHT FÜR DIE GERICHTE MÖLTEN, SARNTAL, RITTEN UND VILLANDERS, DT. Pap. I, 61 Bl. 330 × 205. Tirol, 18. Jh. Handschriften an der Universitäts- und Landesbibliothek Tirol Cod. 835 ROGER SCHRANZHOFER, DT. Pap. II, 48, I* Bl. 265 × 210. Mais (Meran) bzw. Lana, 1796 bzw. 1797). Cod. 850 ANTON ROSCHMANN Pap. 31 Bl. 325 × 215. Innsbruck, 1742. Cod. 958 PETRUS CANTOR Perg. I, 24, I* Bl. 285 × 195. Entstehungsort unbestimmt, 14. Jh. Cod. 962 SAMMELHANDSCHRIFT Pap. 174 Bl. 210 × 145. Paris (?), Mitte 15. Jh. (u. a. Paris 1457, 1460, 1462, 1466). 389 390 Konkordanz der Signaturen der Stiftsbibliothek Neustift mit den Projektnummern der Hill Monastic Manuscript Library Cod. 7 29.646 Cod. 163 29.654 Cod. 19 29.647 Cod. 165 29.655 Cod. 20 29.695 Cod. 166 29.633 Cod. 31 29.648 Cod. 167 29.688 Cod. 38 29.649 Cod. 168 29.688 Cod. 52 29.650 Cod. 171 29.656 Cod. 53 29.651 Cod. 173 29.688 Cod. 54* 29.668 Cod. 174 29.675 Cod. 56* 29.663 Cod. 177 29.657 Cod. 77 29.652 Cod. 181 29.658 Cod. 78 29.664 Cod. 182 29.658 Cod. 83 29.665 Cod. 183 29.658 Cod. 85 29.666 Cod. 190 29.659 Cod. 86 29.667 Cod. 194 29.637 Cod. 99 29.645 Cod. 196 29.633 Cod. 100 29.632 Cod. 200 29.660 Cod. 106 29.667 Cod. 204 29.661 Cod. 107 29.668 Cod. 217 29.677 Cod. 123 29.653 Cod. 252 29.637 Cod. 125 29.669 Cod. 253 29.677 Cod. 142 29.638 Cod. 282 29.678 Cod. 145 29.662 Cod. 293 29.678 Cod. 146 29.670 Cod. 308 29.679 Cod. 153 29.671 Cod. 315 29.678 Cod. 155 29.672 Cod. 327 29.681 Cod. 156 29.673 Cod. 353 29.635 Cod. 158 29.673 Cod. 360 29.681 Cod. 160 29.654 Cod. 397 29.681 Cod. 161 29.654 Cod. 405 29.636 Cod. 162 29.654 Cod. 440 29.676 Konkordanz Signaturen Stiftsbibliothek Neustift – Projektnummern Hill Monastic Manuscript Library 391 Cod. 447 29.680 Cod. 726 --- Cod. 453 29.685 Cod. 780 Cod. 483 29.690 Cod. 836a* 29.692 Cod. 485 29.690 Cod. 836b* 29.642 Cod. 495 29.689 Cod. 839* 29.696 Cod. 501* 29.642 Cod. 924* --- Cod. 503* --- Cod. 927* --- Cod. 532 29.697 Cod. 930* --- Cod. 550 29.682 Cod. 931* 29.694 Cod. 578 --- Cod. 1012 --- --- Cod. 579 --- Cod. 597 29.674 Sign. 14820 29.639 Cod. 605 --- Sign. 15063 --- Cod. 606 --- Cod. 614 29.691 Nr. 139 (olim Sign. 14819) --- Cod. 617 29.687 Nr. 940 --- Cod. 654 29.634 Cod. 691 --- Neustifter Wappenbuch --- Cod. 694 --- Graduale Neocellense I und II --- Mit * gekennzeichnete Signaturen bezeichnen Archivhandschriften, die im vorliegenden Katalog nicht berücksichtigt wurden. 392 Verzeichnis der Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Die Anordnung der Heiligenfeste folgt jener im zweiten Band des Graduale Neocellense, der das umfassendste Sanctorale der beschriebenen Handschriften aufweist. Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Missalien, Gradualien Cod. 100 Cod. 154 Andreas Nr. 139 Nr. 940 ● ● Breviere, Antiphonarien GN II ●, * Candidus pontifex ● Barbara ● Nicolaus In conceptione BMV ● ● ● ● Lucia ● ● Thomas ●* Stephanus ● Iohannes ev. ● Innocentes ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● ● Erhardus episcopus ● Felix presbyter ● Marcellus papa ● Antonius abbas ● Prisca ● ● ● ● ● ● ● ● ● ● ● Thomas pontifex Fabianus et Sebastianus ● ● Silvester papa Vincentius Nr. 940 ● Ottilia Thimotheus ● Nr. 139 ● Damasus papa Agnes Cod. 405 Sign. 15063 ● ● ● ● ● ● In conversione s. Pauli ● Policarpus ● Brigida ● ● 393 Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Missalien, Gradualien Cod. 100 Cod. 154 Nr. 139 Nr. 940 Ignatius Breviere, Antiphonarien GN II In purificatione BMV ● ● Agatha ● Ingenuinus et Albuinus episcopi ● Dorothea ● Amandus episcopus ● Scholastica ● Euphrasia ● Euphrosina ● Valentinus ● Faustinus et Iovita ● Iuliana ● In cathedra s. Petri ● Matthias Nr. 940 ● ● ● ● ● ● ● ● ● ● ● ● ●* Iosephus ● Walpurgis ● Cunegundis ● Gregorius ● Gertrudis ● Benedictus abbas ● ● ● ● ● Rupertus episcopus ● ● ● Conversio Mariae Magdalenae ● ● ● Ambrosius ● Tiburtius et Valerianus ● Georgius ● Marcus ev. ● Trudbertus ● Vitalis Philippus et Iacobus ap. Nr. 139 ● Blasius pontifex In annuntiatione BMV Cod. 405 Sign. 15063 ● ● Sigismundus ● Inventio s. crucis ● ● ● ● ● ● ● ● De lancea dni ● ● ● ● Florianus et socii ● Alexander et Eventius ● Iohannes ante Portam Latinam ● ● ● 394 Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Monica Missalien, Gradualien Cod. 100 Cod. 154 Nr. 139 Nr. 940 ● Breviere, Antiphonarien GN II ● Gordianus et Epimachus ● ● Pancratius et socii ● ● Maria ad martyres ● Translatio s. Albuini ● ● Victor ● ● Urbanus papa ● Nr. 940 ● ● ● ● ● ● Nicomedes ● ● Marcellinus et Petrus ● ● Erasmus martyr et pontifex ● ● Bonifatius et socii ● ● Primus et Felicianus ● ● Barnabas ap. ● ● Basilides, Cyrinus, Nabor et Nazarius ● ● Vitus, Modestus et Crescentia ● ● Quirinus ● Marcus et Marcellianus ● ● Gervasius et Prothasius ● ● ● ● Albanus ● Iohannes Baptista ● ● ●* , Iohannes et Paulus ● Septem dormientes ● Leo papa ● ● ●** ● Petrus et Paulus ap. ● ● ●, * In commemoratione s. Pauli ● ● ● Erentrudis In visitatione BMV Nr. 139 ● Petronella Achatius et socii Cod. 405 Sign. 15063 ● ● ● ● ● ● ● Processus et Martinianus ● Translatio s. Martini ● Udalricus ● Willibaldus episcopus ● Kilianus et socii ● Translatio s. Nicolai ● Translatio s. Benedicti abbatis ● Septem fratres ● 395 Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Missalien, Gradualien Cod. 100 Margaretha Cod. 154 Nr. 139 Nr. 940 ● Breviere, Antiphonarien GN II ● Hainricus confessor ● In divisione apostolorum ● Alexius ● Praxedis ● Maria Magdalena Cod. 405 Sign. 15063 ● ● ● Apollinaris ● Christina ● Iacobus ●* Christophorus ● ● Anna ● ● ● ● De virginibus ● ● Felix papa ● Abdon et Sennen ● Tertullinus ● Petrus ad vincula ● Stephanus papa ● In inventione s. Stephani ● Oswaldus rex ● In transfiguratione dni ● ● ● ● ● ● ● ● Donatus ● Afra et sodales ● Cyriacus et socii ● Romanus ● Laurentius ● ● ●* , ** Tiburtius ● Hyppolitus et socii ● Cassianus martyr et pontifex ● Eusebius ● In assumptione BMV ● ● Sixtus et socii Agapitus Nr. 940 ● Martha Panthaleon Nr. 139 ● ●* ● Magnus ● Bernardus abbas ● Timotheus et Symphorianus ● Bartholomaeus ●* ●* ● ● ● ● ● ● ● ● 396 Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Missalien, Gradualien Cod. 100 Cod. 154 Nr. 139 Nr. 940 Genesius Breviere, Antiphonarien GN II Rufus et Abundus Nr. 940 ● ● ● ● ● ● ● Augustinus ● ● In decollatione s. Iohannis Baptistae ● Iohannes Baptista ● Felix et Adauctus ● Aegidius abbas ● Translatio s. Erentrudis ● Magnus abbas ● Ioachim et Anna ● Regina ● Adrianus ● ● In nativitate BMV ● ● Corbinianus ● Translatio s. Cunegundis ● Gorgonius ● Prothus et Iacinthus ● In exaltatione s. crucis ● Nicomedes martyr ● Euphemia ● Lucius et Geminianus ● Lampertus ● ● ● ● ●* ● ●** ● ●* ● Mauritius et socii ● ●* Translatio s. Ruperti ● Translatio s. Virgilii ● Cosmas et Damianus ● Wenzeslaus Michael archangelus Nr. 139 ● Hermes Matthaeus Cod. 405 Sign. 15063 ●* ● ● ● ● Hieronymus presbyter ● Remigius, Germanus et Vedastus episcopi ● Leodegarius ● Franciscus ● Sergius et Bachus ● Marcus papa ● Translatio s. Wolfgangi ● Translatio s. Erhardi ● ● ● 397 Heiligenfeste in den Missalien, Gradualien, Antiphonarien und Brevieren Heiligenfest * mit Vigil, mit Oktav, * nur Suffragien Missalien, Gradualien Cod. 100 Cod. 154 Nr. 139 Nr. 940 Dionysius et socii Breviere, Antiphonarien GN II Gereon et socii ● ● Maximilianus episcopus ● Colomannus ● Calixtus papa ● Gallus abbas ● Translatio s. Marthae ● Lucas ev. ● Ianuarius et socii ● ● ● ● Severinus episcopus ● ● Crispinus et Crispinianus ● ● Translatio s. Amandi ● Amandus ● Simon et Iuda ●* Narcissus episcopus ● Wolfgangus episcopus ● Dedicatio ecclesiae ● Omnes sancti ● Caesarius ● ● ● ● ● Eustachius et socii ●* , ● ● ● ● ● ● ● ● ● ● ● ● ● Leonhardus ● Willibrordus episcopus ● Quattuor coronati ● Theodorus ● ● ● ● Brictius episcopus ● Leopoldus ● Othmar abbas Elisabeth ● ● ● ● ●* Caecilia ● ● Clemens papa ● ● Chrysogonus Catharina Virgilius episcopus Nr. 940 ● ● ● Severus episcopus Martinus episcopus Nr. 139 ● Translatio s. Augustini Ursula et undecim milia virginum Cod. 405 Sign. 15063 ● ● ● ● ● ● ● 398 Verzeichnis der Hymnen 1. Temporale Incipit Nachweis Anlass Cod. 405 Sign. 15063 A patre unigenitus ad nos venit … AH 2, Nr. 107 Per octavam [epiphaniae dni] ad nocturnum 140r A solis ortus cardine … AH 2, Nr. 23, mit Einschub zw. Str. 3 und 4: AH 27, Nr. 82, Str. 4 In secundis vesperis 137v Ad coenam agni providi … AH 51, Nr. 83 Ad completorium 143r Aeterna Christi munera … AH 52, Nr. 7 Ad completorium 144r Agnoscat omne saeculum … AH 50, Nr. 71 In nativitate dni. Ad nocturnum 137r Audi benigne conditor … AH 51, Nr. 54 Ad nocturnum 141r Beata nobis gaudia … AH 2, Nr. 51 Ad nocturnum 146v Chorus nove Ierusalem novam … AH 50, Nr. 215 Ad laudes 143v Christe qui lux es et dies … AH 51, Nr. 22 Ad completorium 141r Christe redemptor omnium … AH 2, Nr. 22 Ad laudes 137v Christus ascendens choros angelorum … AH 52, Nr. 11 In secundis vesperis 145r Clarum decus ieiunii monstratur … AH 51, Nr. 57 Ad laudes 141v Conditor alme siderum … AH 51, Nr. 47 In adventu dni (Ad vesperas) 135v Corde natus ex parentibus … AH 50, Nr. 26 Ad completorium 138r Dies absoluti praetereunt dies … AH 52, Nr. 2 Dnca in Septuages. 140r Ex more docti mistico … AH 51, Nr. 55 Dnca 1. in Quadrages. 140v Festum nunc celebre … AH 50, Nr. 143 In ascensione dni 145v Gratuletur omnis caro nato … AH 50, Nr. 145, Str. 1–4 und 14 Ad laudes 139v Hostis Herodes impie Christum … AH 50, Nr. 53, Str. 8–9, 11 und 13 In epiphania dni 139v Iam lucis orto sidere … AH 52, Nr. 10 Ad primam 145r Iesu quadragenarie dicatur … AH 51, Nr. 58 Dnca 3. in Quadrages. Ad vesperas 141v Iesum nostra redempto amor et desiderium … AH 51, Nr. 89 Ad completorium et laudes 146r O salutaris hostia … AH 50, Nr. 388, Str. 5 und 6 Ad completorium 148r Pange lingua gloriosae lanceae … AH 52, Nr. 6 De lancea domini ad vesperas 143v Pange lingua gloriosi … AH 50, Nr. 386 In festo corporis Christi ad vesperas 146v Paschali iubilo iunctra sint … AH 52, Nr. 8 Ad nocturnum 144r Praesepe iam fulget … AH 50, Nr. 8, Str. 8 In die natalis (!) domini. Ad vesperas 136v 138v 399 Verzeichnis der Hymnen Incipit Nachweis Anlass Cod. 405 Sign. 15063 Rex Christe factor omnium redemptor … AH 51, Nr. 72 Ad laudes 142v Sacris solennis iuncta … AH 50, Nr. 387 Ad nocturnum 147r Summo parenti gloria natoque … AH 12, Nr. 184, Str. 6 In epiphania dni 139v Texilla regis prodeunt … AH 2, Nr. 42, o. Str. 2 In passione dni 142r Veni creator spiritus mentes … AH 50, Nr. 144 mit Einschub zw. Str. 5. und 6: Da gaudiorum premia da graciarum munera, dissolve litis vincula astringe pacis federa In festo penthecostes 146r Veni redemptor gentium … AH 50, Nr. 8, Str. 2–7 Ad completorium 136r Verbum supernum prodiens … AH 51, Nr. 48 Ad nocturnum 136v Verbum supernum prodiens … AH 52, Nr. 9 Ad laudes 144v Verbum supernum prodiens … AH 50, Nr. 388, Str. 1–4 Ad laudes 147v Vita sanctorum decus angelorum … AH 51, Nr. 85 Tempore paschali 142v Vox clara ecce intonat … AH 51, Nr. 49 Ad laudes 136v 2. Sanctorale Incipit Nachweis Anlass Cod. 405 * nur Verweis Alma mater generosa doctoris … AH 4, Nr. 397 De sancta Monica matre Augustini 151v Anni reduxit circulus festum … AH 4, Nr. 13 Ad nocturnum 157r Assunt Annae solennia huius dent … AH 52, Nr. 105 Anna 155r Assunt festa iubilea in Mariae nunc gaudia … AH 48, Nr. 402 In visitatione BMV 153v Aurea luce et decore … AH 51, Nr. 188 Petrus et Paulus 153r Ave Catherina martyr et regina … AH 52, Nr. 248 Catharina 161v Ave maris stella Dei mater alma … AH 51, Nr. 123 In annuntiatione BMV 150r Ave vitae vitis avia Christi … AH 4, Nr. 141 Ad secundas vesperas 155v Caelestes gemmae confessione nitide AH 51, Nr. 107 cernite … Ad laudes 150r Caeli cives applaudite … Chev. I Nr. 3471 Ad nocturnum 158v Caelum digne tripudiat terra … AH 23, Nr. 139 Ingenuinus et Albuinus 149v Christe caelorum habitator … AH 51, Nr. 104 Ad nocturnam 162v Christe cunctorum dominator … AH 51, Nr. 103, Str. 1–6 Ad laudes et secundas vesperas 163r Christe redemptor omnium conserva … AH 2, Nr. 81 Ad laudes 160v Christe sanctorum decus angelorum … AH 50, Nr. 146 Michael 159v Concentu parali hic te Maria … AH 53, Nr. 99 In purificatione BMV. Si fuerit infra Septuagesimam in secundis vesperis pro ymno dicitur sequens 149r Conscendant usque sidera … AH 50, Nr. 227 Laurentius 157v De patre verbum prodiens … AH 51, Nr. 161 Iohannes ev. 138v Nr. 940 400 Verzeichnis der Hymnen Incipit Nachweis Anlass Cod. 405 Nr. 940 * nur Verweis De sacro tabernaculo … AH 50, Nr. 44 Ad nocturnum 154r En miranda prodigia concepit … AH 48, Nr. 404 Ad laudes 154v Exorta a Bethsaida duo … AH 52, Nr. 99 Andreas 148r Eya fratres extollamus … AH 52, Nr. 335 In deposicione s. Ruperti 150v Fit porta Christi pernia referta plena … AH 27, Nr. 1081, Str. 4–6 Ad completorium et laudes 150v Gaude visceribus mater … AH 51, Nr. 125 In nativitate BMV 158v Haec sacras Iesu lacrimis plantas … AH 51, Nr. 175, Str. 3 und 4 Ad laudes 155r Hymnizet nunc plebs … AH 4, Nr. 380 Martha 156r Hymnum Deo vox iocunda … AH 52, Nr. 181, Str. 1–4 und 10 Ad nocturnum 161r Iesu Christe auctor vitae * AH 51, Nr. 174 Afra 155r Iesu Christi auctor vitae * AH 51, Nr. 174 De sancta Affra 157v Iesu Christi auctor vitae … AH 51, Nr. 174 Maria Magdalena 154v Iesu salvator saeculi redemptis … AH 51, Nr. 130 Ad nocturnum 160r Laetetur plebs fidelium … AH 19, Nr. 391 Ad laudes 152r Magnae laetitiae fulget lux hodie … AH 4, Nr. 381 In secundis vesperis 156r Magne pater Augustine preces … AH 52, Nr. 117 Augustinus 158r Martine confessor Dei valens … AH 27, Nr. 154 Martinus 160v Novum sidus emicuit … AH 52, Nr. 182 Elisabeth 161r O laudat laetitia lux hodierna … AH 51, Nr. 183 Nicolaus 148v O mater Christi fulgida … AH 48, Nr. 403 Ad completorium 154r O nata lux de lumine … AH 51, Nr. 99, Str. 1–4, 6–7 In transfiguratione dni 157r O sancta mundi domina … AH 51, Nr. 122 In nativitate BMV 159r O stella maris fulgida absolve … AH 52, Nr. 74 Ad nocturnum 155v Omnes superni ordines … AH 50, Nr. 228 Omnes sancti 160r Patris nostri Augustini mater sancta Monica … AH 4, Nr. 398 Ad nocturnum 151v Petre pontifex inclite Christi … AH 51, Nr. 190 Petrus ad vincula 156v Quem terra pontus aethera … AH 50, Nr. 72, Str. 1–2, 4–8 In assumptione BMV 157v Quod chorus vatum venerandus … AH 2, Nr. 28 In purificatione BMV 148v Salve crux sancta salve mundi gloria … AH 50, Nr. 223 In inventione s. crucis 151r (mit Divisio) Sancte Dei preciose protomartyr Stephane … AH 48, Nr. 79 Stephanus 138v Solennis dies advenit qua virgo … AH 51, Nr. 160 Ad nocturnum 139r Tandem viro destituta … AH 52, Nr. 181, Str. 7–9 Ad laudes 161v Tibi Christe splendor … AH 50, Nr. 156 Ad laudes et secundas vesperas 159v Urbs beata Ierusalem … AH 51, Nr. 102 In dedicatione ecclesiae 162r Ut queant laxis resonare … AH 50, Nr. 96 Iohannes Baptista 152v Votiva cunctis orbita lucis triumphat … AH 51, Nr. 175, Str. 1, 2 und 5 Ad nocturnum 155r 227v (mit Divisio) 401 Verzeichnis der Hymnen 3. Commune Sanctorum Incipit Nachweis Anlass Cod. 405 Aeterna Christi munera … AH 50, Nr. 17, Str. 1, 3–5 und 8 Ad matutinas 170v Deus tuorum militum sors … AH 51, Nr. 114a In festo unius martiris 169Iv Exsultet caelum laudibus … AH 51, Nr. 108 In festis apostolorum 163v Hic est verus Christi … AH 52, Nr. 70 De simplici confessore (= De uno confessore non pontifice). Ad laudes 188r Iesu corona virginum … AH 50, Nr. 21 De virgine et martyre 191r Iesu redemptor omnium … AH 51, Nr. 117 Ad laudes 179v Iste confessor domini sacratus … AH 51, Nr. 118 De confessore et pontifice 176Iv Martyr Dei qui unicum patris … AH 51, Nr. 113 Ad laudes 172Ir Ortu phebi iam primo … AH 51, Nr. 109 De apostolis ad laudes 167r Rex gloriose martyrum corona … AH 51, Nr. 112 Ad laudes 173r Sanctorum meritis inclita … AH 50, Nr. 153 De pluribus martyribus 169v Virginis proles opiferque … AH 51, Nr. 121 Ad laudes 194r Nr. 940 229v 4. Cursus BMV Nachweis Anlass Cod. 353 Adesto sancta trinitas par splendor una Deitas … Incipit Chev. Nr. 487. AH 51, Nr. 96 De sancta trinitate (Matutin, Vesper) 116r 127v Audi benigne conditor nostras preces … AH 51, Nr. 54 Pro peccatis (Matutin, Laudes) Ave manna caelicum verius legabi … Chev. Nr. 23597 De corpore Christi (Prim) 61r Ave maris stella Dei mater alma … AH 2, Nr. 29 Cursus communis de BMV (Vesper) 157r Ave maris* AH 51, Nr. 123 BMV (Vesper) Ave plenum gaudium vita beatorum pauperum … Chev. Nr. 23749 De corpore Christi (Terz) 62v Ave vivens hostia veritas et vita … AH 50, Nr. 395 De corpore Christi (Laudes) 60r Beate nobis gaudia anni reduxit orbita … Chev. Nr. 2340 De sancto spiritu (Laudes) 28r Castae parentis viscera poena torquentur … Mone II 142, Nr. 439 De compassione BMV (Vesper) 110v Christe qui lux es et dies noctis tenebras detegis … AH 51, Nr. 22 Pro peccatis (Komplet) 31r De omnibus sanctis (Matutin, Vesper) 1r 12v Christe redemptor omnium conserva AH 51, Nr. 129 tuos famulos … Christe sanctorum decus angelorum rector humani generis et auctor … AH 50, Nr. 146 De proprio angelo (alle Tagzeiten) Cod. 405 Cod. 654 16r 20r 214v 2r 4v 5v 7r 8r 9r 10v 11v 402 Incipit Christi felix anima sacrari tuo vultui … Verzeichnis der Hymnen Nachweis AH 30, Nr. 8 Anlass De anima Christi (alle Tagzeiten) Cod. 353 Cod. 405 Cod. 654 184r 186v 187v 188v 190r 191r 193r 195r AH 30, Nr. 81 Pro defunctis 34r 39r 40r 41v 44r 46r 48v 50r Dicamus omnes cernui clamemus atque singuli ploremus … AH 51, Nr. 55 Pro peccatis (Prim, Terz, Sext, Non) 21r 23v 25v 27v Fit porta Christi pervia referta plena gratia … Chev. Nr. 6346 Cursus communis de BMV (Komplet) 160v Hic Iesu veraciter duplex est natura … Chev. Nr. 7807 De corpore Christi (Sext) 64r Hoc ardoris calculo veni nos ignire … Chev. Nr. 27686 De corpore Christi (Vesper) 67r Iesu nate de virgine ex mistico spiramine … Chev. Nr. 9578 De compassione BMV (Prim) 102v Iesu nostra redemptio amor et desiderium Deus creator … AH 51, Nr. 89 Pro peccatis (Vesper) 30r Iesu salvator saeculi redemptis ope subveni … AH 51, Nr. 130 De omnibus sanctis (Laudes, Prim, Terz, Sext, Non, Komplet) 5v 8v 7v 9r 10r 14v Iesum dulcissime cibus salutaris … Chev. Nr. 28563 De corpore Christi (Komplet) 69r De aeterna sapientia (alle Tagzeiten) 13r 16v 18r 19r 20v 21v 23r 24v De compassione BMV (Matutin) 94v Deus aeterne gloriae pater misericordiae … Iesus dulcis memoria dans vera cordis gaudia … Mone I 329 Nr. 258 Imperatrix clementiae rorem infunde Chev. Nr. 8483 gratiae … Novum salutis gaudium caelesti sponsae mittitur … AH 30, Nr. 52 De septem gaudiis Mariae virginis (alle Tagzeiten) 165r 170v 172v 174v 176r 177v 179v 181v Nunc rex creator omnium crucem fers … Chev. Nr. 12570 De compassione BMV (Terz) 104v 403 Verzeichnis der Hymnen Incipit Nachweis Anlass Cod. 353 O gloriosa domina quae tua cruciamina … AH 4, Nr. 87 De compassione BMV (Laudes) O gloriosa femina excelsa super sidera … AH 2, Nr. 27, Str. 5–7 BMV (Laudes) O lux beata trinitas et principalis unitas … Chev. Nr. 13150. AH 51, Nr. 40 und 2, Nr. 17 De sancta trinitate (Prim, Terz, Sext, Non) 121r 123r 124r 125r De misericordia dni (alle Tagzeiten) 40r 44v 46r 47r 48v 49v 51v 52v O lux misericordiae tu primo mortis tempore … Chev. Nr. 30585. AH 30, Nr. 12 O paterne gloriae splendor per respectum tuum … Chev. Nr. 13381. AH 31, Nr. 5 De sancta trinitate (Laudes) Patris sapientia veritas divina … AH 30, Nr. 13 Chev. Nr. 16347. Quem terra pontus aethera colunt … AH 50, Nr. 72 100r 210r 119v 211r 212r 213r 213v 214r 215r 215v De passione Christi (alle Tagzeiten) Cursus communis de BMV (Matutin, Laudes) 131r 141v Qui pressura mortis dura soluisti nexus … AH 50, Nr. 382 De passione Christi (alle Tagzeiten) 71v 76r 78r 80r 82v 84v 88v 91v Rector nunc piissimus ut reus crucifigitur … Chev. Nr. 17058 De compassione BMV (Sext) 106r Regi regum soli vita decus et imperium trinitati … Chev. Nr. 10968. AH 52, Nr. 117, 5 De sancta trinitate (Komplet) 129r De compassione BMV (Non) 107v 143v 147r 149v 152r Rerum Deus tenax vigor Christus in cruce moritur sed poena fit acerbior cum reus non compatitur Rex Christe clementissime tu corda nostra posside … AH 32, Nr. 130 Cursus communis de BMV (Prim, Terz, Sext, Non) Rex Christe clementissime tu corda nostra posside … AH 32, Nr. 130 BMV (Prim, Terz, Sext, Non, Komplet) Sumptum non consumitur corpus salvatoris … Chev. Nr. 33769 De corpore Christi (Non) 65r De compassione BMV (Komplet) 113r Tu lucis ante terminum Christe sepulcro clauderis te plorat virgo virginum Deus humani generis … Cod. 405 211v 213r 214r Cod. 654 404 Incipit Verzeichnis der Hymnen Anlass Cod. 353 De sancto spiritu (Prim, Terz, Sext, Non, Vesper, Komplet) 31v 33r 34v 35v 37r 38v Veni sancte spiritus et emitte caelitus AH 54, Nr. 153 lucis tuae radium … De sancto spiritu (Matutin) 26r Verbum supernum prodiens nec patris linquens … De corpore Christi (Matutin) 55r Veni creator spiritus mentes tuorum visita … Nachweis AH 50, Nr. 144 AH 50, Nr. 388 Cod. 405 Cod. 654 405 Verzeichnis der Reimoffizien Incipit Nachweis Anlass Sign. 15063 Ave gemma virtuosa Dorothea … AH 5, Nr. 56 Deus ante luciferum in sinu sui patris … AH 5, Nr. 13. Cantus Nr. 200447 De conceptione BMV 184r Gaude pia plebs iustorum … AH 5, Nr. 7 De lancea dni 138v Gloria tibi trinitas aequalis una Deitas … in AH nicht angeführt, Beginn mit Chev. Nr. 7287 übereinstimmend In festo ss. trinitatis 110v Gratulemur regi digna psallentes … AH 25, Nr. 42 Barbara 60r Gregorius ut creditur divinitus sic dicitur … AH 5, Nr. 64 Gregorius 78r Inclita sanctae virginis … AH 18, Nr. 37 und AH 26, Nr. 69 Catharina 175v Laetare Germania claro felix germine … AH 25, Nr. 90 Elisabeth 172v O Margareta caelorum virgo secreta … AH 28, Nr. 3 Margaretha 148v O rex altissime Deus mirabilis … zwei rhythmisierte Antiphone: 1. ähnlich wie AH 25, Nr. 85 (Dominicus), 2. ähnlich wie Cantus Nr. 206563 (Monica) In translatione s. Augustini 168v Summae trinitati simplici Deo una divinitas … LMLO XT31 In festo ss. trinitatis Videtur miraculum mater domini in AH nicht angeführt concepit … Dorothea In purificatione BMV Nr. 139 Nr. 940 196r 195r 188r 178r 73v 67v 406 Verzeichnis der Sequenzen 1. Temporale Incipit Nachweis Anlass Cod. 100 Cod. 154 Nr. 139 Nr. 940 30v 27v GN I * nur Verweis, ** unvollständig (Inc. oder Expl. mut. bzw. Fehlstelle) Agni paschalis esu … AH 53, Nr. 50 Dnca resurrectionis dni Agni paschalis esu … AH 53, Nr. 50 Fer. 3 169r Benedicta semper sancta AH 53, Nr. 81 sit trinitas … In festo ss. trinitatis Carmen suo dilecto … AH 53, Nr. 54 Fer. 6 Concentu parili … AH 53, Nr. 99 In purificatione BMV Eia recolamus laudibus … AH 53, Nr. 16 In aurora Eia recolamus laudibus … AH 53, Nr. 16 In nativitate dni Eia recolamus laudibus … AH 53, Nr. 16 In circumcisione dni 7va 22v Festa Christi … AH 53, Nr. 29 In epiphania dni 8vb 25r Gaude Dei genitrix * AH 53, Nr. 15, Str. 9 In circumcisione dni Grates nunc omnes … AH 53, Nr. 10 In galli cantu 3va 8r Grates nunc omnes reddamus * AH 53, Nr. 10 Dnca 1. post nativitatem dni 33r Grates salvatori … AH 53, Nr. 52 Fer. 4 172v Haec est sancta sollemnitas … AH 53, Nr. 56 Sabbato 183v Hodiernae festum lucis … AH 54, Nr. 140 De lancea dni 189v Lauda Sion salvatorem lauda ducem et pastorem … AH 50, Nr. 385 In festo ss. corporis Christi 70vb 2r 179v 10va 24r 12v 4v 34v 19r 37v** 15v 3r 21r 72ra Laudes Deo concinat … AH 53, Nr. 53 Fer. 5 176r Laudes salvatori voce … AH 53, Nr. 36 Dnca resurrectionis dni Mundi renovatio * AH 54, Nr. 148 Dnca 3. post pascha 194r Mundi renovatio * AH 54, Nr. 148 Dnca 4. post pascha 195v Mundi renovatio nova … AH 54, Nr. 148 Dnca resurrectionis dni 62rb 1r 158v 31v 28v 407 Verzeichnis der Sequenzen Incipit Nachweis Anlass Cod. 100 Cod. 154 Nr. 139 Nr. 940 GN I * nur Verweis, ** unvollständig (Inc. oder Expl. mut. bzw. Fehlstelle) 9v (mit Divisio) Natus ante saecula … AH 53, Nr. 15 In aurora Natus ante saecula … AH 53, Nr. 15 In nativitate dni (Ad summam missam) Natus ante saecula … AH 53, Nr. 15 In circumcisione dni Pangamus creatori … AH 53, Nr. 46 Fer. 2 6ra 29r 16r (mit Divisio) 164v Rex regum * AH 50, Nr. 240 Dnca 3. post pascha 194r Rex regum * AH 50, Nr. 240 Dnca 5. post pascha 196v Sancti spiritus adsit * AH 53, Nr. 70 Fer. 2 213r Sancti spiritus adsit * AH 53, Nr. 70 Fer. 3 214r Sancti spiritus adsit … AH 53, Nr. 70 Dnca pentecostes et de spiritu sancto 69rb 1v 39r 39v Summi triumphum regis … AH 53, Nr. 67 In ascensione dni 64rb 1v 36r 36v Veni sancte spiritus … AH 54, Nr. 153 Dnca pentecostes et de spiritu sancto 210r Victimae paschali laudes * AH 54, Nr. 7 Dnca 2. post pascha 192v Victimae paschali laudes * AH 54, Nr. 7 Dnca 3. post pascha 194r Victimae paschali laudes … AH 54, Nr. 7 Dnca resurrectionis dni Victimae paschali laudes … AH 54, Nr. 7 Dnca in albis = Dnca 1. post pascha 32v 207r 25v 186r 2. Sanctorale Incipit Nachweis Anlass Cod. 100 GN II * nur Verweis, ** Inc. mut. Ad laudes salvatoris * AH54, Nr. 88 Silvester papa 95r Agni * AH 53, Nr. 50 Georgius 123v Agni * AH 53, Nr. 50 Monica 128v Agni paschali esu … AH 53, Nr. 50 Ambrosius 120r Agni paschalis * AH 53, Nr. 50 Pancratius et socii 132v Agni paschalis * AH 53, Nr. 50 In translatione s. Albuini 132v Agni paschalis * AH 53, Nr. 50 Victor 132v Agone * AH 53, Nr. 229 Achatius et socii 133v Agone triumphali * AH 53, Nr. 229 Vitus, Modestus et Crescentia 133r Agone triumphali * AH 53, Nr. 229 Tiburtius 171v Caeli enarrant gloriam Dei … AH 50, Nr. 267 In divisione apostolorum 151v Caeli regem attollamus … AH 55, Nr. 62 Anna 160r Nr. 139 Nr. 940 408 Incipit Verzeichnis der Sequenzen Nachweis Anlass Cod. 100 GN II Nr. 139 Nr. 940 * nur Verweis, ** Inc. mut. Clare sanctorum * AH 53, Nr. 228 Thomas ap. 85r Clare sanctorum * AH 53, Nr. 228 Barnabas ap. 133r Clare sanctorum * AH 53, Nr. 228 In commemoratione s. Pauli 141v Clare sanctorum * AH 53, Nr. 228 In octava ap. s. Petri et Pauli 145r Clare sanctorum * AH 53, Nr. 228 Iacobus 159v Clare sanctorum * AH 53, Nr. 228 Petrus ad vincula 165r Concentu parili … AH 53, Nr. 99 In purificatione BMV 104r Congaudentes exultemus vocali … AH 54, Nr. 66 In translatione s. Nicolai 145r Deus in tua virtute … AH 53, Nr. 122 Andreas 77v Dixit dominus ex Basan … AH 50, Nr. 269 In conversione s. Pauli 100r Exultent filiae Sion * AH 50, Nr. 271 Barbara 79r Exultent filiae Sion * AH 50, Nr. 271 Dorothea 110r Grates Deo et honor … AH 53, Nr. 119 Gratuletur gloriose unitas … Afra et sodales 165v Barbara 79v Haec est sancta sollemnitas … AH 53, Nr. 56 Inventio s. crucis 126v Hanc concordi famulatu … AH 53, Nr. 215 Stephanus 87r AH 9, Nr. 236 Ingenuinus et Albuinus episcopi 107v Innoventur hac in die … 82v 100v 97r 15v 7v Iohannes Iesu Christo … AH 53, Nr. 168 Iohannes ev. 90v 18v 10v Laude Christo debita … AH 55, Nr. 265 Nicolaus 81r 102r 98v 89r 86v Laudes crucis * AH 54, Nr. 120 Inventio s. crucis 126v Laudes salvatori * AH 53, Nr. 36 Philippus et Iacobus ap. 124v Laudes salvatori voce … AH 53, Nr. 36 Ambrosius 117r Laurenti David … AH 53, Nr. 173 Laurentius Laus sit regi gloriae … AH 55, Nr. 51 Agnes Laus tibi Christe patris … AH 53, Nr. 157 Innocentes 93v 21r 13r Laus tibi Christe qui es … AH 50, Nr. 268 Maria Magdalena 156r 86r 83v Mundi decor mundi forma … AH 55, Nr. 244 Martha 162r Omnes sancti Seraphin Cherubin throni … AH 53, Nr. 112 Omnes santi Petre summe Christi … AH 53, Nr. 210 Petrus et Paulus ap. 140r 83v 80v Regi psalmistae citharistae … AH 54, Nr. 64 Margaretha 148v Sancti Baptistae Christi … AH 53, Nr. 163 Iohannes Baptista 136r 81r 78r Spe mercedis * AH 55, Nr. 9 Thomas mart. 95r Summi triumphum * AH 53, Nr. 67 In translatione s. Albuini 132v Summi triumphum * AH 53, Nr. 67 Victor 132v Summi triumphum regis … (Ascensio) AH 53, Nr. 67 Ambrosius 121v 169v 74vb 98r** 87rb 409 Verzeichnis der Sequenzen Incipit Nachweis Anlass Cod. 100 GN II 78ra 142r Nr. 139 Nr. 940 * nur Verweis, ** Inc. mut. Veni praecelsa domina … AH 54, Nr. 193 In visitatione BMV Verbum Dei Deo natum … AH 55, Nr. 188 Iohannes ante Portam Latinam Victime paschali * AH 54, Nr. 7 Philippus et Iacobus ap. Victime paschali * AH 54, Nr. 7 128v 107r Inventio s. crucis radiert, nicht mehr leserlich Marcus ev. 108r 33r GN I GN II 123v 3. Commune Sanctorum Incipit Nachweis Anlass Cod. 100 Cod. 154 Nr. 139 Nr. 940 143v 133r * nur Verweis, ** Inc. mut. Ad laudes salvatoris … AH 54, Nr. 88 De simplici confessore / De confessore et pontifice / De uno confessore Agone triumphali … AH 53, Nr. 229 De (pluribus) martyribus 136v Clare sanctorum senatus … AH 53, Nr. 228 De apostolis 131r Clare sanctorum senatus … AH 53, Nr. 228 De uno apostolo 7r Dilectus Deo et hominibus … AH 54, Nr. 90 De uno confessore 56bisv Exultent filiae Sion … AH 50, Nr. 271 De virginibus Hic oculis ac manibus … AH 9, Nr. 388 De uno confessore Hic oculis ac manibus* AH 53, Nr. 181, Str. 15 (Divisio) De simplici confessore aut evangelista O beata beatorum … AH 55, Nr. 14 De (pluribus) martyribus Omnes sancti seraphin cherubin … AH 53, Nr. 112 In dedicatione unius altaris Plausu chorus laetabundo … AH 55, Nr. 6 De uno apostolo Psallat ecclesia mater illibata … AH 53, Nr. 247 In dedicatione ecclesiae Rex regum Deus noster … AH 53, Nr. 243 De simplici confessore 145v Spe mercedis et coronae … AH 55, Nr. 9 De uno martyre 140r 129v AH 54, Nr. 7 De uno beato in tempore paschali / De uno martyre in tempore paschali 106v 31r Victime paschali * 2v 149v 57r 27v 120r 138v 71r 59v 136r 135r 124v 29r 267r** 8r 85va 3r 50v 50v 262v 46v 410 Verzeichnis der Sequenzen 4. De BMV Incipit Nachweis Cod. 100 Anlass Cod. 154 Sign. 15063 Nr. 139 Nr. 940 GN II 70r 66v 104r 75r 71r 172r * nur Verweis Ave Maria gratia plena dominus tecum … AH 40, Nr. 119 De BMV Concentu parili … AH 53, Nr. 99 In purificatione BMV 10va Conceptio Mariae virginis … AH 54, Nr. 188 In conceptione BMV 73va Congaudent angelorum chori … AH 53, Nr. 104 In assumptione BMV 79vb 3v Divisio von AH 53, Gaude Dei genitrix * Nr. 15 Infra nativitatem et purificationem 67v Hodiernae lux diei … AH 54, Nr. 219 In visitatione BMV 79v Hodiernae lux diei celebris … AH 54, Nr. 219 De BMV 3v 76r Laetabundus exultet AH 54, Nr. 2 fidelis … Infra nativitatem et purificationem. 68r 63v Mittit ad virginem … AH 54, Nr. 191 In adventu domini aut in annuntiatione angelica 65v 61r Salve mater salvatoris * AH 54, Nr. 245 De BMV Stabat mater dolorosa iuxta crucem lacrimosa … AH 54, Nr. 201 De compassione BMV Stirpe Maria regia … AH 53, Nr. 95 In nativitate BMV 83ra 77r 73v 186v Veni praecelsa domina … AH 54, Nr. 193 In visitatione BMV 78ra 78r 75r 142r Verbum bonum et suave … AH 54, Nr. 218 De veneracione s. Marie per annum 57r 64v 58v Virgini Mariae laudes … AH 54, Nr. 18 In annuntiatione BMV circa festa pascalia 73v Virgini Mariae laudes … AH 54, Nr. 18 Post pasca de s. Maria officium 77v 193v 70r 411 Verzeichnis und Abbildungen der Abreibungen Vorbemerkung: Alle Abbildungen einzelner Stempel und Rollen in Originalgröße, Ausnahme bildet eine Stempelanordnung im Mittelfeld des Hinterdeckels vom ersten Band des Graduale Neocellense I (Tafel VII). Aufgrund der Größe musste die Abreibung auf 75% verkleinert werden. Wiedergegeben werden alle Stempel, Rollen und Platten der verzierten Einbände. Fotos der Einbände sind im Handschriftenportal www.manuscripta.at veröffentlicht. Die Anordnung erfolgt (mit wenigen Ausnahmen) nach den Signaturen der Handschriften und nicht nach Werkstätten oder Motiven; die angeführten Nummern beziehen sich auf die Beschreibungen im Katalog. Taf. I, Abb. 1: Cod. 123: Nr. 1: kreisförmiger Stempel Taf. I, Abb. 2: Cod. 142: Nr. 1: Rolle Ranke mit eingeschlossenen Blüten und Granatapfel; Nr. 2: Rolle Ranke mit eingeschlossenen Blüten und Granatapfel; Nr. 3: Rolle Rundbogen auf Stab; Nr. 4: zwölfblättrige Rosette mit zwölfblättriger Mittelblüte frei; Nr. 5: florales Dekor frei Taf. I, Abb. 3: Cod. 194: Nr. 1: Rolle verschränkte Bögen mit floralem Dekor; Nr. 2: Rolle Sichel mit eingeschriebenem Blattmotiv abwechselnd mit floralem Dekor; Nr. 3: Rolle mit Kornblumenmotiv Taf. I, Abb. 4: Cod. 204: Nr. 1: Drache (Vogel?) in Raute; Nr. 2: fünfblättrige Blüte frei; Nr. 3: Schriftband „Maria“; Nr. 4: Einhorn in Raute; Nr. 5: doppelköpfiger Adler in Raute Taf. II, Abb. 5: Cod. 353 und 654: Nr. 1: Rolle Enthauptung des Johannes, Eherne Schlange, Kreuzigung, Auferstehung; Nr. 2: Platte Erzengel (Michael?) mit Wappenschild von Propst Augustin Schabl (Propst 1569–1581) mit zwei steigenden geflügelten Löwen, darüber Inschrift Augustinus praepositus Novæcelle; Nr. 3: florales Dekor aus Vase wachsend; Nr. 4: Kreuzigung Christi; Nr. 5: florale Rolle Taf. II, Abb. 6: Cod. 405: Nr. 1: Rolle mit in stilisierten Blättern auslaufender Volute; Nr. 2: rautenförmiger floraler Einzelstempel; Nr. 3: gebündeltes Dreiblatt in Quadratform; Nr. 4: fünfblättrige Blüte frei; Nr. 5: florales Dekor in Rhombusform; Nr. 6: achtblättrige Blüte frei Taf. II, Abb. 7: Cod. 440: Nr. 1: kleine Siebenpunktblüte; Nr. 2: Schriftband „Maria“; Nr. 3: Doppeladler mit Krone in Tropfenform; Nr. 4: Rosette mit zweifachem Blütenkranz frei; Nr. 5: Kielbogen mit Blüte frei; Nr. 6: nach links blickender Steinbock in Rhombus Taf. III, Abb. 8: Cod. 397: ganzer Einband (Vorderdeckel und Hinterdeckel) 412 Verzeichnis und Abbildungen der Abreibungen Taf. IV, Abb. 9: Cod. 726: ganzer Einband (Vorderdeckel = Hinterdeckel) Taf. IV, Abb. 10: Sign. 15063: Nr. 1: Rolle Profilköpfe in Handspiegeln mit Quastenbehang; Nr. 2: Rolle Eherne Schlange, Kreuzigung, Auferstehung; Nr. 3: Rolle Menschenpaare unter Baldachin mit floralem Dekor; am Ende der Rolle jeweils Umschrift DVRKO; Nr. 4: Herzblatt; Nr. 5: Doppellilie mit rhombischem Mittelblatt; Nr. 6: Rolle florales Dekor; Nr. 7: Rolle Ranke mit eingeschlossener Fünfpunktblüte Taf. V, Abb. 11: Nr. 139 (olim Sign. 14819): Nr. 1: vierblättrige Blüte frei; Nr. 2: Adler in Rhombus; Nr. 3: Einzelstempel (stark abgerieben, nicht mehr erkennbar) in Kreis; Nr. 4: vierblättrige Blüte; Nr. 5: Lilie in konkavem Karo; Nr. 6: sechsblättrige Blüte in Kreis; Nr. 7: Lilie frei; Nr. 8: fünfblättrige Blüte frei; Nr. 9: Dreiblatt frei Taf. V, Abb. 12: Nr. 940: Nr. 1: vierblättrige Blüte frei; Nr. 2: gebogener Ast; Nr. 3: punktiertes Herz frei; Nr. 4: sechsblättrige Blüte in Kreis; Nr. 5: rautenförmige Lilie frei; Nr. 6: Punktdekor in Karo; Nr. 7: Astdekor in Rechteck; Nr. 8: Osterlamm mit Fahne in Kreis; Nr. 9: Vierblatt in konkavem Karo Taf. V–VII, Abb. 13: Graduale Neocellense I: Nr. 1: Rolle einseitwändig verschränkte Bögen mit Lilienbekrönung, darüber Leiste aus fünfblättrigen Blumen; Nr. 2: Rolle Profilköpfe in Kreis im Wechsel mit Schriftband und begleitendem floralem Dekor; Nr. 3: Rolle Wappen von Neustift (Tau-Kreuz), Säben und Patriarchenkreuz einander abwechselnd, verbunden mit Rankendekor in gegenständigen Dreipassformen; Nr. 4: Rolle Profilköpfe in Handspiegeln mit Quastenbehang; Nr. 5: eng aneinander gereihte Einzelstempel Schriftband „Maria“, dazwischen Fischblasenmotiv und in den Zwickeln Trifolien; Nr. 6: Maske frei; Nr. 7: Lilie frei; Nr. 8: Stempelblüte mit Rautenranke; Nr. 9: Rolle ineinander greifende, aus drei Linien aufgebaute Bögen mit Lilienbekrönung und zwei eingeschlossenen vierblättrigen Blüten; Nr. 10: Lilie in Raute; Nr. 11: Brokatmotiv (mittig zum Vierblatt angeordnet, oben und unten jeweils ein Brokatmotiv geschlossen); Nr. 12: florales Dekor, rautenförmig; Nr. 13: florales Dekor mit geschlossener Knospe in Rautenform; Nr. 14: Brokatmotive offen TAFEL I 413 Cod. 142 Cod. 123 Nr. 1 Nr. 4 Nr. 3 Nr. 5 Nr. 1 Cod. 194 Nr. 2 Cod. 204 Nr. 1 Nr. 2 Nr. 3 Nr. 1 Nr. 2 Nr. 3 Nr. 4 Nr. 5 TAFEL II 414 Cod. 353 und 654 Nr. 3 Nr. 4 Nr. 1 Nr. 2 Nr. 5 Cod. 405 Cod. 440 Nr. 4 Nr. 1 Nr. 4 Nr. 2 Nr. 2 Nr. 5 Nr. 5 Nr. 1 Nr. 3 Nr. 6 Nr. 3 Nr. 6 TAFEL III Cod. 397 Vorderdeckel Hinterdeckel 415 TAFEL IV 416 Cod. 726 Sign. 15063 Vorderdeckel = Hinterdeckel Nr. 4 Nr. 1 Nr. 2 Nr. 5 Nr. 7 Nr. 6 Nr. 3 TAFEL V 417 Cod. 139 (olim Sign. 14819) Nr. 1 Nr. 2 Nr. 3 Nr. 6 Nr. 5 Nr. 4 Nr. 7 Nr. 8 Nr. 9 Nr. 940 Nr. 6 Nr. 1–3 Nr. 4 Nr. 5 Nr. 7 Nr. 8 Nr. 9 Graduale Neocellense I Nr. 10 Nr. 13 Nr. 14 Nr. 12 TAFEL VI 418 Nr. 6 und 7 Nr. 3 Nr. 2 Nr. 5 Nr. 4 Nr. 9 Nr. 1 Nr. 8 und 7 Nr. 8 TAFEL VII 419 Nr. 11 (75 %) REGISTER 423 TEXTANFÄNGE (INCIPIT) A cunctis iniquitatibus nostris exue nos domine et intercedente beata et gloriosa Dei genitrice virgine Maria 654, 10r A cunctis nos quaesumus domine mentis et corporis defende periculis et intercedente gloriosa virgine Dei genitrice 405, 220r A domo tua quaesumus domine spiritales nequitiae repellantur et aeriarum discedat maligni 654, 95v A haec dictio. A Interdum includit ut C de 4, 1va Ab infirmitate surgentibus cibaria nociva petentibus non est denegandum 748, Iv Abicit mundus pauperes et honorat divites. Nota Augustinus dicit super quod corvus est illius naturae quod pullos suos 52, 202ra Abigeus dicitur ille qui pecora alicuius a pascuis et armentis subducit … Abolitio est ab accusando cessatio 155, 18ra Abiit Iesus trans mare Galilaeae quod est Tiberiadis et sequebatur eum multitudo magna etc. (Io 6,1). In hoc evangelio considerantur triplices personae scilicet Christus discipuli et turbae 53, 242vb – Licet secundum Augustinum in quodam sermone ad fratres Non debeamus loqui sublimia 53, 242rb Abrahae sinus dicitur requies sanctorum qui olim fuit limbus nunc est caelum et sic orat ecclesia sinus te suscipiat 617, 196r Absolve quaesumus domine animam fratrum et sororum parentum et benefactorum nostrorum et omnium fidelium defunctorum 405, 128v Absolve domine quaesumus animas famulorum famularumque tuarum ab omni vinculo delictorum 405, 206v Absque synagogis facient vos (Io 16,2). Sciendum quod sicut Iudaei olim tenuerunt se ad discipulos Christi scilicet separando eos a participatione et communione ipsorum 153, 131vb Absterget Deus omnem lacrimam etc. (Apc 7,17 und 21,4). Ad imitationem sanctorum flere debemus et nos quinque de causis 78, 97vb Abstinentia est meriti augmentativa. Sapientiae acquisitiva 125, 9ra Accepta sit quaesumus domine in conspectu tuo nostra devotio ut passionum charismatum et gloriae rationalis ac beatissimae animae 353, 192r Accidens morbi est triplex scilicet praeventivus ut rigor concomitans ut dolor capitis 748, 40v Accipe itaque quascumque exiles quascumque meritis tuis impares gratiarum actiones 405, 209v Accipe o homo exemplum qui es in periculis huius mundi a sancto Andrea 617, 195r Accipite et manducate ex hoc omnes (1 Cor 11,24). Bene fit illi homini qui hunc panem digne et recte manducat 617, 173r – Nota quando tritulator est debilis et habet grave flagellum in manu 174, 187rb Accipite spiritum sanctum quorum remiseritis peccata Ioh. 20 (22). Salvator noster Christus dominus sicut omnia in sua passione ostendit ut declararet suam humanitatem 53, 318rb Actiones nostras quaesumus domine aspirando praeveni et adiuvando prosequere 252, 24r; 405, 128r, 225r Ad XVI numerum quintum dato primum 360, 26v Ad cognoscendam naturam constructionis in se et in suis partibus sive speciebus. Sciendum quod constructio est quoddam incomplexum et quia incomplexum secundum Boetium cognoscitur dupliciter. Definitione quae ordinatur ad cognoscendam rem in se 605, 5va Ad eum veniemus et mansionem apud eum faciemus (Io 14,23). Sciendum quod hodierna festivitas inter alia festa dignitatem singularem obtinet 153, 134va Ad gaudia supernorum perducat nos regina caelorum 405, 210r Ad imitationem sanctorum flere debemus et nos quinque de causis 78, 97vb Ad mensam magnam sedisti (Sir 31,12). Scito quod talia oportet te praeparare qualia sunt in hostia. Quanto maior est festivitas tanto se melius homines praeparare debent 177, 75rb Adest tibi venerabile tempus Quadragesimae 617, 143r 424 Adesto Deus unus omnipotens – Anthrax et carbunculus Adesto Deus unus omnipotens pater et filius et spiritus sanctus te unum in substantia trinum 353, 117v Adesto domine supplicationibus nostris ne sit a nobis famulis tuis longinqua tua miseratio 654, 28v Adesto nobis domine Deus noster et quos sanctae crucis laetari fecisti 405, 221v Adesto supplicationibus nostris omnipotens Deus et quibus fiduciam sperandae pietatis 353, 49v Adesto supplicationibus nostris quaesumus omnipotens Deus et quibus fiduciam sperandae pietatis indulges 405, 217v, 218v Adiutorium nostrum in nomine domini 405, 129r Adiuvent nos quaesumus domine et haec mysteria 154, 4r Adiuvet nos quaesumus domine beatae Mariae semper virginis intercessio veneranda 405, 211r Aetatem habet ipse de se loquatur (Io 9,21). Pro huius thematis introductione tres scholas invenio scilicet examinationis, confessionis, obligationis. Prima schola est examinationis et in ista schola legitur quod reus dato quod omnia sunt allegata et approbata 597, 34ra Affer manum tuam et mitte in latus meum (Io 20,27). Benigni homines habent modum dum vident aliquem carum amicum cecidisse in aliquam passionem solent eos instruere 617, 136r – Benigni homines habent modum dum vident aliquem ex caris amicis suis cecidisse in aliquam passionem solent eos instruere 78, 8vb Ait vere dominus est in loco isto et ego nesciebam (Gn 28,16). His verbis possumus quattuor notare. Primo quis sit Iacob secundo quod dormivit tertio quae sit scala quarto quod angelos vidit 162, 3va Aliquis cognovit matrem et filiam vel duas sorores vel duas consanguineas mulieres 155, 8va Aliquis confitetur aliquod crimen sacerdoti et episcopus excommunicat omnes huius criminis consocios 155, 9rb Aliquis confitetur peccata sua sacerdoti et dicit quod non potest cessare 155, 9rb Aliquis contraxit sponsalia cum aliqua muliere et contracto matrimonio cognovit carnaliter mulierem sponsae ante sponsam 155, 8vb Aliquis dicit confessori suo quod mulierem quam nunc habet duxit in uxorem in facie ecclesiae 155, 8vb Aliquis est surdus et mutus vel ita aeger quod non potest peccata sua confiteri sacerdoti 155, 9rb Aliquis fuit testis in causa matrimoniali qui falsum protulit testimonium vel aliqua dixit de causa matrimoniali se scire quae in veritate ignoravit 155, 8rb Aliquis furiosus interfecit hominem postea factus sanae mentis se hominem interfecisse confitetur 155, 9ra Aliquis habens uxorem postmodum commisit cum filia uxoris suae adulterium 155, 8va Aliquis habetur pro praelato et in veritate non est praelatus audit confessiones parrochianorum illius parrochiae 155, 9va Aliquis in infirmitate petit sacerdotem et antequam ad eum venerit sacerdos 155, 9va Aliquis peregrinus vel viator seu negotiator vel mercator venit ad extraneum sacerdotem 155, 9rb Aliquis segetem meam sevit tempore opportuno animo surripiendi sed nihil de ea asportavit 155, 9ra Aliquis venit ad confessorem et dicit ei peccata sua et dicit se non posse abstinere a luxuria 155, 9ra Aliquis vovebat perpetuam continentiam postmodum non obstante voto 155, 8va Alliaseph puncti rubei qui sunt ut pulicis morsus et fiunt in omnibus membris 748, 213ra Alterius circumventio alteri non praebet actionem. Actus legitimi ut supra. Aliud est vendere aliud venditioni consentire 614, 275r Ambrosius cum incunabulis in atrio praetorii esset positus et dormiret 308, 22vb Amen Amen dico vobis. Si quid petieritis patrem in nomine meo dabit vobis (Io 16,23). Quamvis multis modis homo serviat Deo et pro peccatis suis satisfaciat 53, 338rb An maritus uxori adulterae subtrahere possit debitum cohabitationum et dotem sine iudicio ecclesiae. Tua fraternitas hic tractantur iura an vir uxori adulterae 155, 6vb An reddituarii teneantur propriis sumptibus deferre dominis redditus et praesertim ecclesiarum. Et videtur quod sic quia redditus ecclesiae videntur esse dos ipsius 155, 7va Animabus quaesumus domine omnium fidelium defunctorum oratio proficiat 654, 66r Animam meam pono pro ovibus meis (Io 10,15). Hic dicendum est de caritate Dei quam habet ad nos et etiam de caritate quam nos ad ipsum habere debemus 153, 109vb Animas pauperum tuorum ne obliviscaris in finem (Ps 74,19). Notandum quod quintuplex est anima scilicet anima immunda anima rapta anima sepulta anima salvata anima cruciata 617, 190r Anno salutis MDCLIII septimo Kalendis Septembris ad maiorem Dei gloriam S. P. N. Augustini honorem 76, 74r Annuntio vobis gaudium magnum … (Lc 2,10). Scire debetis in Christo dilectissimi quod dies hodierna ex salvatoris nostri nativitate nobis redditur gaudiosa 153, 4va Ante diem festum paschae sciens Iesus quia venit hora eius etc. (Io 13,1). Chrysostomus super Iohannem tractans illud Ioh. 10. Facta sunt encaenia 53, 265ra – Sicut auris non potest satiari auditu nec oculus 53, 265va Anthrax est apostema quod partim fit de cholera partim de sanguine partim de melancholia. De sanguine habet putredinem 748, 90r Anthrax et carbunculus quia fiunt ex materia grossa 748, 54rb Anticipatio paroxysmi augmentum – Beati Petri apostoli 425 Anticipatio paroxysmi augmentum non significat nisi cum hoc longitudo temporis aut fortitudo febris augeatur 748, 1r Apostolicis nos quaesumus domine beatorum apostolorum Petri et Pauli et aliorum apostolorum attolle praesidiis 405, 217r Arescentibus hominibus prae timore et exspectatione quae supervenient universo orbi (Lc 21,26). In praecedenti sermone dixi de signis adventus diei iudicii 53, 13ra Arguet mundum de iustitia (Io 16,8). Glossa semper quam non habet. Unde hic est dicendum quomodo homo debet iustus inveniri si vult salvari 153, 119vb Arguet mundum de peccato quia non credunt in me (Io 16,8). Glossa semper quod habet. Unde hic est dicendum quomodo quis committit septem peccata mortalia 153, 119vb – Sciendum quod plura genera hominum sunt qui dicunt verbis se credere in Deum 153, 115va – Sciendum quod sicut Deus arguet peccatores de suis excessibus in futuro 153, 117vb Aristoteles 2o De anima Simile a simili nutritur sicut patet in cibo digesto a quo corpus nutritur 153, 81rb Aristoteles 1o Ethicorum finis per quem omnia desiderantur et sperantur optimus est 153, 7rb Aristoteles 8o Ethicorum dicit Vera amicitia habet reamationem. Et quia Christus vere nos dilexit 153, 77ra Aristoteles 9o Metaphysicae Contraria non fiunt in eodem. Cum ergo spiritus sanctus intrat in cor hominis tunc mala et peccata depellit 153, 137ra Aristoteles 5o Physicorum Omnis motus tendit ad aliquem terminum per debita media 153, 74va Aristoteles 8o Physicorum Illud est prius quo non existente non sunt alia sed sine fide 153, 67va Aristoteles In sempiternis nihil violentum. Qui ergo alte in caelo vult 153, 46vb Ars praedicandi est scientia docens de aliquovis termino vel ente aliquid dicere 155, 139r Artat statutum doctores legentes et ut quaestiones disputatas 614, 284r Ascendente Iesu in naviculam etc. (Mt 8,23). Christus dominus tempore quo suam praedicationem incepit semper operatus est 53, 115ra – Licet mundus iste primo appareat valde amicus tranquillus et pacificus hominibus 53, 114va Asma ut dicit Avicenna est aegritudo pulmonis cum qua patiens non invenit excusationem ab anhelitu … Et Rasis addit tussim et quod laeditur ex iacendo 123, 212r Assumpsit Iesus duodecim discipulos suos et ait illis Ecce ascendimus Hierosolymam et consummabuntur omnia etc. (Lc 18,31). Caelestis medicus quamvis vocatus ab infirmo homine propter originale peccatum 53, 145vb – Hoc evangelium facit memoriam de Christi passione et secundum Wilhelmum Duranti 53, 152ra – In praesenti evangelio ponuntur duo principalia. Primum de Christi passione 53, 146rb – Secundum scripturas sanctorum homo per peccatum fit instabilis et mutabilis et despectus 53, 151va Audi Israel Deus tuus unus est (Dt 6,4). Sunt autem septem genera hominum qui Deum non colunt sed idola adorant. Primus genus est de quibus 308, 210ra Audite haec omnes gentes auribus percipite omnes qui habitatis orbem 19, 1ra Audite haec quae dicturus sum. Hic attentos facit omnes Gregorius id est omnes iniqui, haec iam hodie 19, 1ra Auge in nobis domine fidem rectam spem firmam caritatemque perfectam 405, 224r Augustinus dicit in De consensu evangelistarum et magistri historiarum quod Herode mortuo 53, 76va Augustinus in quodam sermone de Christi nativitate qui incipit Adiuvet nos humillimos 53, 57va Augustinus Non solum Christi verba sunt nobis in documentum sed etiam Christi facta sunt nobis in exemplum 153, 30va Aures tuae pietatis mitissime Deus inclina precibus meis 154, 6r; 405, 223v Aut fratres sunt ex utroque coniuncti aut non. Si ex utroque coniuncti et tunc aut soli fratres exstant 155, 106r Ave gloriosa Maria mater Christi virgo pia pone misericordiam et compassionem 353, 161v Ave gratia plena dominus tecum (Lc 1,28) benedicta tu in mulieribus (Lc 1,42). Dixit angelus ad virginem de caelo missus. Et benedictus fructus ventris tui. Adicit Elisabeth spiritu prophetico inspirata 31, 49ra Ave gratia plena dominus tecum (Lc 1,28). Ista dies praecipue maior est omnibus festivitatibus 174, 155ra – Si quis princeps vel rex vel magnus dominus vult destinare nuntium vel legatum ad summum principem vel regem vel dominum magnum pro magna et ardua causa 31, 67rb Avete omnes fideles animae in sancta pace quiescite qui vos redemit suo pretioso sanguine vobis dignetur misereri poenasque vestras 654, 91r Avete omnium fidelium animae in pace Christi requiescite quos redemit suo pretioso sanguine 405, 220r Beata virgo comparatur caelo. Caelum enim dicitur quasi gaza heliorum id est solis domus, quia quasi in domo propria 171, 62ra Beatae et gloriosae semperque virginis Dei genitricis Mariae quaesumus omnipotens Deus intercessio gloriosa nos protegat 405, 216v Beati Petri apostoli tui quaesumus domine intercessione nos protege et animas famulorum tuorum sacerdotum sanctorum tuorum iunge 405, 128v, 129r 426 Beati qui habitant – Chrysostomus super Iohannem Beati qui habitant in domo tua domine (Ps 84,5). David dicit haec verba. Notandum est quod domus domini est quadruplex scilicet domus materialis 162, 2rb Beatus Bernardus in sermone De quadragesima dicit quod septem sunt impedimenta 53, 134ra Beatus Thomas sanctitatem suam ita subtiliter occultabat ut salvo semper honestatis rigore 308, 1vb Beatus vir qui non abiit / Pestilentiae (darüber:) Der Suechte 308, 203ra Beatus vir. Pestilentiae contagie (!) quod dum unum apprehendit celeriter transit ad plures quia velut incendium 155, 141r Bene fit illi homini qui hunc panem digne et recte manducat 617, 173r Benedicta tu in mulieribus (Lc 1,42). Dixit angelus ad virginem de caelo missus. Et benedictus fructus ventris tui. Adicit Elisabeth spiritu prophetico inspirata 31, 49ra – Illud digne commendatur quod in se consideratum bonum apparet et aliis comparatum excellit 31, 84va – Si volumus amplius scire quod multipliciter est sanctissima virgo domina Maria benedicta inspiciamus 31, 89rb Benedictionem omnium gentium dedit illi (Sir 44,25). In verbis istis ostendit nobis spiritus sanctus quaedam signa in beato Nicolao propter quae signa obtinuit benedictionem omnium gentium 78, 4rb Benedictum sit dulce nomen domini mei Iesu Christi et gloriosissimae virginis Mariae matris eius in saeculum 617, 195r Benedictus qui venit in nomine domini (Mt 21,9; Mc 11,9; Lc 13,35; Io 12,13). Hodie sancta mater ecclesia duas processiones recolit 153, 79rb Beneficium ecclesiasticum non potest licite sine institutione canonica obtineri 614, 282v Benigni homines habent modum dum vident aliquem carum amicum cecidisse in aliquam passionem solent eos instruere 617, 136r Benigni homines habent modum dum vident aliquem ex caris amicis suis cecidisse in aliquam passionem solent eos instruere 78, 8vb Benignitas salvatoris Dei nostri maior est in exaudiendo petitiones filiorum suorum 153, 119vb Bonorum honorabilium notitiam opinantes magis autem alteram altera aut secundum certitudinem aut ex eo 163, 32r Caelesti quaesumus domine lumine semper et ubique nos praeveni rationalis 353, 187r Caelestis medicus quamvis vocatus ab infirmo homine propter originale peccatum 53, 145vb Carissime frater. Vivatis in religione secundum doctrinam apostoli dicentis Sobrie et iuste et pie vivamus in hoc saeculo exspectans beatam spem et adventum gloriae magni Dei. In quibus verbis apostolus Paulus docet 617, 55r Caro mea vere est cibus … (Io 6,55). Sic legimus quod unigenitus Dei filius volens [ 153, 156vb – Sicut dicit Aristoteles 4o Ethicorum Terribilissimum omnium terribilium videtur esse mors 153, 154ra Cassiodorus in libro De anima. Magistri scholarum litterarum aiunt animam esse substantiam simplicem speciem naturalem distantem a materia corporis sui organum membrorum 597, 57vb Casus talis est. Quaedam domus religiosae quarum fratres non manducant emerunt pensiones annuas perpetuas ipsis opportunas 447, 208ra Ceperunt ramos palmarum et processerunt obviam ei (Io 12,13). Pro huius thematis introductione invenio tres scilicet olivae in Minerva, lauri in Mercurio, palmae in Marte. Prima schola est olivae in Minerva. Nam ut possum capere ex dictis poetarum oliva consecrata est in nomine Minervae 597, 40vb Christe rex clementissime fili Mariae qui inter plurima opprobria sanctissimum spiritum emittendo patri tuo 353, 114r Christi nomen invocans ad honorem ipsius et reverentissimi patris domini Bononiensis archidiaconi qui divinam potentiam imitatus de me nihil fecit aliquid 99, 32va Christus dominus frequenter in suis sermonibus arguit Iudaeis super peccatis suis 53, 246va Christus dominus ne dum aetate virili sed etiam in pueritia sua se nobis ita exhibet ut ostendat quanta caritate nos diligat 53, 76ra Christus dominus per scripturas nobis valde incutit timorem de extremo iudicio 53, 6va Christus dominus tempore quo suam praedicationem incepit semper operatus est 53, 115ra Christus dominus volens omnem dubitationem de sua divinitate a nobis excludere 53, 96va Christus ieiunavit nobis in doctrinam et exemplum quia secundum Gregorium Omnis Christi actio nostra est instructio 153, 62va Christus nobis tribuit normam (!) hic vivendi / Et exemplo docuit viam gradiendi / Ut mundana vanitas possit vilipendi 165, 143va Chrysostomus et Theophilus dicunt et habetur Matthaei 12 Ille daemoniacus erat mutus 53, 240va Chrysostomus homilia 16 super Matthaeum dicit Omnium malorum tria et fortiora sunt tria mala 53, 185va Chrysostomus super Iohannem tractans illud Ioh. 10. Facta sunt encaenia 53, 265ra Chrysostomus super Matthaeum – Corda nostra quaesumus 427 Chrysostomus super Matthaeum homilia 28 Homo Christus paterfamilias cui caeli et pater (recte: terra) una est domus familia 53, 124va Circa distinctionem sacramenti (!) quaeritur utrum haec distinctio sit bona. Sacramentum est invisibilis gratiae visibilis forma ut imaginem gerat et causa existat. Respondeo quod sic quia completissime rationem sacramenti designat 77, 1ra Circa initium huius libri sicut aliorum sunt aliqua generalia autem textus in titulationem praenotanda. Primo in quali scientia 163, 32r Circa nativitatem domini tria notantur. Primo natus describitur secundo nativitatis tempus tertio nativitatis utilitas. De primo sciendum quod fuit natus admirabilis ineffabilis formidabilis venerabilis 617, 136r Circa quartum librum Sententiarum primo quaeritur quid sit sacramentum. Dicendum quod sacramentum quattuor modis solet describi vel definiri. Primo sic sacramentum est sacrae rei signum 52, 112r Circa quartum Meteorum. Utrum tamen quattuor sunt qualitates primae scilicet caliditas frigiditas siccitas et humiditas. Et arguitur primo quod non quia 440, 411va Circa quem textum est dicendum an sancti in aeterna beatitudine quam habebunt post resurrectionem sedebunt vel stabunt 53, 346vb Circa quod sciendum quod sunt sex quae debent hominem retrahere ab omni peccato mortali 153, 96ra Circa textum notandum quod philosophus in textu dicit quod erit determinandum 163, 47r Circumcisionis dies est sollemnis quia octavus nativitatis Christi est. Voluit autem dominus die octava circumcidi unde Rabbi Moyses philosophus et theologus licet Iudaeus 617, 138v Coitus maxime exsiccat quia per ipsum recedit magna humiditas 155, 138r Collegerunt pontifices et Pharisaei concilium adversus Iesum (Io 11,47). Pro huius thematis introductione invenio tres scholas scilicet urbis fundatorum, in campo bellatorum, in campo clericorum. Prima schola est urbis fundatorum 597, 38rb Commendo domine in misericordiam tuae divinitatis maiestatis ac magnificentiae 440, urspr. VDS (heute abgelöst) Commonefacio vos enim fratres mei collegaeque devoti futuri festi nativitatis Christi ad sacras ordines 756, 53rb Concede misericors Deus fragilitati nostrae praesidium ut qui sanctae Dei genitricis Mariae 353, 149r; 405, 215v Concede nos famulos tuos quaesumus domine Deus noster perpetua mentis et corporis sanitate gaudere 353, 143r; 405, 212v, 218r Concede quaesumus omnipotens Deus sanctum nos spiritum vocis promereri 353, 29v Concede quaesumus omnipotens Deus ut intercessio sanctae Dei genitricis Mariae sanctique Iohannis Baptistae 654, 7r, 13v Concede quaesumus omnipotens Deus ut quod beati Iohannis Baptistae sollemnia colimus 405, 218v Concede quaesumus omnipotens Deus ut sicut in nomine patris et filii divini generis intellegimus 353, 126r Confiteor tibi pater domine caeli et terrae siehe In illo tempore respondens Iesus dixit Confiteor tibi pater domine caeli et terrae quia abscondisti haec a sapientibus et prudentibus etc. (Mt 11,25). Confortatio fit multipliciter. Primo per restaurationem bonam sicut spiritus ex vino aromatico dicuntur confortari 748, 40v Consecrata est Seccoviensis ecclesia a venerabili in Christo patre et domino Hartmanno Brixinensi episcopo circa anno domini Mo Co XLo 293, 22r Constat in altari carnem de pane creari / illa caro Deus est qui dubitat reus est 167, 71vb Constitutio est ius relatum in scriptis redactum. Vel aliter est ius ubi non sequitur poena. Sed sanctio est ius ubi sequitur poena. Consultatio est in qua 155, 45va Consuetudo est regum terrae ut quando volunt aliqua seriosa bellica exercere 53, 249ra Consuetudo in mundo est quando aliquis usque in mortem in carcere cruciatur mittit ad amicos suos de quibus maxime confidit 78, 112ra Continuo oportet in medicina libros aphoristica brevitate collationes componere 453, 39r Contra aeris infectionem. Eius enim attractione membra principalia 748, 70v Contra debilitatem capitis naturalem 160, 1r Contra debilitatem oculorum equorum et macularum 160, 1r Contra inflationem colli equorum 160, 1r Contra scabiem equorum 160, 1r Contra vermes in intestivis equorum 160, 1r Contraria non fiunt in eodem. Cum ergo spiritus sanctus intrat in cor hominis tunc mala et peccata depellit 153, 137ra Conversus quidam in quodam coenobio cum monachis loquendo 308, 197rb Convertimini ad me … (Ioel 2,12). Unde Iohannes de Sancto Geminiano Videmus enim quod cum animal brutum sentit se vulneratum sollicite quaerit et procurat sanationis remedium 153, 60va Convertimini ad me et salvi eritis (Is 45,22). Quamvis divina miseratio multis variisque modis in scripturis sanctis omnis vocare dignata est 617, 63r Corda nostra quaesumus domine aeternae sapientiae splendor illustret 353, 19r 428 Corda nostra quaesumus – Cum turba plurima Corda nostra quaesumus domine passionum charismatum et gloriae rationalis 654, 11Ir Cordibus nostris quaesumus domine spiritum sanctum 353, 33r Corporis et sanguinis domini nostri Iesu Christi sacramentum sit 353, 56v Creaturarum consideratio pertinet ad theologos et philosophos 86, 1ra Credimus te Deum patrem a temetipso non ab alio nec factum nec creatum nec genitum te filium 353, 118v Cum aliquem pestis rapit eum taliter alterat et devorat ut velut incendium totum in brevi 748, 91r Cum aliquid petit suam confessionem audiri quem antea 532, 125v Cum appropinquasset Iesus Hierosolymis et venisset Bethfage ad montem Oliveti tunc misit duos discipulos etc. (Mt 21,1). Consuetudo est regum terrae ut quando volunt aliqua seriosa bellica exercere 53, 249ra – Hodie sancta mater ecclesia incipit celebrare adventum domini et hanc observantiam recepit a sanctis patribus 53, 1ra – Pro intellectu litterae notandum quod Iohannis 12 dicitur Quando Iudaei iam ex consilio Caiphae 53, 249va Cum audisset Iohannes in vinculis opera Christi mittens duos de discipulis suis ait illi etc. (Mt 11,2). Ex quo inter cetera multum provocat ad virtutes et virtuosorum hominum laudabilis conversatio 53, 25vb – Pro intellectu notandum quod secundum magistrum historiarum Iohannes arguebat Herodem propter Herodiadem quam a fratre suo receperat 53, 26ra Cum autem descendisset Iesus de monte secutae sunt turbae multae. Et ecce leprosus etc. (Mt 8,1). Christus dominus volens omnem dubitationem de sua divinitate a nobis excludere 53, 96va – Ecclesia instituit quod per totum annum cantantur et praedicantur Christi miracula 53, 110va – Quia postillatio verborum evangelii satis patet ex alio sermone facto de eodem. Ideo solummodo dicendum erit 53, 110vb – Sciendum quod dicit Haimo Mons in quo Deus sedet caelum intellegitur 153, 37va – Secundum Chrysostomum homilia 24 super Matthaeum Ne forte diceret populus apud se 53, 97ra Cum beatus Augustinus febribus laboraret et lecto decumberet 308, 14va Cum de febribus phlegmaticis iam tractandum est. Primo videndum est 123, 203r Cum de praesentis exilii [miseria] mortis transitus propter moriendi imperitiam 754, 18r Cum ergo sero factum esset die illa una sabbatorum etc. (Io 20,19). Hodie divina dies resurrectionis colitur in quo (!) perfecta requies designatur 53, 322rb Cum ex primo et principali nostrae regulae praecepto teneamur habere cor unum et animam unam 76, 17r Cum factus esset Iesus annorum duodecim ascendentibus illis in Ierusalem secundum consuetudinem etc. (Lc 2,42). Augustinus dicit in De consensu evangelistarum et magistri historiarum quod Herode mortuo 53, 76va – Christus dominus ne dum aetate virili sed etiam in pueritia sua se nobis ita exhibet ut ostendat quanta caritate nos diligat 53, 76ra – In praesenti enim evangelio instruit Maria parentes qualiter se debeant habere ad filios 153, 27va Cum ieiunasset quadraginta diebus … (Mt 4,2; vgl. Mc 1,13; vgl. Lc 4,1). Christus ieiunavit nobis in doctrinam et exemplum quia secundum Gregorium Omnis Christi actio nostra est instructio 153, 62va Cum ieiunatis nolite fieri sicut hypocritae tristes (Mt 6,16). Pro huius thematis introductione invenio tres scholas. Prima igitur schola fuit Moysii inesse figurali 597, 1ra Cum Martinus pergeret ad Candacensem dioecesim causa sedandae discordiae vidit mergos in flumine piscibus insidiantes 308, 28va Cum meae sit intentionis hoc in summario febrium 748, 2r Cum natus esset Iesus (Mt 2,1). Hodie triplex festivitas celebratur. Una est de adoratione magorum 617, 139r – Omnipotens Deus ex supereffluenti non ex operibus iustitiae quae fecimus 53, 66rb – Pro intellectu litterae notandum ut narrat Iosephus 13 libro Antiquitatum capitulo 14 regnum Iudaeorum fuit interceptum 481 annis et duos menses a captivitate 53, 71ra – Sicut Deus disposuit omnia sapienter sic fidei incrementum et suam humanitatem 53, 70va Cum non paucorum erga beneficia indulgentiarum mens errabunde vagetur 161, 1ra Cum quadam vice animum meum infelicem vanis ac transitoriis occupationibus assidue deditum ad meam recogitandam miseriam 532, 1r Cum sancto Lupo Senonensi archiepiscopo omnes qui de curia 308, 41rb Cum turba plurima conveniret et de civitatibus properarent ad Iesum dixit per similitudinem Exiit qui seminat seminare semen suum etc. (Lc 8,4). Beatus Bernardus in sermone De quadragesima dicit quod septem sunt impedimenta 53, 134ra – In praesenti evangelio dominus ostendit multorum auditorum Christi devotionem 53, 139ra – Praesens evangelium mea non indiget expositione quia summa veritas 53, 134va – Salvator noster Christus dominus saepius verbis suis nos admonet non solum mala fugere 53, 142rb – Secundum Bedam Satorem istum nullum melius quam Dei filium intellegere possumus 53, 142vb – Vera patientia quanta bona et quantas utilitates afferat homini possumus ex diversis scripturis sacris colligere 53, 138va Cum venerit filius – Deus pacis caritatisque 429 Cum venerit filius hominis in maiestate sua etc. (Mt 25,31). Pro huius thematis introductione invenio tres scholas scilicet Hebraicam, Graecam et Latinam. Prima schola est Hebraica in qua considero Salomonem facientem iustitiam 597, 8rb Cum venerit paraclitus quem ego mittam vobis a patre etc. (Io 15,26). Misericors pater qui est prope est omnibus invocantibus eum in veritate quia nos orphanos non relinquit 53, 344rb – Sicut in dominica proxima futura recolitur spiritus sancti missio 153, 129va Curatio. Aut enim intendimus sudorem provocare aut auferre 123, 11av Currite gentes undique et miramini erga vos caritatem Dei 125, 161ra; 165, 132v Da nobis domine continua omnium sanctorum tuorum veneratione laetari 405, 217v Da nobis quaesumus domine continua omnium sanctorum tuorum veneratione laetari 405, 218v Da nobis quaesumus domine vitiorum nostrorum flammas extinguere 405, 225v Da nobis quaesumus omnipotens Deus vitiorum nostrorum flammas extinguere 405, 219v Da quaesumus omnipotens et misericors Deus ut animae omnium fidelium defunctorum hic et ubique in Christo quiescentium 654, 41v Daemones enim frequenter plus solito ante et circa Christi adventum obsidebant homines 53, 239ra David dicit haec verba. Notandum est quod domus domini est quadruplex scilicet domus materialis 162, 2rb De cruce quando dominus ad cumulum portatur confestim ei genitrix portantes 353, 96v De constitutionibus d II et III per totum et d IV c 1 614, 270r De duabus ecclesiis noviter 125, 199va De dulcissimo nomine ipsius Mariae adhuc amplius videamus. Est notandum quod nomen huius dulcissimae virginis est a Deo revelatum 31, 58rb De duobus filiis summae deitatis (recte: veritatis) / Quos dedit ecclesiae pater pietatis / Quos eorum habeat plus auctoritatis 52, VDS De excelso misit ignem in ossibus meis et erudivit me R. (Lam 1,13). Quantum ad praesentem festivitatem possunt haec verba esse matris ecclesiae 617, 172r De nomine huius ineffabilis virginis adhuc est plus dicendum. Et est notandum quod haec virgo postquam Christum concepit reperitur in scriptura 31, 60va De parte autem animae. Iste est tertius liber partialis in quo dicitur de aliis animae potentiis 162, 71v De praesenti sollemnitate locuturus auribus vestrae caritatis dilectissimi amor venit ad memoriam 161, 6ra De prima parte evangelii dicendum est sed referendo ad nuptias corporales de quibus loquitur specialiter evangelium 53, 89vb De primo est sciendum duplicem esse ebrietatem vitandam … Secundo volens in religione proficere debet iuste cum fratribus vivere 617, 55r De quadam muliere dicebatur una vice casus supradictus fuisse 155, 7rb De tribus nuptiis legitur in evangelio 483, 67r Decrementa et incrementa paroxysmorum ad invicem aut sunt secundum quantitatem aut secundum tempora aut secundum qualitatem. Si secundum quantitatem aut longior 748, 1r Defende quaesumus domine Deus beata Maria semper virgine 405, 133v Descendit cum eis … (Lc 2,51). Augustinus Non solum Christi verba sunt nobis in documentum sed etiam Christi facta sunt nobis in exemplum 153, 30va Deserta cui inest miraculum 125, 199va Desidero domine mi certificari per te ex testimoniis legis et prophetarum et aliarum scripturarum 617, 117v Detur quid tunc si superest filius fratris ex uno parente coniuncti 155, 106v Deus a quo sancta desideria recta consilia et iusta sunt opera 405, 220r Deus amator omnium bonorum qui sanctissimum Augustinum confessorem 405, 218r Deus cui omne cor patet et omnis voluntas loquitur et quem nullum latet secretum 353, 35r; 405, 134v, 223v Deus cui omnia morientia vivunt et cui non pereunt moriendo corpora nostra sed mutantur in melius 654, 35v Deus cui proprium est misereri semper et parcere suscipe deprecationem 405, 207r Deus cuius misericordiae non est numerus suscipe propitius preces humilitatis nostrae 405, 129r; 14820, 86r Deus cuius unigenitus tuus pro nobis usque ad mortem oboediens 353, 70r Deus gratias tibi vera una trinitas: una et trina unitas te sanctam trinitatem 353, 118r Deus indulgentiarum domine da animae famuli tui (famulae tuae) cuius anniversarium depositionis diem et memoriam agimus refrigerii sedem quietis beatitudinem luminis claritatem 14820, 86r Deus indulgentiarum domine da fidelibus tuis quorum anniversarium depositionis diem et memoriam agimus 405, 129r Deus infinitae misericordiae veritatisque immensae propitiare iniquitatibus nostris et animarum nostrarum 654, 27r Deus largitor pacis et amator caritatis da famulis tuis 405, 133v Deus meus confundar et erubesco levare faciem meam 654, 17v Deus pacis caritatisque amator et custos da omnibus 405, 134r 430 Deus qui apostolo – Dicit Ecclesiasticus Deus qui apostolo Petro collatis clavibus 405, 218r Deus qui beatam Mariam Magdalenam dominicae resurrectionis gaudium sanctis apostolis innotescere voluisti 405, 218v Deus qui beatam virginem Margaretam ad caelos per martyrii palmam pervenire fecisti 405, 219r Deus qui beatissimam semper virginem Mariam in conceptu et partu virginitate servata triplici gaudio 353, 182v Deus qui beatum Nicolaum pontificem tuum innumeris decorasti miraculis 405, 219r Deus qui benedictae et gloriosae semper virginis genitricis filii tui Mariae corpus et animam sancti spiritus illustrationem perfecisti 353, 180v Deus qui caritatis dona per gratiam sancti spiritus cordibus fidelium infundis 405, 134v Deus qui corda fidelium sancti spiritus illustratione docuisti 353, 38r; 405, 219r, 224r Deus qui crucem sanctam ascendisti et mundi tenebras illuminasti 405, 216v Deus qui de beatae Mariae virginis utero verbum tuum angelo annuntiante carnem suscipere voluisti 405, 211r Deus qui dedisti legem Moysi in summitate montis Sinai et in eodem loco per sanctos angelos 405, 219r Deus qui diem discernis a nocte 405, 129v Deus qui diligentibus te misericordiam tuam semper impendis et a servientibus tibi in nulla 405, 134r Deus qui ecclesiam tuam pretiosi corporis et sanguinis domini nostri Iesu Christi mysterio 353, 63v Deus qui fidelium tuorum deprecationem semper exaudis praesta mihi indigno peccatori 353, 12v Deus qui miro ordine angelorum ministeria hominumque dispensas concede propitius 405, 217r Deus qui nobis sub sacramento mirabili passionis tuae memoriam reliquisti tribue 353, 60v; 405, 219v Deus qui nos annua beati Floriani martyris tui sociorumque eius gloriosa passionis sollemnitate laetificas 14820, 86r Deus qui nos beatae Mariae semper virginis et beatorum spirituum patriarcharum 654, 7Iv Deus qui nos concedis omnium sanctorum memoriam agere 353, 149r Deus qui nos concedis omnium sanctorum tuorum merita venerari 654, 7v Deus qui nos hodie beatae virginis tuae annua sollemnitate laetificas 154, 4r Deus qui nos miseros peccatores perire non sinis praesta quaesumus ut angelum mihi deputatum 353, 9r Deus qui nos patrem et matrem honorare praecepisti miserere quaeso clementer animabus patris mei et matris meae omniumque parentum 654, 87r Deus qui novo stellae signo novum filium regibus ostendisti praesta quaesumus ut intercedente benedicta et gloriosa semper virgine 353, 176r Deus qui omnipotentiam tuam parcendo maxime et miserando manifestas multiplica super nos misericordiam tuam 353, 45v Deus qui per coaeternam sapientiam hominem cum non esset condidisti 353, 18r, 20r, 23v Deus qui populo tuo beatum Augustinum aeternae salutis ministrum constituisti 405, 222r Deus qui post baptismi sacramentum secundam abolitionem peccatorum eleemosynis 405, 134r Deus qui salutis aeternae beatae Mariae virginitate fecunda humano generi praemia praestitisti 353, 146r; 405, 211v Deus qui summa providentia humano generi supernorum spirituum praesidia subministras 353, 5v Deus qui tribus pueris mitigasti flammas ignium concede propitius 154, 8r; 252, 24r; 405, 225r Deus qui unicum filium tuum ut servum redimeres sponte in mortem crucis tradidisti 654, 45v Deus qui unigeniti filii tui domini nostri Iesu Christi pretioso sanguine humanum genus redimere dignatus es 654, 50v Deus qui unigeniti filii tui pretioso sanguine vivificae crucis vexillum sanctificare voluisti 405, 221r Deus qui virginalem aulam beatae Mariae virginis in qua habitares eligere dignatus es 353, 30v Deus sub cuius oculis omne cor trepidat omnesque conscientiae contremiscunt 654, 30v Deus veniae largitor et humanae salutis amator quaesumus clementiam tuam 405, 128v, 129r; 654, 87r Deus vitae dator humanorum corporum reparator qui a peccatoribus exorari voluisti exaudi quaesumus preces nostras 654, 36r Devotus sit in choro discretus in capitulo sobrius in refectorio disciplinatus in omni loco 145, 314va Dialectica est ars artium scientia scientiarum ad omnia methodorum principia habens viam. Sola enim dialectica probabiliter disputat de principiis omnium artium 605, 1ra Dic ut sedeant hi duo filii mei etc. (Mt 20,21). Filius Dei volens loqui Iudaeis dixit haec verba quae etiam ego possum dicere. Mea doctrina non est mea sed eius 617, 178v Dicit beatus Augustinus quod duplex est civitas 162, 8vb Dicit beatus Iohannes in suo evangelio quod dominus interrogavit beatum Petrum de sua dilectione dicens 617, 175v Dicit Ecclesiasticus (39,3) occulta Proverbiorum exquirit sapiens et in absconditis parabolarum 156, 1r Dicite filiae Sion – Domine Iesu Christe 431 Dicite filiae Sion Ecce rex tuus venit tibi mansuetus etc. ubi supra (Mt 21,5) et originaliter Zachariae 9 capitulo (9). Secundum doctores adventus domini agitur per quattuor hebdomadas ad denotandum quod est quadruplex adventus filii Dei 53, 1va Dicitur vulgariter Bonum ferrum trahit homini argentum de bursa 617, 188v Dicto de restitutione fienda damnificatis in rebus exterioribus nunc restat dicere de restitutione fienda in bonis animae et corporis 53, 227ra Dictum est prius quomodo dulcissima virgo Maria est gloriose salutata in hoc verbo Ave. Nunc dicendum est quomodo gratiose est adornata 31, 69ra Digitos in aures surde (!) et tandem rogaverunt tam ut imponeret sibi manum 177, 134rb Dilecto sibi in Christo socio et amico N. clerico de tali loco verae sapientiae et augmentum continuum vitae praesentis. Iste liber cuius subiectum est ens mobile contractum ad naturam secretorum mulierum 155, 123r Diligite inimicos vestros (Mt 5,44). Pro huius thematis introductione tres scholas reperio scilicet naturae, scripturae, gratiae. Primo dico quod invenio scholam naturae 597, 4ra Diligite iustitiam qui iudicatis terram. Circa istum librum qui liber Sapientiae nuncupatur sunt in principio tria notanda 200, 2va Dirigere sanctificare et regere digneris domine sancte pater omnipotens aeterne Deus 405, 128r Discite a me quia mitis sum … (Mt 11,29). Aristoteles In sempiternis nihil violentum. Qui ergo alte in caelo vult 153, 46vb Dissipavit bona sua vivendo luxuriose (Lc 15,13). Pro huius thematis introductione tres invenio scholas scilicet Medeae, Dyodonis, Ysiphilae. Prima ergo schola fuit Medeae nam Medea fuit filia Ochis (!) regis qui regnavit in insula Colchide 597, 25va Distinctio prima Christus est nobis prope quattuor modis 174, 4v Distribuit discumbentibus (Io 6,11). Pro huius thematis introductione reperio tres scholas mirificam, munificam, magnificam. Prima schola est mirificum (!) et haec schola est Dei. Prope enim Deus qui fecit mirabilia 597, 32vb Dividitur autem praesens opusculum in tres partes in quarum prima agitur de sacramentis 550, 2ra Divina dispositione et providentia ab aeterno Deus piissimus disposuit postquam Iudaicus populus 53, 333rb Divina scriptura istam habet proprietatem quod in una sui parte fideles admonet lacrimari 53, 304vb Dixit angelus ad virginem de caelo missus. Et benedictus fructus ventris tui. Adicit Elisabeth spiritu prophetico inspirata 31, 49ra Dixit Iesus turbis Iudaeorum et principibus sacerdotum Quis ex vobis arguet me de peccato si veritatem dico quare non creditis mihi etc. (Io 8,46). Quod quilibet homo et maxime Christianus debeat fugere lites et contentiones 53, 246ra Doctrinarum autem animae. Capitulum tertium in quo determinatur de potentiis animae 163, 50v Domina mea sancta Maria perpetua virgo virginum mater summae benignitatis et misericordiae deprecor te 654, 110r Domine Deus pater omnipotens qui nos ad principium huius diei pervenire fecisti 405, 127v Domine Deus pater omnipotens qui consubstantialem et coaeternum tibi ante omnia saecula ineffabiliter 405, 207v Domine Deus virtutum qui collapsa restauras et restaurata conservas 353, 193v Domine Iesu Christe absolve me per sanctam captivitatem tuam ob omnibus vitiis 405, 211r Domine Iesu Christe clementissime qui pro nobis peccatoribus mortem subiens temporalem nec materno dolori 353, 101v Domine Iesu Christe fili Dei vivi mundi creator et redemptor qui dixisti 405, 220v Domine Iesu Christe fili Dei vivi pone amaram mortem et passionem tuam inter animas nostras et misericordiam et districtum iudicium 14820, 86r Domine Iesu Christe fili Dei vivi qui dixisti nolo mortem peccatoris sed ut magis convertatur et vivat quaeso pone amaram mortem 353, 143v, 146v, 149v, 152r, 153Iv, 158r Domine Iesu Christe fili Dei vivi qui es verus et omnipotens Deus splendor et imago patris vita aeterna 405, 207v Domine Iesu Christe fili Dei vivi qui hora completorii sepultus es et a mulieribus planctus es et lamentatus 353, 92r Domine Iesu Christe fili Dei vivi qui hora diei nona pendens in cruce et clamans voce magna te derelictum dixisti 353, 85v Domine Iesu Christe fili Dei vivi qui hora diei prima permisisti te duci in consilio Iudaeorum 353, 79r Domine Iesu Christe fili Dei vivi qui hora diei tertia fuisti ab Herode illusus 353, 81v Domine Iesu Christe fili Dei vivi qui hora matutina stans ligatus 353, 77r Domine Iesu Christe fili Dei vivi qui hora vesperarum cenasti cum discipulis tuis turbatus spiritu unum ex eis te traditurum 353, 89r Domine Iesu Christe fili Dei vivi qui pretioso sanguine tuo mundum redemisti 654, 47r Domine Iesu Christe fili Dei vivi qui pro nobis in cruce moriens dulcissimi lateris 353, 66r Domine Iesu Christe fili virginis et matris Mariae qui hora matutinali pro salute humani generis ex Maria 353, 101v 432 Domine Iesu Christe – Dum temptationes diaboli Domine Iesu Christe fili virginis Mariae salus mundi qui hora diei nona crucifixus manibus et pedibus verberibus et flagellis 353, 108v Domine Iesu Christe fili virginis Mariae salus mundi qui hora diei prima Pilato praesidi fuisti praesentatus 353, 104r Domine Iesu Christe fili virginis Mariae salus mundi qui hora diei sexta in cruce alta clavis durissimis affixus fuisti 353, 107r Domine Iesu Christe fili virginis Mariae salus mundi qui hora diei tertia coram Pilato praeside purpura indutus fuisti 353, 105v Domine Iesu Christe fili virginis Mariae salus mundi qui hora diei vespertina pressuris innumeris nimis fatigatus 353, 111v Domine Iesu Christe iudex vivorum et mortuorum qui ob hoc irasceris ut subvenias ad hoc minaris 654, 49r Domine Iesu Christe qui ante iudices legatis manibus stare voluisti 405, 212r Domine Iesu Christe qui dum hora sexta pro redemptione mundi crucis ascendisses lignum 353, 190v Domine Iesu Christe qui hora diei sexta ante crucem fuisti denudatus 353, 83v Domine Iesu Christe qui in cruce suspensus es pro nostra redemptione tuum dulcissimum spiritum in manus Dei patris 405, 224v Domine Iesu Christe qui in ultima cena tradidisti discipulis tuis 353, 68r Domine Iesu Christe qui pro nobis peccatoribus in cruce agonizans 353, 103v, 105v Domine Iesu Christe qui proprio sanguine tuo mundum redemisti redime me ab omnibus peccatis et neglegentiis 405, 215r Domine Iesu Christe qui propter me mortem pati voluisti 405, 214r Domine Iesu Christe rogo te per amaram extensionem omnium membrorum et nervorum tuorum 405, 213v Domine Iesu Christe sanctifica me per spineam coronam et per sacra vulnera 405, 213r Domine Iesu Christe virginis fili dulcissime qui cum proditore cenasti 353, 111r Domine nonne bonum semen seminasti etc. (Mt 13,27). Plures sunt agri in quibus seminat 177, 44vb Domine sancte pater omnipotens sempiterne Deus famulos tuos maiestati tuae subiectos per unicum filium tuum 353, 125r Domine sancte spiritus qui coaeternus coaequalis et consubstantialis patri filioque exsistens ab eis inenarrabiliter 405, 208r Domine si fuisses hic frater meus non fuisset mortuus (Io 11,21). Pro huius thematis introductione reperio tres scholas scilicet elementalem, spiritualem, iudicialem. In his tribus scholis legitur mors. Prima ergo schola est elementalis quia composita originaliter ab his cum omne compositum resolvi 597, 34vb Domini Iesu felix anima divinitati personaliter unita humanae infirmitatis defectus habuit mihi tristis fuit 353, 185r Dominus Deus creavit hominem immo et totum mundum propter suam bonitatem 53, 124ra Dominus Deus non invitat solum cottidie per suos nuntios praedicatores sed etiam per se ipsum 53, 119va Dominus dicit in evangelio Paenitentiam agite appropinquabit enim regnum caelorum (Mt 3,2). Petrus apostolus dicit Paenitentiam agite et baptizetur 181, 41rb Dominus Iesus Christus et magister noster pretiosum nostrae redemptionis pretium 53, 272rb Dominus tecum (Lc. 1,28). O Maria dominus tecum te associans tecum te custodiens tecum te glorificans 31, 82ra – Secundum ordinem naturalem in mundo corpora magis conformia in natura sint magis propinqua in situ 31, 79vb Domus mea domus orationis vocabitur (Mt 21,13). Pro huius thematis introductione invenio tres scholas scilicet Belzebub in Babylonia, Iani in Roma, Pharisaeorum in Ierusalem. Prima schola fuit in magna civitate Babyloniae in qua tractabatur 597, 9va Dona ferunt proceres colitur quibus inclitus heres 308, 228va Dubitatur numquid aliquid peccatum sit tam grande 165, 114r Dubitatur utrum scientia naturalis est scientia de omnibus rebus 168, 1r Ductus est Iesus in desertum a spiritu ut temptaretur a diabolo etc. (Mt 4,1). Ex quo omnia tempora tempus suum habent ut dicit sapiens Ecclesiastes 3 Est tempus flendi 53, 159ra – Ex quo sequitur assumpsit eum diabolus etc. Dubitatur a quo spiritu ductus sit in desertum 53, 159vb – Glossa dicit Christus non permisit se temptari nisi post baptismum 53, 164va – Pro intellectu dubitatur quale fuit illud desertum ad litteram 53, 174va – Sicut dicit Gregorius homilia 2 super evangelia Miracula domini et salvatoris nostri sic accipienda sunt 53, 174ra – Sicut in bello corporali securius pugnatur et citius obtinetur triumphus contra hostes 53, 169rb – Tota summa huius evangelii est quod Christus voluntarie ivit ad desertum locum 53, 170ra – Totam vitam nostram plenam fecit Deus temptationibus et etiam rationabiliter Deus permittit electos suos temptari 53, 164ra Dum mors grassatur puero fuga tuta paratur 308, 228va Dum temptationes diaboli ipsum nimio molestarent 308, 47va Duo enim manifestantur – Equus calidae naturae 433 Duo enim manifestantur in evangelio praesenti scilicet passio Christi et illuminatio cuiusdam caeci 153, 55rb Duodecim sunt signa scilicet aries taurus … Notandum quod in his duodecim signis sol perficit cursum suum 162, 4va Ea facta quae dubia sunt ut quo animo in meliorem partem interpretemur 614, 282r Ecce ascendimus Hierosolymam … (Lc 18,31; Mt 20,18; Mc 10,33). Duo enim manifestantur in evangelio praesenti scilicet passio Christi et illuminatio cuiusdam caeci 153, 55rb Ecce ego mitto angelum meum qui praeparabit viam ante faciem tuam (Mal 3,1). In verbis istis quattuor notantur. Primum est quod dominus dicit Ecce ego mitto angelum meum etc. Secundum est quod dicit 78, 29rb Ecce evangelizo vobis … (Lc 2,10). Nota quod inter omnia festa laetabunda totius anni hoc festum nativitatis est tanto laetius 153, 1va Ecce filiae Ierusalem quas delicias vobis praeparavi 483, 76v Ecce magi veniunt … (Mt 2,1). Hodiernae diei festivitas magna est. Unde merito dicitur apparitio 153, 23ra Ecce nunc tempus acceptabile ecce nunc dies salutis (2 Cor 6,2). Pro huius thematis introductione tres scholas reperio et fortunae, naturae, rationalis creaturae. Prima schola scilicet naturae dicit quod omnia naturalia 597, 6rb Ecce video caelos apertos (Act 7,56). Prosequendo sermonem iam inceptum tunc sciendum quod quanto caelum aperitur per intensam contritionem 153, 9rb Ecclesia instituit quod per totum annum cantantur et praedicantur Christi miracula 53, 110va Ecclesiae tuae quesumus domine preces placatus admitte 654, 65r Ecclesiam tuam quaesumus domine benignus illustra ut beati Iohannis apostoli tui et evangelistae illuminata doctrinis 405, 217v, 218v Ego dixi miserere mei domine. Sana animam meam quia peccavi tibi. Convertere domine 353, 53v; 405, 223v Ego dominus exaltavi lignum humile et humiliavi lignum sublime (Ez 17,24). In his verbis agitur de duobus lignis. Primo de ligno salvationis in quo homo fuit vivificatus 617, 187Ir Ego sum nolite timere (Mc 6,50). Pro huius thematis introductione tres scholas reperio scilicet Epicureorum, Platonicorum, Hypotheticorum. Sed in omnibus istis scholis doctrina legitur quae secundum aliquam partem merito nobis timorem 597, 5rb Ego sum pastor bonus. Bonus pastor animam suam dat pro ovibus suis (Io 10,11). Licet multae sunt causae in particulari quare lites et turbationes surgunt inter homines in mundo 53, 325ra – Hic Christus seipsum commendat. Unde hic est dicendum quando quis potest se licite commendare et quando est veniale et quando est mortale 153, 109vb – Sciendum quod per pastorem in praesenti evangelio intelleguntur praelati et per oves intelleguntur subditi 153, 107vb Ego veniam et curabo eum (Mt 8,7). In his verbis notatur magna humilitas Christi qui servum centurionis dignatus est visitare 153, 39rb Egredietur virga de radice Yesse (Is 11,1). Legitur de quinque virgis laudabilibus per quas beata virgo Maria merito figuratur 617, 146v Egregius praedicator et apostolus invitans nos ad opera virtutum. In istis verbis tria facit. Primo allegat temporis opportunitatem 174, 168ra Egressus Iesus cum discipulis suis trans torrentem Cedron ubi erat ortus etc. (Io 18,1). Quando enim occurrat festum alicuius sancti tunc consuevimus eius legendam et passionem ut Laurentii Petri etc. Sic enim hodie est flendum 174, 156ra – Sapiens Ecclesiastes 3 (1 und 4) dicit quod omnia tempora tempus habent. Est tempus flendi et tempus ridendi 53, 280rb – Sicut enim tunc vos scitis, quod mos praedicatorum est in principio sermonum praedicatorum ad beatam virginem recurrere 174, 156Ira Egressus Iesus perambulabat Iericho (Lc 19,1). Zacchaeus interpretatur iustus sed quia ille iustus non erat ideo dominum videre non poterat. Sunt autem tres causae quare dominum videre non poterat. Primo propter divitias 162, 4ra Egressus Iesus secessit in partes Tyri et Sidonis etc. (Mt 15,21). Inter cetera quae reconciliant peccatorem Deo postquam in peccatum mortale corruit est recognitio 53, 236vb – Pro intellectu notandum sicut habetur Mt 14 Postquam Christus transfretavit per mare Tiberiadis venit Capharnaum 53, 237rb Emitte quaesumus domine lucem tuam in corda nostra ut mandatorem 405, 41r Eo tempore in Lamon sanctissimus monachus 308, 42ra Eorum quae. Iste est liber secundus Physicorum in quo Aristoteles determinat de principiis doctrinae 168, 9r Equus calidae naturae iudicatur sed temperatae calor ostenditur per levitatem 160, 4rb 434 Erant duo socii – Ex quo in praesenti evangelio Erant duo socii simul coniuncti unus quaesivit alterum quis eorum remotius ambulasset. Dixit primus Fui in provincia 125, 311v Erat Iesus eiciens daemonium et illud erat mutum etc. (Lc 11,14). Chrysostomus et Theophilus dicunt et habetur Matthaei 12 Ille daemoniacus erat mutus 53, 240va – Daemones enim frequenter plus solito ante et circa Christi adventum obsidebant homines 53, 239ra – Iohannes de Sancto Geminiano Videmus naturaliter quod lux habet expellere tenebras. Christus autem est lux 153, 69ra – Quam detestabile sit vitium invidiae et fugiendum patet propter multa mala 53, 240ra – Secundum quod dicit Beda in homilia de Christi transfiguratione ut patet Matthaei 17 Salvator humani generis Christus dominus 53, 238va Erat Ioseph et Maria mater Iesu mirantes super his quae dicebantur de illo etc. (Lc 2,33). Augustinus in quodam sermone de Christi nativitate qui incipit Adiuvet nos humillimos 53, 57va – Evangelium praesens tangit primo de parentibus salvatoris id est patre et matre 53, 58rb Erudi quaesumus domine plebem tuam 194, 1r; 726, 1r Erunt signa in sole luna et stellis etc. (Lc 21,25). Quod multum terribile sit extremum iudicium videtur posse trahi ex his quae in proximo sermone dixi 53, 11rb – Christus dominus per scripturas nobis valde incutit timorem de extremo iudicio 53, 6va – In praesenti evangelio monet nos dominus quatenus praeparemus nos ad istam ultimam rationem quam oportet nos facere in extremo iudicio 53, 7ra Est amor ordinatus videlicet ut amet 165, 42ra Est discretio testantisque suprema voluntas actio 155, 122ra Est dubitatio de monialibus occulte corruptis quia in quibusdam monasteriis consuetudo est quod solae virgines 155, 10ra Est enim semen verbum Dei quod Christus de caelo attulit in terra seminavit 153, 54ra Est puer unus hic qui habet quinque panes … (Io 6,9; vgl. Mt 14,17). Iohannes de Sancto Geminiano dicit quod totum officium praesentis diei dominicae continet quasi quaedam nova et laeta 153, 71vb Est triplex typus rigor oripilatio frigus 748, 176ra Estote imitatores Dei sicut filii carissimi (Eph 5,1). Pro huius thematis introductione invenio tres scholas scilicet dominorum, generatorum, magistrorum. Prima schola fuit dominorum et in ista et in aliis legitur quod omnes intendunt sibi assimilare 597, 27ra Estote misericordes sicut et pater vester misericors est (Lc 6,36). Nota in praecedenti evangelio dominicali dicitur quod publicani et peccatores 174, 205va Et accedens temptator dixit ei Si filius Dei es … (Mt 4,3). Sciendum quod Christus voluit temptari ut ostenderetur quod nos spiritualiter pugnare oporteret 153, 65rb Et cum ieiunasset 40 diebus et 40 noctibus postea esuriit (Mt 4,2). Sancti patres primitivi Christianae ecclesiae instituerunt ieiunium fidelibus observandum 53, 176vb Et dedimur a Musis quae per Bocacium XI libro De genealogia Deorum gentilium 776, 23r Et in evangelio hodierno sicut scriptura sacra Veteris et Novi Testamenti persuadet fideles ut diligant et amplexentur humilitatem 53, 30ra Et quaerebat videre Iesum quis esset (Lc 19,3). Haec verba scribit Lucas de quodam Iudaeo qui vocabatur Zachaeus qui desiderabat videre filium Dei 162, 7va Et si venenum multipliciter accipi possit hic per venenum id solum intellego quod qualitercumque humano corpori 748, 122r Et tunc videbunt filium hominis venientem in nube cum potestate magna et maiestate (Lc 21,27). Sicut valde consultum est servo procuratori alicuius potentis domini qui numquam rationem fecit 53, 19rb – In istis verbis describit salvator adventum suum ad iudicium extremum 53, 20ra Et ut tanti beneficii iugis in nobis maneret memoria corpus suum in cibum 353, 57v Et veniat super nos misericordia tua domine 654, 64r Evangelica clamat historia. Sine intermissione orate hoc est semper quidem orat qui semper bene agit 617, 1r Evangelium praesens tangit primo de parentibus salvatoris id est patre et matre 53, 58rb Ex praemissis patet quod quam cito homo moritur qui adhuc stat in innocentia baptismali 153, 152va Ex quo dominus noster Iesus Christus in verbis praemissis invitat nos ad orandum tunc hic quaeritur 153, 123rb Ex quo evangelium hodiernum mentionem facit de matrimonio et de nuptiis in quibus Christus Deus et homo cum benedicta matre 153, 35va Ex quo hodie est ultima dies celebris praesentis festivitatis paschae et modo homines redire debent ad pristinos labores suos 153, 101ra Ex quo hodie peragitur festum illorum puerorum innocentium qui ab iniquo Herode interfecti sunt 153, 14ra Ex quo iam illis sacris diebus peragitur missio spiritus sancti tunc aliquid de donis spiritus sancti pro praesenti sermone dicendum est 153, 141rb Ex quo in praesenti evangelio fit mentio de patrefamilias qui conduxit operarios 153, 50ra Ex quo in praesenti evangelio – Filius Dei volens loqui 435 Ex quo in praesenti evangelio fit mentio de puero nobis recenter nato. Tunc pro praesenti sermone tria sunt dicenda. Primo quare Christus parvulus factus est 153, 15vb Ex quo in verbis praemissis fit mentio de partu mulierum. Tunc in praesenti sermone tria sunt dicenda 153, 112vb Ex quo inter cetera multum provocat ad virtutes et virtuosorum hominum laudabilis conversatio 53, 25vb Ex quo omnia tempora tempus suum habent ut dicit sapiens Ecclesiastes 3 Est tempus flendi 53, 159ra Ex quo sequitur assumpsit eum diabolus etc. Dubitatur a quo spiritu ductus sit in desertum 53, 159vb Ex quo spiritus sanctus est fons totius dulcedinis tunc quaeritur quare Deus quandoque dilectis suis subtrahit in praesenti consuetam gratiam 153, 139ra Ex quo tota Christi vita et conversatio nostra est instructio 53, 63ra Ex testimoniis quattuor evangeliorum et exemplo Hieronymi contra Helvidium Sancta Maria mater domini et Maria mater Iacobi Alphaei et Ioseph et mater filiorum Zebedaei scilicet maioris Iacobi et Iohannis evangelistae tres sorores fuerunt 293, 23r Exaudi domine Iesu Christe preces servorum tuorum qui cum Deo patre et spiritu sancto 405, 7v Exaudi domine preces nostras quas in sancti Wolfgangi confessoris tui atque pontificis sollemnitate deferimus 14820, 85v Exaudi domine preces servi tui ad te clamantis qui in trinitate perfecta vivis et regnas Deus 353, 185r Exaudi domine preces servorum tuorum ut animae fidelium defunctorum a poenis liberentur infernorum 654, 35r Exaudi nos Deus salutaris noster 405, 167r Exaudi nos domine sancte pater omnipotens aeterne Deus invocationem omnipotentis filii tui 353, 130r Exaudi nos misericors Deus et mentibus nostris sapientiae tuae lumen ostende 353, 21v Exaudi quaesumus domine preces supplicum et confitentium tibi 654, 25r Excita quaesumus domine in ecclesia tua spiritum cui beatus Augustinus confessor tuus et pontifex servivit 405, 221v Exemplum de quodam rege qui habuit duos camerarios valde invidos 308, 219ra Exemplum legitur quod quidam pauper manibus laborabat 308, 201vb Exiit edictum a Caesare Augusto (Lc 2,1). Secundum historiam post mortem Iulii Caesaris qui a poetis commendatur 53, 47vb – Nota Octavianus imperator dictus est Augustus primus quia regnum Romanorum notabiliter auxit 53, 43va – Sicut dicit Bernardus in sermone de epiphania Priusquam appareret Dei humanitas latebat 53, 47va – Sicut omnipotens Deus verax est in praemissis Ita et misericors 53, 43rb Exiit qui seminat … (Lc 8,5). Ubi sciendum quod seminator est Deus omnipotens qui exiit seminare semen suum id est genus humanum creando 153, 51vb Exorcizo te creatura salis in nomine patris omnipotentis et in caritate domini nostri Iesu Christi et in virtute spiritus sancti 726, 71r Expurgate vetus fermentum etc. (1 Cor 5,7). Hodiernam paschalem videlicet festivitatem inter ceteras festivitates Christi praecipuam 53, 313vb Extendit Noe manum et comprehensam columbam in arcam intulit (Gn 8,9). Hic tria possunt notari scilicet Mariae assumptio cum dicitur Extendit Noe manum secundo festivitas venerabilis Mariae cum dicitur Apprehensam columbam tertio assumptionis securitas cum dicitur In arcam 617, 182v Fac nos quaesumus domine sanctae Mariae semper virginis subsidiis attolli 654, 64v Facite dignos fructus paenitentiae (Mt 3,8; Lc 3,8). Ita hortatur nos praedicator paenitentiae beatus Iohannes Baptista in deserto praedicans baptismum paenitentiae Matthiae 3 et Lucae 3 quod specialiter nobis dicitur hoc sacratissimo ieiuniorum tempore legimus 53, 227vb Famulorum tuorum quaesumus domine delictis ignosce ut qui tibi placere 353, 151v; 405, 214v, 221v Febris secundum Avicennam quarto canonis in principio est calor extraneus accensus in corde et ab eo procedens mediantibus spiritu et sanguine … Et talis calor extraneus quandoque est in spiritibus, quandoque in umoribus 123, 195r Fecit Moyses labrum aeneum de speculis mulierum quem plenum aqua posuit 293, 1r Felices sancti Dei omnes qui pertransistis pelagus mortalitatis et pervenire 654, 2v Fert paranymphus ave quo mundus solvitur ave 308, 228rb Fertur quoque quod dum vir quidam sancto Augustino valde devotus in spiritu raptus 308, 16va Festina quaesumus domine Iesu festina ne tardaveris et auxilium nobis supernae virtutis impende 353, 178v Fideles tuos quaesumus domine ab ira tua confugientes ad te paterna recipe pietate 654, 32v Fidelium Deus omnium conditor et redemptor animabus famulorum famularumque tuarum omnium fidelium 405, 129r; 654, 87v, 92v Filium tuum omnipotens pater ante saecula gemitum invocantes exoramus 353, 128r Filius carnalis dicit confessori suo se duxisse in uxorem illam quam pater suus baptizaverit 155, 9vb Filius Dei volens loqui Iudaeis dixit haec verba quae etiam ego possum dicere. Mea doctrina non est mea sed eius 617, 178v 436 Fons crevit – Has horas canonicas Fons crevit in flumen magnum (Est 10,6). Ibi dic historiam qualiter Mardochaeus somniavit duos dracones 617, 147r Fons vitae lassus solus sitit essibus assus 308, 228vb Formavit Deus hominem ad imaginem et similitudinem suam 7, 66ra Fragilitatem nostram quaesumus domine benignus respice et saporem nobis aeternae sapientiae 353, 25v Frater Ruodbertus eius monachus et secundum carnem 308, 62ra Frequenter fit aurum potabile sic et confert pestilentiatis. Recipe auri optimi 748, 64v Fuit quidam rusticus qui cum iret 308, 200vb Fuit quidam vir lascivus valde contigit eum 308, 198rb Galienus secundo de crisi dicit quod si videris urinam aegri tendentem 748, Iv Gaude felix mater cum tibi per angelum Dei filius nuntiatur cum pudicus alvus tuus spiritu sancto fecundatur 353, 166v Gaude virgo quae de caelis iuxta vocem Gabrielis concepisti filium. Gaude mater Iesu Christi quia virgo peperisti creatorem omnium 353, 182v Gaudete et exsultate omnes sancti quoniam merces vestra multa est in caelo (Lc 6,23). Verba proposita leguntur in evangelio hodierno et per ea insinuatur nobis gloria praemium omnium sanctorum quorum hodie festivitas celebratur 617, 189r Gloriosa passio Christi perducat nos ad gaudia paradisi 353, 71r Glossa dicit Christus non permisit se temptari nisi post baptismum 53, 164va Glossa non fuit satis evangelistae dicere gavisi sunt sed addit valde magno gaudio 153, 25vb Glossa semper quam non habet. Unde hic est dicendum quomodo homo debet iustus inveniri si vult salvari 153, 119vb Glossa semper quod habet. Unde hic est dicendum quomodo quis committit septem peccata mortalia 153, 119vb Glossarum diversitas intelligentiam textus nec numquam obtenebrat 253, 1ra Graecorum studia nimiumque diuque secutus 327, 20ra Grana quater quinque scrupuli pro pondere summae 748, 212vb Grata tibi sit domine haec oblatio plebis tuae quam tibi offerimus in honore sanctae N virginis tuae ut cunctis proficiat ad salutem 154, 4r Gratia plena (Lc 1,28). Dictum est prius quomodo dulcissima virgo Maria est gloriose salutata in hoc verbo Ave. Nunc dicendum est quomodo gratiose est adornata 31, 69ra – Notandum quod sicut sunt septem utilitates sive efficaciae quas gratia confert animae de quibus dictum est in sermone praecedenti dicendum est 31, 71va – Prius dictum est quod gratia habet septem efficacias quas in anima facit et quod sunt septem praeparationes quibus homo ad gratiam se disponit 31, 73rb – Prius dictum est quomodo Maria fuit gratiose salutata quod notabatur in hoc verbo Ave 31, 75vb Gratias tibi agimus domine sancte pater omnipotens aeterne Deus qui nos de transacto noctis spatio ad matutinas horas perducere dignatus es 405, 51r, 65v Gratias tibi ago omnipotens Deus qui me dignatus es custodire in hac nocte 405, 208r Grex cum pastore pacis laetatur amore 308, 228va Hae duae fuerunt ultimae sorores ex parte matris cum virgine gloriosa 53, 299vb Haec circa officium curatorum breviter a me scripta sunt 550, 88ra Haec est dies quam fecit dominus … (Ps 117,24). Sciendum quod praesens festivitas praecellit alias festivitates in sollemnitate et iucunditate 153, 89va Haec est vita virginis Mariae gloriosae / Summi Dei filii matris generosae … Maria sibi regulam statuit vivendi / et sub quodam ordine Deo serviendi 166, 51va Haec ita cogitans menti occurrit illa vocatio trina apostolorum qui primo ad notitiam 532, 1r Haec sunt virtutes aquae ardentis vel vitae. Est enim clarior quam aqua rosacea et quia plus est secure 748, 53ra Haec verba canit ecclesia per illud sacrum tempus paschale usque ad ascensionem 153, 91vb Haec verba licet ad litteram sint dicta de agno paschali cuius immolatione 53, 251va Haec verba locutus est dominus Deus et ante passionem suam post cenam 153, 109vb Haec verba possunt exponi de beata Maria Magdalena quae intellegitur per arcum caelestem propter tria. Primo ratione materiae 78, 69vb Haec verba scribit Lucas de quodam Iudaeo qui vocabatur Zachaeus qui desiderabat videre filium Dei 162, 7va Haec verba sunt beati Petri ad commendationem et dignitatem sacerdotalem. In verbis praemissis possumus notare triplicem gradum sine excellentiam cuiuslibet sacerdotis. Primo eius dignitatem 597, 58ra Haereticus quidam illud gloriosum corpus dicens publice 125, 200ra Has horas canonicas Maria mater pia tuae laudi refero cordis symphonia 353, 161v Helleborus niger decoctus – Illud est prius 437 Helleborus niger decoctus in pomo sub prunis datus in scrupulis II optime purgat tempore pestilentiae 748, 69v Heres est. Venite occidamus eum (Mt 21,38). Pro huius thematis introductione tres scholas invenio scilicet Polestinoris, Mesabellis, Pignaleonis. Prima schola fuit Polestinoris qui fuit rex cuiusdam insulae 597, 24rb Hic dicendum est de caritate Dei quam habet ad nos et etiam de caritate quam nos ad ipsum habere debemus 153, 109vb Hic est dicendum quare Deus permittit aliquos bonos homines ita acerbissime tribulari in praesenti 153, 115rb Hic est dicendum quare gaudium mundi est fugiendum et quando est mortale 153, 115va Hic est vere salvator mundi (Io 4,42). Pro huius thematis introductione invenio tres scholas scilicet lapidum, herbarum, verborum. Prima ergo schola est lapidum et in illa legitur magna virtus. Nam videmus quod carbunculus 597, 30va Hic Christus seipsum commendat. Unde hic est dicendum quando quis potest se licite commendare et quando est veniale et quando est mortale 153, 109vb Hic contra morem vini gerit unda saporem 308, 228va Hic iam est pars executiva in qua prosequitur intentum volens depellere ignorantiam depositionis circa quid sciendum quod duplex est ignorantia scilicet ignorantia abnegationis et ignorantia depositionis 167, 1rb Hic possit quaeri de tempore et loco et sic de aliis. Ad illud respondet evangelista Iohannes 2 53, 85rb Hic post postillationem evangelii alias habitam dicam solum de illa parte Venite ad me 53, 119vb Hic s. Hieronymus sacerdos fuit ordine peregrinationis … Frater Iacobus de varietatibus legendarum 165, 73va Hic sumat peccatrix anima confidentiam tripliciter. Quia Christus venit quaesivit et salvum fecit venit nascendo quaesivit praedicando salvum fecit moriendo 162, 7rb Hic tria possunt notari scilicet Mariae assumptio cum dicitur Extendit Noe manum secundo festivitas venerabilis Mariae cum dicitur Apprehensam columbam tertio assumptionis securitas cum dicitur In arcam 617, 182v His verbis possumus quattuor notare. Primo quis sit Iacob secundo quod dormivit tertio quae sit scala quarto quod angelos vidit 162, 3va Historia haec est de David quomodo planxit filium suum Absolon. Historia nota est scilicet quid significat 617, 163v Historia passionis Christi fuit per istum modum sicut tunc dicit beatus Iohannes evangelista qui tunc passionem filii Dei scripsit 174, 156Ira Hoc evangelium facit memoriam de Christi passione et secundum Wilhelmum Duranti 53, 152ra Hoc facite in meam commemorationem (1 Cor 11,24). In hodierna epistola dominus noster Iesus Christus de hoc mundo per passionem suam acerbissimam 53, 257vb – Sacramentum mei corporis sumite quo ad fideles et conficite quo ad presbyteros 53, 258rb Hoc opusculum dividitur in tres partes scilicet in exordium in quo benevolentia captatur tractatum 161, 9ra Hodie agimus festum omnium angelorum et hoc propter plures causas. Primo quia singulariter omnes honorare non possumus. Isto die omnes honoremus 78, 100va Hodie carissimi dies celebratur insignis et festiva in tanto maioris devotionis affectu a singulis Christi fidelibus est celebranda 174, 163ra Hodie divina dies resurrectionis colitur in quo (!) perfecta requies designatur 53, 322rb Hodie in ecclesia agitur de duabus personis scilicet de Iuda qui est de honore suo detrusus 78, 35ra Hodie sancta mater ecclesia duas processiones recolit 153, 79rb; 174, 162va Hodie sancta mater ecclesia incipit celebrare adventum domini et hanc observantiam recepit a sanctis patribus 53, 1ra Hodie triplex festivitas celebratur. Una est de adoratione magorum 617, 139r Hodiernae diei festivitas magna est. Unde merito dicitur apparitio 153, 23ra Hodiernae diei gaudiosa festivitas nos admonet et accendit aliquid de temporali Christi 53, 52rb Hodiernam paschalem videlicet festivitatem inter ceteras festivitates Christi praecipuam 53, 313vb Homo est optimum eorum quae sunt in mundo quattuor Ethicorum et mundus 155, 123r Hora est iam nos de somno surgere (Rm 13,11). Egregius praedicator et apostolus invitans nos ad opera virtutum. In istis verbis tria facit. Primo allegat temporis opportunitatem 174, 168ra Hos vocat in primis Pelagi salvator 308, 228va Iam de mari composita fuerunt multa super primum librum. Sed adhuc super secundum quaeritur de perpetuitate vel generatione maris scilicet utrum fuerit perpetuum 440, 319ra Ibi dic historiam qualiter Mardochaeus somniavit duos dracones 617, 147r Iericho interpretatur luna et signat mundum propter suum defectum 162, 2va Iesu mi dulcissime felicissima sapientia verbum patris principium et finis alpha et o omnium entium 353, 14v Igitur locuturus hodie nunc amplius de dira Christi morte et innocenti passione 53, 281ra Illo nos igne spiritus sanctus inflammet quem dominus noster Iesus Christus 353, 36r Illud digne commendatur quod in se consideratum bonum apparet et aliis comparatum excellit 31, 84va Illud est prius quo non existente non sunt alia sed sine fide 153, 67va 438 Illumina quaesumus domine – In praesenti enim evangelio Illumina quaesumus domine populum tuum et splendore passionum charismatum et gloriae rationalis ac beatissimae animae unigeniti filii tui domini nostri Iesu Christi 353, 188v Immensa divinae largitatis beneficia exhibita populo Christiano inaestimabilem ei conferunt dignitatem 353, 56v Imminente hora qua Christus dominus voluit pro nobis crucis passionem et mortem pati 53, 328va Imperatoris susceptio est triumphi imitatio. Triumphus namque regis exhibebatur 483, 72v Imperavit febri et dimisit illam (Lc 4,39). Pro huius thematis introductione invenio tres scholas videlicet Origenis, Augustini, Hieronymi. Prima schola est Origenis cuius talis fuit opinio seu doctrina 597, 30rb In annuntiatione dominica Deus homo factus est et die eodem passus, Ioseph venditus, Isaac immolatus, Adam formatus 52, VDS In conclusione prima dicitur quod de virtutibus potest esse scientia arguitur virtus non est scientialis 440, 91rb In exordio praesentis negotii Deo scientiarum domino humiliter invocato protestamur quod circa puncta nuper in forma publicae scripturae de oppido Ulmensi Constantiensis dioecesis transmissa 165, 129r In fontibus ecclesiarum non in privatis domibus debet baptisma concedi 52, 39ra In his verbis agitur de duobus lignis. Primo de ligno salvationis in quo homo fuit vivificatus 617, 187Ir In his verbis notandum est quod sex sunt quae invitant nos ad petendum 617, 169r In his verbis notatur magna humilitas Christi qui servum centurionis dignatus est visitare 153, 39rb In his verbis tria videnda sunt scilicet quae sit dilecta quis sit dilectus quo et quomodo abiit. Circa primum sciendum quod dilecta est beata virgo 617, 170r In historia ecclesiastica habetur de institutione vitae sanctorum patrum in Aegypto 165, 111rb In hoc differunt Graeci a Latinis quia Graeci similiter omnes dicunt 52, 1ra In hoc evangelio considerantur triplices personae scilicet Christus discipuli et turbae 53, 242vb In hoc verbo Gabriel angelus sacramenti regalis conscius ac Dei fore virginis paranymphus describitur 31, 49vb In hodierna epistola dominus noster Iesus Christus de hoc mundo per passionem suam acerbissimam 53, 257vb In illo die quo videtur corpus Christi victus necessarius retribuitur lenia colloquia dimittantur 167, 71va In illo evangelio duo notantur scilicet dignitas vocantis cum dicitur Sequere me 617, 187Iv In illo tempore dixit Iesus discipulis suis. Scitis quia post biduum pascha fiet et filius hominis traditur ut crucifigatur … (Mt 26,2). Quia nullus evangelistarum materiam dominicae passionis plene ac complete conscripsit 756, 1ra In illo tempore respondens Iesus dixit Confiteor tibi pater domine caeli et terrae quia abscondisti haec a sapientibus et prudentibus etc. (Mt 11,25). Dominus Deus non invitat solum cottidie per suos nuntios praedicatores sed etiam per se ipsum 53, 119va – Hic post postillationem evangelii alias habitam dicam solum de illa parte Venite ad me 53, 119vb – Sicut inter virtutes morales quae pertinentes sunt ad mores exteriores praecipue praecipua est gratitudo 53, 156rb – Quia Christus multa signa fecit in civitatibus Iudaeorum ipsi nihilominus in sua pertinacia permanserunt 53, 156vb In illo tempore stabat Maria ad monumentum foris plorans et reliqua (Io 20,11). Homilia Origenis. De praesenti sollemnitate locuturus auribus vestrae caritatis dilectissimi amor venit ad memoriam 161, 6ra In istis verbis describit salvator adventum suum ad iudicium extremum 53, 20ra In latinis quidem libris nullum auctorem invenire potui qui de urina certam et authenticam cognitionem dederit 748, 138ra In matutinis domine precibus pater sancte famulos tuos exaudi 405, 77r In mentibus nostris quaesumus domine tuae gratiae lumen infunde ut qui conceptum et natum de virgine Deum verum et hominem confitemur 353, 174r In morbo materiali sunt habendae istae intentiones. Prima administratio regiminis debiti 748, 52r In nomine domini Amen. Circa processum iudiciarium in causis delegatis et subdelegatis et ordinariis primo praemittitur cum impetrator rescripti 614, 150r, 151r – Quia in libris sacrorum Canonum et in legibus imperialibus plerumque restat difficultas vocabulorum 155, 18ra – Splendor paternae gloriae 76, 66r In nomine sanctae et individuae trinitatis et in honore sanctae Mariae perpetuae virginis et sanctae Margarethae virginis et martyris omniumque caelestium virtutum consecrata est haec ecclesia anno dominicae incarnationis millesimo centesimo quinquagesimo sexto 293, 22r In odium quoque Christianorum Iulianus templum 308, 149vb In omnibus doctrinis quae secundum ordinem currunt incessus sunt secundum rerum ordines. Una earum est quae fit secundum 453, 1v In peccato vestro moriemini (Io 8,21). Pro huius thematis introductione invenio tres scholas scilicet caeli, paradisi, mundi. Prima ergo schola fuit caeli. Nam in principio creavit Deus caelum et terram 597, 18vb In praecedenti sermone dixi de signis adventus diei iudicii 53, 13ra In praesenti enim evangelio instruit Maria parentes qualiter se debeant habere ad filios 153, 27va In praesenti evangelio – Iohannes qui non est pater meus 439 In praesenti evangelio dominus ostendit multorum auditorum Christi devotionem 53, 139ra In praesenti evangelio monet nos dominus quatenus praeparemus nos ad istam ultimam rationem quam oportet nos facere in extremo iudicio 53, 7ra In praesenti evangelio ponuntur duo principalia. Primum de Christi passione 53, 146rb In quadam legenda transmissa de Roma 125, 199vb In quibus verbis monemur ne a bono iam incepto desistamus 153, 94va In sempiternis nihil violentum. Qui ergo alte in caelo vult 153, 46vb In summis festis ad missam dicitur una. Tantum collecta nisi sit coniuncta diei. In defunctorum collectam dicere missa de sanctis poteris sed illa paenultima fiat 167, 1rb In verbis istis apostolus duo facit, ostendit enim fidelis animae sanctitatem et exsecratur sanctitatis violatorem 162, 8ra In verbis istis notandum quod ad impetrandum salutaria quattuor oportet considerare videlicet a quo sit petendum quid sit petendum qualiter sit petendum quando sit petendum 617, 168v In verbis istis ostendit nobis spiritus sanctus quaedam signa in beato Nicolao propter quae signa obtinuit benedictionem omnium gentium 78, 4rb In verbis istis quattuor notantur. Primum est quod dominus dicit Ecce ego mitto angelum meum etc. Secundum est quod dicit 78, 29rb In verbis istis tria notantur de sancto Bernhardo. Primo sanctitas vitae cum dicitur Vidi alterum angelum descendentem de caelo amictum nube. Primus enim angelus fuit beatus Benedictus 78, 80ra In verbis propositis notatur triplex martyrium quod suscipit beatus Bartholomaeus fuit enim fustigatus deinde vivus excoriatus postremum decollatus 617, 183r Incipit tractatus venerabilis doctoris Iohannis Gerson olim cancellarii Parisiensis de oratione et suo valore 754, 1r Inclina domine aurem tuam ad preces nostras quibus misericordiam tuam suppliciter deprecamur ut animam famuli tui 654, 86v Inclinato capite emisit spiritum … (Io 19,30). Iohannes de Sancto Geminiano dicit Mors salvatoris carissimi quam hodie recolimus fuit ultima consummatio dominicae passionis 153, 86ra Infer digitum tuum huc (Io 20,27). Sciendum primo quod Christus servavit cicatrices vulnerum in suo corpore glorificato 153, 104va Infirmitatem paupertatem tribulationem atque omnem angustiam nostram 654, 11v Infunde quaesumus domine Deus tuae cordibus lumen sapientiae 353, 22v Ingressus Iesus ambulabat Iericho (Lc 19,1). Iericho interpretatur luna et signat mundum propter suum defectum 162, 2va Iniquitates nostrae contenderunt contra nos domine libera nos 654, 18v Inspice et fac secundum exemplar quod tibi in monte monstratum est (Ex 25,40). Nota quod Deus duo genera sanctorum habet in caelo quosdam abscondit a nobis et quosdam nobis monstravit 617, 191v Intentiones habendae in febribus excerptae ex mente Avicennae ex locis universalibus et particularibus. In cholerica interpolata pura sunt intentiones. Est administratio regiminis debiti sex rerum 748, 41r Intentionis discussio – ne propter vanam gloriam 167, 72r Inter cetera quae reconciliant peccatorem Deo postquam in peccatum mortale corruit est recognitio 53, 236vb Inter doctores populi sedet et seniores 308, 228va Inter medicinas et corpora currunt proportiones quae non comprehenduntur nisi cum experimento 748, 52v Interrogatus Socrates a quodam quomodo posset optime respondere seu dicere. Respondit Si nihil 123, 1r Interveniat pro nobis domine Iesu Christe nunc et in hora nostrae mortis apud tuam clementiam gloriosa Dei genitrix virgo Maria 353, 163r Interveniat pro nobis quaesumus Deus sanctus tuus Lucas evangelista qui crucis mortificationem 405, 219v Interveniat quaesumus domine Iesu Christe pro nobis apud tuam ineffabilem clementiam nunc et in hora mortis nostrae 353, 114r Iohannes abbas cum quendam qui per quadraginta annos in eremo habitaverat interrogasset 308, 48vb Iohannes de Sancto Geminiano dicit Mors salvatoris carissimi quam hodie recolimus fuit ultima consummatio dominicae passionis 153, 86ra Iohannes de Sancto Geminiano dicit quod totum officium praesentis diei dominicae continet quasi quaedam nova et laeta 153, 71vb Iohannes de Sancto Geminiano Videmus naturaliter quod lux habet expellere tenebras. Christus autem est lux 153, 69ra Iohannes episcopus servus servorum Dei. Ad perpetuam rei memoriam valitura canonum. Non est dubium pertinere cum statuta a se vel praedecessoribus suis edita obesse percipit potius quam prodesse ne ulterius adesse valeant providere. Dudum si quidem felicis recordationis Nicolaus papa IIIus praedecessor noster attente prospiciens 99, 33vb Iohannes qui non est pater meus nec consanguineus duxit matrem meam in uxorem suam. Ipsa moritur postmodum 155, 10ra 440 Iordanis sacri subit – Magnam hodie et praecipuam Iordanis sacri subit hic baptisma lavatri 308, 228va Ipsi soli servies (Mt 4,10). Pro huius thematis introductione invenio tres scholas scilicet Eremitarum, Praedicatorum, Minorum. Prima schola est Eremitarum in Aegidio qui videtur velle tenere quod si primus homo non peccasset 597, 7ra Ista dies praecipue maior est omnibus festivitatibus 174, 155ra Ista scribit beatus Lucas et sunt verba nostri salvatoris suam dilectionem quam ad quendam humanum habuit 162, 1ra Iste est liber sextus Physicorum, in quo philosophus determinat de divisione motus et quietis in suas partes 163, 1r Iste liber Physiologus intitulatur et dicitur a physis id est natura 155, 151r Isti sunt filii […] Iohannes bonus et Lafrancus filius Ambrosii est Albertus Zicerius 360, 26r Ita hortatur nos praedicator paenitentiae beatus Iohannes Baptista in deserto praedicans baptismum paenitentiae Matthiae 3 et Lucae 3 quod specialiter nobis dicitur hoc sacratissimo ieiuniorum tempore legimus 53, 227v Item cum quidam frater in subscripta manens 125, 199rb Item dum exercitus domini regis Roberti 125, 199ra Item Favinus de Slappis 125, 199rb Item Iohannes infans cum sorore maiore 125, 199va Item magister barbatus sancti Georgii 125, 199ra Item maiora de terra 125, 199va Item miles quidam Gallicus familiaris 125, 199rb Item mundus assimilatur duobus viris scilicet patri et filio quorum unus scilicet pater equitabat super asinum et filius pedester ambulabat 153, 161ra Item nota quod febricitantibus acutarum febrium et etiam aliis qui patiuntur 748, 212v Item nota quod per quidditatem animae intellegitur summa et essentia ipsius animae 163, 79v Item Petrus frater domini Matthaei de Salerno 125, 199rb Item pro secundo sermone in hac die tria sunt dicenda. Primo quomodo quattuor genera hominum 153, 84ra Item quaedam mulier in Arriano peperit 125, 199rb Item quidam nobilis falconem unum perdiderat 125, 199rb Item si alicui presbytero praesentaretur femella et ipse putaret masculum 550, 4vb Item si est haereticus. Item si publicus peccator poena usurarius concubinarius etc. 145, 314va Item ubi quis in testamento disposuit 145, 314vb Iuste iudicate filii hominum et nolite iudicare secundum faciem sed secundum iustum iudicium iudicate ut ostendatis vero diligere iustitiam qui iudicatis 107, 1ra Lacrimis rigavit pedes meos (Lc 7,44). Pro huius thematis introductione invenio tres scholas scilicet arborum et plantarum, vaporum et nebularum, dolentium animarum. Prima schola est plantarum et arborum in qua talis doctrina legitur quod quando arbores plorant 597, 37rb Laetifica et adiuva qui sumus omnipotens et misericors Deus dominicae annuntiationis et conceptionis gaudia recolentes 353, 172r Legitur de quinque virgis laudabilibus per quas beata virgo Maria merito figuratur 617, 146v Legitur de sancto Severino quod vivens mortuos suscitavit imo omnem infirmitatem 308, 197vb Legitur de sancto Udalrico cum vellet causa devotionis 308, 200va Legitur quod in dioecesi Tridentina quidam latro fuerat famosus 308, 193ra Liber Aviani poetae in quo primo introducit fabulam de villana et lupo ostendens in eo quod fraudulenti mulieri non est fides 155, 155v Liber ergo iste in quinque partes dividitur. Prima est de essentia febris eiusque definitione febris naturam et causas 748, 162ra Licet laico plus in die 483, 28r Licet multae sunt causae in particulari quare lites et turbationes surgunt inter homines in mundo 53, 325ra Licet mundus iste primo appareat valde amicus tranquillus et pacificus hominibus 53, 114va Licet secundum Augustinum in quodam sermone ad fratres Non debeamus loqui sublimia 53, 242rb Loquendo de manducatione huius venerabilis sacramenti sciendum quod hoc sacramentum tripliciter sumitur 153, 158va Magister Iacobus scutellarius commendabilis vitae 125, 198vb Magistri scholarum litterarum aiunt animam esse substantiam simplicem speciem naturalem distantem a materia corporis sui organum membrorum 597, 57vb Magnam hodie et praecipuam carissimi omnique devotione recolendam agimus sollemnitatem ad cuius celebrationem venerandam venerabilis Hugo de Sancto Victore in quodam sermone ipsius sollemnitatis nos devote invitat 174, 166va Maiestatem tuam domine – Monachus quidam vitae laudabilis 441 Maiestatem tuam domine supplices exoramus ut sicut iugiter sanctorum tuorum 405, 221v Maiestatem tuam domine suppliciter exoramus ut sicut ecclesiae tuae beatus Andreas 405, 219v Mane nobiscum domine (Lc 24,29). Circa quod sciendum quod sunt sex quae debent hominem retrahere ab omni peccato mortali 153, 96ra – Haec verba canit ecclesia per illud sacrum tempus paschale usque ad ascensionem 153, 91vb – In quibus verbis monemur ne a bono iam incepto desistamus 153, 94va Manus Zorobabel fundaverunt eam (Za 4,9) scilicet animam per istum Zorobabel eam 165, 144rb Marcus Valerius Probus unicae vetustatis amator 780, 1r Maria du hochgelobter Namen ein Khönigin des Himels 397, Iv Maria Magdalena et Maria Iacobi et Salome emerunt aromata ut venientes ungerent Iesum (Mc 16,1). Divina scriptura istam habet proprietatem quod in una sui parte fideles admonet lacrimari 53, 304vb – Hae duae fuerunt ultimae sorores ex parte matris cum virgine gloriosa 53, 299vb – Praesens Novi Testamenti praecipua festivitas inter ceteras venerabilis toto orbe terrarum 53, 299ra Maria mater gratiae mater misericordiae tu nos ab hoste protege in hora mortis suscipe 405, 208v Martha ministrabat (Io 12,2). Pro huius thematis introductione invenio tres scholas scilicet internas, alternas, subalternas. Prima est interna et est voluntatis et in hac legitur quod ea quae sunt in homine ipsi voluntati praesenti principaliori et digniori subalternas serviunt 597, 42ra Matutina supplicium vota quaesumus domine propitius intuere 405, 31r Medicina est scientia sanorum aegrorum et neutrorum 453, 1v Memento ut sanctifices diem sabbati. Per sabbatum intellegimus sollemnitates sanctorum in quibus quam plurimum homines vacant commessationibus et ebrietatibus 308, 225va Memoriam esse thesaurum inventorum eleganter ait Cicero in Rhetoricis ad Herennium libro tertio 4, 1ra Mentes nostras quaesumus domine spiritus sanctus reparet 353, 34r Miles quidam apud Fridericum imperatorem in multis est accusatus 308, 191rb Miles quidam beatum Thomam apostolum ardenter diligebat 308, 196rb Miles quidam in dandis decimis suis tantae erat iustitiae ut ei summum esset studium 308, 197ra Minos Iovis et Europae filius Androgeum Ariadnam Phaedram Deucalionem et Molum progenuit. Ex Deucalione Idomeneus ex Molo Meriones seu Mirion et Idomeneus alter nascitur. Item Tantalus Iovis filius genuit Pelopem 691, 1r Miseremini mei etc. (Iob 19,21). Notandum monere paenitentiae portando debemus fratres nostros exemplo Christi qui poenam peccatorum portavit 78, 113rb Miserere domine per sanctam annuntiationem adventum nativitatem ieiunium 654, 91v Miserere mei fili David (Mt 15,22). Pro huius thematis introductione tres scholas reperio scilicet illustrium magnificorum secunda eloquentium tertia decipientium et proditorum. Prima schola fuit illustrium etc. 597, 12ra Miserere nostri Deus omnium et respice nos (Sir 36,1). Pro huius thematis introductione tres scholas reperio ethicam seu monasticam, oeconomicam seu familiarem, politicam seu civilem. Prima schola est monastica in qua legitur 597, 14rb Misericors pater qui est prope est omnibus invocantibus eum in veritate quia nos orphanos non relinquit 53, 344rb Miserunt Iudaei a Hierosolymis sacerdotes et levitas ad Iohannem ut interrogarent eum Tu quis es Iohannis 1 capitulo (19). Et in evangelio hodierno sicut scriptura sacra Veteris et Novi Testamenti persuadet fideles ut diligant et amplexentur humilitatem 53, 30ra – Secundum Originem Iudaei videntes prope esse tempus adventus Messiae 53, 30va Misit digitos suos … (Mc 7,33). Digitos in aures surde (!) et tandem rogaverunt tam ut imponeret sibi manum 177, 134rb Missae sunt celebrandae in locis Deo consecratis vel in locis in quibus episcopus permiserit celebrare quia melius est missam non celebrare aut non audire 155, 14rb Missus est Gabriel angelus a Deo (Lc 1,26). In hoc verbo Gabriel angelus sacramenti regalis conscius ac Dei fore virginis paranymphus describitur 31, 49vb – Vehementia desiderii, fervore devotionis, puritate orationis fontem attingit pietatis 171, 88rb Mittens Herodes occidit omnes pueros … (Mt 2,16). Ex quo hodie peragitur festum illorum puerorum innocentium qui ab iniquo Herode interfecti sunt 153, 14ra Mittite partes eis qui non praeparaverunt sibi (2 Esr 8,10). Consuetudo in mundo est quando aliquis usque in mortem in carcere cruciatur mittit ad amicos suos de quibus maxime confidit 78, 112ra Modicum et iam non videbitis me (Io 16,16). Haec verba locutus est dominus Deus et ante passionem suam post cenam 153, 109vb – Imminente hora qua Christus dominus voluit pro nobis crucis passionem et mortem pati 53, 328va Modus est nobilium et aliorum honestorum virorum quod ad filiorum suorum custodiam et instructionem ponunt suos servos 78, 98va Monachus quidam vitae laudabilis sed corpore debilis fuit 308, 192vb 442 Mortem non gustabit – Non sumas nomen Mortem non gustabit in aeternum (Io 8,52). Pro huius thematis introductione invenio tres scholas scilicet planetae, cometae, monetae. Prima est planetae et in hac legitur vita secundum quod melior est constellatio vel planeta de tanto efficimur levioris vitae 597, 35vb Mortuus est dives et sepultus est in inferno (Lc 16,22). Pro huius thematis introductione invenio tres scholas scilicet Sardanapalli in Asia, Catilinae in Roma et Attilae in Hungaria. Prima ergo schola est Sardanapalli 597, 22va Mos paganorum versus est in religionem Christianorum 483, 76r Motus magnus factus est … (Mt 8,24; Mc 4,37; Lc 8,23). Ubi sciendum est quod per mare intellegitur praesens mundus et per motum magnum 153, 41va Mulier dum parit tristitiam habet (Io 16,21). Ex quo in verbis praemissis fit mentio de partu mulierum. Tunc in praesenti sermone tria sunt dicenda 153, 112vb Mulier magna est fides tua … (Mt 15,28; Mc 7,25) etc. Aristoteles 8o Physicorum Illud est prius quo non existente non sunt alia sed sine fide 153, 67va Mulier uxor alicuius confitetur sacerdoti se tenuisse filium proprium coram episcopo 155, 9vb Multae fuit potestatis erga daemones pellendos non solum ex hominibus 308, 28rb Multae subtilitatis fuit erga daemones cognoscendos. Quadam vice diabolus in forma regis purpura 308, 28rb Mundat leprosum tangens sub lege perosum 308, 228va Mundus autem gaudebit (Io 16,20). Hic est dicendum quare gaudium mundi est fugiendum et quando est mortale 153, 115va Musica humanae vitae necessaria nedum apud Graecos aut Romanos verum etc. apud barbaros semper magna laude prosecuta est 776, 13r Nabuchodonosor quippe rex Babylonis bellica manu Ierusalem destruxit 483, 38v Nec ego te condemnabo (Io 8,11). Pro huius thematis introductione invenio tres scholas scilicet vulgaricam, canonicam, theologicam et in his legitur quod nullus possit iudicari 597, 32ra Nemo potest conficere corpus Christi nisi sacerdos qui rite fuerit ordinatus secundum claves ecclesiae 155, 10rb Nemo potest venire ad me nisi pater meus traxerit eum (Io 6,44). Sciendum quod circa praedicta verba tria sunt dicenda. Primo quomodo Deus trahit et vocat hominem 153, 144rb Nemo propheta acceptus est in patria sua (Lc 4,24). Pro huius thematis introductione tres scholas invenio scilicet filiorum, inimicorum, discipulorum. Prima schola est filiorum. In hac considero inter filios primi parentis 597, 29ra Nescitis quid petatis (Mt 20,22). Pro huius thematis introductione tres scholas reperio videlicet mundi, carnis, daemonis. Prima schola est mundi in prima legitur apparentia sive existentia 597, 21rb Nicolaus Iohannis marescalci 125, 199va Nimis honorati sunt amici tui Deus (Ps 138,17). Dicitur vulgariter Bonum ferrum trahit homini argentum de bursa 617, 188v Nisi quis renatus fuerit ex aqua … (Io 3,5). Sciendum quod triplex est nativitas naturalis spiritualis et caelestis 153, 150va – Ex praemissis patet quod quam cito homo moritur qui adhuc stat in innocentia baptismali 153, 152va Noctis sub silentio tempore brumali deditus 308, 189ra Nomen virginis Maria (Lc 1,27). De dulcissimo nomine ipsius Mariae adhuc amplius videamus. Est notandum quod nomen huius dulcissimae virginis est a Deo revelatum 31, 58rb – De nomine huius ineffabilis virginis adhuc est plus dicendum. Et est notandum quod haec virgo postquam Christum concepit reperitur in scriptura 31, 60va – Nulla res potest amari nec laudari nisi prius cognoscetur dicente Augustinus (!) de trinitate 31, 55va – Prius dictum est de nomine. Nunc dicendum est de hoc vocabulo Maria. Et est notandum quod quattuor genera volatilium solent nidos suos ornare 31, 65va Non enim volebat in Iudaeam ambulare quia quaerebant eum Iudaei interficere (Io 7,1). Pro huius thematis introductione tres scholas invenio scilicet Martis, partis, mortis. Prima est Martis. Nam Mars ut pluries dictum est planeta malevolus est et nati sub illo sunt pessimi 597, 36va Non est vir in domo sua abiit via longissima secum tulit sacculum pecuniae in die plenae lunae reversurus (Prv 7,19). Verba proposita possunt esse interioris hominis id est animae respondentis ad quaestionem sibi factam 617, 171r Non rapinam arbitratus est esse se aequalem Deo (Phil 2,6). Pro huius thematis introductione invenio tres scholas angelicae naturae, humanae creaturae, artificialis figurae et in omnibus his legitur forma rapinae. Prima schola est angelicae naturae in qua legitur rapina 597, 40va Non solum Christi verba sunt nobis in documentum sed etiam Christi facta sunt nobis in exemplum 153, 30va Non sumas nomen Dei tui invanum. Augustinus: Nomen Dei sumit invanum qui semper turpia verba profert ex corde. Unde legitur in dialogo Gregorii 308, 211rb Non sumus filii – Nuptiae factae sunt 443 Non sumus filii ancillae sed liberae (Gal 4,31). Pro huius thematis introductione tres scholas invenio scilicet potentiarum, naturarum, grammaticorum. Nam prima est potentiarum et quia habemus plures potentias 597, 32rb Nonae Aprilis norunt quinos 360, 26r Nos cum prole pia benedicat virgo Maria 405, 209v Nos enim videmus ad sensum quod quando aliqui evaserunt mortem qui fuerunt in bello fortissimo ubi multi mortui fuerunt ad suos dilectos veniunt et dicunt 174, 90vb Nota in praecedenti evangelio dominicali dicitur quod publicani et peccatores 174, 205va Nota Octavianus imperator dictus est Augustus primus quia regnum Romanorum notabiliter auxit 53, 43va Nota oleum factum per decoctionem medicinarum 748, 54ra Nota praedicatores dicuntur pastores quibus dicuntur Iohannes XXIo Afferte mihi de piscibus 174, 222ra Nota quando tritulator est debilis et habet grave flagellum in manu 174, 187rb Nota quod Deus duo genera sanctorum habet in caelo quosdam abscondit a nobis et quosdam nobis monstravit 617, 191v Nota quod est triplex via. Prima in mundum, secunda per mundum, tertia extra mundum. Prima via in mundo (!) est lamentabilis 78, 64vb Nota quod inter omnia festa laetabunda totius anni hoc festum nativitas est tanto laetius 153, 1va Nota quod omnis dispositio 532, 124v Nota quod post symbolum apostolorum quod dicitur Credo in Deum concilia praedicta fuerunt olim a sanctis patribus de summa trinitate et fide catholica contra quosdam haereticos de quibus quattuor conciliis legitur 155, 106ara Nota quod quidam Deum recipiunt in domum suam et non in animam quidam in animam et non in domum quidam in animam et in domum quidam nec in animam nec in domum 162, 1va Nota quod sponsa Christo debet se praeparare ad eius adventum quinque modis. Primo se lavando 162, 6va Nota quod succus paeoniae vulneribus noviter fissis applicatus nondum ad sanationem provenientibus curat 748, 69v Nota rupto apostemate et extracta materia 748, 66r Nota secundum Averroim quod medicina est de subiecto cuius subiecti ratio in medicina salvatur 453, 1v Nota verum secretum in provocatione urinae. Recipe corticis interioris 748, 54ra Notandum circa textum nonnulli priorum opinati sunt quod quando anima ingrediebatur corpus 163, 35r Notandum monere paenitentiae portando debemus fratres nostros exemplo Christi qui poenam peccatorum portavit 78, 113rb Notandum quod beatus Thomas Christum discipulis apparentibus non vidit quia non fuit cum eis sed post octo dies eum vidit 153, 103va Notandum quod Octavianus imperator 42 anno imperii sui quia 57 annis regnavit et versus finem valde pacifice 53, 52vb Notandum quod quintuplex est anima scilicet anima immunda anima rapta anima sepulta anima salvata anima cruciata 617, 190r Notandum quod sicut sunt septem utilitates sive efficaciae quas gratia confert animae de quibus dictum est in sermone praecedenti dicendum est 31, 71va Notandum tamen etc. quod quidam sacerdos 550, 24vb Nova lux oriri visa est honor et tripudium apud omnes populos urbes et provincias (Est 8,16). Quia nativitatem beatae Mariae agimus ideo gaudere debemus quia ipsa est stella 78, 91rb Nova veniens e caelo nuptiali thalamo (vgl. AH 51, Nr. 102, Str. 2). Nota quod sponsa Christo debet se praeparare ad eius adventum quinque modis. Primo se lavando 162, 6va Novicius indutus quantocius potest in generali confessione se expediat in qua non sit nimis scrupulosus 617, 95v Novissime audistis quomodo in hoc sacratissimo tempore matamur (!) ad paenitentiam 53, 232va Nulla obstetrix nulla muliercularum sedulitas intercessit ipsa pannis involvit infantem 52, 38vb Nulla res potest amari nec laudari nisi prius cognoscetur dicente Augustinus (!) de trinitate 31, 55va Nullum etiam sacramentum est isto salubrius quo purgantur peccata 353, 58v Numquid Iudaeus litigans cum Christiano iurabit de calumnia super evangelio videtur 155, 6va Nunc ergo de confessione videndum est ad quam homo obligatur qui peccavit mortaliter ex Dei institutione et praecepto (Io 20,22) ubi confessionem instituit quando dixit apostolis Accipite spiritum sanctum 53, 205ra Nunc igitur de gula videndum est isto tempore sacro quia secundum Gregorium 53, 191ra Nunc igitur iuxta praemissa sicut dixi ad contemplationem 53, 252ra Nuptiae factae sunt in Chana Galilaeae (Io 2,1). De prima parte evangelii dicendum est sed referendo ad nuptias corporales de quibus loquitur specialiter evangelium 53, 89vb – Ex quo evangelium hodiernum mentionem facit de matrimonio et de nuptiis in quibus Christus Deus et homo cum benedicta matre 153, 35va – Hic possit quaeri de tempore et loco et sic de aliis. Ad illud respondet evangelista Iohannes 53, 85rb 444 O admiranda – Omnipotens et sempiterne Deus – Prima pars praesentis evangelii congruit multum praesenti tempore in quo celebrantur nuptiae et matrimonia 53, 92vb – Propter multiplicationem humani generis Deus specialiter instituit matrimonium 53, 84vb – Quamvis haeretici miseri mentiantur carnale martyrium esse illicitum eorum tamen falsitas manifeste apparet 153, 32va – Quamvis in nuptiis homines soleant convivia tenere haec autem caute tenenda sunt in Dei timore 53, 92rb O admiranda et laudanda tuae dispensationis gratia omnipotentissime domine Deus. Quia enim in natura tua ab homine videri non poteras ut hominem notitiae tuae participem faceres opus fecisti 397, 1r O altitudo divitiarum sapientiae et scientiae Dei … (Rm 11,33 und 36). Sciendum quod hodie festum trinitatis institutum est ut omnes fideles agnoscant unam simplicem deitatem et trinitatem 153, 147va O beata et gloriosa virgo Maria quis digne tibi valeat iura gratiarum 405, 209v O dies aeterne numquam deficiens et sol numquam occidens qui circa solis occasum sepultus suscepisti 405, 215v O domine Iesu Christe fili Dei vivi benedicti qui pro nobis peccatoribus agonizans in cruce praedilectam matrem 353, 177r O domine Iesu Christe aeterna dulcedo te amantium iubilus excedens omne gaudium et omne desiderium 654, 100r O Maria dominus tecum te associans tecum te custodiens tecum te glorificans 31, 82ra O quam magnum donum et mirificum bonum de Dei misericordia quia tanta est dignitas 167, 71va O refugium meum o liberator meus per immensum amorem qui te in patibulo crucis amarissimam mortem pro me subire coegit 353, 15v O regina virginum fac tecum gratulari / Quos hic in exilio vides tribulari 166, 52ra O stulti et tardi corde ad credendum (Lc 24,25). Sciendum quod secundum dictum Salomonis stultorum infinitus est numerus 153, 99rb O virgo virginum beatissima virgo Maria propter angustias et cruciam inaque cor tuum sustinuit 353, 81v O virgo virginum castissima Dei genitrix Maria propter planctum acerbi eiulatus 353, 89v O virgo virginum clementissima mater Maria propter doloris gladium qui pertransivit animam tuam 353, 84r O virgo virginum gloriosissima mater Maria propter gemitus et suspiria indicibiliaque lamenta quibus afficiebaris 353, 92v O virgo virginum praedulcissima mater Maria quando vidisti filium tuum 353, 79v O virgo virginum sanctissima Dei genitrix Maria propter gravamen et tormentum quo torquebatur 353, 77v O virgo virginum sanctissima regina propter gravamen et tormentum quo torquebatur spiritus tuus iuxta crucem 353, 86r O vos electi sumus namque fragiles et nullius veritatis 654, 4r O vos omnes qui transitis … Trenorum primo (Lam 1,12). Unde Iohannes de Sancto Geminiano dicit Conveniens est ut sentiat membrum quod sentit caput 153, 84ra Omne quod movetur necesse. Iste est septimus liber Physicorum in quo philosophus determinat de comparatione motuum 163, 10r Omne regnum in se divisum desolabitur (Lc 11,17). Pro huius thematis introductione invenio tres scholas scilicet Thebanam, Troianam, Romanam. Prima schola fuit Thebana et in hac pro sui tempore lecta fuit divisio 597, 28ra Omnes sancti tui quaesumus domine nos ubique laetificent et adiuvent ut dum eorum merita recolimus patrocinia sentiamus 353, 31r Omnes sunt administratorii spiritus in ministerium missi propter eos qui hereditatem capiunt salutis gloria id est hominis quibus promittitur vita aeterna (Hbr 1,14). Hodie agimus festum omnium angelorum et hoc propter plures causas. Primo quia singulariter omnes honorare non possumus. Isto die omnes honoremus 78, 100va – Modus est nobilium et aliorum honestorum virorum quod ad filiorum suorum custodiam et instructionem ponunt suos servos 78, 98va Omnia per ipsum facta sunt et sine ipso factum est nihil. Haec verba scripta (Io 1,3) sic possunt exponi. Omnia scilicet entia a Deo alia. Facta sunt per ipsum id est per verbum Dei. Unde Augustinus sermonem dirigens ad Deum 532, 123r Omnia scilicet entia a Deo alia. Facta sunt per ipsum id est per verbum Dei. Unde Augustinus sermonem dirigens ad Deum 532, 123r Omnibus martyribus praefertur. Sic etiam de loco vel eventu Felicis 483, 72r Omnipotens Deus ex supereffluenti non ex operibus iustitiae quae fecimus 53, 66rb Omnipotens et misericors Deus qui electos spiritus caelestium gaudiorum 353, 7v Omnipotens et misericors Deus qui hominem ad imaginem tuam formasti et ei angelum corporis et animae custodem 353, 6v Omnipotens et misericors Deus qui omnem animam confitentem et paenitentem 405, 134r Omnipotens et sempiterne Deus cuius natura bonitas cuius voluntas potentia cuius opus 654, 20v Omnipotens mitissime Deus – Per proprium sanguinem 445 Omnipotens mitissime Deus qui sitienti populo fontem 405, 134v Omnipotens sempiterne Deus confitentibus tibi famulis tuis pro tua pietate relaxa peccata 654, 23r Omnipotens sempiterne Deus dirige actus nostros in beneplacito tuo 405, 224r; 654, 64v Omnipotens sempiterne Deus famulos tuos dextera potentiae tuae a cunctis protege periculis 405, 213v Omnipotens sempiterne Deus maestorum consolatio laborantium fortitudo 405, 125r, 134v Omnipotens sempiterne Deus ne nos miseros peccatores permittas propter peccata nostra perire 654, 33r Omnipotens sempiterne Deus qui angelum tuum antequam nasceretur homo et ego nascerer ad custodiendum 353, 11r Omnipotens sempiterne Deus qui caelestia simul et terrena moderaris 405, 133v Omnipotens sempiterne Deus qui dedisti famulis tuis in confessione verae fidei aeternae trinitatis gloriam agnoscere 353, 30r, 121r; 405, 216v, 221r Omnipotens sempiterne Deus qui ex abundantia pietatis tuae et merita supplicum excedis 353, 52r Omnipotens sempiterne Deus qui facis mirabilia magna solus 405, 133v Omnipotens sempiterne Deus qui in unitate permanes et in trinitate semper consistis 353, 124r Omnipotens sempiterne Deus qui nos omnium sanctorum tuorum merita sub una tribuisti celebritate 405, 218v Omnipotens sempiterne Deus qui vivorum dominaris simul et mortuorum omniumque 405, 135r Omnipotens sempiterne Deus respicere dignare super apostolicam dignitatem 405, 133r Omnis homo certis sciat ex causis et apertis / quod tres sunt sancti quos Ioseph nomine legi 293, 23r Omnis motus tendit ad aliquem terminum per debita media 153, 74va Omnis qui petit accipit etc. (Mt 7,8; Lc 11,10). In verbis istis notandum quod ad impetrandum salutaria quattuor oportet considerare videlicet a quo sit petendum quid sit petendum qualiter sit petendum quando sit petendum 617, 168v Omnis res per quascumque causas nascitur per easdem et dissolvitur (Decretum Gratiani, p. 2, C. 27, qu. 2, cap. 4). Ea facta quae dubia sunt ut quo animo in meliorem partem interpretemur 614, 282r Omnium magister et dominus Iesus Christus de mundo ascensurus ad patrem 550, 77ra Opera quae ego facio in nomine patris mei ipsa testimonium perhibent de me (Io 10,25). Pro huius thematis introductione invenio tres scholas scilicet veneficas, angelicas, truffaticas. Prima schola est angelica et in hac legitur magna opera facta fuisse et primo legitur de archangelo Michaele 597, 36vb Oportet ut extrahantur superfluctuentes humidae primo per flebotomiam 748, 71r Oremus pro omni gradu ecclesiae. Sacerdotes tui induantur iustitia 405, 132v Ostendat faciem suam nobis: et misereatur nostri 405, 89r Ostende nobis domine misericordiam tuam et salutare tuum da nobis 353, 41r Ostendite mihi numisma census (Mt 22,19). Nota praedicatores dicuntur pastores quibus dicetur Iohannes XXI Afferte mihi de piscibus 174, 222ra Paenitentiam agite … (Mt 3,2). Aristoteles 5o Physicorum Omnis motus tendit ad aliquem terminum per debita media 153, 74va Papa stupor mundi si dixero papa nocenti 327, 1r Partes orationis quot sunt? Octo. Quae? Devotio lacrimae dulcedo incendium ascensio meditatio contemplatio et languor. Devotio quid est 617, 2v Parvulus natus est nobis (Is 9,5). Circa nativitatem domini tria notantur. Primo natus describitur secundo nativitatis tempus tertio nativitatis utilitas. De primo sciendum quod fuit natus admirabilis ineffabilis formidabilis venerabilis 617, 136r Pastores loquebantur ad invicem Transeamus usque Betlehem et videamus hoc verbum quod factum est etc. (Lc 2,15). Hodiernae diei gaudiosa festivitas nos admonet et accendit aliquid de temporali Christi 53, 52rb – Notandum quod Octavianus imperator 42 anno imperii sui quia 57 annis regnavit et versus finem valde pacifice 53, 52vb Pateant aures misericordiae tuae domine precibus supplicantium et ut petentibus desiderata 353, 48r Pater clarifica nomen tuum (Io 12,28). Pro huius thematis introductione invenio tres scholas scilicet sapientiae, praesentiae, refulgentiae. Prima schola est sapientiae in qua legitur quod sapientium nomen clarificatur per famam in multis locis 597, 39va Per hoc convivium Veteris Testamenti figuratum est nobis convivium a Christo hodie institutum sacri corporis et sanguinis sui 617, 173v Per magnam misericordiam domini nostri Iesu Christi et per multitudinem miserationum suarum salvet et protegat 654, 17v Per proprium sanguinem introivit semel in sancta aeterna redemptione inventa (Hbr 9,12). Pro huius thematis introductione invenio tres scholas scilicet cordialem, pigmentalem, usualem. Prima est cordialis in qua legitur quod sanguis est summum elementum cordis 597, 35rb 446 Percusserunt me – Precibus et meritis Percusserunt me et vulneraverunt me custodes murorum (Ct 5,7). In verbis propositis notatur triplex martyrium quod suscipit beatus Bartholomaeus fuit enim fustigatus deinde vivus excoriatus postremum decollatus 617, 183r Perfecta Christi caritas confirmetur in nobis 405, 12v Perpetuum nobis domine tuae miserationis praesta subsidium 353, 10r Persaepe animadvertens hoc mortale hominum genus iniuriis casibus additum et infinita mortis 748, 90v Pestilentia est mutatio quaedam in aere facta propter admixtionem vaporum corruptorum vel sic est morbus quidem contagiosus 748, 72r Pestilentiae causa dicitur esse triplex. Nam vel est causa radix superior tantum et haec est superior et caelestis 748, 54v Petite et accipietis (Io 16,24). Benignitas salvatoris Dei nostri maior est in exaudiendo petitiones filiorum suorum 153, 119vb – Ex quo dominus noster Iesus Christus in verbis praemissis invitat nos ad orandum tunc hic quaeritur 153, 123rb Petite et accipietis (Lc 11,9). In his verbis notandum est quod sex sunt quae invitant nos ad petendum 617, 169r Petre amas me. Tu scis domine quia amo te (Io 21,17). Dicit beatus Iohannes in suo evangelio quod dominus interrogavit beatum Petrum de sua dilectione dicens 617, 175v Petrus apostolus dicit Paenitentiam agite et baptizetur 181, 41rb Phlebotomia electiva debet fieri parum post ortum solis et post primam et secundam digestionem et superfluitum expulsionem 748, 52v Pietate tua quaesumus domine nostrorum absolve vincula peccatorum 405, 132v Pii esse debemus in parentes in sanguine nobis iniunctos ad necessitatem praecipue autem post debemus esse in Deo 755, 10vb Piissimus salvator Iesus Christus totiusque consolator Deus omnis suae pietatis tantam humani generis imperitiam et ignaviam necnon infirmitatis 85, 1r Placeat tibi sancta trinitas hoc obsequium servitutis meae et praesta ut hoc sacrificium 252, 21v Plebani quia post praelatum maioribus quam ceteri fratres sunt periculis expositi 76, 63v Plebs ab imatu parit Herodis cruciatu 308, 228va Plures sunt agri in quibus seminat 177, 44vb Ponam arcum meum in nubibus caeli et erit signum foederis mei (Gn 9,13). Haec verba possunt exponi de beata Maria Magdalena quae intellegitur per arcum caelestem propter tria. Primo ratione materiae 78, 69vb Porrige nobis Deus dexteram tuam et per intercessionem sanctissimi Dei genitricis Mariae auxilium nobis supernae virtutis impende 353, 153Ir Post considerationem eorum quae pertinent ad mysteria verbi incarnati considerandum est de ecclesiae sacramentis 38, 144rb Post prooemium in quo breviter edocemur 253, 1ra Postquam consummati sunt dies octo ut circumcideretur puer vocatum est nomen eius Iesus etc. (Lc 2,21). Ex quo tota Christi vita et conversatio nostra est instructio 53, 63ra – Sciendum quod Christus die octavo circumcisus est et sanguinem suum fudit 153, 21ra – Sciendum quod Deus creavit primos parentes iustos iustitia naturali id est sine omni obliquitate 53, 63va – Solent homines in novo anno munuscula sibi invicem dare 153, 19rb Postquam locutus est eis Assumptus est in caelum … (Mc 16,19). Sciendum quod ab origine mundi non fuit tale festum in caelo nec umquam usque post iudicium erit 153, 126ra Praelatus monasterii qui tam verbo quam exemplo fratribus sibi commissis praeesse debet 76, 54r Praemissis in nomine Christi librorum tabulis sive capitulorum titulis sequuntur ex eis 52, 4ra Praesens evangelium mea non indiget expositione quia summa veritas 53, 134va Praesens evangelium tangit statum Christi tempore pueritiae circa 12 annum 53, 81rb Praesens Novi Testamenti praecipua festivitas inter ceteras venerabilis toto orbe terrarum 53, 299ra Praesta quaesumus omnipotens Deus ut familia tua per viam salutis incedat et beati Iohannis praecursoris 405, 217r Praesta quaesumus omnipotens Deus ut passionum charismatum et gloriae rationalis ac beatificae animae unigeniti filii tui domini nostri Iesu Christi 353, 189v Praesta quaesumus omnipotens Deus ut qui beati Pangratii martyris tui natalicia colimus 14820, 85v Praesta quaesumus omnipotens Deus ut qui gloriosos martyres Dionysium socios quam (!) eius fortes in sua confessione cognovimus 405, 219r Praetende domine fidelibus tuis dexteram caelestis auxilii 405, 207r Praetende quaesumus domine famulis et famulabus tuis 654, 65r Praeterea de lege illa vel errore quem cives tuos asseris statuisse videlicet ut si quis teneret per 30 annos 107, 79ra Praeterea quam divinum officium concernunt 76, 53v Precibus et meritis beatae Marie virginis liberet nos filius eius ab omnibus malis 353, 166r Precibus et meritis – Pro huius thematis introductione 447 Precibus et meritis beatissimae Mariae virginis et omnium sanctorum perducat nos dominus ad regna caelorum 353, 134r Precibus et meritis sanctissimae Dei genitricis virginis Mariae et omnium sanctorum suorum salvet et protegat 405, 14v Precibus et meritis sanctissimae suae matris salvat et protegat 353, 14r Presbyter attende capias haec dogmata mente / haec tibi norma datur per quam tua vita regatur 167, 71vb Presbyter potest supplere missam ab episcopo incohatam si episcopus non poterit propter infirmitatem 155, 13ra Prima missa est de exaltatione sanctae crucis … In secunda missa sic domine Iesu Christe qui iudicium innocentis sustinuisti … In tertia sic domine Iesu Christe qui innocenter et indigne in cruce es conclamatus … In quarta missa sic domine Iesu Christe per vulnera piissimi corporis et cordis tui … In quinta sic domine Iesu Christe rex caelorum per sepulturam tuam dignare animam 754, 26v Prima pars praesentis evangelii congruit multum praesenti tempore in quo celebrantur nuptiae et matrimonia 53, 92vb Prima pronosticorum finis laudabilis medicinae existit cum actor futura in singulis propendit quamvis omnes minime curare possit 453, VDS Primo debemus quaerere Christum cum plena cordis conversione 308, 201va Primo reprobatur superfluitas quia superflua sunt resecanda ab arte id est a iure canonico 155, 49vb Primum capitulum est quomodo confessor debet considerare conditiones confiteri volentis. Ubi notandum quod confessor sedens ad confessiones audiendas si confiteri volentem non cognoscit quaerat ab eo an sit suus parochianus 606, 1rb Primum dubium quare confert thyriaca et metridatum cum sint excellentis calefactionis et notum est quod quae augmentant caliditatem in corpore nostro 748, 64v Primus qui hic fuit Ninius idem fabricator fuit Thare ille dedit Abraham. Abraham dedit Ismaelitis 308, 228vb Prius dictum est de nomine. Nunc dicendum est de hoc vocabulo Maria. Et est notandum quod quattuor genera volatilium solent nidos suos ornare 31, 65va Prius dictum est quod gratia habet septem efficacias quas in anima facit et quod sunt septem praeparationes quibus homo ad gratiam se disponit 31, 73rb Prius dictum est quomodo Maria fuit gratiose salutata quod notabatur in hoc verbo Ave 31, 75vb Pro huius thematis introductione invenio tres scholas angelicae naturae, humanae creaturae, artificialis figurae et in omnibus his legitur forma rapinae. Prima schola est angelicae naturae in qua legitur rapina 597, 40va Pro huius thematis introductione invenio tres scholas Catonis, Tullii et Senecae. Prima schola est Catonis et quamquam intra illam bona vita legatur tamen inter alia qui ibi leguntur 597, 29vb Pro huius thematis introductione invenio tres scholas scilicet alchimistarum, artistarum, poetarum. In omnibus autem his scholis legitur transfiguratio. Prima schola est alchimistarum 597, 17rb Pro huius thematis introductione invenio tres scholas scilicet arborum et plantarum, vaporum et nebularum, dolentium animarum. Prima schola est plantarum et arborum in qua talis doctrina legitur quod quando arbores plorant 597, 37rb Pro huius thematis introductione invenio tres scholas scilicet Belzebub in Babylonia, Iani in Roma, Pharisaeorum in Ierusalem. Prima schola fuit in magna civitate Babyloniae in qua tractabatur 597, 9va Pro huius thematis introductione invenio tres scholas scilicet caeli, paradisi, mundi. Prima ergo schola fuit caeli. Nam in principio creavit Deus caelum et terram 597, 18vb Pro huius thematis introductione invenio tres scholas scilicet cordialem, pigmentalem, usualem. Prima est cordialis in qua legitur quod sanguis est summum elementum cordis 597, 35rb Pro huius thematis introductione invenio tres scholas scilicet dominorum, generatorum, magistrorum. Prima schola fuit dominorum et in ista et in aliis legitur quod omnes intendunt sibi assimilare 597, 27ra Pro huius thematis introductione invenio tres scholas scilicet elegantiae in naturalibus affluentiae in temporalibus eminentiae in dignitatibus 597, 20ra Pro huius thematis introductione invenio tres scholas scilicet Eremitarum, Praedicatorum, Minorum. Prima schola est Eremitarum in Aegidio qui videtur velle tenere quod si primus homo non peccasset 597, 7ra Pro huius thematis introductione invenio tres scholas scilicet extimam, intimam, extrinsecam. Prima schola dicitur extima et haec est excellentiae dignitatis et in hac lego trinitatem 597, 33rb Pro huius thematis introductione invenio tres scholas scilicet Hebraicam, Graecam et Latinam. Prima schola est Hebraica in qua considero Salomonem facientem iustitiam 597, 8rb Pro huius thematis introductione invenio tres scholas scilicet internas, alternas, subalternas. Prima est interna et est voluntatis et in hac legitur quod ea quae sunt in homine ipsi voluntati praesenti principaliori et digniori subalternas serviunt 597, 42ra Pro huius thematis introductione invenio tres scholas scilicet lapidum, herbarum, verborum. Prima ergo schola est lapidum et in illa legitur magna virtus. Nam videmus quod carbunculus 597, 30va Pro huius thematis introductione invenio tres scholas scilicet mechanicas, trepidas et incredulas. Primo invenio scholas mechanicas et iste sine dubio ut earum opera cognoscantur 597, 10vb 448 Pro huius thematis introductione Pro huius thematis introductione invenio tres scholas scilicet mechanicas, piraticas, maleficas. Prima schola est mechanica et haec utitur ligno 597, 43ra Pro huius thematis introductione invenio tres scholas scilicet motorum, auditorum, bellatorum. Prima est motorum et in illa legitur lux gratiae unde Gen. primo dixit Deus fiat lux et facta est lux 597, 36ra Pro huius thematis introductione invenio tres scholas scilicet organicam, scientificam, evangelicam. Prima schola dicitur organica et in hac legitur quod quis doctus esse non potest 597, 33va Pro huius thematis introductione invenio tres scholas scilicet planetae, cometae, monetae. Prima est planetae et in hac legitur vita secundum quod melior est constellatio vel planeta de tanto efficimur levioris vitae 597, 35vb Pro huius thematis introductione invenio tres scholas scilicet Sardanapalli in Asia, Catilinae in Roma et Attilae in Ungaria. Prima ergo schola est Sardanapalli 597, 22va Pro huius thematis introductione invenio tres scholas scilicet sapientiae, praesentiae, refulgentiae. Prima schola est sapientiae in qua legitur quod sapientium nomen clarificatur per famam in multis locis 597, 39va Pro huius thematis introductione invenio tres scholas scilicet Thebanam, Troianam, Romanam. Prima schola fuit Thebana et in hac pro sui tempore lecta fuit divisio 597, 28ra Pro huius thematis introductione invenio tres scholas scilicet urbis fundatorum, in campo bellatorum, in campo clericorum. Prima schola est urbis fundatorum 597, 38rb Pro huius thematis introductione invenio tres scholas scilicet veneficas, angelicas, truffaticas. Prima schola est angelica et in hac legitur magna opera facta fuisse et primo legitur de archangelo Michaele 597, 36vb Pro huius thematis introductione invenio tres scholas scilicet vulgaricam, canonicam, theologicam et in his legitur quod nullus possit iudicari 597, 32ra Pro huius thematis introductione invenio tres scholas scilicet vulgarium, naturalium, moralium. Prima mensuratur experientia secunda ratione tertia discretione 597, 2va Pro huius thematis introductione invenio tres scholas videlicet Origenis, Augustini, Hieronymi. Prima schola est Origenis cuius talis fuit opinio seu doctrina 597, 30rb Pro huius thematis introductione invenio tres scholas videlicet physicam Avicennae, cyrusicam Galeni, evangelicam Iesu Christi. In prima schola fuit magnus medicus naturalis et in illa legebatur 597, 13rb Pro huius thematis introductione invenio tres scholas. Prima igitur schola fuit Moysii inesse figurali 597, 1ra Pro huius thematis introductione invenio tres scholas. Prima schola est agricultorum qui abundat in gratia temporali cum tales fructus terrae et gratiam Dei 597, 16ra Pro huius thematis introductione invenio tres scilicet olivae in Minerva, lauri in Mercurio, palmae in Marte. Prima schola est olivae in Minerva. Nam ut possum capere ex dictis poetarum oliva consecrata est in nomine Minervae 597, 40vb Pro huius thematis introductione reperio tres scholas mirificam, munificam, magnificam. Prima schola est mirificum (!) et haec schola est Dei. Prope enim Deus qui fecit mirabilia 597, 32vb Pro huius thematis introductione reperio tres scholas scilicet elementalem, spiritualem, iudicialem. In his tribus scholis legitur mors. Prima ergo schola est elementalis quia composita originaliter ab his cum omne compositum resolvi 597, 34vb Pro huius thematis introductione tres invenio scholas scilicet Medeae, Dyodonis, Ysiphilae. Prima ergo schola fuit Medeae nam Medea fuit filia Ochis (!) regis qui regnavit in insula Colchide 597, 25va Pro huius thematis introductione tres scholas invenio scilicet aequitatis, benignitatis, sanctitatis. Prima schola est aequitatis et in hac legitur quod relevetur infirmus 597, 34va Pro huius thematis introductione tres scholas invenio scilicet examinationis, confessionis, obligationis. Prima schola est examinationis et in ista schola legitur quod reus dato quod omnia sunt allegata et approbata 597, 34ra Pro huius thematis introductione tres scholas invenio scilicet filiorum, inimicorum, discipulorum. Prima schola est filiorum. In hac considero inter filios primi parentis 597, 29ra Pro huius thematis introductione tres scholas invenio scilicet Martis, partis, mortis. Prima est Martis. Nam Mars ut pluries dictum est planeta malevolus est et nati sub illo sunt pessimi 597, 36va Pro huius thematis introductione tres scholas invenio scilicet nuptiarum, victoriarum et scientiarum. Prima ergo fuit nuptiarum et illam invenio in rege Asuero 597, 29rb Pro huius thematis introductione tres scholas invenio scilicet Polestinoris, Mesabellis, Pignaleonis. Prima schola fuit Polestinoris qui fuit rex cuiusdam insulae 597, 24rb Pro huius thematis introductione tres scholas invenio scilicet potentiarum, naturarum, grammaticorum. Nam prima est potentiarum et quia habemus plures potentias 597, 32rb Pro huius thematis introductione tres scholas reperio et fortunae, naturae, rationalis creaturae. Prima schola scilicet naturae dicit quod omnia naturalia 597, 6rb Pro huius thematis introductione tres scholas reperio ethicam seu monasticam, oeconomicam seu familiarem, politicam seu civilem. Prima schola est monastica in qua legitur 597, 14rb Pro huius thematis introductione tres scholas reperio scilicet Epicureorum, Platonicorum, Hypotheticorum. Sed in omnibus istis scholis doctrina legitur quae secundum aliquam partem merito nobis timorem 597, 5rb Pro huius thematis introductione – Quaesumus domine 449 Pro huius thematis introductione tres scholas reperio scilicet illustrium magnificorum, secunda eloquentium, tertia decipientium et proditorum. Prima schola fuit illustrium etc. 597, 12ra Pro huius thematis introductione tres scholas reperio scilicet naturae, scripturae, gratiae. Primo dico quod invenio scholam naturae 597, 4ra Pro huius thematis introductione tres scholas reperio videlicet mundi, carnis, daemonis. Prima schola est mundi in prima legitur apparentia sive existentia 597, 21rb Pro intellectu dubitatur quale fuit illud desertum ad litteram 53, 174va Pro intellectu litterae notandum ut narrat Iosephus 13 libro Antiquitatum capitulo 14 regnum Iudaeorum fuit interceptum 481 annis et duos menses a captivitate 53, 71ra Pro intellectu litterae notandum quod Iohannis 12 dicitur Quando Iudaei iam ex consilio Caiphae 53, 249va Pro intellectu notandum quod secundum magistrum historiarum Iohannes arguebat Herodem propter Herodiadem quam a fratre suo receperat 53, 26ra Pro intellectu notandum sicut habetur Matthaei 14 Postquam Christus transfretavit per mare Tiberiadis venit Capharnaum 53, 237rb Probet autem seipsum homo … (1 Cor 11,28). Aristoteles 2o De anima Simile a simili nutritur sicut patet in cibo digesto a quo corpus nutritur 153, 81rb Propitiare nobis quaesumus omnipotens pater per unigeniti tui corpus et sanguinem 353, 65r Propter multiplicationem humani generis Deus specialiter instituit matrimonium 53, 84vb Prosequendo sermonem iam inceptum tunc sciendum quod quanto caelum aperitur per intensam contritionem 153, 9rb Protector noster in te sperantium Deus sine quo nihil est validum 353, 54r Protege domine famulos tuos subsidiis pacis et beatae Mariae virginis patrociniis confidentes 405, 214r, 222r Protege domine populum tuum et apostolorum tuorum Petri et Pauli et omnium aliorum 353, 31r Protege domine populum tuum et apostolorum tuorum Petri et Pauli patrocinio confidentem 405, 218r Publii Ovidii Nasonis poetae vetusta equestris ordinis familia fuit 579, 1r Puer autem crescebat … (Lc 2,40). Ex quo in praesenti evangelio fit mentio de puero nobis recenter nato. Tunc pro praesenti sermone tria sunt dicenda. Primo quare Christus parvulus factus est 153, 15vb – Sicut puer Iesus crevit corporaliter ita et nos crescere debemus spiritualiter 153, 17va Puer Iesus proficiebat sapientia aetate (Lc 2,52). Praesens evangelium tangit statum Christi tempore pueritiae circa 12 annum 53, 81rb – Quando diligenter consideramus inaestimabilia divinae bonitatis beneficia nobis exhibita 53, 80va Pulcherrime Iesu Christe rogo te per illam sacratissimam effusionem pretiosissimi sanguinis tui quo sumus redempti 405, 224r Puppe saporatus iubet obmutescere natus 308, 228vb Quadam autem vice venerunt gentiles ut eum tenerent 308, 3rb Quadam vice antiquus hostis corporalibus eius 308, 54va Quadam vice cum in spiritu temptatus esset totum mundum laqueis sese invicem 308, 46ra Quadam vice cum praedicans Felix praedicaret et persecutor 308, 3ra Quadruplex agitur festum in hac septimana scilicet festum venerationis festum recreationis festum reconciliationis 617, 149r Quae procedunt de ore haec coinquinant hominem (Mt 15,18). Pro huius thematis introductione invenio tres scholas Catonis, Tullii et Senecae. Prima schola est Catonis et quamquam intra illam bona vita legatur tamen inter alia qui ibi leguntur 597, 29vb Quaecumque solveris super terram erunt soluta et in caelo (Mt 18,18). Pro huius thematis introductione tres scholas invenio scilicet nuptiarum, victoriarum et scientiarum. Prima ergo fuit nuptiarum et illam invenio in rege Asuero 597, 29rb Quaeritis me et non invenietis (Io 7,34). Pro huius thematis introductione invenio tres scholas scilicet motorum, auditorum, bellatorum. Prima est motorum et in illa legitur lux gratiae unum Gen. primo dixit Deus fiat lux et facta est lux 597, 36ra Quaeritur primo utrum et Nephiae et Typhones descendunt ad terram ex nubibus 440, 347ra Quaeritur quis effectus baptismi. Respondetur secundum Thomam in 4o Pueri quidam in baptismo mundantur 447, IXra Quaeritur utrum angelus in primo instanti creationis fuerit malus actu propriae voluntatis 52, 44ra Quaestio inter doctores scholasticos solet moveri utrum Deus posset non retribuere mercedem summae beatitudinis creaturae rationali diligenti eum fideliter 597, 43va Quaestio oritur inter doctores utrum in hac vita possit Deum videre facialiter 597, 44ra Quaesumus domine pro tua pietate miserere animae famuli tui a contagiis mortalitatis 654, 87r Quaesumus domine ut animae famuli tui cuius obitus sive diem commemoramus sanctorum tuorum largiri digneris consortium et rorem misericordiae tuae perennis infundere 14820, 85v 450 Quaesumus omnipotens Deus – Quidam habens uxorem Quaesumus omnipotens Deus ut beatae et gloriosae semperque virginis Dei genitricis Mariae intercessio gloriosa nos ab omni malo protegat 353, 161r Quam detestabile sit vitium invidiae et fugiendum patet propter multa mala 53, 240ra Quamquam nesciatur de homine quoquam an salvetur vel damnetur 153, 127ra Quamvis de astrolabii compositione tam modernorum quam veterum dicta habeantur 776, 61r Quamvis divina miseratio multis variisque modis in scripturis sanctis omnis vocare dignata est 617, 63r Quamvis haeretici miseri mentiantur carnale martyrium esse illicitum eorum tamen falsitas manifeste apparet 153, 32va Quamvis in nuptiis homines soleant convivia tenere haec autem caute tenenda sunt in Dei timore 53, 92rb Quamvis multis modis homo serviat Deo et pro peccatis suis satisfaciat 53, 338rb Quamvis venerandi domini haec pestis tam admirabilis et horrenda a causis superioribus procedat regulantibus et gubernantibus corpora nostra 748, 71r Quando aliquis se cupit ad communem vitam transferre 76, 19r Quando dignitatem festorum totius anni clarius inspicimus invenimus quod ipsa praesens paschalis sollemnitas 53, 310ra Quando diligenter consideramus inaestimabilia divinae bonitatis beneficia nobis exhibita 53, 80va Quando dorsum equi laeditur per sellam 160, 1r Quando enim occurrat festum alicuius sancti tunc consuevimus eius legendam et passionem ut Laurentii Petri etc. Sic enim hodie est flendum 174, 156ra Quando equus patitur fissuras contra ungulam vulgariter in den fisselen 160, 1r Quando equus patitur in pedibus defectum 160, 1r Quando feudum tenet a laico de iudice c. ceterum 145, 314va Quanta Iudaeis acciderint gravia post temeritatem crucis in Christum scribit magister in Ecclesiastica Historia libro 2o capitulo 6o dicens Primo quidem temporibus Tiberii refert in urbe Roma 166, 52va Quantum ad praesentem festivitatem possunt haec verba esse matris ecclesiae 617, 172r Quantum ad statum paenitentiae ostendit autem civitatem iustorum laudabilem duplici respectu 162, 9ra Quare autem non observamus nostrum ieiunium 52, 39vb Qui manducat hunc panem vivet in aeternum (Io 6,58) et in evangelio hodiernae lectionis. Magnam hodie et praecipuam carissimi omnique devotione recolendam agimus sollemnitatem ad cuius celebrationem venerandam venerabilis Hugo de Sancto Victore in quodam sermone ipsius sollemnitatis nos devote invitat 174, 166va – Loquendo de manducatione huius venerabilis sacramenti sciendum quod hoc sacramentum tripliciter sumitur 153, 158va Qui non est dives et nescit vivere pauper / Ille modum vitae nescit habere suae 165, 42ra Qui se exaltat humiliabitur (Mt 23,12). Pro huius thematis introductione invenio tres scholas scilicet elegantiae in naturalibus affluentiae in temporalibus eminentiae in dignitatibus 597, 20ra Quia Christus multa signa fecit in civitatibus Iudaeorum ipsi nihilominus in sua pertinacia permanserunt 53, 156vb Quia in tentivum (!) passionis Christi et occasio fuit proditio 53, 272va Quia nativitatem beatae Mariae agimus ideo gaudere debemus quia ipsa est stella 78, 91rb Quia nullus evangelistarum materiam dominicae passionis plene ac complete conscripsit 756, 1ra Quia plurimi ob nimiam quandoque accurationem et magnam scriptorum sententiam canones utilis astrolabii declarantes intellegere 776, 47r Quia postillatio verborum evangelii satis patet ex alio sermone facto de eodem. Ideo solummodo dicendum erit 53, 110vb Quia ut ait Ambrosius in sermone de Quadragesima Ecce nunc tempus acceptabile est in quo confessio a morte animam liberat 53, 197vb Quia video te diu vacare otio silentio 360, 1r Quicumque esset laesus vel percussus a malitia Mercurii 748, 54rb Quicumque vult salvus esse ante omnia opus est ut teneat catholicam fidem 190, 67rb Quid agendum sit dum quis habeat aliqua peccata a quibus ille cui confiteatur non potest absolvere dicit beatus Thomas 155, 15ra Quid etiam de illo monacho commemorem cuius nomen quia adhuc superest 166, 57ra Quid retribuam domino pro omnibus quae retribuit mihi panem caelestem accipiam de mensa domini 100, 28ra Quis ex vobis arguet me de peccato etc. ubi supra (Io 8,46). Christus dominus frequenter in suis sermonibus arguit Iudaeis super peccatis suis 53, 246va Quidam canonicus ad eum venit et ut ad monachum eum reciperet 308, 59rb Quidam egregius doctor in sermone publice praedicavit quod quicumque devota intentione infrascriptas missas legeret seu legi disponeret pro anima 754, 26v Quidam habens uxorem condemnatus de crimine capitali 614, 289v Quidam religiosus ordinis – Reverendo in Christo 451 Quidam religiosus ordinis Cartusiensis monasterii in Ruremunda in solemnitate purificationis gloriosissimae virginis Mariae in summa missa toto conventu praesente et inspiciente ab exterioribus sensibus 754, 11v Quo abiit dilectus tuus o pulcherrima mulierum (Ct 6,1). In his verbis tria videnda sunt scilicet quae sit dilecta quis sit dilectus quo et quomodo abiit. Circa primum sciendum quod dilecta est beata virgo 617, 170r Quo studio et quo affectu a nobis orandus sit Deus 755, 4ra Quod multum terribile sit extremum iudicium videtur posse trahi ex his quae in proximo sermone dixi 53, 11rb Quod quilibet homo et maxime Christianus debeat fugere lites et contentiones 53, 246ra Quodam tempore cum Remigius factus est Remensis archiepiscopus et in domo cuiusdam 308, 33vb Quomodo hic litteras scit cum non didicerit (Io 7,15). Pro huius thematis introductione invenio tres scholas scilicet organicam, scientificam, evangelicam. Prima schola dicitur organica et in hac legitur quod quis doctus esse non potest 597, 33va Quoniam apostolica sedes intendit providere negotiis non personis 145, 314va Quoniam autem natura. Hic incipit tertius liber Physicorum in quo Aristoteles determinat de passionibus 168, 17r Quoniam frequenter in foro paenitentiali dubitationes circa matrimonium 253, 88rc Quoniam gloriosus et excelsus Deus de largitate sua medicinam produxit et medicum velut naturae refugium creavit 748, 66v Quoniam quidem. Iste liber intitulatur liber Physicorum Aristotelis filii Nicometi Graecorum sapientissimi 168, 1r Quoniam quidem scire et intellegere contingit circa omnes scientias quarum principia sunt aut causae aut elementa 168, 1r Quoniam te carissime filii Iohannes lacrimas maesto cordis dolore perpetim effundere perspexi 748, 162ra Quoniam ut ait Galenus septimo therapeuticae methodi auctoritate Platonis opus in aliqua scientia vel arte 748, 74r Quoniam ut ait Hieronymus sacerdotis officium est respondere de lege et ut dominus dicit per Malachiam prophetam Labia sacerdotis custodiunt sacramentum et legem 447, 1ra Quoniam velut quattuor paradisi flumina … Scriptura sacra de duobus agit … Veteris ac novae legis continentiam considerantibus 532, 34v Recipe lixinii partes tres olei nucum partem unam et valet contra cancrum et lupum et ignem persicum in principio certissimum est 748, 54ra Recipe tormentillae drachmas quindecim terrae sigillatae 748, 72r Redde illis mercedem (Mt 20,8). Quaestio inter doctores scholasticos solet moveri utrum Deus posset non retribuere mercedem summae beatitudinis creaturae rationali diligenti eum fideliter 597, 43va Refert Gregorius in registro. Legitur quod sanctus Augustinus nec cum sorore 308, 14rb Regem apostolorum dominum venite adoremus 154, 4r Regem manum. Adoremus dominum 405, 1r Regulus obnatum stat supplex febre gravatum 308, 228vb Religiosis atque honestis viris in Christo dilectis fratribus studentibus Neapolim 125, 1ra Reminiscere miserationum tuarum quaesumus domine et famulos tuos 353, 62r Repetiturus decretalem [X.3.41.1.] presbyter quae prima sub rubrica de celebratione missarum situatur et ponitur 155, 1ra Replebimur in bonis domus tuae sanctum est templum tuum mirabile in aequitate Graduale Neocellense I, 324r Repleti sunt omnes spiritu sancto (Act 2,4). Aristoteles 9o Metaphysicae Contraria non fiunt in eodem. Cum ergo spiritus sanctus intrat in cor hominis tunc mala et peccata depellit 153, 137ra – Ex quo iam illis sacris diebus peragitur missio spiritus sancti tunc aliquid de donis spiritus sancti pro praesenti sermone dicendum est 153, 141rb Respice quaesumus omnipotens et misericors Deus super animas omnium fidelium defunctorum 654, 40r Respondens Iesus dixit siehe In illo tempore respondens Iesus dixit Respondit Thomas dixit Deus meus et dominus meus (Io 20,28). Verba praedicta quilibet potest dicere qui digne in hoc sacro tempore communicavit 153, 106ra Responsorium de ascensione nostri domini 483, 53v Restat quaerere aliqua circa tertium librum et verum est quod ab antiquo solemus illum totalem librum dividere in quattuor partiales 440, 347ra Reverendissimo in Christo patri ac domino domino Sebastiano Dei gratia episcopo Brixinensi necnon egregiis eximia doctrina et sapientia eiusdem ecclesiae praelatis ac canonicis Paulus Ursinus philosophus et medicus salutem dicit. Quae vitio nostro non proveniunt quaeque in nostra minime sita sunt 694, 1v Reverendissimo in Christo patri fratri Hugoni magistro ordinis fratrum Praedicatorum dignissimo ac sacrosanctae paginae professori. Eius subditus frater Alfonsus Bonihominis Hispanus promissam oboedientiam 617, 117r Reverendo in Christo patri ac domino domino Raimundo divina providentia sanctae sedis Valenciae episcopo servorum devotorum minimus Guido de Monte Rocherii cum devota 550, 1ra 452 Reverendo in Christo – Sciendum quod secundum dictum Reverendo in Christo patri et domino suo domino N. Dei gratia sancti Adriani diaconi cardinalis Laurentius dictus Kycius (!) mereschaltus (!) de Urbe … Inter cetera animalia quae ad dignitatis humanae solacium 160, 4ra Rex Assuerus tertio anno imperii sui fecit grande convivium cunctis principibus et pueris suis et vocavit omnem populum qui inventus est in Susia (Est 1,3). Per hoc convivium Veteris Testamenti figuratum est nobis convivium a Christo hodie institutum sacri corporis et sanguinis sui 617, 173v Rex David cooperto capite etc. 2 R. (CAO Nr. 4650). Historia haec est de David quomodo planxit filium suum Absolon. Historia nota est scilicet quid significat 617, 163v Rex pacificus praemissa salutatione sic pone casum in ista constitutione. Rex pacificus id est dominus noster Iesus Christus 155, 106ara Rheuma est fluxus humorum ad partes subiectas. Rheumatis autem tres species ab auctoribus distinguuntur 123, 215v Roma vetus veteres cur (!) te rexere Quirites 579, 14v Rustica deflenti puero iuraverat olim / Ni taceat rapido quod foret esca lupo 155, 155v Sacerdos qui sine mortalibus est peccatis et bono proposito si non celebrat missam cum possit et habet copiam celebrandi 167, 71va Sacramentum mei corporis sumite quo ad fideles et conficite quo ad presbyteros 53, 258rb Sacratissimam domine dilectissimi filii tui passionis memoriam devote recolentes pro liberatione animarum 654, 43v Salvator Christus in praesenti evangelio movet singulos homines ad considerandum sua propria opera 53, 130ra Salvator clementissime qui inter opprobria quae tulisti piae matris anxietatibus compassus fuisti 353, 113v Salvator noster Christus dominus saepius verbis suis nos admonet non solum mala fugere 53, 142rb Salvator noster Christus dominus sicut omnia in sua passione ostendit ut declararet suam humanitatem 53, 318rb Sancta Dei genitrix perpetua virgo Maria cum omnibus sanctis et electis diei intercedere dignare 405, 127v Sancta trinitas unus et omnipotens Deus nos quaesumus a peccatorum nostrorum maculis 353, 122v Sancte Deus sancte fortis sancte et immortalis Deus miserere animabus omnium fidelium defunctorum 405, 220v Sancti patres primitivi Christianae ecclesiae instituerunt ieiunium fidelibus observandum 53, 176vb Sancti spiritus corda nostra mundet infusio 353, 39r Sancti spiritus gratia suffragante de personis 7, 32va Sanctus Ambrosius eo die missam celebrans super altare 308, 29ra Sanctus Gregorius in prologo orationis dominicae dicente Praeceptis salutaribus moniti et divina institutione formati audemus dicere 161, 8va Sanctus Thomas Cantuariensis bonae indolis adolescens cum adhuc in scholis 308, 187vb Sanctus Udalricus praesul Augustensis cum quandam ecclesiam suae dioecesis dedicare vellet 308, 188va Sanguinaria id est bursa pastoris sive succus 748, Ir Sanus fiebat a quacumque detinebatur infirmitate (Io 5,4). Pro huius thematis introductione invenio tres scholas videlicet physicam Avicennae, cyrusicam Galeni, evangelicam Iesu Christi. In prima schola fuit magnus medicus naturalis et in illa legebatur 597, 13rb Sapiens Ecclesiastes 3 (1 und 4) dicit quod omnia tempora tempus habent. Est tempus flendi et tempus ridendi 53, 280rb Sapientia aeterna splendor gloriae et substantia patris qui universa de nihilo creasti 353, 14r Scandit montana pia virgo stupet veterana 308, 228rb Sciendum est quod moderno tempore in Apulia in provincia Capitinate 125, 198va; 165, 71rb Sciendum primo quod Christus servavit cicatrices vulnerum in suo corpore glorificato 153, 104va Sciendum quod ab origine mundi non fuit tale festum in caelo nec umquam usque post iudicium erit 153, 126ra Sciendum quod Christus die octavo circumcisus est et sanguinem suum fudit 153, 21ra Sciendum quod Christus voluit temptari ut ostenderetur quod nos spiritualiter pugnare oporteret 153, 65rb Sciendum quod circa praedicta verba tria sunt dicenda. Primo quomodo Deus trahit et vocat hominem 153, 144rb Sciendum quod Decretum dividitur in tres partes principales. Quarum prima vocatur Distinctiones 614, 273v Sciendum quod Deus creavit primos parentes iustos iustitia naturali id est sine omni obliquitate 53, 63va Sciendum quod dicit Haimo Mons in quo Deus sedet caelum intellegitur 153, 37va Sciendum quod hodie festum trinitatis institutum est ut omnes fideles agnoscant unam simplicem deitatem et trinitatem 153, 147va Sciendum quod hodierna festivitas inter alia festa dignitatem singularem obtinet 153, 134va Sciendum quod in caelo sunt solummodo boni in inferno solummodo mali 153, 44va Sciendum quod per pastorem in praesenti evangelio intelleguntur praelati et per oves intelleguntur subditi 153, 107vb Sciendum quod plura genera hominum sunt qui dicunt verbis se credere in Deum 153, 115va Sciendum quod praesens festivitas praecellit alias festivitates in sollemnitate et iucunditate 153, 89va Sciendum quod secundum dictum Salomonis stultorum infinitus est numerus 153, 99rb Sciendum quod – Sicut dicit Gregorius 453 Sciendum quod sicut Deus arguet peccatores de suis excessibus in futuro 153, 117vb Sciendum quod sicut Iudaei olim tenuerunt se ad discipulos Christi scilicet separando eos a participatione et communione ipsorum 153, 131vb Sciendum quod triplex est nativitas naturalis spiritualis et caelestis 153, 150va Sciens Iesus quia venit hora eius ut transeat ex hoc mundo ad patrem (Io 13,1). Dominus Iesus Christus et magister noster pretiosum nostrae redemptionis pretium 53, 272rb – Quia in tentivum (!) passionis Christi et occasio fuit proditio 53, 272va Scire debetis in Christo dilectissimi quod dies hodierna ex salvatoris nostri nativitate nobis redditur gaudiosa 153, 4va Scire debetis in Christo dilectissimi quod sancta mater ecclesia catholica hodierna die duo nobis tempora repraesentat 153, 48rb Scito quod talia oportet te praeparare qualia sunt in hostia Quanto maior est festivitas tanto se melius homines praeparare debent 177, 75rb Secundo idem angelus octo officia exercuit circa filium virginis declarandum. Primum erat officium docendi unum Ioseph dubitantem 31, 54ra Secundum Bedam Satorem istum nullum melius quam Dei filium intelligere possumus 53, 142vb Secundum Chrysostomum homilia 24 super Matthaeum Ne forte diceret populus apud se 53, 97ra Secundum dictum Augustini in domo Dei id est in ecclesia quattuor sunt necessaria scilicet grammatica musica computus et ius canonicum. Grammatica ad scripturas sacras intelligendum musica ad laudes Dei decantandum 167, 1ra Secundum doctores adventus domini agitur per quattuor hebdomadas ad denotandum quod est quadruplex adventus filii Dei 53, 1va Secundum historiam post mortem Iulii Caesaris qui a poetis commendatur 53, 47vb Secundum ordinem naturalem in mundo corpora magis conformia in natura sint magis propinqua in situ 31, 79vb Secundum Originem Iudaei videntes prope esse tempus adventus Messiae 53, 30va Secundum quod dicit Beda in homilia de Christi transfiguratione ut patet Matthaei 17 Salvator humani generis Christus dominus 53, 238va Secundum scripturas sanctorum homo per peccatum fit instabilis et mutabilis et despectus 53, 151va Sed etiam volo quinque verba sensu meo loqui ut et alios instruam (1 Cor 14,19) 155, 15ra Semen est verbum Dei (Lc 8,11). Est enim semen verbum Dei quod Christus de caelo attulit in terra seminavit 153, 54ra Semper nos domine martyrum tuorum Nerei et Achillei foveat 14820, 85v Sequere me (Io 21,19). Verba proposita sunt a Christo beato Petro dicta et iam dicuntur cuilibet homini qui vult salvari 153, 11va Sequere me (Mt 9,9). In illo evangelio duo notantur scilicet dignitas vocantis cum dicitur Sequere me 617, 187Iv Si alicui plus placuerit praedicare de evangeliis quam de epistolis 174, 1ra Si per alidadam horariam vis horam diei invenire naturalem poni alidadam super altitudinem mediae diei illius in dorso astrolabii suspensi 776, 71v Si quis princeps vel rex vel magnus dominus vult destinare nuntium vel legatum ad summum principem vel regem vel dominum magnum pro magna et ardua causa 31, 67rb Si quis revelet peccatum suum sacerdoti non ut paeniteat sed ut consilium et auxilium ab eo recipiat 155, 9va Si uxor alicuius viri quia non potest prolem habere timens quod possessio viri ad alios devolveretur 155, 7vb Si vixeris secundum carnem / morieris morte aeterna 397, VDS Si volumus amplius scire quod multipliciter est sanctissima virgo domina Maria benedicta inspiciamus 31, 89rb Sic ambuletis ut abundetis magis prima ad Thessal. 4 (1). Pro huius thematis introductione invenio tres scholas. Prima schola est agricultorum qui abundat in gratia temporali cum tales fructus terrae et gratiam Dei 597, 16ra Sic comedetis illum renes vestros accingetis calciamenta vestra habebitis in pedibus (Ex 12,11). Haec verba licet ad litteram sint dicta de agno paschali cuius immolatione 53, 251va – Nunc igitur iuxta praemissa sicut dixi ad contemplationem 53, 252ra Sic legimus quod unigenitus Dei filius volens [ 153, 156vb Sic mundo pactum solvit verbo caro factum 308, 228rb Sicut apotheca est salubris ita musica delectabilis quia dulcedine sua 776, 1v Sicut auris non potest satiari auditu nec oculus 53, 265va Sicut credidisti fiat tibi (Mt 8,13). Pro huius thematis introductione invenio tres scholas scilicet vulgarium, naturalium, moralium. Prima mensuratur experientia secunda ratione tertia discretione 597, 2va Sicut Deus disposuit omnia sapienter sic fidei incrementum et suam humanitatem 53, 70va Sicut dicit Aristoteles 4o Ethicorum Terribilissimum omnium terribilium videtur esse mors 153, 154ra Sicut dicit Bernardus in sermone de epiphania Priusquam appareret Dei humanitas latebat 53, 47va Sicut dicit Gregorius homilia 2 super evangelia Miracula domini et salvatoris nostri sic accipienda sunt 53, 174ra 454 Sicut enim – Suppliciter te Deus Sicut enim tunc vos scitis, quod mos praedicatorum est in principio sermonum praedicatorum ad beatam virginem recurrere 174, 156Ira Sicut in bello corporali securius pugnatur et citius obtinetur triumphus contra hostes 53, 169rb Sicut in dominica proxima futura recolitur spiritus sancti missio 153, 129va Sicut inter virtutes morales quae pertinentes sunt ad mores exteriores praecipue praecipua est gratitudo 53, 156rb Sicut omnipotens Deus verax est in praemissis Ita et misericors 53, 43rb Sicut ovis ad occisionem ductus est (Is 53,7). Hodie carissimi dies celebratur insignis et festiva in tanto maioris devotionis affectu a singulis Christi fidelibus est celebranda 174, 163ra Sicut patrifamilias placet servus qui parate exsequitur suam voluntatem 53, 129va Sicut puer Iesus crevit corporaliter ita et nos crescere debemus spiritualiter 153, 17va Sicut tactum est in carta quadam hoc in loco reperta 76, 71Ir Sicut valde consultum est servo procuratori alicuius potentis domini qui numquam rationem fecit 53, 19rb Signa autem eos qui crediderint. Haec sequentur (Mc 16,17). Quamquam nesciatur de homine quoquam an salvetur vel damnetur 153, 127ra Signa generalia putridarum eis propria sunt duo ut dicit Averroes 748, Iv Signum non dabitur ei (Mt 12,39). Pro huius thematis introductione invenio tres scholas scilicet mechanicas, trepidas et incredulas. Primo invenio scholas mechanicas et iste sine dubio ut earum opera cognoscantur 597, 10vb Simile a simili nutritur sicut patet in cibo digesto a quo corpus nutritur 153, 81rb Simile est regnum caelorum homini patrifamilias (Mt 20,1). Chrysostomus super Matthaeum homilia 28 Homo Christus paterfamilias cui caeli et pater (recte: terra) una est domus familia 53, 124va – Dominus Deus creavit hominem immo et totum mundum propter suam bonitatem 53, 124ra – Ex quo in praesenti evangelio fit mentio de patrefamilias qui conduxit operarios 153, 50ra – Salvator Christus in praesenti evangelio movet singulos homines ad considerandum sua propria opera 53, 130ra – Scire debetis in Christo dilectissimi quod sancta mater ecclesia catholica hodierna die duo nobis tempora repraesentat 153, 48rb – Sicut patrifamilias placet servus qui parate exsequitur suam voluntatem 53, 129va Similiter autem. Hic est quartus liber Physicorum in quo Aristoteles determinat de passionibus 168, 24v Simpliciores et minus expertos confessores de modo audiendi 532, 125r Sinite utraque crescere … (Mt 13,30). Sciendum quod in caelo sunt solummodo boni in inferno solummodo mali 153, 44va Sistitur hic domino redimendus turture bino 308, 228va Sit per te excusabile quod per te gerimus 405, 210r Solent homines in novo anno munuscula sibi invicem dare 153, 19rb Solus Deus dicitur immortalis quia solus est incommutabilis nam et omnia (recte: anima) moritur dum de ore autem (recte: deserente) Deo de bono in malum mutatur 52, 4ra Solvite templum hoc et in tribus diebus excitabo illud (Io 2,19). Pro huius thematis introductione invenio tres scholas scilicet extimam, intimam, extrinsecam. Prima schola dicitur extima et haec est excellentiae dignitatis et in hac lego trinitatem 597, 33rb Spiritus eius ornavit caelos et obstetricante manu eius eductus est coluber tortuosus (Iob 26,13). Creaturarum consideratio pertinet ad theologos et philosophos 86, 1ra Spiritus est vapor temperatus genitus ex parte subtiliori sanguinis 748, 218vc Spiritus meus super mel dulcis (Sir 24,27). Ex quo spiritus sanctus est fons totius dulcedinis tunc quaeritur quare Deus quandoque dilectis suis subtrahit in praesenti consuetam gratiam 153, 139ra Stabat mater dolorosa iuxta crucem lacrimosa dum pendebat filius 353, 162r Stabunt iusti in magna constantia etc. (Sap 5,1). Circa quem textum est dicendum an sancti in aeterna beatitudine quam habebunt post resurrectionem sedebunt vel stabunt 53, 346vb Stephanus vidit caelos apertos (Act 7,56). Aristoteles 1o Ethicorum finis per quem omnia desiderantur et sperantur optimus est 153, 7rb Stetit angelus iuxta aram templi habens thuribulum aureum in manu sua 353, 3r Subveniat nobis domine misericordia tua ut ab imminentibus peccatorum nostrorum periculis 353, 47r Subsequens oratio de beata virgine et Dei genitrice Maria continens principaliora ipsius et filii eius gesta 654, 117r Summum constitue rescriptum post aliud consue 145, 1r Sunt autem septem genera hominum qui Deum non colunt sed idola adorant. Primus genus est de quibus 308, 210ra Supplicationem servorum tuorum Deus miserator exaudi ut qui in commemoratione sanctae Dei genitricis et virginis Mariae congregamur eius intercessionibus 353, 157v Suppliciter te Deus pater omnipotens qui es creator omnium rerum 654, 65v Surrexit non est – Unde Iohannes de Sancto Geminiano 455 Surrexit non est hic (Mc 16,6). Quando dignitatem festorum totius anni clarius inspicimus invenimus quod ipsa praesens paschalis sollemnitas 53, 310ra Tangere qui gaudes meretricem qualiter audes / palmis pollutis regem tractare salutis 167, 72r Te Deum laudamus te dominum misericordissimum confitemur tuam maximam misericordian semper imploramus 353, 42v Te domine sancte pater omnipotens aeterne Deus supplices deprecamur pro animabus omnium fidelium defunctorum quas ab origine mundi accersiri voluisti 654, 36v Te invocamus te adoramus te laudamus beata trinitas miserere nobis 353, 117r Te lucem veram et lucis auctorem domine deprecamur ut digneris a nobis tenebras depellere vitiorum 405, 92r Te matrem laudamus te Mariam virginem confitemur 353, 168v Templum Dei sanctum est quod estis vos si quis autem templum Dei violavit disperdet illum dominus Deus (1 Cor 3,17). In verbis istis apostolus duo facit, ostendit enim fidelis animae sanctitatem et exsecratur sanctitatis violatorem 162, 8ra Templum hoc sanctum ingredientes rex pie exaudi nos domine. Nota tria templa placentia Deo. Primum templum est aureum secundum est eburneum tertium est lapideum 162, 3ra Tempore pestilentiali si vis ut apostema ex materia venenosa 748, 70v Thales dicit Antiquissimum entium est Deus ingenitum et in aevum. Optimum entium mundus nam opus Dei 155, 144ra Thomas autem non fuit cum eis quando venit Iesus (Io 20,24). Notandum quod beatus Thomas Christum discipulis apparentibus non vidit quia non fuit cum eis sed post octo dies eum vidit 153, 103va Tibi dico surge (Lc 7,14). Pro huius thematis introductione tres scholas invenio scilicet aequitatis, benignitatis, sanctitatis. Prima schola est aequitatis et in hac legitur quod relevetur infirmus 597, 34va Tota summa huius evangelii est quod Christus voluntarie ivit ad desertum locum 53, 170ra Totam vitam nostram plenam fecit Deus temptationibus et etiam rationabiliter Deus permittit electos suos temptari 53, 164ra Tradetur enim gentibus … (Lc 18,32). Ubi sciendum quod passio Christi quae in hodierno evangelio nobis proponitur retrahere nos debet ab illicitis gaudiis huius saeculi 153, 58ra Transfiguratus est ante eos (Mt 17,2). Pro huius thematis introductione invenio tres scholas scilicet alchimistarum, artistarum, poetarum. In omnibus autem his scholis legitur transfiguratio. Prima schola est alchimistarum 597, 17rb Translati sumus de morte ad vitam (1 Io 3,14). Nos enim videmus ad sensum quod quando aliqui evaserunt mortem qui fuerunt in bello fortissimo ubi multi mortui fuerunt ad suos dilectos veniunt et dicunt 174, 90vb Transmutatur autem. In ista parte Aristoteles ponit quintum librum Physicorum in quo Aristoteles determinat de divisione motus 168, 39r Tres sunt omnes doctrinae quae ordine habentur etc. Istius libri qui Tegni id est Pars (!) parva dicitur 453, 33v Tres sunt omnes doctrinae quae ordine habentur. Prima quidem ex finis notione 453, 1r Triplex causa febrem generat custodit et auget 748, 1r Triplex est rigor scilicet praecedens et est in interpolatis coincidens ex multitudine materiae 748, 40v Tu domine qui nosti corda hominum etc. (Act 1,24). Hodie in ecclesia agitur de duabus personis scilicet de Iuda qui est de honore suo detrusus 78, 35ra Tu puer propheta altissimi vocaberis praeibis enim ante faciem domini parare vias eius (Lc 1,76). Nota quod est triplex via. Prima in mundum, secunda per mundum, tertia extra mundum. Prima via in mundo (!) est lamentabilis 78, 64vb Tua nos domine Deus misericordia ab omni surreptione vetustatis 353, 50v Tui amoris gratia mi Antoni Magliane ad hunc deveni locum 748, 1v Tulerunt ergo lapides … (Io 8,59). Aristoteles 8o Ethicorum dicit Vera amicitia habet reamationem. Et quia Christus vere nos dilexit 153, 77ra Tussis est motus naturalis ex pulmone 123, 218r Tutela bona confessionis quando confitetur mulier se habere filium spurium inter legitimos. Officii mulier quae partum supponit alienum vel de adulterio concepit 155, 7ra Ubi sciendum est quod per mare intellegitur praesens mundus et per motum magnum 153, 41va Ubi sciendum quod passio Christi quae in hodierno evangelio nobis proponitur retrahere nos debet ab illicitis gaudiis huius saeculi 153, 58ra Ubi sciendum quod seminator est Deus omnipotens qui exiit seminare semen suum id est genus humanum creando 153, 51vb Unde Iohannes de Sancto Geminiano dicit Conveniens est ut sentiat membrum quod sentit caput 153, 84ra 456 Unde Iohannes de Sancto Geminiano – Verba sunt beati Tobiae Unde Iohannes de Sancto Geminiano Videmus enim quod cum animal brutum sentit se vulneratum sollicite quaerit et procurat sanationis remedium 153, 60va Ure igni sancti spiritus renes nostros et corda nostra domine 654, 65v Urina est colamentum sanguinis et aliorum similiter humorum de naturae quidem actionibus natum 748, 138ra Usurarius quidam cum crux in quibusdam partibus praedicaretur cum ceteris caecus non ex devotione sed ex importuna circumstantium ammonitione 308, 194va Usurarius quidam dives nimis diversarum 308, 195rb Ut in aliis vero dominicis. Propter baptismalia sacramenta in tribus personis 483, 54r Ut quid Mercuri hic ades galero redimitus et citharam gestans 694, 3r Ut seriem mente teneas textus doceam te / Est Genesis primus, post Exodus ipse secundus 166, IIv Ut tuam domine misericordiam consequamur fac nos per intercessionem 654, 15r Utrum autem factus. Iste est octavus liber Physicorum in quo Aristoteles determinat de perpetuitate motus 163, 15r Utrum corpus discrasiatum ex generatione et tempore debeat reduci ad temperamentum 453, 54va Utrum de passionibus et operationibus animatorum ponenda sit scientia distincta a scientia de anima de animalibus et a scientia de plantis 440, 1ra Utrum habens confessionale a sede apostolica secundum formam consuetam possit sibi rationem istius eligere in confessorem quemcumque sacerdotem non suspensum 155, 14va Utrum in omni tempore morbi possit fieri evacuatio in febre putrida. Quod sic auctoritate illius aphorismi. Incohantibus morbis etc. 453, 53va Utrum ratione confessionalis quod dari consuevit possit quis absolvi in casibus episcopalibus a quolibet sacerdote 155, 14vb Utrum religiosus professus et divina praecepta cum substantialibus regulae conservans 76, 11r Utrum sacerdos in confessione eam absolvere posset cum qua peccavit fornicariae 155, 14vb Utrum sit possibile umorem vel umores nostri corporis peccare solum in quali. Et arguitur quod non. Primo sic quilibet umor peccans 453, 143ra Utrum solus Deus sit adoratione latriae adorandus 165, 131v Utrum tantum tres sint omnes doctrinae ordinariae. Arguitur quod non. Primo quot sunt quaestiones tot sunt scientiae vel doctrinae 453, 187ra Utrum usura mortaliter culpabilis sit penes intentionem principaliter mensuralis (!). Arguitur quod non quia usura attenditur penes pactum mutui inter diversas personas contractum ergo non penes intentionem principaliter 447, 207ra Vado ad eum qui me misit (Io 16,5). Divina dispositione et providentia ab aeterno Deus piissimus disposuit postquam Iudaicus populus 53, 333rb Vado piscari (Io 21,3). Ex quo hodie est ultima dies celebris praesentis festivitatis paschae et modo homines redire debent ad pristinos labores suos 153, 101ra Vehementiores fiunt motus aegrium et acta eorum in nocte 748, Iv Venerabilibus in Christo patribus dominis Dei gratia archiepiscopis et episcopis sedis apostolicae gratiam et communionem habentibus 253, 83v Venerabilis domine N. Vos misistis ad me pro Amigdalis ego deberem nos providere cum Amigdalis ego locutus sum cum domino N. 617, 195r Venerandae ac praedilectae pater ad satisfaciendum oboedientiae mihi per paternitatem vestram nuper iniunctae sub actu visitatione videlicet ut scribere vobis deberem quid sentiam 754, 13r Venit enim filius hominis quaerere et salvum facere (Lc 19,10). Hic sumat peccatrix anima confidentiam tripliciter. Quia Christus venit quaesivit et salvum fecit venit nascendo quaesivit praedicando salvum fecit moriendo 162, 7rb – Ista scribit beatus Lucas et sunt verba nostri salvatoris suam dilectionem quam ad quendam humanum habuit 162, 1ra Venite mittamus lignum in panem eius (Ier 11,19). Pro huius thematis introductione invenio tres scholas scilicet mechanicas, piraticas, maleficas. Prima schola est mechanica et haec utitur ligno 597, 43ra Vera amicitia habet reamationem. Et quia Christus vere nos dilexit 153, 77ra Vera patientia quanta bona et quantas utilitates afferat homini possumus ex diversis scripturis sacris colligere 53, 138va Verba praedicta quilibet potest dicere qui digne in hoc sacro tempore communicavit 153, 106ra Verba proposita leguntur in evangelio hodierno et per ea insinuatur nobis gloria praemium omnium sanctorum quorum hodie festivitas celebratur 617, 189r Verba proposita possunt esse interioris hominis id est animae respondentis ad quaestionem sibi factam 617, 171r Verba proposita sunt a Christo beato Petro dicta et iam dicuntur cuilibet homini qui vult salvari 153, 11va Verba sunt beati Tobiae ad vocem haedi cuiusdam balantis caprarum 53, 209rb Veri praedicatores – Zamay non spiero de veder 457 Veri praedicatores praecipue quibus non sunt certae elemosinae assignatae libentius vadunt ad eos convertendos qui nondum ad fidem Christi conversi sunt quam ad illos qui iam conversi sunt 155, 16vb Verus ac pius samaritanus dulcis Iesus qui vinum et oleum fundit 550, 44vb Videntes stellam gavisi sunt gaudio magno valde etc. (Mt 2,10). Glossa non fuit satis evangelistae dicere gavisi sunt sed addit valde magno gaudio 153, 25vb Videte ne furtivus sit reddite eum […] dominis suis quia non licet vobis edere ex furto aliquid aut contingere (Tb 2,13). Verba sunt beati Tobiae ad vocem haedi cuiusdam balantis caprarum 53, 209rb Vidi alterum angelum descendentem de caelo amictum nube et iris in capite eius et facies eius sicut sol (Apc 10,1). In verbis istis tria notantur de sancto Bernhardo. Primo sanctitas vitae cum dicitur Vidi alterum angelum descendentem de caelo amictum nube. Primus enim angelus fuit beatus Benedictus 78, 80ra Vidi civitatem sanctam Ierusalem (Apc 21,2). Dicit beatus Augustinus quod duplex est civitas 162, 8vb – Quantum ad statum paenitentiae ostendit autem civitatem iustorum laudabilem duplici respectu 162, 9ra Vidit Iacob in somnis scalam et angelos Dei descendentes et ascendentes per eam (Gn 28,12). Ait vere dominus est in loco isto et ego nesciebam (Gn 28,16). His verbis possumus quattuor notare. Primo quis sit Iacob secundo quod dormivit tertio quae sit scala quarto quod angelos vidit 162, 3va Vidit Iacobus scalam a terra usque ad caelum attingentem per quam ascendebant 181, 1r Virorum illustrium praesulum scilicet ac doctorum praeconia (!) sacrosanctam quintae huius distinctionis series continet et declarat nugas nimirum respuens 308, 12rb Vita brevis ars vero longa tempus acutum experimentum vero fallax iudicium autem difficile 453, 39r Vita brevis ars vero longa tempus acutum. Praetermissis quibusdam superfluis inquisitionibus in principio huius libri 453, 59ra Vobis datum est nosse mysterium regni Dei (Mt 13,11). Quaestio oritur inter doctores utrum in hac vita possit Deum videre facialiter 597, 44ra Vor allen dingen ir lieben prüder habt got lieb dar nach den nächsten. Wan die zway gepot synd uns vorderleich gegeben 146, 1r Vos autem contristabimini (Io 16,20). Hic est dicendum quare Deus permittit aliquos bonos homines ita acerbissime tribulari in praesenti 153, 115rb Vos estis autem genus electum regale sacerdotium gens sancta etc. (1 Pt 2,9). Haec verba sunt beati Petri ad commendationem et dignitatem sacerdotalem. In verbis praemissis possumus notare triplicem gradum sive excellentiam cuiuslibet sacerdotis. Primo eius dignitatem 597, 58ra Vos portae pulcherrimae quae in magnam surrexistis altitudinem 654, 3v Wer begert zu ainem donatum aufgenommen werden, der sol vor wissen, daz er nit zu müssichait und rue 76, 73r Zachaee festinans descende (Lc 19,5). Nota quod quidam Deum recipiunt in domum suam et non in animam quidam in animam et non in domum quidam in animam et in domum quidam nec in animam nec in domum 162, 1va Zachaeus interpretatur iustus sed quia ille iustus non erat ideo dominum videre non poterat. Sunt autem tres causae quare dominum videre non poterat. Primo propter divitias 162, 4ra Zamay non spiero de veder vendeta / de quella falsa lengua maledeta 4, Iv 458 UNVOLLSTÄNDIGE TEXTANFÄNGE (INCIPIT MUTILE) ] adhuc super corda Iudaeorum positum et ideo non valent videre splendidum legis sensum 483, 39r ] aenigma vel obscuritas sive imperfectio 86, 34ra (aeter)]nitas. Per te sumus redempti summa tu caritas Graduale Neocellense I, 221r ] aqua de floribus fabarum aut aqua furfuris aut aqua 748, Ir ] Aristoteles 2o Ethicorum cum dicit eam esse non contemplationis gratia. Iste est prologus quem praemittit magister Buridanus libris Ethicorum quo primo innuit primam conclusionem 440, 89ra ] Ausoniis disterminat arva colonis unde apparet Antenorem non ad Italiam venisse 173, 27ra ] caelis et quidcumque solveris super terram erit solutum et in caelis (Mt 16,19) 196, 30[21]r (ca)]ventes in secretis pariunt et occultant. Feminae onagrorum 171, 1ra (certa)]minis tendimus. Scrinium cum reliquiis portamus contra daemones 483, 36r ] cognitio rei nisi per speciem eius sed species eventus futuri non potest 86, 25ra ] consiliis cinctus odiis 327, 11r ] contemnens suam velit statuere iustitiae domini non sit subiectus 282, 1r (cor)]rigebatur quod in ecclesia sua nolebat facere residentiam 253, 84ra ] cur timebo. Tertio dicitur quae impiis et iustis reddenda sunt ibi 19, 1ra ]date cum omnes populi 15063, 125r ] de manu Aegyptii et interfecit eum hasta sua. Haec fecit Banaias filius 20, 1ra ] deficientem et respondent totum ad temperamentum sanitatis 440, 269ra ] Dei dicuntur quia ex praecepto Dei in servitium Dei offeruntur 483, 33r ] Deum ex toto corde et ex tota anima et proximum nostrum sicut nos. Necessaria est ita quia virtus concordiae 146, 6r ] dicitur in Deum peccare 86, 36ra ] eas in sinum Abrahae in lucem sempiternam Graduale Neocellense I, 270r ] ecclesia conficitur et capiti Christo coniungitur 483, 35r (en)]sis cedit mortique vita succedit ecce vidit Graduale Neocellense II, 98r ] episcopus Brixinensis dilecto nobis in Christo Iohanni scholastici (!) de Isnina salutem in domino. Ecclesiam parochialem sancti […]usina nostrae dioecesis ad collationem nostram pleno 53, HDS (epule)]mur in azymis sinceritatis et veritatis Graduale Neocellense I, 163r ] erat cor unum et anima una et nemo dicebat aliquid 190, 31ra ] erectis quasi orans huius exemplum 146, 13ra ] esse scilicet secundum quid aliquas praelationes a Deo non esse 86, 35ra ] esse vestimenta clericorum in quibus nihil notari possit 146, 10r ] est in multis pro nobis constituta 31, 91ra ] Et merito quidem contra eos cum quadam indignatione loquitur qui in Christum dominum 190, 1ra ] et non aedificabis eos. Isti exterminio digni sunt 190, 7ra ] et probatur extra de regula veniens prope prin. Sequitur de manifesto videre 7, 64ra ] etiam Benedictus in eadem significatione 483, 3r ] exercitus in militiam congregatur. Signa militibus per turbas (!) dantur 483, 1r ] facias auxilium tuum a me ad defensionem meam Graduale Neocellense I, 130r ] faciem terrae. Alleluia Graduale Neocellense I, 207r ] faciunt sic victos e contrario voti impotes qui non possunt facere quod volunt 173, 1ra ] facultas et manumittentis 107, 54ra ] gaudet purpureosque iacit – ] quod non quia tantum consequitur 459 ] gaudet purpureosque iacit flores 173, 85ra ] Hic Deus meus et honorabo eum Deus patris mei Graduale Neocellense I, 152r (hu)]militatis gratia Petri petens vestigia 194, 22r ]ia. Benedictus es domine Deus patrum nostrorum Graduale Neocellense I, 219r ] Iam mare transcurri 327, 19r ] In serie dicta 327, 14r ] in populo Dei 15063, 84r ] indicas Hermannum VII q. I Quamvis triste et C. de codicillis nec codicillos quod vero supra dictum est 253, 85ra ] infinitum utique (!) in longitudine est et in tempore 163, 26r ] Item agitur scilicet qui […] non est generatione […] non est 405, VDS ]ituri sunt qui semper caelestia dilexerunt 190, 15ra ]ma. sed etsi § quocumque quomodo intra diem illam poterit probare laesionem 253, 83ra ] magis ad vindictam pertinet quam ad disciplinam. Caveant ergo fratres ut neminem reprehendant 146, 11r ] malum quia liberum arbitrium per se in bonum ordinatum est 86, 39ra (man)]datis tuis quae dilexi Graduale Neocellense II, 64r ] manus baptizandus et perfert fraudes temptatoris fugit persequentum lapides 154, 1r (ma)]nifestam non quae iudici nec quae alii 107, 81va ] mihi ianuam vitae 15063, 15r ] miracula et in hoc apparebit ipsorum conscientia unde Gregorius 153, 1ra ] Missa legatio dicitur Totum tempus quo Christus mundum intravit 483, 22r (ne)]mo propheta acceptus est in patria sua 171, 16ra ] non erit. Sed illi soli dicant qui dominum timent 190, 39ra ] non omnino sunt certae pronuntiationes neque vitae neque mortis. Curatio rectificetur aer 123, 11ar ] non procedit ab ore vestro sed si quis bonus est ad aedificationem fidei ut det gratiam audientibus. Ergo videtur quod etiam alii quam Christo competat esse ecclesiae caput. Propterea Christus ex eo quod praeest ecclesiae non solum dicit caput 38, 1ra ] noster in universa terra iudicia eius 617, VDS ] nugis et ioco ac fabulis vacantes. Nota ad honorem individuae trinitatis 606, 1ra (ob)]dormiam in morte ne quando dicat inimicus meus Graduale Neocellense I, 87r ] panem invenit sed cum vix ad unius prandii esum tribus personis sufficere videntur Laurentius artifices suos nolens deficere 308, 1ra ] parenti veteri tu membris fortioribus Graduale Neocellense I, 324r (Item quod est frater …)] paternam pietatem non agnoscat … Quid si aliquis interfecit patrem vel tutorem meum 7, 1ra ] peccati superbia. Prima est enim in peccando ultima remanet in conflictu 146, 8r (perma)]nens et materia transiens. Permanens est ut in cultello ferrum transiens est ut in pane farina et in vitro 605, 3ra ] per quem videmus signa in populo 440, urspr. VDS (heute abgelöst) ] poetis plerumque conceditur 173, 136ra (prae)]clarissima Amen Graduale Neocellense II, 145r ] praeformet capitis (!) naturae 327, 8r ] Prima est certitudo 146, 122ra (prin)]cipis Pharisaeorum sabbato manducare 15063, 131r ] pullorum evolantium imbecilles 171, 12ra ] quae vulgo dicitur Rothinholus. Horatius in sermone. Licinias (!) soliti impensa (!) prandere coemptas. Lucinium -nii ubi solet candela lucere quia dat lumen et dicitur a luce 106, 1ra ] quaedam forma substantialis ut anima est substantialis forma hominis alia autem et forma accidentalis ut album homini. Prima harum 605, 2ra ] quam commodum oboedientis attenderet. Itaque ut homo super hanc vitam 485, 7r ] quasi torrens praegravemur negotiis curis excitemur innumeris cogitationibus 99, 1ra ] Qui cecidit seseque potens 327, 15r ] quod non quia tantum consequitur per directam 7, 5ra 460 ] redeundum est ad mansuetudinis – ] volentes libro sexto ] redeundum est ad mansuetudinis lenitatem. Nam pertinax motus facile 146, 12r (revela)]tionem plurimum addendum fuit. Modum vero divinae cognitionis quam primus ille homo habuisse creditur explicare difficile est excepto eo quod diximus 485, 1r ] rubrum id est per baptismum vel per desertum id est paenitentiam 483, 37r ] salvos faciet eos. Hoc est enim eorum voluntas 190, 63ra ] secundo an excuset peccatum. Ad primum sic proceditur 86, 38ra ] sibi adiudicate ut extra de sententia et re iu. c. Bertoldus in fi. Si autem personali 253, 84va (si)]cut dixit alleluia 15063, 96r ] simul citant cui oboedire teneatur pro ordinario facit quia eius iurisdictio firmior et antiquior cui specialis non derogat 7, 144ra ] so sy ye meyner bedurffen, ye saliger sy sind. Noch so sol der zarten speis girikait sy nicht uben 146, 2r ] subiugat. Oppressos a diabolo liberans 483, 13r ] sumit significans angelum qui Christo resurgente lapidem 483, 19r ] tabernaculorum celebratio ordinati enim 483, 64r ]tuum montem Sion. Onus Aegypti. Ecce dominus ascendet super nubem levem 315, 1r ] tuus dolentes quaerebamus te et quid est quod me quaerebatis Graduale Neocellense I, 43r ]um filios Israel et Ioseph Graduale Neocellense I, 54r ] usitata haec enim catholicos et in schismaticos et fideles in haereticos et doctores in adulatores convertit sicut recenter nobis occurrit et nuper exprimato vidistis. Haec enim facultas est illa ancilla Agar de qua scribitur 200, 1ra ]usurarum excommunicatum publice nominatum interdictum et tempus interdicti numquid una vel quadruplici excommunicatione ligabitur 99, 1ra ] verbere et castigatione in ipso initio peccati 146,12ar (vir)]gine nascitur et per multas passiones in candorem resurrectionis 483, 21r ] virum de plebe et claritatem 15063, 65r ] Vocum quae flecti nequeunt 327, 16r ] volentes libro sexto salvis quae dicuntur in sequenti 145, 101va 461 VERFASSER UND WERKTITEL (Autoren erscheinen in der Regel unter ihrem Vornamen) Adam de Aldersbach OCist (gest. ca. 1260): Summula pauperum. Mit Kommentar und Interlinearglossen 167, 1ra Ps.-Aegidius Romanus: Speculum peccatoris s. Ps.-Augustinus Alanus ab Insulis OCist (frühscholastischer Theologe, gest. 1203): Summa de arte praedicatoria 181 Albertus de Brixia OP (gest. ca. 1314): Summa de officio sacerdotis 447, 1ra Ps.-Albertus Magnus: Secreta mulierum. Mit Kommentar 155, 123r Aldobrandinus de Cavalcantibus OP (Prediger, Bischof von Orvieto, gest. 1279): Sermo in dedicatione ecclesiae 162, 8ra Aldobrandinus de Toscanella (Lektor in Pisa, Pistoia, Siena, Viterbo 1287–1292): Scala fidei 31, 1ra Alexander de Villa Dei (Grammatiker, Mathematiker, gest. ca. 1250): Doctrinale (Fragmente) 52, Fälze Alfonsus Bonihominis OP (Arabist, gest. 1339) s. Rabbi Samuel Ambrosius Spiera OSM (ca. 1413–1454/1455): Sermo Fer. 6 in parasceve 174, 156Ira Anrufung an den hl. Hartmann 293, 23v Antiphonarium 15063; 139, 154r; 940, 152r Antonius Guainerius (Arzt aus Pavia, gest. nach 1445): – Summarium de febribus 748, 1v – Tractatus de peste et venenis 748, 90v Antonius Volscus: Vita Ovidii 579, 1r (am Rand) De aqua vitae 748, 53ra Argumenta librorum Dictys Cretensis: De Troiani regni excidio cum appendiculis 691 Aristoteles (Philosoph, gest. 322): – De anima, translatio nova durch Guilelmus de Moerbeke. Mit Kommentar 163, 32r – Physica, Lib. 6–8, Translatio nova durch Guilelmus de Moerbeke. Mit Kommentar 163, 1r (Forts. von Cod. 168, 46v) – Physica, Lib. 1–5, Translatio nova durch Guilemus de Moerbeke. Mit Kommentar 168, 1r – s. auch Iohannes Buridanus – s. auch Quaestiones secundum Iohannem Buridanum super libros Ethicorum Aristotelis I–V Arnoldus de Geilhoven CRSA (gest. 1442): Remissorium utriusque iuris (Praefatio) 4, 1ra Ars notariatus 155, 115ra Ars praedicandi 155, 139r Astronomische Texte 776, 47r Auctoritates (Zitate): – zu De Anima 597, 57vb – philosophische 155, 144ra – verschiedene 308, 225va; 755, 10ra Augustinus, hl. (Kirchenvater, gest. 430): – Confessiones 158, 1ra – Exzerpte 167, 71va – Sermo 9: De decem chordis 165, 141r – s. auch Regula s. Augustini Ps.-Augustinus: – Auctoritates (Exzerpte) 165, 42ra – Epistola 18 (ad Cyrillum Hierosolymitanum episcopum de magnificentiis beati Hieronymi) 125, 183vb; 165, 53va – De praedestinatione Dei 532, 27r – Sermo 277: De reddendis decimis 155, 16ra – Speculum peccatoris 83, 52va 462 Augustinus Favaroni – Ps.-Cyrillus Augustinus Favaroni OESA (Generalprior, Erzbischof von Nazareth, 1360–1443): Sermones bzw. Interrogationes seu introductiones quadragesimales 597, 43va Avianus Poeta (röm. Fabeldichter, ca. 4./5. Jh. n. Chr.): Fabeln (Nr. 1–28). Mit Kommentar 155, 155v Bartholomaeus Brixiensis (Kanonist, 1200–1258): Quaestiones dominicales, De conditionibus appositis, unvollständig 614, 289v Bartholomaeus a Sancto Concordio OP (Kanonist, gest. 1347): Summa de casibus conscientiae (= Summa Pisanella) 145, 2ra Beda Venerabilis OSB (Kirchenlehrer, gest. 735): Exzerpte 167, 71va Ps.-Beda Venerabilis: Termini quartae decimae lunae regulares (Memorialverse zur Berechnung des Osterfestes) 360, 26r Beimiarm s. Isaac Iudaeus Bernardinus Senensis OFM (Prediger) (?): Speculum peccatorum sive amatorum mundi, unvollständig 83, 55va Bernardus Claraevallensis OCist, hl. (Kirchenlehrer, gest. 1153): – De gradibus humilitatis et superbiae 282, 1r – De gratia et libero arbitrio, unvollständig 282, 1r – Epistola 17 (ad Henricum Senonensem archiepiscopum: De moribus et officio episcoporum) 282, 15r – s. auch Tractatus super missus est Ps.-Bernardus Claraevallensis: Speculum peccatoris s. Ps.-Augustinus Bernardus de Gordonio (Arzt, ca. 1283–1320): Practica seu Lilium medicinae, unvollständig 123, 1r, 215v (Nachtrag), 218r (Nachtrag) Bernardus Papiensis (Bischof von Faenza und Pavia, gest. 1213): Breviarium Extravagantium sive Compilatio prima Decretalium, unvollständig 107, 1ra Bernardus Parmensis de Botone (Kanonist, gest. 1266): Casus longi Decretalium (Exzerpte) 155, 106ara Bibel s. Vetus testamentum Bonaventura OFM (Kirchenlehrer, gest. 1247) (Bonaventura abbreviatus): Super II Sententiarum 52, 43v Ps.-Bonaventura: – Stimulus amoris s. auch Iacobus Mediolanensis – Tabula a celebrantibus missam consideranda 167, 72r Bonifatius VIII. (Papst 1294–1303): Regulae iuris 614, 282v Breviarium Salisburgense 405 Bruno de Segni OSB (Abt von Montecassino, Bischof von Segni, ca. 1048–1123): Expositio in Psalmos, unvollständig 190, 1ra Caesarius Heisterbacensis OCist (Mönch, 1180–1240): Dialogus miraculorum, Dist. 3, cap. 6 125, 284ra; 308, 126vb Caesarius Arelatensis, hl. (Bischof von Arles, Kirchenschriftsteller, 469/70–542): Sermo 33 155, 16ra Canones: Begriffserklärungen (wohl Kompilation aus mehreren Rechtslexika) 155, 18ra Cedula (Abschrift), Augustiner Chorherrenstift Neustift 76, 71Ir Chalmelli, Raimundus (Arzt, ca. 1334–1398): De pestilentia libri tres, unvollständig 748, 54v Christophorus Barsisius (Professor der Medizin in Padua 1434–1440): De febrium cognitione et cura 748, 41r Cicero, Marcus Tullius (röm. Politiker, Anwalt und Philosoph, 106–43 v. Chr.): De officiis 1,22 755, 10rb Claustrum animae 165, 144rb Clemens V. (Papst 1305–1314): Constitutiones (Clementinen), unvollständig 99, 1ra De comparationibus BMV 171, 62ra Conradus de Brundelsheim (Soccus) OCist (Abt von Heilsbronn, gest. 1321): Sermo in dedicatione ecclesiae 162, 5rb Conradus de Waldhausen OSACan (Prediger, gest. 1369): – Flores (Graeculus Piper) secundum Conradum de Waldhausen 177, 1ra – s. auch Graeculus Conradus Eberbacensis OCist (Kirchenschriftsteller, Abt von Eberbach, gest. 1221): – Exordium magnum Cisterciense, Dist. 4 (Auszüge) 125, 261ra, 272rb; 308, 76va, 101rb – Exordium magnum Cisterciense, Dist. 5 (Auszüge) 125, 272va; 308, 101va, 101vb Conradus Hirsaugiensis OSB (Mönch in Hirsau, Philosoph, Schriftsteller, Musiker, ca.1070–1150): Dialogus super auctores, unvollständig 360, 1r Constantinus Africanus (Arzt, um 1010–1057): – Praefatio 748, 162ra – s. auch Isaac Iudaeus Ps.-Cyrillus: Epistola ad Augustinum de miraculis Hieronymi 125, 186va; 165, 55vb De … (anonyme Texte) – Godefridus de Trano 463 De … (anonyme Texte) s. unter dem ersten Substantiv Declaratio arboris consanguinitatis 155, 105v Determinatio doctorum universitatis Pragensis de V propositionibus magistri Iohannis Muntzinger 1386 facta = Gutachten der Prager theologischen Fakultät im Rahmen der theologischen Streitigkeiten zw. Johannes Müntzinger und den Dominikanern in Ulm 165, 129r Dionysius Carthusianus (Theologe, „Doctor ecstaticus“, 1402–1471): Revelatio cuiusdam Carthusiensis in missa 754, 11v Dispositio catholicae fidei, unvollständig 190, 67rb Divisiones libri tegni Galeni 453, 33v Donatus, Aelius (Grammatiker, Mitte 4. Jh.) s. Terentius Afer, Publius Donatus devotionis: De octo partibus orationis, Pars 1 617, 1r Durantis, Guilelmus (Kanonist, gest. 1296): – Rationale divinorum officiorum 550, 96ra – Speculum iudiciale, unvollständig 7, 1ra Eadmerus Cantuariensis OSB (Bischof von St. Andrews, gest. 1124): De beatitudine coelestis patriae 532, 28v Ebendorfer, Thomas (Theologe, Rektor der Universität Wien, 1388–1464): Sermones de tempore de evangeliis, pars hiemalis 53, 1r Ecbertus Schonaugiensis OSB (Theologe, ca. 1120–1184): Soliloquium seu Meditationes 125, 169va (Auszug); 165, 139v Engelschalk, Albertus (Theologe, Rektor der Universität Prag und Wien, ca. 1353–1430) s. Ambrosius Spiera Engelschalk, Matthias (Magister in Niederaltaich, 15. Jh.): De passione Christi secundum quattuor evangelistas 756 Epistola de visione cuiusdam fratris ordinis Carthusiensis in Ruremunda 754, 13r Eusebius Caesariensis (Bischof von Caesarea, griechischer Kirchenschriftsteller, gest. 339): Ecclesiasticae Historiae, ins Lat. übersetzt von Rufinus Aquileiensis (Auszüge) 166, 52va Ps.-Eusebius Caesariensis: Epistola ad beatum Damasum episcopum Portuensem et ad christianissimum Theodosium Romanorum senatorem de morte Hieronymi 125, 170rb; 165, 42va; 182, 1ra Exempla s. auch Miracula Expositio orationis dominicae 161, 8va Expositio super Ave Maria 31, 49ra Flavius Iosephus (jüdischer Historiker, gest. ca. 100): – De antiquitatibus (Auszüge) 52, 38vb – De bello Iudaico (Auszüge) 52, 38vb – De forma vivendi monachorum 165, 143va Fragmente: – philosophischer Traktat 405, VDS – Psalterium 617, VDS – Urkunde des Bischofs von Brixen an Johannes, scholasticus aus Isny (de Ysnina) 53, HDS Franciscus de Zabarellis (Theologe, 1339–1417): De horis canonicis 155, 1ra Galenus (griechischer Arzt und Philosoph, ca. 130–200): – Ars parva seu Microtegni, ins Lat. übersetzt von Gerardus Cremonensis 453, 1r – s. auch Divisiones libri tegni Galeni Galfredus de Vinosalvo (Grammatiker, gest. um 1210): Poetria nova, unvollständig 327, 1r Gentilis de Fulgineo (Arzt, Naturphilosoph, gest. 1348): Consilium de peste, Textbeginn 748, 66v Gerardus Cremonensis (Übersetzer aus dem Arabischen und Griechischen ins Lateinische, gest. 1187) s. Galenus Gerardus de Vliederhoven OT (Theologe, 2. H. 14. Jh.): Cordiale quattuor novissimorum 125, 125va Gerson, Iohannes (Theologe, gest. 1429): – De modo confessionis et absolutionis sacramentalis 755, 8rb – De modo excommunicationum 755, 8vb – De oratione et suo valore 754, 1r – De probatione spirituum 755, 1ra Gerson, Iohannes (Theologe, gest. 1429) (?): De iniunctione paenitentiae 755, 9va Godefridus Altissiodorensis (franz. Zisterzienserabt, Mönch in Clairvaux, ca. 1115/20–1188): Declamationes ex s. Bernardi sermonibus, cap. 25 125, 311rb; 308, 187va Godefridus Herilacensis OSB (Theologe, 15. Jh.) (= Ps.-Iacobus de Paradiso): Expositio super orationem dominicam, unvollständig 31, 92ra Godefridus de Trano (Jurist, Kardinaldiakon, gest. ca. 1245): Summa super rubricis Decretalium 253, 1ra 464 Graduale – Iacobus Forliviensis Graduale 139, 1r; 940, Ir Graduale Neocellense (2 Bde) (ohne Sign.) Graeculus (Titelname von lateinischen Predigten um 1300): – Sermo in annuntiatione BMV 617, 148r – s. auch Conradus de Waldhausen Grammatica speculativa s. auch De modis significandi Gregorius Magnus, hl. (Kirchenlehrer, Papst 590–604): – Dialogi 83, 1ra; 165, 1ra; 495 – Dialogi, Lib. 1 (Auszüge) 125, 307ra; 308, 177rb – Dialogi, Lib. 3 (Auszüge) 308, 198va, 200ra – Dialogi, Lib. 4 (Auszüge) 125, 275va, 302ra, 305rb; 308, 107va, 166rb, 173ra, 199va – Exzerpte 167, 71va – XL Homiliae in Evangelia (Auszüge) 125, 288Iva; 308, 139va Ps.-Gregorius Magnus: Dialogi, Lib. 2 (Auszüge) 125, 246vb; 308, 55vb Grimlaicus (um 900): Regula solitariorum (Auszüge) 125, 280rb, 300va, 301rb, 303va, 310vb; 308, 118ra, 163rb, 164va, 169ra, 186ra Guarinus Veronensis (Humanist, Übersetzer, 1374–1460): Argumenta Iuvenalis Saturarum 217, 1r Guido de Monte Rocherii (Pfarrer von Teruel in Aragonien, 14. Jh.): Manipulus curatorum 550, 1ra Guilelmus de Moerbeke OP (Bischof von Korinth, Übersetzer aus dem Arabischen und Griechischen ins Lateinische, gest. 1286) – Aristoteles: De anima, translatio nova 163, 32r – Aristoteles: Physica, translatio nova 163, 1r (Lib. 6–8; Forts. von Cod. 168, 46v); 168, 1r (Lib. 1–5) Guilelmus Malmesburiensis OSB (Historiograph, Bibliothekar, 1080–1142): Gesta regum Anglorum (Auszüge) 125, 213va, 272vb, 282va, 287ra, 287rb; 308, 102ra, 123ra, 133vb, 134va, 136rb Guilelmus Peraldus OP (Prediger, gest. ca. 1271): Summa de vitiis et virtutibus (Exzerpte) 125, 98ra Henricus Suso (Mystiker, gest. 1366): Horologium sapientiae 183 Ps.-Henricus de Frimaria: In libros I–IV Sententiarum sive Filia magistri 532, 34v Herbertus Claraevallensis OCist. (ca. 1168/98): – De miraculis, Lib. 1 (Auszüge) 125, 279va; 308, 116ra – De miraculis, Lib. 2 (Auszüge) 125, 250rb, 250va, 272rb, 272va; 308, 60vb, 61rb, 101rb, 101va, 101vb – De miraculis, Lib. 3 (Auszüge) 125, 216va, 277va, 279vb, 282vb, 284vb; 308, 111va, 116vb, 123vb, 128rb Herolt, Iohannes OP (Prior und Lektor in Nürnberg, gest. 1468) (Discipulus): Sermones de tempore 153, 1ra Hieronymus, hl. (Kirchenvater, gest. 420): – Bibelprologe 20; 204 – Epistola ad Helvidium (Auszüge) 52, 38vb – Epistola 107 (ad Laetam) (Auszug) 125, 303va; 308, 169rb Ps.-Hieronymus: Speculum peccatoris s. Ps.-Augustinus Hippocrates (griechischer Arzt, 460–370 v. Chr.): – Aphorismi 453, 39r – s. auch Quaestiones in Hippocratis Aphorismos Historia passionis dni secundum quattuor evangelistas 174, 156Ira Honorius Augustodunensis (Scholastiker, 12. Jh.): – Gemma animae, unvollständig 483 – Speculum ecclesiae (Auszüge) 125, 271Iva, 288Ira; 308, 99vb, 138va Horatius Flaccus, Quintus (römischer Dichter, 65–8 v. Chr.): Auszüge aus u. a. Ars poetica, Carmina, Epistolae, Sermones 755, 11va Hugo de Sancto Caro OP (Theologe, Kardinal, gest. 1263): Postilla in Proverbia, unvollständig 156 Ps.-Hugo de Sancto Caro: In libros I–IV Sententiarum sive Filia magistri s. Ps.-Henricus de Frimaria Hugo de Sancto Victore CRSA (Scholastiker, Mystiker, gest. 1141): – De orando Deum 755, 4ra – De sacramentis, Lib. 1, unvollständig 485 Hugo de Sancto Victore (?): Expositio in regulam beati Augustini, unvollständig 142, 7r; 146, 6r Humbertus de Prulliaco OCist (Abt von Preuilly, gest. 1298): Extractiones super IV librum Sententiarum Petri Lombardi 52, 110v Humbertus de Romanis OP (Prior in Lyon, gest. 1277): Expositio regulae s. Augustini, unvollständig 146, 13ra Iacobus Forliviensis (Arzt, Lektor an der Universität Padua, ca. 1360–1414): – Expositio in Hippocratis Aphorismos 453, 59ra – Quaestiones primi tegni Galeni 453, 187ra Iacobus de Lausanna OP – Kanonistische Kurztexte 465 Iacobus de Lausanna OP (Prediger, gest. vor 1332): Compendium moralitatum 52, 202ra Iacobus Mediolanensis OFM (Theologe, Ende 13. Jh.) = Ps.-Bonaventura: Stimulus amoris (Auszug) 125, 161ra; 165, 132v Ps.-Iacobus de Paradiso s. Godefridus Herilacensis Iacobus de Voragine OP (Prediger, Hagiograph, gest. 1298): Legenda aurea sanctorum, sive Lombardica historia, cap. 35: De ieiunio quattuor temporum 52, 39rb Innocentius III. (Papst 1198–1216): – De contemptu mundi, Lib. 2 (Auszüge) 125, 306ra; 308, 174va – De contemptu mundi, Lib. 3 (Auszüge) 125, 271Ira; 308, 98vb Iohannes XXI. (Papst 1176–1177) s. Petrus Hispanus Iohannes XXII. (Papst 955–963): Extravagantes, unvollständig 99, 33vb Iohannes Andreae (Kanonist, gest. 1348): – Hieronymianus (Auszüge) 125, 198va; 165, 71rb – Novella super librum sextum Decretalium, unvollständig 7, 144ra – Summula quarti libri Decretalium (Summa de sponsalibus et matrimonio) = Überarbeitung von Iohannes de – Anguissola: Summa de sponsalibus et matrimoniis 99, 32va – s. auch Clemens V. – s. auch Summarium Quaestionum mercurialium Ps.-Iohannes Andreae: Ordo iudiciarius 155, 118rb Iohannes de Anguissola s. Iohannes Andreae Iohannes de Auerbach (Domvikar in Bamberg, Kanonist, 15. Jh.): Processus iudiciarius 155, 51ra Iohannes Buridanus (Rektor in Paris, ca. 1300–1358): – Quaestiones super libros Meteororum Aristotelis, unvollständig 440, 269ra – s. auch Quaestiones secundum Iohannem Buridanum super libros Ethicorum Aristotelis I–V Iohannes Calderinus (Kanonist, ca. 1300‑1365): – Repertorium utriusque iuris 4, 1ra – s. auch Arnoldus de Geilhoven Iohannes Calphurnius (Humanist, Professor in Padua, gest. 1503) s. Terentius Afer, Publius Iohannes Cassianus (frühchristlicher Schriftsteller, ca. 360–435): Vitae patrum, Lib. 4 (Auszüge) 125, 309ra, 309vb; 165, 104vb; 166, 57ra; 308, 181vb, 183vb Iohannes Contractus (Korz) OFM (Prediger, 2. H. 14. Jh.): Sermo de s. Andrea 617, 194r Iohannes de Dambach OP (Theologe, gest. 1372): Tractatus de indulgentiis 161, 1ra Iohannes de Deo (Kanonist, 13. Jh.): Concordantiae Decretalium et Decretorum 614, 270r Iohannes Friburgensis OP (Theologe, gest. 1314): Confessionale 532, 125r Iohannes Guallensis (Wallensis) OFM (Kanonist, gest. 1285 in Paris): Compilatio secunda Decretalium, unvollständig 107, 79ra Iohannes Ianuensis OP (Theologe, Lexikograph, gest. 1298): Catholicon sive Prosodia, Pars 5: Vocabularium, unvollständig 106 Iohannes Marchesinus OFM (Theologe, gest. vor 1312): Super Psalterio (aus: Mammotrectus super Bibliam) 155, 141r Iohannes Mesue: Liber aphorismorum 453, 35r – s. auch Synonyma Rasis Iohannes de Sancta Sophia (Arzt aus Padua, 2. H. 14. Jh.): Consilium de pestilentia 748, 71r Iohannes de Tornamira (Arzt in Montpellier, gest. vor 1396): Preservatio et cura apostematum antrosorum pestilentialium 748, 69v Iohannes de Turrecremata (Theologe, 1388–1468): Meditationes 397, 1r Isaac Iudaeus (= Beimiarm) (Arzt und Philosoph, gest. um 955), ins Lat. übersetzt von Constantinus Africanus (Arzt, um 1018–1087): – Liber de febribus 748, 162ra – Liber de urinis 748, 138ra Isaias mit Glossa ordinaria, unvollständig 315 Isidorus Hispalensis, hl. (Kirchenlehrer, Bischof von Sevilla, gest. 636): De summo bono seu Sententiarum Lib. 1–3 (Exzerpte) 52, 2va Iuvenalis, Decimus Iunius (röm. Satiriker, ca. 60/90–140): Saturae 217, 1r, 45r (jeweils unvollständig); 755, 11va, 12ra (jeweils Auszüge) Kempf, Nicolaus OCart (Theologe, Prior von Geirach, Gaming und Pleterje, gest. 1497): Tractatus de proponentibus religionis ingressum et de anno probationis 617, 63r Kanonistische Kurztexte 145, 314va; 155, 6va, 15ra, 16vb, 48ra, 49vb; 614, 273v 466 Lacteus liquor – Notizen Lacteus liquor, unvollständig 308, 1ra Lectionarium missae Neocellense 196 Leo OCist. (Prediger, 14. Jh.): Speculum corporis Christi sive Dialogus increduli et ecclesiae de sacramento eucharistiae 617, 150r Libellus officiorum 76, 53v Liber anniversariorum Neocellensis 142, 52ra Liber Horarum 353 (Bd. 1); 654 (Bd. 2) Liber miraculorum dictus Lacteus s. Sibylla Tiburtina Liber sermonum „Fons vitae“ 174, 169rb Marsilius de Inghen (Scholastiker, Magister an der Universität Paris und Heidelberg, ca. 1340–1396): Quaestiones parvorum naturalium Aristotelis 440, 1ra Martinus V. (Papst 1417–1431): Bulla „Regimini universalis“ 447, Xr Martinus Oppaviensis OP (Theologe, gest. 1278): Sermo in dedicatione ecclesiae 162, 9vb Medizinische Rezepte, Anweisungen, Abhandlungen und Notizen: 453, 53va; 748, Ir, 1r, 40v, 52r, 52v, 54ra, 64v, 66r, 69v, 70v, 72r, 212v, 218vc – zur Heilung von Pferdekrankheiten 160, 1r Memorialverse: – De ordine librorum Bibliae 166, IIv – Berechnung des Osterfestes s. Ps.-Beda Venerabilis Miracula: – Miraculum de duobus sociis 125, 311v – verschiedene: 308, 187vb, 201va – s. auch Lacteus liquor – s. auch Sibylla Tiburtina Missale: – Missale (festivum) 100 – Missale speciale (Teilmissale) 154, 1r – De modis significandi (Grammatica speculativa), unvollständig 605, 5va Mundinus de Leuciis (Arzt, ca. 1270–1326): Anatomia 748, 74r Müntzinger, Iohannes (Theologe, Rektor der Stadtschulen in Ulm um Rottweil, gest. 1417) s. Quaestio de articulis und s. Determinatio doctorum Musicae modulationis commendatio 776, 1r Musiktraktat 776, 13r Necrologium Neocellense 142, 127ra Neustift, Augustinerchorherrenstift: – Kapitelbeschluss von 1653, betreffend die Vigil des Augustinustages in Neustift und die Frage der Abstinenz 76, 74r – Über die Pfründnerschaft im Kloster, dt. 76, 73r – Verfügung des Propstes Fortunat Troyer (1678–1707), betreffend die Offizien beim Begräbnis eines Klerikers 726, 74r – Vermerke über den 1581 verstorbenen Neustifter Propst Augustinus Schbäl (recte: Schabl) und den 1581 zum Propst gewählten Adamus Lang 726, 76r – Visitationsinstrument (Abschrift), Augustiner Chorherrenstift Neustift 76, 66r Ps.-Nicolaus de Dinkelsbühl: Speculum artis bene moriendi 754, 18r Ps.-Nicolaus de Gorran: Postilla in Proverbia s. Hugo de Sancto Caro Nicolaus Pergamenus (Fabeldichter, 15. Jh.): Dialogus creaturarum (Auszüge) 125, 304va; 308, 171va Notizen: – zu De anima 163, 79v – zu De baptismo 447, IXra – aus Kloster Baumburg (Oberbayern) über die Weihe der dortigen Kirche 293, 22r – komputistische 617, 195r – zu Unterschieden zw. der griech. und der lat. Kirche 52, VDS – verschiedene 52, VDS; 165, 111rb; 167, 72r; 173, 172r; 360, 26r; 397, VDS; 453, VDS; 532, 124v – zur Verwandtschaft der hl. Maria 293, 23r – zur Verwendung der Alhidade mit Skizze 776, 71v – zur Weihe der Kirche zu Seckau 293, 22r Odo de Cheriton – Robertus Anglicus 467 Odo de Cheriton (englischer Theologe, gest. ca. 1246): Parabolae (Auszüge) 125, 282ra, 297rb; 308, 121va, 156va Ordenshandschriften: – s. Tabelle mit Auflistung der Ordensgemeinschaften – s. Neustift, Augustiner Chorherrenstift Ordinarium missae 252 De ordine librorum Bibliae s. Memorialverse Ordo baptisterii 726, 71r Ps.-Origenes: Homilia de s. Maria Magdalena 161, 6ra Ovidius Naso, Publius (röm. Dichter, 43 v. Chr.–17 n. Chr.): – Ars amatoria (Auszug) 755, 11vb – Remedia amoris. Mit zahlreichen Interlinear- und Marginalglossen 579 – s. auch Antonius Volscus Peregrinus Oppoliensis OP (Prior in Breslau und Ratibor, gest. um 1322): Sermones de sanctis 78, 1ra Persius Flaccus, Aulus (röm. Dichter, 34–62): Saturae, unvollständig 217, 35r Pestregimen 748, 71r, 72r Petrus Beneventanus Collivacinus (Kanonist, päpstlicher Notar, gest. 1219/20): Compilatio tertia Decretalium, unvollständig 107, 81ra Petrus Blesensis (Theologe, 1135–1212): Compendium in Iob 77, 113ra Petrus Cantor (Magister und Kanonikus an der Kathedralsschule von Notre-Dame in Paris, ca. 1130–1197): Verbum abbreviatum (Auszüge) 125, 304va, 307ra; 308, 171rb, 177ra Petrus Hispanus (Philosoph, ab 1276 Papst Iohannes XXI., ca. 1210–1277): Tractatus sive Summulae logicales, unvollständig 605, 1ra Petrus Lombardus (Scholastiker, Bischof von Paris, um 1095–1160): – Commentarii in Psalmos, unvollständig 19 – s. auch Richardus de Mediavilla De plebanis 76, 63v Pomponius Laetus, Iulius (Humanist, 1428–1498): Widmungsbrief zur Sallustedition an Augustinus Maphaeus 780, 1r Practica contra pestem 748, 66v Processus iudiciarius 614, 150r – s. auch Iohannes de Auerbach Prosdocimus de Beldemandis (Mathematiker, Astronom, Musiktheoretiker, gest. 1428): Compositio astrolabii 776, 61r Ps(eudo-) s. nach dem Namen des jeweiligen Autors Quadragesimale De schola Christi (48 Predigten) 597, 1ra Quaestio de articulis Iohannis Müntzinger 165, 131v Quaestio sollemnis 76, 11r Quaestiones disputatae de peccato in spiritum sanctum 165, 114r Quaestiones in Hippocratis Aphorismos, unvollständig 453, 143ra Quaestiones secundum Iohannem Buridanum super libros Ethicorum Aristotelis I–V, unvollständig 440, 89ra Rabbi Samuel (11. Jh.): Epistola ad Rabbi Isaac, ins Lat. übersetzt von Alfonsus Bonihominis 617, 117r Raber, Vigil: Neustifter Wappenbuch ohne Sign. Radbertus Paschasius OSB (Abt von Corbey, ca. 788–858): De corpore et sanguine dni, cap. 14 125, 278vb; 308, 114rb Raimundus de Pennaforte OP (Professor des Kirchenrechts in Bologna, gest. 1275): – Summa de matrimonio (= Summa, Lib. 4) 253, 85ra (unvollständig); 614, 139r – Summa de paenitentia (= Summa, Lib. 1–3) 614, 1r Rampegolus, Antonius OESA (ca. 1360–1423): Biblia aurea 125, 1ra De reemptionibus 447, 208ra Regimen Sanitatis Salernitanum 748, 1r (sechs Verse), 176ra (vier Verse), 212vb (sechs Verse) Regula s. Augustini 76, 1r; 142, 1r; 146, 1r (dt., unvollständig) Regulae iuris civilis (alphabetisch geordnete Sammlung von Rechtssprichwörtern und -regeln [A–U/V]) 614, 275r Richardus de Mediavilla OFM (Theologe, 1249–1302): Commentarius in Petri Lombardi Sententiarum librum secundum, Prologus 532, 123r Rituale Neocellense 194; 726; 14820 Robertus Anglicus (Mathematiker in Montpellier, um 1250/71) (?): De astrolabio 776, 47r 468 Robertus Holcot – Thomas de Aquino Robertus Holcot OP (Theologe, Lehrer in Cambridge und Oxford, gest. 1349): – Commentarius in librum Sapientiae, cap. 3 (Exzerpte) 617, 196r – Postilla super librum sapientiae, unvollständig 200 Rufinus Aquileiensis (frühchristlicher Schriftsteller, ca. 435–510): – Historia monachorum (= Vitae patrum, Lib. 2) (Auszüge) 125, 305va, 307rb, 309ra, 309va; 165, 75ra; 308, 173va, 177va 182ra – s. auch Eusebius Caesariensis Ps.-Rufinus Aquileiensis: Vitae patrum, Lib. 3 = Verba seniorum (Auszüge) 125, 303va, 305ra, 305vb, 307rb, 307va, 308vb, 311rb; 165, 88va; 308, 169ra, 172va, 174rb, 178ra, 187ra Rusius, Laurentius (Veterinär, 14. Jh.): Marescalcia 160, 2ra Sallustius Crispus, Gaius (röm. Historiograph, Politiker, 86–35 v. Ch.): – De bello Iugurthino 780, 37r – De coniuratione Catilinae 755, 11vb (Auszug); 780, 2r De schola Christi s. Quadragesimale De schola Christi Serapion s. Synonyma Rasis Serlon de Wilton (englischer Dichter, ca. 1110–1181): Versus 158, IIr Sermones, anonym: – Sermo 597, 58ra (unvollständig); 78, 112ra – Sermones in dedicatione ecclesiae 162, 1ra – Sermones de epistolis de tempore „Sensatus“. Mit Konkordanz I und II 174, 1ra – Sermones de sanctis et de tempore 617, 134v – Sermones de tempore 174, 155ra – s. Liber sermonum „Fons vitae“ Servasanctus de Faenza OFM (gest. um 1300): Sermones 617, 200r Servius, Maurus Honoratus (röm. Grammatiker, ca. 4./5. Jh.): – Commentarius in Vergilii Aeneidos, unvollständig 173, 27ra – Commentarius in Vergilii Bucolica, unvollständig, 173, 1ra – Commentarius in Vergilii Georgica 173, 3vb Sibylla Tiburtina (= Liber miraculorum dictus Lacteus) 125, 205ra Sigbertus Gemblacensis (Chronograph, ca. 1030–1112): Chronica 823 125, 281vb; 308, 121va Speculum humanae salvationis 166, 1ra Statius, Publius Papinius (röm. Epiker, 45–96): Thebais II 620 (Federprobe) 173, 172r Statuta canonicorum regularium sub episcopis Pataviensibus Georgio et Leonhardo (1389–1451) ordinata pro monasteriis BMV in Dürnstein et s. Dorothea (Wien) (sog. „Wiener Statuten”) 76, 17r Strambotto (ital.) 4, Iv Sulpicius Severus (frühchristlicher Schriftsteller, um 400): – Dialogi, Lib. 1 (Auszüge) 125, 283va, 288rb; 308, 125va, 136vb – Vitae patrum, Lib. 4 (Auszüge) 125, 309ra, 309vb; 165, 104vb; 166, 57ra; 308, 181vb, 183vb Summarium Quaestionum mercurialium Iohannis Andreae 614, 284r Synonyma Rasis 748, 213ra Tabelle mit Auflistung der verschiedenen Ordensgemeinschaften 155, 17va Tafel zur Berechnung des Osterfestes mit erläuternden Versus de termino paschae (19 Zeilen) 360, 26v Tancredus Bononiensis (Kanonist, 1185–1236): Ordo iudicarius, unvollständig 253, 83ra, 84ra Terentius Afer, Publius (röm. Dichter, 195/184–159/158 v. Ch.), mit vereinzelten Auszügen aus dem Kommentar des Iohannes Calphurnius und des Aelius Donatus 755, 10ra Termini librorum I–V Decretalium 155, 45va Theobaldus (11. Jh.): Physiologus cum commento 155, 151r Thomas a Kempis OSA (Theologe, gest. 1471): De imitatione Christi 125, 146ra Thomas Cantimpratensis OP (Theologe und Naturforscher, 1201–1263): Liber de natura rerum, unvollständig 171, 1ra Thomas de Aquino OP, hl. (Kirchenlehrer, gest. 1274): – Catena aurea in Lucam (Exzerpte) 52, 38vb – Summa theologica, pars 3. Quaestiones de incarnatione dni, unvollständig 38 – Super librum secundum Sententiarum, unvollständig 86 – s. auch Trakate Traktate – „Wiener Statuten“ 469 Traktate: – Traktat in enger Anlehnung an: Thomas de Aquino: Super librum quartum Sententiarum 77, 1ra – Musiktraktat 776, 13r – de absentis praesentia 532, 1r – de anthrace 748, 90r – de asmate 123, 212r – de confessione, unvollständig 606 – de decem praeceptis 308, 210ra – de divisionibus in musica 776, 23r – de febribus 123, 195r – de sacramentis 85, 1r – quomodo religiosus proficere possit in religione 617, 55r – super Missus est (Exzerpte aus Bernardus Claraevallensis) 171, 88rb – über die Legende der 30 Denare, unvollständig 308, 228vb – De transsubstantiatione 167, 71vb Urkunden: – Notariatsinstrumente Brixen 1420 und 1446 (jeweils Abklatsch) 550, VD, HD (urspr. Einband lose) – Urkundenvorlage (anonymisiert) 253, 83v – s. auch Fragmente Ursinus, Paulus (Arzt, 1. H. 16. Jh): Dialogus de formidine diluvii instantis anni vigesimi quarti abigenda 694 De usura 447, 207ra Verba seniorum: – s. Ps.-Rufinus Aquileiensis: Vitae patrum, Lib. 3 – s. Vitae patrum, Lib. 5 – s. Vitae patrum, Lib. 6 Vergilius Maro, Publius (röm. Dichter, 70–19 v. Chr.): Georgica (Federprobe) 173, 172r Verse: – über drei Heilige namens Joseph (34 Verse) 293, 23r – zu Stationen im Leben Mariens und Jesu 308, 228rb – de ornamentis filiarum Ierusalem 483, 76v – de regimine sacerdotis 167, 71vb – de sanctis Iohanne Baptista et Iohanne ev. 52, VDS – de termino paschae s. Tafel zur Berechnung des Osterfestes – Versus de titulis librorum Decretalium 145, 1r – s. auch Regimen Sanitatis Salernitatum – s. auch Serlon de Wilton – s. auch Strambotto – s. auch De transsubstantiatione – s. auch Vita s. Mariae Vetus testamentum 20 (unvollständig); 204 Visio Philiberti 308, 189ra Vita beati Hartmanni episcopi (Brixinensis) 293 Vita s. Mariae post ascensionem Christi versificata 166, 51va Vitae patrum: – Lib. 2 s. Rufinus Aquileiensis: Historia monachorum – Lib. 3 s. Ps.-Rufinus Aquileiensis – Lib 4 s. Iohannes Cassianus, Sulpicius Severus – Lib. 5: Verba seniorum 125, 301rb, 302rb, 302vb, 303ra, 303rb, 304rb, 306rb; 308, 164va, 166va, 167va, 168ra, 168vb, 170vb, 184va, 185ra, 185rb, 185vb, 186ra – Lib. 6: Verba seniorum 125, 305vb, 307rb; 308, 174rb, 178ra, 186rb, 186vb Vitalis Blesensis (franz. Dichter, 12. Jh.): Geta, unvollständig 327, 20ra Volgosius, Raphael: Alphabetisches Titelverzeichnis zum Römischen Recht 578, 1ra Votivmessen und Gebete für Verstorbene 754, 26v Wernerus de Ellerbach OSB (Abt von St. Blasien, gest. 1126): Deflorationes ss. patrum, Lib. 1, Sermo in rogationibus 125, 288Ira; 308, 138rb „Wiener Statuten“ s. Statuta 470 VORBESITZER, BENÜTZER, AUFTRAGGEBER UND KÄUFER Bartolomeo da Cremona 173 (Buchdrucker in Venedig) Bertholdus (Berchtoldus hospitalis), Hospitaler zu Brixen 7; 99; 253 (Besitzer, Geschenkgeber) Bertoldus de Saxonia, Präbendar und Arzt in Kloster Neustift 123, Vv; 453; 748, IIIv (Besitzer, Geschenkgeber) Ebner, Johann (Brixner Domherr, gest. 1460) 4, IIv, 234r, 431va (Besitzer) Ephensteiner, Petrus 76 (Kopist der Hs. bzw. Schreiber einer Hs. aus dem Jahr 1728) Führer, Paul Nikolaus 76 (Buchdrucker in Brixen) Geczenperger, Nikclas 940, HDS Golser, Georg (Bischof von Brixen 1464–88) 353, IVrb Härber von Ringelsberg, Lukas (Propst von Neustift 1483–1503) 405 (Auftraggeber, Besitzer) Helene 578, Ir (Käuferin/Verkäuferin?) Hermannus (Verwalter des Spitals in Brixen, Propst von Neustift 1200–10) 293 (möglicher Verfasser) Iacob (Vetter des Pranttner Michel) Nr. 940, HDS Iohannes, scholasticus aus Isny (de Ysnina) 53, HDS (Adressat einer Urkunde des Bischofs von Brixen) Konrad II. von Rodank (Propst von Neustift ca. 1180–1200) 293 (möglicher Verfasser) Lang, Adamus (Propst von Neustift 1581–85) 726, 76r Mayr (Meyer, Mayer), Caspar 146; 196 (Bibliothekar) Mérey, Sándor 146; 196 Michel auf Ellen 940, HDS Neuhauser, Kaspar (Stadtrichter [1494–1501] und bischöflicher Zöllner [1490/92] in Klausen) 353; 654, 202r (Auftraggeber, Besitzer, Geschenkgeber) Neuhauser, Nikolaus (Kirchpropst [1483/84], Bürgermeister [1488] und Stadtrichter [1492] in Klausen) 353, IIIra Nicolaus Leopoldus, magister ex Insprugga (1511 Lehrer an der Pfarrschule von St. Jakob, Innsbruck, empfängt 1513 in Brixen die niederen Weihen und erhält 1515 Kanonikat, gest. 1535) 167, 1r; 579, 1r (Besitzer) Nikolaus (Propst des Augustiner Chorherrenstiftes St. Dorothea bei Wien) 76 (Schreiber, aber auch Überlasser der Hs.) Posch, Augustin I. (Propst von Neustift, 1519–27) 100 (Auftraggeber, Besitzer) Pranttner, Michel 940, HDS Reger, Johann 654, A (Buchdrucker in Ulm; Druck möglicherweise Vorlage für Miniaturen) Rugen(n), Jörg Neustifter Wappenbuch (mögliche Vorlage) Sacchi, Bartolomeo (Bartolomeo Platina, Bartolomeo Cremonensis) 173 (Notiz am Ende) Schabl, Augustin II. (Propst von Neustift 1569–81) 353; 654 (Besitzer); 726, 76r Scheiber, Nikolaus III. (Propst von Neustift 1439–49) Graduale Neocellense I und II (Auftraggeber) Schobser, Johann 654, A (Buchdrucker in Ulm; Druck möglicherweise Vorlage für Miniaturen) Schuechl, Narziß 353; 405; 654; 15063, E (Buchbinder) Schweidnitz, Nicolaus 614, 150r Servacius Lichtensteyn, de Prussia Graduale Neocellense II, 210v (Buchmaler) Sprenz, Sebastian (Fürstbischof von Brixen 1521–25) 694 (Widmungsträger) Steigenberger, Anton (Propst von Neustift 1737–67) 146 Székely, Samuel 146; 196 (Geschenknehmer) Troyer, Fortunat (Propst von Neustift 1678–1707) 726, 74r Zeiller, Georg (Gregorius) 146 (Dechant – in der Hs. Gregorius, im Chorherrenverzeichnis: Georg) 471 SCHREIBER, SCHREIBERVERSE UND -SPRÜCHE Schreiber Chirchmair, Hainricus (de Malhenstorff, möglicherweise identisch mit Kloster Mallersdorf OSB, Diözese Regensburg) 447, 206vb Claghewinter, Rudolph (de Saxonia) 4, 431vb Edlinger, Heinricus 139 (ohne Nennung des Schreibers); 940, 1r Frechen, Nicolaus 167, 71va Fresell, Iacobus (de Scocia) 617, 53r H., F. 14820, 85v Hölzl, Georg (aus Mattighofen) 353 (ohne Nennung des Schreibernamens); 654, 202r Lämpl, Erasmus 617, 134r, 191v Petrus, Nicolaus 145, 313vb Prensteiner, Franciscus 726, 70v R., H. 142, 25vb, 37va Smyt, Laurencius 550, 445ra Stetner, Stephan 100, 38rb, 93rb, 94va, 104ra; 194, 70v Tegedener, Matheus (de Iuterbock) 155, 105r Weinkern, Albertus 614, 261r Zollner, Friedrich Graduale Neocellense I, 325r; Graduale Neocellense II (ohne Nennung des Schreibernamens) Schreiberverse und -sprüche Assit ad inceptum sancta Maria meum 165, 114r; 168, 1r Detur pro penna scriptori pulchra puella (Colophons Nr. 20587. Walther, Proverbia Nr. 5527a) 605, 5rb Finis adest operis mercedem posco laboris etc. (Colophons Nr. 21270) 52, 4ra, 38va Finito libro grates Christo referamus 327, 19v Finito libro sit laus et gloria Christo. Pitet got umb den der daz geschriben hat 532, 122v Lauda scriptorem donec videas meliorem (Colophons Nr. 22151. Vgl. Walther, Proverbia Nr. 13516a) 605, 5rb Lex est defuncta ubi iudicis est manus uncta 614, 149v 472 ENTSTEHUNGSZEITEN (DATIERUNGEN) 10. Jh. 495 12. Jh. 154 (mit Nachträgen 13. Jh.); 282 (2. Hälfte 12. Jh.); 315; 360; 483 (2. Hälfte) 12./13. Jh. 20; 293; 485 (um 1200) 13. Jh. 19; 86 (wohl Mitte); 107 (1210/34); 327 13./14. Jh. 156 (um 1300); 171; 605; 617, VDS Fragm. 14. Jh. 7 (1. Hälfte); 31; 52, Fälze (Anfang); 52, T. I (Ende); 52, T. II (1396: 270vb), 53 (HDS Fragm.: 2. Hälfte); 99 (1. Hälfte); 161 (Ende); 162 (Ende); 163, T. I (um 1389); 163, T. II (Ende) 168 (9. Jz. 14. Jh.); 253 (1. Hälfte); 293, Makulatureinband; 308; 440, urspr. VDS, heute abgelöst; 532; 606; 617, T. I (7./8. Jz.) 14./15. Jh. 1400 83 (1400: 52rb); 158 (um 1400); 182 (1400: 16rb); 183 (um 1400) 1. H. 15. Jh. 4 (Anfang); 38; 78 (um 1430/33); 106; 145 (1427: 313vb); 146 (Terminus ante quem 1445); 153 (4. Jz.); 163, T. I (um 1443); 165 (um 1407); 165, T. III (1407: 128v); 166 (1. Jz.); 167 (1419: 71va); 173 (2. Drittel); 177 (um 1430/33); 181 (um 1410); 200 (um 1415/20); 217; 447 (1414: 206vb, IX*ra); 453, T. II–IV (um 1440); 614, T. II (u. a. 1419: 261r); 614, T. III (1. Jz.); 614, T. IV (4. Jz.); 748, T. I–III (um 1440); 748, T. IV (um 1430); Graduale Neocellense I (1442: 325v); Graduale Neocellense II (zw. 1442 und 1446) 2. H. 15. Jh. 53 (1452: 237ra); 76, T. I (um 1450); 76, T. II (6. Jz.); 77 (1465: 106vb, 112vb, 119va); 123 (6. Jz.); 125 (6. Jz. 15. Jh.); 142, T. II (1459: 172v); 155 (7. Jz., u. a. 1463: 105r); 160 (um 1540); 174 (um 1470, u. a. 1472: 151rb); 196 (vor 1482?); 204 (um 1470); 252 (4. Viertel); 353 (um 1496); 397; 440 (um 1450); 550, T. I. (6. Jz.); 453, T. I (1455–1460); 550, T. II (7. Jz., u. a. 1465: 97vb, 445ra); 578 (1480: 10vb); 579 (1491: 14v); 597 (um 1464/65); 617, T. I, II, IV (7. Jz., T. I. u a. 1467: 53r); 617, T. III (6. Jz., u. a. 1455: 134r, 191v); 654 (1496: 202r); 754 (6. Jz.); 755 (6. Jz. und 1481?: 3vb); 756 (1479: 53ra); 780 (Ende); 15063 (mit späteren Zusätzen); 139, T. I (1495/96: 1r bzw. 54v); 139, T. II (Ende); 940 (1490/93: 1r bzw. 2r) 15./16. Jh. 190 16. Jh. 85 (Anfang); 100 (1524/26: 1v, 38rb, 50rb, 50v, 93rb, 94va, 104ra); 142, T. I (1521: 4r, 6v, 22va, 29rb, 43vb); 194 (1507: 70v); 405 (1501: 225r); 691 (Anfang); 694 (1523); 726 (1523: 24v, 40v, 65r, 70r, 70v; mit späteren Zusätzen, u. a. 1687: 74r); 755; 776 (Anfang, u. a. 1502: 12v); 14820 (Anfang, mit späteren Zusätzen, u. a. 1578); Neustifter Wappenbuch (1548/52); Graduale Neocellense I (1512: 324r) 473 ENTSTEHUNGSORTE Basel 77 Bologna 7; 99; 217 Bologna s. auch Oberitalien Brixen 53 HDS Fragm.; 550 VD Abklatsch; 578 Brixen s. auch Raum Brixen Entstehungsort unbestimmt 20; 52 Fälze; 282; 315; 360; 405 VDS Fragm.; 483; 485; 495; 532; 605; 606; 617 VDS Fragm.; 1012 Frankreich Cod. 86 (?); 327 (?) Frankreich s. auch Nordfrankreich Italien 173 Neustift s. auch Raum Brixen, Tirol Neustift 76, T. I; 85 (?); 100; 142; 146, T. I (?); 190 (?); 194; 196; 252 (?); 293; 405 (?); 617; 691 (?); 726; 755 (?); 776 (?); 14820; 15063 (?); Graduale Neocellense I und II Nordfrankreich (Raum Paris) 19; 107; 156 Oberitalien s. auch Bologna Oberitalien 4 (Bologna?); 253 (Bologna?); 780 (?) Paris, Raum Paris s. auch Nordfrankreich Pfalzen s. Brixen, Diözese Raum Brixen 78 (Neustift?); 353; 440 urspr. VDS, heute gelöst (?); 550, T. II (u. a. Villanders); 654 Sterzing (?) Neustifter Wappenbuch Süddeutscher Raum 31 (?); 38 (?); 52 (Tirol?); 83; 106 (?); 123; 125; 145 (Tirol?); 146 (T. I: Neustift?); 158; 160; 161; 162; 165; 166; 167; 171; 182; 183; 397 (?); 447; 756 Süddeutsch-österreichischer Raum 308 Schwäbisch-Schweizer Raum 748, T. IV Tirol 4; 174 (?); 177 (Neustift?); 204 (?); 440 (?); 579 (?); 614; 617 (u. a. Neustift); 694; 754 Umfeld Neustift 154; 155 (u. a. Pfalzen); 139; 940 Wien 76, T. II (St. Dorothea); 163; 168 Wiener Raum 53 (?); 153; 181; 200 (?); 453, T. I, T. II–IV (?); 550, T. I; 579 (?); 597; 748, T. I–III (?); 755 (?) ÖSTERREICHISCHE AKADEMIE DER WISSENSCHAFTEN PHILOSOPHISCH-HISTORISCHE KLASSE DENKSCHRIFTEN, 529. BAND VERÖFFENTLICHUNGEN ZUM SCHRIFT- UND BUCHWESEN DES MITTELALTERS ISBN 978-3-7001-8625-0 9 783700 STAMPFER · SCHRETTER-PICKER · DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT REIHE IV, BAND 9 186250 Made in Europe Dph. 529 URSULA STAMPFER – CLAUDIA SCHRETTER-PICKER DIE MITTELALTERLICHEN HANDSCHRIFTEN IN DER BIBLIOTHEK DES AUGUSTINER CHORHERRENSTIFTES NEUSTIFT UNTER MITARBEIT VON PETRA AUSSERLECHNER, GIULIA GABRIELLI, PATRIK KENNEL, GABRIELA KOMPATSCHER GUFLER, WALTER NEUHAUSER, ANNA PINTER, SUSANNE RISCHPLER, MARIA STIEGLECKER, LAV ŠUBARIĆ
https://openalex.org/W4205863874	http://www.ajnr.org/content/ajnr/42/7/1358.full.pdf	English	null	Celebrating 35 Years of the AJNR	American journal of neuroradiology	2,021	cc-by	36	edition Celebrating 35 Years of the AJNR: July 1986 http://www.ajnr.org/content/42/7/1358.citation https://doi.org/10.3174/ajnr.P6837 doi: 2021, 42 (7) 1358 AJNR Am J Neuroradiol of October 23, 2024. This information is current as 35 YEARS AGO Jul 2021 www.ajnr.org 1358
https://openalex.org/W3109274616	https://hal.archives-ouvertes.fr/hal-03040555/file/s13054-020-03402-7.pdf	English	null	Impact of advance directives on the variability between intensivists in the decisions to forgo life-sustaining treatment	Critical care	2,020	cc-by	10,211	Impact of advance directives on the variability between intensivists in the decisions to forgo life-sustaining treatment Margot Smirdec, Mercé Jourdain, Virginie Guastella, Céline Lambert, Jean-Christophe Richard, Laurent Argaud, Samir Jaber, Kada Klouche, Anne Medard, Jean Reignier, et al. To cite this version: Margot Smirdec, Mercé Jourdain, Virginie Guastella, Céline Lambert, Jean-Christophe Richard, et al.. Impact of advance directives on the variability between intensivists in the decisions to forgo life-sustaining treatment. Critical Care, 2020, 24 (1), pp.672. 10.1186/s13054-020-03402-7. hal- 03040555 Distributed under a Creative Commons Attribution 4.0 International License RESEARCH Open Access Impact of advance directives on the variability between intensivists in the decisions to forgo life‑sustaining treatment Margot Smirdec1, Mercé Jourdain2, Virginie Guastella3, Céline Lambert4, Jean‑Christophe Richard5, Laurent Argaud6, Samir Jaber7, Kada Klouche8, Anne Medard9, Jean Reignier10, Jean‑Philippe Rigaud11, Jean‑Marc Doise12, Russell Chabanne13, Bertrand Souweine14, Jeremy Bourenne15, Julie Delmas16, Pierre‑Marie Bertrand17, Philippe Verdier18, Jean‑Pierre Quenot19, Cecile Aubron20, Nathanael Eisenmann21, Pierre Asfar22, Alexandre Fratani23, Jean Dellamonica24, Nicolas Terzi25, Jean‑Michel Constantin26, Axelle Van Lander27,28, Renaud Guerin29, Bruno Pereira4 and Alexandre Lautrette21,30,31* Abstract Background: There is wide variability between intensivists in the decisions to forgo life-sustaining treatment (DFLST). Advance directives (ADs) allow patients to communicate their end-of-life wishes to physicians. We assessed whether ADs reduced variability in DFLSTs between intensivists. Background: There is wide variability between intensivists in the decisions to forgo life-sustaining treatment (DFLST). Advance directives (ADs) allow patients to communicate their end-of-life wishes to physicians. We assessed whether ADs reduced variability in DFLSTs between intensivists. Methods: We conducted a multicenter, prospective, simulation study. Eight patients expressed their wishes in ADs after being informed about DFLSTs by an intensivist-investigator. The participating intensivists answered ten ques‑ tions about the DFLSTs of each patient in two scenarios, referring to patients’ characteristics without ADs (round 1) and then with (round 2). DFLST score ranged from 0 (no-DFLST) to 10 (DFLST for all questions). The main outcome was variability in DFLSTs between intensivists, expressed as relative standard deviation (RSD). Results: A total of 19,680 decisions made by 123 intensivists from 27 ICUs were analyzed. The DFLST score was higher with ADs than without (6.02 95% CI [5.85; 6.19] vs 4.92 95% CI [4.75; 5.10], p < 0.001). High inter-intensivist variability did not change with ADs (RSD: 0.56 (round 1) vs 0.46 (round 2), p = 0.84). Inter-intensivist agreement on DFLSTs was weak with ADs (intra-class correlation coefficient: 0.28). No factor associated with DFLSTs was identified. A qualitative analysis of ADs showed focus on end-of-life wills, unwanted things and fear of pain. Conclusions: ADs increased the DFLST rate but did not reduce variability between the intensivists. In the decision- making process using ADs, the intensivist’s decision took priority. Further research is needed to improve the matching of the physicians’ decision with the patient’s wishes. Trial registration ClinicalTrials.gov Identifier: NCT03013530. Registered 6 January 2017; https://clinicaltrials.gov/ct2/ show/NCT03013530. Correspondence: alautrette@chu‑clermontferrand.fr 21 Intensive Care Unit, Centre Jean Perrin, 54 Rue Montalembert, BP69, 63003 Clermont‑Ferrand, Cedex 1, France Full list of author information is available at the end of the article Correspondence: alautrette@chu‑clermontferrand.fr 21 Intensive Care Unit, Centre Jean Perrin, 54 Rue Montalembert, BP69, 63003 Clermont‑Ferrand, Cedex 1, France Full list of author information is available at the end of the article © The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativeco mmons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. HAL Id: hal-03040555 https://hal.science/hal-03040555v1 Submitted on 2 Jun 2021 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. Distributed under a Creative Commons Attribution 4.0 International License Smirdec et al. Crit Care (2020) 24:672 https://doi.org/10.1186/s13054-020-03402-7 Background de Clermont-Ferrand (IRB00008526; No. 2016/CE87). A consent form was collected from all participants (patients and intensivists) after they had been informed orally and received a written information form. The study was reg- istered on the ClinicalTrials.gov website under number: NCT03013530 in January 2017 and complied with the guidelines of Strengthening the Reporting of Observa- tional Studies in Epidemiology (STROBE) [18]. A decision to forgo life-sustaining treatment (DFLST) is made by 3–30% of intensive care unit (ICU) patients and is recorded in 90% of decedent patients [1]. The DFLST includes decisions about no-escalation or withholding or withdrawal of treatment that lead to differences in mor- tality [2]. These decisions are made by the patient, the physician or close relatives, or result from a shared deci- sion-making process. However, there are numerous limi- tations to this process. A DFLST made by the family can be influenced by their preferences [3] or by the psycho- logical symptoms associated with ICU admission such as anxiety/depression and post-traumatic stress disorder, which prevent the patient’s wishes being clearly reported [4]. In addition, misperceptions about the patient’s prog- nosis by the surrogate can lead to differing expectations by physicians and family and delay decision-making [5, 6]. When making a DFLST, physicians are greatly influ- enced by their personal characteristics including religion and culture [7, 8], which results in considerable variabil- ity in their decisions [7, 9]. This variability is constant within the same specialty or structure [10–14]. Patients want physicians to follow their wishes [15], but most ICU patients are not able to properly communicate these wishes because they lack decision-making capac- ity. Advance directives (ADs) give incapacitated patients the opportunity to indicate what treatment they wish to have [16]. There is a worldwide consensus that physicians should respect the patient’s ADs [17]. In a given setting, when the respect of the patient’s wishes has priority over the personal opinion of the physician, ADs could lead to a decrease in variability in DFLSTs among physicians. However, there is no discussion between the patient and the physician to explain the wishes expressed in ADs. As a result, the physician may interpret the patient’s wishes differently from what was intended. Our study assessed whether ADs, drawn up by patient after receiving infor- mation about DFLSTs and viewing a related video with an intensivist-investigator, would reduce variability in DFLST between intensivists compared to decision-mak- ing without ADs. © The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativeco mmons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Smirdec et al. Crit Care (2020) 24:672 Page 2 of 11 Keywords: Advance directives, Decisions to forgo life-sustaining treatment, ICU Patients and advance directivesh The patients participating in the study were selected from the cohort of consecutive outpatients seen during January–March 2017 for follow-up of chronic disease in the cardiology, pulmonology or nephrology depart- ments of the University Hospital of Clermont-Ferrand. The patient selection criteria were severe stage of chronic cardiac or kidney or respiratory failure associated with comorbid conditions, and life expectancy of less than 5 years according to McCabe score [19], but without acute episode or cognitive impairment on the basis of criteria described by Appelbaum [20]. Of the 23 patients selected, 1 died before the meeting with the intensivist- investigator, 1 was transferred to the palliative care unit, 13 declined and 8 accepted to participate in the study. The characteristics of the 8 patients are shown in Addi- tional file 1. One intensivist-investigator (MS) met each of the eight patients at their home or at a hospital office to provide personalized, clear and full information about DFLSTs and ADs using a video and to suggest that they draw up ADs. The intensivist-investigator had 5 years of ICU practice experience and a master’s diploma in eth- ics. She was trained in communication skills and in face- to-face conversation about ADs. The video, which lasted 10 min, explained ICU life-sustaining treatment, DFLSTs, and the objectives of ADs as laid out in the guidelines of the French Health Authority. During the meeting, the AD forms of the French Health Authority were given to the patient, who had the opportunity to ask questions about DFLSTs, ADs or end-of-life. The patients were then asked questions to check they had understood the information given. The AD forms comprise 11 pages in which patients can designate a surrogate, express their wishes about life- sustaining treatments in the free-text boxes and 15 pages of guidelines about the writing of wishes, and the use of ADs by physicians in accordance with the French law of 2016 [21], which requires physicians to comply with the patients’ wishes except if the ADs are obviously inappro- priate [22]. At the end of the meeting, the patients were invited to draw up their ADs or to take time to discuss Background The other aim of the study was to iden- tify the factors associated with DFLSTs or with change in DFLSTs when ADs were available. Study procedure l l Two clinical scenarios were created by a multidisciplinary team made up of two physicians in palliative care and three intensivists who did not participate in the study. Scenario 1 was followed by six questions and Scenario 2 by four, and both investigated the use of life-sustaining treatment (ICU admission, intubation, renal replace- ment therapy, vasoactive drugs, tracheotomy) for com- munity-acquired pneumonia with septic shock and for septic shock after gastrointestinal surgery, respectively (see Additional file 2). The possible replies were “yes” or “no” for six questions and “yes” or “partially” or “no” for four questions. The replies were rated 1, 0.5 or 0 if they corresponded to a DFLST, a partial DFLST or no DFLST, respectively. The sum of these ten replies made up a DFLST score ranging from 0 if there was no DFLST to 10 if there was DFLST for all questions. The two scenar- ios were submitted online to intensivists from 27 French ICUs in 14 university hospitals and in 9 general hospitals. Each intensivist independently and anonymously com- pleted the questionnaires of the two scenarios for each of eight patients with only the patient’s characteristics avail- able (round 1) and then with the same characteristics and the patient’s ADs available (round 2). Round 2 was sub- mitted to intensivists 2 weeks after completion of round 1 (see Additional file 3). All statistical analyses were performed with Stata soft- ware (version 15, StataCorp, College Station USA, TX). Continuous data were expressed, according to statistical distribution, as mean and standard-deviation or median and interquartile range. The assumption of normality was studied using the Shapiro–Wilk test. Changes in DFLSTs were compared between rounds 1 and 2 with random- effects models taking into account between and within intensivist variability (intensivists as random-effect). Pitman’s test was used to compare RSD. The agreement on DFLSTs between intensivists (inter-agreement) and the within intensivists agreement (intra-agreement) on DFLSTs between the two rounds for each patient were studied using intra-class correlation coefficient (ICC) estimated by the mixed models mentioned above. ICC was interpreted according to standard recommenda- tions: < 0.2 (negligible agreement), 0.20–0.39 (weak agreement), 0.40–0.59 (moderate agreement), 0.60–0.79 (good agreement) and ≥ 0.8 (excellent agreement). Methods We conducted a multicenter, prospective, simulation study, in France from September 2017 to March 2018. The study was approved by the local French ethics com- mittee (Comité de Protection des Personnes Sud-Est VI Smirdec et al. Crit Care (2020) 24:672 Page 3 of 11 DFLST protocol available in ICU and number of DFLSTs per week. with relatives. One patient made a second meeting with the intensivist-investigator to ask further questions. The median time of the meetings was 85.6 [60; 120] min. The ADs were returned within 1 month of the meeting. The number of patients participating in the study was fixed according to the feasibility of answering 20 ques- tions per patient for each round and the case-mix. The sample size of intensivists was fixed to assess how ADs affected the change in variability between intensivists in the DFLSTs. We calculated that at least 120 intensivists evaluating eight patients were necessary to show a 20% relative difference of variability (sample size for a two- sample standard deviations F test), for a two-sided type I error at 5%, a statistical power of 90% and an intra-class correlation coefficient at 0.5, i.e., 960 DFLST scores per round. Study procedure l l To determine the characteristics of the ICU physicians associated with DFLSTs in round 1 or with the change in DFLSTs when ADs were provided, random-effects mod- els (i.e., linear mixed models) were carried out with the following parameters as fixed covariates: age, sex, sta- tus of intensivist, type of hospital, type of ICU, length of practice and religion. The session effect associated with the two measurement times was studied in the same way. The results are expressed in terms of standardized mean differences and 95% confidence intervals. All analyses were performed with a two-sided type I error of 5%. No missing data were observed. Statistical analysish The primary endpoint was variability in DFLSTs between intensivists when ADs were available (round 2) and when they were not (round 1) expressed as the relative stand- ard deviation (RSD; range 0–1; a high rate indicates high inter-intensivist variability). The second endpoint was the identification of factors associated with the DFLSTs or with changes in DFLSTs when ADs were provided. The patient characteristics given to intensivists for the two rounds were age, sex, housing, relatives, medical history, comorbid conditions defined as all diseases or trauma current or previous with the stage or complica- tions reported by the patient or retrieved from medical chart, treatments and dependence in activities of daily living [23]. The recorded characteristics of the intensiv- ists were age, sex, status of intensivist, length of practi- cal experience, speciality of anesthesiology and critical care, religion, interest in ethics, traumatic experience of an end-of-life situation, type of hospital, type of ICU, number of beds and number of intensivists in the ICU, To evaluate the representativeness of the participating intensivists, a sensitivity analysis was performed to com- pare their characteristics with those of the non-partici- pating intensivists. Non-participating intensivists were intensivists contacted but who did not reply to all the questions of the two rounds. A qualitative analysis of ADs was made by a multi- disciplinary team composed of two intensivists, one Smirdec et al. Crit Care (2020) 24:672 Smirdec et al. Crit Care (2020) 24:672 Page 4 of 11 Page 4 of 11 psychologist, one physician in palliative care and two biostatisticians using Alceste software (IMAGE, CNRS, France). The DFLST score was higher with ADs (round 2) than without ADs (round 1) (6.02 95%CI [5.85; 6.19] vs 4.92 95%CI [4.75; 5.10], p < 0.001) (Table 2). There was no sig- nificant change in inter-intensivists variability when ADs were provided (RSD: 0.56 (round 1) vs 0.46 (round 2), p = 0.84) (Table 2). In three patients, the RSD decreased but remained high (Table 2). Inter-intensivists agree- ment on DFLSTs was moderate (ICC = 0.42) without ADs (round 1) and weak (ICC = 0.28) with ADs (round 2). Intra-intensivists agreements on DFLSTs between the two rounds ranged from weak (ICC = 0.22) to moderate (ICC = 0.56) (Table 2). Results Of the 170 intensivists contacted, 123 from 27 ICUs completed the two rounds and made up the study pop- ulation. Their demographic characteristics are given in Table 1. There was no difference between the study population and non-participating intensivists. A total of 19,680 intensivists’ decisions from the two rounds were analyzed. Impact of ADs on the DFLSTs and on the variability in DFLSTsh The proportions of DFLSTs for rounds 1 and 2 are shown in Fig. 1a, b. In the univariate analyses of the overall data of the eight patients, no factor associated with the DFLST score in Table 1 Characteristics of intensivists DFLST decision to forgo life-sustaining treatment, EOL end-of life, ICU intensive care unit, IQR interquartile range Variable Intensivists contacted n = 170 Participating intensivists n = 123 Non-participating intensivists n = 47 p value Male gender, n (%) 113 (63.8) 84 (68.3) 26 (55.3) 0.11 Age (years), mean ± sd 39.9 ± 8.5 40.0 ± 8.5 39.8 ± 8.6 0.88 Status of intensivist, n (%) Assistant 37 (21.8) 24 (19.5) 13 (27.7) Senior 105 (61.8) 78 (63.4) 27 (57.5) 0.51 Professor 28 (16.4) 21 (17.1) 7 (14.8) Length of overall professional experience (years), median [IQR] 6 [3; 15] 7 [3; 15] 6 [3; 15] 0.61 < 5 years, n (%) 60 (35.3) 43 (35.0) 17 (36.2) 5–15 years, n (%) 73 (42.9) 50 (40.6) 23 (48.9) 0.37 > 15 years, n (%) 37 (21.8) 30 (24.4) 7 (14.9) Specialty of anesthesiology and critical care 101 (59.4) 73 (59.3) 28 (59.6) 0.98 Religion, n (%) Catholic 66 (38.8) 47 (38.2) 19 (40.4) Protestant 5 (2.9) 4 (3.3) 1 (2.1) Islam 5 (2.9) 3 (2.4) 2 (4.3) 0.74 Other 10 (5.9) 9 (7.3) 1 (2.1) None 84 (49.5) 60 (48.8) 24 (51.1) Intensivists with an interest in ethics, n (%) 127 (74.7) 90 (73.2) 37 (78.7) 0.56 Intensivists with a traumatic experience of an EOL situation, n (%) 79 (46.5) 57 (46.3) 22 (46.8) 0.96 ICU in university hospital, n (%) 128 (75.3) 94 (76.4) 34 (72.3) 0.58 Type of ICU, n (%) Medical 72 (43.1) 52 (43.0) 20 (43.5) Surgery 20 (12.0) 14 (11.6) 6 (13.0) 0.96 Mixed 75 (44.9) 55 (44.4) 20 (43.5) Number of beds in ICU, median [IQR] 15 [10;18] 15 [10;18] 13 [10; 18] 0.19 Number of intensivists in ICU, median [IQR] 7 [5;10] 7 [5;10] 7 [5;8] 0.09 DFLST Protocol available in ICU, n (%) 112 (65.9) 77 (62.6) 35 (74.5) 0.14 Number of DFLST performed in ICU, n (%) < 1/week 75/158 (47.5) 53/119 (44.5) 22/39 (56.4) 0.20 Table 1 Characteristics of intensivists never stated. The patients expressed themselves in articu- late well-structured sentences. round 1 or with the change in the DFLST score when ADs were available was identified (Table 3). Table 1 Characteristics of intensivists Smirdec et al. Crit Care (2020) 24:672 Page 5 of 11 Fig. 1 Proportions of DFLSTs in round 1 (a) and round 2 (b). Legend: Black bars correspond to DFLSTs; Gray bars correspond to partial DFLSTs; White bars correspond to no-DFLSTs. S1: Scenario 1; S2: scenario 2; Q: Question Fig. 1 Proportions of DFLSTs in round 1 (a) and round 2 (b). Legend: Black bars correspond to DFLSTs; Gray bars correspond to partial DFLSTs; White bars correspond to no-DFLSTs. S1: Scenario 1; S2: scenario 2; Q: Question Table 2 Variability in the DFLST scores (RSD) and the within intensivist agreements (ICC) CI confidence interval, DFLST decision to forgo life-sustaining treatment, RSD relative standard deviation, ICC: intra-class correlation coefficient DFLST score ICC for intensivists Median [95% CI] RSD Round 1 Round 2 p value Round 1 Round 2 p value Patient #1 5.63 [5.20; 6.06] 7.04 [6.62; 7.47] < 0.001 0.43 0.34 0.82 0.48 Patient #2 4.18 [3.84; 4.52] 6.31 [5.86; 6.75] < 0.001 0.46 0.4 0.003 0.22 Patient #3 4.69 [4.29; 5.08] 5.97 [5.53; 6.42] < 0.001 0.47 0.42 0.18 0.47 Patient #4 2.86 [2.54; 3.18] 3.84 [3.45; 4.22] < 0.001 0.61 0.56 0.035 0.25 Patient #5 4.70 [4.25; 5.14] 5.81 [5.43; 6.19] < 0.001 0.53 0.36 0.08 0.38 Patient #6 8.37 [8.02; 8.71] 8.16 [7.76; 8.55] 0.237 0.23 0.27 0.13 0.56 Patient #7 2.42 [2.12; 2.73] 3.72 [3.30; 4.14] < 0.001 0.71 0.63 < 0.001 0.24 Patient #8 6.54 [6.14; 6.95] 7.32 [6.88; 7.76] 0.001 0.35 0.34 0.44 0.37 All patients 4.92 [4.75; 5.10] 6.02 [5.85; 6.19] < 0.001 0.56 0.46 0.84 0.56 ble 2 Variability in the DFLST scores (RSD) and the within intensivist agreements (ICC) Table 2 Variability in the DFLST scores (RSD) and the within intensivist agreements (ICC) never stated. The patients expressed themselves in articu- late well-structured sentences. Discussion Our results show that when provided with ADs, intensiv- ists made more DFLSTs but this did not alter the great variability between them in their decisions. No factor associated with this variability was identified, suggesting multiple causes that were non-specific to the characteris- tics of the intensivists. In addition, the qualitative analysis of ADs highlighted the concerns expressed by the patient A qualitative analysis of ADs was performed on the ver- batim texts amounting to 4091 words. Three themes, accounting for 59% of the words, emerged from the ADs: (1) wills of end-of-life and relatives, (2) unwanted things including therapeutic obstinacy and (3) fear of pain and loss of autonomy (Fig. 2). The most frequently occur- ring words were “life,” “I” and “my.” The word “death” was Smirdec et al. DFLST decision to forgo life-sustaining treatment, EOL end-of life, ICU intensive care unit, SD standard deviation Occurrence of word (number) I My Me Wish Trust Wife Child To see To love Health Priest Relatives To resuscitate 48 37 13 8 5 4 4 3 3 3 3 3 3 Don’t want To prolong To sustain Treatment Don’t wish Person Year Goal Effect Dialysis To refuse Therapeutic obstinacy Support 8 7 6 4 4 4 3 3 3 3 3 3 3 To be Life To dread Condition Suffering Alone Burdensome Loss Burden To put To stay Definitive Vegetative 18 18 6 7 5 4 3 3 3 3 3 3 3 Occurrence of word (number) Fig. 2 Main themes emerging from the advance directives previous reports by the number of questions and the size of the study population, which ensure the robustness of the findings. after they had received full and clear information about DFLST from an intensivist. These results are noteworthy because they show the limitations of the use of ADs.h i g A DFLST is a complex but singular process in which the context, the chances of success, the discomfort of treatment, uncertainty regarding prognosis, potential dis- ability and the wishes of the patient are important con- siderations [30]. ADs with free-text boxes do not allow an exhaustive approach that encompasses in all situations, but give patients the opportunity to express their convic- tions about the physical or mental impairment [31] that could guide intensivists in their choice of DFLSTs. How- ever, the wording used by the patient can be ambiguous or inappropriate to initiate or to withdraw treatment. The applicability of wishes expressed in ADs results from a match between a hypothetical situation and the medi- cal situation affecting the patient [32]. In our study, the ADs rarely indicated specific wishes about life-sustaining treatments despite information about ICU treatments. This finding could explain the lack of impact of ADs on the high variability between intensivists in the DFLSTs. In this qualitative approach, the influence of relatives or physicians was not assessed. Study of these other areas could improve understanding of the drawing-up, accept- ability and usefulness of ADs. The two forms of ADs are complementary and can be associated. After receiv- ing full information during discussion with a healthcare agent, the patient could complete tick-boxes indicating clear DFLSTs that can be implemented whatever the This study highlights the issue of variability between intensivists in the taking of DFLSTs even when ADs are provided. Discussion Crit Care (2020) 24:672 Page 6 of 11 Table 3 Univariate analysis for the identification of factors associated with the DFLSTs and with the change in DFLSTs when advance directives were available DFLST decision to forgo life-sustaining treatment, EOL end-of life, ICU intensive care unit, SD standard deviation Variable DFLST score in round 1 Mean ± SD Correlation coefficient p value Change in DFLST score between round 1 and round 2 Mean ± SD Correlation coefficient p value Gender Female 5.06 ± 2.72 0.42 1.09 ± 2.48 0.96 Male 4.86 ± 2.80 1.10 ± 2.43 Age (years) 0.035 0.46 0.027 0.57 Status of intensivist Assistant 4.91 ± 2.91 0.74 0.98 ± 2.48 0.79 Senior 4.98 ± 2.74 1.15 ± 2.42 Professor 4.74 ± 2.73 1.06 ± 2.53 Length of overall professional experience < 5 years 4.86 ± 2.81 0.23 1.18 ± 2.45 0.42 5–15 years 4.77 ± 2.72 0.94 ± 2.33 > 15 years 5.27 ± 2.79 1.23 ± 2.63 Specialty of anesthesiology and critical care Yes 4.98 ± 2.76 0.62 1.09 ± 2.46 0.75 No 4.86 ± 2.81 1.15 ± 2.42 Religion Catholic 4.94 ± 2.74 0.74 1.18 ± 2.37 0.78 Protestant 4.75 ± 2.61 0.78 ± 1.90 Islam 4.06 ± 2.58 0.73 ± 2.97 Other 4.68 ± 3.06 0.79 ± 2.75 None 5.00 ± 2.77 1.12 ± 2.46 Intensivists with an interest in ethics Yes 5.01 ± 2.80 0.22 1.14 ± 2.49 0.52 No 4.69 ± 2.70 0.99 ± 2.34 Intensivists with a traumatic experience of an EOL situation Yes 4.79 ± 2.73 0.27 1.27 ± 2.44 0.09 No 5.04 ± 2.80 0.95 ± 2.45 Hospital type General 4.74 ± 2.93 0.37 1.33 ± 2.43 0.18 University 4.98 ± 2.72 1.03 ± 2.45 Type of ICU Medical 4.99 ± 2.81 1.12 ± 2.37 Surgery 4.81 ± 2.82 0.89 0.56 ± 2.67 0.14 Mixed 4.90 ± 2.73 1.19 ± 2.45 Number of beds in ICU 0.062 0.61 − 0.10 0.12 Number of intensivists in ICU 0.018 0.87 − 0.05 0.28 DFLST Protocol available in ICU Yes 5.05 ± 2.80 0.16 1.10 ± 2.37 0.92 No 4.71 ± 2.72 1.11 ± 2.57 Number of DFLST performed in ICU < 1/week 4.70 ± 2.84 0.12 1.15 ± 2.62 0.75 ≥ 1/week 5.07 ± 2.69 1.08 ± 2.26 Table 3 Univariate analysis for the identification of factors associated with the DFL when advance directives were available DFLST decision to forgo life-sustaining treatment, EOL end-of life, ICU intensive care unit, SD standard deviation Variable DFLST score in round 1 Mean ± SD Correlation coefficient p value Change between Mean ± Correlat Gender Female 5.06 ± 2.72 0.42 1.09 ± 2.4 Male 4.86 ± 2.80 1.10 ± 2.4 Age (years) 0.035 0.46 0.027 Status of intensivist Assistant 4.91 ± 2.91 0.74 0.98 ± 2.4 Senior 4.98 ± 2.74 1.15 ± 2.4 Professor 4.74 ± 2.73 1.06 ± 2.5 Length of overall professional experience < 5 years 4.86 ± 2.81 0.23 1.18 ± 2.4 5–15 years 4.77 ± 2.72 0.94 ± 2.3 > 15 years 5.27 ± 2.79 1.23 ± 2.6 Specialty of anesthesiology and critical care Yes 4.98 ± 2.76 0.62 1.09 ± 2.4 No 4.86 ± 2.81 1.15 ± 2.4 Religion Catholic 4.94 ± 2.74 0.74 1.18 ± 2.3 Protestant 4.75 ± 2.61 0.78 ± 1.9 Islam 4.06 ± 2.58 0.73 ± 2.9 Other 4.68 ± 3.06 0.79 ± 2.7 None 5.00 ± 2.77 1.12 ± 2.4 Intensivists with an interest in ethics Yes 5.01 ± 2.80 0.22 1.14 ± 2.4 No 4.69 ± 2.70 0.99 ± 2.3 Intensivists with a traumatic experience of an EOL situation Yes 4.79 ± 2.73 0.27 1.27 ± 2.4 No 5.04 ± 2.80 0.95 ± 2.4 Hospital type General 4.74 ± 2.93 0.37 1.33 ± 2.4 University 4.98 ± 2.72 1.03 ± 2.4 Type of ICU Medical 4.99 ± 2.81 1.12 ± 2.3 Surgery 4.81 ± 2.82 0.89 0.56 ± 2.6 Mixed 4.90 ± 2.73 1.19 ± 2.4 Number of beds in ICU 0.062 0.61 − 0.10 Number of intensivists in ICU 0.018 0.87 − 0.05 DFLST Protocol available in ICU Yes 5.05 ± 2.80 0.16 1.10 ± 2.3 No 4.71 ± 2.72 1.11 ± 2.5 Number of DFLST performed in ICU < 1/week 4.70 ± 2.84 0.12 1.15 ± 2.6 ≥ 1/week 5.07 ± 2.69 1.08 ± 2.2 Page 7 of 11 Smirdec et al. Discussion Crit Care (2020) 24:672 Unwanted things Fear of pain and loss of autonomy “I want my wife and children to be with me in my final moments”. / “I want a priest to be present at my deathbed”. “I confirm my refusal of all treatment and therapeutic obstinacy whose sole aim is to keep me artificially alive”. “I am afraid of unnecessary suffering if I’m kept alive in a vegetative state”. / “I fear that at the end of my life I will be a burden to society”. I My Me Wish Trust Wife Child To see To love Health Priest Relatives To resuscitate 48 37 13 8 5 4 4 3 3 3 3 3 3 Don’t want To prolong To sustain Treatment Don’t wish Person Year Goal Effect Dialysis To refuse Therapeutic obstinacy Support 8 7 6 4 4 4 3 3 3 3 3 3 3 To be Life To dread Condition Suffering Alone Burdensome Loss Burden To put To stay Definitive Vegetative 18 18 6 7 5 4 3 3 3 3 3 3 3 Wills of end-of-life and relatives Occurrence of word (number) Examples of sentences Themes Fig. 2 Main themes emerging from the advance directives Examples of sentences “I confirm my refusal of all treatment and therapeutic obstinacy whose sole aim is to keep me artificially alive”. “I am afraid of unnecessary suffering if I’m kept alive in a vegetative state”. / “I fear that at the end of my life I will be a burden to society”. “I confirm my refusal of all treatment and therapeutic obstinacy whose sole aim is to keep me artificially alive”. Occurrence of word (number) Great variability has been previously reported in several studies [12, 13, 24–27] and attributed to indi- vidual physician characteristics such as religion, cul- ture and geographic regions [7, 8]. The goal of ADs is to enable physicians to comply with the patient’s wishes or request, irrespective of their own personal character- istics [15, 16]. In theory, therefore, ADs should elimi- nate variability but our findings showed that physicians disagreed about DFLSTs even when ADs were avail- able. There are several possible explanations of this find- ing. First, the intensivists did not use the ADs to make DFLSTs. However, DFLSTs increased when ADs were available. Second, ADs expressed in free-text boxes led to more possible interpretations of patients’ wishes than ADs drawn up in tick-box form, which includes the use of medical terms for instructions that are easily understood by intensivists and provide clear answers to DFLSTs [28]. In studies assessing the interrater reliability for each section of the POLST form using a binary “yes/no” approach, the Κappa coefficients varied from 0.70 to 1.00 [29]. It is possible that the variability among intensivists is low in the binary “yes/no” approach of DFLSTs wishes. However, the possible replies in our study were “yes,” “no” and sometimes “partial.” Our study differs from these Smirdec et al. Crit Care (2020) 24:672 Smirdec et al. Crit Care (2020) 24:672 Page 8 of 11 Page 8 of 11 situation and use free-text boxes to express values that could help guide the physician in making DFLSTs in sit- uations not previously considered. The decision about resuscitation status is easily made by the patient and can be registered in a tick-box. In contrast, the initiation of renal replacement therapy is an abstract concept that is discussed according to clinical condition and for which the expression of value is more relevant. The surrogate can add nuances and clarifications to the values relayed in ADs so that the best decisions are made according to the specific condition and prognosis of the patient [33]. A surrogate involved in the drafting of values could limit the influence of the intensivist in the decision-making process using ADs [34]. Alternatively, the physician’s interpretation of values could be assessed in clinical sce- narios. Occurrence of word (number) Feedback on DFLSTs made by a sample of inten- sivists in clinical scenarios would allow the patient to modify ADs so that the intensivists’ decisions comply with her or his own and thereby reduce inter-intensivist variability. Supplementary Information The online version contains supplementary material available at https://doi. org/10.1186/s13054-020-03402-7. Additional file 1. Characteristics of the 8 patients who wrote advance directives (table). Additional file 2. The two clinical scenarios (text). Additional file 3. Flow chart and timing of the study (Figure). Abbreviations ADs: Advance directives; DFLST: Decision to forgo life-sustaining treatment; CI: Confidence intervals; ICC: Intra-class correlation coefficient; ICU: Intensive care unit; POLST: Physician orders for life-sustaining treatment; RSD: Relative standard deviation. y Our study has a number of limitations. First, our anal- yses were performed on the ADs of eight patients. This sample, in agreement with the study feasibility assess- ment, is not an exhaustive representation of the French population. Nevertheless, the ADs in the study were real documents, which could be used in the decision-making process in clinical practice [35]. Second, to standard- ize the process, only one intensivist-investigator briefed the patients about DFLSTs and ADs. However, this approach entails the risk of personal influence by the clinician, which can lead to cognitive bias. Third, we do not rule out that the DFLSTs made by intensivists in the scenarios could differ from those taken in everyday practice. However, only a simulation study provides the standardization of situations, which allows assessment of variability between physicians. Our simulation study was time-consuming and unpaid, which could have restricted the participation of some intensivists. The intensivists taking part may have had an interest in ethics and hence were perhaps not representative of the profession as a whole. Fourth, our study design gave no information about family and ICU team discussions or staff opinion. Many DFLSTs are made with non-intensivist physicians, relatives or nurses. Their influence on DFLSTs was not assessed in our study, which focused on the intensivists because they are the main decision-makers for DFLST in the ICU. Fifth, most ADs were collected within 1 month of a single meeting which for some patients could have been too short time to formulate their wishes in full. However, the information about ADs, given by an inten- sivist-investigator with a video, was of a quality as high as that provided in clinical practice [36] and similar to that used in reference publications [33, 37]. Conclusions Our study shows that when ADs were available, intensiv- ists were more likely to make DFLSTs. However, ADs did not reduce high inter-intensivist variability in the deci- sion-making process. The great variability observed show that the intensivist’s preferences had priority. Further research is needed to establish a process that achieves a better matching of the physicians’ decisions with the patient’s wishes. Acknowledgements g We are indebted to Mr. Jeffrey Watts for assistance in the preparation of the manuscript. Members of the Facing group: Geoffrey Ledoux (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation Lille); Lea Satrebuisson (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Sophie Six (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Céline Dupre (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Patrick Girardie (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Anahita Rouze (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Erika Decrucq-Parmentier (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Daniel Mathieu (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Julien Poissy (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Sébastien Préau (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Emmanuelle Jaillette (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Raphaël Favory (Centre Hospitalier Universitaire de Lille, Médecine Intensive Réanimation, Lille); Honade Yonis (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Romain Hernu (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Marie Simon (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Thomas Baudry (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Thomas Madelaine (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Adeline Grateau (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Martin Cour (Centre Hospitalier Universitaire de Lyon, Médecine Intensive Réanimation, Lyon); Gérald Chanques (Centre Hospitalier Universi‑ taire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Julie Carr (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Matthieu Conseil (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Audrey De Jong (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Jean-Marc Delay (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Etienne Imhoff (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Bruno Souche (Centre Hospitalier Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Marina Thirion (Centre Hospitalier Page 9 of 11 Smirdec et al. Crit Care (2020) 24:672 Smirdec et al. Crit Care (2020) 24:672 Smirdec et al. Acknowledgements Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Aulère Buzançais (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Boris Jung (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Vincent Bruno (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Delphine Daubin (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Laura Platon (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Jean-Jacques Tudesq (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Liliane Landreau (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Henri Boby (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Nicolas Rascol (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Lucia Uta (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Vedat Eljezi (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Mélanie Chasteloux (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Nicolas Coullier (Centre Hospitalier Universitaire de Clermont-Ferrand, A thé i t Ré i ti Chi i l Cl t F d) L t Ni l t ( p , Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laurent Nicolet (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Charlotte Garret (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Noëlle Brule (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Agathe Delbove (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Maelle Martin (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Jean-Batptiste Lascarrou (Centre Hospitalier Universi‑ taire de Nantes, Médecine Intensive Réanimation, Nantes); François Bougerol (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Pierre-Louis Declercq (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Marion Beuzelin (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Jean-Pierre Eraldi (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Antoine Marchalot (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Thomas Poussant (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Maël Hamet (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur- Saône); Martial Delorme (Centre Hospitalier de Chalon-sur-Saône, Réanima‑ tion, Chalon-sur-Saône); Adrien Lhoumeau (Centre Hospitalier de Chalon-sur- Saône, Réanimation, Chalon-sur-Saône); Jen Darphin (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Julien Pascal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Charlotte Fernandez-Canal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Aurélien Momon (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Julien Marin (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laure Calvet (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanima‑ tion, Clermont-Ferrand); Edouard Soum (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanimation, Clermont-Ferrand); Claire Bachelier (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanimation, Clermont-Ferrand); Kostas Bachoumas (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanima‑ tion, Clermont-Ferrand); Thomas Godet (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Danièle Saurel (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Marie Vignaud (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Marc Gainnier (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Dominique Lambert (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Lucas Benarous (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Julien Carvelli (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Philippe Letocart (Centre Hospitalier de Rodez, Médecine Intensive Réanimation, Rodez); Arnaud Delahaye (Centre Hospitalier de Rodez, Médecine Intensive Réanimation, Rodez); Sophie Dufraisse (Centre Hospitalier de Rodez, Médecine Intensive Réanimation, Rodez); François Lenfant (Centre Hosptialier de Cannes, Médecine Intensive Réanimation, Cannes); Raphaël Chambon (Centre Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laurent Nicolet (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Charlotte Garret (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Noëlle Brule (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Agathe Delbove (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Maelle Martin (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Jean-Batptiste Lascarrou (Centre Hospitalier Universi‑ taire de Nantes, Médecine Intensive Réanimation, Nantes); François Bougerol (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Pierre-Louis Declercq (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Marion Beuzelin (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Jean-Pierre Eraldi (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Antoine Marchalot (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Thomas Poussant (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Maël Hamet (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur- Saône); Martial Delorme (Centre Hospitalier de Chalon-sur-Saône, Réanima‑ tion, Chalon-sur-Saône); Adrien Lhoumeau (Centre Hospitalier de Chalon-sur- Saône, Réanimation, Chalon-sur-Saône); Jen Darphin (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Julien Pascal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Charlotte Fernandez-Canal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Aurélien Momon (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Julien Marin (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laure Calvet (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanima‑ tion Clermont-Ferrand); Edouard Soum (Centre Hospitalier Universitaire de Consent for publication Not applicable. Not applicable. Competing interests The others authors have no financial conflict of interest related to this study. Availability of data and materialsi The data that support the findings of this study are available from the cor‑ responding author upon reasonable request. Authors’ contributions MS, AL and BP conceived and designed the study; MS, MJ, JCR, LA, SJ, KK, AM, JR, JPR, JMD, RC, BS, JB, JD, PMB, PV, JPQ, CA, NE, PA, AF, JD, NT, JMC and RG recruited the participants and collected the data; MS, AVL, VG, CL, RG, AL and BP analyzed and interpreted data; MS, AL and BP drafted the report and all authors contributed to review it. All authors approved the final version. All authors read and approved the final manuscript. Acknowledgements Crit Care (2020) 24:672 Universitaire de Montpellier, Anesthésie et Réanimation Chirurgicale, Montpellier); Aulère Buzançais (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Boris Jung (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Vincent Bruno (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Delphine Daubin (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Laura Platon (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Jean-Jacques Tudesq (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Liliane Landreau (Centre Hospitalier Universitaire de Montpellier, Médecine Intensive Réanimation, Montpellier); Henri Boby (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Nicolas Rascol (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Lucia Uta (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Vedat Eljezi (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Mélanie Chasteloux (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Nicolas Coullier (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laurent Nicolet (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Charlotte Garret (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Noëlle Brule (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Agathe Delbove (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Maelle Martin (Centre Hospitalier Universitaire de Nantes, Médecine Intensive Réanimation, Nantes); Jean-Batptiste Lascarrou (Centre Hospitalier Universi‑ taire de Nantes, Médecine Intensive Réanimation, Nantes); François Bougerol (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Pierre-Louis Declercq (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Marion Beuzelin (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Jean-Pierre Eraldi (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Antoine Marchalot (Centre Hospitalier de Dieppe, Réanimation, Dieppe); Thomas Poussant (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Maël Hamet (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur- Saône); Martial Delorme (Centre Hospitalier de Chalon-sur-Saône, Réanima‑ tion, Chalon-sur-Saône); Adrien Lhoumeau (Centre Hospitalier de Chalon-sur- Saône, Réanimation, Chalon-sur-Saône); Jen Darphin (Centre Hospitalier de Chalon-sur-Saône, Réanimation, Chalon-sur-Saône); Julien Pascal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Charlotte Fernandez-Canal (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Aurélien Momon (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Julien Marin (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Laure Calvet (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanima‑ tion, Clermont-Ferrand); Edouard Soum (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanimation, Clermont-Ferrand); Claire Bachelier (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanimation, Clermont-Ferrand); Kostas Bachoumas (Centre Hospitalier Universitaire de Clermont-Ferrand, Médecine Intensive Réanima‑ tion, Clermont-Ferrand); Thomas Godet (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Danièle Saurel (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Marie Vignaud (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Marc Gainnier (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Dominique Lambert (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Lucas Benarous (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Julien Carvelli (Centre Hospitalier Universitaire de Marseille, Médecine Intensive Réanimation, Marseille); Philippe Letocart (Centre H it li d R d Méd i I t i Ré i ti R d ) A d Hospitalier de Cannes, Médecine Intensive Réanimation, Cannes); Jerôme Bedel (Centre Hospitalier de Cannes, Médecine Intensive Réanimation, Cannes); Nicolas François (Centre Hospitalier de Montluçon, Réanimation, Montluçon); Thierry Comte (Centre Hospitalier de Montluçon, Réanimation, Montluçon); Pierre Couhault (Centre Hospitalier de Montluçon, Réanimation, Montluçon); Jean-Baptiste Roudaut (Centre Hospitalier Universitaire de Dijon, Médecine Intensive Réanimation, Dijon); Auguste Dargent (Centre Hospitalier Universitaire de Dijon, Médecine Intensive Réanimation, Dijon); Gwenaël Prat (Centre Hospitalier Universitaire de Brest, Médecine Intensive Réanimation, Brest); Vanessa Jean-Michel (Centre Hospitalier Universitaire de Brest, Médecine Intensive Réanimation, Brest); Beatrice Fourgeaud (Centre de Lutte Contre le Cancer Jean Perrin, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Charlotte Baud (Centre de Lutte Contre le Cancer Jean Perrin, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Cédric Darreau (Centre Hospitalier Universitaire de Angers, Médecine Intensive Réanimation, Angers); Pierre-Yves Olivier (Centre Hospitalier Universitaire de Angers, Médecine Intensive Réanimation, Angers); Mourad Benyamina (Centre Hosptitalier Universitaire Saint-Louis, Centre de traitement des Brûlés, Paris); François Depret (Centre Hosptitalier Universitaire Saint-Louis, Centre de traitement des Brûlés, Paris); Hervé Hyvernat (Centre Hospitalier Universitaire de Nice, Médecine Intensive Réanimation, Nice); Pierre-Eric Danin (Centre Hospitalier Universitaire de Nice, Médecine Intensive Réanimation, Nice); Nordine Ait Ben Said (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Raiko Blondonnet (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Clémence Deville (Centre Hospitalier Universitaire de Clermont-Ferrand, Nephrology, Clermont-Ferrand); Aurélien Mulliez (Centre Hospitalier Universitaire de Clermont-Ferrand, Service de Biostastistiques, Clermont-Ferrand); Pierre Cornillon (Clermont-Ferrand); David Brugnon (Centre Hospitalier Universitaire de Clermont-Ferrand, Soins palliatifs, Clermont-Ferrand); Lise Vernis (Centre Hospitalier Universitaire de Clermont-Ferrand, Anesthésie et Réanimation Chirurgicale, Clermont-Ferrand); Delphine Aguettaz (Centre Hospitalier Universitaire de Clermont-Ferrand, Cardiologie, Clermont-Ferrand); Lise Thibonnier (Centre Hospitalier Universi‑ taire de Clermont-Ferrand, Pneumologie, Clermont-Ferrand); Hugues François (Centre Hospitalier Universitaire de Clermont-Ferrand, Pneumologie, Clermont-Ferrand); Frédérique Martins-Condé (Centre Hospitalier Universitaire de Clermont-Ferrand, Gériatrie, Clermont-Ferrand); Maeva Wongfat (Pole santé république, Néphrologie, Clermont-Ferrand); Ibrahim Moustapha (Centre Hospitalier Universitaire de Grenoble, Médecine Intensive Réanimation, Grenoble); Thibaud Pranal (Centre Hospitalier de Vichy, Réanimation, Vichy); François Brenat (Centre Hospitalier du Puy-en-Velay, Réanimation, Le Puy-en-Velay); Bertrand Combres (Centre Hospitalier d’Albi, Réanimation, Albi); Anne Veinstein (Centre Hospitalier Universitaire de Poitiers, Médecine Intensive Réanimation, Poitiers); Alexandre Boyer (Centre Hospitalier Universitaire de Bordeaux, Médecine Intensive Réanimation, Bordeaux). Ethical approval and consent to participate Ethical approval and consent to participate Approved by the Institutional Review Board of the Comité Pour la Protection des Personnes Sud-Est 6 (IRB00008526; No.2016/CE87). Approved by the Institutional Review Board of the Comité Pour la Protection des Personnes Sud-Est 6 (IRB00008526; No.2016/CE87). Funding No Grant. No Grant. Author details 1 between physicians and surrogate decision makers of critically ill patients. JAMA. 2016;315:2086–94. between physicians and surrogate decision makers of critically ill patients. JAMA. 2016;315:2086–94. 1 Department of Anaesthesiology and Critical Care Medicine, Estaing Hospital, University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 2 INSERM U1190, CHU Lille, Department of Critical Care Medicine, Roger Salengro Hospi‑ tal, Univ. Lille, 59000 Lille, France. 3 Palliative Care Unit, Louise Michel Hospital, University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 4 Biosta‑ tistics Unit (DRCI), University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 5 Medical Intensive Care Unit, La Croix Rousse Hospital, University Hospital of Lyon, Lyon, France. 6 Medical Intensive Care Unit, Edouard Herriot Hospital, University Hospital of Lyon, Lyon, France. 7 Department of Anaes‑ thesiology and Critical Care Medicine, Saint Eloi Hospital, University Hospital of Montpellier, Montpellier, France. 8 Medical Intensive Care Unit, Lapeyron‑ nie Hospital, University Hospital of Montpellier, Montpellier, France. 9 Cardiac Surgery Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Montpied Hospital, University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 10 Medical Intensive Care Unit, Hotel‑Dieu Hospital, University Hospital of Nantes, Nantes, France. 11 Intensive Care Unit, Pasteur Hospital, Hospital of Dieppe, Dieppe, France. 12 Intensive Care Unit, Morey Hospital, Hospital of Chalon-Sur-Saône, Chalon‑sur‑Saône, France. 13 Neuro‑ critical Care Unit, Department of Anaesthesiology and Critical Care Medicine, Montpied Hospital, University Hospital of Clermont-Ferrand, Clermont‑Fer‑ rand, France. 14 Medical Intensive Care Unit, Montpied Hospital, University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 15 Emergency Intensive Care Unit, La Timone Hospital, University Hospital of Marseille, Mar‑ seille, France. 16 Intensive Care Unit, Puel Hospital, Hospital of Rodez, Rodez, France. 17 Intensive Care Unit, Veil Hospital, Hospital of Cannes, Cannes, France. 18 Intensive Care Unit, Hospital of Montluçon, Montluçon, France. 19 Medical Intensive Care Unit, Mitterrand Hospital, University Hospital of Dijon, Dijon, France. 20 Medical Intensive Care Unit, Centre Hospitalier Universitaire de Brest, Université de Bretagne Occidentale, Brest, France. 21 Intensive Care Unit, Centre Jean Perrin, 54 Rue Montalembert, BP69, 63003 Clermont‑Ferrand, Cedex 1, France. 22 Medical Intensive Care Unit, Larrey Hospital, University Hospital of Angers, Angers, France. 23 Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Saint‑Louis Hospital, Assistance Publique Hopitaux de Paris, Paris, France. 24 Medical Intensive Care Unit,l’Archet Hospital, University Hospital of Nice, Nice, France. 25 Medical Intensive Care Unit, Michallon Hospital, University Hospital of Grenoble, Grenoble, France. 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J Officiel 2016;0181 23. Katz S. Assessing self-maintenance: activities of daily living, mobility, and instrumental activities of daily living. J Am Geriatr Soc. 1983;31:721–7. Author details 1 26 GRC 29, AP‑HP, DMU DREAM, Department of Anaesthesiology and Criti‑ cal Care, Pitié‑Salpêtrière Hospital, Sorbonne University, Paris, France. 27 UPU ACCePPt, Université Clermont Auvergne, Clermont‑Ferrand, France. 28 EA‑481, Laboratoire de Neurosciences, UBFC, Besançon, France. 29 Intensive Care Unit, Department of Anaesthesiology and Critical Care Medicine, Estaing Hospital, University Hospital of Clermont-Ferrand, Clermont‑Ferrand, France. 30 LMGE «Laboratoire Micro‑Organismes: Génome Et Environnement», UMR CNRS 6023, Clermont-Auvergne University, Clermont‑Ferrand, France. 31 Intensive Care Medicine, Montpied Teaching Hospital, 54 Rue Montalembert, BP69, 63003 Clermont‑Ferrand, Cedex 1, France. p 6. Luce JM. A history of resolving conflicts over end-of-life care in intensive care units in the United States. Crit Care Med. 2010;38:1623–9. 7. Sprung CL, Cohen SL, Sjokvist P, Baras M, Bulow H-H, Hovilehto S, et al. End-of-life practices in European intensive care units: the Ethicus Study. JAMA. 2003;290:790–7. p g , , j , , , , End-of-life practices in European intensive care units: the Ethicus Study. JAMA. 2003;290:790–7. 8. Sprung CL, Woodcock T, Sjokvist P, Ricou B, Bulow H-H, Lippert A, et al. Reasons, considerations, difficulties and documentation of end-of-life decisions in European intensive care units: the ETHICUS Study. Intensive Care Med. 2008;34:271–7. 9. Valley TS, Admon AJ, Zahuranec DB, Garland A, Fagerlin A, Iwashyna TJ. Estimating ICU benefit: a randomized study of physicians. Crit Care Med. 2019;47:62–8. 10. Barnato AE, Hsu HE, Bryce CL, Lave JR, Emlet LL, Angus DC, et al. Using simulation to isolate physician variation in intensive care unit admission decision making for critically ill elders with end-stage cancer: a pilot feasibility study. Crit Care Med. 2008;36:3156–63. y y 11. Kruser JM, Aaby DA, Stevenson DG, Pun BT, Balas MC, Barnes-Daly MA, et al. Assessment of variability in end-of-life care delivery in intensive care units in the United States. JAMA Netw Open. 2019;2:e1917344. 12. Hart JL, Harhay MO, Gabler NB, Ratcliffe SJ, Quill CM, Halpern SD. Vari‑ ability among US intensive care units in managing the care of patients admitted with preexisting limits on life-sustaining therapies. JAMA Intern Med. 2015;175:1019–26. 13. Curtis JR, Engelberg RA, Teno JM. Understanding variability of end-of-life care in the ICU for the elderly. Intensive Care Med. 2017;43:94–6. 14. Wunsch H, Harrison DA, Harvey S, Rowan K. End-of-life decisions: a cohort study of the withdrawal of all active treatment in intensive care units in the United Kingdom. Intensive Care Med. 2005;31:823–31. 15. Silveira MJ, Kim SYH, Langa KM. References Crit Care (2020) 24:672 Smirdec et al. Crit Care (2020) 24:672 Page 11 of 11 29. Lovadini GB, Fukushima FB, Schoueri JFL, Dos Reis R, Fonseca CGF, Rodri‑ guez JJC, et al. Evaluation of the interrater reliability of end-of-life medical orders in the physician orders for life-sustaining treatment form. JAMA Netw Open. 2019;2:e192036. expressed treatment preferences in intensive care units. Am J Respir Crit Care Med. 2017;196:242–5. 35. Council of Europe. Recommendation CM/Rec(2009) 11 of the Com‑ mittee of Ministers to member states on principles continuing powers of attorney and advance directives for incapacity. Eur J Health Law. 2010;17:205–10. 30. Guidet B, Hodgson E, Feldman C, Paruk F, Lipman J, Koh Y, et al. The Durban World Congress Ethics Round Table Conference Report: II. With‑ holding or withdrawing of treatment in elderly patients admitted to the intensive care unit. J Crit Care. 2014;29:896–901. 36. Miki R, Becker CB, Ide K, Kawakami K. Timing and facilitation of advanced directives in Japan. Arch Gerontol Geriatr. 2018;79:83–7. 31. Gaster B, Larson EB, Curtis JR. Advance directives for dementia: meeting a unique challenge. JAMA. 2017;318:2175–6. 37. Philippart F, Vesin A, Bruel C, Kpodji A, Durand-Gasselin B, Garçon P, et al. The ETHICA study (part I): elderly’s thoughts about intensive care unit admission for life-sustaining treatments. Intensive Care Med. 2013;39:1565–73. 32. Hartog CS, Peschel I, Schwarzkopf D, Curtis JR, Westermann I, Kabisch B, et al. Are written advance directives helpful to guide end-of-life therapy in the intensive care unit? A retrospective matched-cohort study. J Crit Care. 2014;29:128–33. References 24. Mark NM, Rayner SG, Lee NJ, Curtis JR. Global variability in withholding and withdrawal of life-sustaining treatment in the intensive care unit: a systematic review. Intensive Care Med. 2015;41:1572–85. 1. Lobo SM, De Simoni FHB, Jakob SM, Estella A, Vadi S, Bluethgen A, et al. Decision-making on withholding or withdrawing life support in the ICU: a worldwide perspective. Chest. 2017;152:321–9. 1. Lobo SM, De Simoni FHB, Jakob SM, Estella A, Vadi S, Bluethgen A, et al. Decision-making on withholding or withdrawing life support in the ICU: a worldwide perspective. Chest. 2017;152:321–9. 25. Alemayehu E, Molloy DW, Guyatt GH, Singer J, Penington G, Basile J, et al. Variability in physicians’ decisions on caring for chronically ill elderly patients: an international study. CMAJ Can Med Assoc J J Assoc Medicale Can. 1991;144:1133–8. 2. Lautrette A, Garrouste-Orgeas M, Bertrand P-M, Goldgran-Toledano D, Jamali S, Laurent V, et al. Respective impact of no escalation of treatment, withholding and withdrawal of life-sustaining treatment on ICU patients’ prognosis: a multicenter study of the Outcomerea Research Group. Intensive Care Med. 2015;41:1763–72. 2. Lautrette A, Garrouste-Orgeas M, Bertrand P-M, Goldgran-Toledano D, Jamali S, Laurent V, et al. Respective impact of no escalation of treatment, withholding and withdrawal of life-sustaining treatment on ICU patients’ prognosis: a multicenter study of the Outcomerea Research Group. Intensive Care Med. 2015;41:1763–72. 26. Wunsch H, Angus DC, Harrison DA, Collange O, Fowler R, Hoste EAJ, et al. Variation in critical care services across North America and Western Europe. Crit Care Med. 2008;36(2787–93):e1-9. 3. Ciroldi M, Cariou A, Adrie C, Annane D, Castelain V, Cohen Y, et al. Ability of family members to predict patient’s consent to critical care research. Intensive Care Med. 2007;33:807–13. 27. Quill CM, Ratcliffe SJ, Harhay MO, Halpern SD. Variation in decisions to forgo life-sustaining therapies in US ICUs. Chest. 2014;146:573–82. 4. Lautrette A, Peigne V, Watts J, Souweine B, Azoulay E. Surrogate decision makers for incompetent ICU patients: a European perspective. Curr Opin Crit Care. 2008;14:714–9. 28. Halpern SD, Small DS, Troxel AB, Cooney E, Bayes B, Chowdhury M, et al. Effect of default options in advance directives on hospital-free days and care choices among seriously ill patients: a randomized clinical trial. JAMA Netw Open. 2020;3:e201742. 5. White DB, Ernecoff N, Buddadhumaruk P, Hong S, Weissfeld L, Curtis JR, et al. Prevalence of and factors related to discordance about prognosis Smirdec et al. Publisher’s Note 33. Garrouste-Orgeas M, Tabah A, Vesin A, Philippart F, Kpodji A, Bruel C, et al. The ETHICA study (part II): simulation study of determinants and variabil‑ ity of ICU physician decisions in patients aged 80 or over. Intensive Care Med. 2013;39:1574–83. Springer Nature remains neutral with regard to jurisdictional claims in pub‑ lished maps and institutional affiliations. 34. Chiarchiaro J, Ernecoff NC, Scheunemann LP, Hough CL, Carson SS, Peterson MW, et al. Physicians rarely elicit critically ill patients’ previously 34. Chiarchiaro J, Ernecoff NC, Scheunemann LP, Hough CL, Carson SS, Peterson MW, et al. Physicians rarely elicit critically ill patients’ previously • fast, convenient online submission • thorough peer review by experienced researchers in your ﬁeld • rapid publication on acceptance • support for research data, including large and complex data types • gold Open Access which fosters wider collaboration and increased citations maximum visibility for your research: over 100M website views per year • At BMC, research is always in progress. Learn more biomedcentral.com/submissions Ready to submit your research Ready to submit your research ? Choose BMC and benefit from: ? 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https://openalex.org/W2760425327	http://shura.shu.ac.uk/17003/1/Mountanin%20Long-term%2024-month%20effect%20on%20health%20and%20well-being%20of%20the%20Lifestyle%20Matters.pdf	English	null	The long-term (24-month) effect on health and well-being of the Lifestyle Matters community-based intervention in people aged 65 years and over: a qualitative study	BMJ open	2,017	cc-by	9,005	The long-term (24-month) effect on health and well-being of the Lifestyle Matters community-based intervention in people aged 65 years and over: a qualitative study CHATTERS, Robin, ROBERTS, Jennifer, MOUNTAIN, Gail, COOK, Sarah, WINDLE, Gill, CRAIG, Claire <http://orcid.org/0000-0002-3475-3292> and PRANGE, Kirsty CHATTERS, Robin, ROBERTS, Jennifer, MOUNTAIN, Gail, COOK, Sarah, WINDLE, Gill, CRAIG, Claire <http://orcid.org/0000-0002-3475-3292> and SPRANGE, Kirsty Available from Sheffield Hallam University Research Archive (SHURA) at: http://shura.shu.ac.uk/17003/ Available from Sheffield Hallam University Research Archive (SHURA) at: http://shura.shu.ac.uk/17003/ This document is the author deposited version. You are advised to consult the publisher's version if you wish to cite from it. Published version CHATTERS, Robin, ROBERTS, Jennifer, MOUNTAIN, Gail, COOK, Sarah, WINDLE, Gill, CRAIG, Claire and SPRANGE, Kirsty (2017). The long-term (24-month) effect on health and well-being of the Lifestyle Matters community-based intervention in people aged 65 years and over: a qualitative study. BMJ Open, 7 (9), e016711. Copyright and re-use policy See http://shura.shu.ac.uk/information.html See http://shura.shu.ac.uk/information.html ABSTRACT To cite: Chatters R, Roberts J, Mountain G, et al. The long- term (24-month) effect on health and well-being of the Lifestyle Matters community- based intervention in people aged 65 years and over: a qualitative study. BMJ Open 2017;7:e016711. doi:10.1136/ bmjopen-2017-016711 Strengths and limitations of this study Objectives To assess the long-term effect on health and well-being of the Lifestyle Matters programme. ► ►We present data obtained from a purposeful sample of 13 participants who received the Lifestyle Matters intervention within a randomised controlled trial. Design Qualitative study of a subset of intervention arm participants who participated in the Lifestyle Matters randomised controlled trial (RCT). ► ►We analysed the interview data thoroughly using framework analysis. Setting The intervention took place at community venues within two sites in the UK. ► ►Recall may be an issue as participants were asked to recall events that occurred up to 2 years ago. ► ►Recall may be an issue as participants were asked to recall events that occurred up to 2 years ago. Participants A purposeful sample of 13 participants aged between 66 and 88 years from the intervention arm of the RCT were interviewed at 24 months post randomisation. Interviews aimed to understand how participants had used their time in the preceding 2 years and whether the intervention had any impact on their lifestyle choices, participation in meaningful activities and well-being. ► ►Prepublication history for this paper is available online. To view these files, please visit the journal online (http://dx.doi. org/10.1136/bmjopen-2017- 016711). Sheffield Hallam University Research Archive http://shura.shu.ac.uk Sheffield Hallam University Research Archive http://shura.shu.ac.uk group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Research To cite: Chatters R, Roberts J, Mountain G, et al. The long- term (24-month) effect on health and well-being of the Lifestyle Matters community- based intervention in people aged 65 years and over: a qualitative study. BMJ Open 2017;7:e016711. doi:10.1136/ bmjopen-2017-016711 The long-term (24-month) effect on health and well-being of the Lifestyle Matters community-based intervention in people aged 65 years and over: a qualitative study The long-term (24-month) effect on health and well-being of the Lifestyle Matters community-based intervention in people aged 65 years and over: a qualitative study Robin Chatters,1 Jennifer Roberts,2 Gail Mountain,1 Sarah Cook,3 Gill Windle,2 Claire Craig,3 Kirsty Sprange4 Background Older adults can experience many life changing events in later life, such as retire- ment, disability and illness, which may have deleterious effects on mental and physical health and well-being.1 In turn, decline and loss can result in social isolation and lone- liness, which further erodes quality of life. Participation in meaningful activities can prevent this decline.2 3 Occupational therapy aims to help individuals participate in both necessary and meaningful activities; it has an important role to play in health promotion and disease prevention by supporting engage- ment in activity and maintenance of occupa- tional balance.4 Older adults can experience many life changing events in later life, such as retire- ment, disability and illness, which may have deleterious effects on mental and physical health and well-being.1 In turn, decline and loss can result in social isolation and lone- liness, which further erodes quality of life. Participation in meaningful activities can prevent this decline.2 3 Occupational therapy aims to help individuals participate in both necessary and meaningful activities; it has an important role to play in health promotion and disease prevention by supporting engage- ment in activity and maintenance of occupa- tional balance.4 Received 3 March 2017 Revised 10 August 2017 Accepted 16 August 2017 Intervention Lifestyle Matters is a 4-month occupational therapy intervention, consisting of group and individual sessions, designed to enable community living older people to make positive lifestyle choices and participate in new or neglected activities through increasing self- efficacy. Results Interviews revealed that the majority of interviewed participants were reportedly active at 24 months, with daily routines and lifestyles not changing significantly over time. All participants raised some form of benefit from attending Lifestyle Matters, including an improved perspective on life, trying new hobbies and meeting new friends. A number of intervention participants spoke of adapting to their changing circumstances, but there were significant and lasting benefits for 2 of 13 intervention participants interviewed. 1School of Health and Related Research, The University of Sheffield, Sheffield, UK 2Dementia Services Development Centre, Bangor University, Bangor, UK 3Health and Social Care Research Centre, Sheffield Hallam University, Sheffield, UK 4Faculty of Medicine & Health Sciences, Nottingham University, Nottingham, UK Correspondence to Robin Chatters; r.chatters@sheffield.ac.uk ► ►Prepublication history for this paper is available online. To view these files, please visit the journal online (http://dx.doi. org/10.1136/bmjopen-2017- 016711). Received 3 March 2017 Revised 10 August 2017 Accepted 16 August 2017 Open Access Two studies undertaken in the USA (and recom- mended in NICE guidance) have investigated the clinical and cost-effectiveness of the Lifestyle Redesign inter- vention.9 10 This is an occupational therapy intervention designed to support older adults at risk of decline in the participation in meaningful activities, thereby increasing their health and well-being.9 10 Quantitatively, Lifestyle Redesign was found to improve the health and well-being of older adults living in a form of sheltered accommoda- tion in a first study,9 11 and in independently living older adults attending some form of community provision in a second study.10 No qualitative work has been published to describe participants’ experiences of the US interven- tion. Lifestyle Matters was inspired by the US Lifestyle Redesign and has been adapted to the UK context. delivery. Both sets of participants were provided with questionnaires at baseline, 6 and 24 months. The primary outcome was the mental health component of the 36-Item Short Form Health Survey (SF-36) questionnaire, which was not significantly different between the control and intervention groups at either 6 or 24 months.15 16 Participants were also interviewed qualitatively at 6 and 24 months to understand their experience of the inter- vention and any lasting effects. Multi-methods research, where qualitative research is undertaken alongside quantitative methods, can be used in RCTs to explore the reasons for the quantitative findings and to explain variations in the effectiveness of an intervention.17 Medical Research Council Frame- work recommends using qualitative research alongside trials ‘to explain discrepancies between expected and observed outcomes, to understand how context influ- ences outcomes’.18 We therefore undertook a qualitative study 24 months post-randomisation to understand the long-term effects of the Lifestyle Matters intervention in order to ascertain if changes in well-being which were not identified quantitatively could be identified qualitatively. g p Based on an occupational approach to healthy ageing, the manualised Lifestyle Matters intervention is designed to assist participants to improve their well-being and avoid the decline associated with social isolation and poor mental health.12 It involves participants meeting in a weekly group of up to 12 people over 4 months at a local community venue. Participants are also asked to engage in monthly individual sessions with one of the two facil- itators at mutually agreed locations. Methods The reporting of this study conforms to the Consolidated criteria for reporting qualitative research (COREQ) guidelines.19 Open Access Session topics can either be chosen from the manualised programme or new topics identified.12 The facilitators work with the partici- pants to explore the selected topic through discussion, activities and enactment in the community. The emphasis throughout is on the identification of participants' own goals, empowerment through sharing strengths and skills and providing support to enable them to practice new or neglected activities independently, particularly in the community. The intervention was postulated to change behaviour through Bandura’s theory of self-efficacy, which theorises that self-efficacy is developed through four main sources—performance accomplishments (where success undertaking an activity builds self-belief in efficacy), vicarious experiences (seeing people similar to oneself undertake a task successfully), verbal persuasion that one has the ability to succeed in a given activity and inferences from emotional stimuli indicative of personal strengths and vulnerabilities.13 The findings from recent systematic reviews support the use of psychosocial interventions for ageing well. Interventions identified through these reviews include engagement in meaningful social activities,5 those designed to support occupational engagement6 and multicomponent group-based health promo- tion interventions.7 Guidance published by the UK National Institute for Health and Care Excellence (NICE) supports the use of group and individual sessions based on occu- pational therapy principles, where partici- pants rather than facilitators are positioned as experts and peer support is fostered.8 Conclusion The majority of those who experienced the Lifestyle Matters intervention reported minor benefits and increases in self-efficacy, but they did not perceive that it significantly improved their health and well-being. The two participants who had experienced major benefits also reported having had life-changing events, suggesting that this intervention is most effective at the time when lifestyle has to be reconsidered if mental well-being is to be sustained. Correspondence to Robin Chatters; r.chatters@sheffield.ac.uk Trial registration ISRCTN, ISRCTN67209155, post results. Trial registration ISRCTN, ISRCTN67209155, post results Trial registration ISRCTN, ISRCTN67209155, post results. Trial registration ISRCTN, ISRCTN67209155, post results. 1 Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Recruitment and data collection Interviews were undertaken at 24 months postrando- misation (approximately 18 months after intervention delivery had completed) by JR (Female Research Assis- tant with PhD) and RC (Male Trial Manager), both having experience of undertaking interviews with older adults. Participants were recruited to the study through purposeful sampling from the intervention arm of the study. All intervention participants who had been inter- viewed at 6 months and had provided 24-month quan- titative follow-up data were approached to participate once more (n=13). Potential participants were first approached by JR or RC via a telephone call to ascertain initial interest in the study; no other previous contact was had between the researchers and participants prior to this. If interested, a participant information sheet and copy of the consent form was sent by post, and the partic- ipant was telephoned again to confirm that the informa- tion had been read, and he/she was happy to participate. Of those approached, nine participated in the substudy. Participants did not participate due to either not being contactable (n=2), not being interested in participating (n=1) or had already withdrawn consent from the main trial (n=1). Six additional intervention group participants were then approached to reflect a range of health-related experiences and abilities (based on their SF-36 scores at 24 months follow-up, age, location (North Wales/Northern England) number of intervention sessions attended and professional background (skilled, non-skilled) and g We undertook the Lifestyle Matters randomised controlled trial (RCT) to examine the clinical and cost-ef- fectiveness of Lifestyle Matters, the protocol for which is reported elsewhere.14 Between August 2012 and April 2013, 288 individuals living in an urban area in the North of England and 12 rural area in North Wales were recruited to the Lifestyle Matters study via general practitioner (GP) mail outs, community engagement and healthcare professional referrals. Individuals recruited to the study were randomised to either the intervention or usual care, in couples (if both individuals consented to participate in the study) or individually. Participants randomised to the intervention received the manualised 4-month Lifestyle Matters intervention, facilitated by NHS band four staff who were provided with preliminary training and super- vised by qualified occupational therapists throughout Chatters R, et al. BMJ Open 2017;7:e016711. Recruitment and data collection doi:10.1136/bmjopen-2017-016711 2 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Table 1 Demographics of interviewed participants Participant ID Site Age range Gender SF-36 MH score (6 months) SF-36 MH score (24 months) Number of group sessions attended Number of one-to-one sessions received/ number offered P14 Northern England 65–74 Female 60 70 13 1/4 P15 Northern England 65–74 Male 85 65 12 1/4 P16 Northern England 65–74 Female 65 80 14 2/4 P17 Northern England 65–74 Female 90 80 9 0/4 P18 Northern England 85–94 Female 90 70 12 1/4 P25 North Wales 85–94 Male 95 95 13 2/3 P20 North Wales 75–84 Male 70 90 15 1/4 P21 North Wales 65–74 Male 70 90 15 1/4 P22 North Wales 65–74 Male 90 90 11 1/4 P23 North Wales 65–74 Female 90 95 14 3/4 P24 North Wales 75–84 Female 85 100 1 0/0 P19 Northern England 65–74 Male 90 70 15 1/4 P26 North Wales 65–74 Female 85 80 15 1/4 SF-36, 36-Item Short Form Health Survey, scored on a 0 (poor) to 100 (good) health scale. SF-36, 36-Item Short Form Health Survey, scored on a 0 (poor) to 100 (good) health scale. directly about it, no participants mentioned attending any one-to-one sessions in their interviews. gender). Four of these consented, with participants not participating due to not being contactable (n=1) and not being interested in participating (n=1). In total, 13 participants were interviewed at 24 months. Participant demographics are summarised in table 1. The average age of participants was 73.0 years and ranged from 65 to 88 years. Six males were interviewed and six participants lived alone. The average SF-36 score of the sample was 82.7. One participant was in a couple with another study participant (who was not interviewed in this qualitative study). The semi-structured interview schedule aimed to explore, over the previous 18-month period (ie, from the end of the intervention phase): a) any changes in use of time and social participation; b) any impact that Lifestyle Matters on their lives and if so how and why. The interview scheduled was piloted with a patient representative; the data from which was not used in the final analysis. Table 2 presents the questions which partic- ipants were asked, including example prompts. Lifestyle Matters group attendance ranged from 1 to 15 sessions. Interviewed participants represented eight Lifestyle Matters groups, four in each site. Recruitment and data collection The number of individual sessions offered to participants ranged between zero and four, the number actually received ranged between zero and three. Although they were not asked All interviews were conducted in the homes of partic- ipants; no one else was present and repeat interviews were not undertaken. The participants would have been aware of the researcher’s interest in the Lifestyle Matters intervention. Consent to audio record the interview was obtained from all participants and recordings were 3 Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 Table 2 Semi-structured interview questions and example prompts Question Example prompt How has life been for you over the last 18 months? How has life changed for you over the last 18 months? Have things improved or got worse? What does the term ‘quality of life’ mean to you? What would improve the quality of your life? How has life changed for you over the last 18 months? Are you doing more or less activities (social or otherwise) What do you think prevents you from engaging in social/leisure activities, or activities that you used to enjoy doing? Has anything made it more difficult to do such activities? Has the Lifestyle Matters group changed your life in any way? Has the Lifestyle Matters group affected how easy or hard it is for you to engage in social activities? 3 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Open Access in the presence of a researcher (RC or JR) by the partici- pant on the day of the interview, when consent to record the interview was also obtained. Ethical approval for the main trial and this qualitative substudy was granted by the South Yorkshire Research Ethics Committee (reference number 12/YH/0101). Transcripts were anonymised prior to data analysis in order to preserve confidentiality. transcribed verbatim. Data collection was ended when the diversity identified in the sampling frame had been reflected in the interviews. Interviews lasted between 20 and 50 min and field notes were made after the inter- views. Transcripts were not returned to participants for comments or corrections. Mental and physical health Eleven participants reported having either a short-term or long-term health condition that had not changed signifi- cantly over the time period. Three participants discussed how their health had declined due to major illness; two had been receiving treatment for cancer and one partic- ipant’s health had declined due to chronic obstructive pulmonary disease. The intervention did not reportedly have any impact on the physical health of participants in the intervention group. The intervention reportedly had a positive effect on mental well-being. Four participants stated that attending the intervention helped them to ‘gain perspective’ on their lives and what they have. For two of these, the change in perspective helped to broaden their outlook. …it’s … helped me with … knowing what you could do… if there’s a group of us, probably someone comes up with a suggestion or something you know and … it’s probably broadened my outlook a little bit more. (P18, Female, Northern England) …it’s … helped me with … knowing what you could do… if there’s a group of us, probably someone comes up with a suggestion or something you know and … it’s probably broadened my outlook a little bit more. (P18, Female, Northern England) It has made me look at things in a different way. As I said, there are answers to be had, you don’t have to give up, there is a way forward and there are people willing to help you too. (P22, Male, North Wales) The other two participants related the apparent social isolation of fellow participants in the group to their own situation, helping them to appreciate what they have. (Lifestyle Matters has) given me an idea how the other half live, in a way, and I suppose in a way I was very pleased with my … position in life. (P25, Male, North Wales) …it makes me appreciate … my friends and my husband still being with me and the life that I have and my family that I have got… (P24, Female, North Wales) Data analysis A deductive framework approach was used for data anal- ysis,20 the starting point for which was the theory by which the intervention was postulated to function (as described above). Framework Analysis is a five-stage process of: familiarisation, forming a thematic framework, indexing, charting and mapping and interpretation. Familiarisation was achieved through reading and rereading a selection of interview transcripts. A thematic framework, consisting of major themes and one level of minor themes, was developed by two researchers (JR and RC). A small number of transcripts were coded using the framework, after which the researchers met to discuss the framework and finalise any changes that needed to be made. The same two transcripts were coded by three researchers (JR, RC and OO) to ensure comparable coding; any differ- ences were discussed. All other transcripts were coded by one researcher using NVIVO V.10 software. Matrices were then developed, with one theme per matrix. Themes were derived from the data. Matrices were split into activities, attitudes and personal resources, health, mobility, quality of life (overview), social, family, community and other experiences and effects of Lifestyle Matters. These matrix charts were then examined for cross cutting themes and patterns in the data were mapped to inform the final level of interpretation. In order to report the results in a coherent manner, health, mobility and quality of life matrices were combined to form ‘mental and physical health’. Attitudes and personal resources and commu- nity were combined to form ‘personal and community resources’. New matrices were coded using elements of other charts to form the following new major themes: new skills and knowledge, change in routine, new leisure activities, meeting new people, family commitments and bereavement. We attempted to identify if the interven- tion increased self-efficacy through any of Bandura’s four sources of self-efficacy (verbal persuasion, performance accomplishment, emotional arousal and vicarious experi- ence), using the participant’s own words. Participants did not feedback on the findings. Results Table 3 summarises the impact of the intervention on interviewed intervention group participants and the asso- ciated theoretical underpinning. Table 3 summarises the impact of the intervention on interviewed intervention group participants and the asso- ciated theoretical underpinning. Recruitment and data collection Although sample size was prespecified, data saturation was discussed and confirmed following completion of all interviews; transcripts were read and themes identified to ensure that no new themes were arising in the final few interviews. Ethical considerations It should be noted that this latter participant (P24) only attended one intervention session, so it is unlikely that the intervention had an effect on self-efficacy. All participants gave written informed consent for partic- ipation in the study. Participants were telephoned to ascertain their interest in participating, and were sent a consent form and information sheet that explained the purpose of the study. The consent form was then signed Other participants discussed how being involved had improved their mental well-being. Two described how attending the group aspect of the intervention provided Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 4 Open Access Table 3 Impact of the Lifestyle Matters intervention and associated theoretical underpinning Participant ID Benefit(s) Self-efficacy source P14 Started computer lessons and now uses it regularly Performance accomplishment P15 Took up Bridge following support from group Verbal persuasion P16 Confidence and support after death of husband Performance accomplishment P17 Made new friends Joined a keep fit class following support from group Performance accomplishment P18 Broadened outlook Emotional arousal P25 Gave perspective seeing how ‘less fortunate’ people live Emotional arousal P20 Improved confidence to socialise with new people; now socialises more Performance accomplishment/vicarious experience P21 Helped ‘pull out of misery’ following illness Emotional arousal P22 Gained perspective Helped him ‘find answers’ (now using credit union) Emotional arousal P23 Improved confidence in group situations Performance accomplishment/vicarious experience P24 Gained perspective Emotional arousal P19 Increased confidence to undertake new activities Mental lift Positive impact on spousal relationship Performance accomplishment Emotional arousal P26 Undertaking rock climbing with group has provided her with increased confidence to do other things Provided healthy eating information Performance accomplishment group.bmj.com on October 13, 2017 Published by http://bmjopen.bmj.com/ Downloaded from New skills and knowledge them with a ‘mental lift’, which lasted a short period of time after each session. Around half of those interviewed who had received the intervention stated that they had gained new skills and knowledge from attending Lifestyle Matters. This included information about financial matters, healthy eating and services that can help older people. The majority of participants described gaining knowledge about financial matters and local adult education classes, including computers and keep fit. …it give us a buzz which gave us a mental lift which lasted for 2 or 3 days after…it’s when you talk about it that you think yes perhaps I should be joining some activities and I would get that again perhaps. (P19, Male, Northern England) …it give us a buzz which gave us a mental lift which lasted for 2 or 3 days after…it’s when you talk about it that you think yes perhaps I should be joining some activities and I would get that again perhaps. (P19, Male, Northern England) This had positive effect on the participant’s relation- ship with his wife. we talked quite a lot at …about eating and just sort of how we lived generally and I think it did just sort of widen our scopes a little bit… (P26, Female, North Wales) because of what people said at the group made me appreciate how good she is and made me feel quite content, I have a lot to be happy, a lot to be happy and contented about but I probably didn’t realise. (P19, Male, Northern England) hobbies is cooking and that gives me pleasure. (P22, Male, North Wales) hobbies is cooking and that gives me pleasure. (P22, Male, North Wales) one grip away….and to be able to get up there and get down again, I thought was just amazing… I never thought that I would do that and I did it, means that I can do the zip wire if I feel so inclined. (P26, Female, North Wales) Another stopped knitting due to dexterity and eyesight problems. I used to do a lot of knitting I used to knit for the family and I used to sew my own clothes, but I don’t do that anymore because I haven’t got the same dexterity as I used to have and my eyesight is not as good as it was. (P24, Female, North Wales) The intervention provided both participants with a ‘performance accomplishment’, allowing them to succeed in a new hobby or activity in a safe environment. This positive experience built up their belief in their self-efficacy, making it much more likely that they would try the new activity (or other new activities) again. This was also true for other participants who tried computing and attending opera with the group. Learning about new activities as part of Lifestyle Matters meant that some participants developed new leisure pursuits, including computing, keep fit and attending opera with a group. One participant described how participation in the intervention had a major impact on his life. He had been a keen golfer but his health had deteriorated and he had had to give up golf a year before the study commenced. He had continued to attend the social events at the golf club for a period of time but then this had also stopped because he no longer had as much in common with the other members. He reported how he had started to become lonely, and Lifestyle Matters helped him to discover a new interest. Three participants had discovered a new hobby through participation in the intervention and were still continuing the hobby at the time of their interview. One of these participants met new friends through the inter- vention and joined a keep fit class. Making a new friend appeared to provide a performance accomplishment for this participant. You meet one person and then through that person you meet their friends… then you… join up with them and make friends then you start doing things with them. hobbies is cooking and that gives me pleasure. (P22, Male, North Wales) (P17, Female, Northern England) Bridge seemed like a good idea, but I’d never got off my backside and done anything about it, and the two girls that took the course were very good at kicking me up the backside and making sure I did do something about it, for which I’ve been very grateful as that’s made a huge difference to my life. (P15, Male, Northern England) Eight participants did not describe getting involved in new activities. They said that they already led busy lives and therefore did not have the time or impetus to continue with new activities. Two participants had pursued and enjoyed new activ- ities during group sessions, but did not continue these after the intervention ended. One of which tried and enjoyed crazy golf but did not continue with it; trying the new activity provided him with confidence to try other new activities in the future. The intervention facilitators provided this participant with verbal persuasion in order to help him increase his confidence to initiate a new activity. Adaptation to leisure activities Another participant spoke of how the intervention helped to pull him out of misery following illness. Even though, for the majority of participants, their health did not decline over the time period, physical health affected the activities participants were able to undertake. Participants had to stop or reduce certain activities due to their health. One participant used to walk regularly, but due to physical health/mobility limitations has instead started a new hobby (cooking), although this was not instigated by the Lifestyle Matters intervention. Yes I was still having to go for outpatients appointments… but when you are sort of with a group and … it’s a specific day you are motivated and I think … talking with the group and laughing and joking sort of spurs you on…you know pulls you out of your misery like. (P21, Male, North Wales). The improvement of these participant’s mental state, and the ‘gaining of perspective’ of other participants may have impacted on their somatic and emotional status, allowing participants to interpret stress reactions more positively and therefore possibly increasing their self-efficacy. I use to enjoy going to the country and going for a walk, I’ve had to cut down on the (walking) that would help me…. [I’ve cut down because of] my knee, my back, my physical health. One of my main Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 5 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Personal and community resources Availability of community resources reportedly affected the ability of some participants to undertake activities outside the home; four participants stated that they had been unable to continue to attend classes or the gym because of various reasons such as gym closure, a class having too few members to be viable any longer and death of the person who ran a class. One participant was still meeting on a one-to-one basis with a fellow member of the Lifestyle Matters group, either talking by telephone or undertaking activities together, and subsequently making more friends. just by going to that health thing that we went to up there through this, that [Lifestyle Matters] group, I made some friends there that we still go out with, a couple of ladies and then a walking group we joined. (P17, Female, Northern England) I have not been able to do as much Thai Chi because I used to be able to do it 3 times a week and then because [instructor] didn’t get many people going on a Wednesday he stopped doing it. (P14, Female, Northern England). Meeting new people in a group situation provided two participants with self-efficacy via either performance accomplishment and/or vicarious experience; confidence was gained to interact with people in such situations. The participants who mentioned these challenges were already either attending several other classes, or were able to find a new gym, highlighting their desire to remain proactive. Lifestyle Matters aided them to locate new activities, with the participant who had to stop Thai Chi taking up computer classes instead; the facilitators assisted her to experience performance accomplishment, which resulted in her attending the class again Participant: "I’ve got more confidence now in myself then I used to have years ago yeah". Participant: "I’ve got more confidence now in myself then I used to have years ago yeah". Interviewer: "…do you think that’s anything to do with the groups…" Participant: "It just makes me talk a bit more perhaps". (P23, Female, North Wales) [The facilitator] took us down [to the computer class] and introduced us…we have been ever since and we go every week. (P14, Female, Northern England). Personal and community resources It might have been [the Lifestyle Matters group] … might have given me more confidence to walk into a room full of women… (P20, Male, North Wales) It might have been [the Lifestyle Matters group] … might have given me more confidence to walk into a room full of women… (P20, Male, North Wales) Open Access grandchildren poorly so that has gone a bit haywire at the moment. (P16, Female, Northern England) grandchildren poorly so that has gone a bit haywire at the moment. (P16, Female, Northern England) Further group meetings also stopped after a few weeks or months because people started to drop out. We met up for, I think we were going to meet up one time and then everybody started going on holiday so it weren’t possible but and I think it has just filtered off as I haven’t seen them. (P16, Female, Northern England) None of those interviewed described any impact on how they negotiated family commitments that could be attributed to the intervention. Meeting new people doi:10.1136/bmjopen-2017-016711 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Meeting new people All participants met new people as part of the Lifestyle Matters groups, but only a minority made ‘meaningful’ relationships that lasted past the end of the group. Around half of the interviewed intervention partici- pants stated that they had not continued meeting with the group as they had not met people with similar interests. Some of the activities that we took part in be it crazy golf, visiting museums I did intend to go back and have a go at some of these activities but I have slipped back into my old… my own comfortable lifestyle. (P19, Male, Northern England) relationships that lasted past the end of the group. Around half of the interviewed intervention partici- pants stated that they had not continued meeting with the group as they had not met people with similar interests. Attending the group was good in [the] respect that I know there is things out there and if I want to go and have a try I can go and do it. (P19, Male, Northern England) …although the people were all right they were nice people there was nobody really that I would have wanted to have kept on with you know (P14, Female, Northern England) The second mentioned enjoying climbing during the intervention. She had not continued with this after the intervention had finished, but doing so had increased her confidence; as a result, she had the confidence to under- take an activity that she never thought were possible. Six participants had plans to meet up with their fellow group members after the intervention had finished, but this only lasted for a short period of time, as the person who volunteered to do the organising had not been in touch. It’s certainly given me the inclination to do the zip wire. Because we did the … wall climbing. I felt that doing that was, wow, it was just something that I never thought that I would be able to do. And the fact that I practically reached the top of the wall, I was only One of the members had promised to contact each one of us, he had all our telephone numbers, but he didn’t do anything. (P17, Male, North Wales) I practically reached the top of the wall, I was only 6 Chatters R, et al. BMJ Open 2017;7:e016711. Discussion This qualitative study found that those who had partici- pated in Lifestyle Matters reportedly enjoyed the inter- vention and all spoke of receiving some form of benefit but only 2 out of the 13 we interviewed were able to describe any major long-term impact. Many participants described a process of adapting the type of activity they undertook following changes in their circumstance—two participants swapped active hobbies for sedentary ones (e.g., walking for cooking) following mental or physical decline; other participants, following a change in commu- nity resources, had to find new activities. A few partici- pants had to adapt their activities following bereavement of their partner. Participants displayed self-efficacy to adapt to their new circumstances, although only two of the interviewed intervention participants were able to attri- bute this adaptability to the intervention. It is significant therefore that these two individuals had recently experi- enced a challenge in their lives immediately prior to their engagement in the intervention; one had been recently bereaved and the other had recently had to adjust to a reduction in mobility. This finding indicates that the Life- style Matters intervention was most meaningful for those at a ‘trigger point’ in their life which demanded transi- tion and change. The intervention assisted those in the intervention arm who were at this trigger point to adapt to their new circumstances. Lifestyle Matters provided sign-posting to new activities for those individuals whose current activities were no longer available. Self-efficacy was boosted by providing participants with performance accomplishment (where participants succeeded at new activities) and modelling influences (where participants witness their peers undertake activities successfully).13 Emotional arousal was provided by improving mental health and participant’s outlook on life. It is clear that for two participants, the intervention assisted an increase in self-efficacy, which resulted in change in behaviour and increases in quality of life. For other intervention g This qualitative study highlights that in the Lifestyle Matters study, only a small number of participants were at a ‘trigger point’. Open Access when you end up on your own you don’t go out of these four walls I would think you can lose that confidence and very quickly. (P16, Female, Northern England) participants, there are examples described above of their self-efficacy increasing, but this did not result in a major change in their day-to-day activities and resulting quality of life. It is evident that barriers were present which prevented participants from adapting—barriers included family commitments, physical and mental health, commu- nity resources and bereavement. Attending Lifestyle Matters helped her to regain her confidence and re-engage in social activities. This study has a number of strengths. Interviews were undertaken and analysed by two researchers, ensuring that objectivity was maximised. It should be noted that these interviews were undertaken during the winter at a time of prolonged snow and icy conditions; such weather conditions have previously been shown to reduce phys- ical activity levels in older adults with self-reported limita- tions in activities of daily living.21 Participants were asked to discuss life events covering a 2-year period. Recall may have also been an issue, especially when discussing the impact of an intervention they received 2 years previously. Participants experience of the intervention and resulting increases in self-efficacy were linked to Bandura’s four sources of self-efficacy through analysing participant’s experience of the intervention and associated benefits. It was difficult to determine if possible increases in self-ef- ficacy (e.g., increased confidence) were due to vicarious experience or mastery experiences, as participants did not discuss explicitly which aspect of the intervention led to such changes. I think it gives you something some kind of confidence when you when you know you do end up on your own and err and I just think it gives you confidence because to meet people and to be in a group.(P16, Female, Northern England) It is clear that the intervention provided this participant with a ‘performance accomplishment’—she was nervous about going alone to a new group of strangers, but having succeeded to make friends, she has more confidence to undertake such an activity again. Bereavement doi:10.1136/bmjopen-2017-016711 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Bereavement Family commitments influenced participant’s use of time, with three interviewees reporting that family was a major part of their routine. One of those participants reported an increased commitment to taking care of family because he had recently started taking care of his grandchildren full time, which had resulted in a change in use of time: Bereavement was mentioned by a number of participants. Death of a spouse, family member or friend had a marked effect on lifestyle. All participants who had lost a partner spoke of having to adapt and develop in some way, including developing new skills or re invigorating neglected ones and engaging in new social activities. Apart from looking after the little girls full time, that’s changed my life in the past eighteen months or so. This has restricted my personal activities a lot. So I sometimes feel that I don’t get enough time to do what I want to do. (P17, Male, North Wales) …when you make new friends when you lose the friends that you have got if their still with their husbands it is, I don’t know it makes you feel a bit, its hard to put into words sometimes [pauses] [cries] it is an emptiness and you have got to try and fill this emptiness yes [pause] the only way to do it is to, this is me, is to make myself do things and go out and speak to people. (P16, Female, Northern England) A frequently described situation was that of having to undertake routines to fit in with the work-life routines of family, specifically in order to take care of grandchildren. One participant commented that she did not feel the balance was currently in favour of her personal interests: Engagement in the intervention reportedly had a major impact on this participant’s life. This woman’s husband had died prior to her involvement in Lifestyle Matters. Participation reportedly helped her to deal with the recent bereavement which had left her low in confidence and with fewer social opportunities. because family when they are working and that you seem to get set days and that can you know so you end up doing that (….) whether you want to or not yeah you know they rely on you so you feel obliged to do it….At the moment no because I have had some of my 7 Chatters R, et al. BMJ Open 2017;7:e016711. References 1. Mountain GA, Craig CL. The lived experience of redesigning lifestyle post-retirement in the UK. Occup Ther Int 2011;18:48–58. 2. Kidd JM. Foresight Mental Capital and Wellbeing: Making the most of ourselves in the 21st century: Government Office for Science, 2008. http://www.bis.gov.uk/assets/biscore/corporate/migratedD/ec_ group/116-08-FO_b 3. Age Concern and, Mental Health Foundation. Promoting mental health and well-being in later life. A first report from the UK inquiry into mental health and well-being in later life. 2006. 4. Scaffa ME, Van Slyke N, Brownson CA. American Occupational Therapy Association Commission on Practice. Occupational therapy services in the promotion of health and the prevention of disease and disability. Am J Occup Ther 2008;62:694–703. 5. Forsman AK, Nordmyr J, Wahlbeck K. Psychosocial interventions for the promotion of mental health and the prevention of depression among older adults. Health Promot Int 2011;26:i85–i107. 6. Arbesman M, Mosley LJ. Systematic review of occupation- and activity-based health management and maintenance interventions for community-dwelling older adults. Am J Occup Ther 2012;66:277–83. 7. Windle G, Hughes D, Linck P, et al. Public health interventions to promote mental well-being in people aged 65 and over: systematic review of effectiveness and cost-effectiveness. NICE 2008. Future research needs to be undertaken in order to identify how isolated older adults can be identified and recruited to preventative health interventions such as Lifestyle Matters. The fact that only 2 out of 13 interven- tion participants gained ‘major’ benefit from the inter- vention could be due to the recruitment methods used in this study, where participants were recruited, in the majority, via GP mail outs, resulting in a physically and mentally well population. The majority of participants who were interviewed in this study were not at a ‘trigger point’ in their life when they were in need of a health and well-being intervention such as Lifestyle Matters. Despite the emphasis current guidance places on recruiting isolated older adults to preventative lifestyle interven- tions, other studies have also discussed issues with iden- tifying such individuals.8 26 27 Future evaluations should be undertaken to assess the viability of other recruitment techniques, such as respondent-driven sampling, which has been used previously to recruit isolated adults with HIV.28 8. National Institute for Health and Care Excellence (NICE). Mental wellbeing in over 65s: occupational therapy and physical activity interventions. London. 2008. https://www.nice.org.uk/guidance/ ph16. p 9. Clark F. Occupational therapy for independent-living older adults. A randomized controlled trial. JAMA 1997;278:1321–6. 10. Discussion This is in contrast to the US Lifestyle Redesign studies, and the UK feasibility study which was undertaken prior to the RCT, which suggest that all partic- ipants gained from the intervention in a meaningful way when assessed in the USA10 22 and in the UK.1 However, this difference may be explained by the lack of long-term qualitative follow-up in these preceding studies; qualita- tive work was not undertaken in the US studies, and the UK feasibility study qualitatively followed up participants directly after the cessation of the facilitated group sessions and therefore did not assess the long-term impact of the intervention. The quantitative and qualitative findings from this work did not demonstrate that the intervention had any impact on quality of life at population level. The majority of those recruited to participate reported being busy, active and resourceful, engaged in a diverse range of activities and with established social networks and it can therefore be argued that they had little to gain from the intervention. Inability to identify those at risk of mental decline appears to be a prime reason for the findings obtained through both quantitative and qualitative enquiry. Another issue that may have contributed includes the duration of the intervention (4 months), which was far shorter than for the US Lifestyle Redesign studies (6 months duration) and for the Lifestyle Matters feasibility study (8 months duration).10 22 A longer delivery duration can give those with potentially ‘hidden’ challenges the time to become sufficiently confident to work with others to explore Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 8 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Open Access Ethics approval Ethical approval for the main trial and this qualitative substudy was granted by the South Yorkshire Research Ethics Committee (reference number 12/YH/0101). All participants consented to participate in this qualitative study. solutions. In addition, it can be argued that implicit issues such as social desirability and unacceptable disease bias,23 24 which have been identified in other qualitative studies,25 may mean that the benefit of the intervention was under-reported. One of the aims of the group sessions was to aid participants in moving forward together with all parties taking an active role in organising outings and meet ups. Discussion Unfortunately, the majority of participants did not continue in their groups after the cessation of the intervention, often because the person who volunteered to continue to organise the groups did not make contact with the other group members. Provenance and peer review Not commissioned; externally peer reviewed. Data sharing statement All relevant data are published within this manuscript. Open Access This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 4.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/ licenses/by/4.0/ © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted. Clinicians and policy makers should consider who is best to facilitate an intervention such as Lifestyle Matters. In the current study, the facilitators were NHS band four therapists, who did not have in-depth training in how to manage such groups (although they did receive study-specific training). More highly trained facilitators, such as occupational therapists, may have improved the delivery of the intervention. In addition, the Lifestyle Matters intervention could be further developed in order to improve the benefit to participants. Although they were not asked specifically, none of the interviewed participants mentioned the one-to-one sessions in their interview or stated they had gained any benefit from them; the intervention could therefore be further devel- oped by increasing the understanding of the purpose of the one-to-one sessions with both the intervention partic- ipants and facilitators. References Clark F, Jackson J, Carlson M, et al. Effectiveness of a lifestyle intervention in promoting the well-being of independently living older people: results of the Well Elderly 2 Randomised Controlled Trial. J Epidemiol Community Health 2012;66:782–90. 11. Clark F, Azen SP, Carlson M, et al. Embedding health-promoting changes into the daily lives of independent-living older adults: long- term follow-up of occupational therapy intervention. J Gerontol B Psychol Sci Soc Sci 2001;56:P60–P63. y 12. Craig C, Mountain GA. Lifestyle matters: An occupational approach to healthy ageing. Speechmark 2007.i 13. Bandura A. Self-efficacy mechanism in human agency. Am Psychol 1982;37:122–47. 14. Sprange K, Mountain GA, Brazier J, et al. Lifestyle Matters for maintenance of health and wellbeing in people aged 65 years and over: study protocol for a randomised controlled trial. Trials 2013;14:302–13. ; 15. Syddall HE, Martin HJ, Harwood RH, et al. The SF-36: a simple, effective measure of mobility-disability for epidemiological studies. J Nutr Health Aging 2009;13:57–62. g g 16. Mountain G, Windle G, Hind D, et al. A preventative lifestyle intervention for older adults (lifestyle matters): a randomised controlled trial. Age Ageing 2017:627–34. Contributors RC, KS, JR, SC, CC, GM and GW designed the study. RC and JR undertook data collection. RC, JR and SC analysed the data. RC and JR wrote the first draft of the manuscript; all authors reviewed and approved the manuscript. g g g 17. Lewin S, Glenton C, Oxman AD. Use of qualitative methods alongside randomised controlled trials of complex healthcare interventions: methodological study. BMJ 2009;339:b3496. g y ; 18. Craig P, Dieppe P, Macintyre S, et al. Developing and evaluating complex interventions: the new Medical Research Council guidance. BMJ 2008;337:a1655. Funding This work was supported by the Medical Research Council grant number (G1001406). Funding This work was supported by the Medical Research Council grant number (G1001406). 19. Tong A, Sainsbury P, Craig J. Consolidated criteria for reporting qualitative research (COREQ): a 32-item checklist for interviews and focus groups. Int J Qual Health Care 2007;19:349–57. Competing interests None declared. 9 Chatters R, et al. BMJ Open 2017;7:e016711. doi:10.1136/bmjopen-2017-016711 group.bmj.com on October 13, 2017 - Published by http://bmjopen.bmj.com/ Downloaded from Open Access Open Access 24. Nederhof AJ. Methods of coping with social desirability bias: A review. Eur J Soc Psychol 1985;15:263–80. 24. Nederhof AJ. Methods of coping with social desirability bias: A review. Eur J Soc Psychol 1985;15:263–80. 20. Ritchie J, Spencer L. Qualitative data analysis for applied policy research. The Qualitative Researcher’s Companion: SAGE Publications, 2002:305–29. http://books.google.hu/books?hl=de&l r=&id=46jfwR6y5joC&oi=fnd&pg=PA305&dq=policy+research+metho dology&ots=smCUJOpwRT&sig=TAbCw7dKNpsEojF1SyiH0J-3zlE& redir_esc=y#v=onepage&q=policy research methodology&f=false y 25. Ismail H, Atkin K. The NHS Health Check programme: insights from a qualitative study of patients. Health Expect 2016;19. 25. Ismail H, Atkin K. The NHS Health Check programme: insights from a qualitative study of patients. Health Expect 2016;19. 26. Stineman MG, Strumpf N, Kurichi JE, et al. Attempts to reach the oldest and frailest: recruitment, adherence, and retention of urban elderly persons to a falls reduction exercise program. Gerontologist 2011;51:S59–S72. 21. Sumukadas D, Witham M, Struthers A, et al. Day length and weather conditions profoundly affect physical activity levels in older functionally impaired people. J Epidemiol Community Health 2009;63:305–9. 27. Harris R, Dyson E. Recruitment of frail older people to research: lessons learnt through experience. J Adv Nurs 2001;36:643–51 . 22. Clark F, Azen SP, Zemke R, et al. Occupational therapy for independent-living older adults. A randomized controlled trial. JAMA 1997;278:1321–6. 28. Wylie JL, Jolly AM. Understanding recruitment: outcomes associated with alternate methods for seed selection in respondent driven sampling. BMC Med Res Methodol 2013;13:93. 23. Sackett DL. Bias in analytic research. J Chronic Dis 1979;32:51–63. 23. Sackett DL. Bias in analytic research. J Chronic Dis 1979;32:51–63. 10 Chatters R, et al. 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https://openalex.org/W2302509689	https://eprints.gla.ac.uk/117264/7/117264.pdf	English	null	A dynamic organic Rankine cycle using a zeotropic mixture as the working fluid with composition tuning to match changing ambient conditions	Applied energy	2,016	cc-by	11,553	a r t i c l e i n f o Article history: Received 21 December 2015 Received in revised form 16 February 2016 Accepted 6 March 2016 Keywords: Organic Rankine cycle Zeotropic mixture Dynamic composition tuning Distillation Economic analysis Article history: Received 21 December 2015 Received in revised form 16 February 2016 Accepted 6 March 2016 Air-cooled condensers are widely used for Organic Rankine Cycle (ORC) power plants where cooling water is unavailable or too costly, but they are then vulnerable to changing ambient air temperatures especially in continental climates, where the air temperature difference between winter and summer can be over 40 C. A conventional ORC system using a single component working ﬂuid has to be designed according to the maximum air temperature in summer and thus operates far from optimal design condi- tions for most of the year, leading to low annual average efﬁciencies. This research proposes a novel dynamic ORC that uses a binary zeotropic mixture as the working ﬂuid, with mechanisms in place to adjust the mixture composition dynamically during operation in response to changing heat sink condi- tions, signiﬁcantly improving the overall efﬁciency of the plant. The working principle of the dynamic ORC concept is analysed. The case study results show that the annual average thermal efﬁciency can be improved by up to 23% over a conventional ORC when the heat source is 100 C, while the evaluated increase of the capital cost is less than 7%. The dynamic ORC power plants are particularly attractive for low temperature applications, delivering shorter payback periods compared to conventional ORC systems. Keywords: Organic Rankine cycle Zeotropic mixture Dynamic composition tuning Distillation Economic analysis 2016 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/). h i g h l i g h t s A dynamic ORC using a zeotropic mixture with composition tuning is proposed. The working principle is veriﬁed theoretically, based on a thermodynamic model. Improvements in the resultant power plant’s annual power production are analysed The economic beneﬁts have been demonstrated by an economic analysis. A dynamic organic Rankine cycle using a zeotropic mixture as the working ﬂuid with composition tuning to match changing ambient conditions Peter Collings, Zhibin Yu ⇑, Enhua Wang Peter Collings, Zhibin Yu ⇑, Enhua Wang Systems, Power & Energy Research Division, School of Engineering, University of Glasgow, Glasgow G12 8QQ, UK Applied Energy 171 (2016) 581–591 Applied Energy 171 (2016) 581–591 ⇑Corresponding author. Tel.: +44 0 131 440 2530. E-mail address: Zhibin.Yu@glasgow.ac.uk (Z. Yu). http://dx.doi.org/10.1016/j.apenergy.2016.03.014 0306-2619/ 2016 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). emission estimate http://dx.doi.org/10.1016/j.apenergy.2016.03.014 0306-2619/ 2016 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). ⇑Corresponding author. Tel.: +44 0 131 440 2530. E-mail address: Zhibin.Yu@glasgow.ac.uk (Z. Yu). 1. Introduction mance of some organic Rankine cycle systems using zeotropic mix- tures as working ﬂuid for power generation from low-temperature geothermal water, and found that the maximum thermal efﬁciency can reach 9.01% when the heat source temperature is about 100 C [7]. Liu et al. analysed the performance of a geothermal organic Rankine cycle using an R600a/R601a mixture as working ﬂuid, and found that it could generate 11% more power than an ORC using pure R600a when the geothermal water temperature is 110 C [8]. [ ] So far, most ORC systems have considered single-pure compo- nent organic working ﬂuids [9,10]. The heat transfer efﬁciency of the evaporator of an ORC is very important. Pinch point tempera- ture difference is often used to analyse the coupling heat transfer in the evaporator. Chen et al. proposed a method to optimise the operating parameters of an ORC with constrained inlet tempera- ture of the heat source, and the pinch point temperature difference in the evaporator [11]. Normally, there is always a mismatch in the temperature proﬁles between heat transfer ﬂuid and the working ﬂuid because the pure working ﬂuid changes phase at a constant temperature while the heat transfer ﬂuid changes temperature during heat transfer. According to Yu et al., there exists a linear relationship between the integrated-average temperature differ- ence and the exergy destruction in heat exchangers [12]. The integrated-average temperature differences of pure working ﬂuids are large due to the constant temperature during phase change, increasing irreversibility and reducing the cycle efﬁciency [13]. Zeotropic mixtures with temperature varying during phase change can match the temperature proﬁles of the heat source and heat sink better. Aghahosseini and Dincer compared the performances of a low-grade heat source for different pure and zeotropic mixture working ﬂuids [14]. Chen et al. analysed a supercritical simple ORC using zeotropic mixture working ﬂuids and found that an improve- ment of the thermal efﬁciency can be 10–30% compared to a sim- ple ORC using a pure working ﬂuid [15]. Lecompte et al. examined the thermodynamic performance of a non-superheated subcritical ORC with zeotropic mixtures as working ﬂuids and found that an increase in exergy efﬁciency in the range of 7.1–14.2% can be obtained compared to pure working ﬂuids [16]. Braimakis et al. 1. Introduction the normal Rankine cycle, differing only in its use of organic com- pounds as working ﬂuids, is a feasible solution for the utilisation of such low-grade heat sources. Bianchi and De Pascale investigated different energy systems to convert industrial waste energy to electricity and found that the organic Rankine cycle has the highest thermal efﬁciency [2]. Yang et al. studied the performance of an ORC for engine exhaust heat recovery and found the thermal efﬁ- ciency can reach 8.5% [3]. The International Energy Outlook 2013 (IEO2013) forecasted that world energy consumption will grow by 56% between 2010 and 2040 [1]. On the other hand, it is estimated that about 20– 50% of industrial energy input is discharged as waste heat in the form of hot exhaust gases, cooling water, and heat lost from hot equipment surfaces and heated products. To cut greenhouse gas emissions and increase energy efﬁciency, it is critical to recover energy from these low-grade waste heat sources. The Organic Rankine Cycle (ORC), which has the same working principle as Organic Rankine cycles can also be used for other low-grade heat sources such as geothermal energy [4]. 70% of the global geothermal resource is estimated to exist at temperatures of 100–130 C [5]. Geothermal energy takes advantage of zero carbon emissions and the sustainable technical potential for Europe is estimated as 350 TW h/yr [6]. Habka and Ajib assessed the perfor- http://dx.doi.org/10.1016/j.apenergy.2016.03.014 0306-2619/ 2016 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). P. Collings et al. / Applied Energy 171 (2016) 581–591 582 options are limited by the availability of suitable organic ﬂuids. Meanwhile, air-cooled condensers are widely used for ORC power plants where cooling water is unavailable or too costly, but they are then vulnerable to changing ambient air temperatures. For instance, the air temperature difference between winter and sum- mer can reach 30 C in the UK, and can be much higher in locations with continental climates such as Xi’an (China), Warsaw (Poland), and Chicago (US) as shown in Fig. 1. Thus, an ORC power plant designed for a given air temperature (usually the maximum air temperature in summer) operates far from optimal design condi- tions for most of the year, leading to low annual average efﬁcien- cies. This has been identiﬁed in literature as a problem facing Organic Rankine Cycles throughout the world [19]. 1. Introduction studied subcritical and supercritical ORCs based on natural hydro- carbons and their binary mixtures and found that the maximum exergetic efﬁciencies range from 15% to 40% for heat source tem- peratures between 150 and 300 C [17]. Some of the current ORC systems use multi-component work- ing ﬂuids, but none of them can adjust its composition dynami- cally. When the ambient temperature varies, these ORC systems either operate at ﬁxed condensation temperatures or have to reg- ulate the mass ﬂow rate of the organic working ﬂuid. However, only regulating the mass ﬂow rate to improve the system perfor- mance has limited effectiveness. To address this challenge and ﬁll the knowledge gap, this research proposes a novel dynamic organic Rankine cycle that uses a binary zeotropic mixture as the working ﬂuid and can adjust its mixture composition during operation to match the power cycle with the changing ambient air temperature. A higher annual average thermal efﬁciency can be potentially achieved by means of the dynamic ORC. To the best of our knowledge, there currently exist no ORC sys- tems that can dynamically tune their working ﬂuids to match such changing ambient conditions during their operation. In this paper, ﬁrstly, the working principle of a dynamic organic Rankine cycle is developed and analysed theoretically. A numerical model is then established and the thermodynamic beneﬁts are analysed. Finally, through a case study, an economic analysis for the resultant dynamic ORC is conducted and compared with a conventional ORC. The results show that the proposed dynamic ORC can improve the power plant’s annual power production signiﬁcantly while the capital cost is only increased slightly. 2. Dynamic ORC concept When approaching the hottest day in the summer, the low boiling point component R134a will be mostly replaced by the high boiling point component R245fa to have a higher condensing temperature. The resultant cycle is shown as Fig. 3(c). dew line Bubble line Two-phase region Tglide Tdew Tbubble Fig. 2. Dew and bubble lines of a zeotropic mixture of R245fa and R134a at a pressure of 2.9 bar. These process are not a one off adjustment but will be carried out with many small steps according to the ambient temperature change as shown in Fig. 1. The process will be reversed as summer gives way to winter. A similar approach can in theory be adopted on a shorter timescale to maximise efﬁciency over the day–night cycle. Fig. 2. Dew and bubble lines of a zeotropic mixture of R245fa and R134a at a pressure of 2.9 bar. 11 K is the temperature ‘glide’ of this mixture, which can be used to match the temperature change of heat transfer ﬂuid in counter ﬂow heat exchangers, so that the irreversibility can be reduced [23–25]. The use of suitable zeotropic mixtures as working ﬂuids can increase cycle efﬁciency and power production of ORC systems, especially for lower temperature applications (<250 C) [25]. Sec- ondly, zeotropic mixtures display bubble- and dew-point temper- atures between those of the two component ﬂuids, and this varies predictably with mixture percentage. This allows the forma- tion of a mixture with a particular composition to match heat sink temperatures so that more options of working ﬂuids are available [26]. Thirdly, and most interestingly, it is possible to change the composition of the mixture during operation to dynamically match the changing heat source and/or sink temperatures. When the heat source and heat sink temperature, TH and TL, are given, the Carnot cycle efﬁciency is deﬁned as gCarnot ¼ TH TL TH ð1Þ ð1Þ In general, the thermal efﬁciency of an ORC is proportional to the Carnot cycle efﬁciency for the given pair of heat source and heat sink temperatures. The higher the differential (TH TL), the higher the thermal efﬁciency for a given TH. Therefore, a low heat sink tem- perature TL is desirable. As shown in Fig. 4(a), the heat source temperature is ﬁxed as TH, while the ambient temperature varies over the year with the max- imum in summer and the minimum in the winter. 2. Dynamic ORC concept It is proposed here that the challenges outlined in the introduc- tion can be addressed by using zeotropic mixtures, which have the following advantages: Firstly, a zeotropic mixture has a variable temperature during phase change. For example, Fig. 2 shows the bubble and dew lines of a zeotropic mixture formed by R245fa and R134a, generated using REFPROP 9.1 [21]. This mixture has previously been investigated in heat transfer research [22]. For a mixture with 70% of low-boiling-point component R134a, the evaporation starts at the temperature Tbubble around 279 K and ends at Tdew around 290 K. The differential (Tbubble Tdew) around An ORC power plant is currently designed by selecting a work- ing ﬂuid to match the heat source and sink temperatures (i.e., TH and TL). Xu and Yu’s research indicated that the thermal efﬁciency of a subcritical ORC is related to the critical temperature of the working ﬂuid, and recommended organic ﬂuids with critical tem- peratures in the range from 20 to 30 K below the heat source tem- perature to 100 K above it [18]. Considering the heat source and sink temperatures differ from one customer to another, the design -10.00 -5.00 0.00 5.00 10.00 15.00 20.00 25.00 30.00 Jan Feb Mar Apr Apr May Jun Jul Aug Sep Oct Nov Dec Ambient Temperature (°C) Xi'an Warsaw Chicago Fig. 1. Average monthly temperatures of three locations with continental climates: Xi’an (China), Warsaw (Poland) and Chicago (US) [20]. Fig. 1. Average monthly temperatures of three locations with continental climates: Xi’an (China), Warsaw (Poland) and Chicago (US) [20]. P. Collings et al. / Applied Energy 171 (2016) 581–591 583 gradually increases during the seasonal shift from winter to sum- mer, the mass fraction of R134a can be gradually decreased, thus increasing the condensing temperature of the mixture to match the ambient air temperature. As the condensing temperature increases, the evaporating temperature also increases accordingly. In order to ensure the dew point of the mixture is far enough below the heat source temperature to achieve complete evaporation, the evaporating temperature of the mixture can be decreased by reducing the pressure within the evaporator (i.e., reducing both the pump speed and electric load at the generator). After these adjustments, the new cycle is now shown as Fig. 3(b). 2. Dynamic ORC concept In order to demonstrate the performance improvement using dynamic ORC quantitatively, a traditional ORC without composition tuning is selected as a comparative case. Currently, most practical ORC sys- tems are designed based on a ﬁxed condensation temperature that is determined by the designed condensation pressure and the properties of the working ﬂuid. To avoid any leakage of air into the system, the condensation pressure is normally greater than atmospheric pressure. Therefore, the traditional ORC operating with a single working ﬂuid has to be designed according to the maximum air temperature in summer to ensure the working ﬂuid will be fully condensed in the condenser throughout the year. As the season shifts from summer to winter, the ambient temperature decreases. The working ﬂuid in the condenser will be subcooled due to the excessive cooling provided by the colder ambient air This research develops a novel dynamic ORC concept by intro- ducing a composition control mechanism to an ORC operating with a binary zeotropic mixture as the working ﬂuid so that the mixture composition can be dynamically altered during operation to match changing heat sink temperature TL, ensuring higher efﬁciency. Such a dynamic composition control has been proposed and analysed for a heat pump system and showed promising results [27]. However, it has never been considered previously for ORC systems according to a comprehensive literature survey and patent search. Such a Dynamic ORC power plant with an air cooled condenser could be designed and optimised to match the minimum local air temperature in winter as shown in Fig. 3(a). Referring to Fig. 2, the zeotrope will be selected with a high mass fraction of R134a to have a low condensing temperature. As air temperature 250 270 290 310 330 350 370 390 410 430 900 1400 1900 Temperature (K) (a) 900 1400 1900 Entropy (J/kgK) (b) 900 1400 1900 (c) Fig. 3. Change in cycle as the composition of the working ﬂuid is tuned to a changing heat sink temperature, while the heat source temperature remains the same (a) 100% R134a, (b) a 50%:50% mixture of R245fa and R134a, and (c) 100% R245fa. 250 270 290 310 330 350 370 390 410 430 900 1400 1900 Temperature (K) (a) 900 1400 1900 Entropy (J/kgK) (b) 900 1400 1900 (c) Fig. 3. 2. Dynamic ORC concept Change in cycle as the composition of the working ﬂuid is tuned to a changing heat sink temperature, while the heat source temperature remains the same (a) 100% R134a, (b) a 50%:50% mixture of R245fa and R134a, and (c) 100% R245fa. P. Collings et al. / Applied Energy 171 (2016) 581–591 584 Fig. 4. The comparison between the conventional ORC and the proposed dynamic ORC. match the speciﬁc ambient temperatures. In addition, the turbine is assumed to operate with a constant isentropic efﬁciency across the range of pressure ratios presented to it, and this assumption is believed to be feasible. In fact, the research from Pierobon et al. [28] suggests that it is possible to maintain a relatively con- i i bi fﬁi d i i Fig. 4. The comparison between the conventional ORC and the proposed dynamic ORC. Fig. 5. Schematic diagram of a dynamic ORC power plant. Fig. 5. Schematic diagram of a dynamic ORC power plant. Fig. 5. Schematic diagram of a dynamic ORC power plant. match the speciﬁc ambient temperatures. In addition, the turbine is assumed to operate with a constant isentropic efﬁciency across the range of pressure ratios presented to it, and this assumption is believed to be feasible. In fact, the research from Pierobon et al. [28] suggests that it is possible to maintain a relatively con- stant isentropic turbine efﬁciency under a varying pressure ratio by modifying the speed of the turbine. In addition, at high speeds, the isentropic efﬁciency of the turbine is relatively insensitive to excursions from its operating point. Fig. 4. The comparison between the conventional ORC and the proposed dynamic ORC. or water in winter. Such undesired subcooling will increase the heat input at the evaporator and reduce the cycle efﬁciency. To avoid the undesired subcooling, the ﬂow rate of coolant passing the condenser needs to be reduced to keep the temperature of the working ﬂuid close to the designed condensation temperature. As a result, the available temperature difference (i.e., the differen- tial between the evaporation temperature and condensation tem- perature) for such a conventional ORC cycle will be kept more or less the same over the year as schematically shown by the dashed line in Fig. 4(a). Therefore, the conventional ORC has a nearly con- stant efﬁciency over the year according to Eq. (1). 2. Dynamic ORC concept In order to establish this steady-state model, it is further assumed that there is no pressure or heat loss from piping or heat exchangers, no signiﬁcant change in velocity, no change in eleva- tion, and no compressibility in the liquid ﬂow. As such, a steady- state numerical model was developed using MATLAB with REFPROP 9.1 providing the thermophysical properties of the work- ing ﬂuid [21]. An Excel ﬁle containing values of ambient temperatures for sev- eral particular locations around the world was then linked to this numerical code. From this ﬁle, the temperature of coolant available for the cycle could be obtained as the heat sink temperature for the power cycle. In contrast, a Dynamic ORC system can change the composition of its working ﬂuid in situ to match the changing ambient air tem- perature TL, and thus the available temperature difference varies over the year as shown in Fig. 4(b). The minimum available DT appears in summer, which is the same as that for the conventional ORC, while it gets much larger in winter as shown in Fig. 4(b). As such, the proposed dynamic ORC can achieve signiﬁcantly higher annual average efﬁciency than a conventional ORC according to Eq. (1). The naming convention for points in the cycle is shown in Figs. 5 and 6: The pinch point temperature difference at the condenser outlet is taken to be 5 C to facilitate the heat transfer from the working ﬂuid to the coolant (i.e., water or air), which is consistent with previous research [23,29]. An additional 2 C of sub-cooling is also added, to ensure the working ﬂuid is liquid at the pump inlet, giving the following equations: In the following sections, this paper will numerically demon- strate how the ambient temperature can be matched using this dynamic ORC concept, as well as the beneﬁts of such a Dynamic ORC power plant. T1 ¼ Tambient þ 5 ð2Þ P1 ¼ PsatðT1 þ 2Þ ð3Þ T1 ¼ Tambient þ 5 ð2Þ P1 ¼ PsatðT1 þ 2Þ ð3Þ 250 270 290 310 330 350 370 390 410 430 900 1100 1300 1500 1700 1900 Temperature (K) Entropy (kJ/kg) 1 2 3 4 2b 2c Fig. 6. Temperature–entropy diagram of a conventional ORC cycle. T1 ¼ Tambient þ 5 P1 ¼ PsatðT1 þ 2Þ ð2Þ ð3Þ P1 ¼ PsatðT1 þ 2Þ 3. Steady state numerical model 250 270 290 310 330 350 370 390 410 430 900 1100 1300 1500 1700 1900 Temperature (K) Entropy (kJ/kg) 1 2 3 4 2b 2c As shown in Fig. 5, a conventional ORC power plant has an evap- orator (boiler), an expander, a condenser, a feed pump, and a liquid storage tank. In addition to these standard components, the dynamic ORC will have a composition tuning sub system. For the convenience of demonstrating the working principle and beneﬁts of the proposed Dynamic ORC, the composition tuning subsystem can be decoupled from the main power cycle at this stage. It is assumed that the required working ﬂuid composition will be pro- vided to the cycle by the composition adjustment system to match the speciﬁed ambient temperature as required, with no other tran- sient or long-term effects. Hence, a standard steady state model can be developed to simulate the ORC with a zeotropic mixture of R134a and R245fa with a series of different compositions which Fig. 6. Temperature–entropy diagram of a conventional ORC cycle. P. Collings et al. / Applied Energy 171 (2016) 581–591 585 Four key results were calculated by the MATLAB program for each climate dataset. Firstly, the efﬁciency of the conventional ORC, gcon. This is the performance of the Organic Rankine Cycle on the hottest day of the year. Secondly, the efﬁciency of the dynamic ORC, gdyn. This is the performance of the Organic Rankine Cycle on a given day, given that day’s ambient temperature. As the steady-state model is used for these simulations, both gcon and gdyn can be calculated using Eq. (11). This allows the condenser pressure of the system to be deter- mined for the hottest day of the year, i.e. when the working ﬂuid is composed entirely of the ﬂuid component with the higher boil- ing point, in this case R245fa. The condenser pressure is assumed to remain constant throughout the change in composition of the working ﬂuid. REFPROP 9.1 allows for the calculation of each of the ﬂuid prop- erties on its extensive list given any two other properties, so tem- perature and pressure information is sufﬁcient to also calculate the enthalpy and entropy of the ﬂuid at this point. Information on the glide curves of the zeotropic mixture of R134a and R245a at this pressure as the composition changes can then be generated, as shown in Fig. 2. 3. Steady state numerical model Once this information has been obtained, the required working ﬂuid composition to satisfy the conditions of Eq. (2) (a dew point at the condenser pressure 7 C above the ambi- ent temperature) can be easily obtained for any ambient tempera- ture from the excel ﬁle. Thirdly, the annual average efﬁciency of the dynamic ORC, gdyn This is deﬁned as gdyn ¼ PN 1gdyn N ð12Þ gdyn ¼ PN 1gdyn N ð12Þ where N is the number of operation days in the year, and is assumed to be 365 here. Finally, w, the improvement in annual energy gen- eration, given by where N is the number of operation days in the year, and is assumed to be 365 here. Finally, w, the improvement in annual energy gen- eration, given by w ¼ gdyn gcon gcon 100% ð13Þ Once the required working ﬂuid composition has been estab- lished, the remainder of the cycle can be analysed using well- established principles [30,31]. A superheat of 5 C was applied to reduce the required pressure of the working ﬂuids, and the same 5 C pinch point temperature difference used in the evaporator, which allows the evaporator pressure to be calculated, using the equations ð13Þ The model was validated against experimental results obtained by Kang [32] by using the same initial parameters, and produced results that were within 2% of his values for all points of the cycle as shown in Table 1. This was considered reasonable in light of the assumptions that had been made in the production of the model. T3 ¼ Theat source 5 ð4Þ and Pevap ¼ PsatðT3 5Þ ð5Þ T3 ¼ Theat source 5 ð4Þ and Pevap ¼ PsatðT3 5Þ ð5Þ ð4Þ Table 1 i Table 1 Comparison of predictions and experimental data in the Ref. [30]. State point Present model Kang’s experiments 1 (Pump inlet) T (K) 303 303 0.00% P (bar) 1.78 1.78 0.00% h (kJ/kg) 239.1 239 0.04% s (kJ/kg K) 1.135 1.14 0.40% 2b (Saturated liquid) T (K) 350.6 350 0.16% P (bar) 7.32 7.32 0.00% h (kJ/kg) 305.4 305 0.13% s (kJ/kg K) 1.337 1.34 0.22% 2c (Saturated vapour) T (K) 350.6 350 0.16% P (bar) 7.32 7.32 0.00% h (kJ/kg) 460 460 0.00% s (kJ/kg K) 1.778 1.78 0.11% 3 (Expander inlet) T (K) 353 353 0.00% P (bar) 7.32 7.32 0.00% h (kJ/kg) 462.9 463 0.02% s (kJ/kg K) 1.786 1.79 0.22% 4 (Condenser inlet) T (K) 318.2 321 0.87% P (bar) 1.78 1.78 0.00% h (kJ/kg) 441 444 0.68% s (kJ/kg K) 1.799 1.75 2.80% gexpander ¼ ðh3 h4Þ ðh3 h4sÞ ð7Þ Once this has been done, two properties are known for each of the four key points in the cycle; pump outlet, evaporator outlet, expander outlet and condenser outlet, and so Eqs. (8)–(11) can be used to calculate the efﬁciency of the cycle. Wpump ¼ h2 h1 ð8Þ Wexpander ¼ h3 h4 ð9Þ Q evaporator ¼ h3 h2 ð10Þ gcycle ¼ Wexpander Wpump Q evaporator ð11Þ This allows for the efﬁciency of the cycle to be calculated for any ambient temperature provided by the excel spread sheet. By pop- ulating this spreadsheet with actual climate data, the year-round performance of a Dynamic Organic Rankine Cycle system can be calculated. Comparison of predictions and experimental data in the Ref. [30]. T3 ¼ Theat source 5 4. Simulation results of the dynamic ORC and and In order to verify the dynamic ORC concept as described above, a selection of case studies were carried out to demonstrate its working principle and beneﬁts. All studies were carried out using the climate data of Beijing, which has a highest temperature of around 38 C in the summer and a lowest temperature of around 17 C in the winter [20]. The monthly average temperatures are plotted in Fig. 7(a), and the error bars show the range of ﬂuctua- tion. The heat source temperature was in the range from 75 C to 200 C for these simulations. The obtained simulation results are shown in Figs. 7–9. Pevap ¼ PsatðT3 5Þ ð5Þ Pevap ¼ PsatðT3 5Þ ð5Þ To avoid the cycle becoming supercritical, which complicates analysis and comparison, the critical pressure was also calculated. A pressure of 5 kPa below this, or the pressure calculated in Eq. (5) was used as the evaporator pressure, whichever was the lower. The isentropic efﬁciency of the pump and the expander were taken to be 90% and 70%, respectively. As discussed above it is fea- sible to maintain a relatively constant isentropic turbine efﬁciency under a varying pressure ratio by modifying the speed of the tur- bine [28]. Assuming isentropic pumping and expansion, h2s and h4s can then be obtained from REFPROP, and used to calculate the actual values, using the equations: g Fig. 7(b) shows the required mass fraction of R134a in the zeo- tropic mixture to match the ambient temperature each day of the year according to Fig. 2 and Eqs. (2) and (3). On the hottest day of the summer, the ambient temperature is about 38 C, and the gpump ¼ ðh2s h1Þ ðh2 h1Þ ð6Þ gexpander ¼ ðh3 h4Þ ðh3 h4sÞ ð7Þ gpump ¼ ðh2s h1Þ ðh2 h1Þ ð6Þ Table 1 Comparison of predictions and experimental data in the Ref. [30]. Table 1 Comparison of predictions and experimental data in the Ref. [30]. Table 1 i / Applied Energy 171 (2016) 581–591 -20 -10 0 10 20 30 40 50 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Temperature (°C) (a) 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% Percentage of R134a (b) 0% 2% 4% 6% 8% 10% 12% 14% 16% 18% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Eﬃciency Dynamic ORC Convenonal ORC Case 2: TH=75°C Case 1: TH=150°C (c) en the conventional and Dynamic ORCs, for two different heat source temperatures. (a) Ambient air temperature e zeotrope (R134a and R245fa) to match the changing ambient air temperature; (c) the calculated thermal efﬁci -20 -10 0 10 20 30 40 50 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Temperature (°C) (a) (a) -20 -10 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% Percentage of R134a (b) 0% 2% 4% 6% 8% 10% 12% 14% 16% 18% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Eﬃciency Dynamic ORC Convenonal ORC Case 2: TH=75°C Case 1: TH=150°C (c) Fig. 7. Comparison between the conventional and Dynamic ORCs, for two different heat source temperatures. (a) Ambient air temperature of Beijing over a year; (b) the required composition of the zeotrope (R134a and R245fa) to match the changing ambient air temperature; (c) the calculated thermal efﬁciencies for both cycles. Fig. 7. Comparison between the conventional and Dynamic ORCs, for two different heat source temperatures. (a) Ambient air temperature of Beijing over a year; (b) the required composition of the zeotrope (R134a and R245fa) to match the changing ambient air temperature; (c) the calculated thermal efﬁciencies for both cycles. Fig. 9 shows the effect of the annual temperature variation on the improvement the dynamic cycle can achieve in annual energy production, based on simulation results holding the heat source temperature constant at 100 C. A clear increase in the beneﬁt of the dynamic cycle can be seen with increasing annual temperature variation. required mass fraction of R134a is about 10% to match this ambient condition. In the coldest day in January, the ambient temperature is around 17 C. The required mass fraction of R134a increases to around 90%. Fig. Table 1 i State point Present model Kang’s experiments 1 (Pump inlet) T (K) 303 303 0.00% P (bar) 1.78 1.78 0.00% h (kJ/kg) 239.1 239 0.04% s (kJ/kg K) 1.135 1.14 0.40% 2b (Saturated liquid) T (K) 350.6 350 0.16% P (bar) 7.32 7.32 0.00% h (kJ/kg) 305.4 305 0.13% s (kJ/kg K) 1.337 1.34 0.22% 2c (Saturated vapour) T (K) 350.6 350 0.16% P (bar) 7.32 7.32 0.00% h (kJ/kg) 460 460 0.00% s (kJ/kg K) 1.778 1.78 0.11% 3 (Expander inlet) T (K) 353 353 0.00% P (bar) 7.32 7.32 0.00% h (kJ/kg) 462.9 463 0.02% s (kJ/kg K) 1.786 1.79 0.22% 4 (Condenser inlet) T (K) 318.2 321 0.87% P (bar) 1.78 1.78 0.00% h (kJ/kg) 441 444 0.68% s (kJ/kg K) 1.799 1.75 2.80% gexpander ¼ ðh3 h4Þ ðh3 h4sÞ ð7Þ Once this has been done, two properties are known for each of the four key points in the cycle; pump outlet, evaporator outlet, expander outlet and condenser outlet, and so Eqs. (8)–(11) can be used to calculate the efﬁciency of the cycle. Wpump ¼ h2 h1 ð8Þ Wexpander ¼ h3 h4 ð9Þ Q evaporator ¼ h3 h2 ð10Þ gcycle ¼ Wexpander Wpump Q evaporator ð11Þ Wpump ¼ h2 h1 Wexpander ¼ h3 h4 Q evaporator ¼ h3 h2 ð8Þ ð9Þ ð10Þ ð11Þ This allows for the efﬁciency of the cycle to be calculated for any ambient temperature provided by the excel spread sheet. By pop- ulating this spreadsheet with actual climate data, the year-round performance of a Dynamic Organic Rankine Cycle system can be calculated. P. Collings et al. / Applied Energy 171 (2016) 581–591 586 -20 -10 0 10 20 30 40 50 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Temperature (°C) (a) 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% Percentage of R134a (b) 0% 2% 4% 6% 8% 10% 12% 14% 16% 18% Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Eﬃciency Dynamic ORC Convenonal ORC Case 2: TH=75°C Case 1: TH=150°C (c) Comparison between the conventional and Dynamic ORCs, for two different heat source temperatures. (a) Ambient air temperature of ed composition of the zeotrope (R134a and R245fa) to match the changing ambient air temperature; (c) the calculated thermal efﬁcienc P. Collings et al. Table 1 i 7(c) shows the annual variation in efﬁciency of both the conventional ORC and the dynamic ORC over the course of the year, for two different heat source temperatures, 75 and 150 C. It can be seen that in the colder months, the dynamic cycle is signiﬁcantly more efﬁcient than a conventional cycle, and also that the effect is greater when the heat source temperature is lower. 5. Composition tuning system Fig. 10 shows the basic principle behind the composition tuning system, which is essentially a distillation column. The temperature decreases up the column, and the blended working ﬂuid is injected half way up. As seen in the glide curve diagram (Fig. 2), at any given temperature, the composition of the vapour and liquid Fig. 8 shows the variation in the improvement caused by the dynamic ORC for given climate conditions (i.e., Beijing). It can be seen that the lower the heat source temperature, the greater the potential improvement, due to the low initial efﬁciency of the sys- tem making any gains proportionally more signiﬁcant. P. Collings et al. / Applied Energy 171 (2016) 581–591 587 0% 5% 10% 15% 20% 25% 30% 35% 75 95 115 135 155 175 195 ψ TH (°C) Fig. 8. The effect of heat source temperature on the value of the efﬁciency improvement w. Ambient temperatures for Beijing are used for this simulation. Fig. 8. The effect of heat source temperature on the value of the efﬁciency improvement w. Ambient temperatures for Beijing are used for this simulation. 0 5 10 15 20 25 0 10 20 30 40 50 ψ (%) Annual Temperature Variaon (℃) Fig. 9. Effect of annual temperature variation on the efﬁciency improvement w for a heat source temperature of 100 C. liquid R245fa at the bottom of the column, and nearly pure R134a vapour at the top. 0 5 10 15 20 25 0 10 20 30 40 50 ψ (%) Annual Temperature Variaon (℃) The number of theoretical trays needed for distillation can be calculated from Fenske’s Equation [33] Nmin ¼ log xD 1xD 1xB xB h i logaave ð14Þ ð14Þ where xD is the proportion of the more volatile component in the distillate, xB is the proportion of the more volatile component in the bottoms, and a is the relative volatility of the feed. This result can, as a rule of thumb, be doubled to give the actual number of trays required [33]. The number of trays against the purity of product the resulting distillation column will produce is plotted below in Fig. 11, show- ing the increase in purity of the product caused by increasing the number of trays in the distillation column. Its implementation in our Dynamic ORC power plant would be as shown in Fig. 12. 5. Composition tuning system Power required for distillation and compression against the time to perform it. Fig. 12. Schematic diagram of the composition tuning sub-system. where _F is the feed ﬂow rate, r is the heat of vaporisation of the feed, xFa is the fraction of lighter component in the feed, and a is the rel- ative volatility, or the ratio of vapour pressures of the two compo- nents. This theoretical value of Qmin is estimated by Stichlmair and Stemmer to be about 90% that of a practical system. This will give the energy required for complete separation of a given composition of feed. The energy required to replace the entire charge of an ORC system can be obtained by dividing the process into 100 steps, and summing them as Qtotal ¼ X 100 i¼0 Q i ð17Þ Qtotal ¼ X 100 i¼0 Q i ð17Þ In this way, the total energy required to distil the entire charge of the system was calculated using MATLAB to be 613 MJ, giving an average annual distillation power of 19.42 W. In order to compress 1 kg of R134a vapour from condenser pres- sure (2.9 bar) to storage pressure (15 bar to ensure liquid condition at 40 C) requires 30 kJ of energy [20]. Over the course of a year, the entire charge of the system needs to be removed and compressed to storage pressure, so the total energy requirement for the com- pressor is 120 MJ. Averaged over the course of the year, this leads to a parasitic compressor power of 3.8 W. This results in a total average energy draw for the distillation system of 23.22 W year- round. Fig. 12. Schematic diagram of the composition tuning sub-system Refrigeration and heat pump systems generally have a refriger- ant charge of 0.25–0.5 kg/kW installed capacity [34]. This power ﬁgure refers to the amount of heat the heat pump can transfer, which corresponds to the evaporator and condenser loading in an ORC system. As per the simulations used in our case study, the efﬁciency for an ORC with a heat source temperature of 100 C operating under Beijing’s ambient conditions has an efﬁ- ciency of 9.89% meaning that the power transferred through the evaporator and condenser is taken to be between 10.11 and 9.11 times the energy extracted through the expander, respectively. This implies that Organic Rankine Cycles will have a system charge of 2.278–5.05 kg/kW. 5. Composition tuning system This will give the energy required for complete separation of a given composition of feed. The energy required to replace the entire charge of an ORC system can be obtained by dividing the process into 100 steps, and summing them as Qtotal ¼ X 100 i¼0 Q i ð17Þ In this way, the total energy required to distil the entire charge of the system was calculated using MATLAB to be 613 MJ, giving an average annual distillation power of 19.42 W. In order to compress 1 kg of R134a vapour from condenser pres- sure (2.9 bar) to storage pressure (15 bar to ensure liquid condition at 40 C) requires 30 kJ of energy [20]. Over the course of a year, the i h f h d b d d d 90 91 92 93 94 95 96 97 98 99 100 0 5 10 15 20 Purity of product (%) Number of trays Fig. 11. Effect of number of trays on product purity. Fig. 12. Schematic diagram of the composition tuning sub-system. 0 2 4 6 8 10 12 14 0 10 20 30 40 50 60 70 80 90 100 Disllaon power (kW) Response Time (hours) Qevap Wcomp Wcomp Qevap Fig. 13. Power required for distillation and compression against the time to perform it. 90 91 92 93 94 95 96 97 98 99 100 0 5 10 15 20 Purity of product (%) Number of trays Fig. 11. Effect of number of trays on product purity. 0 2 4 6 8 10 12 14 0 10 20 30 40 50 60 70 80 90 100 Disllaon power (kW) Response Time (hours) Qevap Wcomp Wcomp Qevap Fig. 13. Power required for distillation and compression against the time to perform it. 0 2 4 6 8 10 12 14 0 10 20 30 40 50 60 70 80 90 100 Disllaon power (kW) Response Time (hours) Qevap Wcomp Wcomp Qevap Fig. 13. Power required for distillation and compression against the time to perform it. 0 2 4 6 8 10 12 14 0 10 20 30 40 50 60 70 80 90 100 Disllaon power (kW) Response Time (hours) Qevap Wcomp Wcomp Qevap 90 91 92 93 94 95 96 97 98 99 100 0 5 10 15 20 Purity of product (%) Number of trays Fig. 11. Effect of number of trays on product purity. Fig. 13. 5. Composition tuning system However, in the event of extreme weather conditions, such as cold snaps or heat waves, it is desirable to have the capacity to completely replace the working ﬂuid in a short period of time, e.g., 12 h for the worst case scenario. This results in a compressor power of 2.8 kW, and a distillation column heat demand of 14.26 kW, according to the equations: _Wcomp;installed ¼ 365 24 12 _Wcomp;average ð18Þ _Qcolumn;installed ¼ 365 24 12 _Qcolumn;average ð19Þ ð18Þ The charge of working ﬂuid for a 1 MWe plant would therefore be estimated in the range of 2778–5050 kg. This paper assumes a system charge of 4000 kg. The two greatest sources of energy required for the composition tuning system are the energy required to vaporise the working ﬂuid in the distillation column, and the power required to compress the distilled R134a to storage pressure. ð19Þ Fig. 13 shows the required power for the distillation column and the compressor with a varying response time of the system. It can be seen that the shorter the response time, the larger the required capacity of the distillation system. This data is used in the following section to perform an economic analysis of the system. Therefore, the amount of energy required to switch out the working ﬂuid can be given by Eq. (15) Ecomposition tuning ¼ Ecolumn þ Ecompressor ð15Þ ð15Þ The heat required for complete separation of a binary mixture is given by Stichlmair and Stemmer [35] to be 5. Composition tuning system The liquid R245fa collects at the bottom of the distillation column, where it can be pumped to a storage tank. The system is designed such that R245fa can be stored as a liquid at condenser pressure, so no extra pressurization is needed. The R134a vapour collects at the top of the system. A compressor then increases the pressure of the vapour to a level at which it can also be stored as a liquid at the ambient temperature. This causes an increase in the temperature of the vapour, so it must be passed through a post-cooler before collecting in the storage tank. The working ﬂuid removed from the power generation cycle can be replaced by simply opening a valve and pumping the desired component back into the system. Fig. 9. Effect of annual temperature variation on the efﬁciency improvement w for a heat source temperature of 100 C. phases can be obtained. The design of the column is such that the vapour ﬂows upwards due to buoyancy, whereas the liquid rains towards the bottom of the column due to gravity. Each tray in the distillation column exists at a different temperature, with the cooler trays located towards the top, allowing liquid richer in R245fa to rain out even as its concentration in the overall ﬂuid decreases. Eventually, the vapour at the top of the column has been almost completely depleted of R245fa, and the liquid at the bottom has had almost all of the R134a driven out of it, leaving nearly pure R134a out R245fa out Fluid Mixture In Down flowing Liquid Up flowing Vapour Lowest Temperature Highest Temperature Liquid composition Vapour composition Fig. 10. Distillation process for separating the components of the zeotropic ﬂuid used in this research. Lowest Temperature Liquid composition Vapour composition Liquid composition Vapour composition Down flowing Liquid Fluid Mixture In R245fa out Highest Temperature Fig. 10. Distillation process for separating the components of the zeotropic ﬂuid used in this research. P. Collings et al. / Applied Energy 171 (2016) 581–591 588 where _F is the feed ﬂow rate, r is the heat of vaporisation of the feed, xFa is the fraction of lighter component in the feed, and a is the rel- ative volatility, or the ratio of vapour pressures of the two compo- nents. This theoretical value of Qmin is estimated by Stichlmair and Stemmer to be about 90% that of a practical system. Table 2 C fﬁi log Zp ¼ K1 þ ðK2 log XÞ þ ðK3ðlog XÞ 2Þ ð20Þ ð20Þ where Zp is the component price in US Dollars, K1, K2 and K3 are con- stants as deﬁned in Table 2, and B is a characteristic of the compo- nent under analysis, this being power for ﬂuid machinery and surface area for heat exchangers, which was taken to be 2 m2/kW, as per Zhang et al. [36]. where Zp is the component price in US Dollars, K1, K2 and K3 are con- stants as deﬁned in Table 2, and B is a characteristic of the compo- nent under analysis, this being power for ﬂuid machinery and surface area for heat exchangers, which was taken to be 2 m2/kW, as per Zhang et al. [36]. The generator cost is calculated using the following equation [31] Zp ¼ 1; 850; 000Wexpander 11; 8000:94 ð21Þ ð21Þ In order to successfully operate a dynamic cycle such as the one described in this paper, the amount of working ﬂuid in the system would need to be doubled compared to a conventional ORC of the same installed capacity. On the hottest day of the year, the working ﬂuid in the power cycle is 100% R245fa. An equal mass of R134a is required in storage to completely replace this for the coldest day of the year. As discussed above, this would increase the installation cost of the system by £100,000, or 6.01%. In theory, this could be reduced by using component ﬂuids with a greater difference between their boiling points, and therefore avoiding the need to use the full range of compositions to match the variation in temperature. These component prices assume that they are made from car- bon steel, and operate at ambient pressure. If they are made of other materials, or operate at other pressures, a correction factor must be applied, according to the following equations. ZBM ¼ ZPFBM ð22Þ FBM ¼ B1 þ ðB2FMFPÞ ð23Þ ð22Þ ZBM ¼ ZPFBM FBM ¼ B1 þ ðB2FMFPÞ ð23Þ where ZBM is the corrected cost, FBM is an overall correction factor, B1 and B2 are correction factors given in, and FM and FP are correc- tion factors for material and operating pressure respectively. Table 2 C fﬁi the composition tuning system. As the distillation system consists of heat exchangers, pumps and turbo-machinery, an assumption was made that the component costs were of the same order as that of an ORC system of comparable size. This allowed the system data assembled by Quoilin et al. [37] to be used estimate a cost for the composition tuning system. the composition tuning system. As the distillation system consists of heat exchangers, pumps and turbo-machinery, an assumption was made that the component costs were of the same order as that of an ORC system of comparable size. This allowed the system data assembled by Quoilin et al. [37] to be used estimate a cost for the composition tuning system. in Beijing, and considering the sizing of the critical components, the extra working ﬂuid, and the extra energy consumption to run the composition tuning system. This was used to calculate the Pay- back Period and Net Present Value of a case study of a system util- ising a 100 C heat source. Section 5 calculated the worst case scenario of having to replace the entire charge of the system in 12 h, giving a required compres- sor capacity of 2.1 kW, and a heat requirement for the distillation column of 14.6 kW. The cost of each major component was calculated using the Chemical Engineering Component Price Index, as utilised by Wang et al. [31], and outlined in the following equations Quoilin et al.’s data shows that ORC systems in this power range have a cost of €2000–8000/kWe, which is equivalent to £1500– 6000/kWe. If we assume an average thermal efﬁciency of 10%, this translates to a cost of £150–600/kW thermal energy. We can there- fore roughly estimate the cost of the composition tuning system as £150–600/kW thermal energy. For the worst case scenario, the cost of the composition tuning system is estimated as £1600–6400, but this cost drops off sharply with an increasing response time of the system, as shown in Fig. 14, stabilising at a maximum of about £800 for longer timescales, such as would be experienced if the composition tuning system were operated for a short period each day to keep pace with seasonal temperature shifts. 6. Economic analysis _Q min ¼ _Fr 1 a 1 þ xFa ð16Þ _Q min ¼ _Fr 1 a 1 þ xFa An economic analysis of the system was carried out, focusing on the case study of a 1 MW power plant with the climate conditions ð16Þ P. Collings et al. / Applied Energy 171 (2016) 581–591 589 Table 3 Component prices for the conventional ORC power plant. Component Cost % Pump £298,915 17.96 Evaporator £84,056 5.05 Expander £441,075 26.49 Condenser £468,302 28.13 Working ﬂuid £100,000 6.01 Generator £272,419 16.36 Total £1,664,767 100 Table 2 Coefﬁcients for component pricing [31]. Pump Evaporator Expander Condenser Generator K1 3.3892 4.3147 2.2476 4.0336 K2 0.0536 0.3030 1.4965 0.2341 K3 0.1538 0.1634 0.1618 0.0497 X 40 kW 2000 m2 1000 kW 2000 m2 1000 kW C1 0.3935 0.03881 C2 0.3957 0.11272 C3 0.00226 0.08183 B1 1.89 1.63 3.5 1.96 B2 1.35 1.66 1.21 Fm 1.5 1 1 FBM 3.5 1.5 Table 2 C fﬁi Where a value of FP is not directly given, it can be calculated using the equation [31] Table 4 shows that the comparison of the costs between a dynamic ORC system and a conventional ORC power plant. The log Fp ¼ C1 þ ðC2 log PÞ þ ðC3ðlog PÞ 2Þ ð24Þ ð24Þ 0 1 2 3 4 5 6 7 8 12 17 22 27 32 37 42 47 52 57 62 67 72 77 82 87 92 97 Tuning system Cost (£1000) Response Time (hours) Minimum (£150/kWth) Average (£375/kWth) Maximum (£600/kWth) Fig. 14. Tuning system cost against desired response time. 0 1 2 3 4 5 6 7 8 12 17 22 27 32 37 42 47 52 57 62 67 72 77 82 87 92 97 Tuning system Cost (£1000) Response Time (hours) Minimum (£150/kWth) Average (£375/kWth) Maximum (£600/kWth) where P is the operating pressure of the component. where P is the operating pressure of the component. where P is the operating pressure of the component. Current market prices of R245a are roughly £25/kg at the time of writing this paper, giving a cost of £100,000 for the 4000 kg required for a 1 MW system. Using Eqs. (20)–(24), and an exchange rate of £0.66 to the US dollar, the cost of the components for a conventional 1 MW ORC system is estimated as follows in Table 3: Quoilin et al. [37] have collated data on real-world ORC Sys- tems, plotting the price per kW against the system size, for fully- commissioned systems, and for the module costs only. Their ﬁnd- ings show that the module-only cost for systems in the 1 MW range is €1000–3000 per kW (equivalent to £750–2250). The ﬁgure generated by the above analysis falls right in the middle of this range, which suggests it is a reasonable estimation. 12 17 22 27 32 37 42 47 52 57 62 67 72 77 82 87 92 97 Response Time (hours) 7 42 47 52 57 62 67 72 Response Time (hours) The predictions of this method do not hold for smaller compo- nents, so an approximation had to be made to estimate the cost of Fig. 14. Tuning system cost against desired response time. Fig. 14. Tuning system cost against desired response time. P. Collings et al. 7. Conclusions This paper has proposed a novel Dynamic ORC concept, which uses a binary zeotropic mixture as the working ﬂuid and can dynamically alter mixture composition to match the ambient air temperature, so that the resultant power plant can achieve higher annual average efﬁciency than a conventional ORC using a single component working ﬂuid. A numerical model based on the ther- mophysical properties provided by REFPROP 9.1 has been devel- oped to simulate the proposed Dynamic ORC. The simulation results have demonstrated that a dynamic ORC can improve annual energy generation in a continental climate such as Beijing by over 20% when the heat source temperature is of the order of 100 C. Furthermore, as the heat source temperature decreases, the bene- ﬁts of Dynamic ORC become more signiﬁcant, showing that the Dynamic ORC has great potential for low temperature applications. An economic analysis was then carried out for the case study of a dynamic ORC power plant with 1 MW electricity output under Beijing’s climate conditions. The costs of the power plant together with the composition tuning system were estimated according to published data and standard methods in literature. Compared to a conventional ORC power plant, the extra costs fall into two cate- gories: the ﬁrst being the extra volume of working ﬂuid, and the second being the composition tuning system, essentially a small distillation system. The cost of the extra working ﬂuid is estimated to be in the range of 6%. However the costs of the composition tun- ing system strongly depend the operating time required to com- plete the separation of the two components. For the worst case scenario, the whole charge needs to be separated in several hours. The required composition system in this case will add about 0.4% to the total cost of the power plant. This value starts to become negligible as the response time increases to over 100 h. where N is the number of years into the future, RN is the annual rev- enue from the system, d is the discount rate, and C is the capital employed, or the total initial investment in the ORC system. This metric is considered by Lecompte et al. to be the best for compar- ison of similar systems, although it does make assumptions that are dependent on time and location [40]. 7. Conclusions Systems can also be com- pared on the basis of Return on Capital Employed This paper has proposed a novel Dynamic ORC concept, which uses a binary zeotropic mixture as the working ﬂuid and can dynamically alter mixture composition to match the ambient air temperature, so that the resultant power plant can achieve higher annual average efﬁciency than a conventional ORC using a single component working ﬂuid. A numerical model based on the ther- mophysical properties provided by REFPROP 9.1 has been devel- oped to simulate the proposed Dynamic ORC. The simulation results have demonstrated that a dynamic ORC can improve annual energy generation in a continental climate such as Beijing by over 20% when the heat source temperature is of the order of 100 C. Furthermore, as the heat source temperature decreases, the bene- ﬁts of Dynamic ORC become more signiﬁcant, showing that the Dynamic ORC has great potential for low temperature applications. ROCE ¼ PN 0 RN C ROCE ¼ PN 0 RN C ð26Þ ð26Þ where Capital Employed is the initial investment in the ORC system. where Capital Employed is the initial investment in the ORC system. Figure 15 shows the net present value of both the dynamic ORC and conventional ORC power plants. The heat source temperature is given as 100 C, while different climate conditions (i.e., the annual temperature variations) have been used to achieve different values of efﬁciency improvement w. Moreover, for the convenience of comparison, the summer temperature is kept as the same for all cases, while the winter temperature varies. As such, the effect of the value of w on the power plants’ payback period can be com- pared. The comparison clearly shows that the dynamic ORC power plant has the higher NPV. Furthermore, the higher the value of efﬁ- ciency improvement w, the higher the NPV. An economic analysis was then carried out for the case study of a dynamic ORC power plant with 1 MW electricity output under Beijing’s climate conditions. The costs of the power plant together with the composition tuning system were estimated according to published data and standard methods in literature. Compared to a conventional ORC power plant, the extra costs fall into two cate- gories: the ﬁrst being the extra volume of working ﬂuid, and the second being the composition tuning system, essentially a small distillation system. Table 5 The price of electricity was taken to remain constant at £0.05/ kW h in the UK. The discount rate, which reﬂects the current value of future energy generation, was taken to be 10%, which is in agree- ment with other small- to medium-scale energy generation pro- jects [38,39]. This allowed the Net Present Value of the conventional and dynamic concepts to be compared over time, according to the equation period is around 19.3% shorter than for the conventional cycle. The dynamic cycle has a higher NPV than the conventional cycle for all plant lifetimes greater than 3 years. period is around 19.3% shorter than for the conventional cycle. The dynamic cycle has a higher NPV than the conventional cycle for all plant lifetimes greater than 3 years. NPV ¼ C þ X Years N¼0 RN ð1 þ dÞ N ð25Þ ð25Þ Table 4 Table 4 Economic analysis of the dynamic ORC power plant. Table 4 Economic analysis of the dynamic ORC power plant. composition tuning system will introduce around 0.39% of costs, while the extra working ﬂuid will introduce about 6.01% of the costs. Overall, the total costs of a dynamic ORC power plant is pro- jected to be 106.4% that of a conventional system of the same capacity. This increased cost will be offset by a greater energy production. p Quoilin et al. [37] have collated the full-plant prices of several existing ORC plants which vary from €2000 to €4000/kWe (equiv- alent to £1500 to £3000/kWe) installed capacity for systems with a capacity over 1 MWe. For the following economic calculations this paper will use an intermediate value of £2500/kWe for our case study using a 100 C heat source, which accounts for additional installation and commissioning costs over and above the module costs calculated earlier and presented in Table 4. Higher tempera- ture heat sources will tend to lead to greater efﬁciency, and reduce the cost per kW of the system due to the smaller relative size of components, especially the heat exchangers. Fig. 15. NPV over time for the conventional cycle and three different values of w (with a capital cost of £2500/kW). Table 5 Comparison of economic data for differing values of w (£2500/kW). Payback period (months) NPV (20 years) ROCE Conventional cycle 114 £963,000 1.39 Dynamic ORC: w = 10% 106 £1,189,000 1.45 Dynamic ORC: w = 15% 97 £1,362,000 1.52 Dynamic ORC: w = 20% 92 £1,545,000 1.59 period is around 19.3% shorter than for the conventional cycle. The dynamic cycle has a higher NPV than the conventional cycle for all plant lifetimes greater than 3 years. Table 5 Comparison of economic data for differing values of w (£2500/kW). Payback period (months) NPV (20 years) ROCE Conventional cycle 114 £963,000 1.39 Dynamic ORC: w = 10% 106 £1,189,000 1.45 Dynamic ORC: w = 15% 97 £1,362,000 1.52 Dynamic ORC: w = 20% 92 £1,545,000 1.59 Table 5 Comparison of economic data for differing values of w (£2500/kW). Table 2 C fﬁi / Applied Energy 171 (2016) 581–591 590 -3000 -2500 -2000 -1500 -1000 -500 500 1000 1500 2000 0 5 10 15 20 Net Present Value (1000£) Years Convenonal ψ=10% ψ=15% ψ=20% Fig. 15. NPV over time for the conventional cycle and three different values of w (with a capital cost of £2500/kW). Table 4 Economic analysis of the dynamic ORC power plant. Conventional ORC Composition tuning system Extra working ﬂuid Dynamic ORC Cost £1,639,767 £6400 £100,000 £1,746,167 % 100 +0.39 +6.01 106.4 7. Conclusions The cost of the extra working ﬂuid is estimated to be in the range of 6%. However the costs of the composition tun- ing system strongly depend the operating time required to com- plete the separation of the two components. For the worst case scenario, the whole charge needs to be separated in several hours. The required composition system in this case will add about 0.4% to the total cost of the power plant. This value starts to become negligible as the response time increases to over 100 h. Extracting the data from Fig. 15, Table 5 shows the change in NPV for varying values of efﬁciency improvement w in detail. 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W4389532419.txt	https://aclanthology.org/2023.emnlp-main.824.pdf	en		ViStruct: Visual Structural Knowledge Extraction via Curriculum Guided Code-Vision Representation	null	2,023	cc-by	9,763	ViStruct: Visual Structural Knowledge Extraction via Curriculum Guided Code-Vision Representation Yangyi Chen1 , Xingyao Wang1 , Manling Li2 , Derek Hoiem1 , Heng Ji1 1 University of Illinois Urbana-Champaign 2 Northwestern University {yangyic3, xingyao6, manling2, dhoiem, hengji}@illinois.edu Abstract State-of-the-art vision-language models (VLMs) still have limited performance in structural knowledge extraction, such as relations between objects. In this work, we present ViStruct, a training framework to learn VLMs for effective visual structural knowledge extraction. Two novel designs are incorporated. First, we propose to leverage the inherent structure of programming language to depict visual structural information. This approach enables explicit and consistent representation of visual structural information of multiple granularities, such as concepts, relations, and events, in a well-organized structured format. Second, we introduce curriculum-based learning for VLMs to progressively comprehend visual structures, from fundamental visual concepts to intricate event structures. Our intuition is that lower-level knowledge may contribute to complex visual structure understanding. Furthermore, we compile and release a collection of datasets tailored for visual structural knowledge extraction. We adopt a weakly-supervised approach to directly generate visual event structures from captions for ViStruct training, capitalizing on abundant image-caption pairs from the web. In experiments, we evaluate ViStruct on visual structure prediction tasks, demonstrating its effectiveness in improving the understanding of visual structures. The code is public at https://github.com/ Yangyi-Chen/vi-struct. 1 Figure 1: Programming language enables a unified representation of visual structural information in codevision representations, including concepts, attributes, locations, relations, and visual events. Introduction Vision-language models (VLMs) exhibit impressive performance across various multimodal downstream tasks (Li et al., 2019a; Tan and Bansal, 2019; Chen et al., 2023b; Wang et al., 2022a), such as visual question answering (Antol et al., 2015; Yuan et al., 2023), image captioning (Karpathy and FeiFei, 2015), visual event extraction (Li et al., 2022b; Wang et al., 2022c), chart understanding (Zhou et al., 2023), and multimedia misinformation detection (Fung et al., 2021; Luo et al., 2021). The recipe lies in the pre-training on web-scale data, enabling substantial enhancements in vision-language representation (Radford et al., 2021; Li et al., 2022a). However, recent evaluations expose the limitations of existing VLMs models in capturing visual structural information (Hendricks and Nematzadeh, 2021; Zhao et al., 2022; Yüksekgönül et al., 2022), rendering them ineffective for visual structural knowledge extraction. For example, Yüksekgönül et al. (2022) argue that VLMs represent images as bag-of-words, disregarding relations among objects. In this work, we present ViStruct, a training framework for VLMs to improve their visual structural knowledge extraction ability. There are two key challenges in multi-granularity visual structural knowledge learning: (1) capturing the structures at each granularity, for which we propose code-vision representations, and (2) recognizing the dependencies between different granularities, for which we propose a curriculum pyramid to guide the knowledge learning. The motivation of employing code-vision rep- 13342 Proceedings of the 2023 Conference on Empirical Methods in Natural Language Processing, pages 13342–13357 December 6-10, 2023 ©2023 Association for Computational Linguistics resentations is to mitigate the negative impact of commonly employed techniques such as contrastive learning (Radford et al., 2021) and imageconditioned language modeling (Li et al., 2023), which limit VLMs to focus solely on primary objects, resulting in “bag-of-words” representations. we draw inspiration from prior successful application of programming language in representing linguistic structures (Madaan et al., 2022; Wang et al., 2023b). We propose to extend this “code for structure representation” principle to multimodal settings for explicit representation of visual structures. Initial attempts to employ code representation in visual reasoning (Gupta and Kembhavi, 2023; Surís et al., 2023) focus solely on object information and lack perception abilities, but relying on the off-the-shelf object detectors as tools. As a result, we establish a multi-granularity code representation framework to capture structured visual information, including visual concepts, attributes, relations, and events (see Figure 1). To further capture the interdependencies among various granularities, we propose the curriculum guided visual structural knowledge learning. Our intuition is that foundational knowledge (such as concepts) may enhance comprehension of more complex visual structures (such as visual events). Thus, unlike previous work that involves pretraining VLMs on a mixture of tasks, we build a visual structural knowledge pyramid (see Figure 2), aligning with the pedagogical strategy of progressively addressing simpler to more complex tasks during training (Soviany et al., 2022; Wang et al., 2022b). However, in contrast to conventional curriculum learning methods that focus on arranging data samples to facilitate training for a specific task (Jiang et al., 2014a; Spitkovsky et al., 2010b), our objective is to enable VLMs to understand visual structures comprehensively at all levels. To accomplish this, we incorporate a replay buffer mechanism that selectively reserves a small amount of data at each level to preserve lower-level knowledge throughout the training process. Moreover, we curate an extensive collection of datasets for ViStruct, called ViStruct Suite. This suite includes multi-level structural knowledge, aligned with the curriculum pyramid. We first collect existing labeled datasets and large-scale unlabeled image-caption pairs. Then, we adopt a weakly-supervised approach to facilitate the creation of visual event structure labels from captions in a scalable manner. To ensure consistent class label definitions, we automate the alignment of all concepts from various datasets to WordNet (Miller, 1995) and FrameNet (Baker et al., 1998) synsets, with minimal manual assistance. For downstream task evaluation, we find that ViStruct consistently outperforms various baselines on three downstream structure prediction tasks (visual relation detection, scene graph classification, and situation recognition). In addition, we find that code representation and curriculum learning both provide consistent gains on downstream tasks and demonstrate their effectiveness on zero-shot visual structure prediction. Parsing visual data into various levels of detail has been a longstanding problem. To the best of our knowledge, we are the first to extract multigranularity visual structured knowledge via codevision representation learning, which effectively captures the hierarchy among different levels of visual knowledge, offering a comprehensive and compositional understanding of overall structure. 2 2.1 Related Work Vision-Language Models (VLMs) for Visual Knowledge Extraction Visual knowledge extraction has been long optimized as separate tasks for each granularity of extraction, such as object detection, relation detection, scene graph parsing, situation recognition and event extraction, as further detailed in Appendix B. Recent years witness great success through VLMs to obtain knowledge of visual concepts (Sadhu et al., 2021) and events (Wen et al., 2021; Deshpande et al., 2022; Li et al., 2022c). The primary goal of VLMs is learning representations from multiple modalities to capture visual details and their alignment with text semantics (Gan et al., 2022; Uppal et al., 2022; Wang et al., 2022c), and the corresponding research can be broadly classified into three stages. The initial stage of VLMs primarily relies on off-the-shelf object detectors to represent visual modality (Li et al., 2019b; Tan and Bansal, 2019; Lu et al., 2019; Li et al., 2020a) and learn the alignment across modalities (Li et al., 2020c, 2021b). The second phase of VLMs aims to eliminate the use of resource-intensive object detectors and address the issue of unseen objects (Dou et al., 2022; Huang et al., 2020; Kim et al., 2021; Huang et al., 2021; Xu et al., 2021; Yang et al., 2022; Yao et al., 13343 Figure 2: The curriculum pyramid in ViStruct that incorporates five different levels of multimodal knowledge acquisition, progressing from basic to more advanced stages. All levels of visual structures can be uniformly represented using the programming language. The code is generated based on the image in Figure 1. 2022). Large-scale multimodal data is required and emphasized to better align two modalities (Li et al., 2021a; Radford et al., 2021; Jia et al., 2021). The current stage of VLMs moves towards a more unified pre-training goal to perform various tasks without task-specific adaptations (Wang et al., 2021, 2022a; Li et al., 2023). Some research leverages programming language (Gupta and Kembhavi, 2023; Surís et al., 2023), but lacks perception ability and relies on pre-trained object detectors. Although we witness great success of existing VLMs on benchmark evaluations, in-depth diagnostic investigations have exposed their shortcomings in fundamental visual concept identification (Hendricks and Nematzadeh, 2021; Zhao et al., 2022; Yüksekgönül et al., 2022), raising doubts about whether current VLMs are overemphasizing objects to represent images while ignoring other visual structures, akin to the initial stage VLMs, making them inadequate in structural knowledge extraction. Thus, in this work, we design a training framework to improve VLMs’ visual structural knowledge extraction. 2.2 Curriculum Learning Curriculum learning has proven to be a useful technique for improving the learning process by gradually exposing the learner to more challenging examples (Bengio et al., 2009; Karras et al., 2018; Wang et al., 2020; Su et al., 2021; Platanios et al., 2019; Yu et al., 2022). However, it has been studied mainly in a more limited form, ordering and presenting examples to train a single task (Bengio et al., 2009; Spitkovsky et al., 2010a; Kumar et al., 2010; Jiang et al., 2014b; Graves et al., 2017). In this work, we propose the first task-level, multifaceted, multimodal approach to curriculum learning for visual structural knowledge learning. 3 ViStruct We propose ViStruct that learns a vision-language model that takes images as input and generates a set of structural knowledge, such as concepts, relations, and events involved in the images (see Figure 2). We first define a code-vision representation (Sec. 3.1), which serves as the output to decode visual structural knowledge as code blocks. We then propose a curriculum pyramid to guide the training objectives from low-level towards high-level semantics (Sec. 3.2). To train the model, we construct a comprehensive multi-granularity dataset ViStruct Suite (Sec. 3.3), and optimize the model only on information that describe relavant visual strcutural information (Sec. 3.4). 3.1 Code-Vision Representation Visual structural information depicts both visual knowledge elements (such as visual concepts, locations, attributes) and complex structural connections between them (such as relations, and visual events with associated arguments). To encode visual structural knowledge, the first question is to define the appropriate structured representation. Such a representation is required to not only have robust capabilities for illustrating structures and their intricate interconnections, but also be unified and consistent across multiple levels of semantic granularity. It is also expected to be adaptable and extendable to accommodate other future semantic granularities. In order to address these requirements, we leverage the inherent capabilities of programming language (PL). We adopt Python as the PL in this paper due to its straightforwardness and popularity, allowing us to effectively harness its structured representation power and well-resourced data for code-vision pretraining. Similar to PL, we define ViStruct including both class definition and 13344 Visual Structural Information & Definition PL class definition Visual structure Event arguments/subject (object) in relation Human-written definition Class name Arguments for class instantiation Docstring Table 1: The alignment of visual structures, visual semantic roles, annotated definitions, and programming language (PL) class definitions. The concrete examples are shown in Figure 3. Algorithm 1 The training algorithm described in Sec. 3.4. We omit the warm-up process for brevity. 1: 2: 3: 4: 5: 6: 7: 8: Pθ ▷ a pre-trained vision-language model Dstage = [DCR , DOG , DOA , DOR , DE ] ▷ Stage-specific datasets defined in Sec. 3.2 DRB = {} ▷ Replay buffer in Sec. 3.2 for Ds in Dstage do Dtrain = Ds ∪ DRB Finetune M using Dtrain DRB = DRB ∪ random_subset_of(Ds ) end for niently represented in a unified and structured manner. In this work, we consider the most salient visual structural information, including concepts, locations, attributes, relations, and visual events with associated arguments. The structure-code mapping is detailed in Table 2. It is noteworthy that our codevision representation can be effortlessly expanded to encompass supplementary visual details, such as affordance, object states, and similar aspects. Figure 3: Examples of the class definitions of basic visual concepts and visual events. instantiation of visual structural information. Class Definition of Visual Structural Information. To define a general set of multi-granularity visual structures, we combine two external knowledge resources: WordNet (Miller, 1995) for basic visual concepts and relations; and FrameNet (Baker et al., 1998) for visual events. Class definitions in PL provide a unified and structured way to represent the visual structures, semantic roles, and human-written definitions in WordNet and FrameNet. Table 1 details the structure-class mapping we defined. For each visual concept, we define object location loc for object grounding, and attributes for object properties, as shown in the “horse” example in Figure 3. On top of that, we define visual events, which have visual concepts as participants, characterized by their semantic roles (such as agent and create_entity in the example visual event “building” in Figure 3). As discussed in Sec. 3.4, these definitions are used to warm up the model through finetuning. Instantiation of Visual Structural Information. As shown in Figure 1, the multi-granularity visual structural information of each image can be conve- 3.2 Curriculum-Based Training for Knowledge Extraction with Replay Buffer Existing vision-language pre-training approaches are mostly based on end-to-end training on a mixture of tasks (Li et al., 2023), which operates differently from the way humans learn. In human learning, for example, the well-validated Montessori method (Lillard et al., 2017) focuses on a curriculum of self-directed learning in stages, where demonstrated mastery of elementary concepts is required before advancing to those more complex. Based on this insight, we propose a curriculum pyramid that directs VLMs towards progressively learning to extract visual structural knowledge from simpler to more complex concepts. In particular, we introduce the replay buffer mechanism to prevent the forgetting of foundational structural knowledge obtained at the lower levels of the curriculum. The training algorithm is described in Alg. 1. Curriculum Pyramid. We present our curriculum pyramid in Figure 2. We categorize the process of learning complete visual structural knowledge into five levels, based on the assumption that knowledge acquisition at lower levels (such as concepts) can facilitate the acquisition of higher-level knowledge (such as visual events). For example, in Figure 1, identifying the “horse” and “grass” concepts contributes to understanding the “eat” event with 13345 Structural Information Concept Grounded Object Attribute Relation Event Python Code Representation object = <object_synset>(loc=None, attributes=None) object = <object_synset>(loc=[pos_1, pos_2, pos_3, pos_4], attributes=None) object = <object_synset>(loc=[pos_1, pos_2, pos_3, pos_4], attributes=[<attribute_synset>]) <relation_synset>(sub=<defined_object>, obj=<defined_object>) <event>(<argument_name>=<object_name>, <argument_name>=<object_name>) Table 2: The correspondence between visual structural information and Python code representations. Figure 4: We generate the visual events with argument roles from the captions using the off-the-shelf system. these two participants. Conditioned on an image input, we train the model to reconstruct masked visual structures represented using code (e.g., arguments). Specifically, we mask out targeted visual information in the code and treat this masked sequence along with the image as input to the model. For example (Figure 1), in object attributes learning stage, we mask out the target attributes: object = horse(loc=[pos_1, pos_2, pos_3, pos_4], attributes=[<mask>]). Then we train the model to generate the code sequence replacing the <mask> with “brown” and “hungry”. Formally, given an image i, the code sequence s and a mask function mask(·) to mask different information for different stages. We use an auto-regressive loss to train the code-vision representation model on the dataset D, that is, we are minimizing the following loss function: X L(θ) = − logPθ (s\|i, mask(s)) (i,s)∈D where θ refers to the model parameters, D is the dataset we used for training, which composed of a stage-specific dataset Dspecific ∈ {DCR , DOG , DOA , DOR , DE } for all the five stages in Figure 2 and a replay buffer DRB , that is, D = Dspecific ∪ DRB for each stage. We use Figure 1 as a running example to introduce each stage in the pyramid. The initial stage involves basic Concept Recognition (CR), which applies VLMs to identify the most prominent objects in the images presented (e.g., horse and grass). In a recent study by Kamath et al. (2022); Whitehead et al. (2021), it is demonstrated that decoupling the acquisition of basic con- cepts from the learning of task-specific skills is effective. We adopt this approach and anticipate that VLMs will gain a range of fundamental visual concepts, including rare object categories, at this level. At the second level, VLMs are required to perform Object Grounding (OG), which involves the identification of objects with specific, concrete locations in the images. The third level places emphasis on Object Attribute (OA) detection, with the goal of inferring the attributes associated with particular objects (e.g., the horse is brown and hungry). The initial three levels facilitate the precise understanding of objects. We then comprehend the interaction between objects and the deep semantics embedded within the images. The fourth level centers on visual Object Relation (OR) detection, with the objective of identifying the basic relations between two objects (e.g., predict a walk-on relation between horse and grass). In the final level, the focus is on high-level comprehension of the images such as Event (E). Beyond the perceptual tasks, visual event detection requires VLMs to provide a summary of what is happening in the images and the participants involved in the event (e.g., the horse is eating grass). We highlight the flexibility of our curriculum pyramid that can be easily extended to incorporate new structural information at different levels with the goal of improving overall multimodal knowledge acquisition. This adaptability is facilitated by the flexible code-vision representations employed. Replay Buffer. We aim to empower VLMs with the capacity to proficiently acquire and maintain structural knowledge across various levels, aligning with the underlying motivation of lifelong learning research (Biesialska et al., 2020; Wang et al., 2023a). Although the acquisition of lower-level knowledge remains vital for higher-level knowledge attainment, directly training VLMs on highlevel tasks can lead to catastrophic forgetting, especially when dealing with infrequent and rare concepts. To prevent this, we build the replay buffer by random sampling at the level of the curriculum pyramid. During training, VLMs utilize the 13346 data samples specific to their respective level (i.e., samples from previous stages stored in the replay buffer). The amount of data we sampled from each stage can be found in Table 7 in the Appendix. 3.3 ViStruct Suite Construction We assemble a comprehensive dataset collection consisting of both labeled and unlabeled datasets for ViStruct corresponding to each level in the curriculum pyramid. The selected datasets along with their statistics are listed in Table 7 in the Appendix. To curate the object recognition dataset, we exclude rare classes with less than 500 samples and retain a total of 10,450 categories. On top of it, we add object grounding with 80.6M categories, object attributes with 14.6M attribute annotations, and object relations, using the resources in Table 7. For visual event understanding, we propose a weaklysupervised method to generate training data from the large-scale image-caption pairs in Table 7. As shown in Figure 4, for each image, we leverage the off-the-shelf semantic role labeling model1 to perform semantic parsing on the paired caption. Aligning with ViStruct, we adopt the FrameNet for the event and corresponding argument definitions. More details are included in Appendix E. We gather the aforementioned datasets and conduct a subsequent post-processing procedure. To establish consistency with ViStruct across various datasets, we align all categories from different datasets to the WordNet and FrameNet synsets through the automatic mapping process implemented in the NLTK toolkit (Bird et al., 2009). In cases where multiple categories in a dataset are assigned to the same synset, we manually adjust the mapping based on our prior knowledge. 3.4 Model Training ViStruct is based on continued pretraining, and can work on any pre-trained VLMs. In this work, we adopt OFA-base (Wang et al., 2022a) as our backbone model since it offers a simple seq-to-seq modeling approach. We train the model by feeding images i and the masked sequence mask(s) as defined in Equation 3.2 as input to the encoder, and train the decoder to produce s conditioned on the encoder outputs. Before the curriculum training, we include a warm-up pretraining stage to facilitate the models’ acquisition of fundamental code syntax structures and visual concepts with their definitions (see 1 https://github.com/chanind/ frame-semantic-transformer Appendix C). For the initial three levels in the curriculum, the model is trained for one epoch, while for the last two levels involving deep-semantic understanding, the model undergoes three epochs of training. Specifically, we introduce a focusing optimization trick that only optimize loss for the masked tokens (see Appendix D for more details). 4 Experiment We evaluate ViStruct on three structure prediction tasks, including visual relation detection (Yu et al., 2023), scene graph classification (Zellers et al., 2018; Yao et al., 2021), and situation recognition (Yatskar et al., 2016, 2017). We refer to Appendix A for implementation details. 4.1 Visual Relation Detection Dataset & Metrics. We evaluate the visual relation detection task on Visual Genome (Krishna et al., 2017). Following Krishna et al. (2019), we adopt the refined version that only contains 20 welldefined relation categories. Each image is annotated with identified objects and their relations. The task is to detect the relation between two given objects (provided along with their bounding boxes). For evaluation metrics, we measure two types of model performance following previous work (Yao et al., 2021). First, we report the micro-averaged recall in top K relation predictions (R@K). Second, we evaluate the marco-averaged recall of all 20 relation types across the top K predictions (mR@K), intending to measure the performance on long-tail relations. In our study, we select K=50, 100. Baselines. We compare ViStruct against three types of baseline methods. (1) Task-specific models: We consider the Neural motifs (Zellers et al., 2018) (Motif) that incorporates task-specific designs (e.g., off-the-shelf object detectors); (2) Extra supervision-enhanced methods: We consider the distant supervision method proposed in Yao et al. (2021), which introduces extra signals for pretraining (Motif+P) and denoising (Motif+D); (3) Ablations of ViStruct: We aim to measure the separate gains of two designs within ViStruct. Specifically, we quantify the gains of code-vision representations by measuring the performance of the backbone model OFA-base, which is trained on the same datasets but with natural language (i.e., image-conditioned language generation) (ViStructText). In assessing curriculum learning ablation, 13347 Method R@50 R@100 mR@50 mR@100 Mean Method R@50 R@100 mR@50 mR@100 Mean Motif* Motif+P* Motif+D* ViStruct-Text ViStruct-Mix ViStruct 67.93 73.22 76.28 73.39 74.71 75.71 70.20 75.04 77.98 75.16 76.32 78.53 52.65 60.44 60.20 63.27 61.41 63.71 55.41 63.67 63.61 67.42 68.26 70.71 61.55 68.09 69.52 69.81 70.10 72.17 Motif* Motif+P* Motif+D* ViStruct-Text ViStruct-Mix ViStruct 31.14 34.11 35.93 30.55 32.34 34.72 31.92 34.88 36.47 31.04 33.65 35.27 23.53 26.51 28.07 26.26 26.22 30.51 25.27 27.94 30.09 28.73 27.87 32.26 27.97 30.86 32.64 29.15 30.02 33.19 Table 3: The results (%) of the visual relation detection. * denotes results from Yao et al. (2021). Table 4: The results (%) of the scene graph classification. * denotes results from Yao et al. (2021). we measure the performance of pre-training on the ViStruct Suite mixture (ViStruct-Mix). 4.3 Experimental Results. As shown in Table 3, ViStruct consistently improves the visual relation detection performance regarding both the common and long-tail relation types. The reason lies in the diverse relation types ViStruct encounters during the structural knowledge learning phase, which prevents it from overfitting to some frequent relation types. Additionally, we demonstrate the separate gains of two novel designs in ViStruct. 4.2 Scene Graph Classification Dataset & Metrics. We continue to utilize the refined Visual Genome dataset for evaluation purposes. The scene graph classification task involves predicting object categories and relation types based on bounding box annotations, thereby assessing the models’ concept recognition capabilities besides the ability to detect object interactions. We measure the R@K and mR@K as described in Sec. 4.1, and the correctness of the predicted relation is determined solely by the accuracy of the predicted relation types and object categories. Baselines. We adopt the same baselines Sec. 4.1. Experimental Results. As shown in Table 4, ViStruct consistently outperforms baseline methods. Additionally, the performance gap between ViStruct and baseline methods including taskspecific models and two ablations increases compared to the results in Table 3. This can be attributed to (1) the extensive coverage of fundamental visual concepts in ViStruct, aligning with the requirements of scene graph classification for object category identification, and (2) the carefully arranged curriculum that guides the model to progressively comprehend the visual structures, preventing the model from overfitting on the large dataset of visual concepts. Situation Recognition Dataset & Metrics. We evaluate ViStruct on SWiG dataset (Pratt et al., 2020). Each image in the dataset is annotated with the major activity and corresponding participants (a.k.a., objects). The models must sequentially predict both the activity and the participants involved. Following Cho et al. (2022), we measure whether the gold-standard activity is predicted in the top-1 (Top-1 Predicted Verb) and top-5 (Top-5 Predicted Verb) model predictions. The evaluation of participant prediction entails two aspects: Value-all examines whether all predicted participants for an activity are correct, while Value evaluates the accuracy of each predicted participant in their assigned role. Participant prediction is considered correct if it matches any of the ground truth annotations. All metrics are calculated on a per-verb-specific basis and subsequently averaged across all verbs. Baselines. We compare ViStruct against three types of baseline methods. (1) Traditional taskspecific models: We include GraphNet (Li et al., 2017), CAQ w/ RE-VGG (Cooray et al., 2020), and Kernel GraphNet (Suhail and Sigal, 2019), as studied in Cho et al. (2022); (2) State-of-the-art task-specific models based on Transformers: We consider the CoFormer model that consists of two interactive modules (Cho et al., 2022). (3) Ablations of ViStruct: We consider the same ablations of ViStruct introduced in Sec. 4.1. Experimental Results. As shown in Table 5, ViStruct demonstrates substantial enhancements in both activity and participant detection, which can be attributed to its weakly-supervised training approach utilizing web-scale data. Our approach effectively utilizes the captions to generate visual event structures depicted in the web images. The benefit lies in the increased inclusivity of infrequent events and visual concepts, facilitated by the extensive volume of image-caption pairs. In addi- 13348 Metric Set Top-1 Predicted Verb Top-5 Predicted Verb Ground-Truth Verb Method verb value value-all verb value value-all value value-all Dev GraphNet* CAQ w/ RE-VGG* Kernel GraphNet* CoFormer* ViStruct-Text ViStruct-Mix ViStruct 36.93 37.96 43.21 44.41 44.10 44.68 46.23 27.52 30.15 35.18 35.87 35.04 35.58 38.82 19.15 18.58 19.46 22.47 21.19 21.73 24.29 61.80 64.99 68.55 72.98 71.04 71.15 74.81 45.23 50.30 56.32 57.58 55.24 55.65 60.91 29.98 29.17 30.56 34.09 31.76 32.61 37.41 68.89 73.62 73.14 76.17 75.04 75.31 79.23 41.07 38.71 41.68 42.11 40.18 40.82 46.23 Test GraphNet* CAQ w/ RE-VGG* Kernel GraphNet* CoFormer* ViStruct-Text ViStruct-Mix ViStruct 36.93 37.96 43.21 44.41 44.94 44.72 46.89 27.52 30.15 35.18 35.87 35.86 35.60 38.58 19.15 18.58 19.46 22.47 21.99 21.77 24.91 61.80 64.99 68.55 72.98 71.19 71.03 74.57 45.23 50.30 56.32 57.58 55.70 55.51 60.40 29.98 29.17 30.56 34.09 32.69 32.62 37.53 68.89 73.62 73.14 76.17 75.29 75.23 78.99 41.07 38.71 41.68 42.11 40.83 40.81 46.62 Table 5: The results (%) of the situation recognition task. * denotes results from Cho et al. (2022). Accumulative Improvement Across Curriculum Levels 75 R@50 R@100 mR@50 mR@100 Mean ViStruct-Text ViStruct 7.05 18.22 10.23 24.43 3.02 6.66 4.43 8.89 6.18 14.55 Table 6: The results (%) of the zero-shot visual relation detection task. 70 65 tion types. This demonstrates the benefits of the curriculum learning paradigm for generalization in low-resource settings. R@50 R@100 mR@50 mR@100 60 0 1 2 3 Curriculum Level 4 5 Figure 5: The effectiveness of the curriculum learning framework for visual relation detection. The metrics are introduced in Sec. 4.1. The dotted lines denote the results of ViStruct-Mix. tion, we observe the concrete gains associated with the utilization of curriculum learning. In alignment with the conclusion in Sec. 4.2, curriculum learning aids the VLMs to focus on high-level visual structure comprehension in the later stages, while simultaneously mitigating the risk of overfitting. 5 Method Further Analysis 5.2 Zero-shot Capability We evaluate ViStruct on the zero-shot visual relation detection task. The results are presented in Table 6. We compare with the results of training on natural language (ViStruct-Text) introduced in Sec. 4.1, and show that the code-vision representation exhibits significantly better zero-shot performance (i.e., 2.4x better on average). The codevision representation naturally facilitates the zeroshot visual structure prediction ability, owing to the unified format in both the pre-training and the downstream tasks. 5.1 Curriculum Learning 6 We conduct an ablation study to measure the performance of models trained at each stage of the curriculum on the visual relation detection task. We also compare the results of directly pre-training on the mixture of ViStruct Suite (ViStruct-Mix). As shown in Figure 5, as the training progresses, the evaluation metrics regarding two aspects continue to increase, gradually surpassing the performance of ViStruct-Mix, especially for the long-tail rela- We present a visual structural knowledge extraction framework ViStruct as a new step towards multi-granularity semantic parsing of visual data. First, we utilize the inherent structured of programming language to represent visual structural knowledge of different levels in a unified way. To capture the interdependencies among various granularities, we then leverage linguistic cues and propose a curriculum learning pyramid 13349 Conclusion that guides the models to effectively acquire visual structural knowledge of each level. Results show significant and consistent improvements on structure prediction tasks across all levels. Limitations and Future Work This paper utilizes the programming language to represent visual structures, with a focus on knowledge representation. However, due to the constrained capability of the base model, direct code generation is not feasible, resulting in reduced performance in downstream tasks. One potential solution is to integrate more advanced vision-language models, such as BLIP-2 with more capacity. By doing so, the trained models can effectively generate code representations of visual structures for diverse images in downstream tasks. Acknowledgement We thank the anonymous reviewers for their suggestions and comments. This research is based upon work supported by U.S. DARPA ECOLE Program No. HR00112390060 and U.S. DARPA KAIROS Program No. FA8750-19-2-1004. The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of DARPA, or the U.S. Government. 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Association for Computational Linguistics. 13355 A Adaptation of Downstream Tasks All evaluations are conducted within the supervised learning paradigm, involving training models using the training dataset, and selecting superior models as dictated by key performance indicators in the validation dataset. The ultimate performance of these models is evaluated on the testing dataset. We enumerate the specifics of adapting to downstream tasks as detailed below. A.1 Visual Relation Detection In this task, the models are provided with groundtruth annotations of the object bounding boxes and their respective categories. The requirement is for these models to effectively predict the visual interrelations between each pair of objects in the images. In our implementation, each object pair is supplied with their respective annotated bounding boxes, in conjunction with a mask prompt as input parameters for the ViStruct. ViStruct is then tasked with predicting the visual relation between the specified objects. Assuming the pair of objects provided are a horse and grass, the inputs introduced to ViStruct would be as follows: we input the mask prompt with annotated location to ViStruct: # Generate Concepts object1 = <mask>( location= [bounding_box], attribute=None ) ViStruct is then trained to predict the ground truth object categories, for example, the “horse”. The visual relation detection task is trained in the same vain as in Sec. A.1. A.3 Situation Recognition In this task, the models are required to predict the visual event (a.k.a, the activity) in the image with corresponding participants. We treat it as two different tasks, namely the visual event prediction and the event role prediction. For the visual event prediction, we input the prompt as: # Generate Visual Event with Arguments ViStruct is trained to predict the visual event, for example, “eat”. For the event role prediction task, the models are provided with the ground-truth or predictive visual event with a mask prompt as input: # Generate Concepts object1 = horse( location=[bounding_box],# Generate Visual Event with attribute=None Arguments eat(ingestor=<mask>) ) ViStruct is then trained to predict the object for object2 = grass( the argument “ingestor”. Specifically, in this task, location=[bounding_box],we perform an automatic mapping to convert each attribute=None potential visual event and object to a single token ) in the OFA model for training and evaluation. The # Generate Relations automatic mapping is based on the word embed<mask>(sub=object1, obj=object2) ding of the OFA model. We select the most similar Correspondingly, ViStruct is trained to predict the ground truth relation: “walk_on” as output. We meticulously select and refine appropriate verbalizers for every unique relation to ensure that each relation within the label space can be represented by a single, meaningful token in the OFA model. A.2 Scene Graph Classification In this task, the models are provided with the bounding boxes and are required to forecast both the classifications of the objects and their corresponding visual relations. We treat it as a multitask learning problem, training ViStruct to perform both the object recognition task and the visual relation detection task. For the object recognition task, token, measured by the cosine similarity, with the target verbalizers of visual events and objects. B Visual Structure Learning Several studies have been conducted consecutively on learning multimodal structural knowledge, commencing with visual concept recognition as a basic task (Deng et al., 2009; Davoodi et al., 2023; Kamath et al., 2022), progressing to object grounding (Yu and Ballard, 2004; Shao et al., 2019; Krasin et al., 2017), object attribute detection (Chen et al., 2023a; Patil and Abhyankar, 2023; Bravo et al., 2022), object-object relation detection (Krishna et al., 2016), and finally visual event understanding (Yatskar et al., 2016, 2017; Cho et al., 2022; 13356 Type Pretraining Task Source #Sample #B_Sample Supervised Object Recognition Object Grounding Attribute Detection Relation Detection ImageNet21K Object365, OpenImages, COCO LSA OpenImages, Visual Genome 1M 2.5M 0.5M 0.5M 104K 1M 0.1M 0.1M Unsupervised Event Detection COCO, SBU, CC3M 4M - Table 7: The datasets included in ViStruct Suite. #B_Sample denotes the number of samples in the replay buffer. E Figure 6: The focusing optimization trick prioritizes the semantic content of generated code (highlighted portion) while disregarding the code’s syntactic structure. Li et al., 2020b, 2022b). In this work, we design a curriculum learning pyramid that progressively teaches VLMs the aforementioned structural knowledge and assemble a dataset collection that encompasses multi-level structural knowledge. C Warm-up Pretraining Prior to engaging in multimodal structural knowledge learning, a warm-up stage is implemented to facilitate the models’ acquisition of fundamental code syntax structures and visual concepts with their definitions. To accomplish this, we integrate code training data (Kocetkov et al., 2022), code-formulated class definitions of WordNet and FrameNet synsets, and all datasets within the ViStruct Suite, then subject the models to a single training epoch. D Focusing Optimization Through our analysis, we observe that the code output contains redundant information regarding the syntax structure, which can be quickly learned during the warm-up stage. Consequently, we propose to focus solely on the semantic content of the generated code, meaning that the loss computation will be limited to the semantic content exclusively. As depicted in Figure 6, this approach can direct models to prioritize structural knowledge and accelerate the training process. Details of ViStruct Suite The selected datasets along with their statistics are listed in Table 7. For object recognition, we utilize the pre-processed ImageNet-21K (Ridnik et al., 2021). To curate the dataset, we exclude rare classes that consist of less than 500 samples and retain a total of 10,450 categories, each with a sample size of 100. For object grounding, we leverage object detection datasets, including COCO (Lin et al., 2014), OpenImages (Krasin et al., 2017), and Object365 (Shao et al., 2019), which contain 80, 600, 365 categories respectively. For object attribute detection, we adopt the open-vocabulary LSA (Pham et al., 2022) that contains 14.6M attribute annotations in total. For visual object relations detection, we adopt the OpenImages (Krasin et al., 2017) and Visual Genome (Krishna et al., 2016). We adopt the training split of these labeled datasets to avoid data contamination. For visual event understanding, we propose a weakly-supervised method to generate training data from the large-scale image-caption pairs in COCO (Lin et al., 2014), SBU Captions (Ordonez et al., 2011), and CC3M (Sharma et al., 2018). As shown in Figure 4, for each image, we leverage the off-the-shelf semantic role labeling model2 to perform semantic parsing on the paired caption. Aligning with ViStruct, we adopt the FrameNet for the event and corresponding argument definitions. We gather the aforementioned datasets and conduct a subsequent post-processing procedure. To establish consistency with ViStruct across various datasets, we align all categories from different datasets to the WordNet and FrameNet synsets through the automatic mapping process implemented in the NLTK toolkit (Bird et al., 2009). In cases where multiple categories in a dataset are assigned to the same synset, we manually adjust the mapping based on our prior knowledge. 2 https://github.com/chanind/ frame-semantic-transformer 13357
https://openalex.org/W2027969104	https://ubibliorum.ubi.pt/bitstream/10400.6/9244/1/O%20ethos%20jornal%c3%adstico_%20da%20t%c3%a9cnica%20%c3%a0%20reflex%c3%a3o%20cr%c3%adtica%20_%20Correia%20_%20Verso%20e%20Reverso.pdf	Portuguese	null	O ethos jornalístico: da técnica à reflexão crítica	Verso e Reverso	2,009	cc-by	8,258	O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso 10/02/2020 10/02/2020 CAPA SOBRE ACESSO CADASTRO PESQUISA ATUAL ANTERIORES NOTÍCIAS SUBMISSÃO POLÍTICA DE ÉTICA INDEXAÇÃO CALL FOR PAPERS CONTATO Capa > v. 23, n. 54 > Correia doi: 10.4013/ver.2009.23.54.03 O ethos jornalístico: da técnica à reflexão crítica The journalistic ethos: from technique to a critical reflexion João Carlos Correia jcfcorreia@gmail.com Resumo. Durante décadas, o jornalismo foi entendido como uma simples prática profissional (uma ocupação), que poderia existir sem a necessidade de uma formação universitária especializada. Neste texto defende-se a formação universitária como a mais adequada para os desafios e exigências da profissão, recordando-se que a mesma implica uma distância crítica em relação às condições concretas do seu exercício. Desta forma defende-se que o ethos jornalístico, baseado numa atitude pragmática em relação à realidade, terá de se confrontar também com as exigências críticas que caracterizam um saber reflexivo teoricamente orientado. Esta problemática recorrentemente retomada em diversos debates (ensino profissionalizante versus saber universitário, teoria versus prática, capacidades reflexivas versus competências performativas) relaciona-se directamente com uma reflexão sobre a natureza da profissão. Abstract. For decades, journalism was seen as a simple practice (an occupation), which could exist without the need for specialized academic training. In this paper we defend the university as the most suitable way for the challenges and demands of the profession, remembering that it implies a critical distance in relation to the concrete conditions of professional practice. Thus it is argued that the journalistic ethos, based on a pragmatic attitude towards reality, will deal also with the critical requirements that characterize a knowledge theoretically oriented. This problem repeatedly taken up in various discussions (vocational versus academic knowledge, theory versus practice, reflective capabilities versus performative skills) is directly related with a reflection on the nature of the profession. Introdução Durante décadas, o jornalismo foi entendido como uma simples prática profissional (uma ocupação), que poderia existir sem a necessidade de uma formação universitária especializada. Esta situação permaneceu, em numeroso países, onde USUÁRIO Login Senha Lembrar usuário Acesso CONTEÚDO DA REVISTA Pesquisa Todos Pesquisar Procurar Por Edição Por Autor Por título Outras revistas IDIOMA Português (Brasil TAMANHO DE FONTE CONTEÚDO DA REVISTA O ethos jornalístico: da técnica à reflexão crítica The journalistic ethos: from technique to a critical reflexion e poderia sitária ncia do numa am um ed for Pesquisa Todos Pesquisar Procurar Por Edição Por Autor Por título Outras revistas João Carlos Correia jcfcorreia@gmail.com João Carlos Correia jcfcorreia@gmail.com Resumo. Durante décadas, o jornalismo foi entendido como uma simples prática profissional (uma ocupação), existir sem a necessidade de uma formação universitária especializada. Neste texto defende-se a formação univ como a mais adequada para os desafios e exigências da profissão, recordando-se que a mesma implica uma dis crítica em relação às condições concretas do seu exercício. Desta forma defende-se que o ethos jornalístico, bas atitude pragmática em relação à realidade, terá de se confrontar também com as exigências críticas que caracte saber reflexivo teoricamente orientado. Esta problemática recorrentemente retomada em diversos debates (ens profissionalizante versus saber universitário, teoria versus prática, capacidades reflexivas versus competências performativas) relaciona-se directamente com uma reflexão sobre a natureza da profissão. Abstract. For decades, journalism was seen as a simple practice (an occupation), which could exist without the specialized academic training. In this paper we defend the university as the most suitable way for the challenge demands of the profession, remembering that it implies a critical distance in relation to the concrete conditions professional practice. Thus it is argued that the journalistic ethos, based on a pragmatic attitude towards reality also with the critical requirements that characterize a knowledge theoretically oriented. This problem repeatedly in various discussions (vocational versus academic knowledge, theory versus practice, reflective capabilities ver performative skills) is directly related with a reflection on the nature of the profession. Capa > v. 23, n. 54 > Correia Capa > v. 23, n. 54 > Correia doi: 10.4013/ver.2009.23.54.03 CAPA SOBRE ACESSO CADASTRO PESQUISA ATUAL ANTERIORES NOTÍCIAS SUBMISSÃO POLÍTICA DE ÉTICA INDEXAÇÃO CALL FOR PAPERS CONTATO Capa > v. 23, n. 54 > Correia doi: 10.4013/ver.2009.23.54.03 O ethos jornalístico: da técnica à reflexão crítica The journalistic ethos: from technique to a critical reflexion Introdução xistir onde TAMANHO DE FONTE nte décadas, o jornalismo foi entendido como uma simples prática profissional (uma ocupação), que pode a necessidade de uma formação universitária especializada. Esta situação permaneceu, em numeroso país 1/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso hoje se continua a dispensar a exigência de qualquer diploma e onde se continua a fazer uma reflexão sobre qu caminhos mais adequados sobre essa formação. Tal situação contrasta de forma acentuada com a proposta sub Lee Bollinger, presidente da Columbia University que, ao propor a reforma dos estudos jornalísticos na prestigia Graduate School of Journalism daquela Universidade, defendeu a necessidade de um equilíbrio entre o ensino d jornalismo como profissão e uma formação académica reflexiva, historicamente situada e teoricamente orientad que neste texto esta opção se afigura como aquela que se considera mais adequada para os desafios e exigênci profissão, importa recordar que a mesma implica uma distância crítica em relação às condições concretas de ex profissão. Gera-se, deste modo, mais uma vez, um dilema dirigido ao ethos jornalístico: um ethos baseado num pragmática em relação à realidade terá de se confrontar também com as exigências críticas que caracterizam u reflexivo teoricamente orientado. Grande parte deste dilema, travestido de diversas formas (ensino profissional versus saber universitário, teoria versus prática, capacidades reflexivas versus competências performativas) est em muitos dos debates sobre as necessidades formativas do jornalismo e acaba por se relacionar directamente questionamento sobre a natureza da profissão. ssima Sendo da cício da atitude saber nte presente om o uída e, e teoria ? Que ? Que INFORMAÇÕES Para leitores Para Autores Para Bibliotecários OPEN JOURNAL SYSTEMS Ajuda do sistema Sendo da ício da atitude saber nte resente INFORMAÇÕES Para leitores Para Autores Para Bibliotecários A problemática da estandardização de uma base cognitiva comum para a formação de profissionais não está co por isso, continuam-se a formular e a renovar certas perguntas: “O que é jornalismo? Em que consiste o trabal fundamental do jornalista? O que é que um jornalista precisa de saber para poder desempenhar a sua função? do jornalismo existe, ou deveria ser desenvolvida, que possa servir de base ao ensino profissional desta discipli tipo de formação é a mais adequada para proporcionar esta aprendizagem? Académica ou profissional/vocacion outras disciplinas académicas podem contribuir para o ensino do jornalismo? Introdução Como se articulam essas disciplina matéria central do jornalismo?” (French, 2006, p. 464). Conhecem-se as condições que permitem o desenvolvimento de um campo de saberes autónomo numa área científica de estudo considerada necessária ao desenvolvimento de uma profissão especializada: ao lado de uma delimitação epistemológica e da identificação concreta de uma função social decorrente da partilha dos saberes, converge a necessidade de factores como a criação de escolas, institutos e faculdades, o desenvolvimento de fundações especializadas, a emergência de um corpo de académicos e investigadores, o reconhecimento da importância ou da necessidade desses estudos superiores no acesso à profissão, a existência de sociedades científicas próprias, a existência de editoras, revistas e publicações e a manutenção de um relacionamento estável entre a comunidade científica e a comunidade dos profissionais. Não se pode dizer que este percurso nem a confluência destes factores favoráveis tenha sido seguido ou verificado no jornalismo, ao menos de um modo linear e com idêntica persistência e intensidade em todas as latitudes. Portugal, por exemplo, seguiu um percurso diferente do Brasil nomeadamente no que respeita à importância dos estudos universitários e, particularmente, no que diz respeito ao reconhecimento do título enquanto condição de acesso à profissão. Para isso contribuiu de modo decisivo a inexistência de um relacionamento frutuoso entre a indústria jornalística e a academia. Não existiu “uma prática de diálogo continuado e totalmente isento de ideias preconcebidas, que permitisse “compreender e intervir no fosso entre o universo académico e o universo profissional” (Pinto, 2004, p. 55). Um pouco por todo o lado, verificaram-se acidentes na profissionalização do jornalismo. Apesar do papel central conferido aos jornalistas pela teoria da democracia, bem identificada na célebre metáfora do “Quarto Poder”, o jornalismo tem sido, historicamente, uma profissão pouco prestigiada (Traquina, 2004, p. 27). Segundo Schudson (citado por Traquina, 2004, p. 27), apesar de “os jornalistas na Europa e nos Estados Unidos e noutros locais se terem associado aos ricos e poderosos, nunca foram respeitados por eles”. Para esse facto, contribuiu certamente a antiga aura de uma profissão que funcionava para pessoas que não tinham conseguido vingar noutros ofícios, em regime de duplo emprego ou olhada como um passo para atingir um cargo politico, não dispondo, por isso, de uma imagem de uma profissão por direito próprio (Traquina, 2004, p. 28-30). O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso contrário do que se verificou noutras profissões, os caminhos que permitem aceder ao seu desempenho. A definição do jornalista profissional como alguém que desempenha esta actividade como ocupação principal, a tempo inteiro e remunerada em publicações periódicas, retirando daí o essencial dos seus recursos – uma definição surgida na Franca e vigente em Portugal – faz com que a definição de jornalismo não se identifique com o desempenho de determinadas competências, mas pelo exercício da actividade num jornal: é jornalista quem obtém emprego numa organização jornalística, assim os mecanismos de acesso à profissão transferem-se do ambiente cultural, científico e profissional para o âmbito socioeconómico (Fidalgo, 2008, p. 73). Assim, enquanto o engenheiro deveria estudar engenharia, o arquitecto, arquitectura, e o kurista, direito e leis, pensou-se, por vezes, que o jornalista não precisaria de estudar nada de específico, embora se lhe conferisse alguma aura humanística que se traduzia na ideia generalizante e bem pouco definida de que devia ser culto ou, de uma forma mais simplificada, devia ser “possuidor de bagagem”. Baseadas na comparação com o direito e a medicina, as teorias que analisam o profissionalismo, consideram que uma profissão para existir tem que ter um controlo sobre a base cognitiva da profissão. A característica que melhor define uma profissão é o conhecimento abstracto controlado por grupos profissionais pois só um conhecimento governado por abstracção permite dimensionar novas problemas e novas tarefas. Nesse sentido, é também um recurso estratégico essencial de afirmação da legitimidade da profissão. Na aquisição, aprofundamento e certificação desse conhecimento abstracto, acaba por ter papel essencial o conhecimento académico que tem uma função óbvia de legitimação do trabalho dos profissionais e ajuda ao desenvolvimento de novos diagnósticos e tratamento de problemas específicos. Assim, a existência de uma formação científica em escolas especializadas parece ser um dos critérios que deve ser preenchido para o aparecimento de uma profissão (Fidalgo, 2008, p. 38, 41). Com efeito, a aparição e diferenciação de um campo específico implica a aparição de um grupo de profissionais que dominam uma linguagem especializada e instauram uma diferença em relação aos “não profissionais”. Neste sentido, uma especialização significa autoridade e autonomia (Traquina, 2004, p. 16). Os problemas do jornalismo são o sem número de caminhos organizacionais que podem conduzir à carreira e a ausência de estandardização da base cognitiva necessária para o exercício da profissão. Introdução O Dictionnaire Français des Professions (1880), à semelhança de reputadas enciclopédias, comenta: “O jornalismo não é uma profissão no sentido habitual da palavra. É-se engenheiro mesmo quando se não exerce; é-se médico ou advogado mesmo quando não existem clientes. Mas não se é jornalista a não ser quando se escreve no jornal; deixa-se de o ser ao outro dia. Não existe diploma nem certificado. O jornalismo não é uma profissão no sentido habitual do termo”. O mesmo Dicionário (citado por Traquina, 2004, p. 37) diz ainda: “(…) o que constitui uma profissão é que uma pessoa a prepara e que terminada esta preparação, toma-a. No jornalismo, não existe aprendizagem, nem diploma nem certificado. Basta haver o acordo do chefe de redacção”. A diversidade de circunstâncias e a indeterminação que percorre o acesso à profissão deve-se a um particular desenvolvimento e autonomia do campo jornalístico, o qual foi incapaz de uniformizar, ao 2/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Tais problemas implicam questões de reconhecimento/auto-reconhecimento, de legitimidade e de afirmação do seu poder. Nesse sentido, procede-se a discussão ainda não encerrada em diversos países acerca de quem pode ser jornalista. A afirmação de que não se é jornalista quando se escreve no jornal pode ter como corolário simétrico a conclusão de que basta escrever no jornal para ser jornalista (Correia, 1998, p. 7-8). Esta consideração preocupante terá sempre acompanhado a questão da legitimidade da profissão: um dos riscos é que, numa sociedade extremamente complexa, uma atitude desse género dificilmente proporciona a emergência consistente de um saber reflexivo, isto é, um saber que questione o saber estabelecido e, particularmente, interrogue as rotinas solidificadas. Ora, este tipo de saber – que questiona a própria rotina que se instala nas organizações burocráticas em que se pratica a profissão é, provavelmente, um dos mais decisivos, tendo em conta a pluralidade de desafios que se colocam na profissão. Outro risco é o de tornar a discussão sobre a profissão, a gestão dos seus conflitos e a definição dos seus instrumentos conceptuais e das suas práticas, algo que se torna erosivo para a própria identidade da profissão por ser discutido em instâncias de poder e de saber que lhe são alheias. Ora, qualquer profissão, no processo estratégico da sua afirmação, necessita de uma certa dose de auto-referencialidade, isto é da construção de critérios próprios legitimamente aceitos. Academia e indústria: equívocos e desencontros Grande parte do escasso desenvolvimento dos estudos jornalísticos, que durante anos se verificou em Portugal, foi acompanhado por uma intensa falta de diálogo entre a academia e as associações representativas dos profissionais, que se traduziu em momentos de alguma tensão, em que tais organizações pareceram claramente afastar-se de qualquer exigência clara no que respeita ao desenvolvimento de estudos de nível universitário. “Do lado da academia, proposições do tipo “não há nada mais prático do que uma boa teoria” ou “a teoria é mais importante do que a prática” constituem argumentos que servem, frequentemente, para acentuar a dissociação existente entre as duas esferas. Por outro lado, proposições como aquela que repete que “os cursos de jornalismo são excessivamente teóricos e desfasados do universo profissional”, com a ideia associada de que “a verdadeira escola de jornalismo é a experiência e a tarimba”, constituem o outro extremo que dificulta o desenvolvimento de pontes de comunicação” (Pinto, 2004, p. 58)”. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso As circunstâncias que envolveram o extremar destas posições fizeram com que o jornalismo se mantivesse durante décadas confinado a um estatuto marginal sistemático no seio da academia sendo muitas vezes olhado como um espaço 3/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso fechado à reflexividade crítica do saber universitário, lugar de aprendizagem das técnicas que não equacionava a produção de investigação ou de saber novo enquanto elemento integrante das necessidades de formação. A prática profissional tornou-se o método privilegiado de aprendizagem e durante décadas acreditou-se que essa prática deveria exigir pouco mais do que o domínio de determinadas técnicas e um conjunto de normas deontológicas legitimadas entre os pares e demais sectores sociais. Ignorou-se, por completo, um olhar sobre o jornalismo que o reflectisse como campo de ensino especializado, remetendo- se o conhecimento jornalístico, no fundo, para um tipo de conhecimento empírico obtido por osmose nas redacções, dispensado de reflectir sobre as suas práticas, sobre o desenvolvimento de novas teorias e metodologias: os estudos jornalísticos foram olhados sempre com desconfiança por jornalistas profissionais que percorreram o seu caminho através de um saber de experiência feito e observavam de soslaio os que reivindicavam a necessidade da formalização de um conhecimento especializado de nível universitário. Escassas vezes se pensou na formação de um campo epistemológico especializado que identificasse, na prática jornalística, um objecto legítimo que implicasse o desenvolvimento de reflexões adequadas e de metodologias especificamente direccionadas. Uma parte considerável desta problemática resulta obviamente de uma ausência de relacionamento entre a profissão e a academia. A legitimidade de uma profissão assenta também numa formação longa controlada pela elite da profissão. A existência de escolas de medicina, de engenharia ou de direito está associada ao esforço dos seus profissionais que reconhecem aquelas instituições e as formas de ensino que lhe estão associadas como um produto da necessária estandardização da base cognitiva necessária à afirmação de uma profissão. No caso do jornalismo, acontece que, por vezes, como sucedeu em Portugal, o surgimento de escolas de formação não está associada ao esforço de profissionais mas de académicos que decidem aplicar domínios epistemológicos pré-existentes: Ciências Sociais e Humanas, Letras e Humanidades e Ciências da Comunicação. Obviamente, os profissionais sentem dificuldades em aceitar a distanciação crítica que qualquer saber universitário implica porque a mesma lhes é proposta a partir de fora, classificando-a, de forma generalizadora, como uma deriva abstracta e especulativa Os académicos acentuam a importância dessa distância porque é nela que se funda uma parte da sua legitimidade, relativizando a importância dos saberes que se fundamentam interiorização de meras práticas profissionais por estes dispensarem a sua intervenção. Debates e interrogações Debates e interrogações Em meados dos anos 90, os participantes do 3º Congresso dos Jornalistas Portugueses, confrontados com o acesso de numerosos jovens à profissão e com a sucessão geracional que se verificava no interior da classe, debruçaram-se sobre o problema e lançaram, um tanto tardiamente, grande parte do seu discurso sobre a identidade da classe e em especial dos temas: como se aprende a profissão? Quais as condições que é necessário preencher para que se possa dizer que se é oficial de tal ofício? Será que só é jornalista quem escreve nos jornais? Como é que se forma um jornalista? Se é a situação profissional que o define – exercida a tempo inteiro e remunerada, como ocupação principal – então será que todos os que escrevem nos jornais e deles retiram os seus proventos maioritários são, por definição, jornalistas? Ou, pelo contrário, ser jornalista pressupõe a adopção de um certo estilo e de técnicas narrativas e de recolha de dados, de metodologias, de princípios e de saberes específicos que impliquem a padronização de um corpo de conhecimentos especializado? De uma forma talvez caricatural, pode-se dizer que, de um lado, se posicionaram os defensores da boa "tarimba", que acreditavam que o talento jornalístico não poderia ser ensinado nas academias já que a experiência, desvelada mãe de todos os saberes, forneceria os elementos essenciais aos profissionais para exercerem o seu mester com arte e sabedoria. Do outro, surgiam os académicos que afirmavam que sem uma cuidada preparação ética, deontológica, filosófica, sociológica, cultural e técnica, o jornalista não estaria preparado para exercer a sua profissão. De certa forma, a dependência dos Estudos Jornalísticos das Ciências Sociais, das Letras e das Humanidades gerou um efeito de desconfiança que produziu reflexos corporativos aos quais não foram alheios a presença continuada de um elevado número de profissionais com baixas habilitações literárias. Como realçam Mário Mesquita e Cristina Ponte (1996-1997), “a localização dos cursos nessas escolas teve, naturalmente, a consequência positiva de assegurar um sólido enquadramento no domínio das ciências sociais e humanas e uma garantia de autonomia perante tropismos corporativos que poderiam conduzir a um ensino puramente profissionalizante e empiricista. Em contrapartida, verificou-se um excessivo alheamento das problemáticas teórico-práticas do jornalismo e instaurou-se um clima de desconfiança mútua entre as instituições académicas e o meio profissional dos jornalistas.” 4/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 as.unisinos.br/index.php/versoereverso/article/view/5768/2 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso O que se tornou mais impressivo nalgumas das referências feitas no Congresso dos Jornalistas Portugueses em relação à formação exigida foi a exaltação de uma certa prática do jornalismo que remete para um "ontem" melhor do que “o hoje”. A aprendizagem por osmose na sala de redacção através do acompanhamento do chefe de redacção ou do colega de tarimba era colocada não apenas numa posição mais favorável do que a aprendizagem efectuada junto do docente universitário do Curso de Comunicação ou Jornalismo como, em última instância, com algumas excepções, era inclusive entendida como única e nem sequer complementar da outra. A discussão sobre a formação jornalística em Portugal é antiga. Do 1° Boletim do Sindicato dos Trabalhadores de Imprensa (1926) já constava a proposta de criação de uma Escola Superior de Jornalismo que, ao tempo, mereceu o seguinte comentário do director do jornal Comércio do Porto: "Da mesma forma que não há escola de poesia também não há escolas de jornalistas". Em 1941, o Sindicato Nacional de Jornalistas entregou um projecto de um Curso de Formação Jornalística, justificado pela necessidade de "promover, de maneira decisiva, a valorização profissional dos jornalistas bem como o seu nível de cultura para os limites exigidos para a missão que desempenha". Finalmente, em fevereiro de 1970, uma delegação de uma direcção sindical apresentou a proposta de criação do Ensino Superior de Jornalismo. A criação do ensino universitário de jornalismo identificava-se com uma reivindicação progressista em larga medida, com a defesa da própria liberdade de expressão e com alguns ventos de mudança que perpassavam então pela sociedade portuguesa e se reflectiam no campo jornalístico. Porém, só em 1979 surgiria, na Universidade Nova, a primeira licenciatura em Comunicação Social. No ano seguinte, surgiu outra licenciatura com idêntica designação no Instituto Superior de Ciências e Politicas da Universidade Técnica de Lisboa e a Universidade Católica Portuguesa iniciou os seus primeiros Cursos de Ciências da Informação configurados como pós-graduações destinadas a pessoas possuidoras de formação universitária ou a profissionais de jornalismo com mais de cinco anos de experiência. As propostas formativas emergentes entre finais dos anos 70 e 80 fizeram a sua aparição enquanto Cursos de Comunicação Social ou de Ciências da Comunicação integrados em escolas, faculdades ou institutos preferencialmente ligadas às ciências sociais e humanas, com uma acentuada valorização das componentes histórico-filosófica, linguística, sociológica e tecnológica (Pinto 2004, p. 51- 53). O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso profissional de jornalista, é condição fundamental para o exercício da profissão em Portugal, sendo emitido e renovado pela Comissão da Carteira Profissional de Jornalista (Artº 4º, ponto 1). No que concerne ao acesso ao título profissional, a única condição é o candidato ser maior de 18 anos e estar no pleno gozo dos seus direitos civis (Artº 2º). Em termos de habilitações literárias, verifica-se uma ausência de quaisquer referências a exigências podendo deduzir-se da lei geral que é exigida a escolaridade mínima obrigatória. A profissão inicia- se com um estágio obrigatório. Este tem “a duração de 12 meses, em caso de licenciatura na área da comunicação social ou de habilitação com curso equivalente, ou de 18 meses nos restantes casos”(Artº 5, ponto 1). Numa interpretação estrita dos termos legais, que enquadram o exercício da profissão, não se verificam quaisquer incentivos de monta para a busca de conhecimentos universitários como fundamento do saber jornalístico: a duração do tempo de estágio é a única diferença visível existente entre candidatos com níveis de habilitação tão distintos como a escolaridade mínima obrigatória e o mestrado. Recentemente, o Governo esteve disposto a propor a licenciatura como habilitação mínima de ingresso na profissão jornalística, mas esse ponto foi alterado devido, nomeadamente, à oposição do sindicato dos jornalistas. Apesar de tudo, 60,3% dos jornalistas é detentor deste grau de ensino, sendo 34% licenciados em comunicação ou jornalismo (Canavilhas, 2009). Desta forma, gera-se uma perplexidade que deve ser matéria de reflexão: o mercado de emprego sanciona de forma positiva a licenciatura em comunicação/jornalismo, mas a comunidade de representantes dos profissionais continua a manter dúvidas substanciais sobre essa necessidade. Transformações do jornalismo: desafios de formação Apesar de tudo, multiplicam-se as vozes no sentido de que haverá que encontrar formas de articular a formação académica com as exigências legais de acesso à profissão. Nesse sentido, há um reconhecimento por parte de um número crescente de jornalistas da necessidade de ultrapassar os sucessivos impasses que têm impedido essa articulação. Da mesma forma, regista-se um maior cuidado das academias em fornecerem uma formação especializada, a qual embora não se reduza, obviamente, a uma orientação estritamente profissionalizante, seja, todavia, capaz de produzir competências profissionais acompanhadas de saber reflexivo, sem se perderem na famosa deriva abstracta e especulativa. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso o início dos anos 80 surgem o Centro de Formação dos Jornalistas do Porto (1983) e o Centro Protocolar d ornalistas (Cenjor), expressão de um descontentamento com o ensino universitário do jornalismo. Finalmente, nos anos 90, surgem os cursos especificamente designados como de jornalismo, a explosão de cursos de jornalismo no ensino politécnico e, finalmente, o aparecimento e expansão de uma série de formações pós-graduadas em jornalismo. A partir de 2005 teve lugar o processo de revisão das carreiras universitárias de forma a adaptarem-se aos objectivos do Tratado de Bolonha, o qual visa a harmonização do Ensino Superior no espaço da União Europeia. Presentemente, existem em Portugal dez cursos universitários públicos, seis cursos universitários privados, nove cursos politécnicos públicos e nove cursos politécnicos privados na área de Ciências da Comunicação/Jornalismo. Há uma demarcação do ensino em jornalismo nalgumas licenciaturas e, sobretudo, no ensino pós – graduado (ao nível de mestrado). Verifica-se também o aumento da produção científica sobre o jornalismo, com linhas de investigação significativas e editoras particularmente direccionadas para jornalismo. Finalmente, o Grupo de Trabalho de Jornalismo e Sociedade da Sociedade Portuguesa de Comunicação destaca-se como o que maior número de inscrições de trabalhos regista nos congressos da SOPCOM. Apesar de o mercado de trabalho disponível começar a evidenciar uma preferência acentuada pelos graduados e pós– graduados em comunicação ou jornalismo e apesar de uma relativa maior aproximação entre a academia e o exercício da profissão, o actual Estatuto do Jornalista vigente em Portugal não identifica qualquer espécie de habilitação especial para se ser jornalista: “São considerados jornalistas aqueles que, como ocupação principal, permanente e remunerada, exercem com capacidade editorial funções de pesquisa, recolha, selecção e tratamento de factos, notícias ou opiniões, através de texto, imagem ou som, destinados a divulgação, com fins informativos, pela imprensa, por agência noticiosa, pela rádio, pela televisão ou por qualquer outro meio electrónico de difusão” (Artº 1º Ponto 1). São também considerados jornalistas “os cidadãos que, independentemente do exercício efectivo da profissão, tenham desempenhado a actividade jornalística em regime de ocupação principal, permanente e remunerada durante 10 anos seguidos ou 15 interpolados, desde que solicitem e mantenham actualizado o respectivo título profissional” (Artº 1º, ponto 3). Tal título, denominado carteira 5/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Da mesma forma, existe um número cada vez mais significativo de profissionais reconhecidos que começam a leccionar na Academia, fazendo os respectivos trajectos no jornalismo e no seu ensino. Esta situação evolui em simultâneo com a consciência da necessidade de pensar a profissão, num mundo onde se generalizam as permanentes mudanças e perplexidades que se constituem como desenvolvimento desta actividade profissional. Com efeito, o jornalismo desenvolve-se como um campo marcado por importantes transformações. Uma primeira transformação resulta do aproveitamento jornalístico da internet como meio de difusão, o qual conduziu a transformações nos restantes meios através, nomeadamente, da sua imigração para a rede. Simultaneamente, a internet configurou-se como uma ferramenta de recolha de dados e de informações, gerando, também, interactividade com fontes e receptores. Por outro lado, permitiu o aparecimento de actividades parajornalísticas (como a produção de blogs) que têm produzido um forte impacto no modo como se pensa a profissão. A internet transformou as condições de agendamento e de selecção da informação e destronou o jornalista do seu papel de detentor quase absoluto do monopólio dessa mesma selecção. Finalmente, aumentou a segmentação da informação e despertou a possibilidade de formas de consumo e de recepção cada vez mais personalizados (Sousa, 2004, p. 1-2). As novas tecnologias reconfiguram as linguagens, as técnicas e as rotinas: saberes de procedimento (como agir para conseguir uma notícia ou lidar com uma fonte?) e de narração (como se redige para a pluralidade de plataformas disponíveis: papel, internet, telemóveis, etc?) sofrem mutações importantes. A proliferação de plataformas e a segmentação de canais abrem o campo importante dos jornalismos especializados. A digitalização do sector é mais um desafio que solicita ao jornalismo e aos jornalistas um especial cuidado na compreensão do lugar de onde fala: a análise das condições de enunciação implica não apenas o nível tecnológico e aconselha que o jornalista não se transforme num McGyver alucinado que contempla gadgets resplandecentes.. Um certo mcluhanismo apressado tem servido de ideal justificador de uma subordinação da formação jornalística às transformações tecnológicas e às consequentes necessidades do mercado de emprego. Na época do capitalismo cognitivo, o pensamento e o estatuto dos intelectuais sofreu uma mudança radical constatando-se um fenómeno de proletarização que fere as universidades, o ensino, os professores e os media. (Correia, 2007, p. 11). O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso de competências reflexivas. Esta ênfase nas competências performativas segue, muitas vezes, um percurso unilateral e redutor que, no caso do jornalismo, é justificado ideologicamente pela mudança de paradigma introduzido pelas novas tecnologias. No limite, a formação jornalística é uma questão de aprender a usar ferramentas para produzir conteúdos. Neste sentido, há toda uma concepção de profissão que se alimenta da ideologia tecnocêntrica. É evidente que é necessário saber como funcionam os aparelhos, mas interessa não perder de vista para que servem, quando se utilizam e porque se utilizam. O problema da mediação deve ser questionado na sua natureza, nas relações sociais que o determinam e configuram, no que respeita aos processos de relacionamento social que originam, quanto às transformações que produzem no desempenho da profissão. Embora ser polivalente no desempenho de tarefas e ser dotado de competências multimediáticas seja para qualquer jovem jornalista uma necessidade, esta evidência não dispensa o saber humanístico e a competência crítica, elementos que são diferenciadores do saber universitário e devem integrar o saber jornalístico. A imagem descarnada do produtor de conteúdos, um termo que parece extremamente redutor e uniformizador, desconsiderando as especificidades da profissão jornalística, é a manifestação de uma forma de unidimensionalidade da razão que se instala um pouco por toda a sociedade: a dependência exclusiva da aquisição da competência performativa cerca sobretudo aquelas profissões que se revelam mais frágeis na sua afirmação e, consequentemente, mais frágeis nos seus direitos e na sua autonomia. Uma segunda grande transformação faz-se sentir através de uma enorme pressão no sentido da concentração da propriedade. Nos nossos dias, a tendência inclui globalmente a constituição de grupos multimédia, nos quais as empresas procedem a investimentos simultâneos na televisão, rádio, imprensa e multimédia. A concentração da propriedade dos media pode induzir ameaças significativas ao pluralismo jornalístico; impor-se como um constrangimento à diversidade e como uma compulsão para a uniformização e redução de conteúdos; além de gerar fenómenos de auto-censura e uma maior sujeição dos jornalistas às directrizes empresariais, quer em matéria de conteúdo quer de políticas de gestão dos recursos humanos (Sousa, 2004, p. 2). Desde os anos 80 que se verifica a reentrada em cena dos grandes grupos económicos, gerando uma grande actividade financeira traduzida por fusões e aquisições e cruzamentos de acções. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso As novas alterações estruturais da indústria mediática conduziram ao grande aumento do custo de produção, devido à necessidade de reconversão tecnológica; necessidade de grandes investimentos na área do marketing, promoção e publicidade, segmentação da oferta e instauração de uma concorrência feroz (Correia, 1997, p. 70). Verificam-se, na área da informação, necessidades crescentes de integração multimedia em outros sectores das indústrias culturais para potenciar as possibilidades de promoção recíproca e induzir retornos rápidos do investimento. Estas alterações traduziram-se numa luta desenfreada pelas audiências com o aumento das soft news e também numa redução significativa de custos em detrimento dos recursos humanos destinados a outro tipo de informação, assim como na concentração de um número maior de tarefas num número menor de trabalhadores. A questão que se levanta na integração em grandes cadeias e conglomerados tem a ver com o efeito que a necessidade de retorno de investimento tem sobre o jornalismo praticado. Quanto mais lucro a firma exige, menos dinheiro disponível existirá para ser dispendido em jornalistas e cobertura noticiosa e mais delegações serão encerradas. Torna-se interessante verificar algumas das consequências que resultam dos fenómenos de abertura total ao mercado. Estes fenómenos conduzem a uma oferta padronizada e homogeneizada, que atende às necessidades de uma criação de produtos que satisfaçam expectativas e necessidades do maior número. Graças a esta tendência, informação, publicidade e entretenimento fundem-se num único objecto. O jornalismo é mais do que as empresas de jornalismo: é um olhar particular sobre o mundo que requer o estabelecimento de um contrato com os seus públicos. Logo, o sucesso do jornalismo terá que ser mais do que a eficácia das empresas e da criação de um produto, resultantes da afirmação unilateral de uma única dimensão da racionalidade, a racionalidade instrumental e finalista. Toda a problemática há muito levantada pela Teoria Crítica – com destaque para a Escola de Frankfurt – em torno da estandardização dos produtos culturais resultantes da penetração do pensamento administrativo no seio da cultura pode, com as devidas reservas devidas ao tempo e ao modo, serem relembradas. Uma terceira grande transformação verifica-se, pois, na própria natureza das mensagens liminarmente reduzidas a conteúdos da indústria, as quais conhecem uma tabloidização acentuada, uma fusão acentuada com o entretenimento e um aligeiramento de conteúdos. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Esta transformação estrutural da economia e da sociedade traduz-se num centramento cada vez mais insistente na aquisição de competências performativas em detrimento 6/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso (expresso na maioria dos editoriais, na escolha dos colunistas e comentadores externos, etc.) sobre os grandes temas da actualidade nacional e internacional e na menorização ou silenciamento das opiniões discordantes; na desvalorização das diferenças, dos grupos minoritários, dos trabalhadores e das suas organizações; (…) a concepção da informação como uma mercadoria que é preciso vender depressa e bem, leva ao predomínio não só do sensacional, mas também do curto, rápido e simples, do superficial e facilmente digerível, reflectindo a tendência para a supremacia dos valores comerciais sobre os valores jornalísticos. (Correia, s/d). Um dos problemas que urge ter em conta é a possibilidade de estas transformações gerarem movimentos de desprofissionalização e de proletarização do exercício do jornalismo. A desprofissionalização seria, segundo Marie Haug, (citada por Fidalgo, 2008: 53) resultaria do conjunto de factores como sejam a evolução tecnológica que torna mais acessível e difundido o conhecimento (já não só aos iniciados mas também aos leigos o que é manifestamente visível em movimentos como o jornalismo cidadão); a revalorização progressiva dos saberes empíricos, susceptíveis de serem aprendidos por pessoas sem formação académica; a crescente especialização dos domínios profissionais; a crítica crescente, por parte dos consumidores ao elitismo, à impunidade e à mercantilização de muitos profissionais e as reivindicações progressivas de clientes no sentido de uma maior participação nas decisões dos próprios profissionais. Numa relação verdadeiramente profissional, o trabalhador, porque possui o background teórico, dispõe do monopólio na formação do juízo e o cliente não pode fazer o diagnóstico nem propor a solução para o seu problema. Quando uma ocupação caminha na direcção da profissionalização, uma das suas aspiração é adquirir este monopólio. Entre os poderes associados ao exercício de formação está o exercício do controlo sobre os mecanismos de formação e instrução, isto é os mecanismos que se associam à aquisição das competências que permitem exercer o seu juízo e o seu parecer, aplicar o seu saber, em condições de monopólio (cf. Traquina, 2004: 46). O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Neste sentido, uma das propostas de reforma do jornalismo que recentemente tem adquirido adeptos – o Jornalismo do Cidadão – pode ser a expressão prática e intelectual de tendências que promovem a desprofissionalização ao insistir numa espécie de dicotomia entre os jornalistas e o publico que se configura como um confronto entre “nós” (os cidadãos sensibilizados para a comunicação livre e sem restrições) e “eles” (os jornalistas enquanto adversários ou obstáculos a essa liberdade, descritos como confinados a procedimentos rígidos e ciosos da sua posição de “privilégio” como detentores da informação) (cf. Moretzsohn, 2007: 264). Gillmor (2004) assume a existência de uma dicotomia entre a passividade do público no modelo tradicional do broadcast (one-to-many) , e o modelo de “thinking network” (many-to-many) característico dos blogs e outras manifestações próprias da Internet. Daqui surge o conceito de prossumidor, híbrido de produtor e consumidor. Assim, considera-se que , com a Internet e o jornalismo cidadão “ a venerável profissão de jornalista encontra-se num momento raro da sua história no qual, pela primeira vez, a sua hegemonia como gatekeeper de notícias está ameaçada não apenas por novas tecnologias e novos concorrentes mas, potencialmente, pela própria audiência a que serve” (Bowman e Willis, 2003: 7). Outro movimento seria a proletarização, um processo em que as condições de trabalho conduzem numerosas profissões à condição de assalariados, e que se verificaria nas próprias profissões estabelecidas exercidas em regime liberal (médicos, engenheiros, advogados). Esta proletarização implica o declínio de competências e qualificações dos profissionais e da sua autonomia de trabalho, graças a um processo em que pela fragmentação, desqualificação e rotinização das actividades profissionais, os trabalhadores tornar-se-iam cada vez mais meros executantes (cf. Fidalgo, 2008: 56). Estas tendências seriam particularmente visíveis no campo dos jornalistas, gerando debates há muito enraizados no seio desta profissão. A evolução tecnológica permitiu o surgimento de actividades parajornalísticas, ferindo o tradicional monopólio jornalístico, no tratamento, recolha e difusão de informação noticiosa sobre a actualidade. Simultaneamente generalizou-se as situações em que as fontes se dirigem directamente ao público, prescindindo da intermediação dos jornalistas e as situações em que o público acede , ele próprio directamente, às fontes (cf. Fidalgo, 2008: 60). A identificação destas transformações implica um olhar crítico em que se parte do pressuposto de que a forma como os jornalistas são formados não é um problema exclusivo dos jornalistas mas, antes, da sociedade em geral. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Ao longo desta linha de transformações há que reconhecer a existência de um importante eixo económico que constrange a qualidade da cidadania: “(…)A pobreza do debate e a fragilização da democracia revelam- se nas discriminações ideológicas, políticas, económicas, sociais, culturais, étnicas, geográficas e outras; no consenso 7/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso atitude problematizante. Antes se exigem, da parte de um certo sector identificado com específica visão do jornalismo que o reduz a sua natureza de indústria, jovens eficazes e pró-activos, convenientemente instigados pelo risco permanente da instabilidade laboral, conhecedores competentes de informática os quais prescindirão de um saber hermenêutico crítico, de uma capacidade de leitura da realidade para eficazmente a saberem «reproduzir». No limite, o jornalismo conformar-se-á com uma atitude natural e com o exercício de um profissionalismo anónimo, que se traduz na aplicação de rotinas pré- estabelecidas. Gaye Tuchman (1978) mostrou que os jornalistas usam esquemas classificatórios de forma a reduzir a contingência intrínseca ao trabalho noticioso, transformando ocorrências idiossincráticas verificadas no dia-a-dia em materiais que podem facilmente processados e divulgados. Assim, gera-se uma espécie de cristalização da experiência que assegura celeridade e eficácia ao desempenho profissional. Analisando a linguagem jornalística, detecta-se um conjunto de procedimentos, fórmulas discursivas, técnicas narrativas e descritivas que se repetem, parecendo, de certo modo, ser os acontecimentos que aderem a fórmulas narrativas pré-existentes e não o inverso. Um dos problemas desta mecanização da linguagem dos media é o ser facilmente associada a fenómenos como sejam a simplificação arbitrária dos acontecimentos; a generalização de estereótipos; o exercício de um constrangimento oculto muitas vezes associado ao funcionamento anónimo da organização e do grupo sobre a criatividade individual e a capacidade iniciativa dos jornalistas. A formação universitária dos jornalistas tem que incluir saberes que desenvolvam no ethos profissional do jornalista a capacidade de pensar reflexivamente sobre as suas práticas não por um culto estéril da observação mas pela necessidade de reinventar tais práticas, mudando-as de forma a que estas se tornem mais atentas às fenómenos periféricos, menos dependentes de enquadramentos rotineiros, mais sensível às vozes diversificadas de um universo cada vez mais fragmentado e plural. Neste sentido, os movimentos de reforma do jornalismo mais comprometidos com a deliberação democrática, como por exemplo o jornalismo cívico, desenvolvem metodologias que permitas redacções ficarem menos dependentes de uma hierarquia de definidores primários que enquadram ideologicamente o processo de construção social da realidade (Tanni Hass, 2007). Os jornalistas têm de se confrontar com pensamento novo sobre o jornalismo, para poderem re-avaliar e re-orientar práticas e experiências descobrindo formas de equacionar e representar a realidade. Este pensar crítico não é uma deriva especulativa estéril: recorre a competências específicas e a metodologias próprias. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Logo, a problemática do jornalismo não é apenas um problema que diz respeito ao jornalismo e aos jornalistas. Diz respeito a questões centrais da legitimidade democrática e da convivência cívico. Acreditamos, pois, que o jornalismo é crucial para a democracia, logo a forma como os jornalistas aprendem como exercer a profissão é um assunto que diz respeito à esfera pública. A complexidade crescente das sociedades exige outros saberes que permitam ultrapassar a "dignissíma tarimba", cuja exaltação se confunde aliás, com a ideia que de «o competente produtor de conteúdos» será apenas uma espécie de técnico multimédia nem sequer necessariamente demasiado culto – a cultura poderá tornar-se indutora de uma incómoda 8/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso sua relevância cívica. O ensino do jornalismo passará por uma alquimia de bom-senso entre a Academia e os diferentes sectores envolvidos no desempenho profissional: profissionais e organizações representativas e empresas. Dificilmente se obterá um consenso pois se suspeita que as discussões epistemológicas não são imunes às relações sociais e de poder. Todavia, suprimir um dos lados da equação, declarando-o irrelevante revela cegueira e incapacidade de compreender as facetas múltiplas da formação jornalísticas. Não atribuir relevo a formação académica e universitária no acesso à profissão significa regressar ao século XIX, antes de a autonomia do campo jornalístico determinar que este era uma profissão que não se reduz a um talento natural e ao autodidactismo. Neste sentido, o ensino universitário do jornalismo será também o ensino dirigido para uma leitura crítica da experiência. Esta é uma exigência social que transcende a problemática corporativa – a qual pelo contrário, por vez se ilude na reprodução prazenteiras das técnicas metamorfoseadas em rotinas – para se erguer ao estatuto de uma reclamação cívica. g p p g q p corporativa – a qual pelo contrário, por vez se ilude na reprodução prazenteiras das técnicas metamorfoseadas em rotinas – para se erguer ao estatuto de uma reclamação cívica. 1Professor da Universidade da Beira do Interior, Portugal. E-mail jcfcorreia@gmail.com For decades, journalism was seen as a simple practice (an occupation), which could exist without the need for specialized academic training. In this paper we defend the university as the most suitable way for the challenges and demands of the profession, remembering that it implies a critical distance in relation to the concrete conditions of professional practice. Thus it is argued that the journalistic ethos, based on a pragmatic attitude towards reality, will deal also with the critical requirements that characterize a knowledge theoretically oriented. This problem repeatedly taken up in various discussions (vocational versus academic knowledge, theory versus practice, reflective capabilities versus performative skills) is directly related with a reflection on the nature of the profession. BOWMAN, S. and WILLIS, C. 2003. We Media: how audiences are shaping the future of news and information. The Media Center at the American Institute, 65p. Acessado em: 03/03/2009, disponível em: http://www.hypergene.net/wemedia/download/we_media.pdf ANAVILHAS, J. 2009. O ensino do Jornalismo em Portugal. Estudos em Comunicação,6 (no prelo). CANAVILHAS, J. 2009. O ensino do Jornalismo em Portugal. Estudos em Comunicação,6 (no pre CORREIA, F. 1997. O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Lee Bollinger que presidiu à Task Force que analisou a reforma da Columbia Journalism School, defendeu que as escolas de jornalismo deverão saber manter a necessária «distância» das condições concretas de exercício da profissão, para poderem manter uma perspectiva independente em relação ao exercício do jornalismo. A lógica de uma formação integral continua a exigir que os novos profissionais tragam consigo um saber delimitado e específico mas igualmente universalista e crítica: em suma, um saber de Universidade, suficientemente crítico para não se esgotar na pura formação profissional, suficientemente ágil para não se reduzir ao academismo estéril e alheio à vida. Conclui-se assim que perante fenómenos complexos e imbricados entre si como sejam as transformações económicas, tecnológicas e de conteúdo, se erga uma necessidade de a aquisição de novos saberes teórico-metodológicos transponíveis, adaptáveis às novas circunstâncias, a novos contextos profissionais e organizacionais. Uma componente essencial da renovação dos saberes implica uma articulação cuidadosa entre a reflexão crítica e a aquisição de competências profissionais pelo que se torna pertinente equacionar a articulação entre disciplinas teóricas e disciplinas práticas (cf. Pinto, 2004: 59). Uma visão unilateral e determinista da formação assente na ausência de relacionamento entre dois mundos pode inibir o desenvolvimento de processos reflexivos e críticos ao nível das práticas e, ao mesmo tempo, simetricamente, tornar a reflexão prática, irrelevante, estéril e carente de sentido. Como se expressa provavelmente uma tal formação? Provavelmente, na co-existência de disciplinas crítico – conceptuais dirigidas para a análise e investigação sobre a área (Teoria da Notícia, Análise do Discurso da Imprensa, História da Imprensa), disciplinas prospectivas orientadas para a compreensão das dinâmicas em que se desenvolve o exercício das praticas jornalísticas (Jornalismo e Inovação ou Novos Jornalismos, duas designações para o mesmo conteúdo em Espanha e Portugal; Jornalismo Comparado, Jornalismos Especializados), disciplinas de orientação metodológica (quantitativa e qualitativa) e disciplinas de recorte laboratorial e experimental (Ateliers e Oficinas de Expressão, Técnicas de Redacção Jornalística, Géneros Jornalísticos etc.). Porém uma articulação deste jaez com as necessárias complementaridades e interacções indicia uma clara opção universitária. Tendencialmente, parece cada vez mais difícil pensar o ensino do Jornalismo sem a Universidade. Os passos atrás nesta opção parecem dificilmente sustentáveis do ponto de vista dos desempenhos da profissão, da sua legitimação social, da 9/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 10/02/2020 10/02/2020 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso Os jornalistas e as notícias, Lisboa, Editorial Caminho, 286 páginas. CORREIA, F (s/d), Jornalismo em Portugal. Acessado em: 21/10/2009, disponível em: http://resistir.info/portugal/fcorreia_jornalismo.html CORREIA, J. C. 1998. Algumas Reflexões sobre a Importância da Formação Universitária dos Jornalistas. BOCC. Acessado em: 22/10/2009, disponível em: http://www.bocc.ubi.pt/pag/correia-joao-formacao-universitaria-jornalistas.pdf pp 1-8 CORREIA, J. C. 1998. Algumas Reflexões sobre a Importância da Formação Universitária dos Jornalistas. BOCC. Acessado em: 22/10/2009, disponível em: http://www.bocc.ubi.pt/pag/correia-joao-formacao-universitaria-jornalistas.pdf pp 1-8 CORREIA, J.C. 2007. Apresentação. In: J. FERREIRA; E. VIZER. Mídia e Movimentos Sociais: Linguagens e colectivos em acção. São Paulo, Paulus, p. 9-19. CORREIA, J.C. 2007. Apresentação. In: J. FERREIRA; E. VIZER. Mídia e Movimentos Sociais: Linguagens e colectivos em acção. São Paulo, Paulus, p. 9-19. FRENCH, N. 2006. Competing Discourses of Journalism Education. In: IAMCR 25th Conference Proceedings. Cairo, p. 458- 465 GILLMOR, D. 2004. We the media. Grassroots Journalism by the People, for the People. Stanford, O’Relly, 320p AAS, T. 2007. The Pursuit of public journalism: theory, practice and criticism. Londres, Routledege, 254p. MARINHO, S. 2003. Reflexão sobre a necessidade de um novo paradigma para o ensino do Jornalismo: o caso da Universidade de Columbia. Acessado em: 21/10/2009, disponível em: http://repositorium.sdum.uminho.pt/bitstream/1822/6102/1/MarinhoS_universidadcolumbia_06.pdf MESQUITA, M. e PONTE, C. 1997. Situação do Ensino e da Formação Profissional na Área do Jornalismo. Acessado em: 21/10/2009, disponível em: http://www.labcom.ubi.pt/~bocc/pag/mesquita-mario-ponte-cristina-Cursos-Com3.html ORETZHON, S. 2007. Pensando contra os factos. Jornalismo e cotidiano: do senso comum ao sentido críti aneiro, Revan, 302 páginas. INTO, M. 2004. O ensino e a formação na área do jornalismo em Portugal: crise de crescimentoe notas pr omunicação e Sociedade, 5(1):49-62. 10/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 O ethos jornalístico: da técnica à reflexão crítica \| Correia \| Verso e Reverso 10/02/2020 10/02/2020 SOUSA, J.P. 2004. Desafios do ensino universitário do jornalismo ao nível da graduação no início do século XXI. Acessado em: 21/10/2009, disponível em http://bocc.ubi.pt/pag/_texto.php3?html2=sousa-jorge-pedro-desafios-do-jornalismo.html . TUCHMAN, G. 1978. Making News, New York, Free Press, 244 páginas. SOUSA, J.P. 2004. Desafios do ensino universitário do jornalismo ao nível da graduação no início do século XXI. Acessado em: 21/10/2009, disponível em http://bocc.ubi.pt/pag/_texto.php3?html2=sousa-jorge-pedro-desafios-do-jornalismo.html . ISSN: 1806-6925 - Melhor visualizado no Mozilla Firefox Este trabalho está licenciado sob uma Licença Creative Commons Attribution 3.0 São Leopoldo, RS. Av. Unisinos, 950. Bairro Cristo Rei, CEP: 93.022-000. Atendimento Unisinos +55 (51) 3591 1122 Projeto Gráfico: Jully Rodrigues ISSN: 1806-6925 - Melhor visualizado no Mozilla Firefox . Av. Unisinos, 950. Bairro Cristo Rei, CEP: 93.022-000. Atendimento Unisinos +55 (51) 3591 1122 p , , , ( ) Projeto Gráfico: Jully Rodrigues 11/11 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987 revistas.unisinos.br/index.php/versoereverso/article/view/5768/2987
https://openalex.org/W2921247115	https://researchonline.lshtm.ac.uk/id/eprint/4653306/1/the-ethical-imperative-to-treat-NCDs.pdf	English	null	The ethical imperative to treat NCDs during research in Africa	The Lancet. Global health/The lancet. Global health	2,019	cc-by	1,608	As the rising prevalence and life-threatening consequences of non-communicable diseases (NCDs) in Africa have become more obvious,1 so too has the fact that many African health systems are ill-equipped to meet the health-care needs of local communities affected by NCDs.2 Meanwhile, the quantity of clinical research done in Africa continues to increase,3 and carries with it unique ethical questions, especially relating to the amount of NCD medical care that ought to be provided for participants involved in this research. Many African research participants, whether enrolled in studies that investigate NCDs or other diseases, will inevitably have NCDs, but have little to no access to NCD health care in their communities. NCDs present a unique challenge because, contrary to acute infectious processes, the treatment of NCDs is not usually urgent and a lack of care will not lead to immediate morbidity or mortality. As such, do researchers in Africa have a responsibility to provide ancillary NCD services to research participants? In light of the third UN High- Level Meeting on NCDs, our aim is to provoke a dialogue regarding ancillary NCD services in the context of research in Africa. We believe that this dialogue can be guided by several lessons learned during the HIV epidemic and by a moral responsibility to provide ancillary care as articulated by Richardson.4 of ART being provided for participants in HIV trials.8,12,13 Members of one HIV trial network noted that, “One research organization cannot reverse global inequities in HIV[-1] care but researchers…have an obligation to try to narrow the equity gap.”12 of ART being provided for participants in HIV trials.8,12,13 Members of one HIV trial network noted that, “One research organization cannot reverse global inequities in HIV[-1] care but researchers…have an obligation to try to narrow the equity gap.”12 Third, study-specific guidelines regarding ancillary care for NCDs should be guided by the “four Ps”: positive duty, planning, partnership, and practical provisions.10 Researchers have a duty to assist study participants diagnosed with NCDs: careful planning is needed to anticipate the NCD burden in the study population; NCD care plans should be developed in partnership with the local community to prevent displacing or disrupting the local health-care system; and investigators should clearly state the planned NCD provisions. Comment Comment Comment Comment www.thelancet.com/lancetgh Vol 7 April 2019 Comment to help combat the NCD epidemic in Africa through the provision of appropriate ancillary services for NCDs. Further dialogue is needed to help researchers and sponsors articulate the extent of this moral imperative. 1 Naghavi M, Forouzanfar MH. Burden of non-communicable diseases in sub-Saharan Africa in 1990 and 2010: Global Burden of Diseases, Injuries, and Risk Factors Study 2010. Lancet 2013; 381: S95. 1 Naghavi M, Forouzanfar MH. Burden of non-communicable diseases in sub-Saharan Africa in 1990 and 2010: Global Burden of Diseases, Injuries, and Risk Factors Study 2010. Lancet 2013; 381: S95. WHO. A global brief on hypertension: silent killer, global public health crisis. World Health Day 2013. Geneva: World Health Organization, 2013. 3 World Bank. A decade of development in sub-Saharan African science, technology, engineering and mathematics research. Washington, DC: World Bank Group, 2014. Majaliwa Mzombwe, Bernard Desderius, Saidi Kapiga, Majaliwa Mzombwe, Bernard Desderius, Saidi Kapiga, Luke Smart, Robert Peck Vanderbilt University School of Medicine, Nashville, TN 37232, USA (MM); Center for Global Health and Department of Medicine, Division of General Internal Medicine, Weill Cornell Medical College, New York, NY, USA (RP); Department of Medicine, Weill Bugando School of Medicine, Mwanza, Tanzania (RP); Department of Medicine, Catholic University of Health and Allied Sciences, Mwanza, Tanzania (BD); London School of Hygiene and Tropical Medicine, Medical Research Council Tropical Epidemiology Group, London, UK (SK); Mwanza Intervention Trials Unit, National Institute for Medical Research, Mwanza, Tanzania (SK); and Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA (LS) j li b @ d bilt d *Majaliwa Mzombwe, Bernard Desderius, Saidi Kapiga, Luke Smart, Robert Peck Vanderbilt University School of Medicine, Nashville, TN 37232, USA (MM); Center for Global Health and Department of Medicine, Division of General Internal Medicine, Weill Cornell Medical College, New York, NY, USA (RP); Department of Medicine, Weill Bugando School of Medicine, Mwanza, Tanzania (RP); Department of Medicine, Catholic University of Health and Allied Sciences, Mwanza, Tanzania (BD); London School of Hygiene and Tropical Medicine, Medical Research Council Tropical Epidemiology Group, London, UK (SK); Mwanza Intervention Trials Unit, National Institute for Medical Research, Mwanza, Tanzania (SK); and Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA (LS) majaliwa mzombwe@vanderbilt edu 4 Richardson HS. Moral entanglements: the ancillary-care obligations of medical researchers. Oxford: Oxford University Press, 2012. 5 Benatar SR, Singer PA. A new look at international research ethics. BMJ 2000; 321: 824–26. 6 Dickert N, Wendler D. Fourth, study-specific ancillary care plans for NCDs should be guided by the nature of the relationships between the study participants, the investigators, and the community.4 One working group suggested a series of questions helpful in evaluating ancillary care needs, including “how severe or acute are the likely ancillary care needs”, is the identification of NCDs “integral or incidental to carrying out study procedures”, and “how extensive are the proposed interactions between researchers and study participants”.10 The final plan for ancillary NCD services should be the result of a moral negotiation between community members and the research study team.11 First, researchers in Africa should strive for the “highest achievable standard” of ancillary care for NCDs diagnosed during the course of medical research.5 Setting standards that are too high for ancillary NCD services will make the conduct of research too burdensome.6,7 By contrast, accepting health inequity as an unchangeable reality misses the opportunity for research to combat these inequities.5,8–11 Determining this “highest achievable standard” will require an in- depth understanding of the social, economic, and political contexts of the settings for each study.5 Finally, in our own research14,15 in east Africa, we have learned that high-quality ancillary services for common NCDs can be provided through the local health system at a reasonable cost. For example, in a cohort study of adults with HIV funded by the US National Institutes of Health, we raised modest private funds to provide free treatment for hypertension and diabetes. The HIV clinic used these funds to purchase high-quality generic drugs from a local pharmacy. Furthermore, we helped the HIV clinic’s case manager to develop a system to track blood pressure, blood glucose control, and prescription refills. We also worked with HIV staff to incorporate hypertension and diabetes screening into routine clinic services and to develop locally relevant NCD health education. Second, the “highest achievable standard” should be continuously reassessed and increased on the basis of improvements in the local health-care system and new scientific knowledge. During the initial decades (1995–2010) of HIV research in Africa, for example, the local capacity to provide antiretroviral treatment (ART) increased dramatically.12 Therefore, the standard for ancillary HIV care in Africa also increased to the point In conclusion, the medical research community in Africa has the opportunity and the moral responsibility e406 Comment www.thelancet.com/lancetgh Vol 7 April 2019 Comment Ancillary care obligations of medical researchers. JAMA 2009; 302: 424–28. 7 Dickert N, DeRiemer K, Duffy PE, et al. Ancillary-care responsibilities in observational research: two cases, two issues. Lancet 2007; 369: 874–77. 8 8 Council for International Organizations of Medical Sciences. International Ethical Guidelines for Health-related Research Involving Humans. Geneva: Council for International Organizations of Medical Sciences, 2016. Council for International Organizations of Medical Sciences, 2016. 9 Lurie P, Sidney M. Unethical trials of interventions to reduce perinatal transmission of the human immunodeficiency virus in developing countries. N Engl J Med 2000; 337: 853–56. 10 Brownsword R, Cermak A, Chaisson R, et al. The ancillary-care obligations of medical researchers working in developing countries. PLoS Med 2008; 5: 709–13. 11 Slack CM. Ancillary care in South African HIV vaccine trials: addressing needs, drafting protocols, and engaging community. J Empir Res Hum Res Ethics 2014; 9: 83–95. majaliwa.mzombwe@vanderbilt.edu This work was supported by the National Institutes of Health Office of the Director, Fogarty International Center, Office of AIDS Research, National Cancer Center, National Eye Institute, National Heart, Blood, and Lung Institute, National Institute of Dental and Craniofacial Research, National Institute On Drug Abuse, National Institute of Mental Health, National Institute of Allergy and Infectious Diseases, and National Institutes of Health Office of Women’s Health and Research through the Fogarty International Clinical Research Scholars and Fellows Program at Vanderbilt University (R24 TW007988) and the American Recovery and Reinvestment Act. All authors declare no competing interests. We would like to thank Ellen Clayton and Doug Heimburger. 12 Fitzgerald DW, Pape JW, Wasserheit JN, Counts GW, Corey L. Provision of treatment in HIV-1 vaccine trials in developing countries. Lancet 2003; 362: 993–94. 13 UNAIDS. Ethical considerations in HIV preventive vaccine research UNAIDS guidance document. Geneva: UNAIDS, 2000. 14 Peck RN, Shedafa R, Kalluvya S, et al. Hypertension, kidney disease, HIV and antiretroviral therapy among Tanzanian adults: a cross-sectional study. BMC Med 2014; 12: 125. 15 Kayange NM, Smart LR, Downs JA, Maskini M, Fitzgerald DW, Peck RN. The influence of HIV and schistosomiasis on renal function: a cross-sectional study among children at a hospital in Tanzania. PLoS Negl Trop Dis 2015; 9: e0003472. 15 Kayange NM, Smart LR, Downs JA, Maskini M, Fitzgerald DW, Peck RN. The influence of HIV and schistosomiasis on renal function: a cross-sectional study among children at a hospital in Tanzania. PLoS Negl Trop Dis 2015; 9: e0003472. Copyright © 2019 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. Copyright © 2019 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. e407 www.thelancet.com/lancetgh Vol 7 April 2019
https://openalex.org/W2961123289	https://periodicals.karazin.ua/economy/article/download/12585/11976	Ukrainian	null	Macroeconimic dynamic model with environmental factors (on the case of Ukraine)	Vìsnik Harkìvsʹkogo nacìonalʹnogo unìversitetu ìmenì V.N. Karazìna. Serìâ Ekonomìka	2,018	cc-by	3,325	Серія «Економічна», випуск 95, 2018 Серія «Економічна», випуск 95, 2018 DOI: 10.26565/2311-2379-2018-95-06 Т.В. Меркулова, О.В. Томілович  Харківський національний університет імені В.Н. Каразіна пл. Свободи, 4, м. Харків, 61022, Україна mara.merkulova@karazin.ua, ORCID: https://orcid.org/0000-0002- Т.В. Меркулова, О.В. Томілович  Харківський національний університет імені В.Н. Каразіна пл. Свободи, 4, м. Харків, 61022, Україна E-mail: tamara.merkulova@karazin.ua, ORCID: https://orcid.org/0000-0002-3507-5593 МОДЕЛЮВАННЯ, ІМІТАЦІЯ ТА ІНФОРМАЦІЙНІ ТЕХНОЛОГІЇ В ЕКОНОМІЦІ Й УПРАВЛІННІ УДК 330.3: 330.4: 338.2 МАКРОЕКОНОМІЧНА ДИНАМІЧНА МОДЕЛЬ З ВРАХУВАННЯМ ЕКОЛОГІЧНИХ ФАКТОРІВ (НА ПРИКЛАДІ УКРАЇНИ) Загострення екологічних проблем, які пов’язані з економічною діяльністю людини, поширення цього явища на всі (без винятку) регіони та державу світу, зумовлює актуальність і глобальність цього питання в наш час, показує необхідність проведень досліджень щодо вивчення взаємозв’язків між економічним розвитком і природними змінами для пошуку шляхів вирішення протирічь між ними. Важливу роль в таких дослідженнях відіграють економіко-математичні моделі, що дозволяють кількісно оцінити ці взаємозв’язки, проводити багатоваріантні розрахунки з метою пошуку ефективних варіантів економічного розвитку з урахуванням екологічної складової. ур у Метою даної роботи є розробка макроекономічної динамічної моделі з урахуванням екологічних факторів на підставі модифікації динамічної інтегрованої моделі економіки та клімату Вільяма Нордгауза (модель DICE) та її апробація на основі даних по Україні. У статті розглянуті наступні результати дослідження: проведений огляд основних припущень моделі Нордгауза; розроблено макроекономічну динамічну модель з врахуванням екологічних факторів за рахунок модифікації моделі Нордгауза; проведено оцінку параметрів модифікованої моделі на основі даних по Україні; створено системно-динамічну реалізацію цієї моделі; проведено експерименти з системно-динамічною реалізацією моделі; здійснено інтерпретацію результатів експериментів. р р р р у р Дослідження було проведено за допомогою методів математичного та системно-динамічного моделювання, статистичних методів, економетричного моделювання, з використанням програм Vensim та Microsoft Excel (зокрема інструменту Regression пакету Data Analysis). За результатами експериментів були надані кількісні оцінки певним взаємозв’язкам між економічним розвитком та деякими змінами природи і обґрунтовано висновок щодо можливості використання модифікованої моделі для аналізу та узгодження економічної та екологічної складової. д ф д д у у д д Ключові слова: модель Нордгауза, екологічні фактори розвитку, навколишнє середовище, сталий розвиток. JEL Cl ifi i Q 5 O 13 C 5 C 6 JEL Classification: Q 5, O 13, C 5, C 6. Tamara Merkulova, Oleksiy Tomilovych V.N. Karazin Kharkiv National University 4 Svobody sq., 61022, Kharkiv, Ukraine a.merkulova@karazin.ua, ORCID: https://orcid.org/0000-0002-3507-5593 ВІСНИК ХАРКІВСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ імені В. Н. КАРАЗІНА The objective of this study is to develop the macroeconomic dynamic model with environmental factors by modifying William Nordhaus Dynamic Integrated Climate-Economy Model (DICE model) and test it on the data of Ukraine. This article contains the following research results: the main assumptions of the Nordhaus model were outlined; the macroeconomic dynamic model with environmental factors was developed by modifying the Nordhaus model; the parameters of the modified model were estimated on the data of Ukraine; the system-dynamic implementation of the model was developed; experiments with the system-dynamic implementation of the model were conducted; the results of the experiments were interpreted. p p p The research was conducted with methods of mathematical and system-dynamic modeling, statistical methods, econometric modeling, Vensim and Microsoft Excel programs (the Data Analysis Regression tool in particular). p Based on the above, some patterns between economic development and nature changes were found, which confirmed the possibility of using the modified model for analyzing and reconciling these components. Key words: Nordhaus model, environmental factors of development, environment, sustainable development. C Q O C C p JEL Classification: Q 5, O 13, C 5, C 6. p JEL Classification: Q 5, O 13, C 5, C 6. Т.В. Меркулова, А.В. Томилович Харьковский национальный университет имени В.Н. Каразина пл. Свободы, 4, г. Харьков, 61022, Украина E-mail: tamara.merkulova@karazin.ua, ORCID: https://orcid.org/0000-0002-3507-5593 MACROECONIMIC DYNAMIC MODEL WITH ENVIRONMENTAL FACTORS (ON THE CASE OF UKRAINE) The aggravation of environmental problems, related to human economic activity, and the spread of the phenomenon to all (without exception) regions and states of the world determinate relevance and global dimension of this issue in these times and show the need for research on the relationship between economic development and nature change to find ways to resolve their contradiction. Economic mathematical models play a significant role in such research as they allow quantitative estimation of these interconnections and multivariate calculations in order to find effective alternatives for economic development with an environmental dimension.  Меркулова Т. В., Томілович О. В., 2018 49 МАКРОЭКОНОМИЧЕСКАЯ ДИНАМИЧЕСКАЯ МОДЕЛЬ С УЧЕТОМ ЭКОЛОГИЧЕСКИХ ФАКТОРОВ (НА ПРИМЕРЕ УКРАИНЫ) Обострение экологических проблем, которые связанны с экономической деятельностью человека, распространение этого явления на все (без исключения) регионы и государства мира, обусловливает актуальность и глобальность этого вопроса в наше время, показывает необходимость проведения исследований касаемо взаимосвязей между экономическим развитием и природными изменениями для поиска путей разрешения противоречий между ними. Важную роль в таких исследованиях играют экономико-математические модели, что позволяют количественно оценить эти взаимосвязи, проводить многовариантные расчеты с целью поиска эффективных вариантов экономического развития с учетом экологической составляющей. щ Целью данной работы есть разработка макроэкономической динамической модели с учетом экологических фактор на основании модификации динамической интегрированной модели экономики и климата Уильяма Нордхауса (модель DICE) и ее апробация на основании данных по Украине. В статье рассмотрены следующие результаты исследования: проведен осмотр основных допущений модели Нордхауса; разработано макроэкономическую динамическую модель с учетом экологических факторов за счет модификации модели Нордхауса; проведено оценку параметров модифицированной модели на основании данных по Украине; создано системно-динамическую реализацию этой модели; проведено эксперименты с системно-динамической реализацией модели; осуществлено интерпретацию результатов эксперимента. Исследование было проведено с помощью методов математического и системно-динамического моделирования, статистических методов, эконометрического моделирования, с использованием программ Vensim и Microsoft Excel (в частности инструмента Регрессия пакета Анализ данных). По результатам экспериментов были предоставлены количественные оценки определенным взаимосвязям между экономическим развитием и некоторыми изменениями природы и обоснован вывод о возможности использования модифицированной модели для анализа и согласования экономической и экологической составляющей. Ключевые слова: модель Нордхауса, экологические факторы развития, окружающая среда, устойчивое развитие. р JEL Classification: Q 5, O 13, C 5, C 6. JEL Classification: Q 5, O 13, C 5, C 6. Серія «Економічна», випуск 95, 2018 Серія «Економічна», випуск 95, 2018 змін клімату на певну галузь, зазвичай сільське господарство (Нечипоренко, 2016), як найуразливіше до цих процесів. І тільки в останні роки з’явилися роботи, що більш-менш різносторонньо розглядають взаємозв’язки між природою та економікою в масштабі України (Двуліт, 2015), (Горобець, 2017). (Д у ) ( р ) Початком системних закордонних досліджень цієї проблематики прийнято вважати ідеї Римського клубу (Club of Rome) та концепцію сталого розвитку (Meadows & al., 1972). Саме з них зміни природи почали розглядатися, з одного боку, як наслідок економічного розвитку і, з іншого, як його фактор. З самого початку таким взаємозв’язкам прагнули дати кількісну оцінку. Якщо спочатку такі оцінки носили експертний характер (так, Тол вказує, що ще в 1979 р. д’Арж оцінив відсоток втрат економіки від глобального потепління (Tol, 2018)), то згодом на зміну їм прийшли економіко-математичні моделі, які могли не тільки описувати взаємозв’язки, але й давати прогнози щодо них (Tol, 2018), (Nordhaus, 2013). На сьогоднішній день, як кількісна характеристика взаємовпливу економіки та природи, використовується показник “the social cost of carbon (SCC)” (Nordhaus, 2013&2016), (Tol, 2018), (Auffhammer, 2018), що може бути перекладено як суспільна вартість вуглецю — приблизні економічні втрати в майбутньому від випуску 1 т діоксиду вуглецю сьогодні. Ця характеристика використовується, зокрема, й для встановлення податків на викиди, дозволяючи в певній мірі регулювати їх. р у В основі цієї роботи лежить динамічна інтегрована модель клімату та економіка В. Нордгауза, в якій він включив кліматичні зміни до довгострокового економічного аналізу, за що отримав Нобелівську премію з економіки 2018 р. У статті представлено авторську модифікацію цієї моделі та результати її апробацію на основі даних по Україні. В основі цієї роботи лежить динамічна інтегрована модель клімату та економіка В. Нордгауза, в якій він включив кліматичні зміни до довгострокового економічного аналізу, за що отримав Нобелівську премію з економіки 2018 р. У статті представлено авторську модифікацію цієї моделі та результати її апробацію на основі даних по Україні. Актуальність дослідження та аналіз досліджень та публікацій Актуальність дослідження та аналіз досліджень та публікацій Дослідження впливу економічної діяльності людини на природу, навколишнє середовище, клімат стало особливо актуальним з початку XXI ст. Це можна пояснити загостренням екологічних проблем по всьому світу, тобто негативною зворотною реакцією природи на діяльність людини — “помстою природи”. р р В Україні такі дослідження носять здебільшого теоретичний характер (Зварич, 2016), основні постулати якого запозичені з іноземних робіт, або розглядають односторонній вплив 50 Серія «Економічна», випуск 95, 2018 Модифікована макроекономічна модель Модифікована макроекономічна модель В основі оригінальної моделі Нордгауза лежать наступні припущення: В основі оригінальної моделі Нордгауза лежать наступні припущення:  для споживання та заощадження використовується чистий продукт — валовий продукт за вирахуванням втрат від зміни клімату та витрат на скорочення викидів;  в процесі виробництва відбуваються викиди діоксиду вуглецю, пропорційні валовому продукту;  збільшення викидів діоксиду вуглецю опосередковано призводить до підвищення температури повітря; р ур р  підвищення температури призводить до збільшення втрат від зміни клімату;  витрати на скорочення викидів пропорційні валовому випуску;  коефіцієнти пропорційності викидів та витрат валовому продукту змінюються з часом, відображаючи вплив науково-технічного розвитку. дображаючи вплив науково-технічного розвитку. Відповідно до основних положень моделі Нордгауза, було розроблено модифіковану макроекономічну динамічну модель з врахуванням екологічних факторів з наступними рівняннями. Відповідно до основних положень моделі Нордгауза, було розроблено модифіковану макроекономічну динамічну модель з врахуванням екологічних факторів з наступними рівняннями. р Величина кінцевого продукту скорочується на величину витрат, пов’язаних з кліматом та екологією Величина кінцевого продукту скорочується на величину витрат, пов’язаних з кліматом та екологією Q(t)=Y(t)-TC(t), Q(t) — чистий кінцевий продукт, Y(t) — валовий внутрішній продукт, TC(t) — витрати на охорону атмосферного повітря та проблеми зміни клімату. Q(t) — чистий кінцевий продукт, Y(t) — валовий внутрішній продукт, TC(t) — витрати н охорону атмосферного повітря та проблеми зміни клімату. р у ф р р р у Саме чистий кінцевий продукт Q(t) використовується для споживання та заощадження Q(t)=C(t)+I(t), Саме чистий кінцевий продукт Q(t) використовується для споживання та заощадження Q(t)=C(t)+I(t), ( ) ( ) ( ), C(t)=(1-s)Q(t) — кінцеве споживання, I(t)=sQ(t) — капітальні інвестиції, 1-s і s — фіксован норма споживання та заощадження відповідно. ( ) ( ) ( ) C(t)=(1-s)Q(t) — кінцеве споживання, I(t)=sQ(t) — капітальні інвестиції, 1-s і s — фіксована норма споживання та заощадження відповідно. Валовий внутрішній продукт Y(t) моделюється за виробничою функцією Кобба-Дугласа Y(t)=AK(t)αL(t)1-α, Валовий внутрішній продукт Y(t) моделюється за виробничою функцією Кобба-Дугласа Y(t)=AK(t)αL(t)1-α, ( ) ( ) ( ) , A — технологічний коефіцієнт, K(t) — вартість основних засобів, L(t) — чисельність зайнятого населення, α і 1-α — коефіцієнт еластичності випуску по капіталу та трудових ресурсах відповідно. ( ) ( ) ( ) A — технологічний коефіцієнт, K(t) — вартість основних засобів, L(t) — чисельність зайнятого населення, α і 1-α — коефіцієнт еластичності випуску по капіталу та трудових ресурсах відповідно. змінної L(t) описується експоненціальним законом Динаміка змінної L(t) описується експоненціальним законом L(t) L nt Динаміка змінної L(t) описується експоненціальним законом L(t)=L0ent. Серія «Економічна», випуск 95, 2018 Зміна величини капіталу K(t) відбувається з урахуванням інвестицій та амортизації K(t+1)=K(t)+I(t)-D(t), Зміна величини капіталу K(t) відбувається з урахуванням інвестицій та амортизації K(t+1)=K(t)+I(t)-D(t), Зміна величини капіталу K(t) відбувається з урахуванням інвестицій та амортизації K( 1) K( ) I( ) D( ) 51 У свою чергу, викиди впливають на середню температуру повітря T̅ ( ) ϕ ϕ E̅ ( ) викиди впливають на середню температуру повітр ̅ ̅ р р у T̅ (t0, t)=ϕ0+ϕ1E̅ (t0, t), T̅ (t0, t) — середня температура повітря за період з t0 по t, E̅ (t0, t) — середні викиди за період з t0 по t, ϕ0 і ϕ1 — параметри. р ϕ ϕ р р Звідси середня температура T(t) періоду t визначається за формулою ( ) у T(t)=(t-t0+1)T̅ (t0, t)-(t-t0)T̅ (t0, t-1), Витрати на охорону атмосферного повітря і проблеми зміни клімату залежать від величини ВВП Y(t), температури T(t) і технології C( ) θ( ) ( ) ( ) ( ) ( ) ( ) ( ) θ(t) — технологічний параметр, що відображає скорочення витрат на одиницю ВВПградус Цельсія вище нуля з часом. Він також змінюється за експоненціальним законом. θ(t)=θ0eθ₁t. θ(t) — технологічний параметр, що відображає скорочення витрат на одиницю ВВПградус Цельсія вище нуля з часом. Він також змінюється за експоненціальним законом. θ(t)=θ0eθ₁t. ВІСНИК ХАРКІВСЬКОГО НАЦІОНАЛЬНОГО УНІВЕРСИТЕТУ імені В. Н. КАРАЗІНА D(t)=δKK(t) — величина амортизації, δK — фіксована норма амортизації. D(t)=δKK(t) — величина амортизації, δK — фіксована норма амортизації. У процесі економічної діяльності відбуваються викиди діоксиду вуглецю, пропорційні валовому внутрішньому продукту Y(t) ( ) ( ) р ф р р У процесі економічної діяльності відбуваються викиди діоксиду вуглецю, пропорційні валовому внутрішньому продукту Y(t) E(t)=σ(t)Y(t), ( ) ( ) ( ), E(t) — викиди діоксиду вуглецю стаціонарними джерелами, σ(t) — технологічний параметр, що відображає трендовий характер зменшення викидів на одиницю ВВП. Він змінюється аналогічно до трудових ресурсів σ(t)=σ0eσ₁t. σ(t)=σ0eσ₁t. ( ) У свою чергу, викиди впливають на середню температуру повітря ̅ ̅ ( ) У свою чергу, викиди впливають на середню температуру повітря ивають на середню температуру повітря ( Оцінка параметрів та апробація моделі Оцінка параметрів рівнянь модифікованої моделі була проведена на основі щорічних статистичних показників України з 2000 р., скоригованих з урахуванням впливу тимчасово окупованих територій Автономної Республіки Крим, м. Севастополя, частини Донецької та Луганської обл. (Держстат України). Вартісні показники при цьому були приведені до порівняних цін 2010 р., а нелінійні рівняння були попередньо лінериазовані. Під час параметризації використовувався інструмент Regression пакету Data Analysis програми Microsoft Excel. Отримані моделі виявилися адекватними за F-критерієм Фішера, а оцінки параметрів — значущими за t-статистикою Стьюдента з довірчою ймовірністю 95%. Звичайні й скориговані коефіцієнти детермінації, середні помилки апроксимації при цьому коливалися від допустимих до відмінних значень. Відповідно до отриманих оцінок чисельність зайнятого населення України скорочується на 1% щорічно. Викиди на одиницю ВВП і витрати на одиницю ВВП*градус Цельсія при цьому зменшуються на 2,3 і 6,1% відповідно. Також середня багаторічна температура підіймається на 0 015 градуси Цельсія при зростанні середніх щорічних викидів діоксиду вуглецю на 1 млн т На основі рівняння модифікованої моделі й оцінок їхніх параметрів було побудовано її системно-динамічну реалізацію за допомогою програми Vensim (рис.1). Рис. 1. – Структура системно-динамічної моделі в програмі Vensim Джерело: авторська розробка Рис. 1. – Структура системно-динамічної моделі в програмі Vensim Рис. 1. – Структура системно-динамічної моделі в програмі Vensim Джерело: авторська розробка 52 р ур Висновки Висновки На підставі експериментів були отримані наступні результати. На підставі експериментів були отримані наступні результати. ві експериментів були отримані наступні результат р у р у р у Зміні параметра α відповідає зміна динаміки ВВП: при менших значення спостерігається швидше падіння, при більших — швидше зростання, проте темп зміни ВВП збігається до темпу зміни чисельності зайнятого населення. При зміні норми заощадження s спостерігається зміна динаміки загального та подушного споживання: більшим значенням параметра відповідає програш на початку, який компенсується кращим ефектом в довгостроковій перспективі та навпаки. у р ф р р З іншого боку, було виявлено, що темп зміни викидів збігається до значення en+σ₁, якщо n+σ1≤1. Також, за умови падіння або стабілізації рівня викидів, темп зміни витрат сходиться до значення en+θ₁, а подушних викидів і витрат — eσ₁ і eθ₁ відповідно. У цілому дослідження з побудована модель зберігає основні закономірності макроекономічних динамічних моделей, заснованих на неокласичній виробничої функції. Великим значенням параметра викидів σ1 та витрат θ1 відповідає падіння екологічних характеристик виробництва, що негативно впливає на навколишнє середовище, призводить до збільшення витрат і зменшення споживання. У цілому дослідження з побудована модель зберігає основні закономірності макроекономічних динамічних моделей, заснованих на неокласичній виробничої функції. Великим значенням параметра викидів σ1 та витрат θ1 відповідає падіння екологічних характеристик виробництва, що негативно впливає на навколишнє середовище, призводить до збільшення витрат і зменшення споживання. Розроблені моделі можуть бути використані як інструмент якісного та кількісного аналізу взаємозв’язків між економічними та екологічними факторами для пошуку ефективних шляхів узгодження їх впливу на добробут населення. Серія «Економічна», випуск 95, 2018 Змінні “… попереднього періоду” цієї моделі задані на рівні 2016 р., змінні-рівні — 2017 р. На її основі були проведені експерименти: багаторазові прогоні моделі на протязі 20 тактів для різних значень деякого параметра при початковому рівні інших заради виявлення чутливості деяких змінних, визначених як контрольні. Такими змінними були обрані ВВП, споживання, викиди та витрати, а також їхні подушні аналоги. Досліджуваними параметрами, в свою чергу, виступили: коефіцієнт еластичності випуску по капіталу α, норма заощадження s, норма амортизації dK, темп приросту населення n, параметр викидів σ0, параметр витрат θ0, температурний коефіцієнт ϕ0. р ур Висновки Література р ур 1. Зварич І. Сучасні глобальні екологічні ризики. Вісник Тернопільського національного економічного університету. 2016. Вип. 4. С. 95–101. у р у 2. Нечипоренко О. Стан та перспективи адаптації аграрного сектору економіки України до глобальних змін клімату. Економіст. 2016. №11. С. 10–14. 3. Двуліт З. 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https://openalex.org/W4390445774	https://www.ozguryayinlari.com/site/catalog/download/401/1950/4482	Turkish	null	Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik Büyüme: BRICS ve MINT Ülkeleri Üzerine Ampirik Bir İnceleme	Özgür Yayınları eBooks	2,023	cc-by	119,780	Editör: Published by Özgür Yayın-Dağıtım Co. Ltd. Certificate Number: 45503 15 Temmuz Mah. 148136. Sk. No: 9 Şehitkamil/Gaziantep +90.850 260 09 97 +90.532 289 82 15 www.ozguryayınlari.com info@ozguryayinlari.com Sosyal Bilimlerde Akademik Araştırma ve Değerlendirmeler-V Academic Research and Evaluations in Social Sciences-V Editör: Prof. Dr. Ali Şahin Sosyal Bilimlerde Akademik Araştırma ve Değerlendirmeler-V Academic Research and Evaluations in Social Sciences-V Editör: Prof. Dr. Ali Şahin Language: Turkish-English Publication Date: 2023 Cover design by Mehmet Çakır Cover design and image licensed under CC BY-NC 4.0 Print and digital versions typeset by Çizgi Medya Co. Ltd. ISBN (PDF): 978-975-447-850-1 ISBN (PDF): 978-975-447-850-1 DOI: https://doi.org/10.58830/ozgur.pub401 This work is licensed under the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0). To view a copy of this license, visit https://creativecommons.org/licenses/by-nc/4.0/ This license allows for copying any part of the work for personal use, not commercial use, providing author attribution is clearly stated. Suggested citation: gg Şahin, A. (ed) (2023). Sosyal Bilimlerde Akademik Araştırma ve Değerlendirmeler-V. Özgür Publications. DOI: https://doi.org/10.58830/ozgur.pub401. License: CC-BY-NC 4.0 The full text of this book has been peer-reviewed to ensure high academic standards. For full review policies, see https://www.ozguryayinlari.com/ Önsöz Önsöz Teknolojik gelişmeler ve küresel anlamda gerçekleşen değişimler insan doğasını değiştirmektedir. Bu değişim insan hayatının tüm aşamalarına doğrudan etki etmektedir. Bu noktada insana ve insan yaşantısına yönelik olarak araştırmalar gerçekleştirmek kaçınılmaz bir hal almaktadır. Sosyal Bilimler alanında gerçekleştirilen tüm çalışmalar insanı, insan yaşantısını ve insan doğasını anlamayı amaçlamaktadır. İnsanı anlamak adına ortaya koyulan bu eserde farklı disiplinlerden farklı çalışmalar ele alınmış ve bir nebze de olsa bu amaca ulaşmak için bir basamak inşa edilmeye çalışılmıştır. Preface Technological developments and globalization has changed human nature. This change directly affects all stages of human life. At this point, it becomes inevitable to carry out research on people and human life. All studies carried out in the field of Social Sciences aim to understand human, human life and human nature. In this work, which was put forward in order to understand the human, different studies from different disciplines were discussed and a step was tried to be built in order to reach this goal, albeit to some extent. iii iii İçindekiler iii iii iii iii Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 1 Orhan Ecemis Asli Ozpolat Ferda Nakipoglu Ozsoy 1 1 Orhan Ecemis Asli Ozpolat Ferda Nakipoglu Ozsoy Türkiye’de Gelir Dağılımına İlişkin Görünüm Bengü Kayapalı Aslan 23 Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 71 71 Bengü Kayapalı Aslan Bengü Kayapalı Aslan Bölüm 6 Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin ğ j Ekonomik Büyüme Üzerindeki Etkisine İlişkin Bir İnceleme: Türkiye Örneği 85 Elif Koçak 97 Bölüm 3 Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik Büyüme: BRICS ve MINT Ülkeleri Üzerine Ampirik Bir İnceleme 35 Şahin Nas Ömer Demir Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL 53 Sacit Sarı v mı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Mehmet Kaya Muhammed Çelik 125 179 vi Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi 197 Fazilet Nohut Ö Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi 197 Fazilet Nohut Özlem Balaban Fazilet Nohut Özlem Balaban Bölüm 13 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 221 Ünal Arslan Metin Kılıç Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik Eğitimi 243 Şahin Daştan Mehmet Göl Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 271 Serhat Ata Ali Özer Nevin Özer 271 303 Kâr Payı Dağıtımı: Politika ve Teoriler Emine Kaya Ömer Kaya 315 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri vii vii Bölüm 18 1 Department of Computer Technology, Gaziantep University, Türkiye, oecemis@gantep.edu.tr 2 Department of Management and Organization, Gaziantep University, Türkiye, ozpolat@gantep.edu.tr 3 Department of Economics, Gaziantep University, Türkiye, nakipoglu@gantep.edu.tr, Abstract Sustainable Development Goal significantly indicates the clean energy, climate actions and environmental development. Especially with the European Green Deal’s coverage of international trade, many countries have accelerated their efforts to reduce their emission rates. In addition, the fact that the importance to be taken differs on a sectoral basis, which requires the separation of emission rates by sector and the necessary measures to be taken. This study discusses the possible consequences of including Turkey in the emissions trading system within the scope of the green deal and Sustainable Development Goal (SDG 13 and SDG 15) in the light of the reasons stated. For this prediction, the m5p method, one of the data mining decision tree methods, was used. The model examined. According to the data obtained, environmental taxes currently do not have any effect on emission the effects of environmental taxes on CO2 emissions and estimated the emission value for the next 5 years if taxes were increased values in Turkey. Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models Orhan Ecemis1 Asli Ozpolat2 Ferda Nakipoglu Ozsoy3 Chapter 1 Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models Chapter 1 Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar Berna Ağbulut Bölüm 19 Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı ve Kârlılık Göstergeleri Açısından Analizi 343 Mehtab Çelik Ersin Korkmaz Mehtab Çelik Ersin Korkmaz Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi ile Değerlendirilmesi 361 Aslı Uğur Aydın Taştekin Ayşegül Güdül Posta, Haberleşme ve Kargo İ ile Değerlendirilmesi Aslı Uğur Aydın Taştekin Ayşegül Güdül Kollektif Şirkette Ortaklığa Giriş Yunus Alhan 375 viii 1. Introduction When we look at the development process of the EU climate policy, the first attempts were made in the 1980s and 1990s, and it started to be developed step by step in the 2000s and 2010s. In 2019, important steps have been taken with the European Green Deal (Delbeke & Vis, 2019; Kulovesi https://doi.org/10.58830/ozgur.pub401.c1749 1 1 2 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 2 & Oberthür, 2020). At this point, it is known that the EU has an important role. It is an accepted fact that the EU has the role of an international climate leader, as it creates the most advanced climate policy framework among the world’s leading actors (Jordan & Moore, 2020; Oberthür & Dupont, 2021). Emissions resulting from human and economic activities increase the greenhouse gas concentration in the atmosphere, cause the earth to warm up and cause climate change. According to the report of The Intergovernmental Panel on Climate Change (IPCC), global warming must remain at least 1.5°C to control the negative effects of climate change. However, with the continuation of the current policies, it is estimated that the earth’s temperature will be 3°C until 2050 (IPCC, 2018). For this reason, the European Union has adopted various policies to reduce greenhouse gas emissions and support energy from clean sources, while being at the forefront of global action against climate change and active in international climate negotiations. However, the EU has not been able to convincingly reduce its greenhouse gas emissions. As greenhouse gas emissions in transport increase, coal use in electricity systems continues to play a permanent role. In this case, it is known that the first steps were taken in December 2019, when Ursula Von Der Leyen, the European Commission presidential candidate, suggested that important steps should be taken to expand and strengthen the EU climate policy. The EU has proposed the European Green Deal (EGD) until 2050 and the European Climate Law, which will require climate neutrality. In order to achieve this goal, a comprehensive policy framework covering climate, energy, environment, industry, economic and social aspects, which is the main subject of the EGD, was deemed necessary. 1. Introduction At this point, Von der Leyen’s suggestions with the Constitutional Court include increasing the EU’s 2030 emission reduction target from 40 percent to 55 percent, introducing a carbon limit tax, preparing a Sustainable European Investment Plan, and the European Investment Bank’s proposals. (EIB) into a climate bank, expansion of the EU emissions trading system (ETS) and development of a new industrial policy for Europe (Von der Leyen, 2019). Carbon pricing, sustainable investment, industrial policy, and a fair and competitive transformation draw attention as the most important factors here. In order to ensure a fair competition environment especially for European and foreign producers and to have a significant impact on the environmental policies of third countries, the EU wishes to apply a carbon price to all energy-intensive imports (Bochkarev, 2020). In particular, carbon dioxide, which has a rate of 70% in greenhouse gas emissions, is considered the most important gas that needs to be reduced. Although large economies, especially energy production and industrial sectors, show a development dependent on Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy 3 carbon dioxide emissions, it is necessary to limit carbon emissions, which have such a negative effect on climate change (Ustun, 2021). It therefore aims to establish a meaningful carbon price for all sectors by strengthening the EU emissions trading system (ETS) and forcing EU countries to raise the price of emissions not covered by the ETS. It is essential to put a price on all emissions because pricing encourages all interested parties to reduce their greenhouse gas footprints. Without such pricing, there is no effective reduction of emissions through subsidies or other climate policy measures such as standards. Therefore, it becomes necessary to apply a reasonable price for all greenhouse gas emissions (Claeys et al., 2022). However, it is stated that such an application may produce different results. In particular, “green taxation” can create social and political problems in countries that are fragile and rely heavily on hydrocarbon revenues to finance their budgets. Such a situation may cause third countries to respond with additional tariffs or various trade barriers to equalize competition. In addition, if the US applies a carbon tax to the shale gas sector, there may be trade problems between the USA and the EU (Bochkarev, 2020). For this reason, the carbon tax applied in accordance with the WTO rules should be a protective measure for all countries (GATT, 1947). 1. Introduction On the other hand, it is considered necessary to prepare a carbon border tax to provide a robust mechanism against carbon leakage. It is therefore necessary to introduce other tools first, including subsidies for low-carbon exports and stricter environmental standards that importers must comply with to access the EU market. On the other hand, in this context, the necessity of adopting a sustainable investment strategy and supporting green investment by shifting funds to this purpose to balance the increasing costs encountered due to high carbon prices has also manifested itself (Claeys et al., 2022). However, the EGD is not only a matter of concern to the EU, but also binds all countries that have political, social, economic, or financial relations with the EU. In particular, the EGD aims to lead the way in the transformation of greenhouse gas emissions in countries other than the EU. At this point, the necessity of making commercial agreements to reduce greenhouse gas emissions in Turkey, which has an intense trade volume with the EU, and to ensure that the carbon limit is kept in balance by aiming to minimize the risk of carbon leakage has manifested (Sahin et al 2021) Turkey represents 3.6% of the EU’s total goods trade with the world in 2020. With this ratio, it is the 6th largest trading partner of the EU. While the EU is Turkey’s largest export and import partner by far, it is also the main source of investment (European Commission, 2022). 4 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models Table 1. 2018-2021 Trade in Goods, € billions (European Commission, 2022) Table 1. 2018-2021 Trade in Goods, € billions (European Commission, 2022) Year EU imports EU exports Balance 2018 66.9 69.2 2.3 2019 69.8 68.3 -1.6 2020 62.4 70.1 7.7 2021 78.0 79.3 1.3 As can be seen in Table 1, the total trade in goods between the EU and Turkey reached € 157.3 billion in 2021. Therefore, as stated in the EGD, between the EU and Turkey, which has such a high trade volume, it was necessary for the EU to reach an agreement with Turkey to prevent carbon emissions from leaking into Europe. In particular, Turkey needs to make its production structure suitable for green transformation by purifying it from pollution and carbon emissions. 1. Introduction At this point, it is observed that the share of electricity generation in Turkey’s total greenhouse gas, which is calculated as 506 million tons of CO2 equivalent in 2019, is 140 million tons (28%) (Ecer et al., 2021). On the other hand, it is concluded that it would be more correct to start green transformation in the field of energy, considering that the largest share in total greenhouse gas emissions in Turkey in 2020, with 70.2% as CO2 equivalent, is energy-based emissions (TUIK, 2022). Therefore, the changes that the European Green Deal and the European Union will implement in many sectors directly concern the countries that have trade relations with the EU. Turkey, one of these countries, needs to make the necessary arrangements in order not to disrupt its trade relations with the EU countries, which are among the most important trade partners. For all these reasons, it is estimated that estimating greenhouse gas emissions based on the decision tree model for Turkey within the framework of the European Green Deal will eliminate the gap in the literature. In the first part of the study, information about the European Green Deal was shared and the operation of Carbon Border Adjustment Mechanism (CBAM) was explained. Then, the possible effects of the CBAM on Turkey were given. A literature review was conducted on studies examining the relationship between environmental taxes and pollution, which play an important role in reducing environmental pollution and the effects of the European Green Deal on Turkey were explained. In the last part, the findings obtained after the model and methodology were evaluated and various policy recommendations were made. Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 5 Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 5 2. European Green Deal 2. European Green Deal The global warming that the world is going through and therefore climate change has damaging consequences in many areas, especially in food. In particular, the increasing trend of greenhouse gas emissions after the industrial revolution is dragging the world towards the point of no return. The map in Figure 1 shows the per capita CO2 emissions of the world’s countries. Overall, countries such as Canada, the United States, Russia produce more than 10 tons of emissions per person. Figure 1: CO2 emissions per capita (tonnes) (https://ourworldindata.org/co2-emissions, 2022) Figure 1: CO2 emissions per capita (tonnes) (https://ourworldindata.org/co2-emissions, 2022) EU countries and Turkey are again high but have CO2 emissions of less than 10 tons per person. Environmental problems that arise because of high emission values have always been put on the second plan due to the growth concerns of developed and developing countries. However, with the negative effects of global temperature increase on human health, biodiversity, nature, climate, etc. are clearly felt, permanent plans and programs have started to be developed instead of palliative solutions. The process that started with the Kyoto protocol continued with the Paris climate agreement, to which Turkey is a party, and then expanded with the EGD, one of the most comprehensive climate programs. The EGD can be expressed as a solution to the problem of Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 6 global climate change. With a modern economic transformation that ensures competitive and efficient sourcing, it aims to reduce CO2 emissions by 55% by 2030 and to achieve climate neutrality by 2050 (EC, 2019). European economy, social life, urban life, agricultural, energy and environmental policies contain a comprehensive change strategy. These strategies are expressed in the European Union Commission report as follows (EU, 2019): • Within the framework of the EU neutral climate objective, despite the 61% growth rate of the economy between 1990 and 2018, it managed to reduce greenhouse gas emissions by 23%. By 2050, this rate is targeted to be 60% or more. In this context, developing carbon pricing is an important and effective strategy for reducing greenhouse gas emissions. • Construction, use and renovation of buildings require significant amounts of energy and mineral resources. In addition, the energy consumption of buildings is over 40%. 2. European Green Deal In order to reduce energy consumption and more efficient processes in construction activities, EU member states have initiated a “wave of renewals” for private and public buildings. In this context, the use of different energy sources is supported to ensure energy efficiency. • One in 3 of the EU’s greenhouse gas emissions comes from transport and the rate is steadily increasing. For a neutral climate, it is aimed to reduce greenhouse gas emissions by 90% by 2050. In this direction, it is planned to develop multimodal transportation networks. • Agricultural activities cause water, air and soil pollution and lead to climate change and overuse, especially by reducing biodiversity. Within the scope of the farm-to-fork strategy, new opportunities have been created in the food chain. It is aimed to create awareness in the field of food chain and sustainable food, which includes technology and scientific discoveries. • New technologies, sustainable solutions and disruptive innovation are critical to achieving the objectives of the European Green Deal. To maintain its competitive advantage in clean technologies, the EU needs to significantly increase the large-scale distribution and representation of new technologies across sectors and across the single market, creating new innovative value chains. Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 7 • National budgets play a key role in the transition. Greater use of green budgeting tools will help shift public investment, consumption, and taxation toward green priorities and away from harmful subsidies. • National budgets play a key role in the transition. Greater use of green budgeting tools will help shift public investment, consumption, and taxation toward green priorities and away from harmful subsidies. The EU has created an emissions trading system to reduce greenhouse gas emissions. ETS is one of the cornerstones of the EU greenhouse gas emissions reduction strategy. (EC, 2021a). This system works within the scope of the “upper limit and trade” system. The upper limit is set on the total amount of certain greenhouse gases that can be emitted by the facilities covered by the system. As the total emission rate decreases, so does the upper limit. In practice, carbon is offered to the sectors free of charge, but when the companies operating in these sectors exceed the specified upper limit rate, they buy the excess amount from the market (TUSIAD, 2020). 2. European Green Deal This system, which is currently only applied with EU countries, will be expanded within the scope of international trade, and will also include countries that trade with EU countries. This system, called Carbon Border Adjustment Mechanism (CRAM), aims to reduce carbon leakage from the EU and plans to implement new taxes and non-tariff barriers in international trade in accordance with World Organization rules (TUSIAD, 2020; UNCTAD, 2021). In this context, the average carbon emissions of the companies in the EU will be determined according to the sectors and taxation will be made for the part above this average. However, the fact that this practice does not cover non-EU countries causes a problem such as “carbon leakage” (EC, 2021 b). When the studies in this field are examined, it is seen that carbon leakage does not create a significant problem for the EU industry. Since the application is only valid for EU member states and does not cover the countries that trade with these countries, the chances of success of the application are reduced. For this reason, it has been decided to introduce tax/tariff barriers in non-EU countries due to excessive carbon emissions with the CRAM. The functioning of the CRAM is shown in Table 2: 8 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 8 Table 2: How the CRAM Mechanism Works (Lowe, 2021) CRAM Models Definition Excise Tax Imposition of a Carbon tax on products that emit carbon during production, regardless of where the product comes from. For example, a tax of 50 euros per ton of CO2 emitted, regardless of where the product is produced. As with the application of carbon VAT, the tax will be paid at the time of importation. Production Fee Imposing a carbon tax on European producers. In this case, a tax occurs compared to the carbon cost covered by an average EU- based company producing a similar product. For example, the carbon tax per ton of iron (50 euros), which is determined considering the average costs in domestic production, will also apply to imported iron. Connecting with ETS If the de facto EU local carbon price is determined by the cost of ETS permits, CRAM importers will pay import duties compared to costs determined by the average production of EU-based firms. 2. European Green Deal For example, if an average EU iron producer has 2 units of ETS permits, the same unit quantity of imported iron will also face the same tax rate. Table 2: How the CRAM Mechanism Works (Lowe, 2021) Considering the definitions in Table 2, ETS will be moved to the international level and companies located in third countries will be subject to carbon tax as much as the tax paid by companies operating in EU countries. Therefore, countries that trade with the EU need to regulate their carbon emissions. Third countries will either introduce practices to reduce emissions or introduce taxes within the framework of the EU CRAM system. The implementation of the CRAM will cover risky sectors in the first phase. These sectors are cement, iron and steel, aluminum, electricity, and fertilizer. Importers of these goods covered by the CRAM will apply to the institutions in their country and obtain a SG certificate. If importers declare that a carbon tax has been paid during the production of products from third countries, the CRAM tax will not be levied on these firms (EC, 2021 b). The top 20 countries engaged in foreign trade in the specified sectors with EU countries are listed in Figure 2. Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 9 Figure 2: Top 20 Countries Where the EU Trade the Most by Risk Sectors (UNCTAD, 2021) Figure 2: Top 20 Countries Where the EU Trade the Most by Risk Sectors (UNCTAD, 2021) According to the figure, the top 3 countries that will be most affected by the CRAM system are Russia, China, and Turkey. The trade share of these countries in risky sectors is quite high. Therefore, it is important that the sectors in these countries determine the upper limit of their emission rates according to the EU level. At this stage, the impact of the CRAM depends on the trade model, the carbon intensity of the products produced by countries and the carbon policies of EU trading partners (EC, 2021b). One of the main criticisms of carbon regulation at the border is the debate that it will reduce free trade. In this context, it is stated that carbon regulation at the border is a hidden foreign trade tax. 2. European Green Deal In the country commission report submitted within the scope of the impact analysis of the European Union Carbon Regulation at the Border of the Republic of Turkey, it was stated that unilateral actions such as the carbon limit are regulations related to international trade risk that create trade barriers and are used as protectionists, and may lead to unilateral trade retaliations, especially in times of trade wars. Therefore, it is stated that the trade divergent effects of the CRAM should be eliminated. Another criticism of the implementation of the ministry is that the CRAM should be stretched according to the economic characteristics of the countries. In particular, it is stated that the special conditions of developing countries should be taken into consideration in practice. Another criticism is the effectiveness of the application. However, the effectiveness of the application and its effects 10 Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models on the climate will be clearly demonstrated in the following processes. 3. Dilemma for Turkey Turkey is a party to the Paris agreement and has signed the agreement in 2021, becoming one of the countries that take active measures against global climate change. Within the scope of this agreement, Turkey has declared its greenhouse gas emissions as a reduction from an increase of up to 21% in the “Intended National Contribution” declaration foreseen until 2030. In addition, Turkey aims for net zero emissions by 2053 (GRB, 2021). Therefore, in this respect, it is seen that Turkey has a decisive stance on climate change with the agreements to which it is a party and its domestic legal practices. However, at this stage, some economic, cultural, and legal practices should be implemented. Due to Turkey, EU membership process, Customs Union, geographical proximity, etc., EU countries trade the most. In Table 3, the trade shares of the country groups to which Turkey exports the most are included. 2. European Green Deal Table 3: Share of Turkey’s export countries in all exports (%) (TÜİK, 2021) Country Groups 2021 2020 2019 2018 Table 3: Share of Turkey’s export countries in all exports (%) (TÜİK, 2021) Country Groups 2021 2020 2019 2018 A-European Union (EU 27) 41.32128983 41.27600459 42.42937729 43.70364535 B-Other countries 58.67871017 58.72399541 57.57062271 56.29635465 1-Other Europe (except EU) 23.80945449 24.50000839 23.48109108 23.98346698 2-North Africa 10.09301225 9.259424925 10.40172836 9.940968236 3-Other Africa 5.958142579 6.039680172 5.565763936 5.18738237 4-North America 12.41207518 11.23141135 9.563936607 10.43864885 5-Central America and Caribbean 1.848485504 1.458462764 1.697193732 1.652813272 6-South America 2.718969499 1.935992548 1.760774312 1.877726735 7-Near and Middle East 29.0226674 31.45328122 33.62814241 32.48750138 8-Other Asia 13.03788993 12.8245916 12.87093968 13.33675835 9-Australia and New Zealand 0.853131192 0.840498687 0.751498367 0.827174348 10-Other Countries and Regions 0.246171978 0.456648341 0.278931512 0.267559486 Note: Calculated by the authors using TURKSTAT data. Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 11 When Table 3, which contains data between 2018-2021, is examined, an average of 42% of Turkey’s total exports are made to EU member countries. The share of other countries is on average about 58%. Since these data are taken into consideration, with the launch of the CRAM, Turkey ‘s exports will be significantly affected by the carbon tax. At this stage, it is necessary to add the advantages of Turkey ‘s membership in the Customs Union. The Customs Union allows the free movement of goods and services in Turkey and the EU countries. The place of the carbon tax to be introduced with the implementation of the CRAM within the scope of the Customs Union is controversial. Turkey draws attention to its rights arising from this agreement and states that the trade divergent effect of the practice should be eliminated. In addition, as mentioned before, the implementation of the CRAM according to the specific conditions of the countries is also recommended by Turkey. Looking at the emission rate in general and the countries with which the EU trades, it is clear that the CRAM will impose an additional cost on Turkey, but this cost can be reduced by the relevant regulations within the country. 4. Literature The available literature on the European Green Deal and its implications is very limited. Even if the studies on Turkey have been intensified recently, they are mostly in the form of literature reviews. This is why the literature will be examined under two basic parts. In the first part, studies investigating the effects of environmental taxes on pollution, and the second part on the effects of EGD on Turkey are included. The Relationship Between Environmental Taxes and Pollution Kumbaroglu (2003), in his study discussing the effects of environmental taxes on the Turkish economy, states that taxes offer important opportunities in environmental and economic terms. Within the scope of the study, five different tax scenarios were created. The scenarios are built on assumptions of a choice to impose an emissions tax to improve environmental quality, a reduction in energy imports to accelerate economic development, and a replacement for hard coal and lignite with oil and gas to reduce pollution. The results obtained point to the importance of energy saving and indicate that the effect on national income varies according to the type of greenhouse gas emissions that are taxed. For example, in the study, it is stated that energy exports decrease with the addition of tax on NOx and that national income will increase with this effect, while a tax on SO2 reduces national income. Acar and Yeldan, (2016), in their study, state that by reducing the Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 12 incentive of Turkey’s use of coal for energy, national income will decrease due to dependence on energy imports, but at the same time it will reduce CO2 emissions and improve environmental quality. Bouzaher et al. (2015) designed two different scenarios in their work. The first scenario considers environmental taxes on the energy use of households and industry, while the second scenario examines the process of introducing an additional sectoral tax on CO2 emissions and transferring these tax revenues to green innovation. Accordingly, in the first scenario, national income decreases, but in the second scenario, national income increases. Yeldan and Voyvoda (2015) state in their studies that the implementation of carbon taxes in Turkey and the redirection of the renewable energy investment fund to be created with these taxes to renewable energy sources increase energy efficiency. Ding et. al. (2019) stated that environmental taxes reduced pollution by about 28% in countries with high pollution rates. Likewise, Polat and Ergün (2021) have concluded that environmental taxes reduce pollution emissions in their studies for 21 EU countries. Hao et. In their study investigating the effects of green growth (2021), they found that environmental taxes reduced CO2 consumption. Born, meat. In their study, al. (2022) examined the effects of green growth and environmental taxes between 1994 and 2018 for 25 environmentally friendly countries. The Relationship Between Environmental Taxes and Pollution At the end of the study, while a two-way causal relationship was found between economic growth and environmental pollution, no relationship was found between pollution and environmental taxes. Similarly, Yavuz and Ergen (2022) examined environmental taxes and environmental pollution for G20 countries in their studies and concluded that environmental taxes do not influence pollution. In contrast, Damirova and Yayla (2020) examined the relationship between environmental pollution and microeconomic variables using data from 1995 to 2016 for Denmark, Hungary, İtaly, Malta, Netherlands, Portugal, Slovakia, Switzerland England and Turkey. The study also looked at the effectiveness of environmental taxes. Accordingly, environmental taxes do not have a statistically significant effect on environmental pollution. The Impact of the European Green Deal on Turkey The Relationship Between Environmental Taxes and Pollution According to the results, environmental taxes are one of the key factors to reduce CO2 emissions. Ozkaya (2022) investigated the effects of economic growth and environmental taxes on pollution in terms of European Union countries. At the end of the study, while a two-way causal relationship was found between economic growth and environmental pollution, no relationship was found between pollution and environmental taxes. Similarly, Yavuz and Ergen (2022) examined environmental taxes and environmental pollution for G20 countries in their studies and concluded that environmental taxes do not influence pollution. In contrast, Damirova and Yayla (2020) examined the relationship between environmental pollution and microeconomic variables using data from 1995 to 2016 for Denmark, Hungary, İtaly, Malta, Netherlands, Portugal, Slovakia, Switzerland England and Turkey. The study also looked at the effectiveness of environmental taxes. Accordingly, environmental taxes do not have a statistically significant effect on environmental pollution. incentive of Turkey’s use of coal for energy, national income will decrease due to dependence on energy imports, but at the same time it will reduce CO2 emissions and improve environmental quality. Bouzaher et al. (2015) designed two different scenarios in their work. The first scenario considers environmental taxes on the energy use of households and industry, while the second scenario examines the process of introducing an additional sectoral tax on CO2 emissions and transferring these tax revenues to green innovation. Accordingly, in the first scenario, national income decreases, but in the second scenario, national income increases. Yeldan and Voyvoda (2015) state in their studies that the implementation of carbon taxes in Turkey and the redirection of the renewable energy investment fund to be created with these taxes to renewable energy sources increase energy efficiency. Ding et. al. (2019) stated that environmental taxes reduced pollution by about 28% in countries with high pollution rates. Likewise, Polat and Ergün (2021) have concluded that environmental taxes reduce pollution emissions in their studies for 21 EU countries. Hao et. In their study investigating the effects of green growth (2021), they found that environmental taxes reduced CO2 consumption. Born, meat. In their study, al. (2022) examined the effects of green growth and environmental taxes between 1994 and 2018 for 25 environmentally friendly countries. According to the results, environmental taxes are one of the key factors to reduce CO2 emissions. Ozkaya (2022) investigated the effects of economic growth and environmental taxes on pollution in terms of European Union countries. The Impact of the European Green Deal on Turkey When the academic studies on the effects of the CRAM on Turkey are examined, it is seen that the environmental impact is positive. In the TUSIAD (2020) report prepared by Yeldan et al., two different scenarios related to the effects of EGD were studied. In the first of these scenarios, the price per ton of carbon is calculated as 30 euros, and in the second as Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy 13 50 euros. According to the data obtained, when 30 euros are taxed, GDP increases by 5.7%, while the tax application of 50 euros increases by 6.6%. As a result of taxation, greenhouse gas emissions decrease by 16.5% and 15%, respectively. Therefore, the implementation of the CRAM both offers an opportunity for environmental improvement for Turkey and triggers the increase in national income. However, unlike these studies, according to a study conducted by UNCTAD (2021), it is stated that the introduction of an additional CO2 tax in the sector will have an impact on the iron and steel sector in Turkey and that the competitiveness in this sector will decrease. In addition, it is stated that both price increase will occur in other sectors and competitiveness will be adversely affected. Turker and Aydin (2022) similarly state in their studies that environmental sustainability will be positively affected if EGD strategies are supported by national policies. When these studies are evaluated, it is seen that the implementation of the CRAM reduces greenhouse gas emissions for Turkey, but the impact on the country’s economy depends on national policies. 5. Model and Methodology The application model of this study is to investigate the effect of tax and other variables on the emission value of Turkey and to predict the emission in the next 5 years to observe the effect of environmental tax. In practice, M5p algorithm, one of the data mining decision tree algorithms, was used. The M5p algorithm was preferred because it provides linear regression models for the past and future and estimation of values in the data set. 5.2. Decision Trees Decision tree methods, one of the data mining methods, are a forecasting model that can be used in classification and regression problems. When a decision tree is used for classification tasks, it is referred to as a classification tree. When used for regression tasks, it is called a regression tree. Decisions trees clearly define the functional dependencies of the target feature on descriptive attributes (H. Dahan et al. 2014; Maimon, O. Z., & Rokach, L. 2014). 5.1 Data The data used in the study were obtained by literature review. The dataset covers the period 1994-2020 and consists of environmental tax, exports, energy consumption, waste sector CO2 emissions, agricultural sector CO2 emissions, and total CO2 emissions. The data are presented in Figure 3. 14 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 14 Figure 3: Data Figure 3: Data When the time series of the variables in Figure 3 are examined, it is seen that the emission rates in Turkey have increased continuously since 1994. In addition, while exports, energy consumption and the number of vehicles increase over time, there are fluctuations in the trend of environmental taxes, and there is a decrease in environmental taxes, especially in the later period. 5.3 M5p Model Trees (M5) was proposed by Quinlan in 1992 as a tree algorithm for predicting continuous variables. They were led by Wang and Witten (1998) to propose the M5p algorithm, which is categorical and has the 15 Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy advantages of being able to deal with continuous and incomplete variables (Frank, Wang Y, Inglis, Holmes and Witten, 1998; Barros, Basgalupp; Lin, L., Wang, Q., & Sadek, A. W, 2016). The M5P algorithm consists of four main steps. In the first step, the input space is divided into several subspaces to form a tree. Subspace variability from root to node is minimized using the division criterion. Construction of the tree is done using the standard deviation reduction (SDR) factor that maximizes the expected error reduction at the node as follows (Behnood, A., Behnood, V., Gharehveran, M. M., & Alyamac, K. E. 2017).: T=number of samples; Ti=number of samples that represent the sample that have the potential rise rise sd=standard deviation. sd=standard deviation. In the second step, a linear regression model is created by using the data associated with that subspace in each subspace. Here, a pruning technique is used to solve the overtraining problem. During the pruning process, smoothing is done to compensate for sharp discontinuities between adjacent linear patterns. The smoothing process combines all the patterns from leaf to root to create the final pattern of the leaf. In this process, the estimated value of the leaf is filtered out on its way back to the root. The combination of this value with the value predicted by linear regression for that node is as follows: E’: is the estimated value to the next top node, E’: is the estimated value to the next top node, e: is the prediction passed from below to the current node, e: is the prediction passed from below to the current node, a : is the value predicted by the model at this node, a : is the value predicted by the model at this node, n is the number of training samples reaching the following node and k is a constant. 6 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models 5.4. Model Evaluation Criteria In order to evaluate the model created in the study, the most common accuracy metrics of regression models: MAE (Mean Absolute Error, MAE), RMSE (Root Mean Square Error, RMSE), MAPE (Mean Absolute Percentage Error, MAPE) were used. Accuracy metrics are shown in Table 4. Table 4: Accuracy metrics MAE = RMSE = 6. Empirical Results 6. Empirical Results In the data set used, the value with the highest SDR value was determined as the number of vehicles and LM1 and LM2 predictive models were created. Table 5: Lineer Models Table 5: Lineer Models Number of Vehicle <= 11686609.5 : LM1 (12/2.617%) Number of Vehicle > 11686609.5 : LM2 (15/3.999%) LM num: 1 emissions = + 0.9457 * Energy + 67.4095 LM num: 2 emissions = + 0.8816 * Energy + 58.3739 When Table 5 is examined, it is seen that the CO2 amounts originating from the energy sector are effective in estimating the CO2 emission rate. With the number of vehicles exceeding 11686609.5, the energy coefficient in the LM2 model decreased to 0.8816. Table 6: Model Evaluation Criteria MAPE 0,0112 RMSE 0,015 MAE 3,8753 Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 1 Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy \| 1 17 In Table 6, the values of the M5p model created in the study according to the performance evaluation criteria are included. When the values in Table 6 are examined, it can be said that the model is successful. In Figure 4, the realized and estimated values of the emission value in the model are included according to the years. When Figure 4 is examined, it can be said that the emission value is constantly increasing, and this value will reach 597,792 metric tons in 2025. Figure 4: CO2 Emissions Actual and Prediction According to Tax Figure 4: CO2 Emissions Actual and Prediction According to Tax In the prospective forecast model, where the tax value created in the application increased by two times and the non-tax values in the data set were fixed, the emission value was estimated as 523,443. It can be said that the tax value does not help to reduce the emission value. Conclusion and Policy Recommendation The European green deal includes various arrangements both within the EU and internationally, with European countries systematizing several measures against climate change. The emissions trading system, which is primarily implemented within the borders of the EU, aims to improve and minimize emission values. In order to improve the effectiveness of this implementation, the emissions trading system will also be made available to other countries that trade with the EU, bringing it closer to the goal of minimizing CO2 emissions. Turkey is also one of the countries that will be included in the emissions trading system because it is among the countries that trade the most with the 18 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models EU. At this stage, it makes the necessary arrangements to be included in this system with the road map (2016) prepared by the Ministry of Environment and Urbanization of Turkey. Turkey is expected to start implementing its emissions trading system by the end of 2022. However, within the framework of the EU green deal, carbon application at the border points to an important problem for Turkey. Turkey draws attention to its rights arising from the customs union regarding emission regulation (Ministry of Trade, 2020). The implementation of the application is likely to increase the costs of the companies and therefore there will be a decrease in foreign trade with the EU due to these costs. In the study, the effects of environmental taxes on emission trading in Turkey and the future values of emission values were estimated because of increasing the effectiveness of tax application. For these predictions MAE (Mean Absolute Error, MAE), RMSE (Root Mean Square Error, RMSE), MAPE (Mean Absolute Percentage Error, MAPE) methods were used. According to the data obtained, environmental taxes currently have no effect on emission values in Turkey. This result is thought to be since environmental taxes are not effective. Especially in developing countries, taxes such as environmental taxes are seen as costs that reduce production and are not applied effectively. In addition, according to the analysis made under the assumption of increasing taxes within the scope of the emission trading system, it was concluded that the effects of taxes were limited in the next 5 years. Conclusion and Policy Recommendation However, it is estimated that this system, which is not effective in the short term, may reduce the emission values of the companies with R & D and innovation studies in the long term and thus the emission values will decrease. Therefore, in order to effectively implement climate change policies, it is important for Turkey to develop more effective and preventive policies, to conduct audits on a firm basis, and to direct companies to R & D and innovation studies in order to reduce emission values. Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy 19 References References Acar, S., Kitson, L. & Bridle, R., (2015). Subsidies to Coal and Renewable Energy in Turkey. Global Subsidies Initiative Report, March 2015, IISD. 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Centre for European Reform, 21 21 Orhan Ecemis / Asli Ozpolat / Ferda Nakipoglu Ozsoy Retrieved from https://www.cer.eu/sites/default/files/pbrief_cbam_ sl_21.4.21.pdf (Accessed: 28.02.2021). Oberthür, S., & Dupont, C. (2021). The European Union’s International Cli- mate Leadership: Towards a Grand Climate Strategy? Journal of European Public Policy, 28(7), 1095–1114. https://doi.org/10.1080/13501763.20 21.1918218 Our World in data (2022). CO2 and Green Gas Emissions. Retrieved from ht- tps://ourworldindata.org/co2-and-other-greenhouse-gas-emissions (Ac- cessed: 01.03.2022). Ozkaya, M.H. (2022). Ekonomik Büyüme ve Çevre Vergi Gelirlerinin Karbon- dioksit Emisyonu Üzerindeki Etkisi: AB Ülkeleri Örneği. International Journal of Applied Economic and Finance Studies, 7(1), 128-139. Polat, M. A. & Ergün, S. (2021). Kirlilik Emisyonlarının Azaltılmasın- da Çevre Vergilerinin Rolü: AB Ülkeleri Örneği, İzmir İktisat Dergisi, 36(2):379-397. Rebuplic of Turkey Ministry of Environment, Urbanization and Climate Chan- ge (2020). Turkey’de Sera Gazı Emisyon Sisteminin Kurulmasına Yönelik Yol Haritası. Retrieved from https://pmrturkiye.csb.gov.tr/wp-content/ uploads/2020/12/Turkiyede-Sera-Gazi-Emisyon-Ticareti-Sisteminin-Ku- rulmasina-Yonelik-Yol-Haritasi.pdf (Accessed: 11.12.2022). Rebublic of Turkey Ministry of Trade (2020). 1 Gaziantep Üniversitesi, İktisadi ve İdari Bilimler Fakültesi, İktisat Bölümü, İktisat Politikası Anabilim Dalı Araştırma Görevlisi Özet Gelir dağılımı, iktisat bilimi başta olmak üzere birçok bilim ve disiplinin odağında yer alan önemli ve güncel bir çalışma konusunu oluşturmaktadır. Tarihsel süreç içerisinde birçok dinamiğin etkilediği gelir dağılımı, günümüzde teknolojik gelişmelere bağlı olarak değişim göstermektedir. Ülkelerin siyasi, ekonomik ve toplumsal yapıları da gelir dağılımında etkili olan majör etmenlerden birkaçıdır. Gelirin bölüşümünde adaletin sağlanması gerek ekonomik gelişmenin gerekse toplumsal huzurun tesis edilmesinde pay sahibidir. Buradan hareketle bu çalışma Gelir dağılımı eşitsizliğinin ekonomik, sosyal ve siyasi boyutlarının bütünü göz önünde bulundurularak, Türkiye’de gelir dağılımı sürecine ilişkin bir görünüm oluşturmaktadır. Çalışmada öncelikle gelir dağılımına ilişkin kavramsal çerçeve irdelenmiştir. Daha sonra Türkiye’de yakın dönemde meydana gelen sosyo-ekonomik ve siyasi gelişmelere yer verilerek gelir dağılımına olası etkileri göz önünde bulundurulmuştur. Son olarak gelir dağılımının ölçümünde kullanılan yaygın göstergelerden Gini katsayısı Türkiye verilerine yer verilerek, Türkiye’de gelir dağılımına ilişkin genel görünüm sunulmuştur. References Views of the Government of Turkey on the Carbon Border Adjustment Mechanism Within the Fra- mework of the Inception Impact Assessment Retrieved from https:// ticaret.gov.tr/data/5f901d8813b8760d9ce54aeb/AB%20S%C4%B1n%- C4%B1rda%20Karbon%20D%C3%BCzenleme%20Mekanizmas%- C4%B1%20Etki%20Analizi-%C3%9Clke%20G%C3%B6r%C3%B- C%C5%9F%C3%BC_Ek_Turkey%20Views%20on%20CBA.pdf.pdf (Accessed: 11.12.2022). Rebublic of Turkey Ministry of Trade, (2021). Sınırda Karbon Düzenleme Me- kanizması Etki Analizi Çerçevesinde Komisyon’a iletilen Ülke Görüşü. Retrieved from AB Sınırda Karbon Düzenleme Mekanizması Etki Ana- lizi-Ülke Görüşü_Ek_Turkey Views on CBA.pdf.pdf (ticaret.gov.tr) (Ac- cessed: 13.03.2022). Rebublic of Turkey Ministry of Trade, (2021) Yeşil Mutabakat Eylem Planı, retrieved from https://ticaret.gov.tr/data/60f1200013b876eb28421b23/MUTABAKAT%20 YE%C5%9E%C4%B0L.pdf (Accessed: 08.03.2022). Şahin, G., Taksim, M. A., ve Yitgin, B., (2021). Effects of the European Green Deal on Turkey’s Electricity Market. İşletme Ekonomi ve Yönetim Araştır- maları Dergisi, 4(1), 40-58. 22 22 \| Evaluation European Green Deal for Türkiye: Based on The Decision Tree Models The General Agreement on Tariffs and Trade, 1947, retrieved from https:// www.wto.org/english/docs_e/legal_e/ gatt47_01_e.htm. (Accessed: 21.11.2022). Türkiye Istatistik Kurumu, (2022), retrieved from https://data.tuik.gov.tr/Bulten/Index?p=Greenhouse-Gas-Emissions-Statisti- cs-1990-2020-45862 (Accessed: 21.11.2022). Türker Ozhan, Y. & Aydın, A. (2022). How Ready is the Turkish Legislation for the Green Deal? Energy and Climate Change, 3 (2022) 100084. TÜSİAD. (2020). Ekonomik Göstergeler Merceğinden Yeni İklim Rejimi. Eri- şim adresi Ekonomik Göstergeler Merceğinden Yeni İklim Rejimi Raporu (tusiad.org) (Accessed: 13.03.2022). UNCTAD, (2021). European Union Carbon Border Adjustment Mecha- nism: Implications for Developing Countries. Retrieved from https:// unctad.org/webflyer/european-union-carbon-border-adjustment-mecha- nism-implications-developing-countries (Accessed: 13.03.2021). USTUN, K. T. (2021). Yeni Bir Dönemin Başlangıcı: Avrupa Yeşil Mutabakatı Ve Türk Çevre Hukuku Ve Politikalarına Etkileri. Memleket Siyaset Yöne- tim, 16(36), 329-366. Von der Leyen, U., (2019). A Union that Strives for More: My Agenda for Europe, Political Guidelines for the Next European Commission 2019- 2024, retrieved from https://ec.europa.eu/commission/sites/ beta-po- litical/fi les/political-guidelines-next-commission_en.pdf (Accessed: 08.03.2022). Yavuz, E. & Ergen, E. (2022). Çevre Vergilerinin Çevre Kirliliği Üzerindeki Et- kisi: Seçilmiş G20 Ülkeleri Üzerine Bir Uygulama, International Journal of Public Finance, 7(1), 113-136. Yeldan, E. & Voyvoda E. (2015). Türkiye için Düşük Karbonlu Kalkınma Yol- ları ve Öncelikleri. WWF- retrieved from http://awsassets.wwftr.panda.org/downloads/20151007_turkiye_icin_ duuk_karbonlu_kalknma_yollar_ve_oncelikleri_rapor.pdf (Accessed: 13.03.2022). Bölüm 2 Türkiye’de Gelir Dağılımına İlişkin Görünüm Bengü Kayapalı Aslan1 Bengü Kayapalı Aslan1 Giriş Türkiye’de gelir dağılımına ilişkin ilk çalışmalar 1960’lı yılların başında yapılmıştır. Bu noktada 1959 yılında Nejat Bengül tarafından hazırlanan “Gelir Dağılımı Araştırması İçin Öneriler” başlıklı çalışma, milli gelirin bölüşümüne yönelik olarak yapılan ilk çalışma olarak kabul edilmektedir (Yücel, 2011: 117). Bu çalışma Stanford üniversitesinde yapıldığı için Devlet Planlama Teşkilatı himayesinde Çavuşoğlu ve Hamurdan tarafından 1963 yılında yapılan çalışma da gelir dağılımına yönelik ilk çalışma olarak nitelendirilmektedir (Acun, 2018: 82). Toplamda 327.000 gelir vergisi mükellefinin verisinden yararlanılarak yapılan çalışmada en alt %20’lik grup https://doi.org/10.58830/ozgur.pub401.c1750 https://doi.org/10.58830/ozgur.pub401.c1750 23 23 Türkiye’de Gelir Dağılımına İlişkin Görünüm 24 gelirin %4,5’ine sahipken, en üstteki %20’lik grubun gelirin %57’sine sahip olduğu görülmüştür (Çavuşoğlu ve Hamurdan, 1966). Gelir dağılımına ilişkin yapılan ilk çalışmadan 2023 yılı TUİK verilerine kadar gelinen süreçte amaçlanan gelir dağılımındaki eşitsizliğin ölçülerek, gelir dağılımındaki adaletin sağlanması noktasında politikalar üretmek ve çözüm önerileri sunmaktır. Gelir dağılımındaki eşitsizlik, ekonomi bilimi başta olmak üzere birçok disiplinin ve bilim dalının önemli bir araştırma konusunu oluşturmaktadır. Özellikle Dünya’da yoksulluğun azaltılması ve sürdürülebilir kalkınmanın azaltılması gibi önemli hedefler içeren Birleşmiş Milletler Sürdürülebilir Kalkınma Amaçları (SDG)’nın hedef ve eylem planları arasında da çokça yer alan gelir adaletsizliği hem yerel hem ulusal hem de uluslararası boyutta önemli bir konuyu oluşturmaktadır (UNDP, 2015). Nitekim literatürde kalkınma ile gelir dağılımı arasındaki ilişkiyi inceleyen birçok çalışma yer almaktadır. Bu çalışmalar gelir dağılımı ile ekonomik büyüme arasındaki ilişkiyi küreselleşme, enflasyon, ihracat, beşerî sermaye vb. değişkenleri kullanarak incelemekte ve buna yönelik olarak değerlendirmelerde bulunmaktadır. Bunun yanında gelir dağılımındaki eşitsizliğin, Dünya’da yaşanan savaş, gerilim, göç, toplumsal huzursuzluk vb. birçok olumsuz gelişmenin sebepleri arasında yer aldığı savunusu da yer almaktadır (İlhan, 2015). Gelir dağılımı eşitsizliğinin ekonomik, sosyal ve siyasi boyutlarının bütünü göz önünde bulundurularak, bu çalışma Türkiye’de gelir dağılımı sürecine ilişkin bir görünüm oluşturmaktadır. Çalışmada öncelikle gelir dağılımı kavramı, gelir dağılımının belirleyicileri, gelir dağılımı ölçüm yöntemleri, gelir dağılımı türleri ve gelir dağılımına iktisat okullarının bakışı incelenerek gelir dağılımının kuramsal çerçevesi çizilecektir. Daha sonra Türkiye’de yakın dönemdeki sosyo-ekonomik ve siyasi gelişmeler incelenerek, istatistiksel verilerden yararlanılarak Türkiye’de gelir dağılımına ilişkin görünüm sunulacaktır. 1. Gelir, Gelir Dağılımı Kavramı ve Gelir Dağılımının Belirleyicileri İktisat, maliye ve hukuk literatüründe farklı perspektifler dahilinde tanımlanan gelir kavramı, iktisat literatüründe en genel anlamıyla üretim aşamasında tüketime kadar geçen süre içerisinde meydana gelen ve para ile ölçülebilen kıymetler olarak ifade etmektedir (Dinçer ve Kalelioğlu, 2013: 87). Diğer bir ifadeyle gelir, kişinin varlığı sabit kalması koşuluyla bir dönem içerisinde tüketebileceği mal ve alabileceği hizmetlerin bütünü 25 Bengü Kayapalı Aslan \| Bengü Kayapalı Aslan olarak da tanımlanmaktadır (Aksoy, 1989: 154). Ücret, rant, faiz ve kardan meydana geldiği savunulan gelirin (Karataş, 2019: 55), gelir dağılımından önce tanımlanmasındaki amaç, gelir dağılımı kavramının daha iyi anlaşılması ve gelir dağılımının/gelirin bölüşümünün odağında yer alan olgunun özümsenmesi adınadır. Gelir dağılımı, bir ülkede belirli bir dönemde üretim sonucunda ortaya çıkan milli gelirin bölüşümünü ifade etmektedir. Bu bölüşüm toplumdaki bireyler, gruplar, üretim faktörleri ve bölgeler arasında gerçekleşmektedir (Asandaş ve Işik, 2021: 105). Bir başka ifadeyle belirli bir dönemde elde edilen milli gelirin, ülkedeki paydaşlar arasındaki bölüşümü gelir dağılımın genel çerçevesini oluşturmaktadır. Gelir eşitsizliğindeki artış, dünya genelindeki ekonomi politikası tartışmalarının merkezinde yer almaktadır (Clements, vd., 2016: 21). Bu yönüyle gelir dağılımı hem iktisat hem finans hem de maliye başta olmak üzere birçok bilim ve disiplinin odağına aldığı önemli bir araştırma konusu olarak literatürdeki yerini korumaktadır. Birçok bilimin ve disiplinin üzerinde çalışmalar yürütmekte olduğu gelir dağılımının birçok belirleyicisi vardır. Bunlar; “emeğin dağılımı, cinsiyet yapısı, yaş yapısı, eğitim düzeyi, servetin dağılımı, faktör fiyatları, enflasyon, kayıtdışı ekonomi, dolaylı-dolaysız vergiler ve negatif vergiler” başta olmak üzere sıralanmış olsa da Dünya’da yaşanan gelişmelere bağlı olarak yeni birçok etmen yer almaktadır (Şerbetçi, 2014: 97-104). Kaynak: Yazar tarafından oluşturulmuştur. 2.Gelir Dağılımı Türleri Gelir dağılımı kavramı, bir ülkede belirli bir dönemde üretilen milli gelirin paylaşımını konu almaktadır. Gelirin bölüşümünde pay sahibi olan hanehalkı, sektör, bölgeler ve üretim faktörleri gelir dağılımının tanımlanmasında ve sınıflandırılmasındaki temel kriteri oluşturmaktadırlar. Buna bağlı olarak gelir dağılımı; fonksiyonel gelir dağılımı, sektörel gelir dağılımı, bölgesel gelir dağılımı ve hanehalkı (bireysel gelir dağılımı) olarak dört temel başlıkta incelenmektedir (Çalışkan, 2010: 92). Buradan hareketle gelir dağılımı türlerine ilişkin genel görünüm aşağıda Şekil.1.’de sunulmuştur. 26 \| Türkiye’de Gelir Dağılımına İlişkin Görünüm Şekil.1. Gelir Dağılımı Türleri Şekil.1. Gelir Dağılımı Türleri Kaynak: Yazar tarafından oluşturulmuştur. Fonksiyonel Gelir Dağılımı ksiyonel Gelir Dağılımı öncelikle üretim faktörlerinin üretil Şekil.1. Gelir Dağılımı Türleri Kaynak: Yazar tarafından oluşturulmuştur. 2.1.Fonksiyonel Gelir Dağılımı Fonksiyonel Gelir Dağılımı öncelikle üretim faktörlerinin üretilen/ortaya çıkan gelirden elde ettikleri payı ifade etmektedir. Daha geniş bir ifadeyle bir ülkedeki milli gelirin, milli gelirin oluşumu sürecinde üretime katılarak Bengü Kayapalı Aslan \| 27 27 Bengü Kayapalı Aslan \| Bengü Kayapalı Aslan aldıkları payı içermektedir. Bu gelir dağılımı, üretim sürecindeki ücret, faiz ve kârın dağılımından meydana gelmektedir. Bu sebeple gelirin emek faktörü, sermaye faktörünün fiyatı ve doğal kaynaklar bu sürecin içerisinde bölüşülen değerler arasında yer almaktadır (Pehlivan, 2009-24-25). 2.3.Bölgesel Gelir Dağılımı Bir ülkedeki bölgeler birbirleri arasında ekonomik faaliyetler, coğrafi özellikler, jeopolitik konum, devlet politikaları, vb. birçok faktöre bağlı olarak değişiklikler göstermektedir. Bölgelerin gelişmişlik düzeylerinin ortaya çıkmasında belirleyici olan bu faktörler gelirin dağılımında da etkili olmaktadır. Ülkedeki farklı bölgeler bu değişikliklere bağlı olarak milli gelirden farklı ölçülerde pay almaktadır. Buna bölgesel gelir dağılımı denilmektedir (Çiçen ve Karakuzu, 2021: 157). Diğer bir ifadeyle bölgesel gelir dağılımı, milli gelirin ülkedeki bölgeler arasındaki dağılımıdır. Bu dağılımın, bölgelerdeki gelişmişlik farklılıklarına göre değişiklik göstermesine bağlı olarak da bölgesel eşitsizlik kavramı ortaya çıkmaktadır (Sefil Tansever ve Kent, 2018). Dolayısıyla bölgesel eşitsizliğin çözümünde gelir dağılımının bölgesel bağlamda adaletinin sağlanması büyük ölçüde öneme sahiptir. 2.2.Sektörel Gelir Dağılımı Bir ekonomide yer alan sektörler tarım, sanayi ve hizmetler olmak üzere üçe ayrılmaktadır. Üretilen gelirin tarım, sanayi ve hizmet sektörleri arasındaki dağılımına sektörel gelir dağılımı olarak tanımlanmaktadır. Sektörel gelir dağılımı, temelde üç ana sektör üzerine kurulmuş olsa da gelirin bölüşümü sosyoekonomik gelişmelere bağlı olarak değişiklik göstermektedir. Tarihsel süreç göstermektedir ki gelirin dağılımında daha önceleri tarım sektörü dominant konumdayken sanayi devrimiyle sanayiye daha sonra da hizmet sektörüne doğru kaydığı görülmektedir (Işık, 2006: 123). Bunun yanında ülkelerin gelişmişlik düzeyleri de sektörel gelir dağılımında etkili diğer bir faktörü oluşturmaktadır. Gelişmekte olan ülkelere bakıldığında gelirin tarım sektöründe, gelişmiş ülkelerde ise hizmet sektörüne bağlı olarak dağıldığı bilinmektedir. 2.4.Bireysel Gelir Dağılımı Bireysel gelir dağılımı, hanehalkının bir ülkedeki milli gelirden aldığı payı tespit etmektedir. Hanehalkı olarak ifade edilen tüketici birimleri, fertler, aileler ve diğer grupların bölüştüğü milli gelirin yüzdesel olarak ifadesi bireysel gelir dağılımının göstergesidir. Bir ülkede bireyin milli gelirden aldığı pay, yani bireysel gelir dağılımı ülkede gelir dağılımının adil olup olmadığı noktasında önemli bir göstergedir. Bireysel gelir dağılımı Türkiye’de Gelir Dağılımına İlişkin Görünüm 28 hesaplanırken toplumdaki bireyler, gruplar ve diğer tüketici birimleri ayrım gözetmeksizin hesaplamaya dahil edilmektedir (Sahota, 1978: 2-7). Bu noktada bireysel gelir dağılımının hesaplanmasında eşitlikçi bir yöntem takip edildiği görülmektedir. Bu sebeple, ülkedeki gelir dağılımı eşitsizliğinin çözümündeki arayışlar da bireysel gelir dağılımı başvurulan bir kaynak olmalıdır (Işık, 2006: 122). 3.Gelir Dağılımı Ölçümünde Kullanılan Metotlar Gelir dağılımının ölçümü, gelir dağılımına bağlı olarak ortaya çıkan toplumsal eşitsizliklerin çözümünde yararlanılması gereken önemli bir veriyi oluşturmaktadır. Bu bağlamda gelir dağılımının ölçümünde birden çok yöntem kullanılmaktadır. Lorenz eğrisi, Gini katsayısı, yüzdelik dilimleme yöntemi, değişim aralığı, Yüzde Paylar Analizi, Kuznets Katsayısı, Atkinson İndeksi ve değişim katsayısı gelir eşitsizliğini ölçmek amacıyla kullanılan başlıca ölçüm teknikleri olarak literatürde yer almaktadır (Çalışkan, 2010: 94). 4.1980 Sonrasında Türkiye’deki Sosyo-Ekonomik Gelişmeler 1980 yılı itibariyle yaşanan gelişmeler, Dünya’da ve Türkiye’de önemli yapısal dönüşümler meydana getirmiştir. Petrol krizi sonrasında Dünya’da enerji kaynaklarının paylaşımı üzerine teoriler ve stratejiler geliştirilirken, ortaya çıkan küreselleşme olgusu ülkeleri ve ekonomileri derinden etkiyen önemli bir gelişme olarak ortaya çıkmıştır. Ülkelerin küreselleşmenin etkilerine bağlı olarak ürettikleri politikalar, sergiledikleri yaklaşımlar ve belirlemeye çalıştıkları kararlılık çabaları ekonomi alanı başta olmak üzere birçok alanda yeni stratejiler ve eylemleri ortaya çıkarmıştır. Sınırların ortadan kalktığı, neoliberalleşmenin etkilerinin kendisini göstermeye başladığı bu süreçte ekonomilerinde sınırlarının kaldırması ve sermayenin sınırlar ötesinde hareketlilik göstermesi de yine küreselleşmenin bir çıktısı olarak görülmektedir. Buna bağlı olarak Türkiye ekonomisinde 1980 öncesindeki yaklaşım ile 24 Ocak kararları sonrasındaki yaklaşım azasında farklılıkların temelinde de yine bu sürecin etkileri ve izleri yer almaktadır (Öztürk ve Özyakışır, 2005). Küreselleşmenin etkilerine bağlı olarak ekonominin dışa açılması gibi atılımlar ve 24 Ocak kararlarıyla uygulamaya konulan neoliberal ekonomik politikalar, 1980’li yılların sonuna doğru tıkanma göstermiş ve ekonominin sürdürülebilirliği için yeni arayışlar içerisine girilmiştir. Ekonomik krizler, IMF ile imzalanan anlaşmalar, döviz kurunun çarpan aracı olarak kullanılma stratejisi, ekonomik istikrarın sağlanması için atılan birçok adım vb. girişimlerin yegane amacı Türkiye ekonomisini güçlü ve sürdürülebilir kılmaya yöneliktir (Cicioğlu, 2013: 30-42). Türkiye’deki 29 Bengü Kayapalı Aslan \| 29 Bengü Kayapalı Aslan ekonomik seyrin gelişmesine yönelik olarak bu gelişmeler aynı zamanda gelir dağılımı adaletini de etkilemektedir. Türkiye’de 1980 yılından günümüze kadar olan süreçte gelir dağılımı ölçüm yöntemlerinden Gini katsayısının almış olduğu değerler yıllık bazda incelenmiş olsa da iç ve dış kaynaklı olmak üzere ülkede yaşanan siyasi, toplumsal ve ekonomik birçok gelişme bu süreci derinden etkilemiştir. 5.Türkiye’de Gelir Dağılımının Seyri Devletler, ülkelerinde yaşayan insanların tamamına özgürlük ve güven, istikrar ve adalet sağlayan siyasal bir örgüttür. Buna bağlı olarak bu örgütün önemli bir misyonu da fırsat eşitliğini oluşturabilmek için gelir dağılımındaki adaleti de sağlamaktır. Aksi halde marjinal verimliliğe göre oluşan gelir dağılımı, kaynakları paylaşımı noktasında etkinliği sağlamaya muktedir olsa da fırsat eşitliğini tam anlamıyla sağlayamadığından ötürü gelir dağılımı noktasından eşitsizlikler meydana getirmektedir (Yüce, 2001: 1-2). Türkiye’de gelir dağılımına ilişkin görünüm için bu çalışmada 2006-2020 gözlem aralığını kapsayan yıllık TÜİK Gini katsayısı verilerden faydalanılmıştır (TÜİK, 2023). Tablo.1 Türkiye’de Kişisel Gelir Dağılımı (2006-2020) Kaynak: Türkiye İstatistik Kurumu (TÜİK), 2023. Tablo.1 Türkiye’de Kişisel Gelir Dağılımı (2006-2020) Kaynak: Türkiye İstatistik Kurumu (TÜİK), 2023. Gelir dağılımı eşitsizliğine ilişkin seyir, Türkiye İstatistik Kurumu’nun verilerine bağlı olarak yukarıdaki tabloda gösterilmiştir. Gelir dağılımı eşitsizliği ölçüm yöntemlerden yaygın olarak kullanılan Gini katsayı değerleri incelendiğinde öncelikle Türkiye’de 2006 ile 2020 yılları arasında gelir eşitsizliğinde hem artan hem de azalan bir seyir göstermektedir. Gini katsayılarının yüzdelik ve bindelik küsuratları her ne kadar tabloda verilmemiş olsa da katsayı 2006 ile 2009 yılları arasında azalan bir seyir gösterirken 2009 yılında artmıştır. 2010 ile 2014 yılları arasında Gini katsayısındaki azalmaya Türkiye’de Gelir Dağılımına İlişkin Görünüm 30 bağlı olarak gelirin bölüşünü noktasında görece eşitlikçi bir konuma doğru ilerlenmiştir. Ancak 2015 yılı sonrasında Gini katsayısındaki artışa bağlı olarak gelir eşitsizliğinde belirgin bir artış yaşanmış ve gelir dağılımındaki eşitsizlik durumu artma eğilimine girmiştir. Dünya’da yaşanan koronavirüs salgını sonrasında ülkelerin hem ekonomik hem de sosyal dinamiklerinin olumsuz şekilde etkilenmesine bağlı olarak gelir dağılımı tablosunda 2020 yılı sonrasındaki verilere yer verilmemiş olsa da Türkiye’nin son üç yıldaki verileri, Gini katsayısının artan seyri ile devam ettiğini de belirtmek gerekir. Sonuç olarak yukarıdaki tablodan hareketle Türkiye’de 2006-2020 yılları arasındaki dönem göstermektedir ki Türkiye gelir dağılımında eşitliğin sağlanması konusunda istikrarlı bir iyileşme sürecine sahip olmadığı görülmektedir. İncelenen bu seyir içerisinde nüfusun en düşük gelirli yüzdesinin almış olduğu payın son dönemlerde azalma eğilimi içerisinde olmasına karşın gelirden en yüksek payı alan kesime ilişkinin yüzdenin artış seyrinde olması gelirin dağılımında bir eşitsizlik durumunun olduğunu ayrıca göstermektedir. Buradan hareketle gelir dağılımında ek yüksek pay alanlar ile en düşük pay alanlar arasındaki oranın son dönem itibariyle artan değerler alması, gelir dağılımı eşitsizliğinin artan bir seyirde olduğunu ortaya çıkarmasının yanında buna paralel olarak yoksulluğunda artan bir seyir izlediğine ilişkin sonuçlar barındırmaktadır (Mayda ve Vurkun, 2018: 216) . Sonuç Bir toplumdaki gelir eşitsizliğini istatistiksel verilerle ortaya koyabilmek kolay olsa da adil bir gelir dağılımını sağlamak önemli derece zordur. Gelişmiş ve gelişmekte olan ülkelerin amaçlarından birisi de sürdürülebilir bir ekonomik büyümeyi sağlamalarının yanında gelirin adil dağılmasının sağlanmasıdır. Gelir dağılımındaki adaletin sağlanması, toplumdaki eşitsizliğin çözümünde önemli rol oynamaktadır. Gelir dağılımındaki eşitsizlik günümüz sosyoekonomik sorunlarının başında gelmektedir. Gelirin paylaşımındaki adaletsizlik, az gelişmiş ve gelişmekte olan ülkelerde gelişmiş ülkelere kıyasla görece daha fazla olduğu bilinmektedir. Ülkelerin siyasi, ekonomik ve toplumsal yapıları birbirlerinden farklılık gösterse de gelir dağılımındaki eşitsizlik durumu toplumdaki eşitliğin ve huzurun sağlanmasında ortak etkilere sahiptir. Gelir dağılımı adaletinin sağlanması hem ekonomik hem de toplumsal çıktılar ortaya çıkaracaktır. Gelir dağılımının sağlanması noktasındaki her boşluk veya politika etkisizliğinin toplumda birtakım sorunlar ve huzursuzluklar meydana getireceği de unutulmamalıdır. Bu sebeple ülkelerin gelir dağılımı eşitsizliğinin çözümüne ve gelir dağılımı adaletinin tesisine yönelik politikalar geliştirmesi ayrıca önem taşımaktadır. 31 Bengü Kayapalı Aslan \| Bengü Kayapalı Aslan \| Günümüzde gelirin adil olarak dağılımı ülkeler için tam anlamıyla gerçekleşmiş bir durumun ötesinde kısa veya uzun vadede belirlenmiş hedefler arasında yer alan bir amaç olarak yer almaktadır. Milli gelirin bölüşümünde adil olma motivasyonu yer alsa da bölgeler arasındaki farklılıklar başta olmak üzere ülkelerin kendilerine ait dinamikleri bu bölüşümü etkilemektedir. Bu bölüşümün ölçülmesinde birtakım yöntemler kullanılmaktadır. Çalışmada, gelir dağılımı ölçüm yöntemleri arasında yaygın olarak kullanılan bir yöntem olan Gini katsayısı verilerinden yararlanılarak birtakım sonuçlara ulaşılmıştır. Gelir dağılımı eşitsizliğinin ölçümünde yararlanılan Gini katsayısı, 0 ile 1 arasında değerler almaktadır. Gelir adaletsizliğinde artış olduğu anlamı taşıyan Gini katsayısının 1’e yaklaşması durumu ülkelerin karşılaşmak istemediği bir durumu oluşturmaktadır. Buna bağlı olarak Türkiye’nin gelir dağılımına ilişkin görünümüne değinmeyi amaçlayan bu çalışmada Türkiye İstatistik Kurumu (TUİK) Gini katsayısı verilerinden yararlanılarak belirli bulgulara ulaşılmıştır. Yıllar itibariyle incelenen Gini katsayısına ilişkin veriler, Türkiye’de gelir dağılımı eşitsizliğine ilişkin seyrin belirlenmesinde referans noktası olmuştur. Çalışmada oluşturulan Gini katsayısı verilerine ait tablo göstermektedir ki Türkiye’de gelir dağılımına ilişkin süreçte gelir dağılımı eşitsizliğinde zaman zaman artış görüldüğü gibi azalışlar da görülmüştür. Buna bağlı olarak Türkiye gelir dağılımında eşitliğin sağlanması konusunda istikrarlı bir iyileşme sürecine sahip olmadığı görülmüştür. Bu seyrin istikrarlı bir şekilde devam etmemesinde Türkiye’de yaşanan siyasi, ekonomik, toplumsal ve uluslararası gelişmelerin etkisi olmuştur. Sonuç olarak Türkiye’de gelir dağılımına ilişkin görünümün incelendiği bu çalışmada Gini katsayısının 0’a yaklaştığı dönemler ve 1’e yaklaştığı dönemlerin yer aldığı görülmüştür. Sonuç Bu dönemler itibariyle Türkiye’de yaşanan gelişmeler bu seyri etkilese de gelir dağılımındaki eşitsizliğin giderilmesine yönelik politikaların üretilmesi gerek ülkenin refahı gerekse toplumun huzuru için önem taşımaktadır. Gelir dağılımındaki eşitsizlik sorununun çözümüne ilişkin olarak atılan ve atılacak adımlar ayrıca sürdürülebilir kalkınma, yoksulluk sorunu, fırsat eşitliği vb. gibi birçok durumun sağlanmasında pay sahibi olacaktır. 32 \| Türkiye’de Gelir Dağılımına İlişkin Görünüm 32 34 \| Türkiye’de Gelir Dağılımına İlişkin Görünüm 2 Dr. Öğr. Üyesi, Şırnak Üniversitesi, Silopi Meslek Yüksekokulu, odemir@sirnak.edu.tr, 0000-0001-8421-0619. 1 Dr. Öğr. Üyesi, Şırnak Üniversitesi, İktisadi ve İdari Bilimler Fakültesi, snas@sirnak.edu.tr, 0000-0003-3267-4432. 1 Dr. Öğr. Üyesi, Şırnak Üniversitesi, İktisadi ve İdari Bilimler Fakültesi, snas@sirnak.edu.tr, 0000-0003-3267-4432. 2 Dr. Öğr. Üyesi, Şırnak Üniversitesi, Silopi Meslek Yüksekokulu, odemir@sirnak.edu.tr, 0000-0001-8421-0619. Kaynakça Aksoy, Ş. (1989). Vergi Hukuku ve Türk Vergi Sistemi, Filiz Kitabevi, İstanbul. Asandaş, N., & Işık, N. (2021). Ekonomik Kalkınma ve Gelir Dağılımının Yok- sulluk Üzerindeki Etkisi: Ekonometrik Bir Analiz. Aksaray Üniversitesi İktisadi ve İdari Bilimler Fakültesi Dergisi, 13(3), 101-116. https://doi. org/10.52791/aksarayiibd.876865 Cicioğlu, Ş. (2013). Türkiye’de 1980 Sonrası Gelir Dağılımının Ekonomi Po- litikaları ve Krizler Bağlamında Değerlendirilmesi. Hak İş Uluslararası Emek ve Toplum Dergisi, 2(4), 28-49. Clements, B., Gaspar, V., Gupta, S., & Kinda, T. (2015). “Chapter 2. The IMF and Income Distribution”. In Inequality and Fiscal Policy. USA: In- ternational Monetary Fund. Retrieved Dec 6, 2023, from https://doi. org/10.5089/9781513567754.071.ch002 Çalışkan, Ş. (2010). Türkiye’de Gelir Eşitsizliği ve Yoksulluk. Journal of Social Policy Conferences (59), 89-132. Çavuşoğlu, T. ve Hamurdan, Y. (1966). Gelir Dağılımı Araştırması. Ankara: DPT Yayınları. Dinçer, M. R., ve Kalelioğlu, M. H. (2013). Gelirin Tespiti ve Vergilendirilmesi: Safi Artış Teorisi, Kaynak Teorisi. Ufuk Üniversitesi Sosyal Bilimler Enstitü- sü Dergisi, 2(3), 85-104. Işık, N. (2006). Sektörel Gelir Dağılımının Ekonomik Büyüme Üzerindeki Et- kisi. Karamanoğlu Mehmetbey Üniversitesi Sosyal ve Ekonomik Araştırmalar Dergisi, 2006(2), 119-127. İlhan, A. (2015). Türkiye’de Gelir Dağılımı, Hak İş Uluslararası Emek ve Toplum Dergisi, 4(8), 42-59 Karataş, M. (2019). Gelir Dağılımının Teorik Yapısı. Hak İş Uluslarara- sı Emek ve Toplum Dergisi, 8(21), 52-79. https://doi.org/10.31199/ hakisderg.546147 MAYDA, F. A., ve VURKUN, S. (2018). Gelir Dağılımı Eşitsizliğinin Toplum- sal Cinsiyet Bağlamında Değerlendirilmesi. The Journal of International Lingual Social and Educational Sciences, 4(2), 214-228. Öztürk S. ve Özyakışır, D. (2005). Türkiye Ekonomisinde 1980 Sonrası Yaşa- nan Yapısal Dönüşümlerin Gsmh, Dış Ticaret ve Dış Borçlar Bağlamında Teorik Bir Değerlendirmesi, Mevzuat Dergisi, 94. Pehlivan, M. S. (2009). Gelir Dağılımı Eşitsizliğine Devletin Müdahale Araçla- rı: Sosyal Yardımlaşma ve Dayanışmayı Teşvik Fonu Örneği, T.C. Başba- kanlık Sosyal Yardımlaşma ve Dayanışma Genel Müdürlüğü Sosyal Yar- dım Uzmanlık Tezi, Ankara. 33 Bengü Kayapalı Aslan Sahota, G. S. (1978). Theories of Personal Income Distribution: A Sur- vey. Journal of Economic Literature, 16(1), 1–55. http://www.jstor.org/ stable/272312 Sefil Tansever, S., & Kent, O. (2018). Earnıngs Inequality In Turkey: A Regio- nal Perspective. Marmara İktisat Dergisi, 2(1), 117-136. Şerbetçi, G. (2015). Türkiye’de Uygulanan Finansal Politikaların Gelir Dağılı- mı Üzerindeki Etkisi. Kahramanmaraş Sütçü İmam Üniversitesi İktisadi ve İdari Bilimler Fakültesi Dergisi, 4(1), 93-116. Türkiye İstatistik Kurumu (2023). Gelir Dağılımı İstatistikleri, 2022, ht- tps://data.tuik.gov.tr/Bulten/Index?p=Gelir-Dagilimi-Istatistikle- ri-2022-49745, Erişim Tarihi: 04.12.2023 UNDP (2015). What are the Sustainable Development Goals?, https://www. Kaynakça undp.org/sustainable-development-goals?gad_source=1&gclid=CjwK- CAiAjrarBhAWEiwA2qWdCAYmfhh8vu7gs-b_7df_nse1NijJvDRw- FuLcdX853CZPUUgeEOyRiRoCvb4QAvD_BwE, Erişim Tarihi: 29.11.2023 Bölüm 3 Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik Büyüme: BRICS ve MINT Ülkeleri Üzerine Ampirik Bir İnceleme Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik Büyüme: BRICS ve MINT Ülkeleri Üzerine Ampirik Bir İnceleme Şahin Nas1 Ömer Demir2 Özet Bir ekonomide büyümenin birçok belirleyicisi ve dinamiği bulunmaktadır. Bu anlamda bilgi ve iletişim teknolojileri (BİT) ve yenilenebilir enerji tüketimi (YET) gibi faktörler de büyüme için önemli dinamiklerdir. Çünkü 1980 sonrasında başlayan küreselleşme eğilimlerinin en önemli itici gücü BİT olmuştur ve küreselleşme eğilimlerini hızlandırmıştır. Aynı şekilde BİT’ler toplam faktör verimliliğini artırarak dünya ekonomisinde meydana gelen küresel üretim artışının önemli bir etkeni olmuştur. Bununla birlikte küresel üretimin artış göstermesi ve farklı coğrafyalara yayılmasıyla birlikte mal ve hizmet üretiminde yenilenebilir enerji tüketimi de temel girdilerden biri haline gelmiştir. Bu çerçevede çalışmanın temel amacı, BRICS (Brezilya, Rusya, Hindistan, Çin ve Güney Afrika) ve MINT (Meksika, Endonezya, Nijerya ve Türkiye) ülkelerinde BİT ve YET’nin büyümeyi nasıl ve ne derecede etkilediğini analiz etmektir. Bu bağlamda, bu ülkelerin 1995-2018 dönemine ait verileri kullanılarak Panel ARDL/PMG yöntemi uygulanmıştır. Analiz sonucunda elde edilen bulgulara göre, BİT ve YET uzun dönemde ekonomik büyümeyi pozitif etkilemektedir. Elde edilen bulgular çerçevesinde, BRICS ve MINT ülkelerinde sürdürülebilir ve kalıcı bir büyüme için BİT ve YET alanlarında seçici politikalar üretilmeli ve bu alanlara sürekli yatırım yapılmalıdır. https://doi.org/10.58830/ozgur.pub401.c1751 https://doi.org/10.58830/ozgur.pub401.c1751 35 36 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... 1. Giriş Bilgi ve iletişim teknolojileri (BİT) sektörü, 1940’lı yıllarda mikro elektriğin icadı, 1960’lı yıllarda bilgisayar teknolojisinin yavaş yavaş gelişmesi, 1990’lı yıllarda internetin kullanılması ve yaygınlaşması ve son dönemlerde yapay zekâ ve blok zincir teknolojisinin gelişmesine kadar son yıllarda hızlı ve radikal bir gelişme yaşamıştır (Bakry vd., 2023). Bu bağlamda, 21.yüzyılda teknolojik gelişmeler özellikle BİT’ler dünyada hızlı bir şekilde yayılmıştır. Teknolojik gelişmelerin hızlanması ve yayılması üretim sürecini hızlandırmakta ve maliyet avantajları sağlamaktadır. Özellikle, BİT’lerin üretim sürecini hızlandırması ve küresel mekanizmalar üzerinde önemli bir etkisi olduğu söylenebilir. Bununla birlikte bir ekonomide BİT, sürdürülebilir ekonomik büyüme, refah ve kalkınma için önemli bir dinamik olduğu ileri sürülebilir. Çünkü, bu teknolojiler, maliyet avantajı sağlayarak verimliliği artırmakta ve rekabet avantajı sağlamaktadır (Tsimisaraka vd., 2023; Verma, Dangawhal & Giri, 2023). BİT’ler, süreçlerin, kontrollerin ve bilgi üretiminin otomasyonuna ek olarak bilginin depolanması, geri alınması, dönüştürülmesi ve iletilmesini içermektedir. Bu açıdan BİT’ler, bir toplumda hızlı değişmeler yaratan ve küresel ekonominin şekillenmesinde önemli bir faktördür. Ayrıca BİT’ler son dönemlerde iletişim kalıplarını ve iş ilişkilerini radikal bir şekilde değiştirmiştir. BİT’ler ülkelerin kalkınmaları ve kapasitelerini geliştirmeleri için gerekli geleneksel aşamaları veya süreçleri atlamasına olanak tanımaktadır. Bu durum da bir ülkenin teknoloji sıçraması olarak tanımlanmaktadır (Adeleye & Eboagu, 2019). Analize dahil edilen BRICS (Brezilya, Rusya, Hindistan, Çin, Güney Afrika) ve MINT (Meksika, Endonezya, Nijerya, Türkiye) ülkelerinde genel anlamda BİT’lerin ekonomideki durumunu görebilmek amacıyla Şekil 1 verilmiştir. Şekil 1: BİT ürünlerinin Toplam Ticaret İçindeki Payı, 2000-2020 (%) Kaynak: UNCTAD (2023) Şekil 1: BİT ürünlerinin Toplam Ticaret İçindeki Payı, 2000-2020 (%) Kaynak: UNCTAD (2023) Şahin Nas / Ömer Demir \| 37 37 Şahin Nas / Ömer Demir Şekil 1’de BRICS ve MINT ülkelerinde BİT ürünlerinin toplam ticaret içindeki payı verilmiştir. Şekil 1’de de görüldüğü üzere Çin’de BİT’in toplam ticaret içindeki payı diğer ülkelere göre daha yüksektir. Çin’de BİT ürünlerinin toplam ticaret içindeki payı nerdeyse 2006 yılına kadar sürekli artış göstermiştir. Ancak daha sonra düşüş gösterse de genel olarak dönem sonuna kadar stabil bir seyir izlediği söylenebilir. Çin’de BİT ürünlerinin toplam ticaret içindeki payı 2000-2020 döneminde ortalama %26,90’dır. Çin’den sonra Meksika’da BİT ürünlerinin toplam ticaret içindeki payı en yüksektir. Meksika’da BİT ürünlerinin toplam ticaret içindeki payı genel anlamda düşüş eğiliminde olduğu söylenebilir. 2000-2020 döneminde Meksika’da BİT ürünlerinin toplam ticaret içindeki payı ortalama %18’dir. Endonezya’da dönem başında BİT ürünlerinin toplam ticaret içindeki payı %12 civarında iken, bu oran dönem boyunca düşüş göstermiş ve dönem sonunda BİT ürünlerinin toplam ticaret içindeki payı yaklaşık %3’tür. 1. Giriş Şekil 1’de görüldüğü üzere diğer ülkelerde BİT ürünlerinin toplam ticaret içindeki payı çok düşük ve bu oran %0-2 arasında değişkenlik göstermektedir. BİT gibi son dönemlerde yenilenebilir enerjinin de üretimde önemli bir girdi bileşenini oluşturduğu ve ekonomik büyüme için önemli bir dinamik olduğu ileri sürülebilir. Teknolojik gelişmeler, nüfusun artması, kentleşmenin hızlanması, sınai gelişme gibi faktörler enerji talebini de artırmaktadır (Yang vd., 2022). Bu bağlamda enerjinin büyümeye etkisi dikkat çekmektedir (Chen vd., 2023). Ancak bununla birlikte artan enerji tüketimi (özellikle büyük miktardaki kirletici gaz emisyonları) da insan sağlığını, ekonomik büyümeyi ve sosyal düzeni ciddi bir şekilde olumsuz etkilemektedir. Ayrıca küresel ısınma, deniz seviyesinin yükselmesi, hava kirliliği gibi problemler de ortaya çıkmaktadır. Bu nedenle bu tür problemlerin önüne geçebilmek ve ayrıca sürdürülebilir ekonomik büyüme ve kalkınma için yenilenebilir enerji kaynaklarının tüketimi önem arz etmektedir. Ayrıca yenilenebilir enerji kaynakları, endüstriyel alt yapının iyileştirilmesini teşvik edebilir ve diğer enerji kaynaklarına göre daha fazla istihdam olanağı sağlayabilir. Dolayısıyla yenilenebilir enerji tüketimi sürdürülebilir ekonomik büyümeyi destekleyebilir ve istihdamı artırabilir (Li vd., 2022; Tsimisaraka vd., 2023). Şekil 2, BRICS ve MINT ülkelerinde yenilenebilir enerji tüketiminin toplam enerji tüketimi içindeki payını göstermektedir. 8 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... 38 Şekil 2: Yenilenebilir Enerji Tüketimi, 2000-2020 (%) Kaynak: WDI (2023) Şekil 2: Yenilenebilir Enerji Tüketimi, 2000-2020 (%) Kaynak: WDI (2023) Şekil 2’ye göre 2005 yılından sonra Brezilya’da yenilenebilir enerji tüketiminin toplam enerji tüketimi içindeki payı diğer ülkelere göre daha yüksektir. Brezilya’da yenilenebilir enerji tüketiminin toplam enerji tüketimi içindeki payı ortalama %45’tir. Brezilya’dan sonra enerji tüketiminin toplam nihai enerji tüketimi içindeki payı en yüksek olan ülkeler Hindistan ve Endonezya’dır. Ancak bu ülkelerde enerji tüketiminin düşüş eğiliminde olduğu görülmektedir. Çin ekonomisinde dönem başında yenilenebilir enerji tüketimi oranı yaklaşık %30 iken dönem sonunda bu oran yaklaşık %15’e düşmüştür. Türkiye ekonomisinde yenilenebilir enerji tüketimi dönem boyunca ortalama %14 civarındadır. Yenilenebilir enerji tüketiminin en düşük olduğu ülke ise Rusya’dır. Güney Afrika ve Meksika’da da yenilenebilir enerji tüketiminin düşüş eğiliminde olduğu söylenebilir. Şahin Nas / Ömer Demir 39 Şekil 3: Ekonomik Büyüme Oranları, 2000-2020 (%) Kaynak: WDI (2023) Şekil 3: Ekonomik Büyüme Oranları, 2000-2020 (%) Kaynak: WDI (2023) Şekil 3, BRICS ve MINT ülkelerinde ekonomik büyüme oranlarını göstermektedir. 2010 yılına kadar neredeyse en yüksek ekonomik büyüme oranı Çin ekonomisinde gerçekleştiği söylenebilir. Çin’de 2000-2020 döneminde ortalama büyüme oranı %8,69’dur. En düşük ekonomik büyüme oranı ise Meksika ekonomisinde gerçekleşmiştir. Meksika’da 2000- 2020 döneminde ortalama ekonomik büyüme oranı %1,59’dur. 1. Giriş Diğer ülkelerde ise 2000-2020 döneminde ortalama büyüme oranı Brezilya’da %2,15, Rusya’da %3,45, Hindistan’da %5,88, Güney Afrika’da %2,24, Endonezya’da %4,91 ve Türkiye’de %4,79’dur. Özellikle 2008-2009 küresel finans krizi döneminde bütün ülkelerde ekonomik büyüme ciddi anlamda düşüş göstermiştir. BRICS ve MINT ülkelerin ekonomik büyüme verileri değerlendirildiğine ekonomik büyümenin dalgalı bir seyir izlediği ve özellikle kriz dönemlerinde büyümenin olumsuz etkilendiği görülmektedir. Dolayısıyla bu çalışmanın temel motivasyonu da BRICS ve MINT ülkelerinde ekonomik büyümenin sürdürülebilirliğini sağlamada bilgi ve iletişim teknolojileri (BİT) ve yenilenebilir enerji tüketiminin (YET) rolünü incelemektir. Bu amaçla BİT ve yenilenebilir enerjinin ekonomik büyüme için önemi ampirik olarak analiz edilmiştir. Ancak ampirik analiz kısmında Ekonomik Kalkınma ve İşbirliği Örgütü (OECD) veri tabanında Nijerya ekonomisine ait BİT verileri olmadığından ötürü Nijerya analizlere dahil edilmemiştir. Çalışma dört kısımdan oluşmaktadır. Birinci kısımda giriş yer almaktadır. Giriş kısmında genel olarak BİT ve yenilenebilir enerjinin önemi ve analize dahil edilen ülkelere ait veriler incelenmiştir. İkinci kısımda konu ile ilgili literatür 40 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... 40 taraması yer almaktadır. Üçüncü bölümde çalışmanın ampirik kısmı yer almaktadır. Son olarak dördüncü bölümde sonuç kısmı yer almaktadır. 2. Literatür İncelemesi Bu çerçevede Dees & Auktor (2018) de MENA ülkeleri için yenilenebilir enerji kaynaklarının yaygınlaşmasının büyüme için olumlu etkiler yaratığını vurgulamaktadır. Yang, Zhou & Feng (2022), Gelecek 11 ülkelerinde (Bangladeş, Endonezya, Filipinler, Güney Kore, İran, Meksika, Mısır, Nijerya, Pakistan, Türkiye ve Vietnam) 1990-2020 döneminde sadece kısa dönemde yenilenebilir enerji tüketiminin ekonomik büyümeyi hızlandırdığını ileri sürmektedir. Ahmad & Majeed (2022) de 5 Güney Asya ülkesi için yaptıkları analizde, 1990- 2018 döneminde yenilenebilir enerji tüketiminin çıktı büyümesi üzerinde pozitif ve anlamlı etkisi olduğu belirtilmektedir. Peçe vd. (2023) ise 1965- 2019 dönemi için Türkiye ekonomisine yönelik yaptıkları çalışmada, uzun ve kısa dönemde yenilenebilir enerjinin ekonomik büyüme üzerinde anlamlı bir etkisi olmadığını ileri sürmektedir. Bu anlamda Peçe vd. (2023), Türkiye ekonomisine yönelik elde etikleri bulgular Ocal & Aslan (2013) çalışmasında elde edilen bulgulardan farklılaşmaktadır. Bunun nedeni ise her iki çalışmada ele alınan dönemin farklı olmasından kaynaklı olduğu söylenebilir. Madaleno & Nogueira (2023), 1994-2019 döneminde 27 Avrupa ülkesinde, Jia, Fan & Xia (2023) 2000-2019 döneminde Kuşak ve Yol üzerindeki ülkelerinde, Shen & Ren (2023) Çin’de yenilenebilir enerji tüketiminin büyümeyi pozitif ve anlamlı etkilediğini belirtmektedir. Ancak Aswadi vd. (2023) tarafında yapılan çalışmada, 1990-2019 döneminde Endonezya’da yenilenebilir enerji tüketiminin büyümeyi negatif etkilediği ileri sürülmektedir. Gelişmekte Olan Yedi Ülke (E-7)’de 1992-2012 döneminde yenilenebilir enerji tüketiminin reel GSYH’yi pozitif etkilediğini belirtmektedir. Benzer şekilde Koçak & Şarkgüneşi (2017), 1990-2012 döneminde Karadeniz ve Balkan ülkelerinde de uzun dönemde yenilenebilir enerji tüketimi ve ekonomik büyüme arasında bir ilişki olduğunu ve aynı zamanda yenilenebilir enerjinin büyümeyi pozitif etkilediğini ileri sürmektedir. Bu çerçevede Dees & Auktor (2018) de MENA ülkeleri için yenilenebilir enerji kaynaklarının yaygınlaşmasının büyüme için olumlu etkiler yaratığını vurgulamaktadır. Yang, Zhou & Feng (2022), Gelecek 11 ülkelerinde (Bangladeş, Endonezya, Filipinler, Güney Kore, İran, Meksika, Mısır, Nijerya, Pakistan, Türkiye ve Vietnam) 1990-2020 döneminde sadece kısa dönemde yenilenebilir enerji tüketiminin ekonomik büyümeyi hızlandırdığını ileri sürmektedir. Ahmad & Majeed (2022) de 5 Güney Asya ülkesi için yaptıkları analizde, 1990- 2018 döneminde yenilenebilir enerji tüketiminin çıktı büyümesi üzerinde pozitif ve anlamlı etkisi olduğu belirtilmektedir. Peçe vd. (2023) ise 1965- 2019 dönemi için Türkiye ekonomisine yönelik yaptıkları çalışmada, uzun ve kısa dönemde yenilenebilir enerjinin ekonomik büyüme üzerinde anlamlı bir etkisi olmadığını ileri sürmektedir. Bu anlamda Peçe vd. (2023), Türkiye ekonomisine yönelik elde etikleri bulgular Ocal & Aslan (2013) çalışmasında elde edilen bulgulardan farklılaşmaktadır. Bunun nedeni ise her iki çalışmada ele alınan dönemin farklı olmasından kaynaklı olduğu söylenebilir. 2. Literatür İncelemesi Çalışmada, BRICS ve MINT ülkelerinde BİT ve yenilenebilir enerjinin ekonomik büyüme için önemi analiz edilmiştir. 1980 sonrası dönem başta olmak üzere özellikle 2000’li yıllardan sonra ekonomik açıdan BİT önemli bir dinamik haline geldiği söylenebilir. Küreselleşme eğilimlerinin hızlanması, dünya ekonomisinde üretimin parçalanması BİT’lerin ekonomik önemini artırmıştır. Benzer şekilde son dönemlerde yenilenebilir enerjinin ekonomik önemi gittikçe artmış ve büyümeye katkısı yoğun bir ilgi gördüğü ileri sürülebilir. Bu bağlamda literatürde BİT ve yenilenebilir enerjinin büyüme için önemini yoğun bir şekilde analiz eden ciddi bir ampirik literatür mevcuttur. Bu amaçla çalışmanın bu kısmında BİT ve yenilenebilir enerji ile ilgili yapılan ampirik çalışmaların kısa bir literatür incelenmesi yapılmıştır. Apergis & Payne (2010) 1985-2005 dönemi için 20 OECD ülkesine yönelik yaptıkları analizde, uzun dönemde yenilenebilir enerji tüketimi ile büyüme arasında bir ilişki olduğunu ve bu dönemde yenilenebilir enerji tüketiminin büyümeyi pozitif etkilediği ileri sürmektedir. Çalışmada ayrıca hem kısa dönemde hem de uzun dönemde büyüme ve yenilenebilir enerji arasında çift yönlü bir nedensellik ilişkisi olduğuna dair bulgular sunulmaktadır. Ocal & Aslan (2013) ise Türkiye ekonomisine yönelik yaptıkları analizde, 1990-2010 döneminde yenilenebilir enerjinin büyümeyi negatif etkilediği belirtilmektedir. Pao & Fu (2013) Brezilya ekonomisinde, 1980-2010 döneminde yenilenemeyen enerji tüketimi ve toplam birincil enerji tüketiminin reel çıktı üzerinde anlamlı bir etkisi olmadığını ancak aynı dönemde hidrolik olmayan yenilenebilir enerji tüketiminin reel çıktıyı pozitif etkilediğini ileri sürmektedir. Lin & Moubarak (2014) 1977-2011 dönemi için Çin ekonomisine yönelik yaptıkları çalışmada, Apergis & Payne (2010) tarafından yapılan çalışmaya benzer bulgular sunmaktadır. Lin ve Moubarak (2014) Çin ekonomisinde yenilenebilir enerji tüketimi ve ekonomik büyüme arasında çift yönlü bir nedensellik ilişkisi olduğunu vurgulamaktadır. Cho, Eunnyeong & Kim (2015) 31 OECD ülkesi ve OECD üyesi olmayan 49 ülke için yaptıkları analizde, 1990-2010 döneminde gelişmiş ülkelerde yenilenebilir enerjinin ekonomik büyümede önemli bir rol oynamadığı belirtilmektedir. Bu açıdan çalışma, Apergis & Payne (2010) tarafından yapılan çalışma ile ayrışmaktadır. Ancak gelişmekte olan ülkelerde ise aynı dönemde, yenilenebilir enerjinin ekonomik büyümenin artmasına yol açtığını ve yenilenebilir enerjinin üretim girdisi için önemli bir rol oynadığı belirtilmektedir. Çetin (2016), Şahin Nas / Ömer Demir \| 41 41 Gelişmekte Olan Yedi Ülke (E-7)’de 1992-2012 döneminde yenilenebilir enerji tüketiminin reel GSYH’yi pozitif etkilediğini belirtmektedir. Benzer şekilde Koçak & Şarkgüneşi (2017), 1990-2012 döneminde Karadeniz ve Balkan ülkelerinde de uzun dönemde yenilenebilir enerji tüketimi ve ekonomik büyüme arasında bir ilişki olduğunu ve aynı zamanda yenilenebilir enerjinin büyümeyi pozitif etkilediğini ileri sürmektedir. 2. Literatür İncelemesi Madaleno & Nogueira (2023), 1994-2019 döneminde 27 Avrupa ülkesinde, Jia, Fan & Xia (2023) 2000-2019 döneminde Kuşak ve Yol üzerindeki ülkelerinde, Shen & Ren (2023) Çin’de yenilenebilir enerji tüketiminin büyümeyi pozitif ve anlamlı etkilediğini belirtmektedir. Ancak Aswadi vd. (2023) tarafında yapılan çalışmada, 1990-2019 döneminde Endonezya’da yenilenebilir enerji tüketiminin büyümeyi negatif etkilediği ileri sürülmektedir. Sassi & Goaied (2013), 17 MENA ülkesine yönelik yaptıkları çalışmada, 1960-2009 döneminde BİT bileşenlerinin ekonomik büyümeyi pozitif ve anlamlı bir şekilde etkilediğine dair bulgular sunulmaktadır. Lee & Brahmasrene (2014), 1991-2009 döneminde dokuz Güneydoğu Asya ülkesinde BİT’lerin ekonomik büyümeyi pozitif etkilediğini belirtmektedir. Saidi, Hassen & Hammami (2015)’ye göre Tunus ekonomisinde de BİT ekonomik büyümeyi pozitif etkilemektedir. Albiman & Sulong (2016) tarafında 45 Sahra-Altı Afrika ülkesi için bir analiz yapılmıştır. Analizde ele edilen bulgulara göre, 1999-2014 döneminde mobil telefon ve internet kullanımı ekonomik büyümeyi artırmaktadır. Ancak çalışmada, doğrusal olmayan etki analizi sonuçlarına göre, BİT bileşenleri ekonomik büyümeyi yavaşlattığı ileri sürülmektedir. Albiman & Sulong (2016)’ya benzer bir şekilde Adeleye & Eboagu (2019) de 54 Afrika ülkesinde 2005-2015 döneminde 42 \| 42 Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... BİT gelişiminin ekonomik büyümeyi pozitif etkilediğini belirtmektedir. Pradhan, Mallik & Bagchi (2018), 2001-2012 dönemi için G-20 ülkelerinde yönelik yaptıkları çalışmada, sabit geniş bant ve internet kullanımının kişi başına düşen GSYİH’yı artırdığına dair bulgular sunmaktadır. Pradhan vd. (2020) da 1989-2016 döneminde 25 Avrupa ülkesinde BİT yayılımının ekonomik büyümeye olumlu bir etki yarattığını ileri sürmektedir. Farklı gelir gruplarında yer alan 128 ülke için analiz yapan Appiah-Otoo & Song (2021) da genel anlamda 2002-2017 döneminde BİT’lerin ekonomik büyümeyi artırdığını vurgulamaktadır. Fakat Cheng, Chien & Lee (2021) 72 ülke için yaptığı analizde, orta ve düşük gelirli ülkelerde BİT’lerin ekonomik büyümeye etkisinin belirsiz olduğunu ancak yüksek gelirli ülkelerde ise BİT yayılımının ekonomik büyümeyi artırdığını ileri sürmektedir. Kurnawati (2020) 1996-2017 döneminde OECD ülkelerinde BİT yayılımının, Liu vd. (2021) 1990-2015 döneminde Pakistan’da internet penetrasyonu ve mobil telefon kullanımın, Singa & Sengupta (2022) 2001-2017 döneminde 30 Asya-Pasifik ülkesinde, Verma, Dandgawhal & Giri (2023) ise, 2005-2019 döneminde 88 gelişmekte olan ülkede BİT’lerin ekonomik büyümeyi pozitif etkilediğini ileri sürmektedirler. Literatürde hem yenilenebilir enerji ve hem de BİT’lerin ekonomik büyümeye etkisini birlikte analiz eden çalışmalar da yer almaktadır. Bu bağlamda Usman vd. (2021) 1990-2018 dönemi için Güney Asya ülkeleri (Bangladeş, Hindistan, Pakistan ve Sri Lanka) için yaptıkları analizde, sadece Hindistan’da uzun dönemde BİT’lerin ekonomik büyümeye etkisi pozitif ve anlamlı olduğu tespit edilmiştir. 2. Literatür İncelemesi Bununla birlikte çalışmada, enerji tüketiminin Hindistan ve Pakistan’da kişi başına düşen GSYH’nın önemli bir belirleyici olduğu vurgulanmaktadır. Zarkovic vd. (2022) ise AB üyesi ülkeler için yaptıkları çalışmada, 2000-2020 döneminde yenilenebilir enerji tüketimi ve BİT kişi başına düşen GSYH’yı pozitif etkilediğini ancak yenilenemeyen enerji tüketiminin ise kişi başına düşen GSYH’yı negatif etkilediğine yönelik bulgular sunmaktadır. Dehmani, Mabrouki & Youssef (2023) ise 1980-2018 dönemine MENA ülkelerinde yenilenebilir enerji tüketimi, yenilenemeyen enerji tüketimi ve BİT’lerin ekonomik büyümeyi pozitif ve anlamlı etkilediğini belirtmektedir. 3. Veri ve Yöntem Bu çalışmanın metodolojik çerçevesi; veri ve modelin tanımlanması, teorik ve ampirik spesifikasyondan oluşmaktadır. 43 Şahin Nas / Ömer Demir \| 3.1.Veri ve Model Bu çalışma, 1995-2018 döneminde BRICS ve MINT ülkelerinde bilgi ve iletişim teknolojileri (BİT) ile yenilenebilir enerji tüketiminin (YET) çalışan başına GSYH üzerindeki etkisini ampirik olarak araştırmaktadır. Bu ilişkiyi tahmin etmek için aşağıdaki teorik modelden yararlanılmıştır: (1) Eşitlik (1)’de GSYH, 2017 satın alma gücü paritesi ABD doları cinsinden ölçülen çalışan başına reel gayri safi yurtiçi hasılayı (GSYH); BİT, toplam üretimin % katma değeri, YET, toplam nihai enerji tüketimi % olarak yenilenebilir enerji tüketimini temsil edilmektedir. Çalışan başına GSYH verisi Penn World Table (PWT 10.0) veri tabanından alınarak logaritmik dönüştürme işlemi uygulandıktan sonra farkı alınarak büyüme oranı elde edilmiştir. BİT verisi OECD veri tabanı; YET verisi ise Dünya Bankası (WB) tabanından elde edilmiştir. Tablo 1’de çalışmada kullanılan veri seti ve veri kaynakları özetlenmektedir. Tablo 1: Değişkenler ve Veri Kaynakları Değişken Tanım Ölçüm Kaynak lnGSYH Çalışan başına reel GSYH Milyon 2017 satın alma gücü paritesi cinsinden ABD doları PWT (2023) lnBİT Bilgi ve iletişim teknolojileri Toplam üretimin % olarak katma değeri OECD (2023) lnYET Yenilenebilir enerji tüketimi % Toplam nihai enerji tüketimi WB (2023) Tablo 1: Değişkenler ve Veri Kaynakları Eşitlik [1]’deki teorik modelimizin doğal logaritmik formu şöyledir. (2) (2) Eşitlik [2]’de, katsayıları, i dönemi, t yatay kesiti ve hata terimini ifade etmektedir. Böylece modelimizin tam logaritmik formu tahmin edilecektir. 3.2. Ampirik Strateji Bu çalışmada izlenen ampirik strateji birkaç adımdan olumaktadır. Birinci adımda, yatay kesit bağımlılığı test edilmektedir. İkinci adımda panel durağanlık sınaması yapılmıştır. Üçüncü adımda uzun dönem katsayıları tahmin edilmiştir. Dördüncü adımda ise panel nedensellik testi uygulanmıştır. 44 4 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... Birinci adım, paneli oluşturan seriler arasında yatay kesit bağımlığının olup olmadığını sınamaktır. Yatay kesit bağımlılığının sınanmasında Breusch-Pagan (1980) LM testi, Pesaran (2004) CD testi ve Pesaran, Ullah & Yamagata (2008) tarafından geliştirilen Bias-corrected scaled LM testi kullanılmaktadır. Yatay kesit boyutunun zaman boyutundan büyük (N>T) olduğu durumlarda Breusch-Pagan (1980) LM testi kullanılamamakta iken, Pesaran (2004) ve Pesaran Ullah & Yamagata (2008) CD testleri kullanılabilmektedir. İkinci adım olarak, bu çalışmada Pesaran’ın CIPS panel birim kök testi kullanılmıştır. CIPS birim kök testinin faydası, yatay kesitler arasındaki yatay kesit ilişkilerini hesaba katmasıdır. Üçüncü adımda, modelin uzun dönem esneklerini tahmin etmek için Pesaran Shin & Smith (1999)’dan hareketle ampirik formda panel ARDL (Gecikmesi dağıtılmış otoregresif model) biçiminde şu şekilde ifade edilebilir: (3) Eşitlik (3), kısa ve uzun dönem dinamikleri olmak üzere iki kısımdan oluşmaktadır. Burada uzun dönem katsayıları 1 β , 2 β ve 3 β ; kısa dönem katsayıları ise iλ , 1δ ve 2 δ sembolleriyle ifade edilmiştir. Eşitlikte yer alan sembollerden p ve q, sırasıyla bağımlı değişken ve dağıtılmış gecikme kısmında kalan değişken için ilgili gecikme sıralarını; ise hata terimini göstermektedir. Panel ARDL modelinde PMG tahmincisi kullanılmıştır. Dördüncü adım olarak, bilgi ve iletişim teknolojileri ve yenilenebilir enerji tüketimi ile ekonomik büyüme arasındaki uzun vadeli nedensellik ilişkilerini ortaya koymak için Dumitrescu-Hurlin (D-H) panel nedensellik testi uygulanmıştır. Panelde nedensellik olmadığını ifade eden sıfır hipotezinin aksine, D-H testinin alternatif hipotezi en az bir yatay kesit biriminde nedensel bir ilişki olduğunu göstermektedir (Dumitrescu & Hurlin, 2012). 4. Bulgular ve Tartışma Bu çalışmada BRICS ve MINT ülkelerinde BİT ile YET’in çalışan başına GSYH arasındaki ilişki 1995-2018 dönemi kapsamında panel ARDL / PMG yöntemi kullanılarak tahmin edilmiştir. Şahin Nas / Ömer Demir \| 45 Tablo 2: Tanımlayıcı İstatistikler Tablo 2: Tanımlayıcı İstatistikler Tablo 2: Tanımlayıcı İstatistikler Değişken Gözlemler Maksimum Minimum Ortalama Std. Sapma lnGSYH 192 11.2938 8.3769 10.1152 0.7009 lnBİT 192 4.3032 3.5204 4.0032 0.1755 lnYET 192 3.9979 1.1568 2.8587 0.8352 Tablo 2, örneklemin 192 gözlemden oluştuğunu, en yüksek standart sapma değerinin lnYE (0.8352) değişkeninde, en düşüğünün ise lnBİT (0.1755) değişkeninde gözlemlendiğini göstermektedir. Tablo 3: Yatay Kesit Bağımlılığı (CDS) Testi Test lnGSYH LnBİT lnYET LM 527.2388 (0.000) 124.3691 (0.000) 364.1493 (0.000) CDLM 66.7135 (0.000) 12.8778 (0.000) 44.9198 (0.000) LMadj 66.5396 (0.000) 12.7039 (0.000) 44.7459 (0.000) CD 22.8432 (0.000) 1.3195 (0.1870) 15.0120 (0.000) Not: Pazantez içindekiler olasılık değerlerini göstermektedir. Tablo 3: Yatay Kesit Bağımlılığı (CDS) Testi Tablo 3 tüm değişkenler için ülkeler arasında yatay kesit bağımlılığı olmadığı yönündeki boş hipotezi reddetmektedir. Başka bir ifadeyle, panelde yatay kesit bağımlılığı vardır. Bu nedenle analize CIPS birim kök testi ile devam edilecektir. Tablo 4: Birim Kök Testleri Değişken Deterministik Düzey 1. Fark Sonuç t-ist. p-değ. t-ist. p-değ. lnGSYH Sabit -2.0361 >=0.10 -2.1509 >=0.10 I (1) Sabit+Trend -2.5911 <0.05 -3.3847 <0.01 lnBİT Sabit -2.0511 >=0.10 -2.3436 >=0.10 I (1) Sabit+Trend -3.7734 <0.01 -4.2489 <0.01 lnYET Sabit -2.4942 <0.05 -2.9543 <0.05 I (0) Sabit+Trend -3.8461 <0.01 -3.8003 <0.01 46 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... Tablo 4’te CIPS birim kök testine göre bağımlı değişken lnGSYH I(1)’de durağan iken, bağımsız değişkenlerden lnBİT, I(1)’de ve lnYET ise I(0)’da durağandır. Buna göre, değişkenlerin hiçbiri CIPS birim kök testini 1’den büyük bir entegrasyon düzeyi ile geçememekte ve bu da panel ARDL yaklaşımının uygulanabilirliğini göstermektedir. Tablo 5: Panel ARDL/PMG Tahmin Sonuçları Uzun Dönem Katsayıları Bağımlı Değişken: lnGSYH Değişken Katsayı Std. Hata t-ist. Olasılık lnBİT 0.8018 0.3462 2.3160 0.0222 lnYET 0.3138 0.1796 1.7471 0.0831 Kısa Dönem Katsayıları Bağımlı Değişken: ΔlnGSYH Değişken Katsayı Std. Hata t-ist. Olasılık -0.2482 0.0749 -3.3135 0.0012 ΔlnGSYH(-1) 0.1937 0.1128 1.7176 0.0883 ΔlnBİT -0.3181 0.1506 -2.1128 0.0366 ΔlnBİT(-1) -0.0815 0.1422 -0.5728 0.5678 ΔlnYET -0.2760 0.1541 -1.7914 0.0756 ΔlnYET(-1) 0.0014 0.0928 0.0155 0.9877 Sabit 1.4801 0.4906 3.0167 0.0031 Trend 0.0090 0.0024 3.7979 0.0002 Not: AIC bilgi kriterine göre optimal gecikme uzunluğu “2” olarak hesaplanmıştır. Δ sembolü fark operatürünü ifade etmektedir. 4. Bulgular ve Tartışma Bu modelde trend spesifikasyonu kapsamında üç farklı modelden yapılan sınamalara göre sabitin ve trendin yer aldığı modelin tahmin edilmesi uygun görülmüştür. Tablo 5: Panel ARDL/PMG Tahmin Sonuçları Not: AIC bilgi kriterine göre optimal gecikme uzunluğu “2” olarak hesaplanmıştır. Δ sembolü fark operatürünü ifade etmektedir. Bu modelde trend spesifikasyonu kapsamında üç farklı modelden yapılan sınamalara göre sabitin ve trendin yer aldığı modelin tahmin edilmesi uygun görülmüştür. Not: AIC bilgi kriterine göre optimal gecikme uzunluğu “2” olarak hesaplanmıştır. Δ sembolü fark operatürünü ifade etmektedir. Bu modelde trend spesifikasyonu kapsamında üç farklı modelden yapılan sınamalara göre sabitin ve trendin yer aldığı modelin tahmin edilmesi uygun görülmüştür. Tablo 5’te, katsayısı, çalışan başına GSYH büyüme oranındaki herhangi bir kısa dönem sapmasının, uzun dönem denge değerine yıllık %24 oranında düzeltildiğini göstermektedir. katsayısının negatif işaretli ve istatistiksel olarak anlamlı olduğu tespit edildiğinden modelin uzun ve kısa dönem katsayıları yorumlanabilmektedir. Buna göre, BİT ve yenilenebilir Tablo 5’te, katsayısı, çalışan başına GSYH büyüme oranındaki herhangi bir kısa dönem sapmasının, uzun dönem denge değerine yıllık %24 oranında düzeltildiğini göstermektedir. katsayısının negatif işaretli ve istatistiksel olarak anlamlı olduğu tespit edildiğinden modelin uzun ve kısa dönem katsayıları yorumlanabilmektedir. Buna göre, BİT ve yenilenebilir Şahin Nas / Ömer Demir \| 47 47 Şahin Nas / Ömer Demir \| 47 Şahin Nas / Ömer Demir \| enerji tüketiminin ekonomik büyümeye etkisi uzun dönemde istatistiksel olarak anlamlı ve pozitiftir. Ancak bu değişkenlerin kısa dönem büyüme etkilerinin anlamlı ve negatif olduğu saptanmaktadır. Uzun dönemde BİT ve yenilenebilir enerji tüketimindeki %1 birimlik artış, çalışan başına GSYH büyüme oranında sırasıyla %0,8 ve %0,3’lük bir artışa neden olmaktadır. BİT ve yenilenebilir enerji tüketiminin pozitif sürdürülebilir büyüme etkisi, Usman vd. (2021) ve Dehmani, Mabrouki & Youssef (2023) ile uyumludur. Büyüme oranını temsil eden çalışan başına GSYH’nin logaritması aynı zaman da teknolojik değişmeyi ifade etmektedir. Bu durumda uzun dönemde BİT ile yenilenebilir enerji tüketimindeki artış ilgili zaman periyodunda BRICS ve MINT ülkelerinde teknolojik değişmeyi tetiklemektedir. Tablo 6: Dumitrescu Hurlin Panel Nedensellik Testi Tablo 6: Dumitrescu Hurlin Panel Nedensellik Testi Null Hypothesis W-İst. Zbar-İst. Olasılık Sonuç lnBİT nedeni değildir lnGSYH 3.6538 1.5136 0.1301 lnGSYİH→lnBİT lnGSYH nedeni değildir lnBİT 4.3533 2.2769 0.0228 lnYET nedeni değildir lnGSYH 7.0615 5.2322 0.0000 lnYET→lnGSYİH lnGSYH nedeni değildir lnYET 3.2811 1.1070 0.2683 lnYET nedeni değildir lnBİT 1.5809 -0.7482 0.4543 - lnBİT nedeni değildir lnYET 2.1591 -0.1172 0.9066 Tablo 6, Dumitrescu-Hurlin panel nedensellik testinin sonuçlarını göstermektedir. 4. Bulgular ve Tartışma Buna göre, lnGSYH ile lnBİT ve lnYET ile lnGSYH arasında tek yönlü bir nedensellik ilişkisi tespit edilmektedir. Ancak lnYET ve lnBİT arasında tek veya iki yönlü bir nedensellik ilişkisi saptanamamıştır. Buradan iki önemli sonuç çıkmaktadır. Birincisi, teknolojik gelişme ortamı BİT artmasına neden olmaktadır. İkincisi ise, yenilenebilir enerji tüketimi çalışan başına GSYH büyüme oranının artmasına neden olmaktadır. 5. Sonuç Ekonomik büyümenin dinamikleri ve faktörleri çok çeşitli ise de ekonomik büyümeyi ölçen kişi başına reel GSYH, ekonomik kalkınmanın en önemli ölçütlerinden biridir. Bu bağlamda bilgi ve iletişim teknolojileri ve yenilenebilir enerji tüketimi gibi unsurlar 1980 sonrası dönemde uluslararası ekonomide önemli büyüme faktörleri haline gelmiştir. Büyüme süreçlerinde BİT, toplam faktör verimliliğini artırarak dünya üretiminin genişlemesinde de önemli bir rol oynamıştır. Bu motivasyon temelinde yapılan bu çalışmada 48 \| Bilgi ve İletişim Teknolojileri, Yenilenebilir Enerji Tüketimi ve Sürdürülebilir Ekonomik... 48 BRICS ve MINT ülkelerinde BİT ve YET faktörlerinin sürdürebilir büyümeyi nasıl ve ne derecede etkilediğini analiz edilmiştir. Bu ülke grubunun seçilmesinin nedeni ise, bu ülkelerin dünya nüfusu ve üretiminin önemli bir kısmını oluşturmasıdır. Bu ülkelerin 1995-2018 dönemine ait BİT ve YET verileri üzerinden PMG tahmincisi kullanılarak Panel ARDL yöntemi uygulanmıştır. Daha sonra ise Dumitrescu-Hurlin panel nedensellik testi yapılmıştır. Analiz sonucunda elde edilen bulgulara göre, 1995-2018 dönemi için BRICS ve MINT ülkelerinde, BİT ve yenilenebilir enerji tüketimi uzun dönemde ekonomik büyümeyi pozitif etkilemektedir. Sonuçlar, BRICS ve MINT ülkelerinde sürdürülebilir büyümeye ulaşmak için BİT’e yapılan yatırımları sürdürmek ve çevre dostu enerji türlerinin tüketimini genişletmek gerektiğini göstermektedir. 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Bu doğrultuda bu çalışma Türkiye ekonomisinde dar para arzı olan M1 para arzı ve işgücü bazlı reel efektif döviz kurunun enflasyon üzerindeki etkisini yıllık frekanslı verilerle 1980-2022 arası dönem için incelemektedir. Bu çalışma kapsamında zaman serisi yöntemleri kullanılmıştır. Serilerin durağanlığı geleneksel ADF testiyle ve yapısal kırılmaları modelleyen Zivot- Andrews birim kök testleriyle sınanmıştır. Seriler arasındaki uzun dönemli ilişkiler ve nedensellik ilişkileri ise güncel ekonometrik yöntemler olan AARDL ve Bootstrap simetrik nedensellik testiyle araştırılmıştır. Düzeyde durağan olmayan serilerin arasında uzun dönemde ‘gerçek’ bir eşbütünleşme ilişkisinin olduğu sonucuna varılmıştır. Dar para arzındaki artışların enflasyon üzerindeki etkisinin istatistiksel olarak anlamlı, negatif fakat nisbi olarak önemsiz olduğu, reel efektif döviz kurundaki artışların ise beklenildiği üzere enflasyonu düşürücü etkiler meydana getirdiği tespit edilmiştir. Bootstrap Simetrik nedensellik analizi sonucuna göre ise, dar para arzı ile enflasyon arasında karşılıklı nedensellik ilişkilerinin olduğu, reel efektif döviz kuru ile enflasyon arasında herhangi bir nedensel ilişkinin olmadığı sonucuna varılmıştır. Sonuç olarak Türkiye ekonomisindeki enflasyonist ortamın döviz kurundaki şoklardan ve yapısal faktörlerden kaynaklandığı çıkarımlarına ulaşılmıştır. https://doi.org/10.58830/ozgur.pub401.c1752 https://doi.org/10.58830/ozgur.pub401.c1752 53 53 54 \| Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 54 1. Giriş En genel tanımıyla fiyatlar genel düzeyindeki sürekli yükselme olarak belirtilen enflasyon, gelişmekte olan bir ülke olarak cari açık, bütçe açığı, işsizlik gibi yapısal sorunlara sahip olan Türkiye Ekonomisinin kronik ve güncel bir problemidir. Son dönemlerde savaş, pandemi ve petrol üretimi kaynaklı enerji krizi, üretim girdi fiyatlarında ve üretim maliyetlerinde artışa neden olmuştur. Pandemi sonrası dönemde uygulanan genişlemeci para ve maliye politikaları ve Rusya-Ukrayna savaşı nedeniyle meydana gelen tahıl ve enerji krizi gibi faktörler Türkiye ekonomisinin kırılgan yapısıyla birleşince enflasyon oranlarının çift hanelere yükselmesine ve kalıcı olmasına neden olmuştur. Enflasyon ekonomik birimlerin alım güçleri üzerinde negatif etkiler oluşturan, hane halklarının yaşam standartlarını düşüren, ekonomide belirsizliği artırarak firma ve yatırımcıların kararlarını güçleştiren bir olgudur. Yüksek ve sürekli bir enflasyon ekonomik birimlerin uzun vadeli kararlar alınmasını engelleyerek, tüketim ve yatırım kararlarını ertelemesine neden olarak, ülke ekonomilerinin sosyal-ekonomik yapısı ve rekabet gücü üzerinde negatif etkiler meydana getirmektedir (Merkez Bankası, 2013:6: Koyuncu, 2014). Fiyat istikrarının olmadığı, yüksek enflasyon oranı, üretim girdi ve çıktıların fiyatlarında yüksek oranda değişikliklere yol açarak üretim sürecinde istikrarsızlığa neden olur. Enflasyonist süreç aynı zamanda spekülatif stoklama sürecini de teşvik ederek bankacılık sisteminin işleyişinde bozukluklara neden olur (Hoai, 2019:83). Enflasyon belirsizliği artırması nedeniyle ekonomik büyümeyi ve mali ve finansal sektörlerin gelişimini, paranın satın alma gücünü azaltması nedeniyle de nüfusun düşük gelirli görece yoksul kesimini olumsuz etkilemektedir. Enflasyonist süreçte paranın satın alma gücü değer kaybeder dolayısıyla insanların harcanabilir gelirleri azalır. Enflasyonist bir ortam bireylerin yaşam kalitesini düşürür, hane haklarının ve toplumun mutluluğunu ve geleceğe yönelik iyimser beklentilerini ortadan kaldırır (Bozkurt, 2014: 310). Enflasyon aynı zamanda ülkelerin ödemeler dengesi üzerinde de etkiler oluşturmaktadır. Fiyatlar genel düzeyinde oluşan yüksek artışlar ilgili ülkenin mal ve hizmetlerine yönelik dış talebi düşürerek ülkenin rekabet gücünü azaltmaktadır. Rekabet gücünün azalması ise ihracatı olumsuz etkileyerek ödemeler dengesinin açık vermesine neden olmaktadır (Merkez Bankası, 2013:6). Bu bağlamda, enflasyon oranı ekonomik birimler üzerindeki negatif etkilerinden dolayı işsizlikle beraber ‘ikiz kötülük’ olarak adlandırılmaktadır (Abel vd., 2021). Bu kapsamda ekonomik birimler üzerindeki negatif etkilerinden dolayı fiyat istikrarını sağlamaya yönelik geliştirilecek politikaların başarısı bir anlamda enflasyonu Sacit Sarı \| 55 55 Sacit Sarı \| Ekonomi yazınında enflasyonu açıklamaya yönelik yaklaşımlar temel olarak üç tür teori ile açıklanmaktadır. Talep teorileri, arz teorileri ve yapısal teoriler (Batarseh, 2021:39). Dash (2017) ise enflasyonu belirleyen teorileri temel olarak iki gruba ayırarak incelemiştir. Olumsuz arz şokları ve yapısal katılık gibi mikroekonomik temelli teoriler ve toplam talep ve genişletici para ve maliye politikasını baz alan makroekonomik faktörler (Dash, 2017: 2). 1. Giriş Paranın Miktar Teorisine dayanan Klasik görüş, ekonomide dışsal olarak belirlenen para arzının enflasyondan birinci derece sorumlu olduğu, para arzında meydana gelen artışların fiyatlar genel düzeyinde aynı oranda artışa neden olacağını öne sürmektedir. Paranın yansız olduğunu belirten bu görüşte, nominal bir değişken olarak para arzında meydana gelen değişmeler reel değişkenleri etkilemeyip yalnızca parasal değişkenleri etkilemektedir. Bu nedenle fiyatlar genel düzeyi para arzı tarafından belirlenmektedir (Cesur, 2006:90). Bu noktada Sabade (2014)’nin vurguladığı gibi Paranın Miktar Teorisi, tam istihdam koşullarında olan bir ekonomide geçerli olabilmektedir. Tam istihdamın olduğu ekonomilerde para arzında meydana gelecek bir artış arzı artırmayıp sadece mal ve hizmetlere yönelik talebi artırmaktadır. Bu durum ise talep çekişli enflasyona neden olmaktadır. Fakat eksik istihdam koşullarının geçerli olduğu az gelişmiş ve gelişmekte olan ülkelerde ise para arzında meydana gelen artışlar üretken alanlara yöneltilerek arzı artırarak ekonominin genişlemesine yardımcı olabilmektedir. Bu da aslında fiyatları yukarı itmek yerine aşağı çekecektir (Sabade, 2014:381). Keynesyen iktisadi görüş ise tam istihdam düzeyinde yer alan bir ekonomide toplam talepte meydana gelecek artışların enflasyona neden olacağını belirtmektedir (Duodu vd., 2022: 2-3). Kısa dönemde ücretlerin katı olduğu varsayımıyla, tam istihdam düzeyinde faaliyette olan bir ekonomide talepte meydana gelen bir artış sabit ücret varsayımıyla firmaların kârlarını artırmalarına neden olacaktır. Artan talebi karşılamak için işgücü talebi artan firmalar piyasada ücretlerin, dolayısıyla maliyetlerin yükselmesine neden olacaktır. Bu durum ise fiyatlar genel düzeyini artırarak enflasyona yol açacaktır (Koyuncu, 2014:226). Enflasyonun her yerde ve her zaman ‘parasal’ bir olgu olduğunu belirten Monetarist yaklaşım ise açıklamalarını Modern Miktar Teorisi bağlamında yapmaktadır. Enflasyonun yalnızca para arzındaki artışların ekonominin çıktı seviyesinden fazla olduğunda, para arzı büyümesinin ekonomik büyümeyi aşması halinde enflasyona neden olacağını öne sürmüşlerdir (Su vd., 2016:702: Dash,2017:2). Yapısalcı yaklaşım ise özellikle az gelişmiş ve gelişmekte olan ülkelerde enflasyonun ilgili ülkenin ekonomik ve sosyal yapısından kaynaklandığını öne sürmektedir. İlgili ülkelerdeki düşük verimlilik, yolsuzluk, ödeme açıkları, ürün tedarik zincirindeki aksaklıklar ve siyasal ve politik krizler enflasyondan sorumlu Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 56 olmaktadır. Gelişmekte olan ülkelerin bazı sektörlerinde piyasa eksikliklerinin ve yapısal dengesizliklerin varlığının arz ve talep arasında uyumsuzluk yarattığını ve bu durumun enflasyonu artırdığı belirtilmiştir (Sultana vd., 2019:43: Smauel vd., 2019). Para arzındaki artışların özellikle tam istihdam koşullarına sahip gelişmiş ülkelerde paranın miktar teorisi uyarınca enflasyonu artırdığı literatürde üzerine görüş birliği varılan bir konudur. Türkiye ekonomisi gibi âtıl kapasitenin olduğu, gelişmekte olan ülkelerde ise para arzındaki artışlar toplam talepte canlanmaya neden olarak fiyatlar genel düzeyini artırmayabilmektedir. 1. Giriş Bu çalışma kapsamında, gelişmekte olan bir ülke olarak Türkiye ekonomisinde para arzındaki artışların enflasyona neden olup olmadığı 1980-2022 arası dönem için incelenecektir. Bu çalışma güncel bir ekonometrik yöntem olan AARDL yöntemini enflasyon ve para arzı bağlamında inceleyen ilk çalışma olması nedeniyle literatüre katkı sunması beklenmektedir. Çalışma giriş bölümünden sonra Türkiye ekonomisinde 1980-2022 arası enflasyonun izlediği seyri tartışacaktır. Literatür taraması ve Veri Seti ve Metodoloji bölümlerinde sırasıyla, enflasyon üzerine seçilmiş çalışmaların sonuçları özetlenerek, kullanılan veri seti ve yöntemler tanıtılacaktır. Bulgular bölümünde ise analiz sonuçları sunularak çıkarımları tartışılacaktır. Çalışma Sonuç ve Politika Önerileri bölümüyle tamamlanacaktır. 1.1 Türkiye Ekonomisinde Enflasyonun Seyri Türkiye ekonomisinde enflasyon oranının 1980-2022 arası dönemde izlediği seyir Şekil- 1 aracılığıyla verilmiştir. 24 Ocak 1980 kararlarıyla ithal ikameci politikaların terk edilerek, serbest dış ticaret politikalarına yönelik politikalar geliştirilen Türkiye ekonomisinde ilgili dönemde sabit kur rejimi terk edilerek sürünen pariteye geçilmiştir. Sermaye hareketleri ve finansal piyasaların liberalizasyonu yönünde adımlar atılmıştır. (Merkez Bankası, 2013). Pamuk (2022) ifadesiyle ilgili dönemdeki temel sorun kamu maliyesinden dengeleri sağlamadan sermaye hareketlerinin serbest bırakılması ekonomiyi dış şoklara karşı çok daha açık ve kırılgan yapmıştır (Pamuk, 2022:283). Nitekim enflasyon oranı izleyen dönemlerde çift hanelerde gerçekleşmiştir. Enflasyonun yüksek oranlardan seyrettiği 1990’lı yıllar ise ekonomik ve politik krizlerin yaşandığı, borç stokunun yükseldiği, ekonomik ve siyasi istikrarsızlıklar nedeniyle ekonomideki kırılganlıkların arttığı bir dönem olmuştur. 1994 yılı Türkiye ekonomisinde en yüksek enflasyonun gerçekleştiği dönem olmuştur (Merkez Bankası, 2013: 15). 2001 krizi sonrası Türkiye ekonomisinde birçok alanda yapısal reformlar yapılmıştır. ‘Güçlü Ekonomiye Geçiş Programı’ ile makroekonomik istikrarın sağlanması amaçlanarak fiyat istikrarını sağlamanın Merkez Bankasının Sacit Sarı \| 57 Sacit Sarı \| 57 Sacit Sarı \| 57 57 temel amacı olduğu vurgulanmıştır (Kılavuz ve Altınöz, 2020: 243). Başarılı bir şekilde uygulanan para ve maliye politikaları neticesinde enflasyon oranlarında önemli düzeyde azalışlar meydana gelerek uzun bir aranın ardından 2004 yılında tek haneli enflasyon oranı gerçekleşmiştir. Bozkurt (2014)’un vurguladığı gibi 2002 yılında uygulanmaya başlanan örtük enflasyon hedeflemesi ve buna tamamlayıcı olarak 2006 yılında uygulanmaya başlanan açık enflasyon hedeflemesi enflasyonu düşük seviyelerde tutma konusunda istikrarı sağlamada başarılı olmuştur. 2004-2017 arası oldukça istikrarlı seyreden enflasyon oranı daha sonraki süreçte yerli parada meydana gelen önemli düzeydeki değer kayıpları ve petrol krizi, pandemi ve savaş koşulları nedeniyle tekrardan artış trendine girmiştir. 2022 yılında enflasyon oranı %72.30 oranında gerçekleşmiştir. Şekil 1: Türkiye Ekonomisinde Enflasyonun Seyri Şekil 1: Türkiye Ekonomisinde Enflasyonun Seyri 2. Literatür Taraması Literatürde enflasyon üzerine çok sayıda çalışma yapılmasına rağmen, enflasyonun ekonominin geneli, ekonomik birimler üzerindeki negatif etkilerinin büyüklüğü ve geliştirilen yeni ekonometrik yöntemler sayesinde enflasyonu farklı bakış açılardan ve yöntemlerden inceleyen çalışmalara ihtiyaç duyulmaktadır. Bu bölüm kapsamında öncelikle yabancı ülke (ler) üzerine yapılan çalışmaların sonuçları Tablo-1’de özetlenecektir. 58 \| Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 58 Tablo 1: Enflasyon Para Arzı İlişkisini inceleyen Çalışmalar (Dış Ülke(ler)) Çalışma Künyesi Dönem Aralığı/ Ülke Kullanılan Yöntem Sonuç McCandless ve Weber / 1995 1960-1990 / 110 Ülke Ekonomisi Korelasyon Analizi Para arzı ile enflasyon arasında pozitif ve yüksek oranda korelasyon mevcuttur. Uddin vd. /2014 1972-2012/ Bangladeş Ekonomisi ARDL sınır testi Enflasyon ve seçili değişkenler eşbütünleşiktir. Para arzı enflasyon üzerinde pozitif etkilere sahiptir. Jiang, Chang ve Li / 2015 1999:01- 2014:06 / Çin Ekonomisi Dalgacık (Wavelet) Analizi Kısa dönemde M1 para arzı enflasyon üzerinde negatif etkiler oluşturmaktadır. Nguyen /2015 1985-2012 / 9 Asya Ülke Ekonomisi Panel GMM Para arzı enflasyon üzerinde pozitif anlamlı etkilere sahiptir. Diermeier ve Goecke / 2016 1999:Q1- 2016Q1 / Euro Bölgesi Korelasyon Analizi ve Granger Nedensellik Analizi Para arzı ile enflasyon arasında Granger nedensellik ilişkisi mevcuttur. Bekiros vd. /2017 1999:11- 2014:11 / Hindistan, Malezya ve Japonya Ekonomisi Zamanla Değişen Nedensellik Analizi Enflasyondan para arzına doğru nedensellik ilişkisi olduğu sonucuna varılmıştır. Dash /2017 1971-2010/ Hindistan Ekonomisi VAR ve Hata Düzeltme Modeli Para arzı (özellikle M1) enflasyon üzerinde anlamlı pozitif etkilere sahiptir. Kısmi Paranın Miktar Teorisi geçerlidir. Amassoma vd. / 2018 1970-2016 / Nijerya Ekonomisi Eşbütünleşme ve Granger Nedensellik Analizi Para arzının enflasyon üzerinde bir etkisi bulunmamaktadır. Hoai / 2019 1995-2016 / Vietnam Ekonomisi Yapısal Eşitlik Modeli Döviz kuru, faiz oranı ve para arzı enflasyon üzerinde pozitif etkilere sahiptir. Smauel vd. / 2019 2010:01- 2018:12 / Nijerya Ekonomisi Eşbütünleşme ve Granger Nedensellik Testi Enflasyon parasal faktörlerden çok yapısal faktörlerden kaynaklanmaktadır. Ahmad ve Yahaya / 2021 1980-2019 / Birleşik Krallık Ekonomisi ARDL sınır testi Para arzındaki artışlar enflasyonu artırmaktadır. Batarseh / 2021 1980-2019 / Ürdün Ekonomisi Eşbütünleşme ve Granger Nedensellik Analizi M1 para arzı ile enflasyon arasında eşbütünleşme ilişkisi yoktur. M1 para arzından enflasyona doğru tek yönlü Granger nedensellik ilişkisi mevcuttur. Tablo 1: Enflasyon Para Arzı İlişkisini inceleyen Çalışmalar (Dış Ülke(ler)) Sacit Sarı \| 59 Sacit Sarı \| 59 59 Tablo-2 ise çalışmanın konusunu oluşturan Türkiye ekonomisinde enflasyon üzerine yapılan çalışmaların sonuçları özetlenmiştir. 2. Literatür Taraması Çalışmalarda genel olarak para arzı ve enflasyon arasında uzun dönemli ilişki olduğu ve para arzındaki artışların enflasyonu artırdığına dair bulgular tespit edilmiştir. Tablo 2: Enflasyon Para Arzı İlişkisini inceleyen Çalışmalar (Türkiye Ekonomisi) Çalışma Künyesi Dönem Aralığı/ Ülke Kullanılan Yöntem Sonuç Cesur / 2006 1994:1-2004:1 / Türkiye Ekonomisi Regresyon Analizi Para arzındaki artışlar enflasyonu artırmaktadır. Çiçek / 2011 1987Q1- 2007Q3 / Türkiye Ekonomisi Eşbütünleşme ve Granger Nedensellik Para arzı ve enflasyon arasında çift yönlü Granger nedensellik ilişkisi tespit edilmiştir. Bozkurt / 2014 1999:Q2- 2012:Q4/ Türkiye Ekonomisi Eşbütünleşme ve VAR analizi Para arzı ve paranın değişim hızı enflasyonun temel belirleyicileridir. Koyuncu / 2014 1987:01- 2013:04 / Türkiye Ekonomisi Eşbütünleşme ve Granger Nedensellik Bütçe açığı, M2 para arzı ve enflasyon serileri eşbütünleşik değildir. Para arzından Enflasyona doğru tek yönlü Granger nedensellik ilişkisi mevcuttur. Gabaçlı /2020 2000Q1- 2017Q2 / Türkiye Ekonomisi Eşbütünleşme ve Granger Nedensellik M2 para arzından enflasyona doğru tek yönlü Granger nedensellik ilişkisi vardır. Kılavuz ve Altınöz / 2020 2006:Q4- 2018:Q4 / Türkiye Ekonomisi ARDL sınır testi M1 para arzının enflasyon üzerinde negatif fakat anlamsız bir etkisi bulunmaktadır. Bozkurt / 2021 2011:01- 2021:08 / Türkiye Ekonomisi VAR analizi ve Granger Nedensellik Testi Faiz oranları döviz kurunun, döviz kuruda enflasyonun Granger nedenidir. Tablo 2: Enflasyon Para Arzı İlişkisini inceleyen Çalışmalar (Türkiye Ekonomisi) (1) (1) 3. VERİ SETİ VE METODOLOJİ Türkiye Ekonomisinde dar para arzı (M1) ile enflasyon arasındaki ilişkiyi 1980-2022 arası dönemde yıllık frekanslı verilerle inceleyen bu çalışmada kullanılan değişkenlere ait tanımlayıcı istatistikler Tablo-3 aracılığıyla Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 60 verilmiştir. M1 para arzı dışındaki diğer seriler normal dağılmaktadırlar. Dolaşımdaki kâğıt paralarla madeni paraların toplamından banka kasalarındaki nakitin çıkarılmasıyla elde edilen M0 para arzına vadesiz mevduatların eklenmesiyle oluşan M1 para arzı, Cukierman (2017) vurguladığı gibi enflasyondaki değişimi en iyi yansıtan para arzıdır. Bu doğrultuda para arzını temsilen M1 para arzı seçilmiştir. Tablo 3: Tanımlayıcı İstatistikler İstatistik Enflasyon (LENF) M1 Para Arzı (LM1) Reel Efektif Döviz Kuru(LREX) Ortalama 39.39 2.40E+11 86.89 Minumum 6.25 1086939 41.54 Maksimum 105.21 3.12E+12 115.08 Standart Hata 30.24 5.90E+11 17.10 J-B Normallik Olasılık Değeri 0.15 0.00 0.32 Veri Kaynağı Dünya Bankası FRED Economic Data FRED Economic Data Tablo 3: Tanımlayıcı İstatistikler Tablo 3: Tanımlayıcı İstatistikler Türkiye ekonomisi gibi enerjide dışa bağımlı ve ihracat ürünlerinde yüksek oranda ithal ara malı kullanan ülkelerde döviz kurunun da enflasyon üzerinde etkiler oluşturması beklenmektedir. Teorik olarak yerli parada meydana gelen değer kayıplarının ithal mallarını nispi olarak pahalılaştıracağı için enflasyonu artırması beklenmektedir. Modele ilave edilen işgücü bazlı reel efektif döviz kuru değişkeni ise yerli parada meydana gelen artışlar sonucu değer kazanmaktadır. Bu bağlamda değer kazanan bir yerli para yüksek reel efektif döviz kuru anlamına gelmektedir. İlgili katsayının negatif olması beklenmektedir. Nitekim yüksek bir reel efektif döviz kuru yerli paranın değer kazandığını ve ithal malların nisbi olarak ucuzladığı anlamına gelmektedir. Serilere ait logaritmik dönüşüm gerçekleştirilmiştir. Çalışma kapsamında para arzının ve reel efekti döviz kurunun enflasyon üzerindeki etkisini incelemek için Model-1 tahmin edilecektir. Çalışmada zaman serisi yöntemleri kullanılmaktadır. Serilerin durağanlığı geleneksel ADF birim kök testi (BKT) ve yapısal kırılmalı Zivot-Andrews (1992) BKT ile sınanacaktır. Seriler arasındaki uzun dönemli ilişkiler ise Peseran Shin ve Smith (2001) tarafından geliştirilen ARDL sınır testini genişleten Sam, McNown ve Goh (2019)’un literatüre kazandırdığı Augmented (Geliştirilmiş) ARDL (AARDL) sınır testi ile araştırılacaktır. \| 61 61 Sacit Sarı Bu yaklaşımın diğer eşbütünleşme testlerine göre birtakım avantajları bulunmaktadır. ARDL sınır testinde tüm değişkenlerin içsel olduğu varsayılmaktadır. Model kapsamında kısa ve uzun dönem katsayılar eşanlı olarak tahmin edilebilmektedir. Son olarak ARDL sınır testi bu çalışmada olduğu gibi küçük örneklem boyutlarında daha tutarlı ve etkili sonuçlar vermektedir (Dritsakis, 2011:4-6). Son olarak seriler arasındaki dinamik nedensellik ilişkileri Hacker ve Hatemi-J (2006) tarafından geliştirilen Bootstrap Simetrik Nedensellik testiyle incelenecektir. AARDL sınır testi diğer eşbütünleşme testleriyle karşılaştırıldığında durağanlık özellikleri bakımından daha esnektir. Değişkenler I(0) ve/veya I(1) olabilmektedir. Eşbütünleşmenin varlığı F OVERALL, t DV ve F IDV sınır testleri ile tespit edilmiştir (Balkı ve Göksu, 2023: 203). Uzun dönemli ilişkisinin varlığı Eşitlik-2’de verilen denklem aracılığıyla sınanacaktır. (2 (2) F OVERALL = = ; = t DV = = =0 ; = 0 F IDV = ; Her üç test istatistiği kapsamında da test istatistikleri kritik değerlerden büyük olduğunda temel hipotezler reddedilerek değişkenler arasında ‘gerçek’ bir eşbütünleşme ilişkisinin varlığından bahsedilebilmektedir. 4. BULGULAR Değişkenlere ait ADF BKT sonuçlarına Tablo-4’te yer verilmiştir. Yapısal kırılmaların modellenmediği bu test kapsamında tüm serilerin birim kök içerdiği, LENF değişkenin farkı alınınca durağanlaştığı, diğer değişkenlerin ise yalnızca sabit terimli modelde %10 anlam düzeyinde durağanlaştığı tespit edilmiştir. 62 \| Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 62 Tablo 4: ADF BKT Sonucu Değişken Sabit Terimli Model Sabit ve Trendli Model LENF -1.4602 -0.4334 LM1 -1.3617 -1.3647 LREX 0.1593 0.3618 LENF -4.3777* -4.4411* LM1 -2.6884* -2.9069 LREX -2.8761* 0.0681 Not: , *, %10 ve %1 anlam düzeylerini ifade etmektedir. Tablo 4: ADF BKT Sonucu Tablo 4: ADF BKT Sonucu Not: , *, %10 ve %1 anlam düzeylerini ifade etmektedir. Türkiye ekonomisinde çalışma dönemi olan 1980-2022 aralığı politik, ekonomik ve küresel krizler nedeniyle yapısal kırılmaların meydana geldiği bir dönem olarak dikkat çekmektedir. Bu bağlamda serilerin durağanlıkları ayrıca Zivot-Andrews (1992) BKT ile sınanmıştır. Bu teste ait sonuçlar ise Tablo-5 aracılığıyla verilmiştir. Tablo 5:Zivot-Andrews BKT Sonucu Değişken Test İstatistiği Kırılma Tarihi LENF -2.5719 2003 LM1 -4.2946 1994 LREX -1.3118 2016 Not: Model A % 5 Tablo Kritik değeri=-4.93 Tablo 5:Zivot-Andrews BKT Sonucu Not: Model A % 5 Tablo Kritik değeri=-4.93 Zivot-Andrews BKT sonuçlarına göre değişkenlere ait test istatistiğinin kritik değerden (-4.93) büyük olması nedeniyle durağan olmadığı (birim kök içerdiği) sonucuna varılmıştır. Kırılma tarihleri olarak ise Türkiye ekonomisinde ekonomik krizin (1994), Politik krizin (2016 Askeri Darbe Girişimi), ve yapısal dönüşümlerin başladığı dönem olan 2003 tarihleri tespit edilmiştir. Birim kök içeren seriler arasındaki uzun dönemli dinamik ilişkiler ise AARDL yöntemiyle sınanmıştır. AARDL testine ait tüm sonuçlar Tablo-6’da verilmiştir. Öncelikle eşbütünleşme ilişkisinin varlığının sınandığı F OVERALL, t DV ve F IDV test sonuçlarına göre test istatistiklerinin sınır değerlerinin üzerinde olmasından dolayı seriler arasında eşbütünleşme ilişkisinin var 63 Sacit Sarı Sacit Sarı olduğu sonucuna ulaşılmıştır. Bu bağlamda Türkiye ekonomisinde 1980- 2022 arasın dönemde enflasyon oranı, M1 para arzı ve Reel efektif döviz kuru arasında uzun dönemli ‘gerçek’ bir ilişki mevcuttur. Uzun dönem katsayı sonuçlarına göre ise LM1 para arzında meydana gelen artışlar enflasyon üzerinde nisbi olarak küçük fakat istatistiksel olarak anlamlı ve negatif etkiler oluşturmaktadır. LM1 para arzında meydana gelen %1’lik artışlar enflasyonu %0.08 gibi düşük bir oranda azaltmaktadır. Tablo 6: AARDL Sonucu Tablo 6: AARDL Sonucu Eşbütünleşme Test İstatistikleri Eşbütünleşme Testi Test İstatistiği I(0) %5 I(1) %5 Foverall 16.8445 3.548 4.803 tDV -4.9136 -2.86 -3.78 FIDV 14.2125 2.85 4.97 Uzun Dönem Katsayılar Değişken Katsayı Standart H. 4. BULGULAR t istatistiği Olasılık Değeri LM1 -0.0875 0.0115 -7.5949 0.0000 LREX -1.1010 0.5343 -2.0605 0.0478 Kısa Dönem Katsayılar ve Hata Düzeltme Terimi Değişken Katsayı Standart H. t istatistiği Olasılık Değeri C 6.3503 0.8725 7.2777 0.0000 D(LENF(-1)) 0.3305 0.1242 2.6611 0.0122 D(LENF(-2)) 0.2996 0.1328 2.2550 0.0313 D(LM1) -0.3399 0.3260 -1.0425 0.3052 D(LM1(-1)) 1.3081 0.3348 3.9065 0.0005 ECM(-1) -0.7166 0.0977 -7.3344 0.0000 Tanısal Testler Tanısal Test F istatistiği Olasılık Değer Ramsey RESET 0.8252 0.3709 Breusch-Godfrey Otokorelasyon 0.6330 0.5381 Breusch-Pagan- Godfrey Değişen Varyans 1.1657 0.3503 Jarque-Bera Normallik Testi 1.6645 0.4350 Not: Foverall test istatistiği Narayan (2005), tdv istatistiği, Peseran vd. (2001), FIDV test istatistiği ise Sam vd. (2019)’dan alınmıştır. Not: Foverall test istatistiği Narayan (2005), tdv istatistiği, Peseran vd. (2001), FIDV test istatistiği ise Sam vd. (2019)’dan alınmıştır. Not: Foverall test istatistiği Narayan (2005), tdv istatistiği, Peseran vd. (2001), FIDV test istatistiği ise Sam vd. (2019)’dan alınmıştır. Teoride beklenen etkinin aksi olarak çıkan bu sonuç, Sabade (2014) tarafından belirtildiği üzere eksik istihdamda çalışan ekonomilerde, para arzının enflasyon üzerinde negatif etkiler meydana getirmesi, beklenen olası bir sonuç olabilmektedir. Bu tür ülkelerde para arzında meydana gelen Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 64 artışlar, ekonominin âtıl alanlarına yönelerek ekonominin üretim kapasitesini genişlemesine yardımcı olarak talep baskı meydana getirmeyerek enflasyona yol açmayabilmektedir. Fakat ilgili katsayı görece olarak düşük düzeyde gerçekleşmiştir. Bu sonuç literatürde Jiang vd. (2015) ve Kılavuz ve Altınöz (2020) sonuçlarıyla paralellik göstermektedir. Bir diğer açıklayıcı değişken olan LREX değişkeni ise enflasyon üzerinde negatif etkiler meydana getirmektedir. Ülke parasında meydana gelen değerlenmeler sonucu yükselen Reel Efektif Döviz Kuru, ithal malların nisbi fiyatlarını ucuzlatarak enflasyon üzerinde negatif etkiler oluşturmaktadır. Aksine döviz kurunda yaşanan değerlenmeler sonucu (yerli paranın değer kaybetmesi) LREX değişkeninin değer kaybetmesi anlamına geleceğinden, ithal malların görece fiyatını pahalılaştırarak enflasyon üzerinden pozitif etkiler oluşturacaktır. Bu kapsamda ilgili sonucun teorik olarak beklenen bir sonuç olduğunu söylenebilmektedir. Bu sonuç ayrıca Hoai (2019) ve Bozkurt (2021) çalışmalarıyla benzer sonuca varmıştır. Kısa dönemde ise enflasyon kendi gecikmeli değerinden pozitif yönde etkilenmektedir. Bu doğrultuda enflasyonun kendi kendini besleyen bir süreç olduğu sonucuna varılmıştır. Kısa dönemde para arzındaki artışların ise enflasyon üzerindeki etkisinin istatistiksel olarak anlamsız olduğu tespit edilmiştir. ARDL sınır testinde uzun dönem dengesinden meydana gelen sapmaların dengeye gelip gelmedikleri ECM (Hata Düzeltme Terimi) ile incelenmektedir. İlgili katsayının istatistiksel olarak anlamlı ve 0 ile -1 arasında değer alması beklenmektedir. 4. BULGULAR Bu bağlamda ECM katsayısının istatistiksel olarak anlamlı olduğu ve kısa dönemde dengeden meydana gelen sapmaların yaklaşık olarak %71’i bir dönem sonra dengeye gelmektedir. AARDL sonuçlarının tutarlılığı ve etkinliği için modelin bir takım tanısal testlerden geçmesi gerekmektedir. Bu doğrultuda modelde spesifikasyon hatası olmadığı, serilerin normal dağıldığı, otokorelasyon içermediği ve değişen varyans sorununun olmadığı yapılan testlerle ortaya konulmuştur. Son olarak katsayıların istikrarlığı ve modelin öngörü amacıyla kullanılıp kullanılmayacağı Cusum ve Cusum Square testleriyle incelenmiştir. Şekil- 2’de verildiği üzere katsayıların güven aralığı içinde olduğu, modelin istikrarlı, tutarlı olduğu, yapısal kırılmanın gerçekleşmediği ve modelin öngörü amacıyla kullanılabileceği sonucuna varılmıştır. 65 Sacit Sarı -20 -15 -10 -5 0 5 10 15 20 92 94 96 98 00 02 04 06 08 10 12 14 16 18 20 22 CUSUM 5% Significance -0.4 -0.2 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 92 94 96 98 00 02 04 06 08 10 12 14 16 18 20 22 CUSUM of Squares 5% Significance Şekil:2 Cusum ve Cusum Square Sonucu Şekil:2 Cusum ve Cusum Square Sonucu Seriler arasındaki dinamik nedensellik ilişkisi ise Hacker ve Hatemi-J (2006) tarafından geliştirilen Simetrik Nedensellik testiyle sınanmıştır. Değişkenlerin düzey değerleriyle analize dahil edildiği bu test Toda- Yamamoto (1995) test mantığına benzemektedir. Değişkenlerin düzey değerleriyle analize tabi tutulduğu bu testte teste ait kritik değerler Bootstrap yöntemiyle elde edilmektedir. Tablo-7’de verilen sonuçlara göre para arzı ile enflasyon arasında karşılıklı nedensellik ilişkilerin olduğu, Reel efektif döviz kuru ile enflasyon arasında ise herhangi bir nedensellik ilişkisinin olmadığı sonucuna varılmıştır. LM1 ile LENF arasında tespit edilen karşılıklı nedensellik Çiçek (2011) sonucuyla benzer, para arzından enflasyona doğru tek yönlü nedensellik olduğunu belirten Batarseh (2021) çalışmasının aksi yönde çıkmıştır. Tablo 7: Hacker ve Hatemi-J Simetrik Nedensellik Test Sonucu Nedenselliğin Yönü Wstat Bootstrap Değeri %5 Sonuç LM1 LENF 37.029 9.137 Nedensellik var. LENF LM1 19.614 8.151 Nedensellik var. LREX LENF 3.668 4.280 Nedensellik yok. LENF LREX 0.0000 4.100 Nedensellik yok. Tablo 7: Hacker ve Hatemi-J Simetrik Nedensellik Test Sonucu 5. Sonuç ve Politika Önerileri Ekonomide belirsizliği artırarak, hane halklarının alım gücünün düşmesine, gelir dağılımının kötüleşmesine, firmaların yatırım kararlarını ertelemesine veya iptal etmesine neden olan enflasyon oranı işsizlikle Türkiye Ekonomisinde Dar Para Arzı- Enflasyon İlişkisinin AARDL Yöntemiyle Sınanması 66 beraber bir ekonomide en çok istenmeyen makro ekonomik sorunlardan biridir. Ekonominin hem makro hem de mikro kesimi üzerinde oluşturduğu negatif etkilerden dolayı enflasyon oranı, kamuoyu, araştırmacılar ve politika yapıcılar tarafından yakından izlenen bir olgudur. Bu bağlamda fiyat istikrarını sağlamaya yönelik olarak geliştirilecek para ve maliye politikalarının başarısı bir anlamda enflasyonu ve belirleyen faktörlerin doğru şekilde tespit edilmesiyle mümkün olacaktır. Enflasyonu belirleyen faktörler ise genellikle talep çekişli, maliyet itişli ve yapısal faktörlerden kaynaklanmaktadır. Bu bağlamda bu çalışma Türkiye ekonomisinde dar para arzı olan M1 para arzı ve işgücü bazlı reel efektif döviz kurunun enflasyon üzerindeki etkisini yıllık frekanslı verilerle 1980-2022 arası dönem için güncel bir ekonometrik yöntem olan AARDL ve Bootstrap Nedensellik testiyle araştırmıştır. Düzeyde durağan olmayan seriler arasında ‘gerçek’ bir eşbütünleşme ilişkisinin varlığının tespit edildiği çalışmada, kısa dönemde enflasyonun kendi gecikmeli değerlerinden pozitif yönde etkilendiği sonuçlarına varılmıştır. Uzun dönemde ise teoride beklenildiği üzere döviz kurunda yaşanan değerlenmeler (yerli paranın değer kaybetmesi / reel efektif döviz kurunun düşmesi) enflasyon üzerinde pozitif etkiler yapmaktadır. Literatürde yapılan çalışmaların aksine ise dar para arzında meydana gelen artışlar enflasyon üzerinde nisbi olarak küçük fakat negatif yönde etkiler oluşturmaktadır. Bootstrap Simetrik Nedensellik sonucu ise dar para arzı ile enflasyon arasında karşılıklı nedensellik ilişkisinin olduğu, reel efektif döviz kuru ile enflasyon arasında ise herhangi bir nedensellik ilişkinin olmadığı sonucuna varılmıştır. Bu doğrultuda Paranın Miktar Teorisinin önermesinin Türkiye ekonomisi için geçerli olmadığı, ekonominin eksik istihdamda bulunduğu için para arzında meydana gelen artışların Keynesyen iktisadi ekol uyarınca ekonomide talebi canlandırıcı etkiler oluşturduğu sonucuna varılmıştır. Döviz kurundaki dalgalanmaların enflasyon üzerinde etkiler oluşturduğu Türkiye ekonomisinde, kur şoku nedeniyle yerli parada yaşanacak değer kayıplarının ithal malların fiyatını pahalılaştırarak beklenildiği üzere enflasyonu artırdığı görülmektedir. 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Determinants of inflation in Bangladesh: An econometric investigation. Journal of World Economic Research, 3(6), 83-94. Bölüm 5 Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme Bengü Kayapalı Aslan1 Özet Gelir, bir ekonomide bir dönem içerisinde biriken para, mal veya hizmetlerin bütünüdür. Hızla gelişen ülke ekonomileri göz önünde bulundurulduğunda ortaya çıkan bu bütünün nasıl bölüşüleceği önemli bir sorundur. Bu noktada ortaya çıkan gelir dağılımı kavramı çalışmanın ana konusunu oluşturmaktadır. Çünkü gelir dağılımı öncelikle ülkelerin ekonomik büyümeleri sürecinde elde ettikleri varlıklarını dengeli biçimde dağıtmaları konusunda önemli rol üstlenmektedir. Bu noktada adaletin sağlanması ülkenin gelişmiş düzeyinin belirlenmesinde etkilidir. Gelir dağılımına ilişkin göstergelerin, belirleyicilerin, türlerin ve iktisadi ekollerin yaklaşımlarının incelendiği bu çalışmada öncelikli olarak gelir dağılımı kavramı açıklanmaktadır. Daha sonra gelir dağılımına ilişkin oluşan literatür incelenerek, gelir dağılımı konusunda yeterli düzeyde çalışmalar yapılıp yapılmadığı noktasında tespitte bulunulacaktır. Çalışmanın sonuç bölümünde ise gelir dağılımı konusunu inceleyen çalışmalara derinlik kazandırmak ve ülkelerdeki gelir dağılımı eşitsizliği sorununun çözümüne ilişkin politika öneri sunulacaktır. Giriş Gelirin belirli kişi ve grupların elinde toplanması, toplum içerisindeki adalet duygusunu zedelemektedir. Toplu yaşam içerisinde insanların günlük ihtiyaçlarını karşılamalarındaki yegâne kaynak olan gelirin dağılımı, birçok disiplinin araştırma konusu olmuştur. Para, mal veya hizmet akışı üzerinden tanımlanan gelir, belirli bir dönemde bir ekonomi içerisindeki bir kişi, grup veya şirket tarafından üretilmektedir. Bir ekonomi içerisinde belli bir dönemde elde edilen gelirin, geliri elde edenler arasında nasıl bölüşüleceği https://doi.org/10.58830/ozgur.pub401.c1753 https://doi.org/10.58830/ozgur.pub401.c1753 71 72 \| Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 72 ise gelir dağılımı kavramının kapsamı içerisindedir. Bir ülkedeki gelirin çeşitli gruplar arasındaki bölüşümü olarak tanımlanan gelir dağılımı, belirli faktörlerce etkilenmektedir. Gelir dağılımına ilişkin çalışmalarda her ne kadar demokratikleşme ve küreselleşme olguları öne çıksa da gelirin dağılımını etkileyen birçok faktör bulunmaktadır. Bu faktörler; ekonomik büyüme, küreselleşme, teknoloji, üretim faktörlerinin dağılımı vb. olarak sıralanmaktadır. Gelir dağılımını etkileyen faktörler aynı zamanda gelir dağılımı türlerinin oluşmasında da pay sahibidir. Kişisel gelir dağılımı, fonksiyonel gelir dağılımı, bölgesel gelir dağılımı, sektörel gelir dağılımı vb. türleri olan gelir dağılımının belirli ölçüm yöntemleri bulunmaktadır. Gelir dağılımında öne çıkan bir kriter olarak bilinen Gini Katsayısının yanında, Pareto Katsayısı, Atkinson Eşitsizlik Ölçütü ve Theil Endeksi gibi birçok ölçüt gelir eşitsizliğinin belirlenmesinde kullanılmaktadır. Gelir dağılımı iktisat bilimi içerisinde yer alan ekollerce de farklı perspektiflerde ele alınmıştır. Bu noktada; Merkantalistlerin ve Fizyokratların Gelir Dağılımı Yaklaşımı, Klasik İktisadın Gelir Dağılımı Yaklaşımı, Neo-Klasik İktisadın Gelir Dağılımı Yaklaşımı, Keynesyen Gelir Dağılımı Yaklaşımı, Marksist İktisadın Gelir Dağılımı Yaklaşımı birbirlerinden farklılıklar içerdiği için hepsinin ayrı ayrı irdelenmesi gerekmektedir. Gelir dağılımına ilişkin makale, tez, rapor, ihtisas komisyonu çalışmaları, uluslararası örgütlerin deklare ettikleri raporlar ve araştırma bulguları başta olmak üzere birçok akademik ve kurumsal çalışma yer almaktadır. Bu da gelir dağılımına ilişkin zengin bir literatürün olduğunu göstermektedir. Bu çalışmanın amacı, öncelikle gelir dağılımı kavramını açıklamaktır. Bu açıklama gelir dağılımı türleri, gelir dağılımını etkileyen faktörler, gelir dağılımı eşitsizliği ölçüm yöntemleri ve iktisadi ekollerin gelir dağılımına yaklaşımları başlıklarıyla yapılmaktadır. Çalışmada daha sonra gelir dağılımına ilişkin literatür incelenerek (makale, tez, vb.), gelir dağılımı konusunun yeterli düzeyde incelenip incelenmediği sorgulanacaktır. Çalışmanın sonuç bölümünde ise öncelikle gelir dağılımı kavramı konu alan çalışmalarına derinlik kazandırılması amacıyla birtakım öneriler sunulacaktır. Bunun yanında ülkelerin gelir dağılımı adaletsizliğine yönelik politika geliştirmeleri noktasında çözüm önerileri sunulacaktır. Gelir ve Gelir Dağılımı Kavramı Gelir dağılımı kavramının daha iyi anlaşılması için öncelikle gelir kavramının açıklanması gerekmektedir. Gelir, üretim ve hizmet süreçleri sonucunda gerçek veya tüzel kişilerin elde ettikleri kazanım olarak ifade edilmektedir (Kubar, 2011: 228). Bu kazanım, daha çok parasal bir kazanım 73 Bengü Kayapalı Aslan \| Bengü Kayapalı Aslan ile ilişkilendirilse de gelirin nakdi olmayan formu da mevcuttur. Buna genel çerçevede gayri nakdi gelir denilmektedir. Bu gelir türü, “aynî gelir”, “emsal gelir”, “yan fayda” ve “öz tüketim” gibi aralarında küçük de olsa farklılıklar olan kavramlarla ifade açıklanabilmektedir (Rakıcı ve Aktuğ, 2020: 881). Mevzuatın gelir kavramına bakışı, “Gelir, bir gerçek kişinin bir takvim yılı içinde elde ettiği, tasarruf ve harcamasına kaynak teşkil eden her türlü kazanç ve iratların safi tutarıdır”2 şeklindedir. Bir başka perspektifte gelir, Fischer tarafından belirli bir zaman dilimi boyunca hizmetlerin akışı olarak tanımlanmaktadır (Hewett,1925: 239). Gelire yakın bir kavram olarak kazanç, gelirden farklı bir kavramı ifade etmektedir. Kazanç, istihdamdan kazanılan parayı ifade ederken, gelir, kazancın ötesindeki kazançları, sosyal yardımları ve emekli maaşları vb. dahil olmak üzere elde edilen toplam parayı ifade etmektedir (Healey, 2019). Gelirin belirli kişi veya gruplarca paylaşımını konu alan gelir dağılımı, gelir ve dağılım/paylaşım kavramlarından meydana gelmektedir. Bu kavramların doğru anlaşılması, gelir dağılımı kavramının doğru anlaşılmasını sağlayacaktır. Gelirin tanımı ve içeriği yukarıda anlatılmıştır. Gelirin dağılımı veya gelirin bölüşümü ise öncelikle bir ekonomide elde edilen gelirin, bu geliri elde edenler arasındaki bölüşümünü ifade etmektedir (Akın, 2015: 10). Gelir dağılımı aynı zamanda bir ülkedeki ekonominin yapısını, ekonomik büyüme hızı ve istihdam dağılımını üzerinden etkilemektedir (Karataş, 2019: 55). Buna yönelik olarak hükümetlerin gelir dağılımı politikaları, geliri fazla olandan az olana doğru geliri yeniden nasıl dağıtması gerektiği üzerinedir (Levy,t.y.). Bir ekonomide elde edilen gelirin, o ekonomideki kişiler arasında nasıl bölüşüldüğünü, bunun yanında ücret, faiz, rant, kâr gibi üretim faktörlerinin milli gelir içerisindeki payının ölçülmesi gelir dağılımının odağında yer almaktadır (Eğilmez, 2019). 2 193 Sayılı Gelir Vergisi Kanunu’nda Madde 1’de yer alan gelir kavramına ilişkin tanımdır. https://www.mevzuat.gov.tr/mevzuat?MevzuatNo=193&MevzuatTur=1&MevzuatTertip=4 Gelir Dağılımı Türleri Gelir dağılımı; fonksiyonel gelir dağılımı, sektörel gelir dağılımı, bölgesel gelir dağılımı ve kişisel gelir dağılımı başta olmak üzere çeşitli başlıklarda incelenmektedir (Uysal, 2007: 250-253). Buna genel itibariyle gelir dağılımı türleri denilmektedir. 2 193 Sayılı Gelir Vergisi Kanunu nda Madde 1 de yer alan gelir kavramına ilişkin tanımdır. https://www.mevzuat.gov.tr/mevzuat?MevzuatNo=193&MevzuatTur=1&MevzuatTertip=4 Kişisel Gelir Dağılımı Kişisel gelir dağılımı temelde bir ekonomide elde edilen gelirin yine o ekonomideki kişiler arasında ne şekilde dağıldığını konu almaktadır Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 74 (Eğilmez, 2019). Bir başka perspektifte kişisel gelir dağılımı, kaynak türlerine bakılmaksızın hanelerin veya bireylerin gelirlerinin sıklığı veya büyüklüğünün dağılımı olarak da ifade edilebilir (Kurt, 2018: 1). Fonksiyonel bağlamda kişisel gelir dağılımı ise toplumdaki en yüksek ve en düşük gelir düzeyine sahip grupların toplam gelirden aldıkları payların belirlenmesinde önemli bir gösterge olarak kabul edilmektedir (Destek, 2018: 6). Dolayısıyla kişisel gelir dağılımında, en yüksek ve en düşük gelire sahip gruplar arasındaki farklar ve buna yol açan dinamikler irdelenmektedir. Fonksiyonel Gelir Dağılımı Fonksiyonel gelir dağılımı, gayrisafi yurtiçi hasılada içerisindeki emek geliri olan maaş/ücret ve emek dışı gelirler olan kar, faiz, kira vb. üretim faktörlerinin bölüşümünü içermektedir. Bu sebeple fonksiyonel gelir dağılımı temelde gelir ile üretim faktörleri arasındaki ilişkiyi inceler (Boratav,1991: 26). Diğer bir ifadeyle üretim faktörlerinin milli gelirden aldıkları payları inceleyen fonksiyonel gelir dağılımı, gelirin emek gelirleri ile emek dışı gelirler arasındaki bölüşümü olarak da ifade edilmektedir (Strateji ve Bütçe Başkanlığı, 2001: 4). Sektörel Gelir Dağılımı Sektörel gelir dağılımı, öncelikle gayri safi yurtiçi hasılayı tarım, sanayi, ticaret, hizmet vb. ekonomideki üretim sektörlerine göre ayırmaktadır. Yıllara ayrılarak incelenebilen bu dağılım türü aynı zamanda ülkenin gelişmişlik düzeyi açısından bir gösterge olarak da kabul edilmektedir (Kuştepeli ve Halaç, 2004: 147). Çünkü ülkedeki sanayileşme sürecinde ülkenin mevcut konumu, üretim yapılan sektörler, sektörel gelişim planları ve sanayileşme politikalarında izlenmesi gereken strateji gibi birçok noktada önemli bir veriyi oluşturmaktadır. Lorenz Eğrisi Gelir dağılımı ölçüm yöntemlerinden birisi olarak Lorenz eğrisi, gayri safi yurtiçi hasılanın ülkedeki nüfus içindeki dağılımını gösteren bir ölçüm aracıdır. Bu ölçüm aracıyla incelenen bir ülkede bu ölçüm aracı mutlak eşitlik doğrusundan uzaklaştıkça gelir dağılımında eşitsizliğin arttığı anlaşılmaktadır. Lorenz eğrisinin yatay ve diken eksende oluşu ise tam eşitsizlik durumudur. Bu durumun tam aksine Millî gelirin bireyler arasında tamamıyla eşit dağılması durumunda Lorenz eğrisi, kare içindeki köşegenle temsil edilen mutlak eşitlik doğrusu ile çakışır (Bellù, 2005: 3). İstenilen ancak gerçekleşmeyen bu durum, gelir dağılımında eşitsizliğin olmadığı durumu ifade etmektedir. Bölgesel Gelir Dağılımı Bölgesel gelir dağılımı, bir ülkede farklı bölgelerdeki bireylerin milli gelirden ne ölçüde pay aldıklarını araştırmaktadır. Bir başka ifadeyle ülkedeki toplam gelirin bölgeler arasında bölüşümü ve bölgelerin milli gelirden ne ölçüde yararlanabildiğini konu edinir. Bu noktada ortaya çıkan farklılıklar bölgesel eşitsizliklerin de temelini oluşturur. Bölgeler arasında gelir dağılımında ortaya çıkan eşitsizlikler eğitimden sağlığa, sanayiden tarıma sosyoekonomik farkların oluşmasına neden olmaktadır (Aktan, 2002: 1). Dolayısıyla bu farklılıklar, bölgeler arasında gelir dağılımı eşitsizliğinin hem nedenini hem de sonucunu oluşturmaktadır. Bengü Kayapalı Aslan \| 75 75 Diğer Gelir Dağılımı Türleri Diğer Gelir Dağılımı Türleri Bir ekonomide milli gelirin üretim faktörlerince bölüşümünü irdeleyen bir kavram olarak gelir dağılımının bireysel, fonksiyonel, sektörel ve bölgesel gelir dağılımı haricinde başka türleri de vardır. Ancak literatürde yer alan çalışmalarda gelir dağılımının türü olarak bu dört başlığı kullanmakta olduğu görülmüştür. Gelir Dağılımı Eşitsizliği Ölçüm Yöntemleri Gelir Dağılımı Eşitsizliği Ölçüm Yöntemleri Gelir Dağılımını Etkileyen Başlıca Faktörler Gelirin büyüklüğü veya kaynağı yerine gelirin bölüşümünün önem taşıdığı perspektifte, David Ricardo da Malthus’a gönderdiği mektupta politik iktisadın öncelikli amacının refah artışı değil, bu refahın bölüşüm sorunu olduğunu belirtmiştir (Keynes,1936: 15). David Ricardo’nun 200 yılı aşkın süre öncesinde değindiği gelirin dağılımını; üretim faktörlerinin dağılımı, gelirin yeniden dağıtımı üzerine geliştirilen politikalar, küreselleşme, kamu gelirleri, işsizlik, teknoloji ve eğitim gibi faktörler başlıca etkilemektedir (Güder, 2019: 199). Gelir Dağılımı Eşitsizliği Ölçüm Yöntemleri Theil Endeksi Theil indeksi, gelir eşitsizliğini gruplar arası ve grup içi eşitsizlik olarak ayrıştırmaya imkan sağladığı için tercih edilen bir yöntemdir. Toplam eşitsizliği gruplara ayırabilmesi yönüyle ampirik araştırmalarda sıkça kullanılmaktadır (Akın ve Aytun, 2017: 56). Theil endeksi, bir ülkenin veya bölgenin gelir dağılımlarını karşılaştırılırken kullanılır daha çok kullanılmaktadır. Bu endekste zengin bireyden yoksul bireye doğru gelir aktarımı olduğunda eşitsizlik azalır ve endeks küçülür. Endeksin sıfıra eşit olduğu durum ise tam adil dağılım durumudur (Özdemir, 2017: 95). Gini katsayısı gelir eşitsizliği ölçümlerinde sıklıkla tercih edilen bir yöntem olmasına rağmen gelir dağılımının gelir dağılımının her iki ucuna duyarlı olan Theil endeksi de literatürde sıkça kullanılan bir yöntemdir. Gini Katsayısı Gini katsayısı, gelir eşitsizliğinin ölçümünde en yaygın şekilde kullanılan ölçüm yöntemlerinden birisidir. İtalyan istatistikçi Corrado Gini tarafından geliştirilmiştir. Gini katsayısı temelde toplam gelirler ile nüfustaki ortalama gelir arasındaki beklenen mutlak farkı ifade etmektedir (Hasell, 2023). Gelir dağılımındaki eşitsizlik derecesini ölçen “gini” katsayısı, 0 ile 1 arasında değer almaktadır. Gelirin tamamı bir kişiye ait olduğu durumda gini katsayısı 1 değerini alırken, gelir bireyler arasında eşit bölüşüldüğünde ise sıfır değerini almaktadır. Buna mutlak eşitlik durumu da denilmektedir. Dolayısıyla gini Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 76 \| 76 katsayısının değeri sıfıra yaklaştıkça eşitlik artarken değer 1’e yaklaştıkça gelir dağılımı eşitsizliği artmaktadır (Akın, 2015: 11) Atkinson Eşitsizlik Ölçütü Gelir dağılımına ilişkin hesaplamaların yapıldığı ölçüm yöntemlerinde odak, gelirin bölüşümü üzerinedir. Bunlardan farklı olarak Atkinson, sosyal refahın da üzerinde durulması gereken önemli bir konu olduğunu ifade etmektedir. Bir başka ifadeyle Atkinson, gelir dağılımı yaklaşımında sosyal refahı ve refah kaybına odaklanmaktadır (Bosmans ve Schokkaert, 2004: 100). Bu ölçümde diğer ölçümlerden farklı olarak hesaplama yapılırken, dağılımın üst veya alt kısmındaki gelir seviyelerine ağırlık verecek şekilde ağırlıklandırma değişkeni eklenmektedir (Akdağ, 2020: 34). Pareto Katsayısı Gelir eşitsizliği konusunda görece karamsar bir tablo çizen Pareto, gelir eşitsizliğinin bu durumu ortaya çıkaran güçlü ülkeler kadar değiştirilemeyecek, sürekli ve olağan bir tablo olarak görmektedir. Pareto’ya göre gelirin/kaynaklarn bölüşümünde bir kişinin refahını yükseltmek için diğer bir kişinin refahını düşürmek gerekiyorsa toplumun maksimum refah durumunda olduğu kabul edilmektedir (Akın, 2015: 14). Bir geliş eşitsizliği ölçümü olarak geliştirdiği yöntemde ise Pareto, gelirin bölüşümünde genel denge üzerine vurgu yapmaktadır (Pen, 1971: 235). Bu yaklaşımdan hareketle bu değer bir optimum olarak da kabul görmektedir. Merkantalistlerin ve Fizyokratların Gelir Dağılımı Yaklaşımı 18. yüzyılın ikinci yarısında Fransa’da gelişen Fizyokrasi sonucunda ortaya çıkmıştır. Quesnay Kuramı olarak da bilinmektedir. Fizyokratların iktisadi düşünce okulu, liberal iktisat düşüncesinin temelini oluşturmuştur (Bocutoğlu, 2012: 35). Fizyokratların yaklaşımında bölüşümün/paylaşımın temelinde tarım yer almaktadır. Çünkü tarım, fizyokrat düşüncede üretken olan yegâne sektör olarak kabul edilmektedir. Buna göre tarımda üretici konumdakiler, yani verimli sınıf, toprakta emek ve sermayeyi kullanarak üretim yapmaktadır. Ekonomik faaliyetler sürekli bir döngü halindedir ve bu döngü sonucunda elde edilen ürünler toplumdaki sınıflar arasında paylaşılmaktadır (Milanoviç, 2015: 18-22). Diğer Yöntemler Yukarıda ifade edilen ölçüm yöntemleri, gelir dağılımı eşitsizliğinin belirlenmesinde kullanılan yöntemlerden birkaçıdır. Bunların haricinde; Bengü Kayapalı Aslan \| 77 Bengü Kayapalı Aslan \| 77 Değişim Aralığı Yöntemi, Değişim Katsayısı, Göreli Ortalama Mutlak Sapma, Pearson Çarpıklık Katsayısı, Göreli Varyans, Logaritmik Standart Sapma, Kuznets Katsayısı, Elteto-Frigyes Eşitsizlik Ölçütü, Dalton Eşitsizlik Ölçütü, Sıklık Fonksiyonu ve Hareketlilik Matrisi gelir dağılımı eşitsizliğinin ölçümünde kullanılan yöntemleri ifade etmektedir. Değişim Aralığı Yöntemi, Değişim Katsayısı, Göreli Ortalama Mutlak Sapma, Pearson Çarpıklık Katsayısı, Göreli Varyans, Logaritmik Standart Sapma, Kuznets Katsayısı, Elteto-Frigyes Eşitsizlik Ölçütü, Dalton Eşitsizlik Ölçütü, Sıklık Fonksiyonu ve Hareketlilik Matrisi gelir dağılımı eşitsizliğinin ölçümünde kullanılan yöntemleri ifade etmektedir. İktisadi Ekollerin Gelir Dağılımı Yaklaşımları Gelir dağılımı, iktisadi ekollerce farklı perspektiflerde incelenmiştir. İktisadi ekollerin gelir dağılımına yaklaşımı, sahip oldukları iktisat teorisi çerçevesinde şekillenmiştir. Bunun sonucunda iktisat okulları kendilerine ait perspektifte gelir dağılımına ilişkin modeller geliştirerek konuyu ele almışlardır (Çiftçi, 2018: 22). Klasik İktisadın Gelir Dağılımı Yaklaşımı Klasik ekolün oluşmasında liberal felsefenin bir ürünü olarak anılan Adam Smith’in “Ulusların Zenginliği” isimli eseri yer almaktadır. Bunun yanında David Ricardo, Robert Malthus, Frédéric Bastiat ve J.B. Say klasik okulun oluşmasına önemli katkılar sağlamıştır. Bu ekolün gelir dağılımına yaklaşımının temellerinde D.Ricardo’nun çalışmaları yer almaktadır. Bu sebeple klasik okulun gelir dağılımı yaklaşımı Ricardo’nun azalan getiri teorisi üzerinedir. Ricardo’nun gelir dağılımı teorisinde ekonomi tarım ve sanayi olmak üzere iki kesime indirgenerek incelenir. Bunun yanında tarımın görece üstünlüğü olsa da ekonominin gelişimi konusunda üç ana varsayım sunmuştur. Bunlar; tarım sektörünün azalan getiriye sahip olduğu, ücret limitinin belirlenmesinde nüfusun belirleyiciliği ve son olarak da sermaye birikiminin iktisadi gelişmedeki belirleyiciliği üzerinedir (Ricardo,1997: 39-40). 78 \| Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme Keynesyen Gelir Dağılımı Yaklaşımı Gelir dağılımı, Keynes tarafından kapitalist bir ekonominin iki önemli sorunundan birisi olarak ifade edilmektedir (Keynes, 1935: 185). Keynes’in gelir dağılımı eşitsizliğine yaklaşımı iki varsayıma dayanmaktadır. Bunlar; • Ücretlerin parasal olarak belirlenmesi • Yatırımların, tasarruflardan bağımsız olarak belirlenmesidir. Çünkü Keynesyen yaklaşımda geliri oluşturan ve tasarrufun yapılmasını sağlayan temel faktör, yatırımlardır. Dolayısıyla Keynesyen gelir dağılımı yaklaşımında gelir dağılımının adil şekilde sağlanması tasarruf-yatırım eşitliğine bağlıdır (Destek, 2018: 12-13). Bunun yanında Keynesyen okulun gelir dağılımı yaklaşımında eşitsizliğin ortadan kaldırılmasına veya azaltılmasına dair politikalar doğrudan yer almamaktadır. Gelir dağılımı eşitsizliği toplumda kabul gören bir sorun olmakla birlikte Keynesyen yaklaşım, devlet müdahalesine karşı çıkmamakla liberal yaklaşımdan ayrışmaktadır. Bunun yanında devletin uygulayacağı iktisat politikalarının gelir dağılımı eşitsizliği sorununu çözebileceğine ve gelirin daha adil dağılacağını öngörmektedir. Neo-Klasik İktisadın Gelir Dağılımı Yaklaşımı Emek maliyeti yerine fayda değerinin alınması başta olmak üzere Marksist iktisatın savunularıyla tezatlıklar barındıran neo-klasik iktisat, klasik iktisat okulunun kabullerinin aksine gelir dağılımının ekonomide yer alan bir sorun olmadığını öne sürmüştür. Bunun temelinde herkesin üretim sürecinde yer aldığı ölçüde gelir ettiği yer almaktadır. Dolayısıyla hiçbir kimsenin elinde bir fazla olmayacağı için gelir adil şekilde dağılmış olacaktır (Ulutürk ve Ersezer, 2005: 94-95). Bu çıkarım hem klasik hem de neo-klasik iktisatın temel savunularından olan ekonomiye devlet müdahalesinin olmaması gerekliliği ile örtüşmektedir. Devlet müdahalesinin olmadığı ve piyasada kaynakların- yatırımların sektörlere ve mal üretimine dağılımı konusunda geçerli rekabet şartları, uzun dönemde normal üstü karları, normal karlara dönüştüreceği için gelir dağılımında düzenlenmeyi sağlanmış olacaktır (Birol ve Gencer, 2014: 262-266). 3 İktisat bilimi, David Ricardo tarafından “hasılayı üreten sınıflar arasında, gelir dağılımının yasalarının incelenmesi” olarak tanımlanmıştır. (bkz. Güneri Akalın: Kamu Ekonomisi, Sf.299) Diğer Yaklaşımlar Gelir dağılımındaki eşitsizliklerinin olası nedenleri, gelir dağılımı adaletinin sağlanmasına yönelik politikalar ve temel iktisadi görüşlerin gelirin paylaşımındaki yaklaşımları gelir dağılımına ilişkin yaklaşımlar olarak nitelendirilmiştir. Yukarıda sıklıkla başvurulan gelir dağılımı yaklaşımlara yer verilmiştir. Bu yaklaşımların yanında; • Neo-Keynesyen İktisadın Gelir Dağılımına Bakışı, • Post Keynesyen Teoride Gelir Dağılımı yaklaşımı, • Arz Yanlı İktisadın Gelir Dağılımına Bakışı, • Monetarist Okulun Gelir Dağılımına Bakışı, • Rasyonel Beklentiler Teorisinde Gelir Dağılımı gelir dağılımı konusundaki iktisadi yaklaşımlardan birkaçını ifade etmektedir. Marksist İktisadın Gelir Dağılımı Yaklaşımı Marksist iktisadın gelir dağılımı yaklaşımının temelinde emek-değer ve artık-değer ilkeleri yer almaktadır. Marks’a göre artık değerin kaynağı emektir. Bu sebeple David Ricardo’ nun artık-değer ilkesinden bu noktada ayrılmaktadır. Bunun yanında Marksist yaklaşımda yine Ricardo’nun aksine 79 Bengü Kayapalı Aslan \| 79 Bengü Kayapalı Aslan \| Bengü Kayapalı Aslan \| azalan verim prensibi kabul edilmemektedir. Ayıca rant ile kâr arasında da ayrımın olmadığı savunulmaktadır (Akalın, 1986: 305-306). Kapitalist ekonomilerde sanayileşme ve teknolojik gelişmelere bağlı olarak ortaya çıkacak işgücü fazlası emek arzında fazlalık oluşturacağı için asgari geçim düzeyinin düşmesine neden olacaktır. Marks, bu durumu işçi sınıfının sefaleti olarak tanımlamaktadır. Buradan anlaşılacağı üzere Marksist teorinin gelir dağılımı noktasındaki yaklaşımı; gelirin bölüşümü-emek-sermaye çatışması üzerine kurulmuştur (Kazgan, 2006: 311-313). Gelir Dağılımına İlişkin Literatürün İncelenmesi İktisat Bilimi hakkında en bilinen tanımlar arasında yer alan gelir dağılımı3, önemli bir çalışma konusunu ifade etmektedir. Bu noktada gelir dağılımına ilişkin yapılan çalışmalar irdelendiğinde; 0 \| Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 80 Tablo 1. Gelir Dağılımının Temel Dinamiklerine Yönelik Literatür Özeti Tablo 1. Gelir Dağılımının Temel Dinamiklerine Yönelik Literatür Özeti Kaynak : (Destek, 2018: 31). 81 Bengü Kayapalı Aslan \| 81 Sonuç ve Öneriler Dünya’da küresel iklim değişikliği, azalan doğal kaynaklar, pandemi düzeyinde salgınlar ve teknolojik gelişmelerin yanında diğer önemli bir konu gelir dağılımındaki adalet sorunudur. Etkisinin her geçen gün daha da fazla hissedilen bu kavram, derinlemesine incelenmesi gereken sosyoekonomik bir gerçektir. Yoksulluğun ve gelirin bölüşümündeki eşitsizliğe bağlı olarak artan sorunlar, gelir dağılımı konusunun önemine vurgu yapmaktadır. Yoksulluğun azaltılarak insan yaşamının standartlarının iyileştirilmesi ve bunlara yönelik politikaların oluşturulması hem ulusal hem de uluslararası düzeyde ülkelerin öncelikli hedefleri arasında yer almaktadır. Birleşmiş Milletler tarafından deklare edilen “sürdürülebilir kalkınma amaçları” arasında da birçok hedef, alt başlık ve done ile ilişkilendirilebilecek gelir dağılımı kavramı, üzerine farklı bilim dalları ve disiplinlerce çalışmalar yapılan sürdürülebilir bir konuyu içermektedir. Bu noktada çalışmanın amacı öncelikli olarak gelir dağılımı kavramının tüm boyutlarıyla ele alınarak geniş bir perspektifte incelenmesidir. Çalışmada gelir dağılımı kavramı; gelir dağılımı türleri, kendisini etkileyen faktörler, gelir dağılımı ölçüm yöntemleri ve iktisadi ekollerin gelir dağılımına yaklaşımları başlıklarıyla detaylı şekilde incelenmiştir. Gelir dağılımına ilişkin halihazırda yapılan çalışmalara yer verilen literatürde özet nitelikte bir tablodan yararlanılmıştır. Tüm bunlardan hareketle çalışmada elde edilen öncelikli sonuç, artan gelir adaletsizliği tablosunda devletlerin gelir eşitsizliğini azaltıcı politikalar geliştirmesidir. Gelir dağılımı eşitsizliğine yönelik politikalar oluşturulurken yararlanılması gereken dayanak çalışmada da incelenen gelir dağılımının belirleyicileri üzerinedir. Buradan hareketle sunulacak öncelikli öneriler; • Gelir dağılımını inceleyen çalışmaların yeni endeks ve göstergelerle birlikte konuyu incelemeleri, • Devlet tarafından uygulanacak ekonomik ve sosyal politikaların gelir dağılımına etkilerinin doğru şekilde belirlenmesi, • Teknolojik gelişmelerden ve teşvik içerikli programlardan görece az gelişmiş bölgelerin daha fazla yararlanmasını sağlamak, • Beslenme, barınma, sağlık, eğitim, çevre ve kültür seviyesinde kötü koşullara sahip bölgelerin sürdürülebilir şekilde desteklenmesi, • Gelir dağılımı adaletinin sağlanmasında fırsat eşitliğinin gözetilmesi, • Dezavantajlı ve kırılgan grupların istihdama katılımının sağlanması, • Genel reçeteler yerine ülkelerin/bölgelerin gerçekliklerine uygun çözümlerin üretilmesi ve desteklenmesi şeklinde sıralanmaktadır. şeklinde sıralanmaktadır. şeklinde sıralanmaktadır. 82 \| Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme Kaynakça Akalın, G. (1986). 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Bir İktisat Politikası Amacı Olarak Gelir Dağılımı: Türkiye Örneği (1994–2007) Analizi. Kahramanmaraş Sütçü İmam Üniversitesi İktisadi ve İdari Bilimler Fakültesi Dergisi, 1 (2), 227-246. Retrieved from http://iibfdergisi.ksu.edu.tr/tr/pub/issue/10263/125881 Kurt, O.E. (2018). Personal And Functional Distributions Of Income in Turkey, Department of Economics, Notes on Economy, No:2018-03 Kuştepeli, Y. & Halaç, U. (2004). “Türkiye’de Genel Gelir Dağılımının Analizi ve İyileştirilmesi”, Dokuz Eylül Üniversitesi SBE Dergisi, 6(4), 143-160. Levy, F. (t.y.). Distribution of Income, https://www.econlib.org/library/Enc/ DistributionofIncome.html, Erişim Tarihi: 13.05.2023. Milanovic, B. (2015). The Level And Distribution Of Income In Mid-E- ighteenth-Century France, According To Françoıs Quesnay. Jour- nal of the History of Economic Thought, 37(1), 17-37. doi:10.1017/ S1053837214000728 Özdemir, M. Ç. (2017) ‘‘Gelir Eşitsizliği Ölçüm Yöntemleri. E. İslamoğlu ve M. Ç. Özdemir içinde, Gelir Dağılımı ve Yoksulluk’’: Sf. 78-110, Ankara: Seçkin Yayıncılık. Pen, J. (1971). Income Distribution, Facts, Theories, Policies. Wesport: Praeger Publishers. \| Gelir Dağılımı Kavramı ve Gelir Dağılımına İlişkin Literatür Üzerine Bir İnceleme 84 Rakıcı, C. & Aktuğ, M. (2020). Gelir Kavramına Yönelik Farklı Görüşler ve Türk Vergi Sisteminin Gelire Yaklaşımı. Bolu Abant İzzet Baysal Üniversi- tesi Sosyal Bilimler Enstitüsü Dergisi, 20 (4), 879-904. DOI: 10.11616/ basbed.v20i58861.838905 Ricardo D. (1997). Ekonomi Politiğin ve Vergilendirmenin İlkeleri, Çeviren: Tayfun Ertan, Belge Yayınları, İstanbul. Strateji ve Bütçe Başkanlığı (2001). Gelir Dağılımının İyileştirilmesi Ve Yoksul- lukla Mücadele Özel İhtisas Komisyonu Raporu, https://www.sbb.gov.tr/ wp-content/uploads/2022/08/Gelir-Dagiliminin-Iyilestirilmesi-ve-Yok- sullukla-Mucadele-OIK-Raporu.pdf, Erişim Tarihi: 17.05.2023. Ulutürk, S. & Ersezer, D. (2005). Gelir, Gelir Dağılımı Yaklaşımları ve Devletin Rolü. Maliye Araştırma Merkezi Konferansları, 0 (47), 87-105. Retrie- ved from https://dergipark.org.tr/tr/pub/iumamk/issue/745/8040 Uysal, Y. (2007). Gelir Dağılımı Türleri Arasındaki İlişkiler Perspektifinde Tür- kiye’de Gelir Dağılımının Düzenlenmesine Yönelik Öneriler, Dokuz Ey- lül Üniversitesi Sosyal Bilimler Enstitüsü Dergisi, 9(2), 248-292. Bölüm 6 Özet Ülkelerin ekonomik büyüme hedeflerini gerçekleştirebilmesi adına enerji kaynaklarının kullanımı büyük önem arz etmektedir. Ancak yenilenemeyen enerji kaynaklarının kullanımı ekonomik büyümeyi sağlarken diğer yandan çevre kirliliğine neden olarak ekolojik dengenin bozulmasına yol açmaktadır. Özellikle gelişmekte olan ülkeler tarafından en az maliyetle en yüksek çıktı hedeflemesi yapılmaktadır. Bu nedenle de sıklıkla fosil yakıtların tercih edildiği görülmektedir. Yenilenebilir enerji kaynakları ise sürdürülebilir enerjinin geleceği için en önemli dinamiklerden biridir. Dolayısıyla özellikle yeni sanayileşen ülkelerin yenilenebilir enerji kaynaklarına teşvik edilebilmesi açısından söz konusu bu kaynakların ekonomik büyüme üzerindeki etkisinin belirlenmesi gerekmektedir. Sera gazlarının küresel ısınmanın tetikleyicisi olduğunun anlaşılmasıyla birlikte çevre kirliliği ülkelerin gündem başlıkları arasındaki yerini almıştır. Bu doğrultuda da sürdürülebilir kalkınma hedefleri ortaya konulmuştur. Ancak sürdürülebilir kalkınma hedeflerinin birbiriyle çelişebildiğine dair görüşler de mevcuttur. Bu çalışmanın amacı 1990- 2015 gözlem aralığı baz alınarak Türkiye için sürdürülebilir kalkınma hedefleri kapsamında yer alan SDG 7 ve SDG 8 arasındaki muhtemel uyum ya da çatışmayı incelemektir. Ampirik modele dahil edilen değişkenler 2015 sabit fiyatları ile dolar cinsinden kişi başına düşen gayri safi yurtiçi hasıla, yenilenebilir enerji tüketimi (toplam nihai enerji tüketiminin yüzdesi), fosil yakıt enerji tüketimi (toplamın yüzdesi), brüt sabit sermaye oluşumunun gayri safi yurtiçi hasıla yüzdesi, işgücüne katılım oranı, 15-64 yaş arası toplam nüfusun yüzdesidir. Değişkenler arasındaki ilişki ARDL sınır testi ile araştırılmıştır. Serilerin durağanlık durumu ADF birim kök testi ile incelenmiştir. Elde edilen bulgular doğrultusunda yenilenebilir enerji kullanımının ekonomik büyümeye olumlu olarak katkı sağladığı saptanmıştır. Ayrıca söz konusu hedeflerin birbiriyle uyumlu olduğu belirlenmiştir. https://doi.org/10.58830/ozgur.pub401.c1754 https://doi.org/10.58830/ozgur.pub401.c1754 85 85 86 \| Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin Ekonomik Büyüme... 1.Giriş Ekonomik büyüme ülkelerin tartışmasız en temel amaçları arasında yer almaktadır. Ekonomik büyümenin gerçekleştirilebilmesi ise üretim miktarı ile doğru orantılıdır. Bu nedenle ülkeler en az maliyetle en yüksek çıktıyı hedeflemektedir. Tüketim ihtiyaçlarını hızlı bir şekilde karşılayabilmek özellikle kitlesel üretimlerin artması sayesinde mümkün hale gelebilmiştir. Kitlesel üretimler ise teknoloji ve enerji kaynaklarının bir arada kullanımıyla mümkün olmaktadır. Bu sayede çıktı oranlarında ciddi artışlar meydana gelmekte ve ekonomik büyüme oranlarında da artış sağlanabilmektedir. Tabi ki tüm bu süreç için de enerji kullanımı büyük önem arz etmektedir. Zira enerji üretim aşamasının hem girdisi hem de çıktısı konumunda yer almaktadır (Zou vd., 2016). Özellikle küreselleşmeyle beraber artan üretim faaliyetlerine paralel olarak enerji kullanımı da artmaktadır. Bu durum ise ülkelerin enerjiye olan ihtiyacının artmasına neden olmaktadır (Jiang vd., 2023). Ortaya çıkan enerji ihtiyacının ise yenilenemeyen enerji kaynakları ile karşılanması neticesinde çevre kirliliğine yol açarak ekosisteme zarar vermekte ve küresel ısınma tehlikesi ile karşı karşıya kalınmaktadır. Ayrıca fosil yakıt rezervleri yenilenemeyen enerji kaynakları arasında yer aldığı için bir gün tükenme ihtimalini de bünyesinde barındırmaktadır (Solomon vd., 2022). Fosil yakıtların yaygın olarak kullanılmasının temel nedeni ise yenilenebilir enerji kaynaklarının yenilenemeyen enerji kaynaklarına göre daha maliyetli olmasıdır (Kabeyi ve Olanrewaju, 2022). Ülkelerin yenilenebilir enerji kaynaklarına geçmesi ise ilk olarak 1973 petrol kriziyle gündeme gelmiştir. Söz konusu bu krizle birlikte fosil yakıtların sınırsız olmadığı anlaşılmış ve söz konusu bu yakıtlara karşı temkinli yaklaşılması durumu gündeme gelmiştir. Ayrıca çevre kirliliğinin küresel boyutta bir sorun haline dönüşmesi de yenilenebilir enerji kaynaklarına yönelmeyi tetikleyen bir diğer unsurdur (Basosi vd., 2020). Yenilenebilir enerji kaynakları arasında biyokütle enerji, rüzgâr enerjisi, dalga enerjisi, hidroelektrik enerji, jeotermal enerji ve güneş enerjisi gibi enerji türleri yer almaktadır (Moriarty ve Honnery vd., 2019). Çevre kirliliğindeki artışın ekonomik büyüme faaliyetlerinden dolayı kaynaklanmasından ötürü araştırmacıların da ilgisini çekmiş ve söz konusu iki unsur birbiriyle ilişkilendirilmeye başlanmıştır. Ekonomik büyüme ile çevre kirliliği arasındaki ilişki Kuznets Eğrisi hipotezine dayanmaktadır. Kuznets Eğrisi hipotezine göre ekonomik büyüme ile gelir eşitsizliği arasında ters U şeklinde bir ilişki bulunmaktadır (Kuznets, 1955). Daha sonraları ise Kuznets Eğrisi hipotezi, Çevresel Kuznets Eğrisi (EKC) hipotezine evrilmiştir. Grossman ve Krueger (1991), Panayotou (1993) ve Shafik’e (1994) dayanan EKC hipotezine göre, ekonomik büyümenin ilk başlarında çevresel bozulma artmaktadır. Ancak ekonomik 87 Elif Koçak Elif Koçak 87 büyümenin ilerleyen aşamalarında teknik ve yapısal etkilerin sayesinde çevre kalitesinde artış görülmektedir. Çevre kirliliğinin bölgesel olmayıp küresel bir sorun olmasının anlaşılmasıyla birlikte, sürdürülebilir bir çevre için ülkeler birtakım önlemler almaya başlamıştır. 1.Giriş Bu önlemlerin başında sürdürülebilir kalkınma planları gelmektedir. Sürdürülebilir kalkınmanın sosyal, ekonomik ve çevresel olmak üzere üç temel konsept etrafında şekillendiği söylenilebilir. Bu konsept dahilinde sürdürülebilir kalkınma hedefleri yoksulluğu, gelir eşitsizliğini ve çevresel bozulmayı arttırmadan gerçekleştirilecek bir ekonomik büyüme modeline vurgu yapmaktadır. Ekonomik olarak mal ve hizmetin kesintisiz olarak üretilebilmesi, dış borcun korunması gibi önemli faktörleri bünyesinde barındırmaktadır. Çevresel kısmı ekonomik büyümenin sağlanırken ekolojik dengeye zarar verilmemesi ve bunun için de yenilenemeyen enerji kaynaklarından kaçınmak gerektiğini ifade etmektedir. Sosyal kısmı ise gelir eşitliği, sağlık, eğitim vs. gibi faktörleri temsil etmektedir (Harris, 2000). Ancak sosyal, ekonomik ve çevresel boyutları arasında bazı ikilemlerin bulunduğu görülmektedir. Çevre kalitesinin düşmesinde en etkili olan faktörün ekonomik büyüme olduğu kabul edilmesine rağmen, ülkelerin ekonomik büyüme hedeflerinden vazgeçmedikleri görülmektedir. Bir yandan ekonomik büyüme gerçekleştirilirken diğer yandan bilinçsiz bir şekilde kullanılan fosil yakıtlar çevre kirliliğine neden olmaktadır (Destek, 2019). Sürdürülebilir bir çevre için temiz enerji kullanımı ve ekonomik büyüme iki farklı hedeftir. Ancak bu hedeflerin çelişip çelişmeyeceği konusunda farklı görüşler mevcuttur. Sürdürülebilir kalkınma hedeflerinden birine ulaşırken diğer hedefin bozulması durumu söz konusu hedeflerden istenilen verimin alınamamasına neden olacaktır. Bu nedenle sürdürülebilir kalkınma hedefleri bağlamında SDG7 ve SDG8 arasındaki muhtemel uyum ya da çatışmanın incelenmesi önem taşımaktadır. Bu doğrultuda gelişmekte olan ve yeni sanayileşen ülkelerden biri olan Türkiye için yenilenebilir enerji kaynakları kullanımının ekonomik büyüme üzerindeki etkileri incelenecektir. 2.Literatür Özeti Jebli ve Youssef (2015), 1980 ile 2010 yılları arasındaki dönemde 69 ülkeyi baz alarak, hem yenilenebilir hem de yenilenemeyen enerji kaynaklarından elde edilen enerji üretim-tüketim ile uluslararası ticaret arasındaki ilişkiyi incelemişlerdir. Çalışmanın sonuçlarına göre, üretim ile ticaret arasında iki yönlü bir nedensellik olduğuna dair göstergeler içermektedir. Bu sonuçlar, ticaretteki herhangi bir değişikliğin ekonomik sonucu ve de buna bağlı olarak üretimdeki değişikliklerin de ticareti etkileyebileceğini göstermektedir. Kısa vadede GSYİH ile yeşil enerji tüketimi arasında herhangi bir bağlantı 88 Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin Ekonomik Büyüme... bildirilmemiştir. Ancak değişkenler arasında uzun dönemli iki yönlü bir ilişki bulunmaktadır. Ocal ve Aslan (2013), YEK ile GSYİH arasındaki ilişkiyi Türkiye için 1990-2010 gözlem aralığındaki verileri kullanarak gerçekleştirdiği çalışmada YEK’in GSYİH üzerinde negatif etkisi olduğunu ve ekonomik büyümeden yenilenebilir enerjiye nedensellik ilişkisi olduğu ortaya koyulmuştur. Fang (2011), YEK tüketiminin Çin’in GSYİH’si üzerindeki etkisini incelemiştir. Regresyon teknikleri kullanılarak yapılan analizde, YEK tüketimindeki %1’lik bir artışın %0,162’lik bir GSYİH büyümesine, kırsal haneler için %0,44’e ve kentsel haneler için %0,368’e yol açtığı sonucuna ulaşmıştır. Ancak YEK payının artmasının kaçınılmaz olarak politik- ekonomik engeller doğuracağı öne sürülmüştür. Fosil yakıt kullanımının insan sağlığının bozulması, altyapı, ormanların ve balıkçılığın azalması ve iklim değişikliğiyle mücadeleyle ilişkili harcamaların artması gibi toplum için maliyetleri içeren çevresel etkilerine atfedilmektedir. Tüm dış ekonomiler dikkate alındığında bazı YEK’lerin, özellikle de rüzgâr enerjisinin, yenilenemeyen enerji kaynaklarından daha ucuz olduğunun kanıtlandığı belirtilmiştir. Siyasi engellerle ilgili olarak, koordinasyon eksikliği, destekleyici olmayan politika teşviklerinin yanı sıra coğrafi kapsam veya sağlam araştırma analiz bağlamları açısından yeterince geliştirilmemiş devlet hibeleri rapor edilmiştir. Rafindadi ve Öztürk (2017), Almanya için 1971-2013 dönem aralığını kapsayan çalışmasında ARDL sınır testi ile Bayer-Hanck eşbütünleşme analizleri uygulayarak YEK ile GSYİH arasındaki ilişkiyi araştırmıştır. Sonuçlara göre çift yönlü nedensellik saptanmış olup, YEK’in GYSİH’yi pozitif yönde etkilediği saptanmıştır. Salim vd., (2014), 1980–2011 dönemi için OECD ülkelerine ilişkin çalışma gerçekleştirmiştir. Çalışmanın bulgularına göre hem yeşil hem de fosil enerji tüketiminin endüstriyel üretim ve ekonomik büyümeyle ilişkilendiren uzun vadeli bir ilişkiyi ortaya koymuştur. Ayrıca, kısa vadede fosil enerji tüketimi ile GSYİH büyümesi arasında çift yönlü bir ilişkinin olduğuna dair kanıtlar sunulmuştur. Bu sonuçlara göre, YEK’lerin genişletilmesi enerji güvenliği ve iklim değişikliğiyle mücadelede geçerli bir çözüm iken, fosil yakıtların kademeli olarak YEK’lerle değiştirilmesi sürdürülebilir bir enerji ekonomisini teşvik etmektedir. Farhani (2013), 12 MENA ülkesi için 1975-2008 dönemine ait verilerle Granger Nedensellik testi yöntemini kullanarak YEK ile GSYİH arasındaki ilişkiyi araştırmıştır. GDPt=β0+ β11nRt+ β2lnFt+ β3lnKt+ β4lnLt+ β5lnPt +μt (1) (1) Modelde GDP, 2015 sabit fiyatları ile dolar cinsinden kişi başına düşen gayri safi yurtiçi hasılayı; R, yenilenebilir enerji tüketimi (toplam nihai enerji tüketiminin yüzdesi); F, fosil yakıt enerji tüketimi (toplamın yüzdesi); K, brüt sabit sermaye oluşumu (GSYİH’nin yüzdesi); L, işgücüne katılım oranı; toplam (15-64 yaş arası toplam nüfusun yüzdesi); P, toplam nüfusu ve μt hata terimini ifade etmektedir. Ayrıca tüm değişkenler logaritmik formda kullanılmıştır. Çalışmada serilerin durağanlık durumu ADF birim kök testi ile araştırılmıştır. Değişkenler arasındaki eşbütünleşme ilişkisi için ise ARDL sınır testi uygulanmıştır. Son olarak, ARDL katsayı tahmincisi ile kısa ve uzun dönem katsayı tahmini yapılmıştır. 2.Literatür Özeti Söz konusu ampirik çalışmanın sonuçlarına göre YEK 89 Elif Koçak ile GSYİH arasında kısa dönemde nedensellik ilişkisi saptanmışken, uzun dönemde nedensellik ilişkisi saptanamamıştır. Apergis ve Payne (2012), 80 ülkenin verileri ile gerçekleştirmiş oldukları çalışmanın sonuçlarına göre; YEK tüketimi, yenilenemeyen enerji ve GSYİH arasında hem kısa hem de uzun vadede iki yönlü bir ilişki olduğunu ortaya koymuştur. Ayrıca yenilenebilir ve yenilenemeyen enerji türleri arasında iki yönlü kısa vadeli bir ilişki bulunduğu ve dolayısıyla enerji türleri arasında geçişin mümkün olduğu anlamına geldiği ifade edilmiştir. Sarı ve Soytaş (2004), farklı enerji tüketiminin reel çıktı üzerinde farklı etkileri olduğunu; fosil yakıtların da kullanıldığı Türkiye’de ekonomik kalkınma açısından enerji tüketiminin neredeyse istihdam kadar önemli göründüğü sonucuna ulaşmışlardır. Doğan ve Akçiçek (2013), Türkiye için 1980-2013 dönem aralığı verilerini baz alarak gerçekleştirmiş oldukları çalışmalarında Granger Nedensellik testini kullanarak YEK ile ekonomik büyüme arasındaki ilişkiyi incelemişlerdir. Söz konusu çalışmanın sonucuna göre YEK ile GSYİH arasında çift yönlü nedensellik ilişkisi saptanmıştır. Bhattacharya vd., (2016); biyokütle, jeotermal enerji, rüzgâr enerjisi, güneş enerjisi ve hidroelektrik enerjisi dahil olmak üzere YEK’lerin ekonomik büyüme üzerindeki etkisi, üç grupta sınıflandırılan 38 ülke için incelemiştir. Elde edilen sonuçlara göre birinci grupta yer alan 24 ülke için YEK’ler ekonomik büyümeyle önemli ölçüde ilişkilidir. İkinci gruba giren ülkelerde ise enerji tüketimi ekonomik kalkınmayla negatif ilişkilidir. Üçüncü grupta yer alan diğer ülkeler için YEK tüketiminin ekonomik büyüme üzerinde herhangi bir etkisi bulunmamaktadır. Shahbaz vd., (2015), YEK ile GSYİH arasındaki ilişkiyi 1972Q1- 2011Q4 dönemine ait 3 aylık verilerini kullanarak Pakistan için incelemiştir. Çalışmanın bulgularına göre YEK ile GSYİH arasında uzun dönemli eşbütünleşme ilişkisi ve çift yönlü nedensellik ilişkisi tespit edilmiştir. Inglesi-Lotz (2016), panel verileri ile gerçekleştirdiği çalışmasında YEK tüketimi, incelenen ülkelerin toplam kullanılan enerjinin YEK yüzdesi, iş gücü, araştırma ve geliştirme maliyetleri ve sermaye değişkenlerini kullanarak gerçekleştirdiği çalışmasında YEK tüketimindeki %1’lik bir artışın toplam GSYİH’yi %0,105 artırabildiğini, bir ülkenin enerji karışımında YEK’lerin payındaki %1’lik bir artışın ise toplam GSYİH’yi %0,089 artırabildiği sonucuna ulaşmıştır. 90 \| Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin Ekonomik Büyüme... 3.Model, Veri ve Yöntem Bu çalışmada 1990-2015 gözlem aralığı baz alınarak Türkiye için yenilenebilir enerjinin ekonomik büyüme üzerindeki etkilerinin incelenmesi amaçlanmıştır. Bu doğrultuda 2015 sabit fiyatları ile dolar cinsinden kişi başına düşen gayri safi yurtiçi hasıla, yenilenebilir enerji tüketimi (toplam nihai enerji tüketiminin yüzdesi), fosil yakıt enerji tüketimi (toplamın yüzdesi), brüt sabit sermaye oluşumu (GSYİH’nin yüzdesi), işgücüne katılım oranı ve 15-64 yaş arası toplam nüfusun yüzdesi verileri kullanılmıştır. Veriler “World Development Indicators” veri tabanından elde edilmiş olup, oluşturulan model aşağıdaki gibidir: GDPt=β0+ β11nRt+ β2lnFt+ β3lnKt+ β4lnLt+ β5lnPt +μt (1) 4. Ampirik Sonuçlar Elif Koçak \| 93 Elif Koçak \| 93 Grafik 1: CUSUM ve CUSUMQ Testleri -8 -6 -4 -2 0 2 4 6 8 2010 2011 2012 2013 2014 2015 CUSUM 5% Significance -0.4 0.0 0.4 0.8 1.2 1.6 01 02 03 04 05 06 07 08 09 10 11 12 13 14 15 CUSUM of Squares 5% Significance Tablo 3’te yer alan sonuçlar doğrultusunda kısa dönemde YEK’in GSYİH’ye anlamlı bir katkısı olmazken, uzun dönemde beklenildiği üzere GSYİH’yi artırdığı görülmektedir. Bu bulgu biyoenerji, rüzgâr enerjisi, hidroelektrik enerji, jeotermal enerji ve güneş enerjisi gibi yenilenebilir enerji kaynakları kullanımının ekonomik büyüme üzerindeki olumlu etkisinin kanıtıdır. Benzer şekilde fosil yakıtların kısa dönemde GSYİH’ye anlamlı bir katkısı bulunmazken, uzun dönemde GSYİH’yi artırdığı saptanmıştır. Bu durum üretim esnasında halen fosil yakıtların önemli bir yer tuttuğuna işaret etmektedir. Petrol, doğalgaz ve kömür gibi yenilenemeyen enerji kaynakları ekonomik büyümenin itici güçleri arasında yer almaktadır. Bu nedenle özellikle yeni sanayileşen ülkelerde CO2 salınımı kaçınılmaz olmaktadır. Sermayenin her iki dönemde de beklenildiği üzere GYSİH’yi artırdığı tespit edilmiştir. Emeğin kısa dönemde GSYİH’ye anlamlı bir katkısı olmazken, uzun dönemde GSYİH’yi artırmaktadır. Bu bulgu 15-64 yaş arası çalışan nüfusun ekonomik büyüme için önemli bir faktör olduğunun kanıtıdır. Nüfus artışının kısa dönemde GSYİH üzerinde olumsuz etkisi mevcutken, uzun dönemde GSYİH’ye olumlu etkisinin olduğu görülmektedir. Bu bulgu gençlerin üretken hale gelip ekonomiye katkı sağladığına işaret etmektedir. Son olarak, beklenildiği gibi hata düzeltme katsayısının negatif ve istatistiksel olarak anlamlı olduğu görülmektedir. 4. Ampirik Sonuçlar Yenilenebilir enerji kaynaklarının ekonomik büyüme üzerindeki etkilerinin incelendiği bu çalışmada ilk olarak serilerin birim kök içerip içermediğinin araştırılması gerekmektedir. Bunun için ADF birim kök testi uygulanmıştır. Tablo 1’de yer alan bulgulara göre düzey değerlerinde null hipotezin reddedilemediği ancak fark değerlerinde null hipotezinin reddedilerek serilerin birim kök içermediği görülmektedir. Elif Koçak \| 91 Elif Koçak \| 91 Tablo 1: ADF Birim Kök Testi Düzey Değerleri t-istatistiği Olasılık lnGDP 0.492129 0.9829 lnR -1.124920 0.6894 lnF -1.363934 0.5832 lnK -1.735374 0.4021 lnL -2.259924 0.1930 lnP 3.079637 1.0000 Fark Değerleri t-istatistiği Olasılık dlnGDP -5.004876* 0.0005 dlnR -6.403561* 0.0000 dlnF -4.508044* 0.0017 dlnK -5.244311* 0.0003 dlnL -4.616845* 0.0013 dlnP -3.435080 0.0277 Not: ,, sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. Tablo 1: ADF Birim Kök Testi Not: *,,* sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. Birim kök testinin ardından seriler arasında eşbütünleşme ilişkisinin varlığının sınanması gerekmektedir. Bunun için ARDL sınır testi uygulanmıştır. Tablo 2’de sunulan bulgulara göre modelin f-istatistiğinin I(0) ve I(1) bantlarının üzerinde olduğu görülmektedir. Bu durum ise seriler arasında bir eşbütünleşme ilişkisinin varlığını göstermektedir. Tablo 2: ARDL Sınır Testi Sonuçları Model Gecikme Uzunlukları f-istatistiği GDPt=f( Rt, Ft, Kt, Lt, Pt) (2, 2, 2, 2, 2, 2) 11.51459 Kritik Değerler I(0) I(1) %1 %5 %10 2.08 2.39 3.06 3 3.38 4.15 Not: *,,* sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. Tablo 2: ARDL Sınır Testi Sonuçları Not: *,,* sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. Eşbütünleşme ilişkisinin varlığının tespit edilmesinden sonra katsayı tahmini aşamasına geçilebilmektedir. Ancak öncelikle modelin güvenilirliğinin test edilmesi gerekmektedir. Tablo 3’te yer alan sonuçlara göre modelde otokorelasyon sorunun olmadığı Breusch-Godfrey (LM) testi ile, değişen varyans sorunun olmadığı ARCH testi ile, serilerin normal dağılım koşulunu 92 \| Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin Ekonomik Büyüme... 92 sağladığı, Jarque-Bera (JB) testi ile ve modelde doğru fonksiyonel formun kullanıldığı Ramsey-Reset testi ile tespit edilmiştir. Ayrıca CUSUM ve CUSUMQ testleri uygulanarak modelde parametrelerin istikrarlı oldukları teşhis edilmiştir. Tablo 3: ARDL Kısa ve Uzun Dönem Katsayı Tahmin Sonuçları Bağımlı Değişken: GDP Kısa Dönem Katsayı Olasılık R -0.022349 0.7187 F -0.763949 0.1330 K 0.392491* 0.0000 L -0.465380* 0.0006 P -3.033660* 0.0058 ECT(-1) -1.374413* 0.0000 Uzun Dönem R 0.764624* 0.0045 F 5.849592* 0.0029 K 0.583310* 0.0000 L 0.514142* 0.0073 P 1.748315* 0.0000 Varsayım Testleri ARCH 1.259327 0.3065 JB 3.076118 0.214798 LM 5.164825 0.3217 RAMSEY 0.815597 0.4518 CUSUM İstikrarlı İstikrarlı CUSUMQ İstikrarlı İstikrarlı Not: ,, sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. Tablo 3: ARDL Kısa ve Uzun Dönem Katsayı Tahmin Sonuçları Not: *,,* sırasıyla %1,%5,%10 anlamlılık düzeyini göstermektedir. 5.Sonuç Bu çalışmada sürdürülebilir kalkınma hedefleri arasında yer alan SDG 7 ve SDG 8 arasındaki muhtemel uyum ya da çatışmanın incelenmesi amaçlanmıştır. Bu doğrultuda Türkiye için 1990-2015 dönem aralığı baz alınarak yenilenebilir enerjinin ekonomik büyüme üzerindeki etkisi incelenmiştir. Oluşturulan ampirik model için 2015 sabit fiyatları ile dolar cinsinden kişi başına düşen gayri safi yurtiçi hasıla, yenilenebilir 94 Sürdürülebilir Kalkınma Hedefleri Doğrultusunda Yenilenebilir Enerjinin Ekonomik Büyüme... enerji tüketimi (toplam nihai enerji tüketiminin yüzdesi), fosil yakıt enerji tüketimi (toplamın yüzdesi), brüt sabit sermaye oluşumu (GSYİH’nin yüzdesi), işgücüne katılım oranı, 15-64 yaş arası toplam nüfusun yüzdesi verileri kullanılmıştır. Değişkenler arasındaki ilişki ise ARDL sınır testi ile araştırılmıştır. Elde edilen bulgulara göre yenilenebilir enerji kaynakları kullanımının ekonomik büyümeyi artırdığı görülmektedir. Bu bulgu fosil yakıtların ikamesi olan yenilenebilir enerji kaynakları kullanımının teşvik edilmesi açısından önem taşımaktadır. Böylece bir yandan ekonomik büyüme sağlanırken diğer yandan çevresel kirliliğin önüne geçilmiş olacaktır. Sürdürülebilir bir çevrenin devamlılığı için söz konusu temiz enerji kaynaklarının kullanımı olmazsa olmaz unsurlar arasında yer almaktadır. Benzer şekilde fosil yakıtlarında ekonomik büyümeye olumlu katkısının olduğu saptanmıştır. Gerek temiz enerji kaynaklarının kullanımı gerekse kirletici kaynakların kullanımı ekonomik büyüme üzerinde olumlu etkilere sahiptir. Bu bağlamda her iki kaynak türünün de ekonomik büyümenin itici güçleri arasında yer aldığı söylenilebilir. Ancak fosil yakıtların ekonomik büyümeyi sağlamasının yanı sıra çevre üzerine olan zararları göz önüne alındığında, mutlak suretle söz konusu büyümenin temiz enerji kaynakları ile gerçekleştirilmesi gerekliliği ortaya çıkmaktadır. Ayrıca beklenildiği üzere emek, sermaye ve nüfus artışının da ekonomik büyümeyi artırdığı sonucuna ulaşılmıştır. Tüm bu bulgular değerlendirildiğinde SDG 7 ve SDG 8 hedefleri arasında uyumsuzluk olmadığı görülmektedir. Ekonomik büyümenin itici güçleri arasında yer alan fosil yakıtların yenilenebilir enerjiler ile ikame edilebilmesinden ötürü hem temiz bir çevre hem de ekonomik büyümenin sağlanması adına yenilenebilir enerji kaynaklarının kullanımının yaygınlaştırılması önerilmektedir. Bunun için yenilenebilir enerji teknolojilerinin ve yatırımlarının geliştirilmesine yönelik politikalar ve diğer mali yardımlar yapılabilir. Aynı zamanda sübvansiyonlar ve vergi indirimleri gibi desteklerle yenilenebilir enerji kaynaklarının kullanımı teşvik edilmelidir. Yenilenebilir enerji kaynaklarının yaygınlaştırılması sadece ekonomik büyümeyi sağlamakla kalmayarak aynı zamanda çevresel bozulmanın da önüne geçecektir. Ayrıca özellikle yeni sanayileşen ülkelere gelişmiş ülkelerle rekabet edebilme avantajı sağlayabilecektir. Elif Koçak \| 95 Elif Koçak \| 95 95 Kaynakça Apergis, N., & Payne, J. E. (2012). 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Economic welfare impacts from renewable energy con- sumption: The China experience. Renewable and sustainable energy Re- views, 15(9), 5120-5128. Farhani, S. (2013). Renewable energy consumption, economic growth and CO2 emissions: Evidence from selected MENA countries. Energy Econo- mics Letters, 1(2), 24-41. Grossman, G. M.,& Krueger, A. B. (1991). Environmental impacts of a North American free trade agreement. Harris, J. M. (2000). Basic principles of sustainable development. Dimensions of Sustainable Developmnet, 21-41. Inglesi-Lotz, R. (2016). The impact of renewable energy consumption to eco- nomic growth: A panel data application. Energy economics, 53, 58-63. Jebli, M. B., & Youssef, S. B. (2015). Output, renewable and non-renewable energy consumption and international trade: Evidence from a panel of 69 countries. Renewable energy, 83, 799-808. Jiang, J., Zhu, S., Gao, S., Aslam, B., & Wang, W. (2023). 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International Labour Organization. Rafindadi, A. A., & Ozturk, I. (2017). 1 Prof. Dr. Dicle Üniversitesi, İktisadi ve İdari Bilimler Fakültesi, İktisat Bölümü, kayamehmet@dicle.edu.tr, ORCID: 0000-0002-9495-4141 2 Dr. Dicle Üniversitesi, muhammed.celik@dicle.edu.tr, ORCİD: 0000-0003-3189-0141 Özet Türkiye’de sanayi sektörünün gelişimini anlamak için dönemsel olarak ele almak gerekir. Bu nedenle bu çalışmada da Türkiye’de sanayileşme; Cumhuriyet dönemi öncesi, Atatürk dönemi ve sonrasında yaşanan gelişmeler, planlı dönem ve piyasa ekonomisine geçiş dönemi çerçevesinde incelenmiştir. Türkiye’ de sanayi sektörü adeta sıfırdan başlayıp, şu an istihdamı en çok etkileyen ikinci sektör konumundadır. Bunun yanında, ihracatın %75’ ine yakınını gerçekleştiren ve milli gelirin nerde ise dörtte birinde pay sahibidir. Bu minvalde; hem ezeli ve ebedi kalkınma hedefini gerçekleştirmek hem de stratejik konumu, birçok sektörü etkilemesi, milli gelir ve istihdam üzerindeki payı itibariyle sanayi sektörü, Türkiye için kesinlikle ihmal kabul etmez bir sektördür. Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Mehmet Kaya1 Muhammed Çelik2 Mehmet Kaya1 Muhammed Çelik2 5.Sonuç Impacts of renewable energy consump- tion on the German economic growth: Evidence from combined coin- tegration test. Renewable and Sustainable Energy Reviews, 75, 1130-1141. Salim, R. A., Hassan, K., & Shafiei, S. (2014). Renewable and non-renewab- le energy consumption and economic activities: Further evidence from OECD countries. Energy economics, 44, 350-360. Sari, R., & Soytas, U. (2004). Disaggregate energy consumption, employment and income in Turkey. Energy economics, 26(3), 335-344. Shafik, N.,& Bandyopadhyay, S. (1992). Economic growth and environmen- tal quality: time-series and cross-country evidence (Vol. 904). World Bank Publications. Shahbaz, M., Loganathan, N., Zeshan, M., & Zaman, K. (2015). Does re- newable energy consumption add in economic growth? An application of auto-regressive distributed lag model in Pakistan. Renewable and Susta- inable Energy Reviews, 44, 576-585. Solomon, C. G., Salas, R. N., Malina, D., Sacks, C. A., Hardin, C. C., Prewitt, E., & Rubin, E. J. (2022). Fossil-fuel pollution and climate change—a new NEJM group series. New England Journal of Medicine, 386(24), 2328-2329. Zou, C., Zhao, Q., Zhang, G., & Xiong, B. (2016). Energy revolution: From a fossil energy era to a new energy era. Natural Gas Industry B, 3(1), 1-11. Bölüm 7 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 1.Giriş Ekonomiler tarih boyunca önemli dönüşümlerden geçmişler ve bu dönüşümlerle kalıcı gelir artışı yaratmışlardır. Örneğin MÖ 10.000’lerde olan tarım devrimiyle yerleşik hayata geçildiğinde kalıcı bir ekonomik değer artışı yaşanmıştır. Hakeza 18. yüzyılın sonlarına doğru önce İngiltere ve Avrupa’da, sonra da Amerika’da gerçekleşen sanayi devrimi de ekonomik ve toplumsal yapıyı kalıcı olarak değiştirmiştir. Birinci Sanayi Devrimi olarak adlandırılan bu gelişmeyle, emek yoğun üretimden sermaye yoğun üretime geçilmiş ve üretim de evlerden ve küçük atölyelerden fabrikalara taşınmıştır. Bu sayede hammaddelerin işlenerek ürün haline getirilmesi sürecini kapsayan sanayi sektörü de üretim alanları arasında yer almıştır. Sanayileşme; belirlenen bir alanda sanayi sektörü ile ilgili hareketliliği öne https://doi.org/10.58830/ozgur.pub401.c1755 97 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 98 çıkarma amacıyla yürütülen sosyal, ekonomik ve mühendisliğe ilişkin iş ve işlemlerin tamamı olarak tanımlanmaktadır. Nitekim sanayi sektörünün geliştiği ülkelerde tarım sektöründe çalışan nüfus imalat sanayine geçmiş, bu sektörde artan üretkenlik de ekonomik büyümeyi tetiklemiştir. Ekonomik büyümeyle birlikte hizmet sektörü de gelişir ve toplam üretimdeki payı artar; ancak teknolojik ilerleme ve yenilikçilik daha çok sanayi sektöründe ortaya çıktığı için sanayi, büyümenin ve gelişmenin motoru olmaya devam eder. Ekonomik olarak ilerlemiş ülkeler aynı zamanda sanayileşmede ilerlemiş ülke olmaları, sanayileşme ile ekonomik gelişim arasındaki ilişkiyi de ortaya koymaktadır. Sanayi sektörünün bu özelliği dolayısıyla “sanayileşmiş ülke” ve “gelişmiş ülke” kavramları birbirinin yerine kullanılır. Aynı şekilde gelişmekte olan ülkeler de genellikle sanayileşme sürecini henüz tamamlamamış ülkeler olarak kabul edilir (Yağmur, 2020:61-61, Koç ve diğerleri, 2018:1). Osmanlı İmparatorluğu’nda da 1800’li yıllardan itibaren Avrupa’da başlayan sanayileşmeye yönelik önemli adımlar atılmıştır. Bu bağlamda bazı sanayi yatırımları yapılmış olsa da, kısmen uygulama hatalarından kısmen de dönemin politik koşullarının etkisiyle Osmanlı sanayileşmiş bir ülke olamamıştır. Zaten Osmanlı’da, imparatorluk toprağını ve sınırlarını korumaktan ve basit el sanatları ve tarımsal faaliyetlerle ekonomisini döndürmeye çalışmaktan başka bir şey öngörmeyen bir devlet yönetiminin aklına, her ne kadar gelişmeleri yakından takip etse de, sahip oldukları iktisadi zihniyetten dolayı İngilizlerin başardığı bir sanayileşmeyi başlaması mümkün görünmüyordu. Oysa Cumhuriyeti kuran kurucu akıl muasır medeniyetler seviyesine çıkmayı ilk hedef olarak ortaya koymuş, bunu gerçekleştirmek için ise gelişmiş ülkelerin izinden gitmek veya deneyimlerinde yararlanmak üzere onların ileri rejimini, iktisat politikalarını ve sanayileşme stratejilerini uygulaması gerekiyordu (Kalaycı vd.,2019:310). Bu itibar ile Türkiye kuruluşunun ilk yıllarında sanayileşmeye engel teşkil eden uygulamaları ortadan kaldırmaya ve anlaşma ile yapılan uygulamalarında sanayileşme üzerindeki olumsuz etkisini azaltmak amacıyla çeşitli kararlar yürürlüğe konulmuştur. Özellikle Cumhuriyetin ilk yıllarındaki Birinci sanayi planının önemli stratejik tercihi kalkınmanın sanayileşme ile sağlanabileceğini kabul etmesi ve bu doğrulduda uygulamalara geçilmesidir (Han, 1977). 2. Cumhuriyet Dönemi Öncesi Sanayi Sektörü Sanayi sektörü yıllar içerisinde; tecrübelerle, bilgi birikimi ve gelenek ile yerleşip gelişebilen bir sektördür. Bu nedenle Cumhuriyet dönemindeki sanayi sektörü incelenirken Cumhuriyet öncesi Osmanlı’nın son döneminde sanayi sektörünün durumunu gelişimini ortaya koymak gerekir. Mehmet Kaya / Muhammed Çelik \| 99 99 Mehmet Kaya / Muhammed Çelik \| 99 Osmanlı sanayisi özellikle ordu ve devlet kurumlarının ihtiyaçlarını karşılamak üzere faaliyetleri 16. ve 17. yüzyıllara dayandırmaktadır. Bu dönemde Osmanlı sanayisi; devlet sermayesi ile kurulan askeri fabrikalar, yerli özel kişilerin kurduğu işletmeler ve tekelci görünümdeki yabancı yatırımcılar olmak üzere üç bölüme ayırmak mümkündür. Bu dönemdeki işletmeler Avrupa ülkeleriyle mücadele edebilecek ölçekteydi. Ancak özellikle 18. yüzyılın sonlarına doğru Avrupa’ da yaşanmaya başlanan sanayi devrimi ile birlikte Osmanlı sanayisini rekabet gücünü zayıflatmıştı. Daha doğrusu 18. yüzyılın sonlarına doğru başlayıp 19. yüzyılın başların sıçramaya geçen Avrupa sanayisi Osmanlı sanayisin karşısında oldukça güçlü duruma geçmiş. Osmanlı sanayisi bu dönemde rekabet gücünü tamamen kaybetmiştir. Avrupa’da yaşanan bu sanayi devrimi makinalı üretime geçerken, Osmanlı sanayisinin emek piyasasına dayalı rekabet gücünü adeta yerle bir etmişti (İlkin, 1970). Bunun yanında Osmanlı sanayisi sanayi devrimine ayak uydurmak bir yana, bu süreçte özellikle İngiltere ile Balta Limanı Ticaret Anlaşması ve diğer Avrupa ülkeleri ile kapitülasyon tarzı gümrük anlaşmaları imzalaması, sanayi alanındaki çöküşü daha da hızlandırmıştır (Keleş, 2021). Sonraki dönemlerde sanayinin gelişmesi için “Islahı Sanayi Komisyonu (1860)” kurulması ve “Teşvik-i Sanayi Kanun-ı Muvakkatı (1913)” çıkarılması ile sanayinin teşvik edilmesine yönelik yapılan çalışmalar ise, kapitülasyonlar dolayısıyla başarısız olmuştur. Tablo 1. Cumhuriyet Öncesi Osmanlı Sanayi Sektörleri ve Tesisi Verileri SANAYİ SEKTÖRÜ TESİS SAYISI Gıda 78 Toprak 21 Deri 13 Ağaç 24 Dokuma 78 Kırtasiye 55 Kimya 13 Toplam 282 Kaynak: (Osmanlı Sanayi 1913-1915 Yılları Sanayi İstatistikleri 1978 sf 13-23 Aktaran: İlkin,1970:340). Tablo 1. Cumhuriyet Öncesi Osmanlı Sanayi Sektörleri ve Tesisi Verileri SANAYİ SEKTÖRÜ TESİS SAYISI Gıda 78 Toprak 21 Deri 13 Ağaç 24 Dokuma 78 Kırtasiye 55 Kimya 13 Toplam 282 Kaynak: (Osmanlı Sanayi 1913-1915 Yılları Sanayi İstatistikleri 1978 sf 13-23 Aktaran: İlkin,1970:340). Tablo 1. Cumhuriyet Öncesi Osmanlı Sanayi Sektörleri ve Tesisi Verileri Kaynak: (Osmanlı Sanayi 1913-1915 Yılları Sanayi İstatistikleri 1978 sf 13-23 Aktaran: İlkin,1970:340). Kaynak: (Osmanlı Sanayi 1913-1915 Yılları Sanayi İstatistikleri 1978 sf 13-23 Aktaran: İlkin,1970:340). Osmanlı dönemi ekonomik hayatında tarımın ezici bir ağırlığı vardı. Osmanlı diğer sektörlere biraz yabancı idi. Tablo 1’ den de görüldüğü üzere toplam 282 sanayi tesisi bulunmaktaydı. Her ne kadar bunlara sanayi Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 100 tesisi denilse de aslında büyük çoğunluğu da imalathane şeklinde üretim yapan tesislerdir. 2. Cumhuriyet Dönemi Öncesi Sanayi Sektörü Bu tesislerdeki üretimin üç ayrı şekilde örgütlendiğini söyleyebiliriz. kırsal alanlardaki tarım dışı üretim faaliyetleri ve şehirlerde imalathaneler etrafında biriken zanaatler, basit el aletlerine dayanan geleneksel teknolojileri kullanıyordu. Üretimin üçüncü örgütlenme şekli ise, Sanayi Devrimi sonrasında Avrupa’dan ithal edilen makineleri kullanan ve ücretli işçi kiralayan imalathaneler ya da fabrikalardır. Bu fabrikalar da esas olarak İstanbul ve bir ölçüde de İzmir ile Adana yörelerinde faaliyet gösteriyordu. Birinci Dünya Savaşı öncesinde bu üç yöredeki büyük ölçekli sanayi işletmelerinde anacak beş bin, bu tesislerin tamamında ise toplamda 14000 kişi istihdam edilmekteydi (Alpağut, 2022:248, Pamuk 2012:19-19). Kuşkusuz Osmanlının nüfusu düşünüldüğünde, sanayi sektöründeki istihdam düşüklüğü bu sektörün geri kalmışlığını da gözler önüne sermekteydi. Osmanlı’nın Sanayi Devrimi’nin dışında kalmasında, yukarıda da belirtildiği gibi, devlet anlayışı da etkili olmuştur. Nitekim Avrupa’da sanayi atılımını doğuran ve destekleyen unsurlar incelendiğinde bu durum daha da netleşmektedir. Sanayileşmenin ön koşullarını ortaya koyan bu unsurların başlıcaları şöyle özetenebilir. i- Siyasi Altayapı: Sürekli bir istikrar ortamı ve yeterli serbest piyasa koşulları olmadan, özellikle mülkiyete ve sermayeye hukuki güvenceler sağlanmadan, büyük ve uzun ömürlü yatırımları gerektiren sanayileşmenin gerçkleşmesi beklenemz. ii-Fiziki Altyapı ve Diğer Unsurlari: Sanayileşmenin başlayıp yayılabilmesi, büyük ve genişleyen pazarlar gerektirir, bu da yeterli ulaşım, iletişim ve finans olanakları ister. Bunlardan daha önemlisi ise, nüfusun asagari bir eğitim altyapısına, teknik eğitim düzeyine sahip olmasıdır (Yenal, 2010:30). 3. Atatürk Dönemi: İktisat Kongresi ve Sanayi Planları Cumhuriyet döneminin ilk yılları Osmanlı devletinden devralınan siyasi, askeri ve ekonomik sorunların çözümüne odaklanılmıştır. Bu dönemde çoğunlukla Osmanlı ekonomisini esir alan uygulamalar ve bu uygulamaların sebep olduğu sıkıntıların bertaraf edilmesine çalışılmıştır. Askeri ve siyasi zaferle kurulan yeni Cumhuriyetin tam bağımsızlığı ancak ekonomik gelişmenin ve ilerlemenin sağlanmasıyla sağlanabilirdi. Bu nedenle öncelikle Osmanlı ekonomisinin gelişimini olumsuz etkileyen uygulama ve anlaşmalara bu dönemde son verilmiştir. Özellikle 1923-1930 yılları arasında alınan kararlar ve uygulamalar tam ekonomik bağımsızlık ve süratli bir ekonomik kalkınma iki temel iktisadi hedef belirlendiği söylenebilir (Durmuş ve Aydemir, 2016:16). Cumhuriyetin kuruluşundan bağlayarak geçen ilk on yılda uygulanan genel ekonomi politikalarının çerçevesini ise, İzmir İktisat 101 Mehmet Kaya / Muhammed Çelik \| Kongresi’nde alınan kararlar ve Lozan’da belirlenen sınırlamalar çizmiştir (Kepenek ve Yentürk, 2003:35). Atatürk henüz Cumhuriyeti ilan etmeden, yeni Türkiye’nin izleyeceği iktisat rejimi ve sanayileşme politikası ile ilgili yol haritasını belirlemek için sektör temsilcilerinden oluşan bir iktisat kongresi düzenlemiştir (Kalaycı vd., 2019:). Nitekim Kılıçbay’ın (1991:62) da belirttiği gibi,kongre zamanın şartlarına ve Türkiye ekonomisinin gerçeklerine uyan bir model çalışması veya model denemesidir. Model ise sade ve gerçekçi olup, doktrinci bir karaktere sahip olmaktan çok iktisadi gerçeklerin, iktisat mantığının ve akılcılığının ürünüdür. Bu minvalde temel yaklaşımı ise ekonomide sanayileşmeye hız ve güç vermektir. Sanayileşmenin hedeflenmesindeki motifler ise şöyle sıralanabilir: - Tarım ürünleri dışında hemen her malı ithal etmek zorunda olan bir ülke durumundan kurtulmak, - Dünyanın gelişmiş ülkelerinin, sanayileşmiş ülkeler olduğu gözlemine dayanan ampirik bir yaklaşımla sanayileşmiş ülke ilerlemiş ülkedir tezini benimsemek, - Sanayileşme ile kültür ve medeniyet düzeyinin yüksekliği arasında yakın bir ilişkinin varlığını kabul etmek, - Nihayet, sanayileşmeyi “romantik bir ideal”, milli bir övünme kaynağı olarak kabul etmektir. Sanayileşme hedefini gerçekleştirmek üzere kongredeki sanayi grubunda, sanayinin teşvik edilmesi, korunması ve desteklenmesine yönelik bir dizi kararlar alınmıştır. Bunlar: (Evsile, 2018:110): - Hammaddesi yurtiçinde yetiştirilen veya yetiştirilebilen sanayi dalları kurulmalıdır. - Hammaddesi yurtiçinde yetiştirilen veya yetiştirilebilen sanayi dalları kurulmalıdır. - El işçiliğinden ve küçük imalattan süratle fabrikaya ve büyük işletmelere geçilmelidir. - Devlet iktisadi görevleri bulunan bir organ haline gelmelidir. - Özel sektör tarafından kurulamayan teşebbüsler devletçe ele alınmalıdır. - Özel teşebbüse finans desteği sağlayacak bir devlet bankası kurulmalıdır. - Dış rekabete dayanabilmek ve mücadele edebilmek için sanayinin toplu ve bir bütün olarak kurulması gerekmektedir. İzmir İktisat kongresinde sanayi gurubunda alınan kararlar itibariyle yerli sanayiyi geliştirmek amacıyla stratejik sanayi yatırımlarının önemi ve Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 102 gerekliliği vurgulandığı görülmektedir. 3. Atatürk Dönemi: İktisat Kongresi ve Sanayi Planları Alınan kararlar ve tartışılan konular itibariyle İzmir İktisat Kongresi yeni Cumhuriyet için kalkınma modeli arayışını da göstermektedir. Aynı zamanda bu kongrede alınan kararlar; sanayinin teşviki, korunması ve finansmanı olmak üzere üç ana başlıkta toplandığı görülmektedir (TMMOB Makine Mühendisleri Odası, 2009:9). Her ne kadar İzmir İktisat Kongresinde alınan tüm kararlar hükümet için bağlayıcı olmasa da, özellikle sanayi grubunda alınan tavsiye kararlarının hükümet programına büyük ölçüde yön verdiği görülmektedir. Nitekim Cumhuriyet Halk Fırkası tarafından İş Bankasının kurulması, aşar vergisinin kaldırılması, kazanç vergisinin kabulü, Teşviki Sanayi Kanunu, Gümrük Kanunu, Menkul Kıymetler Borsası Kanunun çıkarılması bu kongrede alınan kararların hükümet programına yön verdiği görülmektedir (Koç, 2000). Bu dönemde (1927) yapılan sanayi tesisi sayımında; %43,6 gıda ve tütün, %23,8 dokuma sanayisi, %22,6 maden, metal, toprak ve makine sanayisi yan dallarında ve yaklaşık %10’ nu da ağaç sanayisinde faaliyet göstemekteydi. Sanayi sektöründe üretilen katma değer 201 milyon TL idi (Şahin, 2006: 43). Tablo 2. Atatürk Dönemi Sanayi Sektörü’ nün GSYİH İçerisindeki Payı YILLAR SANAYİ SEKTÖRÜ(Yüzde) YAPIM SANAYİ(Yüzde) 1923 13,2 12,3 1930 11,4 10,3 1935 18,1 16,4 1940 18,9 17,1 Kaynak: (Han, 1977:107) Tablo 2. Atatürk Dönemi Sanayi Sektörü’ nün GSYİH İçerisindeki Payı Kaynak: (Han, 1977:107) Bu dönem yeni kurulan bir sanayi sektörü için iyi bir hükümet programı hazırlandığı söylenebilir. Bu program küresel ekonomiye yeni cumhuriyetin ekonomi politikasını, ekonomiye yaklaşım tarzını belirlemesi ile yasal ve fiziki altyapıyı hazırlaması açısından oldukça önemlidir. Aynı zamanda günün koşulları düşünüldüğünde kapsamlı bir plan ve programdır. Ancak bu dönemde yaşanan ve küresel etkileri çok yıkıcı olan 1929 küresel ekonomik kriz, bu planın tam olarak uygulanmasını zorlaştırmış ve sanayi sektöründe istenen düzeyin yakalanmasını engellemiştir. Dünya genelinde uygulanan liberal kapitalist sistemin en önemli sınavı olan büyük buhranın küresel ekonomiler üzerindeki olumsuz ve yıkıcı etkisi hükümetleri liberal politikalardan, devletçi politikalara 103 Mehmet Kaya / Muhammed Çelik \| 103 yönlendirmiştir. Nitekim Türkiye Cumhuriyetinin hükümet çevrelerinde 1923-1929 dönemindeki sanayileşmenin yetersiz kaldığı, bu nedenle ana tüketim mallarında bir ithal ikameci bir politika benimsenerek sanayileşme sürecinin hızlandırılması gerektiği konusunda fikir birliği oluşmuştur (Yücel, 2014:21). Bu planın hazırlıkları 1929 yılında kurulan Ali İktisat Meclisinin kurulmasıyla başlamıştır. Bu dönemde devletçi politikalara geçiş fikrinin ağır bastığı ve planların buna göre hazırlanmaya başlandığı bir dönemdir (Akyıldız ve Eroğlu, 2004:48). 3. Atatürk Dönemi: İktisat Kongresi ve Sanayi Planları Bu dönemde devletçi politikalara geçişin bazı sebepleri bulunmaktaydı bunları şu şekilde sıralayabiliriz (Özçelik ve Tuncer, 2007; Durmuş ve Aydemir, 2016); -Küresel ekonomilerle uyumlu olarak uygulanan liberal politikaların başarısız olduğu düşüncesinin yaygınlığı -Büyük Buhranın olumsuz etkileri, -Liberal politikaların 1929 ekonomik krizine çözüm üretememesi nedeniyle devletin müdahale etmesi gerektiği fikrinin yaygınlaşması, -Liberal politikaların 1929 ekonomik krizine çözüm üretememesi nedeniyle devletin müdahale etmesi gerektiği fikrinin yaygınlaşması, -SSCB’ de uygulanan devletçi ve kapalı politikalrın başarılı olduğu düşüncesinin yaygınlaşması, -SSCB’ de uygulanan devletçi ve kapalı politikalrın başarılı olduğu düşüncesinin yaygınlaşması, Bu düşüncelerin yaygınlığı uygulanan politikaların değiştirilmesi gerektiği fikrinin ağır basmasına neden olmaktaydı. Bu doğrultuda Birinci Beş Yıllık Sanayi Planı (BBYSP) hazırlanmış ve politikalar buna göre belirlenmiştir. Temel hammaddesi ülkede yetişen ve kısa sürede üretilebilen ürünlerin sanayisi ele alınmış ve bu sanayilerin büyük ölçekli olması nedeniyle devlet tarafından kurulması gerektiği benimsenmiştir (İnan, 1972:16). BBYSP amaçlarının temel niteliğine göre salt bir sanayi planı olup, tümüyle ekonomiye da amaçlar taşımadığından bir makro plan niteliği göstermez. Buna karşın Planda ele alınan kuruluşların işgücü ve diğer girdi gereksinimleriyle ilgili öngörülere de yer verilmiştir. Ayrıca Plan ile kurulması tasarlanan sanayi; dokuma, maden işlem, kağıt, kimya ve taş-toprak olmak beş grupta toplanmıştır (Kepenek ve Yentürk, 2003:68). Sanayi planı çok hızlı bir şekilde uygulamaya konulmuş, iki yılda ondan fazla fabrikanın temelleri atılımış, bu fabrikalara nitelikli eleman yerleştirilmiş ve fabrikalar iki-üç yıl gibi kısa bir sürede üretime geçmiştir. Kuşkusuz insan kaynaklarının kıt ve kurumların zayıf olduğu koşullarda bu, büyük bir başarıydı. Orta ve küçük ölçekli işletmeler de hesaba katıldığında,1930’ların devletçilik politikası ile sanayileşmenin anlamı daha da dikkate değer bir hal almaktadır (Yenal, 2010:86). Nitekim plan çerçevesinde kurulan sanayiler sayesinde ülke üç beyazda (dokuma, un ve şekerde) dışa bağımlılıktan 104 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme kurtuldu. Sanayinin milli gelirdeki payı 1929’da % 10 iken 1939’da % 18’e çıktı (Somel, 2004:8). kurtuldu. Sanayinin milli gelirdeki payı 1929’da % 10 iken 1939’da % 18’e çıktı (Somel, 2004:8). BBYSB’nin uygulanmasıından elde edilen başarının etkisiyle,1936 yılının ardından İkinci Beş Yıllık Sanayileşem Planı (İBYSB) için hazırlıklar yapılmaya başlanmıştır. İBYSP,ilk plandan farklı olarak ara ve yatırım malları üretimine öncelik vermiş, ayrıca enerji, madencilik ve limanlar gibi altyapısal gelişmeyi de dikkate almıştır. Bu amaçla İBYSB gıda, kimya, makine ve deniz ulaşımı vb. alanlarda yaklaşık 112 milyon TL’lik yatırımı öngörmüştür. İBYSB bu nitelikleri ile aslında bir bakıma “kendine yeterlik” ilkesine önem verdiği ve birinci planının doğal bir uzantısı olduğu söylenebilir. Ancak İkinci Dünya Savaşı nedeniyle plan askıya alınmıştır (Kepenek ve Yentürk, 2003:69). 3. Atatürk Dönemi: İktisat Kongresi ve Sanayi Planları Kaldı ki Türkiye savaşa katılmasa da, sınırına kadar ulaşan savaşın ağır ekonomik etkilerine maruz kaldı. Erkeklerin silah altına alınması ve hammade, ara ve yatırım malı ithalatının azalması dolayısıyla, tarımsal üretim, örneğin buğday üretimi % 50, düştüğü gibi sanayi üretimi de, savaş ve onu izleyen üç yılda (1939-1948) yılda ortalama -% 0,9 ile tamamen duraklamıştır (Somel, 2004:9 ve Kazgan, 1999:89). 4. Planlı İthal İkameci Sanayileşme (1961-1979) 1930’lı yıllarda, zorunluluktan da olsa, uygulanan devletçilikle sanayileşme girişimi sonrdan çok eleştiri konusu da olmuştur. Özellikle fabrikların üretecekleri mal cinsi, yeri ve teknoloji seçimleri konusundaki tercihleri eleştirenler olduğu gibi, sanayi kesimine öncelik verilmesini, devletin doğrudan sanayi yatırımcılığına ve işletmeciliğine girmesini doğru bulmayanlar da olmuştur (Yenal, 2010:87). Bu kesimlerin eleştirilerinin sonuçları, 1946-1953 yılları arasında Cumhuriyet Halk Partisi (CHP) ve özellikle Demekrat Parti (DP) iktidarları nazarında dış faktörlerin de etkisiyle adeta vücut bulmuştur. Nitekim Türkiye İkinci Dünya ardından iki bloklu dünya düzeninde, ABD’nin başını çektiği batı/kapitalist blokunu tercih etmiş böylece 1930’dan beri uyguladığı devletçilikten zamanla uzaklaş(tırıl)mıştır. Çünkü Kepenek ve Yöntürk’ün de (2003:93) belirttiği gibi, ABD ve uluslararası kurumlar, savaş sonrası Avrupa’nın toparlanması için Truman Dokktrini programı çerçevesinde verilen Marshall desteklerinden yararlanmayı, ekonomi politiklarında kamu girişimciliğinin daraltılması özel sektörün ise gelişiminin desteklenmesi şartına bağlamışlardır. Ayrıca Türkiye’nin uluslararası işbölümünde, ağır sanayiden (demir-çelik, ağır kimya vb.) ziyade nispeten daha güçlü olduğu tarıma ve kısmen de hafif metal,inşaat malzemeleri, deri vb. tarzda sanayileşmeye yönelmeliydi. 105 Mehmet Kaya / Muhammed Çelik \| 1 Mehmet Kaya / Muhammed Çelik \| 105 Bu bağlamda İkinci Dünya Savaşı’ndan sonra 1946 yılında birinci ve ikinci sanayi planlarının devamı mahiyetinde, özellikle Sümerbank ve Etibank’ın projeleri demetinden oluşan, İvedi Sanayi Planı’ından vazgeçilmiş ve yerine 1947 yılında Vaner Planı diye anılan Türkiye Kalkınma Planı devreye sokulmuştur. Özellikle bütçe yetersizliğinden dolayı doğrudan uygulanmasa da bu plan; özel kesime öncelik veriyor, kamu kesimini, ulaştırma başta olmak üzere altyapıyı sınırlıyor ve dış finansmana dayanıyor kısaca dönemin egemen ekonomi politikasını yansıtıyordu (Kepenek ve Yentürk, 2003:91- 92). Ancak tüm bu ekonomi politikaları liberal ekonomi modelin benimsendiğini gösterse de, Boratav’a (2015:109,111) göre “ Liberal” DP, ekonomik zorlamalar sonucunda, bir yandan kontrollü bir dış ticaret rejimine, öte yandan da, tüketim malı ithalatındaki daralmaları telafi etmeye çalışan ve önemli ölçüde devlet yatırımlarıyla gerçekleştirilen bir ithal ikamesi politikasına angaje olmuştur. Hatta 1946’dan itibaren, özellikle 1951 ve 1952’de, dış ticaret açığının artması hükümeti, 1953 ve 1954’te Milli Koruma Kanunları kapsamında ithalatı sınırlandırmaya aynı zamanda ithal ikameci yatırımları arttırmaya ve desteklemeye sevk etmiştir. Bu durum, Pamuk (2010:250) Türkiye’nin koşulları dikkate alındığında tarım- sanayi konusunda aslında yapılacak bir tercih olmadığını da siyasal iktidarlara göstermiştir. Zaten Türkiye’nin sanayileşmesini geri plana itip, karşılaştırmalı üstünlüğe göre tarıma öncelik vermesi tavsiyeleri de havada kaldı. Başlangıçta hükümet ve etkili çevrelerde kabul bulsa da, özellikle iki önemli olay bu önerileri etkisiz kıldı: i- Dünya Bankasının sürece müdahalesine hükümetin karşı çıkması dolayısıyla kredilerin kesilmesidir. 4. Planlı İthal İkameci Sanayileşme (1961-1979) ii- İkincisi ise Türkiye’nin büyüme aürecinde olayların sanayileşmeyi çok karlı hale getirmesiydi; 1954 ve izleyen yıllarda ithalatın krediler kesilince düşmeye başlaması, iç tasarruf hadlerini sanayi lehine değiştirdi.Dış konjonktürün tarım lehine değiştirdiği fiyatlar da bu eğilimi destekledi. Bunun yanı sıra bu dönemde, Türkiye Ticaret Odaları, Sanayi Odaları ve Türkiye Borsalar Birliği’nin hazırlattığı “Türkiye’de Sanayileşme ve Yatırım Meseleleleri” raporu da sanayinin tarımdan daha verimli olduğunu, dolayısıyla öncelikli olması gerektiğini gösteriyordu (Kazgan, 1999:101-102). Tüm bunların sonucunda sanayileşme, 1950’lerin ortalarından itibaren tekrar ekonominin motor gücü haline gelmişve toplam üretim içindeki ağırlığı artış eğilimine girmiştir (Tablo.3). Öyle ki 1954 ve sonrasındaki döviz bunalımı yılarında, özel sektör denetiminde ithal ikamesi, sermaye çevreleri için birikimin sürdürülebileceği tek alternatif oluşturmuştur. Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 106 1960 sonrasında ise ithal ikamesi, planlama ve diğer kurumsal ve yasal düzenlemelerle birilikte resmiyet kazanmıştır (Pamuk, 2010:250). Nitekim 1960 askeri darbesinden sonra 1961 yılında planlı kalkınmayı sağlamak ve yapılacak yatırımları bir plan dahilinde gerçekleştirilmesini sağlamak amacıyla Devlet Planlama Teşkilatı (DPT) kurulmuştur. Tablo 3. Sanayi Sektörü’ nün GSYİH İçerisindeki payı YILLAR SANAYİ SEKTÖRÜ İMALAT SANAYİ 1945 16,3 14,1 1948 14 12,5 1949 14,9 13,1 1950 14,6 12,7 1951 13,5 11,7 1952 13,3 11,4 1953 13,5 11,7 1954 15,5 13,5 1955 16 13,9 1956 16,7 14,6 1957 16 14 1958 17 15 1959 18,2 15,5 1960 17,3 14,6 1961 18 15,3 1962 18,1 15,6 Kaynak: (Han, 1977:107) Tablo 3. Sanayi Sektörü’ nün GSYİH İçerisindeki payı Kaynak: (Han, 1977:107) Bu dönemdeki planlama anlayışı, 1930’lardaki sanayi planlarından farklıydı. İktsadi ve topumsal gelişim stratejisini uzun vadeli şekillendiren ve tüm yönlerini kapsayacak bütüncül nitelikte olan planlar, kamu için emredici özel sektör için ise yol göstericidir (Soyak, 1999:100). Bu suretle karma ekonomik modelin yapısına uygun olarak kamu ile özel sektör arasında işbölümü sağlanmaya çalışılmıştır. Öyle ki büyük ölçekli ve yüksek harcama gerektiren, demir-çelik, petro-kimya vb. alanlarda yatırmlar KİT’ler, tekstil ve gıda sanayi ile otomobil, buzdolabı vb. dayanıklı yatırm malllarının ise özel sektör eliyle yapılması öngörülmüştür (Yücel, 2017:235). Bunun yanında ilk dört planın diğer ortak özelikleri; i-tüm alanlarda yeniden düzenleme (reform) sözlerine karşın mevcut üretim yapısın veri almaları, ii-ekonominin her yıl belli bir hızla büyümesini temel amaç Mehmet Kaya / Muhammed Çelik \| 107 107 edinmeleri ve iii-sanayileşemeye önem vermeleridir (Kepenek ve Yentürk, 2003:144). i-Birinci Beş Yıllık Kalkınma Planında Sanayi Sektörü (1963-1967) Bu plan daha çok montaj sanayisinin geliştirilmesi şeklinde bir sanayileşme modeli öngörmüştür. İthalat edilen parçaların yerli parçalar ile desteklenerek parçaların birleştirilmesi esasına dayanan bir sanayi sektörü planlaması yapılmıştır. Bu planda;makina imalat,gemi yapım (tersaneler), demiryolu ve karayolu taşıtları, plastik işleme ve lastik,elektrikli makine, cihaz ve donanım ve kimyasal ürünler sanayi sektörleri öncelikli sektörler olarak ön plana çıkmıştır (TMMOB Makine Mühendisleri Odası, 2009:11; DPB, 1963). ii-İkinci Beş Yıllık Kalkınma Planında Sanayi Sektörü (1968-1972) Bu planda bir önceki planda olduğu gibi sanayi sektöründe ithal ikameci sanayileşmeye devam etme kararı alınmıştır. Bu dönemde sanayici kökenli 12 kişi bir araya gelerek Türk Sanayicileri ve İşadamları Derneğini (TÜSİAD) kurmuştur (Uzunkaya, 2020:13). Birinci plandaki öncelikli sektörlere ilave olarak;tekstil ve konfeksiyon,deri eşya, orman ürünleri,topraktan mamul eşya sanayileri bu planda öncelikli sektörler arasında yer almıştır (TMMOB Makine Mühendisleri Odası, 2009:11). iii-Üçüncü Beş Yıllık Kalkınma Planında Sanayi Sektörü (1973- 1978) Bu dönem petrol krizi, finansman darboğazı ve Kıbrıs Barış Harekatı dönemini kapsaması açısından sanayi sektörünü ciddi manada etkileyebilecek bir karara rastlanılmamıştır. Ancak bu kalkınma planında ilk olarak bölgesel kalkınma ve kalkınmada öncelikli bölgeler kavramları ön plana çıkmıştır. iv-Dördüncü Beş Yıllık Kalkınma Planında Sanayi Sektörü (1979- 1983) Bu kalkınma planında gıda sanayisini destekleyici şekilde gıda üretimi, kooperatifleşmeyi ve gıda ihracatını artırıcı desteklerin sağlanması palnlanmıştır. Diğer sanayi dallarında da destekler ihracat şartlarına bağlanmıştır (DPT, 1979). 1980 öncesinin kalkınma planlarında; %7 (BKP,İKP) ya da % 7,2 ve 8,2 (ÜKP ve DKP) olarak öngrülen büyüme oranları temel belrileyici olmuş ve diğer amaçlar da buna göre düzenlenmiştir. Nitekim ilk üç plandaki büyüme oranlarına bağlı olarak, yatırımların milli gelir içindeki payları sırasıy; %18,3, %21,3 ve %21,9 olarak tasarlanmıştır. Söz konusu yatırımların yaklaşık % 90’ı da gerçekleşmiştir (Kepenek ve Yentürk, 2003:176-177). Ayrıca Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 108 ilk iki planda büyüme hedeflerine hemen hemen ulaşılırken, üçüncü plan için aynı şey söylenemez. Sanayide ise her üç planda da büyüme hedefinin altında kalınmışsa da, kısmi bir başarıdan söz etmek mümkündür. Zaten kriz dolayısıyla uygulanamyan dördüncü planın yerine konulan istkrar programının sonuçları ise, her anlamda hayal kırıkılığı yaratmıştır (Tablo 4.). Tablo 4. Plan Dönemlerinde Hedef-Sonuç Tablosu(Plan Döneminde Hedeflenen-(H), Gerçekleşen (G) ve Sektörel Büyüme (Yıllık Ortalama %) Sektör BBYKP(1963- İBYKP(1968- ÜBYKP(1973- 1978 1967 1972) 1977) Programı H G H G H G H G Tarım 4,2 3 4,2 2 3,7 1 4,1 3 Sanayi 12,3 10,9 12 9 11,2 9 8,8 3 Hizmetler 6,8 7,2 6,3 7 7,7 7 - 0 GSMH 7 6,7 7 6 8 5 6,1 1 Kaynak: T.C. Cumhurbaşkanlığı Strateji ve Bütçe Daire Başkanlığı, 2015 Tablo 4. Plan Dönemlerinde Hedef-Sonuç Tablosu(Plan Döneminde Hedeflenen-(H), Gerçekleşen (G) ve Sektörel Büyüme (Yıllık Ortalama %) Kaynak: T.C. Cumhurbaşkanlığı Strateji ve Bütçe Daire Başkanlığı, 2015 Dönem süresince tarımın GSMH’daki payı düşerken, sanayi ve hizmetler sektörünün payları artsa da veriler, değişimin hala istenilen düzeyde olmadığını göstermektedir. 3 Dışa açık büyüme ile ihracata dönük büyüme ayanı anlama geldiği halde ithal- ikamesiyle büyüme ve içe dönük büyüme ise eş anlamlı sayılmamalıdır ( Ayrıntılar için bkz. Kazgan,1 (Kazgan, Ekonomide Dışa Açık Büyüme, 1988). ii-İkinci Beş Yıllık Kalkınma Planında Sanayi Sektörü (1968-1972) Nitekim tarımın payı % 36’dan % 27’ye düşmüş iken, sanayinin %18’den % 20’ye ve hizmetlerin de % 46’dan % 54’e çıkmıştır (Toprak, 1999:71-72-73). Buna göre Türkiye ekonomisi, gelişmenin doğal akışı içinde, tarımdan sanayi ve hizmetler sektörüne doğru kaymıştır. Ancak 1980 yılı sonunda bile, tarımın GSMH’dan aldığı payın (% 24,2) sanayi sektörünün ( % 20,5) üzerinde olması, Türkiye ekonomisinin hala tarım ekonomisi vasfının devam ettiğini göstermektedir (Yıldırm, 2006). Son olarak bu dönemde diğer çevre ülkeler gibi, Türkiye’de, sanayileşmeye yönelik yatırımlarını borçlanarak finanse etti. Ancak 1977 yılı itibariyle borç stokunun cari işlemler gelirinin üstüne çıkması ve kısa vadeli borçların payının % 50’yi aşması ile başlayan borç krizi (Kazgan, 1999:127) ve onu izleyen enerji krizi ile ortaya çıkan stagflasyon sorunu, istikrarsız koalisyon hükümetleri ile de birleşince ekonomi ağır bir bunalıma girdi. Bu durumdan kurtuluşun çaresi 1980’den sonra, artık içerde değil dışarda (dışa açık) ve ithalatta değil ihracatta aranmıştır. 109 Mehmet Kaya / Muhammed Çelik \| 1 5.1980 Sonrası Dışa Açık/İhracat Dayalı Büyüme ve Sanayileşme3 Dışa açık ya da ihracata yönelik sanayileşme stratejileri, 1970’lerden sonra döviz darboğazına giren gelişmekte olan ülkelere sanayileşmelerinin dış pazara yönelik olması gerektiğini öneren bir stratejidir. Bu bağlamda Türkiye’de 1978-1979 yıllarında yaşadığı önemli döviz darboğazından sonra bu stratejiyi izlemiştir. Ancak bu politikalar aslında, 1980’lerde başlayan dönüşümün bir parçasıdır. Öyle ki ekonomik bunalımın yoğunlaşması ve 1977-1978 yıllarında alınan önlemler yetersiz kalması üzerine hükümet, 24 Ocak 1980’den itibaren yeni bir ekonomi politikasını uygulamaya başladı. Başlangıçta geçici bir dizi yeni önlemler ya da yaygın ifadesiyle “istikrar tedbirleri” olarak bilinen bu politika düzenlemeleri zamanla kalıcı bir nitelik kazandı ve uzun erimli bir iktisat politikasına dönüştü. Bu nedenle kamuoyunda 24 Ocak Kararları/Programı olarak bilinen bu politika (Kepenek ve Yentürk, 2003:363,196-197) aslında, 1970’li yıllarda IMF’nin dış tıkanma (döviz darboğazı) koşulları altında bunalan birçok azgelişmiş ülkeye dayattığı standart istikrar paketinden çok Dünya Bankası’nca geliştirilen tipik bir yapısal uyum programının bilinen tüm unsurlarını içermektedir. Kısaca Türkiye açısından 1923-1929 ve 1946-1953 yıllarında liberalizmle yaşanan geçici ilişki, artık liberalizmin 1980 modeli neo-liberalizmle kalıcı hale gelmiştir (Boratav, 2015:150-151). Bu bağlamda programın; i-ilk aşaması (1980-1983), KİT reformu, fiyat kontrolleri, kamusal mallarda sübvansiyonların kaldırılması vb., tedbirlerle, tipik bir istikrar paketi olarak uygulansa da, ii-ikinci aşaması (1984-1988) genelde ekonomiyi ve özelde mal ithalatının serbestleştirilmesi ve iii-üçüncü aşaması (1989-2001) ise TL’nin konvertibilitesi ve mali piyasaların serbestleşmesi, özelleştirme ve küreselleşmeyi kapsamıştır (Kazgan, 1999:146). Bu dönemin/zamanın ruhuna uygun olarak, 1983 yılında askeri rejimin uyguladığı kısıtlamalar altında yapılan seçimlerde, 24 Ocak programının mimarı Turgut Özal’ın kurduğu Anavatan Partisi (ANAP) tek başına iktidara geldi. Liberalleşme ve dışa açılma yönündeki politikalara ANAP döneminde de devam edildi (Pamuk, 2014:267) Bu sebeple “ANAP yılları” olarak nitelenen 1984-1988 kesiti, Özal iktidarının parlak dönemi diyebileceğimiz bir zaman dilimi (1984-1987) ve 1980’de benimsenen modelin tıkanışını ortaya koyan 1988 yılını içeriyor (Boratav, 2015:154). Yeni hükümet ekonomi programına uygun bir plan hazırlamak ve zaman kazanmak amacıyla, 1984 yılında yürürlüğe girmesi gereken Beşinci Beş Yıllık 110 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Kalkınma Planının (BBYKP) yerine, 1984 yılı için temel sektörlere yapılacak yatırım projelerini ve teşvikleri kapsayan geçiş programını uygulamaya koymuştur. Liberal uygulamaları esas alan hükümetlerce yürürlüğe konan beşinci ve sonraki kalkınma planlarında da öngörülen politikaların ortak hedefi ise piyasa ekonomisine geçişi hızlandırmak ve tamamlamak olarak belirlenmiştir (Yücel, 2017:316-317). Nitekim Türkiye ekonomisinin 1980-1983 dönüşümü ile başlayan dünya pazarlarına açılma süreci, 1989-1990’da tamamlanmıştır. Bu süreçte, öncelikle mal piyasaları dış pazarlara açılmış ve ticaret kotalarının koruması altındaki ithalat rejimi serbestleştirilmesidir. 5.1980 Sonrası Dışa Açık/İhracat Dayalı Büyüme ve Sanayileşme3 Döviz kuru yüksek bir devalüasyonun ardından esnekleştirilmiş ve dolaylı teşviklerle birleştirilerek sanayinin ihracata yönlendirilmesinde temel bir araç olmuştur. Kaldı ki. 1994’te Dünya Ticaret Antlaşması (1995’ten sonra Dünya Ticaret Örgütü) ve 1995’te de AB ile Gümrük Birliği (GB) Anlaşması’nın imzalanmasıyla ithalatın kontrol imkânlarından daha çok vazgeçilmiştir. Bu süreci ulusal mali piyasaların serbestleşmesi ve dış finans merkezlerine eklemlenmesiyle de Türkiye ekonomisi tamamen dışa açık bir ekonomi konumuna ulaşmıştır (Yeldan, 2003:25 ve Somel, 2004:12). İhracata dayalı sanayileşme stratejisi kapsamında uygulanan bu politikaların sonuçlarına ilişkin değerlendirmeleri, yine söz konusu programın, yukarıda bahsedilen uygulama sürecindeki aşamalarına referansla, aşağıdaki tablo aracılığıyla yapmak mümkündür. Mehmet Kaya / Muhammed Çelik \| 111 Tablo 5. GSMH ve Sektörel Büyüme Hızları ile GSMH’deki Sektörel Paylar Yıllar GSMH Tarım Sanayi Hizmetler Artış Oranı Hız Pay Hız Pay Hız Pay 1980 -2,8 1,3 23 -3,6 22 -3,6 55,4 1981 4,8 -1,8 22 9,9 23 5,9 55,9 1982 3,1 3,3 22 5,1 23 3,1 55,1 1983 4,2 -0,8 21 6,7 23 6,7 56,5 1984 7,1 0,6 20 10,5 23 8,2 57 1985 4,3 -0,3 19 6,5 23 5 58 1986 6,8 3,6 18 13,1 25 5,2 57 1987 9,8 0,4 17 9,2 24 13,2 59 1988 1,5 8 18 2,1 25 -0,8 57 1989 1,6 -7,7 16 4,9 26 3,2 58 1990 9,4 7 16 9,3 26 10,1 58 1991 0,3 0,6 16 2,9 27 0,5 57 1992 6,4 4,3 15 6,2 26 7,1 59 1993 8,1 -0,8 14 8,3 27 10,5 59 1994 -6,1 -0,6 15 -5,7 27 -7,6 58 1995 8 1,3 14 12,5 28 7,6 58 1996 7,1 4,6 14 6,8 28 7,9 58 1997 8,3 -2,2 13 10,2 28 9,9 59 1998 3,9 9,6 13 1,8 28 3,6 59 1999 -6,1 -5,6 13 -5,1 28 6,7 59 2000 6,3 3,8 13 6,2 28 7 59 Kaynak: (T.C. Cumhurbaşkanlığı Strateji ve Bütçe Başkanlığı,2015, TÜİK,2010 v Doğan, 2015:216) Kaynak: (T.C. Cumhurbaşkanlığı Strateji ve Bütçe Başkanlığı,2015, TÜİK,2010 ve Doğan, 2015:216) Kaynak: (T.C. Cumhurbaşkanlığı Strateji ve Bütçe Başkanlığı,2015, TÜİK,2010 ve Doğan, 2015:216) i-1980-1983 döneminde 1979 krizinin etkisiyle 1980’de ekonomide, sanayide ve hizmetlerde küçülme devam ederken tarımda çok cüzi bir artış yaşanmıştır. Dönemin sonunda, muhtemelen baz etkisiyle, ekonomi % 4,2 sanayi ise % 6,7 büyümüştür. Sanayinin payında az da olsa bir artış varken hizmet sektörü ağırlığını korumuştur. ii- Bu dönemde hem genel ekonomide hem de özellikle sanayide çift hanelere varan büyümeler ulaşmıştır. 4 Söz konusu dönemde (1980-2000) OECD ülkelerinde milli gelirde tarımın payı % 3, sanayini % 28 ve hizmetlerin ise %69 dolayında olmuştur (Toprak, vd.2001:245).. 4 Söz konusu dönemde (1980-2000) OECD ülkelerinde milli gelirde tarımın payı % 3, sanayini % hi l i i %6 d l d l T k d 5 ( ) g p y , y % 28 ve hizmetlerin ise %69 dolayında olmuştur (Toprak, vd.2001:245).. 5.1980 Sonrası Dışa Açık/İhracat Dayalı Büyüme ve Sanayileşme3 Özellikle kamuda altyapı ve sanayide özel sektör yatırımlarının teşviki ve imalat sanayindeki büyük atıl kapasitenin devreye girmesiyle sanayileşme hızı da göreli olarak hızlanmıştır. Sanayi sektörünün GSMH’daki payı pek değişmezken tarımın payı giderek azalmıştır. Buna karşılık bu dönemde de hizmet sektörü yüksek oranda büyümeye devam ederek % 60 paya erişmiştir (Kazgan, 1988:373 ve Şahin, 2010:202).Ancak 1980 reform sürecinin 1988’de ivmesini kaybettiğini Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 112 ekonominin de bir daralma içine girdiğini görmekteyiz. Bu nedenle 1990’da uygulamaya konan altıncı kalkınma planı izlenememiş ve hedeflerinin çok uzağında kalmıştır.. Özetle 1988’in verileri “ihracata yönelik büyüme” politikalarının sonuna erişildiğini göstermektedir (Yeldan, 2003:39). iii- Ekonomide 1988 yılında başlayan durgunluk 1989 yılında da devam etmiştir. Dönemin tamamında ekonomik büyüme istikrarsız bir seyir izlediği gibi ekonomi, artık neo-liberal düzenin tamamen hakimiyetine girmişken, 1994 ve 1999 yıllarında yaşanan krizlerle kesintiye uğramıştır. Söz konusu yıllar ekonomide her anlamda kayıp yılları olarak nitelemek mümkündür. Kaldı ki 1989-2000 yılları arasında tarımın GSMH’deki ağırlığı düşerken sanayinin ağırlığında ise bir artış olmamış, kararlı bir durağan eğilimi süregelmiştir. Dönem başında sanayi sektörünün toplam üretimdeki payı % 26 iken, dönem sonunda kısmi bir artışla % 28’ler düzeyinde karar kılmıştır4 (Toprak, Demir vd., 2001:245). Bu sonuçtan, ekonominin dışa açılma önceliklerinin reel üretim sektörlerinden çok finans ve kambiyo hizmetlerini kapsayacak politika değişikliklerin payı da olmuştur (Yeldan, 2003:39). Özellikle finansal küreselleşme sürecinde finansal piyasalarda yaşanan dalgalanmalar/krizler kuşkusuz reel ekonominin gelişimini olumsuz etkilemiştir. iii- Ekonomide 1988 yılında başlayan durgunluk 1989 yılında da devam etmiştir. Dönemin tamamında ekonomik büyüme istikrarsız bir seyir izlediği gibi ekonomi, artık neo-liberal düzenin tamamen hakimiyetine girmişken, 1994 ve 1999 yıllarında yaşanan krizlerle kesintiye uğramıştır. Söz konusu yıllar ekonomide her anlamda kayıp yılları olarak nitelemek mümkündür. 1980 sonrasındaki sanayileşmenin gelişimini aynı zamanda söz konusu dönemdeki ihracat tutarındaki ve ihracatın bileşimindeki değişimden de görmek mümkündür. Tablo 6. 5.1980 Sonrası Dışa Açık/İhracat Dayalı Büyüme ve Sanayileşme3 İhracatın Sektörel Dağılım (000000$) Sektörler 1983 1986 1989 1992 1995 1998 2000 Tarım (%) 1187(33,4) 1785(24,9) 2012 (18) 21348(14,8) 2133(10) 2670(10) 1973(7,2) Sanayi (%) 3666(66,6) 5393(72,1) 9171(82) 12286(85,2) 19089(90) 23873(60) 25340(92,8) Toplam 5503 7178 11183 14420 21222 26543 27313 Kaynak: (DİE, İstatistiki Göstergeler, 2003:416 ve DİE Ekonomik Göstergeler, 2005:139, aktaran Yıldırım,2006) Kaynak: (DİE, İstatistiki Göstergeler, 2003:416 ve DİE Ekonomik Göstergeler, 2005:139, aktaran Yıldırım,2006) Kaynak: (DİE, İstatistiki Göstergeler, 2003:416 ve DİE Ekonomik Göstergeler, 2005:139, aktaran Yıldırım,2006) Tablodan da anlaşılacağı üzere, 1980 öncesindekinden farklı olarak tarımsal malların ihracattaki payı azalırken, ekonomik gelişmenin doğal akışına bağlı olarak sanayinin üretimde olduğu gibi ihracattaki payı da 113 Mehmet Kaya / Muhammed Çelik \| 1 zamanla artmıştır. İhracatın da arması “ihracat dayalı büyüme” modelindeki amacın hasıl olduğunun sinyali olarak öngörülebilir. Bunun yanında, imalat sanayi yatırımlarının GSMH ve toplam yatırımlar içindeki payının seyrinde de sanayileşmenin gelişim çizgisine ilişkin ipuçları bulunmaktadır. Nitekim 1980’ler 1990’ların imalat sanayi yatırımları/GSMH oranı sırasıyla; % 20,2 ve 21,2 olarak; imalat yatırımları/toplam yatırım oranı ise;% 24,2 ve % 20,3 olarak gerçekleşmiştir. Birinci oran için cüzi bir artış söz konusu iken ikinci oranın düşmesi dikkat çekicidir ((Baştav, 2012:307). Tüm bunların sonunda, yaklaşık çeyrek asra yaklaşan ihracata dayalı modelinin sanayi sektöründe önemli bir yapısal dönüşümü gerçekleştirmediği ve özellikle 1990’larda itibaren seçici sanayi politikalarından vazgeçildiği, genel, uluslararası tamayüle uygun değişen kısa süreli hedeflere yönelen bir sanayi yapısından söz etmek mümkündür. Ayrıca sanayinin üretim yapısı emek ve kaynak ağırlı sektörlerde yoğunlaşmakta olup vasıflı işgücünün ve araştırmaya ayrılan kaynakların yetersizliği dolayısıyla ileri teknolojiye haliyle yüksek katma değerli üretim sürecine geçilememiştir. Sanayi sektörünün ithalata bağımlılığı, toplam ithalat içinde % 70’lere varan aramalı, ithalatı ve tasarrufların yetersizliğinden dolayı sağlıklı finansman sıkıntısı da dikkat çekicidir (Baştav, 2012:304). 6. 2001 Yılı ve Devamında Sanayileşme Politikaları Türkiye ekonomisi için 1990-2001 dönemi adeta bir krizler dönemi olarak nitelenebilir. Aslında 1980’le başlayan neoliberal/küreselleşme sürecinin yarattığı dışa açık ve finansallaşan/piyasalaşan ekonomi için, krizler son bulmayacağıya da artık sürpriz de olmayacağı söylenebilir (Kaya, 2020:43). Özellikle 1990’dan devralınan bozulmuş makroekonomik dengeler (yüksek enflasyon, borç stoku vb.) dolayısıyla oldukça kırılgan ekonomik yapı ve finansallaşmaya konnrolsüz ve denetimsiz bir şekilde yakalana bankacılık sektörü bu olasığı arttırmıştır. Ayrıca kısa süreli koalisyon hükümetleri dolayısyla yaşanan siyasi istikrarsızlık ve son tahlildeki siyasi krizin de etkisiyle, 2001 yılında Türkiye ekonomisi tarihin en büyük finansal krizini yaşamıştır. Ancak (Eğilmez,2018:151-152) krizden çıkışı oldukça hızlı oldu.ve bir yıl sonra yeniden büyümeye geri dönmeyi başardı.Bunun için 2001 ve 2002 yıllarında IMF’nin hazırlık ve uygulama surasında olduğu kadar finansmanında da destek olduğu istikrar programı ( Güçlü Ekonomiye Geçiş Programı-GEGP) uygulandı. 2002 yılında iktidara gelen Adalet ve Kalkınma Partisi (AKP) de bu programı uygulamaya devam etti. Sonraki yıllarda sağlanan makroekonomik ve finansal istikrar sayesinde yatırım ortamı iyileşti.Öyle ki net doğrudan 114 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme yatırım girişleri 2006’da GSYH’nın % 3,65 ile rekor düzeyde gerçekleşti. Ancak 2008 Küresel Finans Krizi ile sona eren küresel lkidiite bolluğu ile Türkiye’nin yakın coğrafyasında ve içeride yaşana siyasi gelişmelerin yanı sıra kaynak dağıtımı ve yönetimi konusundaki tercihlere, inşaat sektörüne başlayan yönelim gibi, bağlı olarak, sanayileşmeyi daha yukarı taşıyacak politikaların izlenmesinde yavaş kalındı (Yülek ve Yağmur, 2020:286). Bu sürece damgasını vuran neo-liberal politikların devletin ekonomideki gücünü törpülemesi ve tüm ekonomik kararlarında yol göstericiliğinin piyasanın yol göstericiliğinin benimsenmesi dolayısıyla kalkınma planlarınında bu minvalde, her alanda olduğu gibi,sanayide de özel sektörü teşvik etmek ya da uygun yatırım ortamı sağlamayı kapsadığnı belrtmek mümkündür. Nitekim bu husus planların ruhuna sinmiştir. ii-Dokuzuncu Beş Yıllık Kalkınma Planı ii-Dokuzuncu Beş Yıllık Kalkınma Planı Bu kalkınma planı 2007 yılında yürürlüğe girmiştir. Türkiyenin 2001- 2023 uzun süreli hedefleri perspektifiyle hazırlanan bir plandır. Bu bu plan sanayi sektöründe esas itibariyle yüksek katma değerli üretim yapısına geçişin sağlanması hedefiyle hazırlanmıştır. Böylece uluslar arası rekabet gücü artırılması hedeflenmiştir. Bölgesel gelişmişlik farkının azaltılması amacıyla bölge planlaması yapılarak bölgesel teşvikler artırılmasına devam edilmiştir (DPT, 2007:32-33). i-Sekizinci Beş Yıllık Kalkınma Planı Bu plan 2001 yılında yürülüğe girerek 2005 yılına kadar devam etmiştir. Bu dönemde kamu sanayi kuruluşlarının özelleştirilmesi çalışmalarına devam edilmiştir. Sanayi sektörünün hammadde enerji talebin sağlamak ve enerji güvenliğini artırmak amacıyla madencilik sektöründeki çalışmalara önem verilmiştir. İmalat sanayinin dışa dönük bir yapı içerisinde rekabet gücünün artırılması planlanmaktaydı. Bilgi ve teknoloji ağırlıklı özelliği olan savunma ve havacılık makine imalat, kimya ve elektronik sanayinin ve alanın ilerletilmesi, sanayide gelişmiş teknolojinin kullanımının artırılması, geleneksel sanayilerin rekabet gücünün artırılması hedeflenmiştir (DPT, 2001:117-123). ii-Onuncu Kalkınma Planı Bu kalkınma planı 2014 yılında yürülüğe girmiş ve 2018 yılına kadarki süre için planlama yapılmıştır. Sanyi sektörünün rekabet gücü artırılarak yurtiçi diğer sektörler ve alt sektörler ile değer zinciri bazlı bütünleşmenin artırılması sağlanacaktır. Sanayi girdilerinde yurtiçi ürün oranı artırılacak, özellikle savunma sanayisinde yerli ürün kullanımına dönük çalışmalar artırılması planlanmıştır. Yeşil ve doğa dostu üretim dönüşümü sağlayıcı 115 Mehmet Kaya / Muhammed Çelik \| 115 politikalar geliştirilmesi hedeflenmiştir. Yerli üretimin inovatif ve yeşil üretim kapasitesini artırmada kamu alımları teşvik edici bir rol üstlenmesi, ortak Ar-Ge ve tasarım, ortak tedarik ve pazarlama faaliyetlerinin geliştirilmesi özendirilmesi hedeflenmiştir. Yeşil üretim kapsamında sanayi sektöründe geri dönüşüm ve geri kazanım gibi uygulamalara önem verileceği belirtilmiştir. Onuncu kalkınma planında sanayi sektörü açısından; inovatif, yeşil, dönüştürülebilir, rekabetçi, yerli girdi artırıcı, ihracat yönü yüksel ve katma değeri yüksek bir üretim sağlanması için politikalar geliştirilmiştir (DPT,2014:87-89). iv-Onbirinci Kalkınma Planı Bu kalkınma planı 2019 yılında kabul edilerek Türkiye’ nin 2023 yılı hadefleri doğrultusunda hazırlanmış bir plandır. Diğer kalkınma planlarından farklı olarak Devlet Planlama Teşkilatının kapatılması ile Türkiye Cumhuriyeti Cumhurbaşkanlığı Strateji ve Bütçe Başkanlığı tarafından hazırlanmıştır. Cumhurbaşkanlığı Hükümet Sistemi tarafından hazırlanan ilk kalkınma planıdır. Hazırlanan bu planda öncelikle sanayi sektörünün gelişimi açısından finans erişimini kolaylaştırıcı hale getirileceği belirtilmiştir. Finansmanda teknoloji tabanlı girişimler önceliklenmesi hedeflenmiştir. Teknoloji Odaklı Sanayi Hamlesi Programı kapsamında öncelikli sektörlerdeki yatırımlar ile ilgili süreçler bir bütün şeklinde ürün odaklı olarak desteklenmesi hedeflenmiştir (Cumhurbaşkanlığı Strateji ve Bütçe Başkanlığı, 2018:58-124). 2005 yılından sonra isekamu hem ve özel sektörün faaliyetleri için yol gösterici olması yanında makro politika ve hedeflerin birlikte sektörel bazdaki gelişimi de kapsayan üç yıl vadeli orta vadeli programlar yürülüğe konulmuştur. Bu programlar itibariyle dönemin sanayileşmeye ilişkin odaklanan hususları tespit emek mümkündür. Nitekim programlarda ekseriyetle;“üretimin ithalata bağımlılığının azaltılması”,”yüksek katma değerli ürün üretilmesi”ar- ge çalışmalarının arttırılması” vb.sanayi sektörünün alışık olduğumuz yapısal sorunları dillendirilmektedir. Belki de plan ve programlar hakkındaki en önemli noktalardan biri de, hedeflerin ne oranda gerçekleştiirldiğine ve gerçekleştirlmemişse sebeplerine ilişkin değerlendirmelerin yapılmasıdır. Yoksa aynı hedefler sürekli tekrarlanmak durumunda kalınacaktır.Son tahlilde sanayide geldiğimiz nokta ilerisi içiin ufuk açıcı olacaktır. 7.Türkiye’ de Sanayi Sektörünün Mevcut Durumu Türkiye Cumhuriyeti’in kuruluşundan önce İzmir İktisat Kongresi’nde kalkınmak,batılılıaşmak ve çağdaş medeniyetler seviyesie ulaşamk üzere belirlenen sanayileşme hedefine ulaşmak için, izlenen liberal milli, devletçilik, 116 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme planlı kalkınma ve son olarak 1980’de başlayan ve bugün de devam eden neoliberal politikların başarısını değerlendirmede kriterimiz, çoğunlukla toplam üretim içinde sanayinin payındaki değişim oldu. Bu bağlamda yirminci yüzyılın son iki yılında, finansal küreselleşme sürecinde çevrede ve ülkemizde yaşanan krizlere rağmen, sanayi sektörünün (İmalat San.,Madencilik, Elektrik, Su, Gaz, ve İnşaat sektörleri) GSYİH’daki payı sırasıyla % 25,4 ve % 23 olmuştur. Yirmibirinci yüzyıla % 21 ile başlayan sanayinin payı uzun yıllar boyunca sürekli, hatta 2004’ten itibaren % 20’nin altına, düşmüş, ancak 2014’te % 20’lere tekrar ulaşırken, yirminci yüzyılın sonundaki değereine 2020 yılını bulmuştur (Tablo 6.). Buna rağmen Türkiye’nin dönem başında % 0,57 olan dünya sanayi üretimindeki ağırlığı % 0,85’e çıkarken, Türkiye sanayi sektörünün uluslararası arenada da konumunu güçlendirerek 30 milyar dolar civarında olan sanayi ihracatı son yıllarda 5 kattan fazla artarak 170 milyar dolara yaklaşmıştır (İstikbal, 2022:9,15). Sanayi üretiminde söz konusu dönem boyunca düşüş yaşanırken, özellikle inşaat ve yanı sıra gayrimenkul faaliyetlerinin GSYİH’da artan payının, döneme damgasını vuran her halukarda büyümeye ve kısa sürede sonuç almaya odaklı popülüst yaklaşımın sonucu olduğunu söylemek mümkündür. Döneme ilişkin yapılabilecek bir başka çıkarsam da Türkiye ekonomisinin hızla hizmet ekonomisi olma yolundaki dönüşümüdür. Ayrıca finans faaliyetlerinin yaklaşık % 3-4 düzeyinde seyreden ağırlığı, yine zamanın ruhuna uygun olarak, ekonominin finansallaşmasının da göstergesidir. İşte sanayileşme hızındaki düşüşün bir nedeni de, reel ekonomide karlılığı düşen sermayenin daha yüksek kar beklentisiyle finansal piyasalara kaymasıdır. Mehmet Kaya / Muhammed Çelik \| 117 Tablo 6. 7.Türkiye’ de Sanayi Sektörünün Mevcut Durumu İktisadi Faaliyet Kollarının GSYİH İçerisindeki Payı (Yüzde) Yıllar Tarım, Ormancılık ve Balıkçılık Sanayi İnşaat Hizmetler Bilgi ve İletişim Finans ve Sigorta Faaliyetleri Gayrimenkul Faliyetleri Mesleki, İdari ve Destek Hizmetleri Kamu Yönetimi,eğitimi İnsan Sağlığ ve Sosyal Hizmet Faaliyetleri Diğer Hizmetler Faaliyetleri Ürün Üzerindeki Vergiler Eksi Sübvansiyonlar 1998 11,8 25,4 6,1 24,2 1,9 5,4 4,7 2,6 8,7 1,3 7,9 1999 10 23 5,6 22,2 2,1 7,2 6,9 2,6 10,4 1,3 8,7 2000 9,6 21,7 5,2 23,6 2,2 5,2 8,4 2,7 9,7 1,4 10,4 2001 8,3 21 4,7 23,7 2,5 6,1 8,6 2,9 10,1 1,3 10,7 2002 9,7 20,3 4,5 24,7 2,6 3,3 8 3,4 10,4 1,5 11,6 2003 9,4 20,3 4,6 24,5 2,5 2,6 8 3,4 10,7 1,7 12,3 2004 9 19,9 5,3 24,8 2,5 2,5 8,2 3,5 10,6 1,5 12,5 2005 8,8 19,8 5,5 24,4 2,7 2,5 8,5 3,6 10,3 1,4 12,5 2006 7,8 19,8 6,2 24,5 2,5 2,1 8,9 3,7 10,3 1,6 12,5 2007 7,2 19,7 6,7 24,2 2,7 2,2 9,6 4,1 10,6 1,7 11,3 2008 7,2 19,4 6,8 24,2 2,7 2,4 9,7 4,2 10,6 1,8 10,9 2009 7,8 18,5 5,6 22,1 2,7 3,8 10,5 4,5 11,9 2 10,7 2010 8,6 18,5 6 21,6 2,5 3 10 4,3 11,6 2 12 2011 7,9 19,8 7,1 21,9 2,3 2,7 9 4,3 11,1 2 11,9 2012 7,4 19,3 7,4 22,4 2,4 2,9 8,7 4,5 11,3 2 11,6 2013 6,4 19,7 8 22,2 2,4 3,1 8,2 4,6 11 2 12,3 2014 6,3 20,1 8,1 22,6 2,4 2,9 8 4,8 11,2 2 11,5 2015 6,6 19,8 8,1 22,5 2,4 3 7,7 5,2 10,9 1,9 11,8 2016 6 19,7 8,5 21,9 2,4 3,3 7,7 5,3 11,6 1,9 11,7 2017 5,8 20,7 8,5 22,6 2,5 3,2 7,2 5,4 10,9 1,9 11,3 2018(r) 5,5 22,3 7,2 23,7 2,5 3 6,8 5,2 11,4 1,9 10,4 2019(r) 6,2 22 5,4 24,3 2,6 3,2 6,6 5,3 12,4 2,1 9,9 2020(r) 6,4 22,7 5,3 22,3 2,8 3,8 6,4 4,7 12,5 2,2 11 2021(r) 5,3 26 5,1 24,3 2,8 3 5,1 4,7 10,9 2,3 10,6 2022(r) 6 26,7 4,9 26,6 2,3 3,4 3,8 4,5 9,2 2 10,6 Kaynak: TÜİK Verileri üzerinden Yazarlar Tarafından Düzenlenmiştir. ablo 6. İktisadi Faaliyet Kollarının GSYİH İçerisindeki Payı (Yüzde) 118 118 \| Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Tablo 7. 7.Türkiye’ de Sanayi Sektörünün Mevcut Durumu Sanayi Sektörünün İstihdam Kapasitesi Çalışan Aralığı Kapasite Raporu Sayısı Toplam Çalışan 2021 2022 Dönemsel Değişim Yüzdesi 2021 2022 Dönemsel Değişim Yüzdesi 1-9 37.195 42.822 15,13 186.680 214.438 14,87 10-49 34.175 37.552 9,88 793.621 868.743 9,47 50-99 6.932 7.471 7,78 486.247 524.138 7,79 100-249 5.207 5.512 5,86 800.308 851.328 6,38 250+ 2.281 2.447 7,28 1.308.799 1.412.833 7,95 Toplam 85.790 95.804 11,67 3.575.655 3.871.480 8,27 Kaynak: (TÜİK, 2022) Tablo 7. Sanayi Sektörünün İstihdam Kapasitesi Türkiye sanayi sektörünün istihdamdaki ağırlığı uzun yıllardır % 20 oranında çakılı kalırken, İstihdamın yükünü de, muhtemelen üretim teknolojilerinin emek yoğun niteliği dolayısıyla, küçük ve orta boy işletmelerin taşıdığı söylenebilir (Tablo 7-8). Tablo 8. Sanayi Sektöründeki İstihdam Verileri (Bin Kişi) Tablo 8. Sanayi Sektöründeki İstihdam Verileri (Bin Kişi) Yıllar Dönem Toplam İstihdam(Kişi) Sanayi (Kişi) Sanayi İstihdan (Oran) 2019 I. Çeyrek 27.335 5.398 19,7 II. Çeyrek 28.221 5.554 19,7 III. Çeyrek 28.482 5.561 19,5 IV. Çeyrek 28.131 5.775 20,5 2020 I. Çeyrek 26.699 5.603 21,0 II. Çeyrek 25.853 5.301 20,5 III. Çeyrek 27.364 5.408 19,8 IV. Çeyrek 26.823 5.670 21,1 2021 I. Çeyrek 27.391 5.955 21,7 II. Çeyrek 28.448 6.077 21,4 III. Çeyrek 29.652 6.200 20,9 IV. Çeyrek 29.815 6.472 21,7 2022 I. Çeyrek 29.418 6.543 22,2 II. Çeyrek 30.765 6.764 22,0 III. Çeyrek 31.160 6.638 21,3 IV. Çeyrek 31.556 6.725 21,3 2023 I. Çeyrek 30.856 6.824 22,1 II. Çeyrek 31.507 6.818 21,6 Kaynak: (TÜİK, 2022) Mehmet Kaya / Muhammed Çelik \| 119 Mehmet Kaya / Muhammed Çelik \| 119 8. Sonuç ve Değerlendirme 8. Sonuç ve Değerlendirme 8. Sonuç ve Değerlendirme İnsanlığın medeniyet yürüyüşünün seyrini üretim konusundaki olanaklarının gelişimi ile paralellik arz ettiğini söyleyebiliriz. Nitekim tarım devrimi ile geçilen üretim ekonomisi ile birlikte, yerleşik hayatın ve yerleşim mekanlarının inşasından başlamak üzere birçok sosyal, siyasal ve iktisadi gelişmeye kapı aralanmıştır. İnsanlığın uzun tarihinde küçük bir zaman dilimine denk gelse de yaklaşık üç asır önce gerçekleşen sanayi devrimi ise, insanlık için her yönüyle bir milat olarak kabul edilebilir. İnsanlığın bugün için sahip olduğu refahı, zenginleşmeyi, modernleşmeyi ve konforu, buhar makinesiyle başlayan ve bugün dijital teknolojilere (yapay zeka, 3D yazıcı vb.) evrilen sanayi devrimlerine borçludur. Kaldı ki bu devrimleri erken yaşayan ya da adapte olan ülkeler de sanayileşmeyi yanı sıra gelişmeyi de başarmışlardır. Türkiye’nin sanayi/teknoloji devrimlerine dahil olması konusundaki tüm söylemlerde birincisini yaşamamış olmasından dolayı her zaman için bir “kaçırılmış tren” metaforu baskın gelmektedir. Buna ilişkin en önemli gerekçe ise, devamı olduğu Osmanlı Devleti’nin bu konudaki ihmalkarlığıdır. Ayrıca yeni kurulan Cumhuriyetin devraldığı kötü ekonomik mirasın ve savaşın yok ettiği ekonomiyle kaçan treni yakalaması olanaksızdı. Buna rağmen Cumhuriyetin batı medeniyetleri seviyesine erişmek için ekonomideki yol haritası da sanayileşme olmuştur zaten olmalıydı da. Çünkü aklın yolu bir hesabı, gelişmeyi sanayileşme ile başaranların izinden gitmek en doğrusuydu. Bu uğurda bir asırdır, devletçilik, planlama veya özel sektör eliyle kalkınma gibi alternatif modeller uygulansa da, tüm modellerin karma ekonomik modelde birleştiğini/ortaklaştığını söyleyebiliriz. Türkiye’de tarihsel süreçte sanayileşmede sağlanan başarının da mu modelin, yoğun olarak uygulandığı,1923-1939 dönemi başta olmak üzere, dönemlerdir Sanayileşmeyi zorunlu kılan bir başka husus, sanayi sektörünün diğer tüm sektörleri etkileme potansiyelidir Bu sektörde yaşanacak her olumlu gelişme diğer sektörleri de dolaylı ya da olarak etkilemektedir. Ayrıca sanayileşmeyle birlikte hem üretimin hem de nüfusun tarımdan sanayiye geçişi söz konusudur. Nitekim Cumhuriyetin ilk yıllarında tarım sektöründe yaşayan nüfusun yıllar geçtikçe sanayi sektörüne doğru bir geçiş yaşadığını görmekteyiz. Bu durum hem sanayi sektöründe yaşanan gelişmeler kadar, sanayi sektörünün teknolojik gelişmeler ile tarım sektörünü desteklemesi ile alakalıdır. Tarımdaki sanayileşme beraberinde tarımdaki emek yoğunluğunu azaltmış ve bu sektördeki emek fazlalığının sanayi sektörüne doğru geçişine neden olmuştur. 120 \| Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 120 Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme Sanayi sektörü Türkiye’ de istihdamı en çok etkileyen sektördür. Dolayısıyla bu sektörü destekleyici politikalar oluşturulmaya devam edilmelidir. İmalat sanayi bu açıdan en büyük payı sağlamaktadır. Bunun yanında Türkiye’ de son yıllarda inanılmaz bir başarı yakalayan savunma sanayisi de katma değeri yüksek bir sanayi alt sektörü olarak ön plana çıkmaktadır. 8. Sonuç ve Değerlendirme Bu sektörün stratejik önemi ve imalat sanayisindeki yüksek etkileme potansiyeli Türkiye açısından bu iki sektöre özel bir yaklaşım gerekmektedir. Bu sektörleri besleyici nitelikli insan gücünün oluşturulup devamlılığın sağlanması hayati önemdedir. Türkiye sanayi sektörü adeta sonu gelmemiş bir başarı hikayesidir. Prangalardan kurtulmak amacıyla cumhuriyet döneminde atılan önemli adımlar ve ekonomik olarak çizilen vizyonu giriş olarak alırsak, hazırlanan planlar, uygulanan politikalar ve Türk sanayicisinin gösterdiği özveriyle oluşturulan sağlam temeller ve bu süreçte yaşanan çalkantılı olaylar Türk sanayi sektörünün gelişimini ve direncini artırmıştır. Özellikle son sanayi devriminin teknolojilerine uyum konusundaki dijital yol haritası gibi çalışmalar bu duyguyu perçinlemiştir. Ancak bunun yanında Türkiye’nin bu konuda yarım kalan hatta ertelenen hikâyesini tamamlaması için Eşiyok (2018:105), bir asırlık Cumhuriyetin yarım asra dayanan tecrübesine referansla, Türkiye gibi geç sanayileşmeye başlayan yarı-sanayileşmiş bir ekonominin üretimde hızlı yapısal dönüşümleri sağlayarak sanayileşmede ilerlemeyi sağlamış ülkeleri yakalaması için en temel koşulunun, kalkınmayı esasa almış devlet ve özel sektörün işbirliği çerçevesinde, planlı bir kalkınma stratejisine ve bu stratejinin en temel bileşenini oluşturan bağlı olduğunu öne sürmektedir. 121 Mehmet Kaya / Muhammed Çelik \| 121 Kaynakça Kaynakça Alpağut, S. (2022). Türkiye’nin Cumhuriyet Dönemi İktisadi Yaklaşımı ve Poli- tikalarının Değerlendirilmesi. 247-259. Ekin Yayınevi. Baştav, L. (2012). Dışa Açık Büyüme Modelinde Sanayİ Politikalarının Gelişi- mi: Türk İmalat Sanayiinin Yapısı ve Rekabet Gücü (1980-2011). ODTÜ Gelişme Dergisi, 303-322. Boratav, K. (2015). Türkiye İktisat Tarihi 1902009. Ankara: İmge Kitabevi. Cem Somel. (2004). Türkiye İktisat Politikaları Tarih. M. Özel içinde, Türkiye Ekonomisi (s. Eskişehir). Açık Öğretim Fakültes. Doğan, S. (2015). Türkiye Ekonomisi. İstanbul: İstanbul Üniversitesi Açık ve Uzaktan Eğitim Fakültesi Yayını. DPT. (1963, Ocak). Birinci Beş Yıllık Kalkınma Planı. Başbakanlık Devlet Matbaası. DPT. (1979). Dördüncü Beş Yıllık Kalkınma Planı 1979-1983. Ankara: DPT 1664. DPT. (1985). Beşinci Beş Yıllık Kalkınma Planı. Devlet Planlama Teşkilatı. DPT. (1990). Altıncı Beş Yıllık Kalkınma Planı (1990-1994). Devlet Planlama Teşkilatı. DPT. (1995). Yedinci Beş Yıllık Kalkınma Planı (1995-2000). Devlet Planlama Teşkilatı. DPT. (2001). Uzun Vadeli Strateji ve Sekizinci Beş Yıllık Kalkınma Planı 2001- 2005. Devlet Planlama Teşkilatı. DPT. (2007). Dokuzuncu Kalkınma Planı (2007-2013). Devlet Planlama Teşkilatı. Eğilmez, M. (2018). Değişim Sürecinde Türkiye . İstanbul: Remzi Kitabevi. Erdem Koç, M. C. (2018). Dünyada ve Türkiye’de Sanayileşme I - Strateji ve. 59(690), 1-26. Mühendis ve Makine. Eşiyok, B. (2018). Erken Sanayisizleşme ve Bir Kalkınma Stratejis, Tasarımının Ana Çizgileri. İktisat ve Toplum Dergisi, 99-103. Evsile, M. (2018, November). Cumhuriyet Döneminde Sanayileşme Faaliyetleri (1923-1950). 10(8), 110-119. History Studies İnternational Journal Of History. Han, E. (1977). 8. Sonuç ve Değerlendirme Türkiye’ de Sanayileşme Süreci ve Stratejisi. Eskişehir: Anadolu Üniversitesi, Eskişehir İktisadi ve Ticari İlimler Akademisi. Hüseyin Akyıldız, Ö. E. (2004). Türkiye Cumhuriyet Dönemi Uygulanan İk- tisat Politikaları. 9(1), 43-62. Süleyman Demirel Üniversitesi İktisadi ve İdari Bilimler Fakültesi Dergisi. İlkin, A. (1970). Türkiyede Sanayi Politikası. İstanbul Üniversitesi İktisat Fakül- tesi Mecmuası, 1-4(30), 378-423. 122 \| Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 122 İnan, A. (1972). Devletçilik İlkesi ve Türkiye Cumhuriyeti’nin Birinci Sanayi Planı 1933. Ankara: Türk Tarih Kurumu Yayınevi. İstikbal, D. (2022). Türkiye’ de Sanayinin Dönüşümü ve Yeni Teknolojik Trendler. (365). İstanbul: SETA \| SİYASET, EKONOMİ VE TOPLUM ARAŞTIRMALARI VAKFI . Kalaycı İrfan, K. M. (2019). Atatürk Döneminde Ağır Sanayi Sorunsalı. Adam Akademi, 307-328. Kalkınma Bakanlığı. (2014). Onuncu Kalkınma Planı (2014-2018). Devlet Planlama Teşkilatı. Kaya, M. (2020). İktisadi Büyüme ve Kalkınma. İ. Kalaycı içinde, Türkiye Eko- nomisi (s. 7-59). İstanbul: Divan Kitap. Kazgan, G. (1988). Ekonomide Dışa Açık Büyüme. İstanbul: Altın Kitaplar Yayınevi. Kazgan, G. (1999). Tanzimattan 21.Yüzyıla Türkiye Ekonomsis. İstanbul: Altın Kitaplar Yayınevi. Keleş, B. (2021). Sanayi Devriminin Osmanlı Ekonomisi Üzerindeki Etkisi ve Cumhuriyet Dönemi Ekonomik Politikalarına Yansımaları (1876- 1938). (86), 373-388. The Journal of Academic Social Science Studies. Kılıçbay, M. (1992). Türk Ekonomisi. Ankara: Kültür Yayınları. Koç, İ. C. (2000). İzmir İktisat Kongresinin Türk Ekonomisinin OLuşumuna Etkileri. Atatürk Dergisi, 145-167. Özçelik Özer, T. G. (2007). Atatürk Dönemi Ekonomi Politikaları. Afyon Koca- tepe Sosyal Bilimler Dergisi, 9(1), 253-266. Pamuk, Ş. (2010). Osmanlı’dan Cumhuriyete Küreselleşme, İktisat Politikaları ve Büyüme. Ankara: Kültür Yayınları. Polatoğlu, M. G. (2017). İkinci Beş Yıllık Sanayi Planı (1938-1942). 6(1), 55- 87. Atatürk Dergisi. Savaş Durmuş, N. K. (2016). Atatürk Dönemi Türkiye Ekonomisi (1923- 1938). 7(12). Kafkas Üniversitesi İktisadi ve İdari Bilimler Fakültesi KAÜİİBFD. Soyak, A. (1999). Planlı Dönemde Sanayileşme. O. Baydar içinde, 75 Yılda Çarklardan Chip’lere (s. 167-181). İstanbul: Tarih Vakfı Yayınları. Şahin, H. (2006). Türkiye Ekonomisi. Bursa: Ezgi Yayınları. Şahin, H. (2010). Türkiye Ekonomisi. Bursa: Ezgi Yyayınevi. T.C. Cumhurbaşkanlığı, S. (2015). Ekonomik ve Sosyal Göstergeler. Ankara: T.C. Kalkınma Bakanlığı. TMMOB, M. M. (2009). Türkiye’de Kalkınma ve İstihdam Odaklı Sanayileşme İçin Planlama Önerileri. Ankara: Makine Mühendisleri Odası. Mehmet Kaya / Muhammed Çelik \| 123 123 Toprak, M. (1999). Cumhuriyet Dönemi Türkiye Ekonomisi 75 Yıl Armağanı. Kara Harp Okulu Bsımevi: Ankara. Toprak, M. D. (2001). Küreselleşen Dünyada Türkiye Ekonomisi: Serbest Piyasa Devriminin Serüven. Ankara: Siyasal Kitabevi. TÜİK. (2010). İstatistiki Göstergeler. Ankara: TÜİK. Uzunkaya, S. Ş. (2020). Türkiye Ekonomisinde 19291977 Yılları Arasında- ki Sanayi ve Kalkınma Planlarının Değerlendirilmesi. 124 \| Türkiye’nin Yarım Kalan Hikayesi: Sanayileşme 1 Doç. Dr., Anadolu Üniversitesi, İktisadi ve İdari Bilimler Fakültesi, ayazici@anadolu.edu.tr, ORCID ID: 0000-0002-7491-8709 Ayla Yazıcı1 Özet Dünyada çalışan nüfusun yaklaşık %61’i kayıt dışı olarak istihdam edilmektedir. Tüm ülkelerde görülebilen kayıt dışı istihdam özellikle gelişmekte olan ülkelerde oldukça yüksektir. İstihdam faaliyetlerinin resmi belgelere yansıtılmaması ya da gerçekleştirilen istihdamın süresi ve alınan ücretin resmi makamlara eksik veya hiç bildirilmemesi olarak kısaca tanımlayabileceğimiz kayıt dışı istihdam, Türkiye’nin de yıllardır karşı karşıya olduğu önemli sorunlardan biridir. Kayıt dışılığın, çalışma yaşamındaki temel ilkeler ve haklar, sosyal koruma, insana yakışır çalışma koşulları, yapılan işe göre adil ücretlendirme gibi işçi hakları üzerinde olumsuz birtakım etkileri vardır. Türkiye nüfusunun yaklaşık yarısını oluşturan kadınların azımsanmayacak bir oranının kayıt dışı olarak çalışması, ülkenin sosyal, kültürel ve ekonomik yönden gelişmesi önündeki engellerden biri olarak görülebilir. 8. Sonuç ve Değerlendirme İstanbul Ticaret Üniversitesi Sosyal Bilimler Dergisi(Özel Sayı), 14-30. Yakup Kepenek, N. Y. (2003). Türkiye Ekonomisi. İstanbul: Remzi Kitabevi. Yeldan, E. (2003). Küreselleşme Sürecinde Türkiye Ekonomisi. İstanbul: İletişim Yayınları. Yenal, O. (2010). Cumhuriyet’in İktisat Tarihi. İstanbul: Kültür Yayınları. Yıldırm, S. (2006). Türkiye’de 24 Ocak 1980 öncesi ve sonrası sanayileşme ve ekonomik büyümeye etkileri. Eskişehir Osmangazi Üniversitesi Sosyal Bilimler Dergisi. Yücel, F. (2014). Cumhuriyet Türkiye’ sinin Sanayileşmede İlk Önemli Adımı: Birinci Beş Yıllık Sanayi Planı 1934-1938. Ankara: TMMOB Elektrik Mühendisi Odası. Yücel, H. (2017). Cumhuriyet Ekonomisinin Kuruluşu Gelişimi. Ankara: Türkiye Barolar Birliği Yayınları. Yülek Murat, M. H. (2020). Sanayi ve Sanayileşme. İ. Kalaycı içinde, Türkiye Ekonomisi (s. 261-305). İstanbul : Duvar Kitap. Bölüm 8 Türkiye’de Kadınların Kayıt Dışı İstihdamı Türkiye’de Kadınların Kayıt Dışı İstihdamı 1. Giriş Kayıt dışı ekonomi günümüzde özellikle gelişmekte olan ülkeler olmak üzere tüm ülkelerin karşı karşıya oldukları en önemli sorunlardan biridir. Dünyadaki çalışan nüfusun tahmini olarak yaklaşık %60’ı kayıt dışı çalışırken, tüm kayıt dışı çalışanların %90’ı gelişmekte olan ülkelerde yoğunlaşmıştır. Ancak kayıt dışı istihdamın boyutunun kesin olarak belirlenmesi neredeyse imkansızdır. Her ne kadar kayıt dışı sektörün büyüklüğünü belirlemek amacıyla çeşitli çalışmalar yapılmış olsa da gerek kullanılan yöntemlerdeki farklılıklar nedeniyle gerek formel ve enformel sektörlerin ayrılmaz bir biçimde iç içe geçmiş olması nedeniyle farklı tahminlere ulaşılmıştır. Kayıt dışı ekonomi, dünya genelindeki toplumların ekonomik kalkınmanın önündeki en büyük zorluklardan biridir. Kayıt dışı çalışanları kayıtlı ekonomideki işçilerden ortalama olarak daha az gelir elde etmektedirler ve https://doi.org/10.58830/ozgur.pub401.c1756 125 Türkiye’de Kadınların Kayıt Dışı İstihdamı 126 aynı zamanda daha az üretkendirler. Dolayısıyla kayıt dışılığın (hem nedenleri hem de sonuçları) ele alınması, sürdürülebilir ve kapsayıcı ekonomik büyüme ve kalkınmayı teşvik etmeye yönelik her türlü stratejinin ayrılmaz bir bileşenidir. Kayıt dışı istihdamın en temel özelliği sosyal koruma kapsamı dışında kalması dolayısıyla güvencesiz bir istihdam olmasıdır (Parlak ve Çiftçi, 2022:98). Türkiye’de de son yıllarda kayıt dışı istihdamda bir düşüş meydana gelmiş olmasına rağmen yine de Uluslararası Çalışma Örgütü 2022 verilerine göre toplamda %28,1 oranında ve TÜİK verilerine göre de %23,4 gerçekleşmiştir2. Büyüklüğü, nedenleri ve etkileri ülkelere göre farklılık gösterse de kayıt dışı ekonomideki artış ülkenin milli gelir, sivil işgücü, işsizlik, tüketim, enflasyon, büyüme oranı, dış ticareti gibi ekonomik göstergelerin güvenirliliğini kaybetmesine neden olduğundan, kayıt dışı ekonomi ve kayıt dışı istihdam ile mücadele önem taşımaktadır. Az gelişmiş, gelişmekte ve geçiş ekonomileri ve hatta gelişmiş ülkelerde dahi kayıt dışı ekonominin varlığı 1990’lı yıllardan bu yana giderek artış göstermiştir. Bu nedenle kayıt dışı ekonominin kayıt altına alınması bütün ülkeler açısından çözümlenmesi gereken önemli bir sorundur. Bütün ülkelerde özellikle kadınların erkeklere oranla daha fazla kayıt dışında istihdam edilmeleri, kadınların birçok haklardan mahrum kalmalarında neden olarak, mağdur olmalarına yol açmaktadır. 2 Elde edilen rakamlardaki farklılık kullanılan ölçe yöntemlerindeki farklılıktan kaynaklanmaktadır. 2. Kayıt Dışı İstihdam Tanımı ve Özellikleri Kayıt dışı ekonomi, sıklıkla birbirinin yerine kullanılan iki istatistiksel terim ve tanımı karşılayan bir kavramdır. Bunlardan ilki, bir işletmenin vergi amaçlı kayıtlı olup olmadığını ifade eden “kayıt dışı sektördür”. İkincisi hem kayıt dışı sektör içinde hem de dışında sosyal ve hukuki korumadan yoksun istihdamı ifade eden “kayıt dışı istihdamdır”. Dolayısıyla “kayıt dışı ekonomi”, bu şekilde tanımlanan tüm faaliyetlere, çalışanlara ve bunlardan elde edilen çıktılara işaret etmektedir. İlk kez 1971 yılında Keith Hart tarafından kayıtlı sektörün dışında çalışan işgücünün bir kısmını tanımlamak için kullanılan kayıt dışı ekonomi kavramı (Shady, 2018:5), 1972 yılında Uluslararası Çalışma Örgütü’nün düzenlediği Dünya İstihdam Programı kapsamındaki Kenya Raporu’nda “enformal sektör” olarak kullanılmıştır. Uluslararası Çalışma Örgütü (ILO) kayıt dışı ekonomiyi, “kanunen veya uygulamada resmi düzenlemeler kapsamında yer 127 Ayla Yazıcı Ayla Yazıcı almayan veya yeterince kapsanmayan, işçiler ve ekonomik birimler tarafından gerçekleştirilen ekonomik faaliyetler” şeklinde tanımlamaktadır. Uluslararası Çalışma Örgütü (ILO), Kayıt Dışı Sektör İstatistikleri Uluslararası Uzman Grubu (“Delhi Grubu”) ve küresel eylem-araştırma- politika ağı olan Kayıt Dışı İstihdamda Kadınlar: Küreselleşme ve Organizasyon (WIEGO), kayıt dışılığın tüm boyutlarını kapsaması için hem kayıt dışı işletmelerin hem içindeki hem de dışındaki kayıt dışı istihdamı kapsayan genişletilmiş kavram ve istatistiksel tanımını geliştirmek için birlikte çalışmışlardır. Bu genişletilmiş tanım, 2002 yılında Uluslararası Çalışma Konferansında (ILC) ve 2003 yılında Uluslararası Çalışma İstatistikçileri Konferansında (ICLS) onaylanmıştır (Chen, 2016:1). 1993 yılında ICLS tüzel kişiliği olmayan küçük ve/veya kayıtlı olmayan işletmelerde gerçekleşen istihdam ve üretime atıfta bulunmak üzere kayıt dışı istihdamın uluslararası istatiksel tanımını benimsemiştir. 2003 yılında ise ICLS, bu tanımı, kayıt dışı işletmelerin dışındaki çeşitli kayıt dışı istihdam türlerini de kapsayacak şekilde genişletmiştir ve bu daha geniş kavrama, kayıt dışı istihdam denmiştir (Chen, 2012:7). ICLS’nin kılavuz ilkelerinde kayıt dışı istihdamı tanımlamak için “Çalışanların, istihdam ilişkileri kanunen veya uygulamada ulusal iş mevzuatına, gelir vergisine, sosyal güvenlik korumasına veya istihdam hakkına tabi değilse, kayıt dışı işlerde çalıştıkları kabul edilir” ifadesi yer almaktadır (ICLS, 2003:2). Bu tanımdan hareketle kayıtlı ve kayıt dışı istihdam, istihdam durumuna göre kayıt dışı işletmelerde serbest meslek ve kayıt dışı ücretli istihdam şeklinde daha homojen alt sektörlere göre sınıflandırmak mümkündür. Kayıt dışı işletmelerde serbest meslek, işverenler ve kendi hesabına çalışan işletmeciler de dahi olmak üzere, kayıtsız veya tüzel kişiliği olmayan işletmelerdeki işçiler (buna aile işletmelerinin başkanları da dahildir), ücretsiz aile çalışanları ve kayıt dışı üretici kooperatiflerinin üyelerini kapsamaktadır (Chen, 2005:38; Shady: 2018:5; ILO, 2018: 9; ICLS, 2003: 13). 2. Kayıt Dışı İstihdam Tanımı ve Özellikleri Kayıt dışı ücretli istihdam ise, kayıtlı veya kayıt dışı işletmeler tarafından sosyal güvence haklarına tabi olmaksızın veya haneler tarafından ücretli ev işçisi olarak işe alınan çalışanları kapsamaktadır. Bazı ücretli iş türlerinin kayıt dışı olma olasılığı diğerlerine göre daha yüksektir ve kayıt dışı işletmelerin çalışanlarını, gündelik işçileri, geçici veya yarı zamanlı çalışanları, ücretli ev işçileri, sözleşmeli çalışanları, kayıtsız veya beyan edilmeyen çalışanları ile ev çalışanlarını kapsamaktadır (Chen, 2005:38; Shady: 2018:5; ILO, 2018: 9; ICLS, 2003: 13). Yapılan bu tanımlamalardan da çıkartılabileceği gibi, kayıt dışı istihdam, istihdam faaliyetlerinin çalışılan gün ve ödenen ücretin vergi dairesi veya Türkiye’de Kadınların Kayıt Dışı İstihdamı 128 sosyal güvenlik kurumuna eksik veya hiç bildirilmemesini ifade etmektedir. Bunun dışında ücretli çalışanların yanı sıra, işverenlerin ve kendi hesabına çalışanların da kayıt dışı istihdamda yer almaları söz konusu olmaktadır (Yanıcı Erdal, 2019: 228). Kayıt dışılık genellikle çalışanlar ve işletmeler için daha büyük mali risklere veya çabalarının karşılığında daha düşük getirilere yol açabilmektedir. Özellikle kayıt dışı çalışma ve aile işçiliğine katkıda bulunma olasılıklarının daha yüksek olduğu düşük gelirli ülkelerde, kadınlar için ek zorluklar ortaya çıkarabilir; bu durum, düşük ücretle veya hiç ücret alınmadan çalışma olarak nitelendirilebilir. Kayıt dışı istihdam kişilerin sosyal korumanın, çalışma haklarının ve insana yakışır çalışma koşullarının eksikliğinin olduğu bir ortamda çalışmalarına neden olmasının yanı sıra, gelir ve geçim kaynakları ile ilgili ciddi belirsizlikler ile karşı karşıya kalmalarına da neden olmaktadır. Özellikle yakın geçmişte yaşanan COVİD-19 krizi, kayıt dışılıkla ilgili önceden de var olan yapısal zayıflıkların daha da artmasına neden olmuştur. Gerek bireylerin gerekse işletmelerin kayıt dışında faaliyet göstermelerinin farklı nedenleri olabilmektedir. Bazıları için kayıtlı bir iş olanağının olmaması durumunda hayatta kalma stratejisi olurken, başkası için amaçlı, kişisel çıkarlarını maksimize etme yönündeki bir karardır, çünkü bazen kayıt dışılık daha fazla esneklik ve bağımsızlık sunabiliyor. Bazı kişiler veya işletmeler kayıtlı ekonominin düzenlemelerine ve yasalarına uymanın maliyetinin, getirisinden fazla olduğunu düşündüklerinden, kendilerine rekabet avantajı sağladığı için kayıt dışında kalmayı tercih etmektedirler. Kayıt dışı istihdam, turizm, tarım, imalat ve birçok farklı hizmet türlü dahil olmak üzere çok çeşitli sektörlerde vardır. Her ne kadar kayıt dışı ekonominin bazı temel özellikleri benzerlikler taşısa da belirli özellikleri ve zorlukları ile bununla mücadele yöntemleri ülkelere özgü olmaktadır. Bir ülke için uygun olan politika çözümleri mutlaka başka bir ülkeye aktarılıp kullanılamayabilir. Bu nedenle kayıt dışılığın faktörleri, özellikleri, nedenleri ve koşulları ulusal bağlamda değerlendirilerek, kayıt dışı ekonomiden kayıtlı ekonomiye geçişi kolaylaştırmak için ülkeye özgü tutarlı ve entegre stratejilerin (kanunlar, düzenlemeler, politikalar ve diğer önlemler de dahil) tasarlanması gerekmektedir. 3. Kayıt Dışı İstihdamın Nedenleri Kayıt dışı ekonomi, birden fazla etkenin ve ulusal koşullara bağlı değişkenliğin söz konusu olduğu karmaşık bir sorundur. Akademik literatürde kayıt dışılığın nedenleri konusunda iki ana düşünce ekolü bulunmaktadır. 129 Ayla Yazıcı Ayla Yazıcı Birincisi, kayıt dışılığın az gelişmişliğin bir belirtisi olduğu görüşündedir. Bu yaklaşıma göre kayıt dışı ekonomi, çalışanların ve firmaların düşük üretkenliğinin bir sonucudur. Bunun nedeni fiziksel sermaye eksikliği, düşük eğitim düzeyi ve bazı sosyo-demografik faktörler (genç ve kırsal nüfusun yüksek olması gibi) gibi yapısal faktörlerdir. Bu yaklaşıma göre kayıt dışılığın azaltılmasına yönelik politikaların işgücü ve iş verimliliğini artırmaya yönelik olması gerekmektedir. İkinci düşünce ekolü, kayıt dışılığın kötü yönetimin bir sonucu olduğunu savunmaktadır. Dolayısıyla kayıt dışılık, özel sektörün aşırı düzenlenmiş bir ekonomiye ve verimsiz bir Devlete tepkisidir. Bu yaklaşıma göre firmalar, düzenlemelere uymanın maliyetinin resmi şirketlere sunulan kamu hizmetlerine erişimden elde edilecek faydayı aştığı durumlarda kayıt dışı sektörde faaliyet göstermeyi seçmektedir. Eğer kayıt dışılık kötü yönetimin bir sonucuysa, politikalar düzenleyici ortamın ve kamu hizmetlerinin iyileştirilmesine odaklanmalıdır. Her iki düşünce okulu da önemli iç görüler sunmaktadır. Kayıt dışı ekonomi karmaşık ve çok yönlü bir olgudur, tek bir nedene bağlanabilecek bir olgu değildir. Kayıt dışı ekonominin nedenleri mali, ekonomik, politik, hukuki, sosyal ve psikolojik etkenlere bağlı olarak ülkeden ülkeye farklılık göstermektedir. Hatta aynı ülke içerisinde yıllara göre de değişim gösterebilen kayıt dışı ekonomi bir ülkenin makroekonomik göstergelerine, ekonomik istikrarına ve gelişmişlik düzeyine bağlıdır (Erdinç, 2016:40). Kayıt dışı sektörün kayıtlı ekonomiye entegrasyonunu engelleyen çok sayıda engel, kayıt dışı ekonominin kapsamının da aynı zamanda artmasına yol açmaktadır. Bunlar arasında işletme kurma, finansman sağlamanın yüksek maliyetleri ve karmaşık prosedürleri yer almaktadır. Ayrıca yetersiz eğitim ve öğretim politikalarının uygulanması, yüksek işlem maliyetlerinin, resmi ekonomide özellikle emek piyasalarındaki artan düzenlemelerin, yolsuzluğa ve verimsizliğe eğilimli idari sistemlerin varlığı, yüksek vergi yükünün bulunması, mülkiyet haklarının olmayışı, krediye erişim eksikliğinin olması ve istikrarlı bir hukuk ve yargı sisteminin olmayışı gibi nedenler de bu entegrasyonun önündeki engeller olarak sayılabilir. Bunun dışında vergi ve sosyal güvenlik katkı paylarındaki artış, işgücü piyasasındaki artan düzenlemeler, hızla artan kentleşme, erken emeklilik, haftalık çalışma saatlerindeki zorunlu düşüşler ve özellikle kadınların kayıtlı sektörlerdeki işlere erişimini sınırlayan güçlerin olması da kayıt dışı istihdamın büyümesine neden olan faktörler arasında yer almaktadır. 130 Türkiye’de Kadınların Kayıt Dışı İstihdamı 4. Türkiye’de Kayıt Dışı İstihdam Türkiye’de özellikle 1980’li yıllardan beri kayıt dışı istihdam yüksek bir seyir izlemektedir. İşsizliğin ve yoksulluğun yüksek olması Türkiye’de kayıt dışı istihdamın en belirgin nedenleri arasında yer almaktadır. Bu nedenle gelirlerini arttırmak ve yaşam standartlarını iyileştirmek için kayıtlı sektörde iş bulamadıklarında kayıt dışı çalışmayı tercih etmektedirler veya ikinci bir işte kayıt dışı çalışmaktadırlar. Kayıt dışı işgücü piyasasının büyümesi, arz ve talep yönlü faktörlerin eş zamanlı hareketinden kaynaklanmaktadır. Talep tarafında Türkiye’nin küresel pazarlara entegrasyonunun artması, ticarete konu olan sektörlerde fiyat rekabetini ve üretim maliyetlerinin en aza indirilmesine yönelik baskıların yoğunlaşmasına neden olmuştur. Bu durum da kayıt dışı istihdamında talep odaklı bir artışa yol açmıştır. Arz tarafında ise, özelleştirme ve kamu sektörü istihdamının azalan payı ile birleşen kitlesel kırdan kente göç modelleri, kayıt dışı istihdam için ilave bir iş gücü sağlamıştır (Aydın et.al., 2010:1). Kayıt dışı istihdamı arttıran diğer faktörler olarak, yüksek işsizlik oranı, ağırlıklı olarak küçük ölçekli işletmelerin faaliyet göstermesi, yüksek enflasyon oranı, yüksek istihdam vergileri gibi unsurlar da sayılabilmektedir. Ayrıca eğitim düzeyinin düşük olması ve nitelikli işgücünün yetersizliği de bireyleri kayıt dışında çalışmaya itmektedir. Ancak kayıt dışı çalışma nedeniyle ülke ekonomilerinin gelir kalemlerinde bir azalma meydana gelmektedir, çünkü kayıt dışında çalışanların sosyal güvenlik hakkından yararlanmamaları nedeniyle, ücret üzerinden yasal vergi, sosyal güvenlik primleri gibi kesintiler gerçekleşmemektedir. Dolayısıyla kamu kurumlarına ödenmesi gereken bu kesintiler ödenmediğinden, devlet gelirlerinde bir azalma meydana gelmekte ve ülke ekonomisi bundan olumsuz etkilenmektedir. Ayrıca sosyal güvenlik haklarından yararlanamayan bu bireyler çeşitli sorunlar ile karşı karşıya kalabilmektedir. 131 Ayla Yazıcı Şekil 1: 2021 Yılında Türkiye ve OECD Ülkeleri İşgücüne Katılım, İstihdam ve İşsizlik Oranları Dünya genelinde gerçekleşen kayıt dışı istihdam oranına göre daha düşük 133 Ayla Yazıcı Ayla Yazıcı olmasına rağmen, halen özellikle gelişmiş ülkeler karşısında yüksek olduğunu ve Türkiye ekonomisini olumsuz yönde etkilemeye devam ettiği söylenebilir. ILO tarafından 2018 yılında yapılan araştırmaya göre, dünya genelinde %61 oranında gerçekleşen kayıt dışı iştihamın gelişmekte olan ülkelerde %90 ve gelişmiş ekonomilerde %18 olduğu belirtilmektedir (ILO, 2018: 13). Şekil 3: 2000-2022 Yılları Arasında Türkiye’de Kayıt Dışı İstihdamın Seyri Kaynak: International Labour Organization. ilostat.ilo.org/data Kaynak: International Labour Organization. ilostat.ilo.org/data Şekil 1: 2021 Yılında Türkiye ve OECD Ülkeleri İşgücüne Katılım, İstihdam ve İşsizlik Oranları Şekil 1: 2021 Yılında Türkiye ve OECD Ülkeleri İşgücüne Katılım, İstihdam ve İşsizlik Oranları Kaynak: OECD (2023). stats.oecd.org Kaynak: OECD (2023). stats.oecd.org Türkiye ve Ekonomik İşbirliği ve Kalkınma Örgütü (OECD) ülkelerine ait 2021 yılı işgücüne katılım, istihdam ve işsizlik oranlarının verildiği Grafik 1’e bakıldığında hem toplam hem cinsiyete göre işgücü katılım oranı ve toplam istihdam oranının OECD ülkelerinin ortalamasının altında kaldığı görülmektedir. Buna karşılık işsizlik oranının yaklaşık iki kat daha fazla olması göze çarpmaktadır. Özellikle kadınların işgücüne katılım oranı ve istihdam oranı gerek Türkiye’deki erkeklere göre gerek OECD ülkelerindeki hemcinslerine göre çarpıcı bir nitelikte düşük kaldığı görülmektedir. OECD’ye üye ülkeler arasında Türkiye’de kadınların istihdama katılım oranı ve istihdam oranı en düşük seviyede kalmaktadır. Bu da Türkiye’de kadınların özellikle kültürel normların etkisi altında kalarak işgücü piyasasında kendilerine yer bulmadıklarını göstermektedir. Türkiye’de kadınların işgücü piyasası dışında kalmalarının nedenleri arasında, kadınlara toplum ve aile içerisinde yüklenen geleneksel roller, kadının ev dışında çalışmasının değerli görülmemesi, bazı meslek ve iş alanlarının kadınlar için uygun kabul edilmemesi gibi cinsiyetçi yaklaşımlar sayılabilir. Ayrıca, bazı işyerlerinde kadının çalışması için uygun ortam ve koşulların sağlanmaması (kreş vb.), doğum ve çocuk bakımı gibi nedenlerle kadının işini aksatacağı yönünde önyargıların bulunması, bazı bölgelerde 132 Türkiye’de Kadınların Kayıt Dışı İstihdamı kadınların eğitim seviyesinin daha düşük olması gibi nedenler de sıralanabilir (İstanbul Kalkınma Ajansı, 2015:3). Her ne kadar son yıllara göre kadınların işgücüne katılma oranı ve istihdam oranı artması, işsizlik oranının da düşmesi sevindirici olsa da halen Türkiye’de 21 milyonun üzerinde kadının herhangi bir gelire sahip olmadan başkalarına bağlı bir şekilde hayatını sürdürmeye çalışması hem üzücü hem düşündürücüdür. Kadınların büyük bir kısmının işgücü piyasasının dışında kalması ayrıca ekonomide kaynak dağılımı sorununu da beraberinde getirmektedir ve kadınların işgücü piyasasındaki konumunun önemini ortaya koymaktadır. 2022 yılında kadınların tarım ve tarım dışı alanlardaki ve işteki durumlarının verildiği şekil 2’ye bakıldığında, tarım sektöründe çalışan kadınların yarısından fazlasının ücretsiz aile işçisi olarak çalıştıkları görülebilir. Tarım dışında çalışan kadınların ise 6 milyondan fazlası hizmetler sektöründe istihdam edilmektedir. Burada da toptan ve perakende ticaret, eğitim ve bakım hizmetlerinde ağırlıklı olarak çalıştıkları görülmektedir (TÜİK, İşgücü İstatistikleri 2023). Şekil 2: 2022 Yılında İstihdamdaki Kadınların Sektörlere ve İşteki Durumlarına Göre Sayıları (bin) Kaynak: TÜİK, İşgücü İstatistikleri 2023. Kaynak: TÜİK, İşgücü İstatistikleri 2023. 2000-2022 yılları arasında Türkiye’de kayıt dışı istihdamının seyrinin verildiği şekil 3’e bakıldığında, kayıt dışı istihdamın düştüğü görülmektedir. 5. Türkiye’de Kadınların Kayıt Dışı İstihdamı Kadınların istihdamda yer alması ülkenin gelişmişlik düzeyi açısından en önemli göstergelerden biridir. Kayıtlı sektörde cinsiyete dayalı olarak birçok ayrımcılıkla karşılaşan kadınlar kayıt dışı sektörde yoğunlaşmaktadırlar. Kırsal kesimde ücretsiz aile işçisi olarak çalışırken, kentsel kesimde ise ev hanımı olarak ve ev işlerinde çalışanlar olarak (çocuk bakımı, temizlik işçiliği, ev işçiliği gibi) faaliyetlerini sürdüren kadınlar, böylece kayıt dışı istihdamın artmasına neden olmaktadırlar (Şahin ve Develi, 2021: 388). Çeşitli ülkelerin 2022 yılına ait kadınların istihdam oranlarının verildiği Tablo 1’den de görüldüğü gibi, gelişmiş ekonomilerde kadınların istihdam oranı gelişmekte olan ülkelere kıyasla daha yüksektir. Almanya, ABD, Fransa, Japonya gibi gelişmiş ekonomilerde kadınların istihdam oranı %40’ın üzerindeyken, Türkiye’de bu oran %32,7 olarak gerçekleşmiştir. Diğer taraftan İran, Irak, Pakistan ve Mısır gibi Müslüman ülkelere bakıldığında, kadınların istihdam oranın çok düşük seviyelerde gerçekleştiği görülmektedir. 134 Türkiye’de Kadınların Kayıt Dışı İstihdamı Erkek egemenliğinin daha fazla olduğu bu ülkelerde, dini, toplumsal ve aile içi baskılar kadınları işgücü piyasasının dışında tutmaktadır. Tablo 1: 2022 Yılında Çeşitli Ülkelerde Kadınların İstihdam Oranları (%) Ülke İstihdam Oranı (%) ABD 46.4 Almanya 46.7 Avusturya 46.7 Bulgaristan 46.4 Çin 45.2 Fransa 48.8 Hindistan 23.5 Irak 14.4 İran 18.3 İsveç 47.4 Japonya 44.8 Kanada 47.4 Mısır 18.0 Pakistan 23.3 Türkiye 32.7 Yunanistan 44.7 Kaynak: Dünya Bankası, 2023. Tablo 1: 2022 Yılında Çeşitli Ülkelerde Kadınların İstihdam Oranları (%) Ülke İstihdam Oranı (%) ABD 46.4 Almanya 46.7 Avusturya 46.7 Bulgaristan 46.4 Çin 45.2 Fransa 48.8 Hindistan 23.5 Irak 14.4 İran 18.3 İsveç 47.4 Japonya 44.8 Kanada 47.4 Mısır 18.0 Pakistan 23.3 Türkiye 32.7 Yunanistan 44.7 Kaynak: Dünya Bankası, 2023. Tablo 1: 2022 Yılında Çeşitli Ülkelerde Kadınların İstihdam Oranları (%) Kaynak: Dünya Bankası, 2023. Kaynak: Dünya Bankası, 2023. Dünya genelinde erkekler (%63,0) kadınlara (%58,1) göre daha yüksek oranlarda kayıt dışı istihdam edilirken, düşük gelirli ülkelerde tam tersi olduğu görülmektedir. (ILO, 2018: 13). Türkiye’deki boyutuna bakıldığında ise, tablo 2’den de görülebileceği gibi, kayıt dışı istihdam kadınlar arasında daha yaygındır. TÜİK tarafından yayımlanan verilere göre, her ne kadar yıllar itibariyle kadınların kayıt dışı istihdam oranları düşme eğilimi göstermiş olsa da erkeklere kıyasla halen yüksek boyutlardadır. Özellikle tarım sektöründe kadınların kayıt dışı istihdam oranının sürekli %90’nın üzerinde seyretmesi, tarımsal alanda faaliyet gösteren kadınların neredeyse tamamının kayıt dışı olarak çalıştığını göstermektedir. Türkiye’de kadınların kayıt dışı istihdam oranının bu denli yüksek olması, Türkiye gibi gelişmekte olan bir ülkede kadınlara biçilen toplumsal cinsiyet rolü sonucunda, kadınların daha ziyade ev ve bakım hizmetlerini yerine getirmeleri yönündeki toplumsal ve kültürel algıdan kaynaklandığını söylemek mümkündür (Bayar, 2021: 8). 3 Ev eksenli çalışma, ILO’nun 1996 tarihli ve 177 sayılı Ev Eksenli Çalışma Sözleşmesi ve eşlik eden 184 sayılı Tavsiye Kararı’nda şöyle tanımlanmaktadır: “… bir kişi tarafından yürütülen ve (i) işverenin işyeri dışında kalan, kişinin kendi evi veya kendi seçtiği diğer tesislerde, (ii) ücret karşılığında, (iii) donanım, malzeme veya kullanılan diğer girdileri kimin sağladığına bakılmaksızın işveren tarafından belirlenen bir ürün veya hizmet ile sonuçlanan çalışmadır” (177 sayılı Sözleşme, 1. Madde). Uluslararası Çalışma Örgütü (ILO). (2021). Working from Home: From invisibility to Decent Work. Geneva. 5. Türkiye’de Kadınların Kayıt Dışı İstihdamı Bunun dışında kayıt dışı çalışmanın (çocuk bakımı, temizlik işçiliği, ev eksenli çalışma3) esnek çalışma koşulları sağlaması nedeniyle kadınlar tarafından tercih de edilebilmektedir. 135 Ayla Yazıcı Tablo 2: Türkiye’de Yıllar İtibariyle Cinsiyete Göre Tarım ve Tarım Dışı Kayıt Dışı İstihdam Oranları (%) Erkek Kadın Yıllar Toplam Tarım Tarım Dışı Toplam Tarım Tarım Dışı 2014 29.1 71.2 21.3 47.5 94.1 26.0 2015 28.1 69.6 20.4 45.6 93.8 24.4 2016 28.9 71.6 21.0 44.4 94.1 24.5 2017 29.4 73.8 21.3 44.5 94.1 25.2 2018 29.6 75.9 21.5 42.3 90.8 25.3 2019 31.1 78.9 22.8 42.5 95.6. 24.7 2020 27.9 76.0 19.2 37.5 94.3 20.6 2021 25.9 77.7 17.0 35.9 94.2 18.7 2022 23.4 72.6 15.7 33.8 90.1 19.2 Kaynak: TÜİK, 2023. Tablo 2: Türkiye’de Yıllar İtibariyle Cinsiyete Göre Tarım ve Tarım Dışı Kayıt Dışı İstihdam Oranları (%) Kaynak: TÜİK, 2023. Şekil 4: 2021 Yılına Göre Çeşitli Ülkelerde Kadınların Kayıt Dışı Oranları Kaynak: TÜİK, 2023 ve International Labour Organization. ilostat.ilo.org/data Şekil 4: 2021 Yılına Göre Çeşitli Ülkelerde Kadınların Kayıt Dışı Oranları Kaynak: TÜİK, 2023 ve International Labour Organization. ilostat.ilo.org/data 136 Türkiye’de Kadınların Kayıt Dışı İstihdamı Türkiye’de kayıt dışı istihdam oranının yıllar itibariyle kayda değer bir azalma göstermektedir. Ancak bazı gelişmekte ve gelişmiş ülkelere göre kadınların kayıt dışı istihdam oranlarının karşılaştırılmalı olarak veren şekil 4’e bakıldığında, toplam ve tarım dışı kayıt dışı oranının Türkiye’de gelişmekte olan ülkelere kıyasla daha düşük olmasına rağmen, gelişmiş ekonomilerle karşılaştırıldığında halen çok yüksek boyutlarda olduğu görülmektedir. Buna karşılık kadın istihdam oranının da gelişmiş ülkelere kıyasla düşük olan gelişmekte olan ülkelerde kadınların özellikle tarım sektöründe ücretsiz aile işçisi olarak veya yevmiye usulü kayıt dışı çalıştıkları söylenebilir. Bu koşullarda çalışan kadınların sosyal güvenceleri olmadığından gerek emeklilik ve gerekse işsizlik sigortası gibi haklarından yararlanamamaktadırlar. Bunun dışında sanayi kesiminde üretim yapan firmalar ulusal ve uluslararası rekabete ayak uydurabilmek için kadınları ucuz işgücü olarak görmektedirler (Yenilmez, 2023:139). Özellikle fason imalat ile kadınların daha düşük ücretlerle ev eksenli olarak kayıt dışı istihdam edilmelerine yol açmaktadırlar. ILO tarafından 2021 yılında hazırlanan rapora göre, dünyada ve Türkiye’de ev eksenli çalışanların çoğunluğu toplumsal cinsiyete dayalı bir üretim biçimi olarak kadınlardan oluşmaktadır. Genelde eğitim düzeyi düşük ve vasıfsız kadınların ev ortamlarında çalışarak aile bütçelerine katkıda bulunmaları bu çalışma biçiminin artısını oluştururken, yapılan parça başı iş için ödenen ücretlerin düşük olması ve kadınların sigortasız ve sosyal güvencesiz dolayısıyla kayıt dışı olarak çalışmaları ev eksenli çalışma biçiminin olumsuz yönünü oluşturmaktadır. 5. Türkiye’de Kadınların Kayıt Dışı İstihdamı Türkiye benzer kültürel özellikler gösterdiği Müslüman ülkelerle kıyaslandığında, kadınların istihdam oranının daha yüksek olduğu ve kayıt dışı istihdamının da daha düşük olduğu görülmektedir. Ayla Yazıcı \| 137 137 Ayla Yazıcı Şekil 5: İşgücüne Dahil Olmayan Kadınların İş Aramama Nedenleri (bin kişi) Kaynak: TÜİK, İşgücü İstatistikleri 2022. Kaynak: TÜİK, İşgücü İstatistikleri 2022. TÜİK tarafından yayınlanan 2022 yılına ait işgücü istatistiklerine göre işgücüne dahil olmayan kadınların iş aramama nedenleri olarak en belirgin neden “ev işleriyle meşgul” olarak belirlenmiş. İşgücüne dahil olmayanların yaklaşık yarısını (10 milyonu) oluşturan bu kadınlar ücretsiz hane içi emek olarak adlandırılan ev işleri ve bakım hizmetini gerçekleştirmektedir. Aslında bu, kadınların toplum içerisinde ana görevinin ücret karşılığı bir işte çalışmalarından ziyade, bakım hizmetinde bulunmaları gerektiği düşüncesinin bir sonucudur (Bayar, 2021: 9). TÜİK tarafından yapılan aile yapısı araştırması sonuçlarına göre 2021 yılında kadınların çalışması ile ilgili bireylerin algıları incelendiğinde, kadınların büyük bir oranı için kadınların çalışması ve sosyal hayata katkı sağlaması değerli olarak görüldüğü belirtilse de, kadının asli görevinin çocuk bakımı ve ev işleri olduğu (%35,8), çalışan kadının evle ilgili sorumluluklarını ihmal edeceği (%27,7), aile geçimi için zorunlu değilse kadının çalışmaması gerektiği (%49,9) ve kadının çalışmasının geleneklerimize aykırı olduğunu 138 Türkiye’de Kadınların Kayıt Dışı İstihdamı (%13,9) belirten kadınların olması düşündürücüdür. Kadınların dahi kendilerine toplum tarafından biçilen ev kadını olup hane halkına bakmak ve ev işleri ile uğraşmak kalıbını gönüllü olarak benimsediklerini de göstermektedir. Şekil 6: Kadının Çalışması ile İlgili Algılar (2021 yılı, %) Şekil 6: Kadının Çalışması ile İlgili Algılar (2021 yılı, %) Kaynak: TÜİK, İstatistiklerle Kadın, 2022. Kaynak: TÜİK, İstatistiklerle Kadın, 2022. 6. Sonuç Son yıllarda Türkiye’de kayıt dışı istihdam oranları hem erkek hem kadınlar için düşüş eğilimine girmiş olsa da kayıt dışı istihdam ülkenin iş piyasasının önemli bir sorunu olmaya devam etmektedir. Düşük üretkenlik ve yoksullukla da ilişkilendiren kayıt dışılık, bireylerin ve işletmelerin sermayeye, krediye, teknolojiye, piyasalara ve resmi kurumlara erişimde daha büyük zorluklarla karşı karşıya kalmalarına neden olmaktadır. Dolayısıyla bu durum, bireyler açısından daha kötü çalışma koşulları, iş güvencesizliği, devlet yardımlarına ve sosyal güvenliğe erişim eksikliğine neden olurken, hükümetlerin de daha düşük vergi geliri elde etmelerine neden olmaktadır. Kayıt dışı istihdam hem erkekler hem de kadınlar açısından büyük bir öneme sahiptir, ancak özellikle Türkiye gibi gelişmekte olan ülkelerde 139 Ayla Yazıcı Ayla Yazıcı kadınların kayıt dışı istihdamı daha yüksek oranlarda gerçekleşmektedir. Kayıt dışı çalışan kadınlar ya daha düşük ücret almakta veya hiç ücret almayan, katkıda bulunan aile işçisi olarak faaliyet göstermektedirler. Türkiye nüfusunun yaklaşık yarısını oluşturan kadınlara ait istihdam ve işgücüne katılma oranlarının gelişmiş ülkelere kıyasla düşük olmasının nedeni genellikle kültürel ve sosyal normlara dayanmaktadır. Toplumda kadına evlenmek ve aile bakımını üstlenmek gibi temel bir rol biçildiğinden, günümüzde halen Türkiye’nin bazı bölgelerinde kızların gerekli eğitimden yoksun bırakılmaları ve dolayısıyla meslek edinerek aktif bir şekilde işgücü piyasasında resmi olarak görev almalarının önünde bir engel oluşturmaktadır. Ancak diğer taraftan Türkiye’de yaşanan yüksek enflasyon, gelir dağılımında adaletsizlik, yoksulluk gibi faktörler ise kadınları kayıt dışında çalışmaya itmektedir. 140 Türkiye’de Kadınların Kayıt Dışı İstihdamı 142 \| Türkiye’de Kadınların Kayıt Dışı İstihdamı 1 Doç.Dr. Çukurova Üniversitesi, Kozan İşletme Fakültesi, Adana, Türkiye, bgakar@cu.edu.tr, Orcid no: 0000-0001-7258-4402 Kaynakça Bayar, A. A. (2021). İşgücü Piyasasında Kadınların Yeri ve Politika Önerileri, İstanbul Politik Araştırmalar Enstitüsü (ISTANPOL). https://istanpol. org/post-i-sguc-piyasasinda-kad-nlar-n-yeri-ve-politika-onerileri (erişim tarihi: 20.11.2023) Chen, M. et.al. (2005). Progress of the World’s Women 2005: Women, Work and Poverty, United Nations Development Fund for Women (UNI- FEM). New York. unwomen.org/PoWW2005_eng.pdf (erişim tarihi: 21.11.2023) Chen, M. (2012). The Informal Economy: Definitions, Theories and Policies, Women in Informal Employment Globalizing and Organizing (WIEGO) Working Paper (1). Cambridge, MA, USA: WIEGO. http://wiego.org/ sites/wiego.org/files/publications/files/Chen_WIEGO_WP1.pdf (erişim tarihi: 21.11.2023) Chen, M. (2016). Expanding the Economic Potential of Women Informal Wor- kers, A Background Paper for the UN Secretary-General’s High-Level Pa- nel on Women’s Economic Empowerment WIEGO. expanding-econ-po- tential-informal-workers.pdf (erişim tarihi: 21.11.2023) Erdinç, Z. (2016). Türkiye’de Kayıt Dışı Ekonomiyle Mücadeleye Yönelik İzle- nen Politikalar ve Çözüm Önerileri, Anadolu Üniversitesi Sosyal Bilimler Dergisi, 16(4). 39-52. ILO (2018). Women and Men in the Informal Economy: A statistical picture. Geneva: ILO Publications. ILO (2021). Working from Home: From Invisibility to Decent Work, Geneva: ILO Publications. İstanbul Kalkınma Ajansı (2015). Kadın İstihdamının Artırılması Mali Destek Programı. Kul Parlak N, Çiftçi A.N. (2022). Pandeminin Kayıt Dışı İstihdamı Dışlama Et- kisi: Türkiye’de Formel-Enformel Emek Piyasalarındaki Ayrışma, Journal of Social Policy Conferences (82):93-135. The Seventeenth International Conference of Labour Statisticians (ICLS) (2003). Guidelines Concerning a Statistical Definition of Informel Employment. https://ilo.org/global/statistics-and-databases/standar- ds-and-guidelines/guidelines-adopted-by-international-conferences-of-la- bour-statisticians/WCMS_087622/lang--en/index.htm (erişim tarihi: 22.11.2023) The Seventeenth International Conference of Labour Statisticians (ICLS) (2003). Report of the Conference, International Labour Office, Geneva. www.ilo.org (erişim tarihi: 21.11.2023) 141 Ayla Yazıcı Shady, R. S. A. (2018). Echoing the Passive Voice: Women in Informal Small Business Sector: Challenges and Solutions, The Public Policy HUB, School of Global Affairs and Public Policy, AUC. Şahin, K. ve Develi, A. (2021). Kadın İstihdamında Kayıt Dışılık Sorunu. Kesit Akademi Dergisi, 7 (27), 386- 400. Yanıcı Erdal, İ. Ö. (2019). Türkiye’de Kayıt Dışı İstihdam ve Kayıt Dışı İstih- damla Mücadele Politikaları, Ufuk Üniversitesi Sosyal Bilimler Enstitüsü Dergisi (8)16. 225-246. Tiryaki-Yenilmez, D. (2023). Türkiye’de Kadınların Kayıt dışı İstihdamı Üzerine Kısa Bir Değerlendirme, Uluslararası Sosyal Hizmet Araştırmaları Der- gisi, 3 (2), 137-147. 142 \| Türkiye’de Kadınların Kayıt Dışı İstihdamı Bölüm 9 Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü Başak Gül Akar1 Özet Yapay zekâ ve evrensel temel gelir tartışmalarının önemli bir ilgi odağı haline geldiği günümüzde, yapay zekâ teknolojisi hızla geliştikçe ve istihdamı sekteye uğratma olasılığı arttıkça, iş kayıpları ve genişleyen gelir uçurumları konusundaki endişeler de yoğunlaşmaktadır. Mevcut tartışmalar, yapay zekâ ve otomasyonun iş piyasalarını eşit olmayan bir şekilde etkileyebileceğini ve çeşitli beceri seviyelerinde iş kayıplarına neden olabileceğini göstermektedir. Yapay zekâ ve otomasyonun olumsuz istihdam sonuçlarını hafifletmek için önerilen önde gelen çözümlerden biri evrensel temel gelir kavramıdır. Bu, kazançlarına veya servet durumlarına bakılmaksızın her yetişkine tutarlı ve koşulsuz bir miktar para sunulmasını gerektirmektedir. Yapay zekâ ve temel gelirle ilgili tartışmalar, teknolojik ilerlemenin iş kesintileri ve gelir eşitsizliği üzerindeki potansiyel olumsuz etkilerini azaltmaya odaklanmıştır. Evrensel bir temel gelir ve kazanılmış gelir vergisi kredileri gibi çözümler olası çareler olarak ortaya atılmıştır. Bu arada, yapay zekâ ve temel gelir konusundaki söylem, farklı bakış açıları ve çözüm önerileri sunan birden fazla tarafın dahil olduğu karmaşık bir yapıya sahiptir. Yapay zekâ ve otomasyonun yeni dijital ekonomi işleri vaat etmesine rağmen, işlerin önemli ölçüde yerinden edilmesinin önemli bir endişe kaynağı olmaya devam ettiği iddia edilmektedir. Yapay zekânın faydaları konusunda küçük bir grup yüksek vasıflı işçi öne çıkarken, diğerlerinin geride kalması nedeniyle de artan eşitsizlik potansiyeli yapay zekânın istihdam üzerindeki eksileri olarak değerlendirilmektedir. Önümüzdeki on yıllar için yapılan projeksiyonlar ve çalışmalar, robotların çeşitli ekonomik sektörlerde insan işçilerin yerini alabileceğini gösterse de henüz kesin cümleler kurmak için erken bir dönemde olduğumuzu gösteren çalışmalar da bir o kadar çoktur. Bununla birlikte bir önlem olarak, beklenen https://doi.org/10.58830/ozgur.pub401.c1757 https://doi.org/10.58830/ozgur.pub401.c1757 143 144 Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü bu değişim ve yaygın otomasyon karşısında hükümetlerin olası artan refah destekleri ile bunların finansmanı konusunda yaşanabilecek dengesizlikler nedeniyle bu sistemler üzerindeki yükün arttığı zorlayıcı bir görünüm söz konusu olabilecektir. Mevcut çalışma, evrensel temel geliri faydalı beklentilerini kabul ederek potansiyel bir çare olarak kabul etmektedir. Bununla birlikte, evrensel temel gelirin yaratabileceği olumsuz etkiler göz önüne alındığında, eğitim ve istihdam girişimlerine öncelik veren politikalara olan ihtiyacı da vurgulamaktadır. Giriş Bilgi teknolojilerinin ve bilişsel iş piyasasının yükselişi, yüksek vasıflı işgücü için ücretlerin artmasına neden olurken, düşük vasıflı işgücü otomasyon ve yapay zekâdan oldukça etkilenmektedir. Birçok çalışma ve uzman, yapay zekânın istihdamı yeniden şekillendirme ve dijital ekonomide yeni iş ve istihdam biçimleri yaratma potansiyeline dikkat çekmektedir (Santhi ve Muthuswamy, 2023; Kanazawa vd., 2022). Frey ve Osborne (2017) tarafından otomasyon ve bilgisayarlaştırmanın iş gücüne olan etkilerini değerlendiren önemli bir araştırma yapılmıştır. Araştırmacılar, ABD’deki işlerin yüzde kaçının otomasyona duyarlı olduğunu belirlemek için inceledikleri mesleklerin belirli özelliklerini (örneğin, manuel beceri gereklilikleri, sosyal zekâ gibi) değerlendirerek otomasyon riskini tahmin etmişlerdir. Çalışmada yapılan tahminlere göre, toplam ABD istihdamının yaklaşık %47’si yüksek risk kategorisinde yer almaktadır. Yani nispeten yakın zamanda, belki de önümüzdeki on veya iki yıl içinde otomatikleştirilebileceği düşünülen işler. Çalışmada ele alınan modele göre, taşımacılık ve lojistik mesleklerinde çalışanların çoğu, ofis ve idari destek çalışanlarının ve üretim mesleklerindeki işgücü risk altındadır. Bu bulgular literatürde belgelenen son teknolojik gelişmelerle tutarlıdır. Daha da şaşırtıcı olarak, hizmet mesleklerindeki istihdamın önemli bir kısmının bilgisayarlaşmaya karşı oldukça hassas olduğu tespit edilmiştir. Benzer ve farklı bulguları ile Autor (2015)’e ait çalışma da sektörel bulguları nedeniyle önem arz etmektedir. Buna göre, tarım (en altta) ile profesyonel, teknik ve yönetim (en üstteki üç grup) arasındaki geniş orta grupta ise hizmet ve vasıflı mavi yakalı meslekler sabit kalmış, büro/satış meslekleri yükselmiş, operatörlük ve işçilik meslekleri ise keskin bir düşüş göstermiştir. Hem yüksek hem de düşük eğitimli işlerde yaşanan hızlı istihdam artışı, “orta vasıflı” işlerin istihdamdaki payını önemli ölçüde azaltmıştır. Ancak Autor (2015), çalışmasını ılımlı bir şekilde sonlandırmakta ve önceki çalışmasından daha iyimser bir yaklaşım sunmaktadır. İstihdam kutuplaşmasının süresiz olarak (Autor, 2013 içinde Autor, 2015) devam etmeyeceği yönünde görüşle çalışmasını sonlandıran yazar, mevcut birçok orta beceri gerektiren işteki görevlerden 145 Başak Gül Akar \| Başak Gül Akar \| Başak Gül Akar bazıları otomasyona yatkın olsa da, birçok orta beceri gerektiren işin, beceri yelpazesinin tamamından bir görev karması talep etmeye devam edeceğini öngörmektedir. Örneğin, tıbbi destek meslekleri -radyoloji teknisyenleri, flebotomistler, hemşire teknisyenleri ve diğerleri- nispeten iyi ücretli, orta beceri gerektiren istihdamın önemli ve hızla büyüyen bir kategorisi olarak ortaya çıkmaktadır. Önümüzdeki on yıllarda, belirli mesleki becerileri temel orta beceri düzeyindeki okuryazarlık, sayısal okuryazarlık, uyum sağlama, problem çözme ve sağduyu ile birleştiren önemli bir orta beceri gerektiren iş tabakasının devam edeceği düşünülmektedir. Giriş Bu anlamda gelişmekte olan ülkelerde otomasyonun etkilerinin gelişmiş ülkelere kıyasla daha olumsuz olabileceği, zira işgücünü ikame eden yenilikler nedeniyle kaybedilebilecek daha fazla manuel iş bulunduğu ifade edilmektedir. Bu noktada evrensel temel gelir veya kazanılmış gelir kredisi gibi politikaların uygulanması, teknolojik değişim nedeniyle iş kaybı yaşayan bireyleri desteklemenin ve gelir eşitsizliğini ele almanın bir yolu olarak önerilmiştir (Papaioannou, 2021; Lu ve Zhou, 2021). Emek piyasasındaki bu gelişmelere karşı koymak için Bruun ve Duka (2018)’ya ait çalışmada, bir hükümetin işgücü piyasasındaki değişen manzarayı başarılı bir şekilde yönlendirmesine yardımcı olacak entegre iki yönlü çözüm önerilmektedir. Daha somut olarak Koşulsuz Evrensel Temel Gelir’in uygulamaya konulmasının öneminde söz eden çalışmada- devletten toplumdaki her vatandaşa koşulsuz, periyodik bir para transferi konu edilmektedir. Böyle bir gelir güvenliği yastığı altında, bir birey değişen ekonomiye uyum sağlama fırsatı verilirken teknolojik ilerlemenin olumsuz yönlerinden de korunmuş olacaktır. İnsanların hala makinelerle yarışabilmesini ve işgücü piyasasında varlıklarını sürdürebilmesini sağlamayı amaçlayan önerinin ikinci kısmını ise özellikle istihdam dışına itilenler için yeniden eğitim programları ve tüm eğitim sisteminin yapay zekâ çağıyla ilgili yaşam boyu, beceri temelli eğitime doğru yeniden yönlendirilmesi oluşturmaktadır. (Bruun and Duka, 2018). Mevcut çalışma giriş kısmında ana hatları ile yapay zekânın eşlik ettiği otomasyon, temel gelir ve istihdam ilişkisini aktardıktan sonra, günümüz çerçevesinde tartışmaların geldiği noktayı değerlendirmekte ve yapay zekânın istihdam üzerindeki olumlu ve olumsuz etkilerini sektörleri de dahil ederek zengin bir literatür çerçevesinde analiz edip, temel gelirin istihdam için bir çözüm olup-olmadığı noktasında sorgulamalar yapmakta ve sonuç kısmını politika önerileri ile genişleterek son bulmaktadır. 146 Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 1. Yapay Zekâ, İstihdam ve Temel Gelir Tartışmalarına Güncel Bir Bakış Yapay zekânın istihdam ve ekonomik yapılar üzerindeki etkisi, kapsamlı bir tartışma ve analiz konusu olmuştur. Yapay zekâ teknolojisinin gelişen manzarası, yalnızca potansiyel iş değiştirme ve gelir eşitsizliği ile ilgili tartışmaları beraberinde getirmekle kalmamış, aynı zamanda daha geniş sosyo-ekonomik yansımalar hakkında da düşüncelere yol açmıştır. (Gallego ve Kurer, 2022; Agrawal ve ark., 2019; Rydzik ve Kissoon, 2021; Ge ve ark., 2022; Bühler ve ark., 2022). Bazı çalışmalar, yapay zekânın önceki teknolojilerden farklı olabilecek dağılımsal sonuçlarla istihdamı önemli ölçüde yeniden şekillendirme potansiyeline sahip olduğunu öne sürmektedir. Örneğin, otomasyonun hem beceri artırıcı hem de eşitsizliği artırıcı olduğu düşünülmektedir. Son yıllarda, akademik araştırma odağında yapay zekânın endüstriyel iyileştirme ve istihdam uyumu üzerindeki etkisine doğru bir kayma olmuştur. Yapay zekâ, imalatın dönüşümüne ve iyileştirilmesine katkıda bulunmuştur, ancak aynı zamanda işgücü piyasası üzerinde de bazı etkilere yol açmıştır. Giriş Tekrarlayan görevleri içeren düşük vasıflı işgücü, yapay zekâ ile daha kolay ikame edilmekte, bu da hem düşük vasıflı hem de yüksek vasıflı işçiler için faydalarının eşitsiz dağılımı konusunda endişelere yol açmaktadır. Ayrıca çalışmalar, yapay zekânın istihdam kalıplarını, günlük yaşamları ve refahı etkileyerek toplumda temel bir yeniden yapılanma yaratma potansiyelini vurgulamaktadır (Kanazawa ve ark., 2022; Xiong ve ark., 2020). Yapay zekâ gelişmeye ve ilerlemeye devam ettikçe, toplum için oluşturduğu potansiyel riskleri ve faydaları göz önünde bulundurmak çok önemlidir. Bu, yalnızca makineleşmeden kaynaklanabilecek potansiyel işten çıkarma ve gelir eşitsizliğini değil, aynı zamanda bu tür ilerlemelerin getirebileceği daha geniş sosyo-ekonomik sonuçları da içermektedir. Önümüzdeki 10 veya 20 yıl içinde yapay zekâ ve talep arasındaki etkileşimi anlamak için, ampirik araştırmacıların birkaç özel soruya cevap vermesi gerekecektir. İlk olarak, yapay zekâ ne ölçüde insanların yerini alacak ve bunun yerine ne ölçüde sadece insan yeteneklerini artıracaktır? Yani, otomasyon meslekleri ne ölçüde tamamen otomatikleştirecek ve bunun yerine, bir meslek tarafından gerçekleştirilen tüm görevleri değil, yalnızca bazılarını ne ölçüde otomatikleştirecektir. İnsanlar tamamen ikame edilirse, talep artık istihdamı etkilemez çünkü insanlar için herhangi bir talep yoktur. Geçmişte, kapsamlı verimlilik artışına rağmen, teknoloji neredeyse her zaman işi yalnızca kısmen otomatikleştirmiştir (Kanazawa ve ark., 2022; Bessen, 2018). 147 Başak Gül Akar \| 147 Başak Gül Akar \| Yapay zekâ, inovasyonu kolaylaştırmak, üretkenliği artırmak ve işgücü talebini artırmakla ilişkilendirilmiş ve çeşitli sektörlerde olumlu ekonomik sonuçlara katkıda bulunmuştur. Bununla birlikte, artan işsizlik oranı ve yapay zekâ teknolojilerinde kodlanmış sistemik önyargılar potansiyeli hakkında endişeler de vardır. Yapay zekânın gelişiminin, iş yaratma ve yıkımı dengeleme, mahremiyet ve özerkliği sağlama ve toplumsal riskleri en aza indirme konusunda karar verilmesi gereken kritik bir noktada olduğuna inanılmaktadır. Bu anlamda, Avrupa Parlamentosu’nun 2025 yılında uygulamaya koyacağı yasa tasarısı tartışmanın gündemine oturmuştur. Avrupa Parlamentosu milletvekilleri, Avrupa’da yapay zekânın güvenli olmasını, temel haklara ve demokrasiye saygı gösterilmesini ve işletmelerin gelişip büyümesini sağlayacak bir yasa tasarısı üzerinde Konsey ile siyasi bir anlaşmaya varmıştır. Bu düzenleme, temel hakların, demokrasinin, hukukun üstünlüğünün ve çevresel sürdürülebilirliğin yüksek riskli yapay zekâdan korunmasını sağlamayı, aynı zamanda inovasyonu artırmayı ve Avrupa’yı bu alanda bir lider haline getirmeyi amaçlamaktadır. Kurallar, potansiyel risklerine ve etki düzeyine bağlı olarak yapay zekâ için yükümlülükler getirmektedir. Yapay zekânın belirli uygulamalarının vatandaşların haklarına ve demokrasiye yönelik potansiyel tehdidini kabul eden ortak yasa koyucular, hassas özellikleri kullanan biyometrik kategorizasyon sistemleri (örn. Giriş Yani yeni teknolojilerin geliştirilmesi ve bakımı, bilgi işlem, veri analizi gibi alanlarda yeni iş fırsatları oluşturabilecektir. Özellikle, manuel beceri gerektirmeyen, tekrar eden ve belirli algoritmalar tarafından yürütülebilecek işlerin otomasyona daha duyarlı olduğu belirlenmiştir. Düşük vasıflı ve tekrar eden işleri içeren meslek gruplarının otomasyon riski daha yüksek olarak değerlendirilmiştir. Örneğin, düşük vasıflı üretim işçileri ve rutin tabanlı ofis işleri, otomasyonun etkilerine daha fazla maruz kalan meslek grupları arasındadır. Buna mukabil, yaratıcılık, sosyal zekâ ve kompleks karar alma gerektiren meslek gruplarının otomasyona karşı daha dirençli olduğu bulgusu ortaya çıkmıştır. Yüksek vasıflı ve insan etkileşimi gerektiren işlerin otomasyon riski daha düşüktür. Bulgular, meslekler arasındaki otomasyon riskinin büyük bir değişkenlik gösterdiğini ve otomasyonun sadece belirli sektörleri etkileyeceğini göstermiştir. Bu, belirli beceri ve yeteneklere dayalı olarak otomasyon riskinin farklılık gösterdiği anlamına gelmektedir (Frey ve Osbourne, 2017). Bu noktada Bessen (2018), yapay zekâ teknolojilerinin birçok işi otomatikleştireceğini, ancak istihdam üzerindeki etkisinin açık olmadığını ima etmektedir. Sektörel olarak bakıldığında, imalat sektöründe teknolojinin, son yıllarda istihdamı keskin bir şekilde azalttığı görülmektedir. Ancak bundan önce, yüzyılı aşkın bir süre boyunca, hızlı teknolojik değişim yaşayan sektörlerde bile istihdam arttığına göre neyin değiştiği irdelenmelidir. Talep başlarda oldukça esnekken, daha sonra inelastik hale gelmiştir. Yapay zekânın istihdam üzerindeki etkisi de benzer şekilde talebin niteliğine bağlı olacaktır. Acemoglu ve Restrepo (2017), robotların Amerika Birleşik Devletleri (ABD)’ndeki iş piyasaları üzerindeki etkilerini inceledikleri çalışmada, robotlar ve otomasyonun ABD’de bazı işlerin kaybına yol açtığı ancak aynı zamanda yeni teknolojilere adaptasyon ve endüstriyel mekanizmaların etkileşimi çerçevesinde yeni iş yaratma potansiyelini de barındırdığı sonucuna varılmaktadır. Araştırma, robotların yerel iş piyasası üzerindeki etkilerini değerlendirirken toplam istihdam üzerinde 1990-2007 yılları arasında negatif etki olduğunu bulmuştur. Bununla birlikte, çalışma, teknolojik değişimin ve robotların kullanımı arttıkça bazı işlerin otomatikleştirilmesinin yanı sıra yeni iş tiplerinin ve iş alanlarının da ortaya çıktığını belirtmektedir. Bu durum, iş gücü piyasasına iki yönlü bir etki yaratmaktadır. Bunlarda ilki, iş kaybıdır. Buna göre, robot kullanımı arttıkça özellikle düşük ve orta beceri gerektiren işlerde çalışanların işsiz kalma riski artmaktadır. Diğeri, iş yaratımıdır. Yani yeni teknolojilerin geliştirilmesi ve bakımı, bilgi işlem, veri analizi gibi alanlarda yeni iş fırsatları oluşturabilecektir. Özellikle, manuel beceri gerektirmeyen, tekrar eden ve belirli algoritmalar tarafından yürütülebilecek işlerin otomasyona daha duyarlı olduğu belirlenmiştir. Düşük vasıflı ve tekrar eden işleri içeren meslek gruplarının otomasyon riski daha yüksek olarak değerlendirilmiştir. Örneğin, düşük vasıflı üretim işçileri ve rutin tabanlı ofis işleri, otomasyonun etkilerine daha fazla maruz kalan meslek grupları arasındadır. Giriş siyasi, dini, felsefi inançlar, cinsel yönelim, ırk); yüz tanıma veri tabanları oluşturmak için internetten veya kapalı devre televizyon (Close Circuit TeleVision-CCTV) görüntülerinden yüz görüntülerinin hedeflenmemiş bir şekilde kopyalanması; işyerinde ve eğitim kurumlarında duygu tanıma; sosyal davranışa veya kişisel özelliklere dayalı sosyal puanlama; özgür iradelerini atlatmak için insan davranışını manipüle eden otomasyon sistemleri; insanların savunmasızlıklarını (yaşları, engellilikleri, sosyal veya ekonomik durumları nedeniyle) istismar etmek için kullanılan yapay zekâ uygulamalarının yasaklanması yönünde kararlar alınmaktadır (Tran ve Nguyen, 2021; EP, 2023). Bununla birlikte, etik ve güvenlik kısmı ne kadar önem arz etse de robotların ve bilgisayarların yalnızca bir dizi rutin fiziksel iş faaliyetini insanlardan daha iyi ve daha ucuza gerçekleştirebildiği değil, aynı zamanda bilişsel yetenekler içeren faaliyetleri de giderek daha fazla gerçekleştirebildiği yeni bir otomasyon çağında yaşadığımızı kabul etmeliyiz. Bunlar arasında zımni yargılarda bulunma, duyguları algılama ve hatta araba kullanma gibi eskiden başarılı bir şekilde otomatikleştirmenin çok zor olduğu düşünülen faaliyetler yer almaktadır. Faaliyetlerin otomasyonu, hem bireysel süreç ve işletmeler düzeyinde hem de özellikle birçok ülkede çalışma çağındaki nüfusun payı azaldıkça üretkenliğin hızlanmasına şiddetle ihtiyaç duyulan Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 148 tüm ekonomiler düzeyinde üretkenlik artışı ve diğer faydaları sağlayabilecektir (Manyika ve ark., 2017). Bu noktada Bessen (2018), yapay zekâ teknolojilerinin birçok işi otomatikleştireceğini, ancak istihdam üzerindeki etkisinin açık olmadığını ima etmektedir. Sektörel olarak bakıldığında, imalat sektöründe teknolojinin, son yıllarda istihdamı keskin bir şekilde azalttığı görülmektedir. Ancak bundan önce, yüzyılı aşkın bir süre boyunca, hızlı teknolojik değişim yaşayan sektörlerde bile istihdam arttığına göre neyin değiştiği irdelenmelidir. Talep başlarda oldukça esnekken, daha sonra inelastik hale gelmiştir. Yapay zekânın istihdam üzerindeki etkisi de benzer şekilde talebin niteliğine bağlı olacaktır. Acemoglu ve Restrepo (2017), robotların Amerika Birleşik Devletleri (ABD)’ndeki iş piyasaları üzerindeki etkilerini inceledikleri çalışmada, robotlar ve otomasyonun ABD’de bazı işlerin kaybına yol açtığı ancak aynı zamanda yeni teknolojilere adaptasyon ve endüstriyel mekanizmaların etkileşimi çerçevesinde yeni iş yaratma potansiyelini de barındırdığı sonucuna varılmaktadır. Araştırma, robotların yerel iş piyasası üzerindeki etkilerini değerlendirirken toplam istihdam üzerinde 1990-2007 yılları arasında negatif etki olduğunu bulmuştur. Bununla birlikte, çalışma, teknolojik değişimin ve robotların kullanımı arttıkça bazı işlerin otomatikleştirilmesinin yanı sıra yeni iş tiplerinin ve iş alanlarının da ortaya çıktığını belirtmektedir. Bu durum, iş gücü piyasasına iki yönlü bir etki yaratmaktadır. Bunlarda ilki, iş kaybıdır. Buna göre, robot kullanımı arttıkça özellikle düşük ve orta beceri gerektiren işlerde çalışanların işsiz kalma riski artmaktadır. Diğeri, iş yaratımıdır. Giriş Buna mukabil, yaratıcılık, sosyal zekâ ve kompleks karar alma gerektiren meslek gruplarının otomasyona karşı daha dirençli olduğu bulgusu ortaya çıkmıştır. Yüksek vasıflı ve insan etkileşimi gerektiren işlerin otomasyon riski daha düşüktür. Bulgular, meslekler arasındaki otomasyon riskinin büyük bir değişkenlik gösterdiğini ve otomasyonun sadece belirli sektörleri etkileyeceğini göstermiştir. Bu, belirli beceri ve yeteneklere dayalı olarak otomasyon riskinin farklılık gösterdiği anlamına gelmektedir (Frey ve Osbourne, 2017). Gi i i (2023) diji l k l ji i ği i diğ l l d ülk l Giannini (2023), dijital teknolojinin, eğitim ve diğer alanlarda ülkeler içinde ve ülkeler arasında uçurumları genişletme konusunda rahatsız edici 149 Başak Gül Akar Başak Gül Akar bir geçmişe sahip olduğunu kabul etmekle birlikte, yapay zekâ teknolojisinin büyük olasılıkla çok sayıda işin otomasyonunu hızlandıracağını da gözden kaçırmamaktadır. Ayrıca, özellikle halihazırda yüksek ücretli alanlarda ve mesleklerde çalışan seçkin çalışanların üretkenliğini önemli ölçüde artırması muhtemeldir. Zaten birçok toplumda çok geniş olan eşitsizliğin yapay zekâ tarafından daha da genişlemesine direnmemiz gerekmektedir. Yeni teknoloji uygulamaları, eşitsizlik uçurumlarını kapatmayı ikinci bir düşünce olarak değil, bir başlangıç noktası olarak önceliklendirmelidir. Yapay zekânın işgücü üzerinde doğrudan bir azalma yaratmak yerine, genellikle mevcut iş rollerini değiştirdiği ve yeni iş rolleri yarattığı gözlemlenmiştir. Otomasyonun yarattığı zorlukların yanı sıra ekonomik ve sosyal fırsatlar da getirebileceği, iş gücü eğitiminin ve yeniden yetiştirilmenin önemini artırarak karşılanabilir. Özel bir vurgu yapmak gerekirse, ekonomik ve sosyal yapıları düzeltmek ve iş gücünü geleceğin işleri için hazırlamak adına politika yapıcılar, eğitim kurumları ve işletmeler tarafından atılacak adımlar büyük önem taşımaktadır. 2. Yapay Zekâ ve İstihdam İlişkisi Yapay zekâ ve otomasyondaki son gelişmeler, teknolojik ilerlemenin hızlı temposu ve yapay zekâ ile makine öğreniminin artan yetenekleri, istihdam üzerindeki etkinin geçmişte olduğundan daha şiddetli olabileceğine dair endişeleri artırmaktadır (Liu, Z., 2021). Manyika ve ark. (2017)’a ait çalışmada, bu endişelerin şiddeti konusunda biraz daha ılımlı bir yaklaşımın ortaya konulduğu görülmektedir. Buna göre, otomasyon bir gecede gerçekleşmeyecektir ve beş temel faktör benimsenme hızını ve kapsamını etkileyecektir. Birincisi teknik fizibilite, çünkü teknolojinin icat edilmesi, entegre edilmesi ve belirli faaliyetleri otomatikleştiren çözümlere uyarlanması gerekmektedir. İkincisi, benimseme için iş durumunu etkileyen çözüm geliştirme ve dağıtma maliyetidir. Üçüncüsü, otomasyona alternatif olarak insan emeğinin arzı, talebi ve maliyetleri de dahil olmak üzere işgücü piyasası dinamikleridir. Dördüncüsü, daha yüksek verim ve artan kalitenin yanı sıra işgücü maliyeti tasarruflarını da içerebilecek ekonomik faydalardır. Son olarak, mevzuat ve sosyal kabul, dağıtım iş açısından mantıklı olsa bile benimseme oranını etkileyebilir. Tüm bu faktörler dikkate alındığında, otomasyonun mevcut iş faaliyetleri üzerindeki etkisinin tam olarak ortaya çıkmasının on yıllar alacağını tahmin edilmektedir. Otomasyonun etkileri makro düzeyde tüm sektörlerde veya ekonomilerde yavaş olabilirken, mikro düzeyde, faaliyetleri otomatikleştirilen bireysel bir işçi veya sektörü otomasyon kullanan rakipler tarafından bozulan bir şirket için oldukça hızlı olabilir. 150 Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü Yapay zekânın istihdam üzerindeki etkisine ilişkin tartışmalar devam ederken, etkilenebilecek belirli sektörleri göz önünde bulundurmak önemlidir. Örneğin, imalat ve tarım gibi sektörler, otomatikleştirilmesi için yüksek teknik potansiyele sahip öngörülebilir fiziksel faaliyetler içermekte, ancak bazı gelişmekte olan ülkelerdeki düşük ücret oranları benimsemeyi kısıtlayabilmektedir. Birçok çalışan, çeşitli faaliyetler otomatikleştirilirken makinelerin yanında çalışmaya devam edecektir. Daha erken otomatikleşmesi muhtemel faaliyetler arasında, özellikle imalat ve perakende ticarette yaygın olan öngörülebilir fiziksel faaliyetlerin yanı sıra, tüm sektörler, beceriler ve ücretler yelpazesinde var olan faaliyetler olan veri toplama ve işleme yer almaktadır. Bazı otomasyon biçimleri beceri yanlı olacak, dosyalama memurları veya montaj hattı çalışanları gibi daha düşük becerili ve rutin yoğun mesleklere olan talebi azaltsa bile yüksek becerili çalışanların verimliliğini artırma eğiliminde olacaktır. Diğer otomasyonların orta vasıflı çalışanları orantısız bir şekilde etkilediği görülmektedir (Manyika ve ark., 2017). Sektörel etkileri biraz daha derinlemesine inceleyecek olursak, yapay zekâ ve diğer yeni nesil bilgi teknolojilerinin yüksek kaliteli gelişimi teşvik ettiği ve aynı zamanda robot üretim sürecini hızlandırdığı imalat sanayi dikkate değer bir önemi hak etmektedir. Bu hızlanma, sektördeki toplam istihdamı, istihdam yapısını ve istihdam kalitesini potansiyel olarak etkileyebilecektir. İmalat sanayinde yapay zekâ uygulaması, işin, mesleğin ve organizasyonun sınırlarını değiştirme potansiyeline sahiptir. 2. Yapay Zekâ ve İstihdam İlişkisi Çalışanların mesleki örüntüsünü etkileyebilmekte, bir kuruluş içindeki kaynakların düzenini ve dağılımını değiştirebilmekte, bireyler ve kuruluşlar arasındaki ilişkiyi bozabilmekte ve insan-organizasyon eşleşme modelini değiştirebilmektedir. Ek olarak, yapay zekânın istihdam üzerindeki etkisi imalat endüstrisinin ötesine uzanmaktadır. Sağlık ve yardım sektörleri de dahil olmak üzere çeşitli meslekleri kapsamaktadır. Geleneksel olarak otomasyona karşı bağışık olarak tasvir edilen sağlık ve yardım sektörlerindeki profesyonellerin istihdamı da yapay zekâdan etkilenebilir. Yapay zekâ ve otomasyondaki hızlı ilerlemeler, özellikle imalat ve sağlık gibi sektörlerde işlerin potansiyel olarak yer değiştirmesine ilişkin endişeleri artırmıştır. Ancak, yapay zekânın istihdam üzerindeki etkisinin niceliksel olmaktan çok niteliksel olacağını göz önünde bulundurmak önemlidir (Aroles ve ark., 2019; Innes ve ark., 2022). Konuya farklı bir çerçeveden yaklaşan Graham ve ark. (2017), online iş platformlarının gelişmekte olan ülkelerdeki insanlar için hem fırsatlar hem de riskler taşıdığını savunmaktadır. Çalışma, online iş platformlarının olumsuz etkilerini azaltmak için hükümetlerin ve şirketlerin önlem alması gerektiğini savunmaktadır. Çevrimiçi işlerde çalışmanın önemli ödülleri 151 Başak Gül Akar olsa da önemli riskleri de olduğunu belirtmek önemlidir. Ayrımcılık, düşük ücret oranları, fazla çalışma ve güvensizlik gibi risklerle doğrudan mücadele edilmesi gerekmektedir. Bu bağlamda, online iş platformlarının olumsuz etkilerini azaltmak için hükümetlerin ve şirketlerin önlem alması gereği vurgulanmaktadır. Graham ve ark. (2017), hükümetlerin online iş platformlarını düzenlemesi ve işçilerin haklarını koruması gerektiğini ve ayrıca, şirketlerin işçilere daha iyi ücretler ve çalışma koşulları sağlaması gerektiğini savunmaktadırlar. Frey ve ark. (2016), işlerin otomasyon nedeniyle ortadan kaldırılmasına karşı korunmak için, hangi özelliklerin belirli bir işin otomatikleştirilmesiyle ilişkili olma olasılığının en yüksek olduğunu bilmenin önemli olduğunu ifade etmekte ve algılama ve manipülasyon, yaratıcı zekâ ve sosyal zekânın otomasyonun önündeki darboğazlar olduklarını belirtmektedirler. İlgili çalışmaya göre, nitelikli sektörlere yatırım yapmış olan şehirler ve bölgeler otomasyona karşı nispeten güvende kalmaya devam edecek ve teknolojik dinamizm, istihdamı en üst düzeye çıkarmanın ve yeni teknolojilerden olumlu bir şekilde faydalanmanın en iyi yolu olmaya devam edecektir. Eğitim aynı zamanda politika yapıcıların hızlandırılmış teknolojik değişimin etkilerine hazırlanmak için yararlanmaları gereken çok önemli bir araçtır. Genel olarak, çalışma teknolojik gelişmelerin işgücü üzerinde önemli bir etkiye sahip olduğunu dile getirmekte ve önlem olarak eğitim ve istihdam politikalarını desteklemektedir (Frey ve ark., 2016). Benzer şekilde, Shook ve Knickrehm (2017)’a ait çalışmanın bulguları, çalışanların en iyi teknolojiden yararlanarak bu şekilde istihdam kaybı olmak yerine sayıyı yükselteceklerini ortaya koymaktadır. Elde edilen bulgulara göre, çalışanlar bu şekilde sadece iyi niyet göstermiyor, otomasyon karşısında ne yapması gerektiği konusunda da harekete geçiyorlar. Bu şekilde, yeni beceriler öğrenmeleri gerektiğini de anlıyorlar. 2. Yapay Zekâ ve İstihdam İlişkisi Bu anlamda, dijital öğrenmeyi, eğitimi günlük işe sorunsuz bir şekilde entegre ederek hızlanılabileceğini ifade etmektedirler. İlgili araştırmaya göre, otomasyon bekleyenlerin yüzde 80’i, otomasyonun önümüzdeki beş yıl içinde iş deneyimlerini nasıl etkileyeceği konusunda zorluklardan daha fazla fırsat öngörmektedir. Ayrıca, yapay zekânın tek başına yıllık ekonomik büyüme oranlarını ikiye katlama ve 2035 yılına kadar incelenen 12 gelişmiş ülkede işgücü üretkenliğini yüzde 40’a kadar artırma potansiyeline sahip olduğu bulunmuştur. ED (2023) yılına ait yapay zekâyla ilişkili olarak öğrenme ve öğretme üzerine olan rapor, son bulguları içeriyor olması nedeniyle eğitimin bu manadaki rolüne ilişkin katkı sunmaktadır. Rapor, yapay zekanın eğitim ve öğretimin geleceğinde önemli bir rol oynayacağını ve eğitimcilerin, politika yapıcıların ve araştırmacıların otomasyonun potansiyelini anlamaları ve yapay zekayı etkili bir şekilde kullanmaları 152 \| Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü fazla kullanılmaktadır ve dolayısıyla öğrenmeyi kişiselleştirmek, eğitim kalitesini artırmak ve öğretmenlerin iş yükünü azaltmak için kullanılabilir. Ancak yapay zekânın eğitim ve öğretimde kullanımı, etik endişeleri de beraberinde getirmektedir. Giannini (2023) de en son tartışmaları ele alması nedeniyle güncel bir bakış açısı sunduğu çalışmasında öğretmenlerden daha fazlasını talep ederken, yeni ve güçlü üretken yapay zekâ teknolojisinin, eski dijital araçlar ve hizmetlerle birlikte, öğretmenlerin otoritesini ve statüsünü zayıflatma potansiyeline karşı da dikkati çekmektedir. Ortaya çıkan yapay zekânın, yatırım seçimleri açısından da bir ikilem ortaya çıkardığının ifade edildiği çalışmada, “kamu yatırımları da dahil olmak üzere yatırımları, akıllı insanlar gibi davranan makinelerin yeteneklerini oluşturmaya mı yoksa yaşayan insanların yeteneklerini oluşturmaya mı yönlendirmeliyiz?” sorusu sorulmaktadır. Yakın geçmişte, “öğrenme”, “eğitim”, “koçluk”, “öğretim” gibi terimlerin insanlarla ilgili olduğundan emin olunabilecekken, bunun artık daha az netlik içerdiği ifade edilmektedir. Görüleceği üzere, yapay zekânın etkileri sektöre ve uygulama alanına göre değişkenlik göstermektedir. İstihdamın azalmasına yol açabilecek sektörlerin yanı sıra, yeni iş alanlarının ortaya çıkması ve mevcut işlerin içeriğinin değişmesi gibi etkileri de göz önünde bulundurmak önemlidir. Yapay zekânın etkisi, iş gücü piyasasının sadece mevcut iş pozisyonları ve sayısına odaklanmak yerine, işlerin niteliğine ve çalışma koşullarına nasıl bir değişiklik getirebileceği bağlamında değerlendirilmelidir. Bu dinamikler, kompleks ve çok boyutlu bir yaklaşım gerektirmektedir ve önümüzdeki dönemdeki politika oluşturma süreçlerinde ve eğitim sistemlerinin yeniden yapılandırılmasında anahtar rol oynayacaktır. Bu bağlamda, yapay zekânın iş gücü üzerindeki etkisinin yönetilmesi ve gelecek için işgücü stratejilerinin şekillendirilmesi sürecinde karar vericiler için becerilerin geliştirilmesi, esneklik ve uyum, yeni işlerin teşviki, yapay zekâ uygulamalarının toplumsal etkilerinin değerlendirilmesi gibi başlıkların gündemlerine taşınması önem arz etmektedir. 2. Yapay Zekâ ve İstihdam İlişkisi Yapay zekâ teknolojilerinin istihdam üzerindeki etkisinin anlaşılması ve bu değişim sürecinin yönetilmesi, hem hükümetlerin hem de özel sektörün işbirliğiyle yapılacak kapsamlı bir çabanın sonucunda mümkündür (WEF, 2023). 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Otomasyonla ilgili mevcut tartışmaların çoğu, insan emeği fazlalığına dayanan kitlesel işsizlik potansiyeline odaklanmış olsa da dünya ekonomisi aslında hem gelişmiş hem de gelişmekte olan ekonomilerdeki demografik yaşlanma eğilimlerinin üstesinden gelmek için robotlara ek olarak çalışan insan emeğine ihtiyaç duyacaktır. Bazı işler yapay zekâ ile değiştirilebilirken, farklı 153 Başak Gül Akar alanlarda yeni fırsatların da ortaya çıkacağını kabul etmek çok önemlidir. Bu nedenle, bireylerin gelişen iş piyasasında başarılı olmak için uyum sağlamaları ve gerekli becerileri edinmeleri çok önemlidir (Manyika ve ark., 2017). Bruun ve Duka (2018), yapay zekânın tüm iş yelpazesinde insan emeğini ikame etme potansiyeline sahip olduğunu, öyle ki hiç kimsenin mesleğinin savunulamazlığına inanmaması gerektiğini ifade etmektedir. Bu nedenle kararlarını ‘Şah Mat Senaryosu’na dayandırmaktadırlar, yani geleceğin makul, ancak kesin olmayan aşırı bir versiyonuna arkalarını yaslamaktadırlar. Ancak bu tahminin olasılığını çevreleyen belirsizliğin farkında olarak, yapay zekânın işgücü üzerindeki etkilerine ilişkin bir tartışmanın yine de gözden kaçırılmaması gerektiğine inanmaktadırlar. İlgili çalışmaya göre, Şah-Mat senaryosunun gerçekleşme olasılığı önümüzdeki 20 yıl içinde sadece %5 olsa bile, riskin kendisinin bu sonucu planlamak için yeterince zorlayıcı bir neden olduğuna dikkat çekmektedirler. Yapay zekâ, sürücüsüz otomobiller, otomatik çevrimiçi asistanlar ve sanal gerçeklik deneyimleri şeklinde günlük hayatımıza hızla giriyor. Bunu yaparken yapay zekâ, daha önce bilgisayarlaştırılamayacağı düşünülen alanlarda insan istihdamını ikame etmiş durumdadır. Mevcut eğilimlere dayanarak, emeğin teknolojik olarak yer değiştirmesinin gelecekte önemli olacağı tahmin edilmektedir ve eğer kontrol edilmezse, bu durumun felaket boyutlarında toplumsal işsizlik seviyelerine yol açabileceği varsayımı yapılmaktadır. Şu anda sergilenen teknolojiler göz önüne alındığında, çok az sayıda meslek - yüzde 5’ten az - bugün tam otomasyona adaydır, yani bu meslekleri oluşturan her faaliyet otomatikleştirilmiştir. Manyika ve ark. (2017), bu bulguyu destekler şekilde, faaliyetlerinin önemli bir yüzdesi otomatikleştirilebileceğinden, neredeyse her mesleğin kısmi otomasyon potansiyeline sahip olduğunu ifade etmektedirler. İlgili çalışmada, dünyadaki işgücünde insanların yapmak için ücret aldığı tüm faaliyetlerin yaklaşık yarısının, şu anda sergilenen teknolojilerin uyarlanmasıyla potansiyel olarak otomatikleştirilebileceği tahmin edilmektedir. Otomasyonun hızı ve kapsamı ve dolayısıyla çalışanlar üzerindeki etkisi, farklı faaliyetler, meslekler ve ücret ve beceri düzeyleri arasında değişiklik gösterecektir. Teknoloji gelişimi hem düşük vasıflı hem de yüksek vasıflı işçilerin faaliyetlerini otomasyona daha duyarlı hale getirdikçe, bu kutuplaşma etkileri azalabilecektir. Otomasyonun coğrafyalar ve sektörler arasında geniş çaplı etkileri olacaktır. Otomasyon küresel bir olgu olmasına rağmen, dört ekonomi -Çin, Hindistan, Japonya ve Amerika Birleşik Devletleri- toplam ücretlerin yarısından biraz fazlasını ve şu anda sergilenen teknolojilerin uyarlanmasıyla teknik olarak otomatikleştirilebilen faaliyetlerle ilişkili çalışan sayısının neredeyse üçte ikisini oluşturmaktadır. 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Ülkeler içinde otomasyon potansiyeli, sektör karışımından \| Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 154 otomasyona duyarlılık açısından sınıflandırıldığında özellikle, manuel beceri gerektirmeyen, tekrar eden ve belirli algoritmalar tarafından yürütülebilecek işlerin otomasyona daha duyarlı olduğu belirlenmiştir. Düşük vasıflı ve tekrar eden işleri içeren meslek gruplarının otomasyon riski daha yüksek olarak değerlendirilmiştir. Örneğin, düşük vasıflı üretim işçileri ve rutin tabanlı ofis işleri, otomasyonun etkilerine daha fazla maruz kalan meslek grupları arasındadır. Buna mukabil, yaratıcılık, sosyal zekâ ve kompleks karar alma gerektiren meslek gruplarının otomasyona karşı daha dirençli olduğu bulgusu ortaya çıkmıştır. Yüksek vasıflı ve insan etkileşimi gerektiren işlerin otomasyon riski daha düşüktür. Bulgular, meslekler arasındaki otomasyon riskinin büyük bir değişkenlik gösterdiğini ve otomasyonun sadece belirli sektörleri etkileyeceğini göstermiştir. Bu, belirli beceri ve yeteneklere dayalı olarak otomasyon riskinin farklılık gösterdiği anlamına gelmektedir (Frey ve Osborne, 2017). Bruun ve Duka (2018), okul müfredatı ve yeniden eğitim programlarındaki reformların eşlik ettiği bir Temel Gelir programının uygulamaya konulması yoluyla gelecekteki teknolojik işsizliği azaltmanın bir yolunu sunmaktadır. Böyle bir planın robotik işgücünden yararlanan endüstrilere uygulanacak özel bir vergi ile finanse edilebileceğini savunmakta ve bir hükümetin bu öneriyi kavramsal aşamadan alıp on yıl içinde ülke çapında uygulamasını sağlayacak pratik bir yol haritası çizmektedirler (Bruun and Duka, 2018). Temel gelir, yapay zekanın olumsuz etkilerini azaltmaya yardımcı olabilir. Yapılan araştırmalar, temel gelirin işsizlik oranlarını düşürebileceğini ve gelir eşitsizliğini azaltabileceğini göstermektedir. Ayrıca temel gelir, yapay zekanın olumlu etkilerini artırmaya da yardımcı olabilmektedir. Buna göre, temel gelir insanların yeni beceriler edinmelerini ve yeni işler kurmalarını kolaylaştırabilmektedir. Bu, yapay zekanın getirdiği yeni fırsatlardan yararlanılmasını kolaylaştırabilir ve ekonomik büyümeyi teşvik edebilir. Temel gelir, yapay zekanın etik ve sosyal etkilerini ele almaya da yardımcı olabilir. Bununla birlikte, temel gelirin yapay zekanın önyargılı olmamasını ve herkes için faydalı olmasını sağlayabileceği de diğer bulgular arasındadır. Genel olarak, yapay zekâ ve temel gelir arasındaki ilişki karmaşıktır ve birçok faktöre bağlıdır. Ancak, temel gelir, yapay zekanın olumsuz etkilerini azaltmaya ve olumlu etkilerini artırmaya yardımcı olabilir (Standing, 2017; WEF, 2023; Zittrain, 2008). Autor (2015)’un otomatizasyon ve gelirle ilgili bulguları, teknolojik ilerlemenin işgücü piyasasına etkilerinin çeşitli boyutlarına odaklanmaktadır. Genel olarak Autor (2015), otomosyonun gelir dağılımı üzerindeki etkilerini polarizasyon, ücret eşitsizliği, gelirin teknolojik ilerlemeye tepkisi doğrultusunda incelemektedir: Buna göre, iş piyasası yüksek ve düşük beceri 155 Başak Gül Akar gerektiren işler olarak farklılaşırken, orta beceri gerektiren işler otomatizasyon nedeniyle azalmaktadır. Yüksek becerili işler, daha yüksek gelir getirirken, düşük becerili işler genellikle düşük gelirle ilişkilendirilmektedir. 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Teknolojik değişiklikler, yüksek becerili çalışanlara olan talebi artırırken düşük ve orta becerili işlerin otomatize edilmesine neden olmuştur. Bu durum, yüksek becerili ve düşük becerili çalışanlar arasında bir ücret eşitsizliği meydana getirebilecektir. Son olarak, eğitimli ve teknoloji kullanmayı iyi bilen işçilerin, bu ilerlemelerden gelir anlamında daha çok yararlandığına dikkat çekilmektedir. Frey ve Osborne (2017), yapay zekanın, önümüzdeki 20 yıl içinde ABD’deki işlerin neredeyse yarısını otomasyona bağlayacağını ifade ederken, temel gelirin işsiz kalan kişilerin geçimlerini sağlamalarına yardımcı olabileceğini ve gelir eşitsizliğini azaltabileceğini de belirtmişlerdir. Temel gelir, insanların yeni beceriler edinmeleri ve yeni işler kurmaları için daha fazla zaman ve kaynak sağlayabilir. Standing (2017) destekler şekilde, temel gelirin politik ve ekonomik boyutlarını ve bunun insanların yaşamlarını nasıl iyileştirebileceğini tartıştığı eserinde, temel gelirin güvencesiz işlerde çalışan veya işsiz kalan kişilere ekonomik güvenlik sağlayarak toplumsal istikrarı artıracağını ve yaratıcı faaliyetlerle kişisel gelişim için zaman yaratacağını savunmaktadır. Susskind (2020), aynı şekilde teknolojik gelişmelere karşı nasıl cevap verebileceğimizi tartıştığı kitabında, teknolojik işsizlik ve otomasyon arttıkça insanların nasıl etkileneceğine odaklanmaktadır. Hem Standing (2017) hem de Susskind (2020), temel gelirin günümüzdeki sosyoekonomik problemlere çözüm olabileceğini ve yapay zekâ ile otomasyonun neden olduğu işsizlik ve gelir güvensizliğine karşı bir önlem teşkil edebileceğini vurgulamaktadır. Birçok ülkede temel gelirin yeni bir sosyal koruma politikası olarak benimsenmesine yönelik ilgi giderek artmaktadır. Güney Kore’nin de bu konuda bir istisna olmadığı vurgusu ile Kim ve Lee (2021), evrensel temel gelirin insanlığın genel refahı için bir yöntem olarak kullanılma potansiyelini inceledikleri çalışmada, temel gelirin çeşitli sosyoekonomik kontekstlerde uygulanabilirliğini ve etkililiğini belirlemeye yönelik bir Temel Gelir Sadakat İndeksi geliştirmiştir. Bu indeks, bir evrensel gelir programının özelliklerini ve başarısını ölçmek için kritik faktörleri tanımlamaktadır. Çalışmanın ana motivasyonlarından biri, Güney Kore’deki temel gelir tartışmalarında temel gelirin net bir tanımının yapılmamış olmasıdır. Temel gelir tartışmalarının yapılabileceği bir çerçeve geliştirmek için çok az çaba sarf edilmiştir. Temel gelirin yeni bir sosyal koruma politikası olarak benimsenmesi tartışmaları Güney Kore’de bilimsel bir referans noktası olmaksızın büyük ölçüde siyasi çıkarlara dayandırılmıştır. Temel geliri savunanlar ve eleştirenler, çıkarlarına bağlı olarak farklı isimler Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 156 kullandıklarından, temel gelir kavramında giderek artan bir kafa karışıklığı yaşanmaktadır. Temel gelirin tanımı ve özellikleri, temel gelirin geleneksel kamu yardımı, sosyal sigorta, sosyal ödenek, vergi kredisi ve acil durum yardımlarından farklı, kendine özgü bir nakdi yardım olduğunu açıkça göstermektedir. 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Temel gelir kendine özgü bir nakdi yardım olduğu için, eğer bir program temel gelirin tüm özelliklerine sahip değilse, temel gelir olarak etiketlense bile temel gelir değildir. Öte yandan, bu şekilde etiketlenmemiş bir program tüm özelliklere sahipse temel gelirdir. “Yeterlilik”, ödemenin alıcıların temel ihtiyaçlarını karşılayacak düzeyde olması gerektiği anlamına gelir. Diğer bir deyişle, eğer ödeme düzeyi temel ihtiyaçları karşılayamayacak kadar düşükse, temel gelir olarak adlandırılamaz. Kyun ve Joo (2020)’nun Temel Gelir Sadakat İndeksi gibi araçların yardımıyla, evrensel temel gelirin farklı toplumlar üzerindeki etkilerini özgün biçimde değerlendirme imkânı bulunmaktadır. Bu tür analitik araçlar, politika yapıcıların ve araştırmacıların, her bir ülkenin ihtiyaçlarına ve koşullarına uygun özelleştirilmiş yaklaşımlar geliştirmelerine yardımcı olabilecektir. Bu anlamda örnek teşkil edebilecek sosyal refah uygulamaları bağlamında hükümetlerin öncülük ettiği yoksullukla mücadele programları, dünya genelinde aşırı yoksulluğun ortadan kaldırılmasına yardımcı olma konusunda çok önemli bir role sahiptir. Ancak bu transferlerin biçimi ve özellikle de evrensel mi yoksa yoksullara yönelik mi olması gerektiği konusunda önemli sorular bulunmaktadır. Kamu tarafından finanse edilen ve kullanan ailelere ücretsiz olan ilkokullar gibi bazı hükümet programları evrensel olsa da günümüzde çoğu nakit transferi programı bir tür hedefleme mekanizması içermektedir. Hanna ve Olken (2018), gelirlerin nüfusun çok büyük bir kısmı için gözlemlenemediği gelişmekte olan ülkelerde bu değiş tokuş hakkında nasıl düşünülebileceğini araştırmışlardır. Bu ortamda, evrensel transferler gelir dağılımında yukarı çıkıldıkça kolayca geri vergilendirilemez. Sonuç olarak, evrensel transfer programları gelir dağılımının oldukça yukarısında aynı net transferi verecektir ve bu nedenle evrensel bir temel gelir programı ile diğer yollarla hedeflenen transferler arasındaki seçim, dışlama hatasını ortadan kaldırmak ile faydalanıcı başına çok daha küçük transferler vermek arasında çok önemli bir değiş tokuş anlamına gelmektedir. Endonezya ve Peru’dan elde edilen kanıtlar, gelişmekte olan ülkelerdeki mevcut hedefleme yöntemlerinin, kusurlu olmakla birlikte, evrensel programlara kıyasla refahta önemli iyileşmeler sağladığını göstermektedir, çünkü bu yöntemler evrensel programlara kıyasla yoksullara yararlanıcı başına çok daha fazla transfer yapabilmektedir. Bu programların birincil dezavantajı yatay eşitliktir - hedefleme kusurlu olduğu için, çatlaklardan kayan ve dışlanan önemli sayıda yoksul hane olacaktır. Bununla birlikte, birçok gelişmekte olan ülke - hedefleme kusurlu olduğu için, çatlaklardan kayan ve dışlanan önemli sayıda yoksul hane olacaktır. Bununla birlikte, birçok gelişmekte olan ülke 157 Başak Gül Akar Başak Gül Akar için simülasyonlar, hedeflemeden elde edilen refah kazançlarının önemli olabileceğini göstermektedir (Hanna ve Olken, 2018). Konunun biraz da magazinsel yönüne değinilecek olursa, güncel tartışmalara katılan Elon Musk (2017)’ın verdiği bir röportaj da kayda değer öneriler içermesi nedeniyle önemlidir. 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Tesla CEO’su Elon Musk (2017), verdiği bir röportajda, otomasyonun neden olacağı yaygın işsizliğe olası bir çözüm olarak evrensel temel geliri destekleyen teknoloji yöneticilerinin giderek artan listesine katılmıştır. Evrensel temel gelir, tüm vatandaşların gıda, kira ve kıyafet gibi temel harcamalarını karşılamak üzere her ay standart bir miktar para aldığı bir sistemdir. Musk (2017), ‘otomasyonla birlikte bolluk yaşanacağını ve neredeyse her şeyin çok ucuzlayacağını’ ifade ettiği konuşmasında, paranın teorik olarak insanlara çalışmasalar bile finansal güvence sağlamak için yeniden dağıtılabileceğinin üzerinde durmuştur. Hükümetin bir tür evrensel temel gelir ya da kaybedilen işleri telafi etmek için tüm vatandaşlara nakit para dağıtımı uygulaması gerekeceğini beklediğini söyleyen Musk (2017), otomasyon nedeniyle evrensel bir temel gelir ya da buna benzer bir şeyle karşılaşma ihtimalimizin oldukça yüksek olduğunu ifade etmiştir (Musk, 2017; Weller, 2017). Ancak burada temel gelirin nasıl uygulanacağı meselesi hassasiyet taşımaktadır. Standing (2017), temel gelirin kimlere, kimler tarafından ne şekilde ödenmesi noktasında pilot bir uygulama için şu adımları sıralamaktadır: i. Ödenen miktar temel olmalı, alıcı için anlamlı olacak kadar yeterli olmalı ancak tam bir güvenlik sağlayacak kadar yüksek olmamalıdır; ii. Gelir nakit olarak ya da örneğin bir banka hesabı, akıllı kart ya da cep telefonu aracılığıyla kolayca nakde çevrilebilecek bir biçimde ödenmelidir. Düzenli, öngörülebilir ve istikrarlı olmalı, tercihen sürekli bir dönem boyunca aylık olarak ödenmelidir. Esas olarak ‘irade zayıflığı’ etkisi nedeniyle toplu ödeme şeklinde yapılmamalıdır; iii. Temel gelir evrensel olmalı, başlangıçta genellikle pilot toplulukta ikamet eden herkese ödenmelidir. Bu, ekonomik olmayan etkiler de dahil olmak üzere toplumsal etkilerin tespit edilmesini sağlayacaktır. Ayrıca geri çekilemez olmalıdır; iv. Bu da hedefleme olmaması gerektiği anlamına gelmektedir. Temel gelir, yoksulluk nasıl tanımlanırsa tanımlansın, sadece ‘yoksul’ olduğu düşünülen kişilere verilmemelidir. Temel gelirin bir hak olması amaçlanmaktadır ve haklar evrenseldir; v. Aynı zamanda seçicilik de olmamalıdır. ‘Hak eden’ bir grup yerine başka bir gruba vermek, seçilen grubu diğerleriyle paylaşma baskısı altına sokarak temel gelirin etkisini azaltabilir. Sadece annelere gelir ödenmesi hane içinde gerilim yaratma riski taşır; vi. Temel gelir, önceden belirlenmiş bazı davranışlar gerektirmeden koşulsuz olmalıdır; vii. Temel gelir her erkeğe ve her kadına ayrı ayrı ve eşit olarak ödenmelidir. Temel gelir güvencesi bireysel bir haktır ve büyüklüğü ve Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 158 bileşimi politikanın kendisinden etkilenebilecek bir ‘aile’ veya ‘hane’ kavramına uygulanmamalıdır. Çocuklar ya da engellilik veya kırılganlık nedeniyle kendi gelirini toplayamayan yetişkinler için, onlar adına geliri almak üzere bir vekil (çocuklar söz konusu olduğunda anne) belirlenebilir; viii. 3. İstihdam Sorunu İçin Evrensel Gelir Bir Çözüm Mü? Etkilerinin adil bir şekilde değerlendirilmesini sağlamak için pilot uygulamanın başlangıcında veya pilot uygulama sırasında başka bir politika değişikliği yapılmamalıdır. Standing (2017), bu noktada pilot uygulamalara dair önemli bir hususu dile getirmeyi de ihmal etmemektedir. Bu tarz uygulamaların motivasyonu özgürlüğü, sosyal adaleti ve ekonomik güvenliği arttırırken, başka müdahalelere ihtiyaç duyabilecek davranışsal unsurları da test etmek olmalıdır. Pilot uygulamaların özgürlükten ödün vermesi, sosyal adalet ve güvenlik ilkelerini zedelemesi halinde bu uygulamalara karşı çıkılmalıdır. 4. Sonuç ve Politika Önerileri Teknolojik gelişmeler, işgücünde önemli değişikliklere yol açmıştır ve bu değişiklikler devam edecektir. Teknolojik gelişmelerin işgücü üzerindeki etkilerini azaltmak için eğitim ve istihdam politikaları önemlidir. Eğitim sistemleri, teknolojik gelişmelere ayak uydurmalı ve işçilere yeni beceriler kazandırmalıdır. Hükümetler, işsiz kalan kişilere destek sağlamalı ve yeni iş fırsatları yaratmalıdır. Şirketler, işçilere daha iyi ücretler ve çalışma koşulları sağlamalıdır (Frey ve ark., 2016). Shook ve Knickrehm (2017), liderlerin geleceğin işgücünü yönlendirmesine ve şekillendirmesine yardımcı olmak için şu önerilerde bulunmaktadır: Yeniden beceri edindirmenin hızlandırılması gereğine dikkat çekilen çalışmada, yukarıdan aşağıya, yaratıcılık ve yargı içeren teknik ve daha insani becerilere yatırım yapılması tavsiyesi ile birlikte, çalışanların yüzde 85’inin yeni beceriler öğrenmek için önümüzdeki altı ayda boş zamanlarını bu şekilde kullanmaları önerisi yapılmaktadır. Yeniden beceri edindirmeyi sağlarken dijital teknolojiden yararlanılması ve çalışanlar görevlerini yerine getirirken teknik tavsiye ve bilgi sağlayan akıllı gözlükler gibi giyilebilir teknolojilerin de buna dahil olabileceği ifade edilmektedir. Teknolojik gelişmeler, işgücünde önemli değişikliklere yol açmıştır ve bu değişiklikler devam edecektir. Teknolojik gelişmelerin işgücü üzerindeki etkilerini azaltmak için eğitim ve istihdam politikaları önemlidir. Eğitim sistemleri, teknolojik gelişmelere ayak uydurmalı ve işçilere yeni beceriler kazandırmalıdır. Hükümetler, işsiz kalan kişilere destek sağlamalı ve yeni iş fırsatları yaratmalıdır. Şirketler, işçilere daha iyi ücretler ve çalışma koşulları sağlamalıdır (Frey ve ark., 2016). Shook ve Knickrehm (2017), liderlerin geleceğin işgücünü yönlendirmesine ve şekillendirmesine yardımcı olmak için şu önerilerde bulunmaktadır: Yeniden beceri edindirmenin hızlandırılması gereğine dikkat çekilen çalışmada, yukarıdan aşağıya, yaratıcılık ve yargı içeren teknik ve daha insani becerilere yatırım yapılması tavsiyesi ile birlikte, çalışanların yüzde 85’inin yeni beceriler öğrenmek için önümüzdeki altı ayda boş zamanlarını bu şekilde kullanmaları önerisi yapılmaktadır. Yeniden beceri edindirmeyi sağlarken dijital teknolojiden yararlanılması ve çalışanlar görevlerini yerine getirirken teknik tavsiye ve bilgi sağlayan akıllı gözlükler gibi giyilebilir teknolojilerin de buna dahil olabileceği ifade edilmektedir. Ayrıca, bir bireyin yaşam boyu öğrenme ihtiyaçlarını desteklemek için öneriler sunan kişiselleştirilmiş eğitimler için akıllı yazılımların bu sürece dahil olabileceği tavsiyesinde bulunulmaktadır. Bunlara ilaveten, insan potansiyelini açığa çıkarmak için işin yeniden tasarlanması gereği üzerinde durulan çalışmada, çalışanların daha çeşitli iş ve esnek düzenlemeler taleplerini karşılamak için rol temelli, iş benzeri istihdam fırsatlarını birlikte yaratması ve en iyi yeteneklerin sadık kalmasını sağlayan cazip bir topluluk oluşturmak için hem çalışanlara hem de serbest çalışanlara bir dizi kaynak ve hizmetin sunulabileceği platformlar geliştirilmesinin önemi üzerinde durulmaktadır. Son olarak, yetenek havuzunun kaynağından güçlendirilmesi Ayrıca, bir bireyin yaşam boyu öğrenme ihtiyaçlarını desteklemek için öneriler sunan kişiselleştirilmiş eğitimler için akıllı yazılımların bu sürece dahil olabileceği tavsiyesinde bulunulmaktadır. 4. Sonuç ve Politika Önerileri Bunlara ilaveten, insan potansiyelini açığa çıkarmak için işin yeniden tasarlanması gereği üzerinde durulan çalışmada, çalışanların daha çeşitli iş ve esnek düzenlemeler taleplerini karşılamak için rol temelli, iş benzeri istihdam fırsatlarını birlikte yaratması ve en iyi yeteneklerin sadık kalmasını sağlayan cazip bir topluluk oluşturmak için hem çalışanlara hem de serbest çalışanlara bir dizi kaynak ve hizmetin sunulabileceği platformlar geliştirilmesinin önemi üzerinde durulmaktadır. Son olarak, yetenek havuzunun kaynağından güçlendirilmesi 159 Başak Gül Akar Başak Gül Akar ve bunun için toplu çözümlerin desteklenmesi gereği üzerinde durulmaktadır. Kamu-özel ortaklığı, sektörel ortaklıklar yanında eğitim sektörüyle birlikte çalışmanın avantajları aktarılmaktadır. ED (2023)’ın özellikle eğitim alanına yönelik tavsiyelerinde ise özellikle şu konuların altı çizilmektedir: Eğitimciler, yapay zekanın eğitim ve öğretimdeki potansiyelini anlamaları ve yapay zekayı etkili bir şekilde kullanmaları için eğitilmelidir. Okullar ve üniversiteler, otomasyon sistemlerini etik ve sorumlu bir şekilde kullanmaları için politikalar geliştirmelidir. Hükümetler, makineleşmenin eğitim ve öğretimde kullanılmasını desteklemek için yatırım yapmalı ve düzenlemeler geliştirmelidir. Yapay zekâ sistemleri, tarafsız ve adil olmalı ve öğrencilerin gizliliğini korumalıdır. Yapay zekâ, öğrencilerin eleştirel düşünme ve problem çözme becerilerini geliştirmelerine ve öğrencilerin yaratıcılıklarını ve yenilikçiliklerini geliştirmelerine yardımcı olmak için kullanılmalıdır. Başka bir deyişle, otomasyon yaygın bir şekilde kullanılmadığı sürece insan emeği fazlası oluşması, insan emeği açığı oluşmasından çok daha az olasıdır. Ancak işin doğası değişecektir. Süreçler bireysel faaliyetlerin otomasyonu ile dönüştürüldükçe, insanlar makinelerin yaptığı işi tamamlayıcı faaliyetler gerçekleştirecektir (ve bunun tersi de geçerlidir). Bu değişimler şirketlerin organizasyonunu, endüstrilerin yapısını ve rekabet temellerini ve iş modellerini değiştirecektir. İş dünyası için otomasyonun performans faydaları nispeten açıktır, ancak politika yapıcılar için konular daha karmaşıktır. Ekonomilerinin verimlilik artışı potansiyelinden faydalanma fırsatını benimsemeli ve sürekli ilerleme ve inovasyonu teşvik etmek için yatırım ve piyasa teşviklerini teşvik edecek politikaları uygulamaya koymalıdırlar. Aynı zamanda, çalışanların ve kurumların istihdam üzerindeki etkilere uyum sağlamalarına yardımcı olacak politikaları geliştirmeli ve yenilemelidirler. Bu, muhtemelen eğitim ve öğretim, gelir desteği ve güvenlik ağlarının yanı sıra yerinden edilenler için geçiş desteğinin yeniden düşünülmesini de içerecektir. İşyerindeki bireylerin günlük faaliyetlerinin bir parçası olarak makinelerle daha kapsamlı bir şekilde etkileşim kurmaları ve yeni otomasyon çağında talep görecek yeni beceriler edinmeleri gerekecektir. Otomasyon teknolojilerinin on yıllar boyunca işgücünde yaratabileceği değişimlerin ölçeği, 20. yüzyılda gelişmiş ülkelerin işgücünün tarımdan uzaklaşmasına neden olan uzun vadeli teknoloji destekli değişimlere benzer bir büyüklüktedir. Bu değişimler uzun vadeli kitlesel işsizlikle sonuçlanmak yerine o zamanlar öngörülmeyen yeni iş türlerinin yaratılmasına eşlik etmişlerdir (Manyika ve ark., 2017). 4. Sonuç ve Politika Önerileri Bu anlamda Deming (2015)’e ait, işgücü piyasasının sosyal becerileri giderek daha fazla ödüllendirdiği bulgusunun yer aldığı çalışmanın özellikle genç istihdamı oluşturan bireyler için yönlendirici olacağı düşünülmektedir. İ Yapay Zekanın İstihdam Alanındaki Güncel Etkileri ve Temel Gelirin Rolü 160 etkileşim gerektiren işler, ABD işgücünün payı olarak yaklaşık yüzde 12 puan artmıştır. Matematik yoğun ancak daha az sosyal işler - birçok STEM mesleği dahil - aynı dönemde yüzde 3,3 oranında küçülmüştür. İstihdam ve ücret artışı özellikle yüksek düzeyde hem matematik becerisi hem de sosyal beceri gerektiren işlerde güçlü olmuştur. İşgücü piyasasında sosyal becerilerin artan öneminin nedenlerinden biri, bilgisayarların insan etkileşimini taklit etme konusunda hala çok zayıf olmasıdır. Başkalarının zihnini okumak ve tepki vermek bilinçsiz bir süreçtir ve sosyal ortamlardaki beceri insanlarda binlerce yıl içinde gelişmiştir. İşyerindeki insan etkileşimi, çalışanların birbirlerinin güçlü yönlerinden yararlanarak ve değişen koşullara esnek bir şekilde uyum sağlayarak ekip halinde üretim yapmasını içermektedir. Bu tür rutin olmayan etkileşim, insanların makinelere karşı avantajının merkezinde yer almaktadır. Sosyal beceri yoğun meslekler için 1980 ve 2012 yılları arasında güçlü göreli istihdam ve ücret artışına dair kanıtlarla desteklenen çalışmada Deming (2015), yüksek düzeyde bilişsel beceri ve sosyal beceri gerektiren işlerin özellikle iyi performans gösterirken, yüksek matematik, düşük sosyal beceri gerektiren işlerin (birçok STEM mesleği dahil) özellikle kötü performans gösterdiğini ortaya koymaktadır. McAfee ve Brynjolfsson (2018), Deming (2015)’in ulaştığı sonuçları da değerlendirdikleri çalışmalarında, iş dünyasında sosyal görevlerin süregelen merkeziyetçiliğinin birbirine son derece bağlı üç ana nedeni olduğunu dile getirmektedirler. Bunlardan ilki ve en bariz olanı, dünyanın çok karmaşık ve hızla değişen bir yer olmasıdır. Bunun içinde gelişmek, büyük ölçüde sürekli koordinasyon gerektirir ve bunların hepsi otomatik güncellemeler ve sosyal medyadaki akranlar arasındaki konuşmalar yoluyla gerçekleştirilemez. Gerçekte iyi yöneticiler bir organizasyonun aktarımının sorunsuz bir şekilde işlemesini sağlarlar ve organizasyonun ele geçirilmesini önlerler. İnsanların sosyal becerilerinin bu kadar değerli olmasının ikinci nedeni, çoğumuzun sayıları ve algoritmaları tek başına ikna edici bulmamasıdır. İstatistiksel olarak anlamlı sonuçlarla dolu bir tablodan ziyade iyi bir hikâye veya ilgi çekici bir anekdottan çok daha fazla etkileniriz. Bu da bilişsel önyargılarımızdan bir diğeri, ancak hiçbirimizin görmezden gelemeyeceği bir durumdur. Dolayısıyla, akıllı şirketler sadece müşterilerini değil, kendi çalışanlarını da nazikçe ikna etme sanatına büyük yatırım yaparlar. Üçüncü neden en belirsiz olanıdır, ancak muhtemelen en önemlisidir. Biz insanlar birlikte çalışmak ve birbirimize yardım etmek isteriz ve bunu yapmaya teşvik edilebiliriz ve edilmeliyiz. 4. Sonuç ve Politika Önerileri b l l d k i l Bütün bu çalışmalar ışığında, yapay zeka ve otomasyonun iş gücüne olan etkisi artarken, devletler ve politika yapıcılar için temel gelir gibi yenilikçi politikaları araştırmak ve düşünmek daha da önem kazanmaktadır. Evrensel temel gelir, sosyal refah ve iktisadi esneklik arasında bir denge kurmayı vaat 161 Başak Gül Akar ederken, gelecek nesillere yönelik daha işlevsel ve kapsayıcı bir sosyal güvenlik ağı sunma potansiyeline sahiptir. Bu nedenle, yapay zeka bağlamında istihdam ve temel gelir konusundaki tartışmaların ve araştırmaların genişletilmesi, hem akademik çevrelerde hem de politika yapım süreçlerinde büyük bir değer taşımaktadır. 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Bölüm 10 Özet Özet Hükümetler, sosyal hizmetler, güvenlik ve alt yapı yatırımları gibi faaliyetlerinin devamı için gelire ihtiyaç duymaktadırlar. Vergiler, hükümetlerin faaliyetlerinin yürütülmesi için geliri artırmanın bir yoludur. Her ne kadar vergi gelirleri ekonomik büyümeyi teşvik edici ve finansman ihtiyacını karşılayan bir araç olarak kabul ediliyor olsa da literatürde vergi gelirleri ve ekonomik büyüme arasındaki ilişki net değildir. Bu çalışmada vergi gelirleri en yüksek on OECD üyesi ülke ve Türkiye’nin bulunduğu on bir ülkeli panel için vergi gelirlerinin ekonomik büyüme üzerindeki etkisi incelenmiştir. Araştırma bir panel nedensellik analizi üzerine kurulmuştur. 1971-2021 yıllarında ekonomik büyümedeki yıllık değişim ve vergi gelirlerinin GSYİH içindeki payı değişkenler olarak belirlenmiştir. Analizler Kónya (2006) panel bootstrap nedensellik testi ile gerçekleştirilmiştir. Kónya (2006) panel bootstrap nedensellik testi yatay kesit bağımlılığı dışında herhangi bir önsel test gerektirmemektedir. Bu sebeple analizler yatay kesit bağımlılığının tespit ile başlamıştır. Yatay kesit bağımlılığı analizleri CD testler ile gerçekleştirilmiştir. Test sonuçları panelde yatay kesit bağımlılığı olduğuna işaret etmiştir. Daha sonra Kónya (2006) panel bootstrap nedensellik testi uygulanmıştır. Test bulguları Belçika, Fransa ve İtalya’da ekonomik büyümeden vergi gelirlerine doğru tek yönlü nedensellik ilişkisini göstermiştir. Vergi gelirlerinden ekonomik büyümeye doğru tek yönlü nedensellik görülen ülke ise Danimarka’dır. Paneli oluşturan hiç bir ülkede çift yönlü nedensellik tespit edilmemiştir. Sonuçlar, “büyüme ve vergi” (grow and tax) hipotezinin Belçika, Fransa ve İtalya’da, “vergi ve büyüme” (tax and grow) hipotezinin ise Danimarka için geçerli olduğunu göstermiştir. Bu sonuç vergi gelirleri ile ekonomik büyüme arasındaki ilişkinin ülkeden ülkeye farklılaştığını ortaya koymaktadır. Ülkelerin vergi yapılarının, ekonomik ve konjonktürel durumlar açısından farklılık göstereceğini ve farklı vergi politikalarının ortaya çıkacağını ima etmektedir. https://doi.org/10.58830/ozgur.pub401.c1758 https://doi.org/10.58830/ozgur.pub401.c1758 165 166 Vergi Gelirleri ve Ekonomik Büyüme Üzerine Ampirik Bir Yaklaşım 1. Giriş Genel bir kabul olarak ekonomik büyüme bir ülkenin belli bir dönemde ürettiği mal ve hizmetlerin değerindeki artışları içermektedir. Ekonomik büyümeyi ölçmek için ilgili ülkenin gayri safi yurtiçi hasılasındaki (GSYİH) artış kullanılmaktadır. Arzu edilen ülkenin üretim kapasitesinde yıllık olarak ölçülen artışın istikrarlı olmasıdır. Bunun için ekonomik büyümeye neden olan ya da ekonomik büyümeyi yavaşlatan faktörlerin tespitinin doğru yapılması gerekmektedir. Herhangi bir ülkenin ekonomik gelişimi, ürettiği ve kontrol edebildiği kaynakların miktarına göre belirlenmektedir. İhtiyaç duyulan kaynakların iyi yapılandırılmış bir vergi sistemi aracılığıyla toplanacağı düşünülmektedir. Dünya genelinde ülkelerde ekonomik büyümenin düzeyi ve hızı, makroekonomik politika aracı olarak vergilendirmeyle belirlenmektedir. Gelişmiş bir vergi sistemi, hükümetin sürekli artan taleplerini karşılamak için gerekli geliri elde etmesine olanak tanımaktadır. Yani vergi, hükümetler için gerçek ve sürdürülebilir bir gelir kaynağı ve maliye politikası açısından makroekonomik yönetim için bir araçtır (Ezekwesili ve Ezejiofor, 2022:11). Vergi gelirlerini ve vergi gelirlerindeki değişikliğin ekonomik büyümeyi nasıl etkileyeceği büyüme teorileri çerçevesinde ele alınmaktadır. Vergi ile ilgili politikaların ekonomik büyümeye etkisi incelenirken öncü model neoklasik büyüme modelidir. Neoklasik modelde büyüme, yalnızca fiziksel ve beşeri sermaye birikimine bağlıdır. Uzun vadede, herhangi bir vergi yapısı bir denge sermaye/emek oranı ve işçi başına bir denge eğitim düzeyi oluşturur. Kişi başına üretimdeki herhangi bir ilave artış, basitçe dışsal teknik değişim oranından kaynaklanmaktadır. Vergi yapısının yarattığı yanlış tahsislerin büyüklüğü ne olursa olsun, vergi yapısının kişi başına düşen üretimdeki büyüme üzerinde kalıcı bir etkisi olmamalıdır. Neoklasik modelde Cobb- Douglas üretim fonksiyonunda; ölçeğe göre sabit getiri, dışsal teknoloji, sermayenin azalan getirili olması, faktörler arası ikame şansı ve bağımsız yatırım fonksiyonun bulunmadığı varsayılmaktadır. Ayrıca modelde ülkelerin uzun dönem durağan durum dengesine ulaşacağı, uzun dönem vergi politikalarını içeren hükümet politikalarının ekonomik büyümeyi etkilemeyeceği kabul edilmektedir (Saraç, 2015:23; Lee ve Gordon, 2005:1029). Tanımlandığı şekliyle neoklasik büyüme modelinde ekonomik büyüme, tasarruf ve sermaye birikimi oranları tarafından belirlenmektedir. Bu büyüme modelinde teknolojik ilerleme dışsal olarak ele alınmaktadır. 1980’lerde ortaya çıkan içsel büyüme modelleri ise teknolojik ilerlemeler aracılığıyla ekonomik büyümeyi modellemektedir. İçsel büyüme modellerinde teknolojik ilerleme, neoklasik büyüme modelinin açıklayamadığı uzun vadeli ekonomik büyümenin temel Eda Bozkurt \| 167 167 Eda Bozkurt \| 167 Eda Bozkurt belirleyicisidir. Yani teknoloji içseldir. İçsel büyüme modeli ölçeğe göre artan getirileri kabul etmektedir. Maliye politikasının rolü söz konusu olduğunda bu teori, devlet harcamalarının ve vergilendirmenin ekonominin büyümesi üzerinde hem geçici hem de kalıcı bir etkiye sahip olması gerektiğini öne sürmektedir (Maganya, 2020:208). 1. Giriş Ekonomik teoriye göre üç üretim faktörü emek, sermaye ve teknolojik ilerleme ile oluşan üretim fonksiyonunda vergiler bu faktörlere ilişkin ekonomik kararları değiştirebilir ve dolayısıyla ekonomik büyümeyi olumsuz/ olumlu yönde etkileyebilir. Bu etkiler şöyle özetlenebilir: 1) Emeğin vergilendirilmesi, ekonomik birimlerin işgücü piyasasına katılma kararını (çalışma saati ve işe alınan personel sayısı) ve daha yüksek düzeyde eğitim ve daha az girişimcilik faaliyeti arama kararını etkiler. 2) Sermayenin vergilendirilmesi hane halkının yatırım ve tasarruf kararlarını ve aynı zamanda şirketlerin yatırımların yeri ve hacmine ilişkin kararlarını da etkileyebilir. Vergi yükü yatırımın ve sermaye stokunun düşmesine neden olabilir veya yatırım düzeyini caydırarak ekonominin verimliliğini düşürebilir. 3) Emek ve sermaye verimliliğini artırmanın tek yolu olan araştırma ve geliştirmenin vergilendirilmesi, bu faaliyetlerin daha az karlı hale gelmesi nedeniyle teknolojik gelişmeleri engelleyebilir. 4) Tüketimin vergilendirilmesi, zamanlar arası kararları (tasarruf kararları, iş ve boş zaman arasındaki kararlar gibi) işgücü vergilendirmesinden daha az güçlü bir şekilde bozduğu için genellikle büyümeye daha yardımcı olabilir. 5) Vergi, sermayeyi daha verimli olan yüksek vergili sektörü terk ederek verimliliği düşük, düşük vergili sektöre zorlayacağı için sermayenin marjinal verimliliğinde düşüşe yol açabilir (Petru-Ovidiu, 2015:89; Iqbal vd. 2015:111). Vergi gelirleri ve ekonomik büyüme arasındaki ilişki teorik olarak aynı zamanda nedenselliğe dayalı olarak da ele alınmaktadır. Söz konusu nedensellik literatürde üç hipotez ile tespit edilmektedir. İlk hipotezde, vergi gelirlerinden ekonomik büyümeye doğru tek yönlü bir nedensellik ilişkisi “vergi ve büyüme” (tax and grow) hipotezi olarak adlandırılmaktadır. Teorinin savunucusu Friedman (1978) vergi oranlarının veya vergi gelirini artırmanın, büyümeyi teşvik edeceği daha fazla mali alana yol açacağını savunmaktadır. İkinci hipotez, “büyüme ve vergi” (grow and tax) hipotezi ekonomik büyümeden vergi gelirlerine doğru tek yönlü bir nedenselliği ima etmektedir. Peacock ve Wiseman (1979), Keynesyen büyüme teorisini uygulayarak hükümet harcamalarının neden olduğu büyümenin krizler Vergi Gelirleri ve Ekonomik Büyüme Üzerine Ampirik Bir Yaklaşım 168 nedeniyle artabileceğini ve artan seviyelerin harcama artışını kriz bittikten sonra bile devam ettirdiğini öne sürmektedir. Bu ilişkide başlangıçta hükümet harcamalarını zorlayan ciddi krizlerin, vergi gelirlerindeki büyüme oranından daha fazla ekonomik büyüme oranına neden olduğu varsayılmaktadır. Esas fikir krize bağlı olarak ortaya çıkan orijinal vergi gelirlerindeki artışların vergi politikalarında kalıcı bir özellik haline gelmesidir. Son hipotez ise ekonomik büyüme ile vergi geliri arasında çift yönlü bir nedensellik olduğunu öne sürmektedir. Barro’nun (1979) vergi düzleştirme modeline (tax smoothing model) dayanan mali senkronizasyon hipotezidir. Bu hipotez, hükümetin vergi harcamalarının büyümeyi tetiklediği ve vergi gelirini maksimize etme kararlarının eş zamanlı olarak alındığını açıklamaktadır (Dzingirai ve Tambudzai, 2014:14). 1. Giriş Yukarıda sunulan teorik önemi ve herhangi bir vergi değişikliğinin ekonomik faaliyetleri etkileyebileceği fikrinden hareketle çalışmada vergi gelirleri en yüksek 10 Ekonomik Kalkınma ve İş Birliği Örgütü (OECD) üyesi ülke ve Türkiye’nin yer aldığı panel için 1971-2021 yılları arasında nedensellik araştırması yapılmıştır. Araştırma genel teorik bilgilerin verildiği giriş başlığının ardından literatür özeti ile devam etmiştir. Daha sonra veri seti ve yöntem tanıtılmış, ardından ekonometrik analiz sonuçları sunulmuştur. Çalışma sonuç başlığı ile tamamlanmıştır. 2. Vergi Gelirlerinin Ekonomik Büyümeye Etkisi Literatür Özeti Ulusal ve uluslararası yazında vergi gelirlerinin iktisadi büyümeye etkisini araştıran pek çok çalışma olduğu bilinmektedir. Tablo 1 yardımıyla bu çalışmalara ait özet bir tartışma sunulmuştur. Çalışmaların özellikleri şu şekilde sıralanabilir: Araştırmalarda genel bir eğilim olarak ampirik yöntemlere sıkça başvurulmuştur. Bu yöntemlerden bazıları zaman serileri bazıları ise panel verileri içermektedir. Türkiye’de yapılan çalışmaların görece olarak daha fazla zaman serisi teknikleriyle tasarlandığı söylenebilir. Panel veri ile yapılan araştırmalar daha az sayıda olup çalışmanın bu anlamda alana katkı sağlayacağı beklenmektedir. Ekonometrik teknikler kullanılarak yapılan çalışmalarda ülke, bölge, dönem, değişken ve yöntemlerin çeşitlilik gösterdiği görülmektedir. Bulguları bakımından bazı çalışmalarda vergilerin ekonomik büyüme performansıyla pozitif, bazılarında ise negatif ilişki içinde olduğu tespit edilmiştir. Öte yandan bazı araştırmalarda ise ekonomik büyüme ile vergi gelirleri arasındaki ilişki için kanıt elde edilmemiştir. Yine nedensellik sonucu sunan çalışmalar tek yönlü, çift yönlü ilişkiye ya da nedensel ilişkisizliğe işaret ederek çeşitlilik göstermektedir. Özetle vergi gelirleri ve iktisadi büyüme arasındaki ilişki için yapılan çalışmalarda çelişkili sonuçlar bulunmaktadır. Eda Bozkurt \| 169 Eda Bozkurt \| 169 Eda Bozkurt \| 169 Tablo 1. Ekonomik Büyüme ve Vergi Gelirleri Literatür Taraması Yazar/Yılı Ülke/Dönem Yöntem Sonuç Siverekli Demircan (2003) - Deskriptif Dolaysız vergiler; ekonomik büyümeyi olumlu etkilerken, dolaylı vergiler, tasarrufların teşviki, tüketimin azaltılması yurtiçi sanayinin gelişmesine katkı sağlaması yönünden önemlidir. Ünlükaplan ve Arısoy (2011) Türkiye/ 1968- 2006 Johansen ve Juselius (1990) Eşbütünleşme Testi, Vektör Otoregresyon (VAR), Granger Nedensellik Testi Vergi yükü ile ekonomik büyüme eşbütünleşik olup, iktisadi büyüme vergi yükü arasında çift yönlü nedensellik vardır. Stoilova ve Patonov (2013) 27 Avrupa Birliği üyesi ülke/ 1995- 2010 En Küçük Kareler Yöntemi (OLS) Dolaysız vergiler ekonomik büyümeyi desteklemede daha etkindir. Yaraşır Tülümce (2013) Türkiye/ 1988- 2010 Johansen Eşbütünleşme ve Granger Nedensellik Testleri Dolaylı vergiler ve büyüme arasında pozitif ilişki vardır. Nedensellik ilişkisi yoktur. Dzingirai ve Zachary (2014) Zimbabve/ 1980- 2012 Granger Nedensellik, Johansen Eşbütünleşme Testi ve Vektör Hata Düzeltme Modeli Vergi gelirleri ekonomik büyüme üzerinde etkili değildir. Iqbal vd. (2015) Pakistan/ 1979- 2010 Çoklu Doğrusal Regresyon Modeli Vergilendirmenin ekonomik büyüme üzerinde olumlu etkisi vardır. Petru-Ovidiu (2015) 6 ülke/ 1995-2012 Panel Regresyon Modeli Dolaysız vergiler ekonomik büyümeyle negatif, dolaylı vergiler ise pozitif ilişkilidir. Saraç (2015) Türkiye/ 1969- 2013 Markov Rejim Değişim Dolaysız vergiler büyümeyi negatif, dolaylı vergiler ise pozitif etkilemektedir. Terzi ve Yurtkuran (2016) Türkiye/ 1980- 2013 VAR Modeli ve Toda- Yamamoto (TY) Nedensellik Testi Dolaysız vergiler ile GSYİH arasında çift yönlü nedensellik vardır. Dolaysız vergi oranlarındaki azalma GSYİH’yi pozitif yönde etkilemektedir. Tablo 1. 2. Vergi Gelirlerinin Ekonomik Büyümeye Etkisi Literatür Özeti Ekonomik Büyüme ve Vergi Gelirleri Literatür Taraması 170 \| Vergi Gelirleri ve Ekonomik Büyüme Üzerine Ampirik Bir Yaklaşım Demir ve Sever (2017) 11 OECD ülkesi/1980-2014 Westerlund Panel Eşbütünleşme Testi Uzun dönemde dolaysız vergiler geliri azaltırken, kısa dönemde dolaysız vergiler-dolaylı vergiler ve toplam vergiler geliri negatif etkilemektedir. Kolçak ve Arpa (2017) Türkiye/ 1975- 2016 Gecikmesi dağıtılmış otoregresif (ARDL) sınır testi Dolaylı vergiler ile büyüme aynı yönde hareket ederken dolaysız vergiler ve servet vergileri zıt yönde hareket etmektedir. Sandalcı ve Sandalcı (2017) OECD ülkeleri/ 1990-2014 Modifiye Edilmiş En Küçük Kareler Yöntemi (FMOLS) ve Dumitrescu- Hurlin (2012) Nedensellik Test Ekonomik büyüme ve dolaylı vergiler arasında pozitif, toplam vergiler ve dolaysız vergiler arasında ise negatif ilişki vardır. Su ve Nguyen (2017) 82 gelişmiş ve gelişmekte olan ülke/2000-2015 Sistem Genelleştirilmiş Momentler Metodu (GMM) Vergi gelirlerinin ekonomik etkisi ülke gruplarına göre değişmektedir. Ali vd. (2018) Kenya/ 1980-2017 OLS Vergi gelirlerinin ekonomik büyüme üzerinde anlamlı pozitif etkisi vardır. Egbunike vd. (2018) Nijerya ve Gana/ 2000-2016 OLS ve Granger Nedensellik Testleri Vergi gelirlerinin GSYİH üzerinde anlamlı pozitif etkisi vardır. Oboh vd. (2018) Nijerya, Gana, Sierra Leone, Benin ve Burkina Faso/2000-2015 Görünüşte İlgisiz Regresyon Modeli Dolaylı vergiler ekonomik büyümeyi artırır. Owino (2018) Kenya/ 1973-2010 Johansen Eşbütünleşme Testi ve Hata Düzeltme Modeli Dolaysız vergi ile ekonomik büyüme arasında negatif, dolaylı vergiler ile ekonomik büyüme arasında pozitif ilişki vardır. Akıncı (2019) Türkiye/2006:Q1- 2018:Q3 Maki (2012) Çoklu Yapısal Kırılmalı Eşbütünleşme Testi Ekonomik büyüme ile vergi gelirleri eşbütünleşiktir. Boğa (2020) Türkiye/ 1965- 2018 Doğrusal Olmayan ARDL (NARDL) Eşbütünleşme Testi Vergi gelirlerinin ekonomik büyüme üzerindeki pozitif etkisi vergi indirimlerinin oluşturduğu pozitif etkiden daha güçlüdür. Eda Bozkurt \| 171 Maganya (2020) Tanzanya/ 1996- 2019 ARDL Vergi gelirlerinin türüne göre ekonomik büyüme etkisi değişkenlik göstermektedir. Ülkü ve Açıkgöz (2020) Türkiye/ 1980- 2017 ARDL Vergi ile ilgili göstergelerden büyümeye kuvvetli nedensel ilişki yoktur. Bıçaksız ve Gürler Hazman (2021) - Deskriptif Dolaylı vergiler ekonomik büyümeyi olumlu etkilemektedir. Korkmaz vd. (2022) 9 OECD ülkesi/ 2010-2019 Panel Nedensellik Testi Vergi gelirleri ekonomik büyümeye ile tek yönlü nedensellik ilişkisi içindedir. Özen vd. (2022) Türkiye/ 2006:1- 2020:1 Johansen Eşbütünleşme, Kanonik Eşbütünleşik Regresyon (CCR) Testi, Dinamik En Küçük Kareler Yöntemi (DOLS), FMOLS Dolaylı ve dolaysız vergi gelirleri ekonomik büyümeyi pozitif etkilemektedir. Ho vd. (2023) 29 gelişmekte olan ülke/2000-2020 Sabit Etkiler Modeli Vergi gelirleri ekonomik büyümeyi olumlu etkilemektedir. 3. Veri Seti ve Ekonometrik Yöntem Vergi gelirlerinin ekonomik büyüme etkisinin incelendiği bu çalışmada vergi gelirleri 2021 yılı itibariyle en yüksek 10 OECD ülkesi ele alınmıştır. Bu ülkeler Avusturya, Belçika, Danimarka, Finlandiya, Fransa, Almanya, İtalya, Hollanda, Norveç ve İsveç’ten oluşmaktadır. Ayrıca panele 11. ülke olarak Türkiye eklenmiştir. 1971-2021 yılları için ekonomik büyüme yıllık ekonomik büyüme oranı ile temsil edilmiş ve Dünya Bankası veri tabanından elde edilmiştir. Vergi gelirleri ise vergi gelirlerinin GSYİH’ye oranı olarak OECD verilerinden oluşturulmuştur. Çalışmada Kónya (2006) tarafından geliştirilen panel bootstrap nedensellik testi uygulanmıştır. Bootstrap panel nedensellik testini için birim kök testi ya da eşbütünleşme testi gibi zaman serisi özelliklerinin incelenmesi gerekmemektedir. Kónya (2006) panel bootstrap nedensellik testi için gerekli tek ön koşul olan paneli oluşturan yatay kesit birimler arasındaki 172 Vergi Gelirleri ve Ekonomik Büyüme Üzerine Ampirik Bir Yaklaşım bağımlılığın tespitidir (Altıner vd., 2018:29). Yatay kesit bağımlılığı için cd Lm1 (Breusch, Pagan 1980), cd LM2 (Pesaran 2004 CDlm), cd LM (Pesaran 2004 CD), Bias-adjusted CD test raporlanmıştır. Daha sonra Konya (2006) panel bootstrap nedensellik testi yapılmıştır. Kónya (2006) panel bootstrap nedensellik testi, görünürde ilişkisiz regresyonlara ve yatay kesit birime özgü spesifik olarak elde edilen bootstrap kritik değerlere sahip Wald istatistiği ile tahmin yapmaktadır. Tahminler aşağıdaki denklem (1) ve (2) ile gerçekleştirilmektedir. y ve x değişkenleri, t dönemi (t=1,…,T), i ülkeleri (i=1,…,N), l gecikme uzunluğunu , beyaz gürültülü hata terimlerini göstermektedir. Denklemler farklı önceden belirlenmiş değişkenlere sahip olup, bireysel regresyonlar arasındaki tek olası ilişki sistemdeki eşanlı korelasyon olduğundan, bu denklem setleri görünürde ilişkisiz regresyonları ifade etmektedir. Ayrıca ülkeye özgü bootstrap kritik değerler ele alındığından birim kök testi gibi zaman serisi özelliklerine bakılmadan ve değişkenlerinin durağan olması gerektiği varsayımı söz konusu değildir. Bu yaklaşımda paneli oluşturan yatay kesit birimlerin tümü için ortak hipotezler gerekmez. Yatay kesit birimler arasında eşanlı korelasyon, panel veri tarafından sağlanan ilave bilgilerin kullanılmasına imkan tanır. Ek olarak uygun gecikme yapısı dışında herhangi bir ön teste ihtiyaç yoktur. Nedensellik testi kritik olarak gecikme yapısına bağlı olup, tahmin yapılmadan gecikme sayısı belirlenmelidir. Maksimum gecikme sayısına bu testte Schwartz ve Akaike Bilgi Kriterini minimum yapacak optimal gecikme uzunluğunun 1-4 arasında olduğu varsayılmaktadır (Kónya, 2006: 979-982). ve Eda Bozkurt \| 173 Sonuç olarak bu teste göre nedensellik ilişkisinin belirlenmesi amacıyla Wald istatistikleri bootstrap yöntemiyle elde edilen her yatay kesit birim için elde edilen kritik değerlerle karşılaştırılır. 3. Veri Seti ve Ekonometrik Yöntem Herhangi bir yatay kesit birim için Wald istatistiğinin bootstrap kritik değerden büyük olduğu tespit edildiğinde değişkenler arasında nedensellik ilişkisi olduğu ya da tersi durumda herhangi bir nedensellik ilişkisinin olmadığı söylenmektedir (Altıner vd.2018:30). 4. Ampirik Sonuçlar İlk aşamada cd Lm1 (Breusch,Pagan 1980), cd LM2 (Pesaran 2004 CDlm), cd LM (Pesaran 2004 CD), Bias-adjusted CD test ile yapılan yatay kesit bağımlılığı test sonuçları Tablo 2’de sunulmuştur. Tablo 2. Yatay Kesit Bağımlılığı Test Sonuçları Test Birinci Model İkinci Model cd Lm1 (Breusch,Pagan 1980) 994.489 (0.000) 684.479 (0.000) cd LM2 (Pesaran 2004 CDlm) 89.577 (0.000) 250.711 (0.000) cd LM (Pesaran 2004 CD) 29.542 (0.000) 51.791 (0.000) Bias-adjusted CD test 4.107 (0.000) 1.397 (0.000) Tablo 2. Yatay Kesit Bağımlılığı Test Sonuçları Hem birinci modelde hem de ikinci modelde sıfır hipotezi tüm testlerde %1 anlamlılık düzeyinde reddedilmiştir. Bu sonuç her iki modelde yatay kesit Vergi Gelirleri ve Ekonomik Büyüme Üzerine Ampirik Bir Yaklaşım 174 bağımlılığı olduğunu göstermiştir. Yatay kesit bağımlılığının varlığı Kónya (2006) panel bootstrap nedensellik testi ile vergi gelirleri ve ekonomik büyüme arasındaki ilişkinin araştırılması için imkân sunmuştur. Bu amaçla yapılan Kónya (2006) panel bootstrap nedensellik testi sonuçları Tablo 3’te verilmiştir. Tablo 3. Kónya (2006) Bootstrap Nedensellik Testi Sonuçları Ülkeler Birinci Model İkinci Model Wald İstatistiği B.K.D. %1 B.K.D. %5 B.K.D. %10 Wald İstatistiği B.K.D. %1 B.K.D. %5 B.K.D. %10 Avusturya 4.253 8.572 7.172 4.807 0.074 15.682 12.220 9.892 Belçika 9.768 9.547 7.217 5.052 0.185 20.608 16.271 13.783 Danimarka 0.439 7.448 4.303 3.685 4.270 4.168 2.729 1.896 Finlandiya 2.652 13.645 10.923 7.887 0.686 12.055 8.725 7.273 Fransa 14.971 8.380 6.530 4.568 5.450 19.981 13.880 12.022 Almanya 0.255 10.165 5.483 3.888 1.540 4.927 3.827 2.110 İtalya 47.914 18.317 9.151 7.808 0.063 16.231 11.504 9.793 Hollanda 1.318 9.486 4.070 2.710 0.005 2.976 2.108 1.492 Norveç 2.997 11.146 7.135 3.644 1.160 4.489 2.618 2.278 İsveç 0.073 11.918 6.068 4.803 2.476 10.605 4.113 3.388 Türkiye 0.305 10.396 5.266 3.467 0.007 6.704 5.694 4.202 Not: B.K.D. Bootstrap Kritik Değerleri ifade etmektedir. Bootstrap kritik değerler için 10.000 döngüyle Monte Carlo Simülasyonu kullanılmıştır. Not: B.K.D. Bootstrap Kritik Değerleri ifade etmektedir. Bootstrap kritik değerler için 10.000 döngüyle Monte Carlo Simülasyonu kullanılmıştır. Not: B.K.D. Bootstrap Kritik Değerleri ifade etmektedir. Bootstrap kritik değerler için 10.000 döngüyle Monte Carlo Simülasyonu kullanılmıştır. Tablo 3’teki sonuçlara göre paneli oluşturan 11 ülkeden Belçika, Fransa ve İtalya’da %1 önem düzeyinde ekonomik büyümeden vergi gelirlerine doğru doğru nedensellik ilişkisi bulunmaktadır. Bu üç ülkede dışında diğer ülkelerde ekonomik büyümeden vergi gelirlerine doğru nedensellik tespit edilememiştir. Vergi gelirlerinden ekonomik büyümeye doğru tek yönlü nedensellik görülen tek ülke %1 önem düzeyinde olmak üzere Danimarka’dır. Danimarka dışında diğer ülkeler için vergi gelirlerinden ekonomik büyümeye doğru nedensel ilişki bulunamamıştır. 4. Ampirik Sonuçlar Öte yandan paneli oluşturan hiç bir ülkede çift yönlü nedensel bağlantı görülmemiştir. Sonuçlar, “büyüme ve vergi” (grow and tax) hipotezinin Belçika, Fransa ve İtalya’da, “vergi ve büyüme” (tax and grow) hipotezinin ise Danimarka için geçerli olduğunu göstermiştir. Eda Bozkurt \| 175 Eda Bozkurt \| 175 SONUÇ Ekonomistlerin en önemli uğraşlarından biri ülkelerdeki farklı büyüme oranlarının arkasındaki nedenleri tespit etmektir. Vergi gelirlerinin de ülkelerin üretim kapasiteleri ve ekonomik büyüme oranlarındaki farklılığı etkileyen önemli bir büyüklük olduğu bilinmektedir. Vergi politikaları ülke içinde daha yüksek tasarruflar, fiyat istikrarı, kaynak tahsisi, üretim ve tüketim düzeyinin kontrol altına alınması gibi etkilerle ekonomik büyümeyi değiştirmektedir. İşte burada ülkelerdeki vergi gelirlerinin ekonomik büyümeyi nasıl değiştirdiği inceleme konusu olmaktadır. Bu çalışmada 11 ülkede 1971-2021 döneminde Kónya (2006) panel bootstrap nedensellik testi aracılığıyla vergi gelirlerinin ekonomik büyüme ile ilişkisi incelenmiştir. Sonuçlar, Belçika, Fransa ve İtalya’da büyüme ile vergi gelirlerinin tek yönlü nedensel ilişki içinde olduğunu ortaya koymuştur. Yine bulgular vergi gelirlerinin ekonomik büyümeyi ise tek yönlü olarak Danimarka’da etkilediğini göstermiştir. Ülke ekonomilerine vergilerin olumlu yansıması için büyümeyi odak noktasına alan bir makroekonomik politikanın oluşturulması önemlidir. Bunun için vergiler ve vergi artışları ekonomideki işleyişi bozmayacak şekilde tasarlanmalıdır. Vergiler istihdamı destekleyici, yenilikçilik ve girişimciliği geliştirecek kültürde olmalıdır. Vergiler ile ilgili şeffaf uygulamalar iyi denetim ve buna uygun mekanizmaların inşasını gerektirmektedir. Bu çerçevede vergi kaçakçılığı ve vergiden kaçınma konuları üzerine gidilmelidir. Vergiler ülkelerin iç kaynaklarını harekete geçirmede, gelir dağılımı adaletini sağlamada etkin bir araç olarak düşünülmelidir. Son olarak bu çalışmanın önemli sınırlılığı vergi gelirlerinin yapısını dışlamış olmasıdır. 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Vergi yükü ve yapısı ile iktisadi büyüme arasındaki dinamik etkileşimler üzerine uygulamalı bir analiz. ODTÜ Ge- lişme Dergisi, 38(Nisan), 71-100. World Bank (2023). World Development Indicators. https://databank.wor- ldbank.org/source/world-development-indicators, Date Accessed: 10.11.2023. Yaraşır Tülümce, S. (2013). Türkiye’de vergi gelirleri ve büyüme arasındaki ilişkinin ampirik analizi (1988-2010). Gümrük ve Ticaret Dergisi, 1(2), 56-64. Özet Emek arzı ve emek talebinin buluştuğu ücret düzeyinin, çalışma koşullarının şekillendiği bir organizasyon bütünü olarak ifade edilen işgücü piyasası, küreselleşmenin yarattığı sonuçlardan etkilenmiştir. Özellikle 1980’den sonra yaşanan neoliberalizm olarak adlandırılan ekonomik ve siyasi paradigma değişikliği, artan işsizlik sorunu ve rekabet koşullarının değişmesi işgücü piyasası için de farklı piyasa taleplerine cevap verebilecek dönüşümleri kaçınılmaz kılmıştır. Klasik istihdam ilişkisinin yerini giderek artan bir biçimde a-tipik istihdam olarak da nitelendirilen esnek çalışma biçimleri almıştır. İstihdamda esnekliğin getirilmesi ile işgücü piyasasında yeni fırsat ve tehditler de ortaya çıkmıştır. Esneklik çalışma yaşamında işlerin niteliğini, istihdam türlerini ve çalışma sürelerinde farklılaşmayı getirmiştir. Esneklik; üretimde esneklik, yönetimde esneklik ve esnek istihdam (esnek çalışma) olarak işgücü piyasasında farklı şekillerde detaylanmaktadır. Bu çalışmada konu, esnek istihdam çerçevesinde sınırlandırılmıştır. Bu kapsamda esnek istihdam uygulamalarından bir olan “geçişi iş ilişkisi” Türk İş Hukuku mevzuat hükümleri çerçevesinde değerlendirilmiştir. Geçici iş ilişkisi, esasen 4857 sayılı İş Kanunu’nun 7. maddesinde düzenlemiştir. İlgili maddeye göre işveren, işçisinin yazılı onayını almak koşulu ile başka bir işverene geçici olarak devredebilmektedir. Ancak yapılan bir değişiklikle geçici iş ilişkisinin kapsamı genişletilmiş olup, geçici iş ilişkisi iki şekilde uygulama alanı bulmuştur. Birincisi “meslek edinilmemiş geçici iş ilişkisi”dir. İkinci ise, “meslek edinilmiş geçici iş ilişkisi” olarak ifade edilmiştir. Bu kapsamda geçici iş ilişkisi, Özel İstihdam Büroları aracılığı ile yapılmaktadır. Çalışmada geçici iş ilişkisinin her iki uygulama alanına yer verilmiş olup meslek edinilmiş geçici iş ilişkisi detaylandırılarak işgücü piyasası açısından etkileri değerlendirilmiştir. https://doi.org/10.58830/ozgur.pub401.c1759 https://doi.org/10.58830/ozgur.pub401.c1759 179 180 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi 1. Esnek İstihdam ve Esnek İstihdam Türleri Üretim sistemleri ve buna bağlı hızla değişen istihdam şekilleri ve endüstri ilişkileri, sistemde meydana gelen yeni dönüşümleri beraberinde getirmiştir. Post-fordist üretim sistemi ile başlayan esneklik, piyasa belirsizliği ve piyasasının katı sistemine bir çözüm anlayışı olarak görülmüş ve günümüze kadar değişen varyasyonlarda daha da görünür olmuştur. Beş farklı işgücü piyasası esnekliğinden bahsetmek mümkündür. Bunlar; ücret esnekliği, sayısal esneklik, fonksiyonel/işlevsel esneklik, coğrafi esneklik ve esnek çalışma modelleri olarak farklı boyutlarda şekillenmiştir. Ayrıca, işgücü piyasasında esneklik; esnek üretim, esnek yönetim ve esnek çalışma olarak üç grupta değerlendirilmektedir. Çalışma dahilinde konu esnek çalışma/esnek istihdam ile sınırlı tutulmuştur. 1.1. Esneklik ve Esnek İstihdam Kavramları 1.1.1. Kavram Olarak Esneklik Parlak ve Özdemir (2011: 6), Esneklik Kavramı ve Emek Piyasalarında Esneklik, In Journal of Social Policy Conferences. Kaynak: Meulders ve Wilkin’den Akt. Parlak ve Özdemir (2011: 6), Esneklik Kavram ve Emek Piyasalarında Esneklik, In Journal of Social Policy Conferences. Kaynak: Meulders ve Wilkin’den Akt. Parlak ve Özdemir (2011: 6), Esneklik Kavramı ve Emek Piyasalarında Esneklik, In Journal of Social Policy Conferences. 1.1.1. Kavram Olarak Esneklik Son yıllarda ekonomi ve sosyal politika literatüründe sıklıkla kullanılan ve tartışmaya konu olan kavramlardan bir esneklik olmuştur. Fordist dönemin istikrarlı, tam zamanlı, tam istihdam ve tam refah hakları ile ortaya çıkan krizinden bugüne “esneklik”, hızla yaygınlaşmıştır (Barbieri, 2009:621). Es- neklik farklı boyutları ve türleri içermesi nedeniyle tek ve yalın bir tanımlama yapmak mümkün değildir. Esneklik, “kısa bir süre içinde çok az bir maali- yet ve çabayla yahut da performans ile değişebilme veya uyum sağlayabilme yeteneği olarak tanımlanmaktadır” (Parlak ve Özdemir, 2011:3). Genel bir ifade ile “esneklik sık kullanılan her duruma uygulanabilen bir kavramdır. Esneklik, bir sitem veya alt sistemin sisteme yönelik çeşitli etkenlerle uyum- lu olarak tepki gösterebilmesini ifade etmektedir”. Bu açıdan esneklik kav- ramının temel unsuru değişebilirlik olmaktadır (Tokol, 2008:122). Ayrıca esneklik kimi zaman bir ihtiyaç, uygulaması zorunlu bir yöntem ve maliyet azaltma unsuru iken, kimi zaman da kuralsızlaşma, eğretilik, güvencesizlik olarak ifade edilmiştir. Kuşkusuz kavrama yüklenen yorum ne olursa olsun işgücü piyasasında esneklik uygulamaları hızla yaygınlaşmaktadır. Bu neden- le esnek uygulamaların ortaya çıkardığı sorunlara karşı önlem geliştirmek, düzenlemeler yapmak da ayrıca önem kazanmıştır. Ekonomik, sosyal ve teknolojik değişimler firmaların işlerini esnek bir şekilde sürdürebilmelerini zorunlu kılmaktadır. Özellikle teknik ilerleme sadece yeni ürünlere ve üretim yöntemlerine ihtiyacı artırmakla kalmıyor, aynı zamanda nicelik ve niteliğe ilişkin istihdam ayarlamalarını da zorunlu kılmaktadır (Pfeifer, 2005:405). İşgücü piyasasında esneklik, “çağın getirdiği yeniliklere, ilerleyen teknolojilere beklenen uyumun sağlanması” 181 Özlem Demir anlamına gelir. Kısaca esneklik, işletmenin değişen şartlara en kısa sürede uyum yeteneğini sağlanıp, rekabet gücünün artırılmasını hedeflemektedir (Öztürkoğlu, 2013: 110). Esasen esneklik, 1970 krizi sonrası gelişen yeni teknolojiler, bilişim sisteminde hızlı ilerleme ile neoliberal politikaların uygulamaya girmesi ile standart ve kurallardan her türlü sapma durumunu ifade etmektedir. Literatürde çok sayıda esneklik türleri söz konusudur. Temel ayrım; içsel ve dışsal esneklik ile sayısal ve işlevsel esnekliktir. Bu ayrıma göre esneklik türleri Tablo-1’de gösterilmiştir. Tablo 1. İşgücü Piyasasında Esneklik Türleri Sayısal Esneklik İşlevsel Esneklik İçsel Esneklik Yarı zamanlı çalışma, esnek zaman düzenlemesi, yıllara göre ayarlanmış çalışma saatleri, çalışma süreleri muhasebesi Çoklu göre, iş zenginleştirme, çoklu vasıflı, takım çalışması. Dışsal Esneklik Süresi belirli iş sözleşmesi, serbest çalışan, kiralık işçi bulma kurumları, düzensiz iş, mevsimlik çalışma, geçici işten çıkarma. Taşeron (alt işveren) işçileri, dışardan tedarik ve serbest lisans. Kaynak: Meulders ve Wilkin’den Akt. Parlak ve Özdemir (2011: 6), Esneklik Kavramı ve Emek Piyasalarında Esneklik, In Journal of Social Policy Conferences. Tablo 1. İşgücü Piyasasında Esneklik Türleri Kaynak: Meulders ve Wilkin’den Akt. 1.1.2. Kavram Olarak Esnek İstihdam Esnek istihdam, işverene çalışanların çalışma saatlerini kısa sürede değiştirme ve/veya nispeten düşük maliyetlerle işten çıkarma olanağı tanıyan standart dışı istihdam türlerini ifade etmektedir (Olsthoorn, 2015:25). Genel manada esnek istihdam, işverenlerin işgücünü ihtiyaçlara ve değişen koşullara göre düzenleyebilmesi ile ilgilidir. Esnek istihdam güvencesiz, eğreti istihdam olarak da adlandırılır. Güvencesiz istihdam edilenler düşük ücretli, yüksek işsizlik riski olan, işsizlik yardım hakları düşük ve ek gelir imkanları sınırlı esnek sözleşmelerle çalışanlar olarak tanımlanır (Olsthoorn, 2015:174). Benzer şekilde eğreti istihdam da “standart istihdam ilişkisinden uzak, güvencesiz, kötü çalışma koşullarının olduğu, işten çıkarma ve çıkarılma riskinin yüksel olduğu, ekonomik ve sosyal kırılganlığın hat safhada olduğu bir yapı” olarak tanımlanmaktadır (Temiz, 2004). Ücretleri düzenli olarak azalmak, iş yükünü artırmak, maliyeti düşürmek için esnek istihdam bir araç konumunda olduğu belirtilmektedir. Özellikle Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi 182 işçi sınıfının baskılanması ve sendikal örgütlenmenin engellenmesi amacıyla da kullanılmaktadır (Belek, 1994:27). Bu nedenle esnek istihdam çalışanların güvencesizlik ve istikrarsızlık yaşamasına neden olabileceğinden, esnek istihdam uygulanırken çalışanın hak ve ihtiyaçlarının gözetilmesi önemlidir. Esnek istihdamın yaygınlaşması ile son yıllarda “esneklik” ve “güvence” kavramlarının birleştirilmesi ile yeni bir kavram olan “güvenceli esneklik” kullanılmaya başlanmıştır. Avrupa İstihdam Stratejisi’nde işgücü piyasasında esneklik ve güvencenin aynı anda gerçekleştirilmesi bir strateji olarak kabul edilmiştir (European Commission, 2023). Türkiye’de de Avrupa İstihdam Stratejisine benzer şekilde “Ulusal İstihdam Stratejisi” hazırlanmış, işgücü piyasasında güvence ve esnekliğin sağlanması konusunda adımlar atılmıştır. Ancak ekonomik istikrarsızlık, küçük ölçekli işletmelerin ağırlıklı yapısı, kayıt dışı istihdamın yaygın olması, sosyal diyaloğun zayıflığı gibi unsurlar güvenceli esnekliğin önünde engeller oluşturmaktadır (Kuzgun, 2012:47). 1.2. Esnek İstihdam Türleri İşletmelerin değişen koşullara ve ihtiyaca göre işgücü ayarlarken kullandığı çeşitli araçlar, esnek istihdam türleri ile şekillenmektedir. Uygulama açısından esnek çalışma türlerini hem süre hem de istihdam biçimi bakımından temel iki ayrımla detaylandırmak mümkündür. Çalışma süresine göre esnek çalışma türleri; kısmi süreli çalışma (part-time çalışma, yarı zamanlı çalışma), çağrı üzerine çalışma, belirli süreli çalışma, vardiyalı çalışma, kısa çalışma, kayan iş süresi, iş paylaşımı, denkleştirme uygulaması, telafi çalışmasıdır. İstihdam biçimleri bakımından esnek çalışma türleri ise; evde çalışma, tele (e-çalışma) çalışma, geçici iş ilişkisi, alt işverenlik uygulamasıdır. 1.2.1. Çalışma Süresine Göre Esnek İstihdam Türleri 1.2.1. Çalışma Süresine Göre Esnek İstihdam Türleri Çalışma süresine göre esnek istihdam türlerinden en yaygın şekliyle uygulamada olan kısmi süreli çalışmadır. En temel tanımı ile kısmi süreli çalışma (yarı zamanlı çalışma, part-time çalışma), “karşılaştırılabilir tam zamanlı çalışmaya göre daha kısa olan çalışma saatleri”ni ifade etmektedir (ILO, 2014). Başka bir ifade ile kısmi süreli çalışma, çalışma saatlerinin normalden az olduğu düzenli ücretli istihdam olarak tanımlanmaktadır (Kalleberg, 2000:343). Yavuz’a (1995:38) göre kısmi süreli çalışmanın üç unsuru vardır: Birincisi, kısmi süreli çalışma normal çalışma süresinden daha az olmalı, ikicisi yapılan bu çalışmanın düzenli ve sürekli olması, üçüncüsü ise, bu çalışmanın gönüllü yapılmasıdır. 4857 sayılı İş Kanun’un 13.maddesi “işçinin normal haftalık çalışma süresinin, tam süreli iş sözleşmesiyle çalışan emsal işçiye göre önemli ölçüde daha az belirlenmesi durumunda sözleşme kısmi süreli iş sözleşmesidir” hükmü ile kısmi çalışmanın dayanağı 183 Özlem Demir Özlem Demir oluşturulmuştur. Esnek çalışmanın farklı bir türü olan “çağrı üzerine çalışma” literatürde sıfır saat sözleşme veya stand-by çalışma olarak adlandırılmakta ve oldukça düşük iş güvencesi sağlamaktadır. Bu nedenle bazı ülkelerde iş güvencesi sağlamak ve çalışan haklarını korumak için yasal düzenleme yapılmıştır. Türkiye’de 4857 sayılı İş Kanunu’nda düzenlenmiştir. Buna göre; “Yazılı sözleşme ile işçinin yapmayı üstlendiği işle ilgili olarak kendisine ihtiyaç duyulması halinde iş görme ediminin yerine getirileceğinin kararlaştırıldığı iş ilişkisi, çağrı üzerine çalışmaya dayalı kısmi süreli bir iş sözleşmesidir. Hafta, ay veya yıl gibi bir zaman dilimi içinde işçinin ne kadar süreyle çalışacağını taraflar belirlemedikleri takdirde, haftalık çalışma süresi yirmi saat kararlaştırılmış sayılır. Çağrı üzerine çalıştırılmak için belirlenen sürede işçi çalıştırılsın veya çalıştırılmasın ücrete hak kazanır. İşçiden iş görme borcunu yerine getirmesini çağrı yoluyla talep hakkına sahip olan işveren, bu çağrıyı, aksi kararlaştırılmadıkça, işçinin çalışacağı zamandan en az dört gün önce yapmak zorundadır. Süreye uygun çağrı üzerine işçi iş görme edimini yerine getirmekle yükümlüdür. Sözleşmede günlük çalışma süresi kararlaştırılmamış ise, işveren her çağrıda işçiyi günde en az dört saat üst üste çalıştırmak zorundadır” (İş K. 14.md.). Süreye bağlı şekillenen diğer bir esnek çalışma modeli “kayan iş süresi”dir. Genel bir ifade ile işçinin günlük ve belirli çalışma süresine bağlı kalmaksızın işe başlama, ara dinlenme, işi bitirme gibi günlük çalışma süresinin uzunluğunu değişik biçimde belirleme olanağına sahip olduğu bir çalışma modelidir (Soyer, 1994:183-184). Kayan iş süresi, basit kayan iş süresi ve nitelikli kayan iş süresi olarak iki şekilde uygulanmaktadır. Basit kayan iş süresi uygulamasında, çekirdek zaman dilimi içinde işçi işe başlama ve işi bitirme saatini kendisi belirlemektedir. 1.2.1. Çalışma Süresine Göre Esnek İstihdam Türleri Nitelikli kayan iş süresinde, işçinim sadece günlük zaman diliminde değil hafta ya da ay bazında eksik veya fazla çalışma süresini de denkleştirme olanağına sahiptir (Tuncay, 1995:64). Çalışma yaşamındaki değişime paralel şekilde ortaya çıkan esnekleşme süreci İş Hukukunda da farklı kavramlarla yeni düzenlemeleri gerektirmiştir. Bunlardan biri de denkleştirme uygulaması (yoğunlaştırılmış iş haftası/ sıkıştırılmış iş haftası)dır. Bireysel iş sözleşmeleri ile en çok iki ay, toplu iş sözleşmeleri ile en çok dört uygulanabilen denkleştirme, bazı haftalar haftalık çalışma süresinin aşılabildiği fakat günlük çalışma süresi olan 11 saatin aşılmadan diğer haftalarda ise haftalık çalışma süresinin altında bir çalışma süresi olacak şekilde düzenlenmektedir (Odaman, 2014:75). Burada önemli olan husus, günlük çalışma sınırı olan 11 saati ve ortalama haftalık çalışma süresinin de yasal sınırı aşmamasıdır. Diğer bir çalışma süresine göre esnek çalışma “telafi çalışması”dır. Zorunlu nedenlerle işin durması veya işçinin talebi üzerine yasal izin süreleri dışında izin verilmesi üzerine, çalışılmayan bu 184 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi sürenin en geç iki ay içinde yerine getirilmesini düzenleyen bir uygulamadır (Tezioğlu, 2021:170). Ekonomik kriz, zorlayıcı nedenler gibi durumlarda başvurulan diğer bir esnek istihdam türü de “kısa çalışma”dır. 4447 sayılı ek 2.md. göre “Genel ekonomik, sektörel veya bölgesel kriz ile zorlayıcı sebeplerle işyerindeki haftalık çalışma sürelerinin geçici olarak önemli ölçüde azaltılması veya işyerinin faaliyetinin tamamen veya kısmen durdurulması hallerinde, işyerinde üç ayı aşmamak üzere kısa çalışma yapılabilir”. İşletmelerin, özellikle Covid-19 pandemi sürecinde sıklıkla uyguladığı bir yöntem olmuştur. Böylece büyük ekonomik kriz veya zorlayıcı nedenlerin varlığı sürecinde işçinin iş sözleşmesini feshetmek yerine kısa çalışma talep etmek “feshin son çare” olması ilkesinin de uygulanmasında önemlidir. 1.2.2. İstihdam Biçimine Göre Esnek İstihdam Türleri İstihdam biçimine göre esnek istihdam türlerinden biri “evde çalışma”dır. Evde çalışma uygulamada üç şekilde görülmektedir. Bunlar; eve iş verme, ev hizmetlerinde çalışma ve evde bağımsız çalışmadır. Genel olarak evde çalışma sözleşmesi, ILO (Uluslararası Çalışma Örgütü) 177 Sayılı Evde Çalışma Sözleşmesi ve 184 sayılı Evde Çalışma Tavsiye Kararı çerçevesinde, “ulusal kanunlar, yönetmelik ve mahkeme kararları uyarınca bağımsız çalışma için gerekli olan özerklik ve bağımsızlığa sahip olmayan kişinin, kendi evinde veya işverenin işyeri dışında kendi seçtiği diğer yerde, bir ücret karşılığında malzeme, donanım ve kullanılan diğer girdilerin, kendisi, işveren veya aracı tarafından temin edilmesine bakılmaksızın, işveren tarafından belirlenen mal veya hizmetlerin üretimi için iş görmeyi üstlendiği sözleşmedir” (Şen, 2017: 479). Teknolojik gelişmeler; akıllı telefonların, tablet ve bilgisayarların yaygınlaşması, internet ağ sisteminin gelişmesi iş ve yaşam alanında önemli bir değişimi tetiklemiştir. Uzaktan çalışma kapsamında değerlendirilen bu esnek çalışma türü tele-çalışmadır. Bu uygulama ile çalışanların işleri esnetilmek istenmektedir. İş Kanunu uzaktan çalışmayı evden çalışma ve tele çalışmayı da kapsayacak şekilde düzenlemiştir. Buna göre “uzaktan çalışma; işçinin, işveren tarafından oluşturulan iş organizasyonu kapsamında iş görme edimini evinde ya da teknolojik iletişim araçları ile işyeri dışında yerine getirmesi esasına dayalı ve yazılı olarak kurulan iş ilişkisidir” (Ek fıkra: 6/5/2016-6715/2 md.). Tele çalışma ve evden çalışma uzaktan çalışmanın alt türü olarak değerlendirilmekle birlikte evden çalışma ve tele çalışma farklılık göstermektedir. Tele çalışma bilgi iletişim teknolojilerinden en az birini kullanarak merkez işyerinden uzakta yapılan bir çalışma iken evde çalışma 185 Özlem Demir iletişim teknolojilerinden birinin kullanılması ile gerçekleştirilebildiği gibi parça başı ücretin benimsendiği geleneksel çalışma biçimde de olabilmektedir (Erafşar, 2022:282). İstihdam biçimine göre esneklik kapsamında değerlendirilen diğer çalışma türü “alt işverenlik”tir. Son yıllarda istihdamın dışsallaştırılmasında sıkılıkla kullanılan alt işverenlik ilişkisi İş Kanunu’nda düzenlenmiştir: “Bir işverenden, işyerinde yürüttüğü mal veya hizmet üretimine ilişkin yardımcı işlerinde veya asıl işin bir bölümünde işletmenin ve işin gereği ile teknolojik nedenlerle uzmanlık gerektiren işlerde iş alan ve bu iş için görevlendirdiği işçilerini sadece bu işyerinde aldığı işte çalıştıran diğer işveren ile iş aldığı işveren arasında kurulan ilişkiye asıl işveren-alt işveren ilişkisi denir” (İş K. 2.md.). 4857 sayılı İş Kanunu’nda yer alan “geçici iş ilişkisi” esneklik kapsamında en fazla tartışılan konulardan bir olmuş, tasarıda “ödünç iş ilişkisi” olan kavram “geçici iş ilişkisi” olarak yasal düzenlemede yerini almıştır. Geçici iş ilişkisi, 2016 yılında yapılan bir değişiklikle (6715 sayılı Kanun) iki farklı uygulamaya ayrılmıştır. Birincisi “meslek edinilmemiş geçici iş ilişki”, ikinci ise “meslek edinilmiş geçici iş ilişkisi”dir. 2.1. Meslek Edinilmemiş Geçici İş İlişkisi Meslek edinilmemiş geçici iş ilişkisinde bir işveren yanında çalışmakta olan işçinin rızası alınarak geçici olarak başka bir işverene iş görme borcunu yerine getirmesi durumunu ifade etmektedir. Bu iş ilişkisinde ödünç veren işveren, ödünç alan işveren ve işçi arasında kurulan üçlü bir hukuki ilişki söz konusudur. Meslek edinilmemiş geçici iş ilişkisinde iş sözleşmesi ödünç veren işverenle devam etmektedir. Fakat sözleşmeden doğan iş görme borcu ödünç alan işverene devredilmekte yani işçi iş görme borcunu ödünç alan işverene karşı ifa etmekle yükümlü olacaktır. Buna bağlı olarak ödünç alan işveren emir ve talimat verme yetkisine de sahip olacaktır (Başbuğ, :2016107). 4847 sayılı İş Kanunu’nun 7.md. göre, “İşverenin, devir sırasında yazılı rızasını almak suretiyle bir işçisini, holding bünyesi içinde veya aynı şirketler topluluğuna bağlı başka bir işyerinde iş görme edimini yerine getirmek üzere geçici olarak devretmesi hâlinde de geçici iş ilişkisi kurulmuş olur. Bu fıkra kapsamında geçici iş ilişkisi, yazılı olarak altı ayı geçmemek üzere kurulabilir ve en fazla iki defa yenilenebilir” (İş K. md.7/15). Madde hükmünden açıkça anlaşıldığı gibi meslek edinilmemiş geçici iş ilişkisinde süre sınırı konulmuştur. Altı ayı geçmemek üzere yapılacağı ve en fazla da iki defa daha yenilenebileceği kabul edilmiştir. Bu hükme göre bu sürenin uzatılmasının mümkün olmadığı anlaşılmaktadır. İlgili maddenin devamında “İşçisini geçici olarak devreden işverenin ücret ödeme yükümlülüğü devam eder. Geçici iş ilişkisi kurulan işveren, işçinin kendisinde çalıştığı sürede ödenmeyen ücretinden, işçiyi gözetme borcundan ve sosyal sigorta primlerinden, devreden işveren ile birlikte sorumludur” ifadesi ile ödünç veren işverenin ücret ödeme yükümlülüğünün devam ettiği, ödünç alan işveren ise kendisinde çalıştığı süre içinde ödenmeyen ücretinden, sigorta primlerinden ödünç veren işverenle birlikte sorumlu tutulmuştur. 2. Geçici İş İlişkisi ve Türleri Bir firma esnek üretim ve esnek istihdam yöntemlerini kullanarak esnekliğe ulaşabilir. Bu nedenle son yıllarda işgücü piyasasında esnek istihdam biçimleri yaygınlık kazanmıştır. Geçici istihdam, bir “tampon” mekanizma olarak kullanıldığı için talep dalgalarına göre ayarlanmış esnek bir yapı sunmaktadır (Pfeifer, 2005:406). İşgücü ihtiyacı olan işletmeler, kadrolu ya da daimi çalışan almak yerine maksimum esneklik sağlayan geçici işgücü temin etme yolunu tercih etmektedirler. Böylece işletmeler, işgücünü genişletme kararı alırken, gelecekteki işten çıkarmaların bir sonucu olarak ortaya çıkabilecek maaliyetleri hesaba katmaktadırlar. Geçici istihdam ile işçilik ve genel idari giderler, eğitim, seçim ve personel alımı ile ilgili maliyetlerin azaltılması hedeflenmektedir. Ayrıca geçici istihdam ile hastalık, tatil ve daimi çalışanların diğer devamsızlıkları için bir “köprü” aracı olarak ve işe alım mekanizması olarak tercih edilmektedir. İş yükü dalgalanmaları ve işçi devamsızlıklarına uyum sağlama kapasitesinin artması gibi ihtiyaçlar nedeniyle esnek istihdam yaygın olarak kullanılmaktadır (Houseman, 2001; Foote ve Folta, 2002; Barbieri, Foote ve Folta, 2002:579). İşletmeler açısından işgücü maaliyetlerini sınırlama amacı taşıyan geçici iş ilişkisi, ekonomik durgunluk dönemlerinde işsizlik sorununa da bir çözüm olarak görülmüştür. Uygulamada ve yasal düzenlemede iki tür geçici iş ilişkisi söz Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi 186 konusudur; birincisi meslek edinilmemiş geçici iş ilişkisi, ikincisi ise meslek edinilmiş geçici iş ilişkisidir. 2.2. Meslek Edinilmiş Geçici İş İlişkisi Küresel ekonomik değişimler, işletmeler arasındaki rekabeti ve belirsizliği artırmıştır. Daha fazla kar elde etme ve maliyetleri düşürme baskısı, işletmeleri istihdam konusunda standart dışı farklı yaklaşımlara yönlendirmiştir. Bu yeni standart dışı istihdam türlerinden biri de “meslek edinilmiş geçici iş ilişkisidir”. Genel bir tanımlama ile meslek edilmiş geçici iş ilişkisi, “özel istihdam bürolarının kendisine iş sözleşmesi ile bağlı olan işçilerini işçi temini sözleşmeleri yoluyla geçici işverenlere devretmesiyle söz konusu olan geçici iş ilişkisini ifade etmektedir” (Baskan, 2017:3). Özel istihdam bürosu aracılığı 187 Özlem Demir Özlem Demir ile geçici iş ilişkisi, önemli bir istihdam kaynağı olma durumundadır. Özel istihdam bürolarından işçi talep etmek, işletmelerin istihdam fonksiyonunu veya işgücünün idari sorunluluğunu dışsallaştırmak üzere esneklik sağlama isteğinden kaynaklanmaktadır (Parlak ve Özdemir, 2011:20). 4857 sayılı İş Kanunu 7. md. yapılan değişiklikle (06.05.2016 tarih ve 6715 sayılı Kanun) Türk İş Hukuku’nda yeni bir uygulama olan “mesleki anlamda geçici iş ilişkisi” başka bir ifade ile “meslek edinilmiş geçici iş ilişkisi” düzenlenmiştir. Buna göre “Geçici iş ilişkisi, özel istihdam bürosu aracılığıyla ya da holding bünyesi içinde veya aynı şirketler topluluğuna bağlı başka bir işyerinde görevlendirme yapılmak suretiyle kurulabilir” (İş K. 7.md.). İlgili hükme göre geçici iş ilişkisi ya özel istihdam bürosu aracılığı ile kurulabilecek ya da holding bünyesi içinde veya aynı şirket topluluğuna bağlı başka bir işyerinde görevlendirmek suretiyle geçici iş ilişkisi kurulabilecektir. Buna bağlamda, özel istihdam bürosu aracılığı ile gerçekleşen geçici iş ilişkisi “meslek edinilmiş geçici iş ilişkisi” iken, holding bünyesi içinde veya aynı şirket topluluğuna bağlı başka bir işyerinde görevlendirmek suretiyle gerçekleşen geçici iş ilişkisi ise “meslek edinilmemiş geçici iş ilişkisi”sini ifade etmektedir. Meslek edinilmiş geçici iş ilişkisi kurulabilmesi için kanun hükmünde de açıkça belirtildiği gibi Özel İstihdam Bürolarının varlığı gereklidir. 2.2. Meslek Edinilmiş Geçici İş İlişkisi İlgili maddenin devamında bu düzenlemenin ayrıntıları yer almaktadır: “Özel istihdam bürosu aracılığıyla geçici iş ilişkisi, Türkiye İş Kurumunca izin verilen özel istihdam bürosunun bir işverenle geçici işçi sağlama sözleşmesi yaparak bir işçisini geçici olarak bu işverene devri ile; a) Bu Kanunun 13 üncü maddesinin beşinci fıkrası ile 74 üncü maddesinde belirtilen hâllerde, işçinin askerlik hizmeti hâlinde ve iş sözleşmesinin askıda kaldığı diğer hâllerde, b) Mevsimlik tarım işlerinde, c) Ev hizmetlerinde, d) İşletmenin günlük işlerinden sayılmayan ve aralıklı olarak gördürülen işlerde, e) İş sağlığı ve güvenliği bakımından acil olan işlerde veya üretimi önemli ölçüde etkileyen zorlayıcı nedenlerin ortaya çıkması hâlinde, f) İşletmenin ortalama mal ve hizmet üretim kapasitesinin geçici iş ilişkisi kurulmasını gerektirecek ölçüde ve öngörülemeyen şekilde artması hâlinde, g) Mevsimlik işler hariç dönemsellik arz eden iş artışları hâlinde, kurulabilir” (İş K. 7.md). Ayrıca kanun koyucu hangi durumlarda geçici iş ilişkisinin yapılabileceğini saymakla kalmamış süre açısından da bir sınırlama getirmiştir. Bu hükme göre, “Geçici işçi sağlama sözleşmesi ikinci fıkranın (a) bendinde sayılan hâllerin devamı süresince, (b) ve (c) bentlerinde sayılan hâllerde süre sınırı olmaksızın, diğer bentlerde sayılan hâllerde ise en fazla dört ay süreyle kurulabilir. Yapılan bu sözleşme ikinci fıkranın (g) bendi hariç toplam sekiz ayı geçmemek üzere en fazla iki defa yenilenebilir. Geçici işçi çalıştıran işveren, belirtilen sürenin 188 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi sonunda aynı iş için altı ay geçmedikçe yeniden geçici işçi çalıştıramaz” (İş K. 7.md). (İş Özel istihdam bürosu aracılığı ile “meslek edinilmiş geçici iş ilişkisi”nin kurulamayacağı durumlarda düzenlemiştir. “Bu Kanunun 29 uncu maddesi kapsamında toplu işçi çıkarılan işyerlerinde sekiz ay süresince, kamu kurum ve kuruluşlarında ve yer altında maden çıkarılan işyerlerinde bu maddenin ikinci fıkrası kapsamında geçici iş ilişkisi kurulamaz. Geçici işçi çalıştıran işveren, grev ve lokavtın uygulanması sırasında 18/10/2012 tarihli ve 6356 sayılı Sendikalar ve Toplu İş Sözleşmesi Kanununun 65 inci maddesi hükümleri saklı kalmak kaydıyla geçici iş ilişkisiyle işçi çalıştıramaz” (İş K. 7.md) hükmü ile geçici iş ilişkisinin kurulamayacağı durumlar, dört kapsamda değerlendirilmiştir. Geçici iş ilişkisinde, geçici işçinin işvereninin özel istihdam bürosu olduğu kanun maddesinde açıkça belirtilmiştir. Buna göre “Geçici iş ilişkisinde işveren özel istihdam bürosudur. Özel istihdam bürosu aracılığıyla geçici iş ilişkisi, geçici işçi ile iş sözleşmesi, geçici işçi çalıştıran işveren ile geçici işçi sağlama sözleşmesi yapmak suretiyle yazılı olarak kurulur. 2 Özel İstihdam Bürosunun uygulama esaslarını belirlemek amacıyla 11.10.2016 tarih ve 29854 sayılı “Özel İstihdam Büroları Yönetmeliği” Resmi Gazetede yayınlanarak yürürlüğe girmiştir. İlgili yönetmelik kapsamında kurumun kurulması, faaliyetleri, denetimi, kontrollerine ilişkin esaslar belirlenmiş, meslek edinilmiş geçici iş ilişkisinin detaylarına yer verilmiştir. 2.2. Meslek Edinilmiş Geçici İş İlişkisi Özel istihdam bürosu ile geçici işçi çalıştıran işveren arasında yapılacak geçici işçi sağlama sözleşmesinde; sözleşmenin başlangıç ve bitiş tarihi, işin niteliği, özel istihdam bürosunun hizmet bedeli, varsa geçici işçi çalıştıran işverenin ve özel istihdam bürosunun özel yükümlülükleri yer alır” (İş K. 7.md). Özel istihdam bürosu işveren konumunda geçici iş ilişkisinin tarafıdır. Bu durumda özel istihdam bürosu, işçi ve geçici işverenin olduğu üç taraflı bir istihdam ilişkisi söz konusudur. Özel istihdam bürosu ile işçi arasında bir iş sözleşmesi yapılırken, özel istihdam bürosu ile geçici işveren arasında da “geçici işçi sağlama sözleşmesi” yapılmaktadır2. Türkiye İş Kurumu tarafından izin verilen ve faaliyetleri devam eden (aktif) toplam 695 özel istihdam bürosu bulunmaktadır. Türkiye İş Kurumu tarafından izin verilen ve geçici iş ilişkisi yapma yetkisi alan özel istihdam büro sayısı ise 37’dir. Bu bürolar başta İstanbul olmak üzere Türkiye’nin büyük şehirlerinde hizmet vermektedir (İşkur, 2023). 189 Özlem Demir 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi Esnek çalışma modellerinin işgücü piyasasına etkileri; istihdam ve işsizlik üzerine etkisi, bireysel iş ilişkileri, toplu iş ilişkisi, sosyal güvenlik ve iş sağlığı ve güvenliği gibi temel alanlarda söz konusu olmaktadır. Çalışmanın konusu olan meslek edinilmiş geçici iş ilişkisinde işçiye sağlanan hakların, İş Hukuku’nun temel esasları çerçevesinde yeniden gözden geçirilmesi gerekliliği doğmuştur. Meslek edinilmiş geçici iş ilişkisi yakın zamanda yapılan düzenleme ile yasal zemini oluşturulmuştur. Ancak bazı ihtilaflı durumlar söz konudur. Bu sorunların yanı sıra iş tatmini, iş güvencesi, kariyer gelişimi gibi çalışma yaşamına dair diğer sorunları da içeren bir istihdam biçimi olması bakımından önemlidir. Genel olarak esnek istihdam biçimlerinin istihdam ve işsizlik üzerine etkileri tartışmalıdır. Kimi görüşe göre esnek uygulamalar işsizliği azaltmakta, kimi görüşe göre ise istihdam ve işsizlik üzerine doğrudan bağlantı bulunmamaktadır. Esneklik ile istihdam ve işsizlik arasında doğrudan bağlantının bulunmaması büyük ölçüde esnekliğin istihdamı etkileyen faktörlerden sadece bir olması ile ilgilidir. Ancak esnek istihdam biçimlerinin istihdam yapısını doğrudan etkilediği bir gerçektir. Evde çalışma, kısmi süreli çalışma, tele çalışma, geçici iş ilişkisi gibi standart dışı istihdam biçimleri, kadınlar, gençler, yaşlılar, göçmenler, engelli bireylerin çalışma hayatına girmelerine olanak tanımış, iş deneyim fırsatı sağlamıştır (Tokol, 2017:222). Meslek edinilmiş geçici iş ilişkisi, kalıcı istihdam içindeki payları düşük olan bu gruplara iş ve yaşam dengesi sunması nedeniyle tercihe konu olmakta istihdam edilebilirliği artırmaktadır. Ayrıca geçici iş ilişkisinin işçilere, işverenler arasında sık sık değişiklik yaparak becerilerini geliştirme ve mesleki bilgi seviyelerini artırma olanağı sağladığı da vurgulanmaktadır (Ritter- Hayashi,Knoben ve Vermeulen, 2021:). Geçici iş ilişkisi ile çalışan işçilerin, aynı işte uzun süreli çalışmaması, önyargılı insan kaynakları politikaları, işçiler arasında bölünme ve güvencesiz iş yaşamından olumsuz etkilendikleri bilinmektedir. Ayrıca eğitimden, beceriden, kariyer gelişiminden daimi çalışanlara göre daha fazla mahrum kalmaktadırlar. Bu durum işyerine bağlığı ve iş tatminini çoğunlukla olumsuz etkilediği belirtilmektedir (Manelkar, 2023:267). İşgücü piyasasına ilişkin geçici iş ilişkisinin bahsi geçen farklı sonuçlarının yanı sıra İş Hukuku alanına dair ihtilaflı konuları da tartışma konusu olmaktadır. Bireysel iş hukuku bakımından emir ve talimat verme hakkı, ücret, iş sözleşmesinin devamı, işçiye eşit davranma borcu, işçinin sadakat borcu gibi hususlar ön plana çıkmaktadır. Geçici iş ilişkisinde “yönetim hakkı”nın (emir 190 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi ve talimat verme yetkisi) hangi işverende olduğunun belirlenmesi önemlidir. Kural olarak yönetim hakkı geçici işverene geçtiği kabul edilmekle birlikte özel istihdam bürosunun belli ölçütlerde emir ve talimat verme hakkı vardır (Baskan, 2017:26). Geçici iş ilişkisinde diğer bir sorunlu husus ise “ücret” konusudur. 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi Genel kural ücreti, iş sözleşmesinin tarafı olan özel istihdam bürosunun ödeyeceğidir. Ancak çalışmadığı sürece ücret ve sosyal güvenlik prim ödemesi yapılmamaktadır. Bu süresinin uzaması durumunda işçiye iş sözleşmesini feshetme imkanı tanınmıştır (Günaydın ve Özdemir, 2018:12). Şöyle ki “Geçici işçi ile yapılacak iş sözleşmesinde, işçinin ne kadar süre içerisinde işe çağrılmazsa haklı nedenle iş sözleşmesini feshedebileceği belirtilir. Bu süre üç ayı geçemez” (İş K. 7.md.) hükmü ile yasal süre sınırı belirlenmiştir. İş sözleşmesinin devamı anlamında belirli süreli iş sözleşmesi ya da belirsiz süreli iş sözleşmesi olup olmadığı da ayrı bir sorundur. Bu düzenleme açıkça yer almadığından sözleşmenin türü kurulacak iş ilişkisinin özelliğine göre belirlenecektir. Başka bir ifade ile iş sözleşmesinin belirli, belirsiz, tam süreli, kısmi süreli, evde çalışma, tele çalışma sözleşmesi olması işin türü ve somut durum belirleyici olacaktır. Özel istihdam bürosu ile geçici işçi arasında kurulacak olan sözleşmenin belirli süreli olması ancak İş Kanunun 11. maddesinde yer alan objektif koşulların varlığı ile mümkün olacağı açıktır (Civan, 2017:318-318). Eşit davranma borcu, meslek edinilmiş geçici iş ilişkisinde önemli düzenlemelerden biridir. İş Kanunu’nda açıkça belirtildiği üzere “Geçici işçinin, geçici işçiyi çalıştıran işverenin işyerindeki çalışma süresince temel çalışma koşulları, bu işçilerin aynı işveren tarafından aynı iş için doğrudan istihdamı hâlinde sağlanacak koşulların altında olamaz” (İş K. 7. md). Bu maddede ifade edilen temel çalışma koşulları; genel tatiller, ücretli izin, ara dinlenme süreleri, gece çalışması, fazla çalışma gibi durumları içermektedir. Ayrıca cinsiyet eşitliği, ırk, etnik köken, din, inanç gibi unsurlar ayrımcılık yapmama kapsamında temel çalışma koşulları çerçevesinde değerlendirilmektedir (Akpınar, 2021:129). Geçici çalışan işçi, ödünç alan işverenin çalışma düzenine diğer çalışan işçiler gibi uymak durumunda ve ödünç alan işverene karşı sadakat borcu bulunmaktadır. Ayrıca meslek edinilmiş geçici iş ilişkisi, üçlü bir hukuki ilişki olduğundan geçici işçinin ödünç alan işverene olduğu gibi özel istihdam bürosuna karşı da sadakat borcu bulunmaktadır (Civan, 2017:333). ve talimat verme yetkisi) hangi işverende olduğunun belirlenmesi önemlidir. Kural olarak yönetim hakkı geçici işverene geçtiği kabul edilmekle birlikte özel istihdam bürosunun belli ölçütlerde emir ve talimat verme hakkı vardır (Baskan, 2017:26). Geçici iş ilişkisinde diğer bir sorunlu husus ise “ücret” konusudur. Genel kural ücreti, iş sözleşmesinin tarafı olan özel istihdam bürosunun ödeyeceğidir. Ancak çalışmadığı sürece ücret ve sosyal güvenlik prim ödemesi yapılmamaktadır. Bu süresinin uzaması durumunda işçiye iş sözleşmesini feshetme imkanı tanınmıştır (Günaydın ve Özdemir, 2018:12). Şöyle ki “Geçici işçi ile yapılacak iş sözleşmesinde, işçinin ne kadar süre içerisinde işe çağrılmazsa haklı nedenle iş sözleşmesini feshedebileceği belirtilir. Bu süre üç ayı geçemez” (İş K. 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi 7.md.) hükmü ile yasal süre sınırı belirlenmiştir. İş sözleşmesinin devamı anlamında belirli süreli iş sözleşmesi ya da belirsiz süreli iş sözleşmesi olup olmadığı da ayrı bir sorundur. Bu düzenleme açıkça yer almadığından sözleşmenin türü kurulacak iş ilişkisinin özelliğine göre belirlenecektir. Başka bir ifade ile iş sözleşmesinin belirli, belirsiz, tam süreli, kısmi süreli, evde çalışma, tele çalışma sözleşmesi olması işin türü ve somut durum belirleyici olacaktır. Özel istihdam bürosu ile geçici işçi arasında kurulacak olan sözleşmenin belirli süreli olması ancak İş Kanunun 11. maddesinde yer alan objektif koşulların varlığı ile mümkün olacağı açıktır (Civan, 2017:318-318). Eşit davranma borcu, meslek edinilmiş geçici iş ilişkisinde önemli düzenlemelerden biridir. İş Kanunu’nda açıkça belirtildiği üzere “Geçici işçinin, geçici işçiyi çalıştıran işverenin işyerindeki çalışma süresince temel çalışma koşulları, bu işçilerin aynı işveren tarafından aynı iş için doğrudan istihdamı hâlinde sağlanacak koşulların altında olamaz” (İş K. 7. md). Bu maddede ifade edilen temel çalışma koşulları; genel tatiller, ücretli izin, ara dinlenme süreleri, gece çalışması, fazla çalışma gibi durumları içermektedir. Ayrıca cinsiyet eşitliği, ırk, etnik köken, din, inanç gibi unsurlar ayrımcılık yapmama kapsamında temel çalışma koşulları çerçevesinde değerlendirilmektedir (Akpınar, 2021:129). Geçici çalışan işçi, ödünç alan işverenin çalışma düzenine diğer çalışan işçiler gibi uymak durumunda ve ödünç alan işverene karşı sadakat borcu bulunmaktadır. Ayrıca meslek edinilmiş geçici iş ilişkisi, üçlü bir hukuki ilişki olduğundan geçici işçinin ödünç alan işverene olduğu gibi özel istihdam bürosuna karşı da sadakat borcu bulunmaktadır (Civan, 2017:333). Meslek edinilmiş geçici iş ilişkisinin toplu iş hukuku açısından etki ve sonuçları değerlendirildiğinde; sendika üyeliği, işkolunun tespiti, toplu iş sözleşmesinden yararlanma ve grev-lokavt durumu gibi unsurlar öne çıkmaktadır. Öncelikle bu kapsamda geçici işçinin özel istihdam bürosunun girdiği işkolunda mı yoksa geçici olarak iş görme edimini yerine getirdiği işyerinin girdiği işkolunda mı sendikaya üye olacağının belirlenmesi 191 Özlem Demir Özlem Demir önemlidir. Sendika üyeliği “6356 Sayılı Sendikalar ve Toplu İş Sözleşmesi Kanunu”na göre işkoluna göre olmaktadır. Bu nedenle işkolunun tespiti önemli bir unsurdur. Geçici işçi, iş görme borcunu ödünç alan işverenin işyerinde ifa etmesine rağmen asıl işvereni özel istihdam bürosudur. Bu bağlamda geçici işçinin sendika üyeliği, özel istihdam bürosunun girdiği işkolu esas alınmaktadır. Ayrıca geçici işçi özel istihdam bürosunun taraf olduğu toplu iş sözleşmesinden yararlanabilecektir (Civan, 2017:384). Toplu iş hukukunun konusu olan başka bir husus ise greve katılma durumudur. Geçici işçinin hangi işverenin işyerindeki greve katılacağının tespiti önemlidir. 3. Meslek Edinilmiş Geçici İş İlişkisinin İşgücü Piyasası Açısından Değerlendirilmesi Doktrinde faklı görüşler bulunmakla birlikte özel istihdam bürosunun işyerinde yapılacak olan bir greve geçici işverenin yanında çalışan özel istihdam bürosunun işçisinin katılabileceğinin benimsenmesi hem kanun hem de Anayasal düzenlemeye uygun olacaktır. Böylelikle, geçici işçi geçici işverenin (ödünç alan işverenin) işyerindeki iş görme borcunu yerine getirmeyerek özel istihdam bürosundaki greve katılmış olacaktır (Civan, 2017:387, Akpınar, 2021:168). Sosyal güvenlik açısından işçinin geçici olarak başka bir işverene verilerek çalıştığı durumda ve başka bir işverenin yanında geçici olarak çalışmadığı durumdaki sigortalılığının ayrı değerlendirilmesi gerekir. İşçinin geçici olarak başka bir işverene verilerek çalıştığı durumda işçi, özel istihdam bürosu tarafından başka işverene ödünç verilmek üzere alınmıştır. Başka bir ifade ile işçinin işvereni, özel istihdam bürosudur ve özel istihdam bürosuna ait işyerinde fiilen çalışmaması sigortalı olarak kabul edilmesi konusunda herhangi bir engel içermez. İşçinin başka bir işverenin yanında geçici olarak çalışmadığı durumda ise sigorta primlerinin devam ettirilmesi gerektiği veya çalışmadığı için sigorta primlerinin yatırılmasının söz konusu olmadığı gibi öğretide farklı görüşler vardır. Bu nedenle kanun koyucunun işçinin çalışmadığı sürece ücret ve sigorta primi ödeme yükümlülüğüne ilişkin ayrı bir düzenleme yapması gerekliliği savunulmaktadır (Öktem-Songu ve Şahin- Emir, 2020:239-241). İş sağlığı ve güvenliği kapsamında da geçici iş ilişkisine özgü sorunlar bulunmaktadır. İş kazası ve meslek hastalığını bildirme yükümlülüğünün hangi işverende olduğu, geçici çalışan işçinin uğradığı iş kazasındaki sorumluluğun tespit edilmesi önemlidir. Geçici işçi çalıştıran işveren “Geçici işçinin iş kazası ve meslek hastalığı hâllerini özel istihdam bürosuna derhâl, 31/5/2006 tarihli ve 5510 sayılı Sosyal Sigortalar ve Genel Sağlık Sigortası Kanunu’nun 13 üncü ve 14 üncü maddelerine göre ilgili mercilere bildirmekle yükümlü” (İş K. 7.md) tutulmuştur. İş kazası ve meslek hastalığını bildirme yükümlülüğü geçici işçi çalıştıran işveren üzerinde olmakla birlikte iş kazası ve meslek hastalığından doğan zararın geçici işçi çalıştıran işverende mi 192 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi yoksa özel istihdam bürosunda mı olacağı ilgili kanunda düzenlenmemiştir. Geçici iş ilişkisinde işçiyi koruma ve gözetme borcu her iki işverende de olduğundan iş sağlığı ve güvenliği önlemlerini almayan, eksik alan ve mevzuat hükümlerini yerine getirmeyen geçici işçi çalıştıran işveren, geçici işçinin iş kazası ve meslek hastalığına uğraması durumunda özel istihdam bürosu ile birlikte sorumlu tutulacaktır (Öktem-Songu ve Şahin-Emir, 2020:248). Ayrıca 6331 sayılı Kanunda “çalışanların eğitim” başlığı taşıyan 17.madde de “Geçici iş ilişkisi kurulan işveren, iş sağlığı ve güvenliği risklerine karşı çalışana gerekli eğitimin verilmesini sağlar (6631 Sayılı İş Sağlığı ve Güvenliği Kanunu 17.md.)” hükmü ile eğitim verme yükümlülüğü geçici işçi çalıştıran işverendedir. Sonuç Küreselleşme, bilgi teknolojilerindeki hızlı ilerleme, artan uluslararası rekabet üretim ve istihdam biçimlerinde önemli değişikliklere yol açmıştır. Post-fordist üretim sisteminden günümüze kadar olan bu değişiklik, farklı boyutlarda ve türlerde meydana gelse de “esneklik” paydasında şekillenmiştir. Bu süreç özellikle 1970’lerden sonra çalışma sistemlerinin ve endüstri ilişkilerinde esneklik aramaya yönlendiren koşulları artırmıştır. Katı mevzuat hükümleri de bu süreçte esnetilmiş, çalışma süreleri ve çalışma biçimleri bakımından farklı istihdam biçimleri yasal zemine kavuşmuştur. Bu esnek çalışma biçiminden biri de geçici iş ilişkisi olup, iki tür olarak düzenlenmiştir. Birincisi “meslek edinilmemiş geçici iş ilişkisi” iken, ikincisi “meslek edinilmiş geçici iş ilişkisi”dir. Son yıllarda işgücü piyasasında esnek işlerde önemli artış yaşanmıştır. Dünya genelinde yaşanan istihdam azalışına bağlı olarak esnek istihdam modellerinin işsizliği önlemede de bir çözüm aracı olarak görülmüştür. Standart dışı istihdam ciddi endişelere de yol açmaktadır; iş ve gelir güvencesi eksikliği, kariyer gelişimi zorlukları, sosyal yardım eksikliği, ücret ve ücrete bağlı haklarda eksiklik, iş stresi, yasal hak kayıpları, iş yaşam dengesi zorlukları, işyerinde ayrımcılık, sömürü ve sosyal dışlanma gibi riskleri artıran bir durum olarak görülmektedir. İşgücü piyasası açısından esnek çalışma biçimlerinin yaygınlaşmasının getireceği sorunlar; kayıt dışı çalışmanın artması, iş güvencesi ve gelir güvencesindeki yoksunluk, iş sağlığı ve güvenliği tehditleri, sendikal örgütlenme ve sendikal haklardan yararlanma sorunu, sosyal hak kayıpları öne çıkmaktadır. Geçici istihdama yönelik sorunların çözümünde yasal düzenlemeler, geçici işçilerin hak ve menfaatlerinin güvence altına alınması 193 Özlem Demir konusunda alt yapı oluşturmaktadır. Ancak işçiler açısından yeterli düzeyde koruyucu düzenlemelerin olup olmadığı tartışma konusudur. Meslek edinilmiş geçici iş ilişkisi özel istihdam bürosu aracılığı ile kurulmakta olup, iş ilişkileri açısından hukuki sonuçlar doğurmaktadır. Bu kapsamda bireysel iş ilişkileri, toplu iş ilişkileri, sosyal güvenlik ve iş sağlığı ve güvenliği alanında doğacak yükümlülükler ve sonuçların iyi değerlendirilmesi hak kayıplarının önlenmesi anlamında önemlidir. Güvenceli ve sağlıklı bir çalışma, çalışma isteğinde olan herkes için temel bir insan hakkıdır. Bu nedenle esnekliğin en önemli bileşeninin istihdam güvencesi, gelir güvencesi ve sosyal güvence olmalı, esneklik “güvenceli esneklik” anlayışı ile uygulanmalıdır. 194 Bir Esnek İstihdam Biçimi Olarak Geçici İş İlişkisi Kaynakça Akpınar, F. (2021). Mesleki Anlamda Geçici İş İlişkisi, Taşhan Kitap Yayınları, Tokat. Barbieri, P. (2009). Flexible Employment and Inequality in Europe. European Sociological Review, 25 (6), 621-628. Baskan, Ş. E. (2017). Türk İş Hukukunda Meslek Edinilmiş Geçici İş İlişki- si. 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Bölüm 12 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi Fazilet Nohut1 Özlem Balaban2 Özet Değişimin hiç olmadığı kadar hızlı rekabetin ise hiç olmadığı kadar acımasız olduğu bir çevrede örgütlerin ayakta kalmalarının en önemli belirleyicisi doğru stratejik kararların alınmasıdır. Ancak doğru stratejik kararların alınması sürdürülebilir rekabet avantajının elde edilmesi konusunda tek başına yeterli değildir. Örgütün stratejik kararlarının örgütün tüm fonksiyonlarıyla uyumlu olması, bütüncül bir bakış açısıyla aynı örgütsel amaçları gerçekleştirmeye yönelmesi gerekmektedir. Örgütlerde strateji yapıcılar ve uygulayıcılar insanlardan oluşmaktadır. Dolayısıyla insanın kendisi başlı başına örgüte rekabet üstünlüğü kazandırabilecek bir stratejik unsur olarak ele alınmalıdır. Kaynak temelli yaklaşıma göre ikamesi ve taklit edilmesi zor olan insan kaynağının örgütün temel stratejik amaçlarıyla uyumlu bir şekilde yönlendirilmesi son derece önemlidir. Bu çalışmada genel işletme stratejileri ve insan kaynakları yönetimi uygulamalarının yatay ve dikey uyum süreçleriyle bütünleşmesinin örgütün genel performansı üzerinde nasıl bir etkiye sahip olduğu açıklanmaya çalışılmıştır. Genel işletme stratejilerinin çerçevesi ise Porter’ın önerdiği jenerik (rekabetçi stratejiler) stratejilerle sınırlandırılmıştır. Yatay ve dikey uyum sağlama süreçlerinde yöneticilere düşen roller, bu süreçlerde karşılaşılabilecek sorunlar ve örgüt yapısının önemi de bu çalışmada ele alınmıştır. Özellikle insan kaynakları süreçlerinin yapılandırılmasında, stratejik kararların alınması ve uygulanmasında örgüt yapılarının da stratejik bir role sahip olduğu düşünüldüğünde mekanik ve organik örgüt yapılarının açıklanmasına yer verilmiştir. Ve bu örgüt yapılarının insan kaynakları süreçlerinin tasarımında, uyum ve bütünleşme sürecine katkılarının stratejik önemi de vurgulanmıştır. Sonuç olarak bu çalışmayla stratejik insan kaynakları yönetiminin genel işletme performansı üzerindeki olumlu etkilerinin altını çizerek alana katkı sunmak amaçlanmıştır. https://doi.org/10.58830/ozgur.pub401.c1760 https://doi.org/10.58830/ozgur.pub401.c1760 197 \| Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi 198 GİRİŞ GİRİŞ İşletmelerde “rekabet” kavramının kapsayıcılığının artmasıyla beraber örgütler buna yönelik aksiyonlar almaya başlamışlardır. Bununla beraber işletmelerde bir iş stratejisi benimsemek zorunluluk halini almıştır. Bu strateji işletmeyi rakiplerinden farklılaştıran ve onun rekabet üstünlüğü sağlamasına olanak tanıyan bir takım uygulamaları beraberinde getirmiştir. İşletmeler varlıklarını sürdürürken her zaman aynı uygulamaları devam ettirmemekte çevrede meydana gelen değişikliklere göre farklı stratejiler uygulamaktadırlar. İşletmelerin bu gelişim ve değişim süreçlerinde çevresel uyarıların farkına varıp bunlara tepki ve uyum sağlama olanakları olabilmektedir. Bu uyum işletmenin benimsediği stratejik tercihlere göre şekillenmektedir. Bu çalışmada işletmelerde yazında en yaygın ifade edilen genel işletme stratejilerine değinilmektedir. Bunlardan yazında en sık ifade edilen rekabet stratejilerinden maliyet liderliğinde sıkı finansal kontrol ile daha uygun iş gücü ihtiyacı ve daha az esnek bir örgütsel yapı varken farklılaştırma stratejisinde daha esnek yapı ve denetimle yetkinlik temelli insan kaynağı ihtiyacı söz konusudur. Odaklanma da ise bir ürüne veya belli bir müşteri kitlesi veya coğrafyaya odaklanarak örgütsel yapıyı buna göre şekillendirme söz konusudur. Bu ve buna benzer stratejiler hem örgüt içinde hem de örgütün dış çevreye olan yanıtına karar vermede bağlayıcı unsurlar olmaktadır. İşletmelerde “rekabet” kavramının kapsayıcılığının artmasıyla beraber örgütler buna yönelik aksiyonlar almaya başlamışlardır. Bununla beraber işletmelerde bir iş stratejisi benimsemek zorunluluk halini almıştır. Bu strateji işletmeyi rakiplerinden farklılaştıran ve onun rekabet üstünlüğü sağlamasına olanak tanıyan bir takım uygulamaları beraberinde getirmiştir. İ Bir örgütün belirlediği strateji temel işletme fonksiyonlarıyla uyumlu olduğu zaman sürdürülebilir rekabet avantajı elde edilmesi mümkün olmaktadır. Dolayısıyla örgüt stratejilerinin oluşturmasında İnsan Kaynakları Yönetimi (IKY) fonksiyonu da mutlaka göz önünde bulundurulmalıdır. İKY’ nin karar ve uygulamalarında kendi fonksiyonları arasındaki uyum ve aynı zamanda işletmenin genel stratejisi ve diğer işletme fonksiyonlarıyla uyumu bütünleşik bir strateji anlayışını doğurmakta ve yüksek performans elde edilmesini sağlamaktadır. Bu bütünleşik anlayış içinde fonksiyonel bazda düşünüldüğünde Stratejik İnsan Kaynakları Yönetimi (SİKY) ortaya çıkmaktadır. Rakiplerinden daha akıllı ve esnek şirket (Boxall, 1996) yapısına sahip olmak günümüz dünyasının en önemli rekabet avantajı ya da bir diğer ifadeyle güçlü bir stratejik duruş olarak karşımıza çıkmaktadır. Yazında yer alan stratejik IK yönetimi ile ilgili yapılan tüm tanımlamaların ortak paydasının IK politika ve uygulamalarıyla sürdürülebilir bir rekabet avantajı elde etmek olduğu görülmektedir. Stratejik insan kaynakları yönetimi, insan kaynakları stratejilerinin kurumsal etkinliğe yaptığı katkıları ve bu katkıların nasıl ve hangi araçlarla elde edildiğinin anlaşılması (Ericksen ve Dyer, 2005) 199 Fazilet Nohut / Özlem Balaban örgütün genel stratejileriyle bütünleşme sürecinde oldukça önemlidir. GİRİŞ Örgütsel stratejilerin belirlenmesinde rolü ve önemi her geçen gün artan insan kaynakları yöneticilerinin örgütün faaliyet gösterdiği bağlam, örgütün iç çevresi, stratejisi, vizyonu, misyonu, politikaları, örgütün dış çevresi, rakipleri, müşterileri ve faaliyet gösterdiği toplumdaki sosyal, ekonomik ve yasal gelişmelere hakim olması son derece önemlidir. Ancak bu şekilde sağlam temeller üzerine inşaa edilmiş IK stratejilerinin örgütün genel performansı üzerinde olumlu bir etki meydana getirmesi beklenebilir. İnsan kaynakları yönetimindeki paradigma değişimiyle birlikte insan sermayesinin değerli, ikame ve taklit edilmesi güç ve kıt bir kaynak olmasının anlaşılması ve işletmelere önemli bir rekabet üstünlüğü sağlaması örgütlerin insan kaynakları politika, amaç ve uygulamalarının örgütsel amaç ve hedeflerle uyumlaştırılmasını zorunlu hale getirmiştir. Yukarıdaki gerekçelerden yola çıkarak bu çalışmada işletme stratejisi ile İKY stratejilerinin nasıl uyumlaştırılacağına ilişkin kavramsal açıklamalarda bulunulmaktadır. Bu temelde ilk olarak strateji kavramının teorik çerçevesi çizilmiş, daha sonra stratejik İKY’ye ilişkin kavramsal açıklamalar yapılmıştır. Akabinde İKY’de yatay ve dikey uyum kavramları açıklanmıştır. Uyum ve bütünleşme sürecinde IK profesyonellerinin rolü ve önemi, bu süreçte dikkat edilmesi gereken hususlara değinilmiş ve ardından rekabet stratejileri, örgüt yapıları ve IKY arasındaki stratejik uyum süreçleri açıklanmaya çalışılmıştır. 1. Stratejik Yönetim Kavramsal Çerçeve Her işletme, özellikle en büyük rakipleriyle kıyaslandığında daha üstün ya da daha zayıf kaynak ve yeteneklere sahiptir. İşletmeler üstünlük ve zayıflıklarıyla birlikte dış çevreden gelen fırsat ve tehditlere karşı mücadele eder, ayakta kalmaya çalışır. Yani bir diğer ifadeyle sahip olduğu kaynak ve yetenekleri rekabet üstünlüğü elde edebilmek adına en uygun kombinasyonlarla bir araya getirerek uzun vadeli başarılar elde etmeyi amaçlar. İşte bu noktada bir işletme için strateji kavramının hayati önemi ortaya çıkmaktadır(Dinçer, 2004). Strateji en yalın ifadesiyle “fark yaratma” ve “aksiyon alma” sanatıdır. İşletmenin sahip olduğu kaynakları rakiplerinden daha iyi kullanabilme veya rekabet üstünlüğü sağlayacak becerilere dönüştürme yoluyla farklılaşması bu sayede öz yetkinlik ve temel yeteneklerini inşaa etme süreci olarak da tanımlanabilir (Coşkun, 2016:104). Bu açıdan bakıldığında strateji bir “ezber bozma” oyunu olarak da değerlendirilebilir. Dolayısıyla strateji kavramına bakıldığında bir dinamizmin söz konusun olduğu görülmektedir. 200 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi İlgili yazın incelendiğinde strateji kavramıyla ilgili yapılan birçok tanımın olduğu görülmektedir. Minztberg strateji kavramını projektif bir yaklaşımla ele alarak geçmiş uygulamaların sonuçları ışığında geleceğe dönük yapılan planlar olarak değerlendirmektedir. Böylelikle geçmişteki deneyimlerin yeni stratejilerin tasarlanmasında rekabet üstünlüğünü sağlayacağını vurgulamaktadır (Sheppeck ve Militello, 2000: 5). Stratejik yönetim bir örgütün amaçlarına ulaşması için tasarlanan planların oluşturulması ve uygulanmasıyla sonuçlanan kararlar ve eylemler olarak tanımlanabilir(Pearce ve Robinson, 2015:3). 1.1. Stratejik Yönetimin Seviyeleri Stratejik yönetimin etkin bir şekilde gerçekleştirilebilmesi için strateji ekibinin örgütün üç düzeyinden (firma, iş ve işlevsel) karar alıcıların katılımıyla oluşması gerekmektedir. Bunun yanı sıra strateji ekibine şirket hakkında veri sağlayacak ve strateji ekibinin aldığı kararları uygulamaya geçirecek bir ekibe daha ihtiyaç vardır. Bu ekip ise orta ve alt düzey hat yöneticileri ve planlama ekiplerinden oluşmalıdır (Pearce ve Robinson, 2015:9). Buradan yola çıkarak stratejik yönetimin üç düzeyini aşağıdaki gibi açıklanmıştır (Coşkun, 2015; Dinçer, 2004). Fonksiyonel Strateji: Departmanların birbirleriyle bütünleşik bir şekilde işletme stratejisine katkıda bulunmasını ifade eder. Bu strateji işletmenin ana fonksiyonları olan üretim, pazarlama, finansman ve insan kaynakları gibi bölümlere ait kararlardan meydana gelir. Bu kararlara taktik adı da verilebilir. Bu aşamadaki stratejik kararlara daha çok kaynakların verimli kullanılmasına yönelik kararlardır. Bu seviyedeki stratejiler daha çok bir fonksiyon içindeki faaliyetlerin koordinasyonunu sağlar ve uygulamaya yönelik kararları içerir. Rekabet üstünlüğü ve sinerji bu seviyedeki iki önemli strateji öğesi olarak öne çıkmaktadır. İşletme Stratejisi: İşletme ve iş birimi düzeyinde özgün hedeflerin tespit edilmesini ifade eder. Bir işletme ile çevresi arasındaki stratejik uyumu sağlayan kararlardır. Bir işletme içindeki bölümlerle ilgili kararların koordinasyonu bu düzeydeki strateji ile gerçekleştirilir. Bu düzeyde daha çok rekabet sorunlarıyla ilgilenildiği için işletme seviyesinde kaynak dağılımı, karşılaştırmalı üstünlükler ve sinerji konuları ön plana çıkar. Bu seviyede alınan stratejik kararlar işletme bölümlerine ait politika ve kararlara da rehberlik eder. Şirket Stratejisi: Hangi endüstride hangi ürün ve hizmetlerin yer alacağının belirlenmesi, büyüme, çeşitlendirme, işbirliği vb. stratejilerin belirlenmesini ifade eder. Bir diğer anlamda farklı işletmeler ve çeşitlendirilmiş 201 Fazilet Nohut / Özlem Balaban iş gruplarına ve yapıya sahip bir şirketin bir bütün olarak stratejisi demektir. Bu düzeydeki strateji şirketin bünyesindeki tüm işletmeleri kapsadığı için bir bütün olarak şirketin tanımlanmasıyla, yatırım önceliklerinin belirlenmesiyle ve diğer işletme ve alt birimlere kaynak dağıtımının yapılmasıyla ilgilidir. Stratejik yönetim bir örgütü hedeflerine ulaştıracak kararların fonksiyonlararası uyum içinde biçimlendirilerek uygulamaya koyulması sürecini ifade etmektedir. Bu tanımdan anlaşılacağı üzere her hangi bir işletme fonksiyonunu yerine getirebilmek için ilgili alana ait uzmanlık becerisinin sergilenmesi yeterli olabilecekken stratejik yönetim için üretimden pazarlamaya, muhasebeden insan kaynaklarına işletmenin tüm fonksiyonlarını sentezleyebilecek bir kavramsal yetenek gerekmektedir. Dolayısıyla bugünkü anlamıyla stratejik yönetim tüm işletme fonksiyonları arasındaki uyum ve ahengi sağlayan bir orkestra şefi görevini yerine getirmektedir. Dolayısıyla stratejik yönetim dar bir planlama anlayışını değil rekabetçi bir ortamda örgütün ayakta kalmasını sağlayan dinamik bir yönetim sürecini ifade etmektedir (Coşkun, 2016). Stratejik yönetimin başarısı için bütüncül ve sistematik bir yaklaşım son derece önemlidir. 1.1. Stratejik Yönetimin Seviyeleri Her bir işletme fonksiyonun genel işletme stratejisi içindeki rolü ve önemi yadsınamaz. Ancak bu çalışmada taklit ve ikame edilmesi mümkün olmayan, ulaşılması her zaman kolay olmayan, ulaşılsa bile kaybetmemek için ayrı bir çaba ve özen isteyen ve örgüte ciddi bir rekabet üstünlüğü kazandıran insan sermayesi unsurunun genel örgüt stratejisi içindeki yeri ve önemi ele alınmıştır. 2. Stratejik İnsan Kaynakları Yönetimi Stratejik yönetim anlayışının yaygınlaşması ve örgütlerde iş stratejilerinin uygulanmasıyla stratejik insan kaynakları yönetimi de stratejik yönetimin önemli bir unsuru olarak yerini almıştır. Örgütün genel stratejik eylem ve planlarında insan kaynakları yönetiminin stratejik önemi 1980’li yılların başından itibaren anılmaya başlamış bir olgudur (Akdoğan ve Cingöz, 2015). Stratejik İKY, örgütün iş hedeflerine insanlar aracılığıyla nasıl ulaşılacağına ilişkin niyetlerini ve planlarını tanımlamaktadır. Kaynak temelli yaklaşımın temel varsayımı, İKY’ nin katma değer yaratarak, örgütün nadir, taklit ve ikame edilmesi zor insan kaynaklarının stratejik geliştirilmesi ile sürdürülebilir rekabet avantajının yakalanmasına destek olmaktır (Armstrong, 2017: 18). Dolayısıyla stratejik insan kaynakları yönetimi bir örgütün insan kaynakları uygulamalarının örgütün stratejik amaçlarıyla uyumlaştırılması süreci olarak ifade edilebilir. Öte yandan stratejik İKY, örgüt kültürünün yenilik, esneklik ve çevikliği teşvik edecek şekilde dizayn edilerek örgütsel performansın iyileştirilmesi için örgütün genel stratejik amaçları ve insan kaynakları uygulamaları arasında bütünleşmeyi sağlama süreci olarak da tanımlanabilir 202 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi (Dessler, 2000). Stratejik insan kaynakları yönetiminin dayandığı temel varsayımlar şu aşağıdaki gibi ifade edilerbilir (Armstrong,2017): (Dessler, 2000). Stratejik insan kaynakları yönetiminin dayandığı temel varsayımlar şu aşağıdaki gibi ifade edilerbilir (Armstrong,2017): • Örgütün sahip olduğu insan kaynağı örgütün genel performansı üzerinde stratejik bir rol oynamaktadır • İnsan kaynakları strateji ve planları örgütün genel strateji ve planlarıyla bütünleştirilmelidir • İş stratejisini yürüten insanlardır • İK stratejilerini başarılı bir şekilde hayata geçirmek için sistematik bir bakış açısı geliştirilmelidir Öte yandan yazın incelendiğinde stratejik insan kaynakları yönetimine sonuç ve süreç odaklı ele alan iki yaklaşımın olduğu görülmektedir. Sonuç yaklaşımına göre İK yönetimine pasif bir bakış açısıyla yaklaşılmaktadır. Buna göre örgütsel strateji öncelikle belirlenir, IK süreçleri ise bu örgütsel stratejiyi benimseyip ona uyum sağlamalıdır. Süreç yaklaşımına göre ise IK fonksiyonu aktif bir rol üstlenmekte, kendi strateji ve uygulamalarıyla örgütün stratejileri arasında sinerji ve uyumu sağlamaya çalışmaktadır. Dolayısıyla sonuç ve süreç yaklaşımlarının zıt iki görüşü yansıttıkları ifade edilebilir (Akdoğan ve Cingöz, 2015) Bir örgütün rekabetçi konumunun belirlenmesinde sahip olduğu entelektüel sermayesi, üstün performans, verimlilik, esneklik ve yüksek düzeydeki sosyal uyum becerileri gibi nitelikler insan sermayesinin en önemli bileşenleri olarak tanımlanmaktadır. İnsan kaynaklarına stratejik yaklaşılmasının ve genel işletme stratejisiyle bütünleştirilmesi ihtiyacının ana gerekçelerinden biri kısa sürede değişebilen rekabet koşullarıdır. Bu bütünleşmenin oluşmasını gerekli kılan çeşitli faktörler söz konusu olmaktadır. 2. Stratejik İnsan Kaynakları Yönetimi Bunlar şöyle ifade edilebilir (Barutçugil, 2004): • Rekabetin yoğunlaşması, • Teknolojik değişimin hızının artması, • Demografik yapıda meydana gelen değişimler • Ekonomik belirsizlik ve dalgalanmaların yoğunlaşması • Örgütlerin yeniden yapılandırılmasındaki yoğunlaşmalar Bu çerçevede stratejik İKY, örgütsel performansı yükseltmek ve sürdürülebilir rekabet avantajı elde edebilmek için çeşitli İK uygulamalarının hem birbiriyle hem de örgütün stratejik kararlarıyla bütünleşmesi olarak tanımlanabilir (Wright ve McMahan, 1992). 203 Fazilet Nohut / Özlem Balaban 2.1. Stratejik İnsan Kaynakları Yönetiminde İşlevsel Taktikler Her ne kadar 1980’li yıllardan itibaren stratejik yönetimin gündemine girmeyi başarmış olsa da stratejik İnsan kaynakları yönetimi 1990’lı yıllarda önemli bir destek göremeye başlamıştır. İnsan kaynaklarındaki paradigma değişimi personel giderlerine insan merkezli bir yatırım olarak bakmayı gerektirmektedir. Yani bir diğer ifadeyle, örgütün insan sermayesini değer yaratmak için ele almasını ve işletmenin değer zincirinde önemli bir rolü olduğunu kabul etmesi anlamına gelmektedir. Stratejik insan kaynakları yönetimi şirketin insan kaynağından üst düzeyde yararlanarak hem şirketin kısa vadeli hedefleri hem de çalışan memnuniyeti ve gelişimine destek vererek, uzun vadeli başarıların elde edilmesini sağlar. Stratejik insan kaynakları yönetiminin başarılı bir şekilde uygulanabilmesi şirketler çalışan ilişkilerine yön veren bir takım taktiklere ihtiyaç duyarlar. Bu taktiklerin işlevsel olabilmesi için yanıtlaması gereken temel soruları aşağıdaki tabloda sınıflandırılmıştır (Pearce ve Robinson, 2015:292). İşlevsel Taktik İşlevsel Taktiğin Yanıtlaması Gereken Sorular İşe alım, seçme ve yerleştirme Seçilen stratejiyi desteklemede ne tür önemli insan kaynaklarına ihtiyaç vardır? Bu insan kaynakları nasıl en etkili şekilde işe alınabilir? Seçim süreçleri nasıl ileri derecede geliştirilmiş olmalıdır? Yeni çalışanlar kuruma nasıl tanıtılmalıdır? Kariyer gelişimi ve Eğitim Gelecekteki ihtiyaç duyulabilecek insan kaynağı özellikleri nelerdir? Bu ihtiyaçları karşılamak üzere nasıl bir yol izlenmelidir? Çalışanların gelişimine nasıl yardım edilebilir? Tazminat Talep edilen görevler için hangi seviyede ödemeler uygundur? İyi çalışanlar nasıl motive edilebilir ve elde tutulabilir? Maaş, teşvik, kıdem ve ekstra fayda politikaları nasıl yorumlanmalıdır? Değerlendirme, disiplin ve kontrol Çalışanlar sıklıkta değerlendirilmelidir? Değerlendirmeler resmi olarak mı gayri resmi olarak mı yapılmalıdır? Düşük performans ve uygunsuz davranışlarla ilgili olarak ne tür disipliner adımlar atılmalıdır? Bireysel performans ve grup performansı hangi yollarla kontrol edilmelidir? Çalışma ilişkileri ve fırsat eşitliği Çalışan dayanışması nasıl büyütülebilir ? Farklılıkların yönetimi konusunda istihdam politikaları oluşturulmuş mudur? Ayrım gözetmeksizin hakkaniyet ve liyakat temelli fırsat eşitliği sağlanıyor mu? Kaynak:Pearce II, John A., Robinson,Jr., B., Stratejik Yönetim, Geliştirme, Uygulama ve Kontrol, McGrawHill Irwin, Çeviri Editörü: Prof.Dr. Mehmet Barca, On ikinci Basımdan Çeviri, Nobel Yayın, s.29 204 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi 2.2. Stratejik Uyum Stratejik İKY, dikey olarak iş stratejisiyle ve yatay olarak birbiriyle entegre edilmiş İK stratejilerinin geliştirilmesine yönelik kapsayıcı yaklaşımların kullanılmasını içeren bir süreçtir. Bu stratejiler, örgütsel etkinlik gibi genel kurumsal hususlarla ve kaynak sağlama, öğrenme ve geliştirme, ödül ve çalışan ilişkileri gibi insan yönetiminin daha spesifik yönleriyle ilgili niyetleri ve planları tanımlamaktadır (Armstrong, 2006) Dolayısıyla stratejik insan kaynakları yönetimi yatay ve dikey boyutları olmak üzere çift boyutlu bir yapıyı ifade etmektedir. Bazı kaynaklarda iç uyum olarak da adlandırılan yatay uyum insan kaynakları yönetimi faaliyetlerini bir sistem olarak ele alan ve bu sistemi oluşturan her bir unsurun birbiriyle senkronize bir şekilde çalışmasını ifade eden bir yapıdır. Bir diğer ifadeyle seçme yerleştirme, eğitim, kariyer planlama, performans değerlendirme ve ücretlendirme gibi İnsan kaynakları yönetimi süreçlerini oluşturan her bir adımın birbiriyle uyumlu ve tutarlı bir biçiminde yönetilmesi sonucunda ortaya çıkacak İK performansının etkisinden söz edilmektedir. Bir sistem olarak ele alınan sürecin başarılı bir şekilde gerçekleşmesi yatay uyumu ifade etmektedir ve bu uyum sağlandığı takdirde İKY’ nin işletme performansına olumlu etkilerde bulunabileceği varsayılmaktadır (Akdoğan ve Cingöz, 2015; Balaban ve Palaz, 2018). Dikey uyum ya da bir diğer ifadeyle dış uyum kavramı ise İnsan kaynakları yönetim süreçlerinin bir bütün olarak örgütün diğer fonksiyonlarıyla uyum içinde çalışmasını ifade etmektedir. Dikey uyumda örgüt başarısına etki edecek faktör fonksiyonel uyumdur. Yani IK uygulamalarının örgütün stratejik hedef ve stratejik gereksinimleriyle bütünleşecek şekilde yapılandırılması gerekmektedir. Gerek dikey uyum gerekse yatay uyumun sağlanabilmesi için öncelikle IK yöneticilerinin örgütün genel amaç ve hedeflerine ulaşması konusunda IK süreçlerinin ne gibi bir rol ve öneme sahip olduğunu anlamaları gerekmektedir. Bu anlayışa sahip olmakla birlikte fırsat ve tehditleri görebilme, esnek hareket edebilme kabiliyeti, mevcut kaynakları doğru yönetme ve olası ihtiyaçları tahmin edebilme yetkinliğine de sahip olmaları gerekmektedir. Dolayısıyla gerek yatay uyumun, gerek dikey uyumun sağlanması ve etkin fonksiyonel stratejilerin oluşturulabilmesi için bir IK yöneticisinin stratejik rollerini aşağıdaki gibi ifade etmek mümkündür: • IK stratejini belirleyecek üst kademe IK yöneticileri IK stratejilerinin oluşturulması sırasında mevkidaşlarıyla (diğer hat yöneticileriyle) sıkı bir iletişim ve iş birliği içinde olmalıdırlar. 205 Fazilet Nohut / Özlem Balaban • Üst kademe IK yöneticileri diğer üst kademe yöneticilerin, CEO, CFO, CMO gibi üst düzey yöneticilerin katıldığı tüm toplantılara katılmalıdır. Böylelikle örgütün gündeminde olan stratejik konulara insan kaynakları perspektifinden değerlendirme fırsatı bulur. • IK yöneticileri fonksiyonel stratejilerini amacına uygun bir şekilde uygulayabilmek için diğer hat yöneticileriyle iletişim ve işbirliği içinde olmalıdır. Böylelikle dikey uyumun etkin bir şekilde gerçekleşmesine olanak bulunur. 2.2. Stratejik Uyum • Fonksiyonel stratejilerin de etkin bir biçimde uygulanabilmesi için IK bölümü kendi iç süreçlerini stratejik olarak yönetebilmelidir. Böylelikle de yatay uyum sağlanmış olacaktır (Armstrong, 2017). Stratejist bir üst düzey IK yöneticisinin yukarıda bahsedilen rollerine ilave olarak örgütün genel performansına katkı sunacak doğru stratejileri bütüncül bir bakış açısıyla belirleyebilmesi için gereksinim duyduğu bilgileri aşağıdaki yollarla elde edebilir: • Müşteriler, ürün ve hizmetleri deneyimleyen kullanıcılarla iletişim kurarak ihtiyaçları tespit etmek • Dünya standartlarında yönetim uygulamalarının sağlanması konusunda ve değişim yönetiminde iş bilgisi kabiliyetini göstermek • Örgütün faaliyet gösterdiği bağlamı anlamak ve gereksinimlerinin farkında olmak • Teknolojik, finansal, ekonomik, sosyal gelişmeleri yakından takip etmek ve iş toplantılarına aktif bir biçimde katılarak örgütün sahip olduğu yetenekler hakkında bilinçli katkılar sunmak Tüm bu bilgilerin yanı sıra IK stratejilerinde başarılı olabilmek için bütünleşme ve uyum sürecinde önemli rol oynayan kavramsal değişkenlerin bilinmesinde de fayda vardır. Bu kavramsal değişkenler aşağıdaki başlıkta açıklanmıştır. 2.3 Stratejik İnsan Kaynakları Uyum Sürecinin Kavramsal Değişkenleri Stratejik İKY’nin örgüt stratejisiyle İKY uygulamalarını uyumlaştıran bir kapsamda temellendiği ifade edilmektedir. Bu anlamda, strateji-İKY uyum üzerine yapılan çalışmalarda farklı bakış açılarıyla benzer açıklamalar yapılmıştır. Wright ve Snell (1998), stratejinin üç genel kavramsal değişkene uyması gerektiğini ileri sürmektedir. Bunlar sırasıyla “İKY uygulamaları”, “çalışan becerileri” ve “çalışan davranışları” dır (Wright ve Snell, 1998: 758). 206 \| Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi Strateji-İnsan Kaynakları Uyumu: Schuler ve Jackson (1987), örgütlerin ellerinde bir İKY uygulamaları “menüsüne” sahip olduğunu öne sürmektedir; farklı örgüt stratejileri, çalışanlardan farklı rol davranışları gerektirmektedir. Bu nedenle örgütler, seçilen bir stratejiyi uygulamak için gerekli davranışları ortaya çıkarma yeteneklerine dayalı olarak İKY uygulamalarını seçmektedirler. Benzer şekilde, Miles ve Snow (1984) savunmacı, araştırmacı ve analizci stratejik tipler arasında gözlemlenebilecek İKY uygulamalarındaki farklılıkları tartışmaktadır. Wright ve Snell (1991), hem firmanın stratejik tipinin hem de yönlü stratejisinin (geri çekilme, büyüme vb.) İKY uygulamaları setinin seçimini etkileyeceğini belirtmektedir (Patrick ve Snell 1998: 758). Strateji-Çalışan Becerileri Uyumu: İşletme stratejisi ile çeşitli çalışanların becerileri veya özellikleri arasında uyumu ifade edilmektedir. Örneğin, yönetim kurulu başkanı ve genel müdürler arasındaki eşleşme ile ilgili birçok reçete önerilmektedir. Bu anlayış “farklı stratejiler, etkili performans için farklı tipte insanlar gerektirir” varsayımına dayanmaktadır (Patrick ve Snell,1998:758). Strateji-Çalışan Davranışı Uyumu: Örgütün stratejisi ile çalışanlar tarafından sergilenen davranış türleri arasında bir uyum olduğu ifade edilmektedir. Davranışsal bakış açısına göre farklı stratejiler farklı rol davranışları gerektirmektedir (Patrick ve Snell, 1998: 758). Stratejik İKY yazını genel örgüt stratejisi ve İKY uyumu ve İKY ve firma performansı arasındaki etkileşimle ilgilenmektedir (Panayotopoulou ve Papalexandris, 2003: 681). Stratejik İKY iki stratejik uyumun gözetilmesi gerektiğini ileri sürmektedir. Birincisi dış uyum yani, birimlerin yapısı, sistemleri ve tek elden yönetim uygulamaları örgütün gelişim stratejisine uygun olmasını ifade etmektedir. İkincisi ise iç uyum yani ik iç süreçlerinin birbiriyle uyumu, kendi iç politika ve kararlarının genel stratejiyi destekleyecek nitelikte olmasını ifade etmektedir (Baird ve Meshoulam, 1988:116). Uyum ve bütünleşme sürecinde önemli rol oynayan kavramsal değişkenlere değindikten sonra dışsal uyum ve içsel uyum süreçlerinin yöntem ve aşamalarını detaylı bir şekilde açıklamakta fayda vardır. Aşağıdaki başlıkta sırasıyla uyum süreçleri açıklanmıştır. Yaşam Döngüsü ve Hiyerarşik Modeller Gelişimin, doğum, olgunluk ve düşüşün tahmin edilebilir, nispeten farklı “yapı bloklarında” meydana geldiği yaşam döngüsü modelleri ve katı bir yaşam döngüsü yaklaşımını benimsemese de örgütsel büyüme ve gelişmenin, önceki aşamaların sonraki aşamalar için temel oluşturduğu yapı taşlarında meydana geldiğini savunulmaktadır. Evrimsel Modeller Evrimsel modellerde örgütlerin ölümü içeren bir yaşam döngüsü izlemediklerini, ancak çevreye uyum sağlamak için kendilerini reforme ettiklerini ve yenilediklerini savunmaktadır. Bunu yapmayan örgütler gerilemekte ve ölmektedir. Bu modellerde, örgütün çevreye tepki vermesine ve çevreye uyum sağlamasına yardımcı olan evrimsel güçlere odaklanılmakta, yaşlanmaya odaklanılmamaktadır. 2.4. Dışsal Uyum Yapının, sistemlerin ve yönetim uygulamalarının bir örgütün gelişim aşamasına uygun hale getirilmesi önemlidir. Örgüt büyüdükçe ve geliştikçe değişime ihtiyaç duymaktadır. Bir örgütün büyüdükçe nasıl değiştiğini anlayarak, insan kaynakları yönetimi uygulamalarının nasıl değişmesi 207 Fazilet Nohut / Özlem Balaban gerektiğini anlaşılabilmektedir. Örgütün büyümesini ve gelişimini açıklamak için yaşam döngüsü/hiyerarşik, evrimsel, aşama ve metamorfoz olmak üzere dört tür model kullanılmaktadır: Bu modellerin hepsi örgütsel ihtiyaçların nasıl değiştiğini ve insan kaynakları yönetiminin bunlara yanıt vermek için ne yapması gerektiğini anlamak için yararlı olmaktadır. Bu modeller sırasıyla aşağıda açıklanmıştır (Baird ve Meshoulam, 1988). Aşama Modelleri Aşama modellerinde örgütlerin çevrelerine uyum sağlamasına rağmen, sürecin tamamen tepkisel olmadığı ifade edilmektedir. Sıralı yapı taşları ve çevresel yanıt verme, aşama modellerine dahil edilmekte ve buna, yönetimsel eylemin örgütün çevreye uyum sağlamasına izin verdiği fikri eklenmektedir. Yöneticilerin strateji ve yapıyı çevreye uyacak şekilde uyarlamaları önerilmektedir Metamorfoz Teorisi Aşama modelleri “Örgüt bir aşamadan diğerine nasıl geçer?” sorusunu yanıtsız bırakmaktadır: Metamorfoz modelleri, değişimin örgüt ile çevre arasındaki uyumun örgütün etkinliğini ve hayatta kalmasını tehdit edecek kadar kötü olduğunda meydana geldiğini öne sürmektedir. Değişim, küçük artımlı adımlar yerine büyük değişimlerle meydana gelmektedir. Bu teoriler, örgütsel büyüme ve gelişmenin temel özelliklerini tanımlamaktadır. İnsan kaynakları yönetiminin etkili olması ve örgüte uyması için, büyümesinin bir parçası olduğu örgütün büyümesine paralel olması gerekmektedir ve aşağıdaki hususlara dikkat edilmelidir: 208 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi • Baskılara ve fırsatlara yanıt olarak öngörülebilir bir sırayla değişmeli ve gelişmelidir. • Aşamalardaki değişim, her aşamada insan kaynakları yönetimine kesinlikle farklı bir yönelim sergilemektedir. • Her aşama önceki aşama üzerine inşa edilmeli ve bütünleştirilmelidir. • İnsan kaynakları yönetimini örgütün ihtiyaçlarına uydurmak için proaktif olarak çalışılmalıdır. İnsan kaynakları yönetiminin etkinliği, örgütün gelişim aşamasına uygunluğuna bağlıdır. Örgüt büyüdükçe ve geliştikçe, insan kaynakları yönetimi programları, uygulamaları ve prosedürleri ihtiyaçlarını karşılamak için değişmeli ve geliştirilmelidir. Büyüme ve gelişme modelleriyle tutarlı olarak, örgüt daha karmaşık hale geldikçe insan kaynakları yönetiminin bir dizi aşamadan geçtiği söylenebilir. Her aşama, önceki aşamaları birleştirmekte ve üzerine inşa etmektedir. 2.5. İçsel Uyum İnsan kaynakları yönetimi sadece örgütün gelişim aşamasına uymamalı, aynı zamanda insan kaynakları yönetiminin bileşenleri de birbiriyle uyumlu olmalı ve birbirini desteklemelidir. Yani bir diğer ifadeyle ik süreçlerinin yapılandırılması, görev ve sorumlulukların uyumu, insan ilişkileri ve idari süreçler arasındaki uyum örgütsel performansı üzerinde olumlu bir rol oynamaktadır. Dolayısıyla insan kaynakları uygulamaları hem kendi içerisinde hem de örgütün gelişim amaçlarıyla uyum içerisinde olmalıdır. İç uyum, insan kaynakları yönetiminin altı stratejik bileşeni üzerinde durmaktadır. Bu altı stratejik bileşen aşağıda sırasıyla açıklanmıştır (Baird ve Meshoulam, 1988: 122-123). • Yönetim Bilinci: Yönetim farkındalığı, işe alma ve işten çıkarma gibi idari ihtiyaçlara odaklanmaktan, tüm yönetim karar alma süreçlerinde insan kaynakları hususlarının tam entegrasyonuna kadar uzanmaktadır • Fonksiyonel Yönetim: Bu bileşen, insan kaynakları fonksiyonunun yapısını ve kaynaklarının planlanmasını, tahsisini ve kontrolünü içermektedir. Yapı, çok gevşek veya merkezi olmayana kadar değişebilmektedir • Program Portföyü: Program portföyü, maaş yönetimi ve kayıt tutma programlarından çok karmaşık ve sofistike esnek ücretlendirme, çevresel tarama ve uzun vadeli planlama programlarına kadar uzanmaktadır 209 Fazilet Nohut / Özlem Balaban \| • Personel Becerileri: Personel profesyonellerinin uygun becerilere ihtiyacı vardır. Temel programlar ve basit bilgi sistemleri temel beceriler gerektirir. Karmaşık programların eklenmesi ve boyut olarak büyüme, daha gelişmiş, farklılaştırılmış ve uzmanlaşmış beceriler gerektirmektedir • Personel Becerileri: Personel profesyonellerinin uygun becerilere ihtiyacı vardır. Temel programlar ve basit bilgi sistemleri temel beceriler gerektirir. Karmaşık programların eklenmesi ve boyut olarak büyüme, daha gelişmiş, farklılaştırılmış ve uzmanlaşmış beceriler gerektirmektedir • Bilgi Teknolojisi: Bilgi araçları, manüel kayıt tutmadan, modelleme yetenekleri olan gelişmiş dağıtılmış sistemlere kadar çeşitlilik göstermektedir. Bilgi teknolojileri, resmi analitik araçlardan istatistiksel araçlara dayalı gelişmiş tahminlere ve simülasyona kadar uzanmaktadır • Çevre Farkındalığı: İlk aşamada, örgüte olan baskılar nedeniyle yönetim, çevreyi sistematik olarak değerlendirmemekte ve tepki vermemektedir. Bu aşamada yönetim, iç ve dış çevrenin ve bunların etkilerinin farkında olmaktadır. Esnek kalmakta ve ortaya çıkan fırsat ve risklere göre uyum sağlamaktadır İnsan kaynakları yönetimi birimi, altı stratejik bileşenin tümü aynı gelişim aşamasında olduğunda en iyi şekilde çalışmaktadır (Baird ve Meshoulam, 1988). Bu bileşenler birbiriyle eşleşmez ise para, zaman ve enerji gibi kaynaklar boşa gitmiş olmaktadır. Örneğin personel becerilerinin fazla gelişmesi, mevcut kişi-iş uyumunu azaltarak çalışanlarda tatminsizliğe ve daha yüksek maaş beklentisine yol açabilir. Bilgi teknolojilerine yapılan fazla yatırım diğer bileşenlerle eşleşmediği zaman kaynak israfına yol açabilir. Bahsedilen herhangi bir bileşenin fazla geliştirilmesi veya az geliştirilmesi sistemsel soruna yol açacaktır. 2.6. Stratejik İnsan Kaynakları Yönetiminin Önemi Başarılı bir stratejik yönetim örgütün her düzeyinden yöneticilerin iletişim ve işbirliği içinde olarak planlama ve uygulama becerilerindeki uyumu ifade etmektedir. Bir diğer ifadeyle stratejik yönetim uygulamalarının katılımcı karar alma mekanizmasının sonucunda ortaya çıkan davranışsal sonuçlara benzer katkıları vardır. Dolayısıyla stratejik yönetim uygulamalarının örgütsel performansa etkisinde sadece mali unsurların değil, mali olmayan davranış temelli etki ölçülerinin de dikkate alınması gerekmektedir. Bu davranış temelli faydalardan bazılarını aşağıdaki gibi ifade etmek mümkündür: - Stratejik planlamanın önemi hakkında astlarını bilinçlendiren, destekleyen ve güçlendiren yöneticiler astlarının izleme ve tahmin yeteneklerinin gelişmesini sağlarlar. Astların bu yeteneği stratejik kararların alınmasında önemli veri kaynağı olarak değerlendirilebilir. Böylelikle örgütün sorun önleme yeteneği gelişir. \| Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi 210 - Katılımcı karar alma mekanizması sayesinde ortaya çıkan grup etkileşimi alternatif stratejiler üretebilir. Dolayısıyla grup üyelerinin uzmanlaşmış bakış açılarına dayanan öngörüler farklı alternatiflerin değerlendirilmesini ve bu alternatifler arasından örgüt için en iyi stratejik kararın alınmasını kolaylaştıracaktır. - Katılımcı karar alma mekanizması sayesinde ortaya çıkan grup etkileşimi alternatif stratejiler üretebilir. Dolayısıyla grup üyelerinin uzmanlaşmış bakış açılarına dayanan öngörüler farklı alternatiflerin değerlendirilmesini ve bu alternatifler arasından örgüt için en iyi stratejik kararın alınmasını kolaylaştıracaktır. - Öneri ve görüşlerinin örgütün stratejik kararlarında önemli bir katkı olarak değerlendirildiği bilen çalışanlar üretkenlik-ödül ilişkisine dair bir anlayış geliştirebilir ve bu yolla da yüksek motivasyona sahip olabilirler. - Strateji oluşturma süreçleri rol tanımlamalarını şeffaf bir şekilde ortaya konulmasını gerektirdiğinden çalışanların görev, yetki ve sorumlulukları açısından belirsizlik yaşanması önlenecek ve rol çatışmaları azalacaktır. - Katılımcı bir stratejik yönetim değişime direnci önemli ölçüde azaltacaktır. Çünkü kararlara katılan bireyler, her ne kadar sonuçlardan memnun olmasalar bile nihai kararları etkileyen, kısıtlayan parametrelerin farkında oldukları için otoritenin kararına nispeten katılım gösterdikleri kararları kabul konusunda daha uyumlu davranacaklardır. Katılımcı stratejik yönetim sürecinde göz önünde bulundurulması gereken bir takım unsurlar vardır. Bunlardan birisi yöneticilerin operasyonel görevlerini yerine getirmek için gereken zaman ve stratejik kararlara alma konusundaki harcadıkları zaman arasındaki dengeyi kurabilmeleredir. Bir diğer önemli husus ise katılımcı astların gerçekleşmeyen beklentileri karşısında yaşayabilecekleri olası hayal kırıklıklarına karşı yöneticilerin hazırlıklı olması ve bu süreci doğru yönetebilecek beşeri yetkinliklere sahip olmalarıdır. Nitekim stratejik karar alma sürecinin en küçük aşamasına bile katılmış olan bir çalışan çeşitli beklentiler içine girebilir. Örneğin birey önerisinin kabul edilmesi, katkılarının ödüllendirilmesi gibi isteyebilir gerçekleşmediğinde ise hayal kırıklığı ya da küskünlük yaşayabilir. Dolayısıyla bu sürecin çalışma barışını bozmayacak şekilde yönetilmesi stratejik planlamanın potansiyelini önemli ölçüde etkileyecektir (Pearce ve Robinson, 2015:9). 3. Genel İşletme Stratejileri ve Stratejik İnsan Kaynakları Yönetiminin Bütünleştirmesi Strateji, örgütlerde değişimi ve gelişimi sağlayan, sürekli olarak örgütün çevre ile uyum içinde olmasını kolaylaştıran, beklenmedik durumları kontrol altına alan bir yönetim mekanizmasıdır. Stratejik yönetimde, örgütler faaliyetleri ile çevredeki değişimleri izleyerek örgütün güçlü ve zayıf 211 Fazilet Nohut / Özlem Balaban yönlerine sahip olduğu kaynakları değerlendirerek karar verilmesi gerekir. Bu durumun neticesinde alınan karalar örgütün bilgi temelli dinamik ve gelişmelere açık bir yönetim anlayışı içinde çalışmasını sağlayacaktır. Böylece gelecekte var olabilecek değişimlerin sistematik bir şekilde öngörülmesi daha rahat olacaktır. Sonuç olarak örgütün büyümesi, başarılı olması gelişigüzel olmayacak, planlanmış ve kontrollü bir şekilde gerçekleşecektir (Barutçugil, 2004: 54-55). Dolayısıyla örgütler çevrelerini izleyerek gerekli değişimleri yapmak ve başarılarının sürdürülebilir olması için rekabet stratejileri geliştirmişlerdir. Bu rekabet stratejilerinin ve stratejik IK yönetiminin uyumuna aşağıdaki başlıklarda değinilmiştir. 3.1 Rekabet Stratejileri Örgütler hem çevrelerinden etkilenen hem de çevrelerini etkileyen, organik bir yapıya sahip sistemlerdir. Örgütlerin varlığı öncelikle rakipleri tarafından etkilenirken, sosyal, politik, ekonomik değişkenler, müşteriler, tedarikçiler ve yasalar tarafından da etkilenmektedir. Dolayısıyla böylesine değişken, dinamik ve belirsizliğin hakim olduğu bir çevreye uyum sağlayabilmek ve ayakta kalabilmek için işletmelerin rekabet stratejileri geliştirmeleri gerekmektedir. Rekabet, aynı veya benzer sektörde faaliyet gösteren örgütlerin birbirlerine olan üstünlüğü olarak ifade edilmektedir. Rekabet, mevcut ve potansiyel müşterilerin ihtiyaç ve beklentilerini karşılamak amacıyla, katma değerli ürün ve hizmetler sunarak ilgili pazarlarda etkin bir mücadele olarak da tanımlanabilir (Ülgen ve Mirze, 2004). Ancak rekabet üstünlüğü elde etmek kadar bunun sürdürülebilir olması da son derece önemlidir. Nitekim rekabet üstünlüğünün sürdürülebilir olmasını her örgüt sağlayamamaktadır. Bu sebeple Porter (1985), bazı örgütlerin diğerlerine göre daha uzun süre başarılı olmalarını sağlayacak, yazında jenerik stratejiler olarak da anılan toplam maliyet liderliği, farklılaşma ve odaklanma stratejilerini önermektedir. Örgütlerin benimsediği jenerik stratejilerin gerektirdiği belli davranış ve rol biçimleri bulunmaktadır. Bunlar hem ilgili rekabet stratejisini uygulayan genel işletme yapısını hem de o yapının istihdam politikasındaki stratejik unsurlara ilişkin yapıyı açıklamaktadır (Porter, 1985). Aşağıdaki alt başlıklarda bu rekabet stratejileri ve işletme-iky uyumuna ilişkin açıklamalara yer verilmiştir. 3.1.1 Farklılaştırma Stratejisi ve İnsan Kaynakları Yönetimi Bir örgüt, alıcılar tarafından değer verilen çeşitli boyutlarda benzersiz olarak algılanan bir ürün/hizmet sunduğunda farklılaştırma stratejisi izlediği düşünülmektedir (Porter, 1985). Farklılaşmanın ana kaynağı ürün/hizmet yeniliğidir. İnovasyon yoluyla farklılaşma peşinde koşan örgütlerin yenilikçi 212 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi olmaları ve çevrelerine uyum sağlamaları gerekmektedir. Örgütler nitelikli iş görenleri seçtiğinde ve onlara özerklik verdiğinde, performanslarını uzun vadeli hedefler temelinde değerlendirdiğinde onlara deneyerek hata yaparak öğrenme fırsatı sunmaktadır. Öte yandan inovasyonun bürokrasi tarafından sürekli kısıtlandırıldığı zaman ise açık iletişim ve sürekli öğrenme ile iş görenler güçlendirilmektedir (Aktaran Panayotopoulou ve Papalexandris, 2003: 686). İK fonksiyonunun inovatif örgütlerde, yüksek düzeyde esneklik gerektiren ve açık sistem anlayışı kapsamında değişime açık bir yapıda hareket etmesi gerektiği önerilmektedir. Başka bir farklılaştırma stratejisi türünde ise, ürünlerin ve süreçlerin sürekli iyileştirilmesi amaçlanmaktadır. Bu durumda, örgütün temel ilgi alanı entelektüel sermayedir ve çalışanların bilgi işçisi olmaları ve planlama, kalite kontrol, problem tanımlama ve problem çözme süreçlerine katılmaları gerekmektedir. Kalite stratejisinin özü, insan ilişkileri modeline uyan bir özellik olan yetkinlik edinme ve geliştirmeye dayanmaktadır. Farklılaşma stratejisi; esneklik, yenilikçilik, kalite, müşteri hizmetleri ve proaktifliğe dayalı olduğundan, daha iyi örgütsel büyüme ve pazar performansı göstermeleri beklenir. Burada esneklik ile örgütsel ve piyasa performansı arasında pozitif bir ilişki olduğu ifade edilmektedir (Panayotopoulou ve Papalexandris, 2003:687). 3.1.2 Maliyet Liderliği Stratejisi ve İnsan Kaynakları Yönetimi Porter (1980)’a göre, maliyet liderliği stratejisi izleyen firmalar, pazardaki en düşük maliyetle ürün ve hizmet üretip, sunmayı amaçlamaktadır. Porter’ın maliyet odaklı stratejisi, tekrarlayan ve tahmin edilebilir davranışları, kısa vadeli ve çıktı odaklı olmayı, temel kaygının sonuçlar olduğu, risk seviyesi düşük faaliyetleri, istikrarı tanımlamaktadır. Maliyet liderliğine uygun diğer özellikler, sınırlı ve uzmanlaşmış görevlere dayalı iş organizasyonu, tekrarın ve rutin görevlerin artması, prosedürlere sıkı bağlılık ve sıkı kontrolü içermektedir. Açıkça tanımlanmış performans standartlarının varlığı İK işlevini, rasyonel hedef modelinin bir özelliği olan çıktı kontrolünü vurgulamaya zorlamaktadır. Dolayısıyla kurallar, politikalar, hiyerarşi ve diğer bürokratik kontrol mekanizmalarının kullanımının maliyet liderliği stratejisi ile ilgili olduğunu belirtilmektedir. Maliyet liderliği stratejisi izleyen firma, genel giderlerin en aza indirilmesi ve ölçek ekonomileri ile karakterize edilmektedir. Bu nedenle olumlu finansal sonuçlar göstermeleri beklenmektedir (Panayotopoulou ve Papalexandris, 2003: 687). 213 Fazilet Nohut / Özlem Balaban 3.1.3 Odaklanma Stratejisi ve İnsan Kaynakları Yönetimi Porter (1980), odaklanma stratejisiyle, örgütün dikkatini belirli bir müşteri, ürün veya coğrafi bölge üzerinde hizmet etmeye odaklayan bir niş stratejisi belirlemeyi ifade etmektedir. Örgüt, endüstrinin uzmanlaşmış bir bölümünde ya bir farklılaştırma ya da bir maliyet liderliği stratejisi (veya ikisinin bir kombinasyonu) kullanmaktadır. Maliyete dayalı bir odaklanma stratejisi izleyen örgütler “uzman” olarak tanımlanmakta ve sınırlı hedef grupları nedeniyle cazip ürünler/hizmetler üretirken; üretim, pazarlama ve dağıtımda ekonomi elde ettiklerini savunmaktadırlar. Bu şekilde, rasyonel hedef modeline uyan tüm niteliklerin olan kaynak etkinliği, üretkenliği ve performansı vurgulanmaktadır (Gomez-Meija vd., 2001; Panayotopoulou ve Papalexandris, 2003: 687). İnsan kaynakları yönetiminin örgütün performansına olumlu etkisinin en önemli varsayımı İK uygulamalarının bütünleşme, bağlılık, motivasyon ve yetkinlik gibi çalışan özelliklerini doğrudan etkiliyor olması ve bu etkinin sonuçlarının ise çalışanların örgütün finansal performansını arttıracak faaliyetlerde yüksek standart ve verimlilik motivasyonuyla hareket etmeye gönüllü olacak olmasıdır. Böylelikle örgütte bir IK değer zinciri oluşmuş olacaktır. Yani buna göre IK uygulamalarının örgütün performansı üzerinde doğrudan bir etki oluşturmadığı IK uygulamalarından etkilenen çalışanların olumlu davranışlar geliştirerek IK çıktılarını etkilediği ifade edilebilir. Bir diğer ifadeyle IK uygulamalarının örgütün finansal performansı üzerinde doğrudan bir etki oluşturmadığı ancak göz ardı edilemeyecek kadar önemli olan dolaylı bir etkiye sahip olduğu ifade edilebilir (Armstrong, 2017 :18) Yukarıda açıklanmaya çalışılan rekabet stratejileri ve IK yönetimi arasındaki ilişkide yatay ve dikey uyumun gerçekleştirilebilmesi için örgüt yapısına göre İK yönetiminin şekillenmesi gerektiği görülmektedir. Ancak bu yolla stratejik uyum ve olumlu örgütsel performansın ortaya çıkabileceği ifade edilebilir. Yukarıdaki açıklamalara bakıldığında rekabet stratejisine göre IK yönetiminde başarılı bir yatay ve dikey uyum için mekanik ve organik örgüt yapılarının önemi kendini göstermektedir. IK politikaları ve stratejilerinin doğru belirlenebilmesi için mekanik ve organik örgüt yapılarının gerektirdiği IK yapılanmasına değinmekte fayda görülmüştür. Aşağıdaki başlıkta stratejik tasarımın etkili bir şekilde gerçekleşmesinde rol oynayan örgüt yapılarına göre olması gereken IK yapılanması açıklanmıştır. 3.2. Örgütsel Tasarım ve Stratejik Yönetim İlişkisi 3.2. Örgütsel Tasarım ve Stratejik Yönetim İlişkisi Stratejik karar süreci örgütün amaç ve hedeflerine ulaşabilmek için elindeki sınırlı imkan ve kaynakların en verimli kombinasyonuyla örgütün 214 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi iç çevresiyle dış çevresi arasındaki uyumu sağlayarak sürdürülebilir rekabet avantajı elde etme süreci olarak ifade edilebilir. Bu noktada örgütün içinde faaliyet gösterdiği çevreye göre örgüt yapısının şekillenmesi konusu gündeme gelmektedir. Örgüt yapısı ve çevre ilişkisinin önemi ilk olarak 1950’li yıllarda İngiltere’de yapılan çalışmalarda ortaya konmuştur. Bu çalışmalarda örgütün içinde bulunduğu çevresel koşullar, teknoloji ve pazardaki yenilik hızı ve gerçekleştirilen işin doğasına göre örgüt yapısının şekillenme süreci incelenmeye başlanmıştır. Bu bağlamda yapılan çalışmaların öncü isimleri Burns ve Stalker (1961) özellikle yönetim sistemlerinin değişen çevresel özelliklere nasıl uyum sağladıklarını anlamaya çalışmışlardır. Araştırmaları sonucunda durağanlık ve değişkenlik özelliklerinin örgüt yapılarının önemli belirleyicileri olduğunu ortaya koymuşlardır (Keskin vd., 2016:227). Çevrenin durağanlığı veya belirsizlik ve dinamizmi örgüt yapısı üzerinde belirleyici bir rol oynamaktadır. Bu noktada mekanik örgüt yapısı ve organik örgüt yapısı olarak iki yapı ortaya çıkmaktadır. Bu iki yapının işleyişi, rekabet avantajı elde etme konusundaki belirleyecekleri stratejik kararları ve IK yapılanması birbirinden farklılık gösterecektir. Dolayısıyla bu yapıların anlaşılmasında fayda vardır. 3.2.1. Mekanik Örgüt Yapısı ve İnsan Kaynakları Yapılanması Mekanik örgüt yapısı belirsizliğin düşük durağanlığın yüksek olduğu çevresel koşullarda ortaya çıkan bir yapıdır. Durağan çevresel koşullar, belirgin kurallar, hiyerarşik bir denetim ve yüksek formelleşme düzeyine sahip bir örgüt yapısı gerektirir. Bu durumda örgütle dikey kontrol, uzmanlaşma, standart yöntemler, merkezi karar verme verimlilik odaklı bir yapı benimsemektedirler. Mekanik örgütler birer makine gibi tasavvur edilerek yüksek performans odaklı sistemlere dönüştürülebilen uzmanlaşmış bölümlerden oluşmaktadırlar. Keskin vd.(2016)’nın Tosi (2009)’un çalışmasına dayanarak tanımladıkları mekanik bir örgüt yapısının özellikleri aşağıdaki gibidir: • Uzmanlaşma : Fonksiyonel görevler uzmanlaşma esasına dayanarak farklılaşmıştır. Aynı amaç ile faaliyet gösteren diğer görevlerle aynı işlevlere sahip değildir. • Uzmanlaşma : Fonksiyonel görevler uzmanlaşma esasına dayanarak farklılaşmıştır. Aynı amaç ile faaliyet gösteren diğer görevlerle aynı işlevlere sahip değildir. • Standartlaşma: Her bir görev, işin tamamlanması için kendine özgü, sabit yöntem ve amaçlar içermektedir. • Basit entegrasyon mekanizmaları: Görev ve sorumluluklar hak, zorunluluk ve yöntemleri içerecek şekilde tanımlanmışlardır. 215 Fazilet Nohut / Özlem Balaban • Yüksek formalleşme düzeyi: Her bir görevin işlevsel rolü, zorunlulukları teknik yöntemleri resmi ve yazılı bir biçimde belirlenmiştir. • Yakın denetim: Hiyerarşik bir denetim mekanizmasına bağlı olarak her bir görev bir üst yönetici tarafından denetlenmektedir. Kontrol, yetki ve iletişim hiyerarşik bir düzene bağlıdır. • Teknik bilgiye dayalı görev: Bilginin örgüt içindeki konumu hiyerarşik yapının belirleyicisidir. • Dikey iletişim: Ast-üst arasındaki talimatların yöneticiler tarafından verildiği, dikey bir etkileşim mevcuttur. • Merkezi karar mekanizmaları: Örgütsel faaliyetler yöneticilerin belirlediği kural ve kararlar tarafından yönetilir. • Pozisyona dayalı otorite/prestij: Örgüt içerisindeki bilgi, deneyim ve beceriler genel bilgi, deneyim ve becerilerden daha önemlidir. Bu özellikler stratejik yönetim kapsamında değerlendirildiğinde özellikle belirli bir kıdem ve düzeyde olmayan yöneticilerin ve çalışanların stratejik kararlara katılım konusunda daha edilgen kalabilicekleri, sadece yukarıdan gelen emir ve talimatları uygulayarak stratejik yönetim sürecine katkıda bulunabilecekleri ifade edilebilir. Günümüz dünyasının gerçekleri göz önünde bulundurulduğunda böyle bir mekanik yapılanma içerisinde stratejik kararların genel örgütsel performans üzerindeki olumlu etkisi sorgulanmaya değer bir konudur. 3.2.2 Organik Örgüt Yapısı ve İnsan Kaynakları Yapılanması Organik örgüt yapısı değişkenliğin ve belirsizliğin yüksek olduğu çevrelerde faaliyet gösteren örgütlerin esnek, çevik, uyumlanma becerilerinin yüksek olduğu bir örgütsel yapılanmayı ifade etmektedir. Bu örgütlerde katı kural ve düzenlemeler olmadığı için katı bir denetim sistemi de bulunmamaktadır. Çalışanlar işleri konusunda izleyecekleri yolu seçerken inisiyatif hakkına ve yetkinliğine sahiptirler. Bu örgütsel yapılanma içinde keskin bir hiyerarşik yapılanma da mevcut değildir. Kararlar merkezi bir özelliğe sahip olmadığı için tüm işletme çalışanları düzeyinde kararlara katılım hakkı vardır. Organik yapıya sahip örgütler çevresel değişkenlere uyum konusunda hassas oldukları için daha az uzmanlaşma ve düşük formelleşme derecesine sahiptirler. Yatay iletişim ve koordinasyon önemlidir. Dolayısıyla çalışma grupları ve departmanlar arası işbirliği son derece önemlidir. Bu bilgilere ek olarak organik işletme yapısının özellikleri aşağıdaki gibi sıralanabilir( Keskin vd., 2016): 216 \| Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi • Bireysel Bilgi: Belirli bir görevin gerçekleştirilmesinde bireysel bilgi ve tecrübe önemlidir. • Bireysel Bilgi: Belirli bir görevin gerçekleştirilmesinde bireysel bilgi ve tecrübe önemlidir. • Esneklik: Bireysel görevler diğerleriyle etkileşimleri sonucunda sürekli olarak yeniden tanımlanır ve güncellenirler. • İş Genişletme: Çalışanlar her ne kadar belirli bir iş ve görev grubunda uzmanlaşmış olurlarsa olsunlar yeni iş ve uzmanlıklar için yeni beceri ve yetenekler geliştirebilmektedirler. • Düşük uzmanlaşma: Çalışanların belirli görevlerde uzmanlaşması yerine örgütün genel hedeflerine göre sürekli değişen ve gelişen iş süreçleri mevcuttur. • Düşük formalleşme: Yazılı kurallar olmadığından görev ve sorumluluklar her an güncellenebilir. • Düşük formalleşme: Yazılı kurallar olmadığından görev ve sorumluluklar her an güncellenebilir. • İşe bağlılık: Çalışanlar arasında işe bağlılık yaygındır, bu tip örgütlerde işe bağlılık itaat ve sadakatten daha değerlidir. • İşe bağlılık: Çalışanlar arasında işe bağlılık yaygındır, bu tip örgütlerde işe bağlılık itaat ve sadakatten daha değerlidir. • Adem-i Merkeziyetçilik: Kararlar bilgisi olanlar tarafından verilir. Kontrol, yetki ve iletişim örgüt içi ağ yapısı ile düzenlenir. • Adem-i Merkeziyetçilik: Kararlar bilgisi olanlar tarafından verilir. Kontrol, yetki ve iletişim örgüt içi ağ yapısı ile düzenlenir. • Kararlara katılım: Bilgi örgütte kimsenin tekelinde değildir, teknik ya da ticari bilgi örgütün herhangi bir kademesinde bulunabilir, yetki ve iletişimin temel belirleyicisidir. • Yatay iletişim ve koordinasyon: Departmanlar arası iletişimi destekleyen yatay bir iletişim yapısı vardır. • Düşük hiyerarşi ve örgütsel hedefler: Bireysel hedeflerden çok örgütsel hedeflerin gerçekleştirilmesi önemlidir. İletişimin içeriği kural ve kararlardan çok bilgi ve tavsiyelere dayanmaktadır. • Çevresel bilgi: örgüt faaliyet gösterdiği çevre içindeki ticari, ekonomik, yasal, demografik gelişmelerden haberdardır. Çevresel değişimlerin farkında olmak ve bu değişimlere ait bilgiye sahip olmak oldukça önemlidir. 3.2.2 Organik Örgüt Yapısı ve İnsan Kaynakları Yapılanması Organik örgütün özelliklerine bakıldığında şeffaflık, bilginin paylaşımı, katılım, esneklik, çeviklik ve kararlarda çalışanların öncelik kullanabilme becerilerinin öne çıktığı görülmektedir. Dolayısıyla örgütsel stratejilerle IK süreçlerinin bütünleşme ve uyum aşamasında organik bir işletme tasarımının örgütsel performans konusunda daha fazla katkı sunması öngörülebilir. Fazilet Nohut / Özlem Balaban \| 217 217 Fazilet Nohut / Özlem Balaban \| 217 SONUÇ Bir örgütün stratejik başarısının belirleyicisi örgütün fonksiyonları, yönetim şekli, yöneticilerin yetki ve sorumlulukları, liderlik tarzları vb. yani kısacası örgütün yapısı ile stratejisinin arasındaki uyumdur. Örgütün belirlemiş olduğu stratejiyi geliştirmeye uygun olmayan yapılar, örgüt içi çatışmaların, yüksek maliyetin, düşük performansın ve örgütsel başarısızlığın kaynağını oluştururlar. Dolayısıyla öncelikle başarılı stratejik karar ve uygulamalara zemin hazırlayacak bir örgüt kültürünün oluşturulması son derece önemlidir. Drucker (2016)’nın söylediği gibi “kültür stratejiyi kahvaltı niyetine yer” mi bilinmez ama stratejinin her zaman başarıyı garantilemediği (Coşkun, 2016) bir gerçektir. Bu bağlamda örgütün vizyonunun, misyonunun, değerlerinin belirlenmesi ve bu değerlerin çalışanlar tarafından benimsenmesinin sağlanması önemlidir. Mevcut ve gelecekteki çalışanların örgüt kültürüne uyumlu olması, amaç, değer ve hedeflerini benimsemesi örgütün bir bütün olarak aynı istikamette hareket etmesini kolaylaştıracaktır (Dinçer, 2004). Diğer taraftan örgütte var olan liderlik tarzı da stratejik başarının sağlanmasında son derece önemlidir. Bugünün hızla değişen ve dönüşen dünyasında çevik yönetim uygulamalarını, esnek yapıları ve hızlı karar alma becerilerini zorunlu kılmaktadır. Dahası stratejik yönetimin önemli bir enstürümanı olan uzun vadeli planların bile artık anlamını ve önemini yitirdiği bir çağ deneyimlenmektedir. Dolayısıyla geleneksel liderlik tarzlarıyla çağın getirdiği bu dinamizmin gerekliliklerinin sağlanması pek mümkün görünmemektedir. Bu nedenle liderlik tarzlarından iş yapış biçimlerine kadar iş tasarımlarının gözden geçirilmesi, çevresel değişim ve dinamizme adapte edilebilmesi gerekmektedir. Çalışan beklentilerindeki değişimler, kariyer kavramının ve anlamının geleneksel bakış açısından uzaklaşması, iş yapma biçimlerindeki değişimlerle birlikte yeni tanımlamaların (uzaktan çalışma, hibrit çalışma, dijital göçebeler vb.) gündeme gelmesi iş süreçlerinin örgütsel stratejiyi gerçekleştirecek şekilde yeniden dizayn edilmesini zorunlu kılmaktadır. Bu araştırmanın teorik arka planının argümanları kaynak temelli yaklaşım ve adaptasyon kuramlarına dayanmaktadır. Strateji kavramı ve İKY süreçlerinin uyumunun önemi ise Porter’ın rekabet stratejileri ve IK yapılanması çerçevesinde incelenmiştir. Bu stratejilerin toplam maliyet liderliği, farklılaşma ve odaklanma olarak ifade edilen ve her biri benimsendiğinde örgütün içi ve dış çevresine yönelik uygulamalarda bir takım belli rol ve davranışları gerekli kılmaktadır. Ayrıca örgütler zamanla büyüme, küçülme, tasarruf stratejileri gibi büyüklüğü ile ilgili kararlar veya savunmacı, araştırmacı ve analizci gibi örgütün tepkilerini nasıl vereceğinin temelini belirleyen stratejiler benimseyebilmektedir. Benimsenen ve uygulanan 218 Rekabet Stratejileri Bağlamında Stratejik İnsan Kaynakları Yönetiminin Rolü ve Önemi bu stratejilerin örgütün tüm fonksiyonlarında karşılık bulması stratejinin daha sağlam bir zeminde hayata geçirilmesine ve çalışanlar tarafından içselleştirilmesine olanak tanımaktadır. Dolayısıyla İKY’nin politika ve uygulamalarının da örgüt stratejisiyle senkronize biçimde hareket etmesinin örgütün performansına olumlu katkılar sağlayacağı düşünülmektedir. Stratejik İKY olarak ifade edilen bu anlayışta “uyum” kavramı merkezde yer almaktadır. SONUÇ Uyumdan kasıt hem örgüt içerisinde İKY fonksiyonlarının örgüt stratejisiyle entegrasyonu, hem de örgüt dışında örgütün stratejisi ile İKY uygulamalarının entegrasyonu ifade edilmektedir. Bu amaçla her bir fonksiyon açısından örgütün hangi gelişim aşamasında olduğunun farkında olunmalıdır. Bu doğrultuda gerçekleştirilen stratejik adımların nasıl sonuçlara sebep olabileceği bilinciyle belirli davranışlar yapılandırılmalıdır. 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Üye, Sinop Üniversitesi, MYO, Muhasebe ve Vergi Bölümü, uarslan@sinop.edu.tr, orcid: 0000-0001-7395-8576 KAYNAKÇA Strategic human resource ma- nagement and its effects on firm performance: An implementation of the competing values framework, The international human resource manage- ment, 14(4), 680-699. Pearce II, John A., Robinson,Jr., B.(2015), Stratejik yönetim, geliştirme, uygula- ma ve kontrol, McGrawHill Irwin, Çeviri Editörü: Prof.Dr. Mehmet Bar- ca, On ikinci basımdan çeviri, Nobel yayın, Ankara Porter, M. (1980). Competitive strategy: Techniques for snalysing industries and competitors. New York: The Free Press. Porter, M. (1985). Competitive advantage: Creating and sustaining superior perfor- mance. New York: The Free Press. Schuler, R. S., Jackson, S. E. (1987). Linking competitive strategies with hu- man resource management practices. The academy of management execu- tive, 1(3), 207–219. Sheppeck, M. A., & Militello, J. (2000). 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Dr., Bandırma Onyedi Eylül Üniversitesi, İ.İ.B.F., İşletme Bölümü, metinkilic@bandirma.edu.tr, orcid:0000- 0002-5025-6384 1 Dr. Öğr. Üye, Sinop Üniversitesi, MYO, Muhasebe ve Vergi Bölümü, uarslan@sinop.edu.tr, orcid: 0000-0001-7395-8576 2 Doç. Dr., Bandırma Onyedi Eylül Üniversitesi, İ.İ.B.F., İşletme Bölümü, metinkilic@bandirma edu tr orcid:0000- 0002-5025-6384 1. Giriş TÜRMOB’un yapmış olduğu bu çalışmalar, konuya verdiği önemi göstermesinin yanında sorun çözüme kavuşuncaya kadar mücadeleden vazgeçmeyeceğini de göstermektedir. TÜRMOB’un yapmış olduğu bu çalışmalar, konuya verdiği önemi göstermesinin yanında sorun çözüme kavuşuncaya kadar mücadeleden vazgeçmeyeceğini de göstermektedir. Meslekteki haksız rekabet sorununu ortadan kaldırmak amacıyla yapılan bu çalışmaların sorunun çözümündeki etkisini tespit etmek ve haksız rekabet dosyalarının içerik yönünden zaman içinde değişiklik gösterip göstermediğinden hareketle bu konuda genellemelere varmayı amaçlayan araştırmada, Ekergil (2018)’in 2008-2016 yılları arasındaki haksız rekabet dosyalarını analiz ettiği çalışmasına benzer yöntemlerle 2017-2022 yılları arasındaki haksız rekabet dosyaları analiz edilmiştir. Araştırmada, 2016 yılı sonrası döneme ilişkin analizler yapılmış ve bu sonuçlar 2016 yılı öncesi döneme ilişkin çalışmanın sonuçlarıyla karşılaştırılarak aradaki mevcut farklılıklar ifade edilmiştir. Ayrıca her iki dönem birlikte değerlendirilip 2008-20022 dönemine ilişkin genellemeler yapılmıştır. Araştırmada öncelikle haksız rekabet durumlarında muhasebe meslek mensuplarına uygulanacak idari yaptırım süreci açıklanmıştır. TÜRMOB tarafından düzenlenerek meslek camiasının bilgisine sunulan bu süreçte, SMMM odaları ve YMM odalarından oluşan muhasebe meslek odalarında Oda Yönetim Kurullarının (OYK), Oda Haksız Rekabetle Mücadele Kurullarının (OHRMK) ve Oda Disiplin Kurullarının (ODK) yapmış olduğu çalışmalar açıklanmıştır. Böylece haksız rekabet yaptırım sürecinde ifade edilen araştırma verilerinin, okuyucular tarafından daha kolay anlaşılabilir olmasına çalışılmıştır. Daha sonra araştırılan konuya ilişkin literatür taramasına yer verilmiştir. Son bölümde ise araştırma bulgularına yer verilerek haksız rekabet dosyalarının analizi gerçekleştirilmiştir. 1. Giriş Genel anlamda rekabet hakkının kötüye kullanılması olarak ifade edilebilen haksız rekabet kavramı muhasebe mesleğinde, Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Müşavirlik Mesleklerine İlişkin Haksız Rekabet ve Reklam Yasağı Yönetmeliği’nin (HRRYY) tanımlar başlığını taşıyan dördüncü maddesinde; “Meslek mensuplarının birbirleriyle ve iş sahibiyle https://doi.org/10.58830/ozgur.pub401.c1761 https://doi.org/10.58830/ozgur.pub401.c1761 221 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 222 olan ilişkilerini etkileyen aldatıcı veya diğer şekillerdeki dürüstlük kurallarına aykırı davranışları veya mesleki uygulamalar” biçiminde tanımlanmıştır. Aynı yönetmeliğin beşinci maddesinde “Meslek mensupları, mesleki faaliyetleri kapsamında işin kabulü, reddi, yürütülmesi ve sona ermesiyle reklam yasağı kapsamına giren her türlü faaliyetlerine ilişkin olarak haksız rekabete neden olacak eylem ve davranışlarda bulunamazlar” hükmüne yer verilerek muhasebe meslek mensuplarının haksız rekabet davranışında bulunmamaları gerektiği vurgulanmıştır. Muhasebe mesleğinde haksız rekabet halleri, HRRYY’de 3 madde ve bu maddeler altında toplanan 27 bentte ifade edilmiştir. Bunlar, yönetmeliğin 6. maddesinde; “Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet” başlığı altında 11 bent, 7. maddesinde; “Ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet” başlığı altında 9 bent ve 8. maddesinde; “Reklam yoluyla haksız rekabet” başlığı altında 7 bentten oluşmaktadır. HRRYY’de belirtilen bu haksız rekabet hallerinden birinin ihlali durumunda ihlali gerçekleştiren meslek mensubu hakkında idari disiplin süreci başlatılarak, meslek için düzenlenmiş disiplin yönetmeliğine göre kendisine yaptırım uygulanmaktadır. Meslek mensubuna uygulan idari yaptırım süreci araştırmanın ikinci bölümünde açıklanmıştır. Serbest Muhasebeci Mali Müşavirler (SMMM) Odaları ile Yeminli Mali Müşavirler (YMM) Odalarının üst birliği olan ve haksız rekabet sorununun çözüm otoritesi olarak ifade edebileceğimiz, Türkiye Serbest Muhasebeci Mali Müşavirler ve Yeminli Mali Müşavirler Odaları Birliği (TÜRMOB) sorunla mücadele kapsamında günümüze kadar pek çok düzenleme ve uygulama gerçekleştirmiştir. TÜRMOB’un sorunun çözümüne yönelik yapmış olduğu bu çalışmaları aşağıdaki başlıklar halinde özetleyebiliriz. • 03.01.1990 tarihinde “Serbest Muhasebeci Mali Müşavir ve Yeminli Mali Müşavirlerin Çalışma Usul ve Esasları Hakkında Yönetmelik” yayımlanmıştır. • 21.11.2007 tarihinde “Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Müşavirlik Mesleklerine İlişkin Haksız Rekabet ve Reklam Yasağı Yönetmeliği” yayımlanmıştır. • TÜRMOB bünyesinde, amacı meslekteki haksız rekabet sorunuyla mücadele olan “Türkiye Haksız Rekabetle Mücadele Kurulu (TÜRHAK)” oluşturulmuştur. • TÜRHAK tarafından çok sayıda “bilgilendirme ve yayım çalışması” gerçekleştirilmiştir. 223 Ünal Arslan / Metin Kılıç • İlki 2009 yılında olmak üzere sekiz adet “haksız rekabet kongresi” düzenlenmiştir. • 2012 yılı “haksız rekabetle mücadele yılı” ilan edilmiştir. • 2012 yılı “haksız rekabetle mücadele yılı” ilan edilmiştir. • “Haksız Rekabetle Mücadele Yazılımı (E-Birlik Projesi)” hazırlanmış ve meslek mensuplarının kullanımına sunulmuştur. • “Haksız Rekabetle Mücadele Yazılımı (E-Birlik Projesi)” hazırlanmış ve meslek mensuplarının kullanımına sunulmuştur. 2. Haksız Rekabet Durumlarında Oda Kurullarının Yaptırım Sürecindeki Faaliyetleri Muhasebe mesleğinde haksız rekabet olarak ifade edilen durumların ortaya çıkması halinde bu davranışlarda bulunan meslek mensuplarına, meslek için düzenlenmiş disiplin yönetmeliğine göre yaptırım uygulamak OYK’nın görevidir. OYK bu görevini, OHRMK ve ODK ile koordineli bir biçimde yerine getirmektedir. Bu kurullar haksız rekabet davranışının oluşması ve bu 224 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi oluşuma sebebiyet veren meslek mensubuna yaptırım uygulanması sürecinde Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Müşavirlik Kanunu Disiplin Yönetmeliği, HRRYY ve Odalar Arası Uygulama Birlikteliği Raporuna (OAUBR) göre çalışmalarını sürdürmektedir. Bu süreçte adı geçen oda kurullarının yapmış olduğu çalışmalar aşağıda ifade edilmiştir (Arslan, 2020:106). 2.1. Oda Yönetim Kurulunun Yaptırım Sürecindeki Faaliyetleri Haksız rekabetle ilgili disiplin süreci soruşturma ve kovuşturma olmak üzere iki aşamadan oluşmaktadır. Soruşturma aşaması; ihbar veya şikâyetin odaya ulaşmasından dosyanın disiplin kuruluna sevk edilmesine kadar geçen aşamayı, kovuşturma aşaması ise; dosyanın disiplin kuruluna intikalinden disiplin kurulu kararının kesinleşmesine kadar olan aşamayı ifade etmektedir (OAUBR, 2011:35; Arslan, 2020:106). Haksız rekabet konusunda soruşturma sürecinin ilk aşamasında “soruşturmayı başlatan olay” OYK’ya bildirilir. Bu noktada soruşturmayı başlatan olay OAUBR’da; ilgilinin ihbar veya şikâyeti, oda ve birlik kurullarından herhangi birinin isteği ve hükümlülükle biten bir yargılama sonucu verilen mahkeme kararının odaya intikali olmak üzere üç başlık halinde ifade edilmiştir. Bunlardan ilki olan ilgilinin ihbar veya şikâyeti; haksız rekabet davranışına maruz kaldığını iddia eden herhangi bir gerçek veya tüzel kişinin oda veya birlik kurullarına yazılı veya sözlü şekilde başvurarak, hakkında ihbar veya şikâyette bulunduğu meslek mensubunu belirtip iddialarını açıklamasıyla yapılmış sayılmaktadır. Odaya yapılan ihbar veya şikayetlerin yazılı şekilde yapılması gerekir. Bu yüzden sözlü şikâyet veya ihbarların odada mutlaka tutanağa bağlanarak, tutanağın ihbar veya şikâyette bulunan kişiyle en az bir oda kurul üyesi tarafından imzalanması gerekmektedir. Oda ve birlik kurullarından herhangi birinin gerekli gördüğü durumlarda soruşturma açma isteği soruşturmayı başlatan olaylardan ikincisini oluşturmaktadır. Bünyelerinde mesleki denetim komitesi, büro denetleme komitesi vb. adlarla oluşturulmuş birimler bulunan odalar, bu birimlerce yapılan denetimler sonucunda düzenlemiş oldukları tutanaklara istinaden re’sen soruşturma açma hakkına sahiptirler. Soruşturmayı başlatan olay olarak nitelendireceğimiz üçüncü durum ise; meslek mensubunun yargılanması sonucu hükümlülükle biten bir mahkeme kararının odaya iletilmesidir. OYK; sonucu kendilerine iletilmiş olması şartıyla hükümlülükle sonuçlanmış bir ceza davasının konusunu oluşturan eylemden dolayı ayrıca disiplin soruşturması açmak zorundadır (OAUBR, 2011:36-37; Arslan, 2020:106-107). 225 Ünal Arslan / Metin Kılıç OYK ihbar veya şikâyetin odaya intikalinden itibaren en geç iki ay içerisinde ihbar veya şikâyeti incelemek zorundadır (OAUBR, 2011:38; Arslan, 2020:107). OYK şikâyet veya ihbar olayının; meslek mevzuatına uygunluğunu, disiplin yönetmeliğine göre yaptırıma bağlanabilirliğini ve ceza davasına konu olup olmadığını ilgili toplantıda karara bağlamaktadır. OYK yapmış olduğu bu incelemede söz konusu ihbar veya şikâyet meslek mevzuatından kaynaklanmıyor ve disiplin yönetmeliği uyarınca da bir yaptırıma bağlanmıyor ise olay hakkında “soruşturma açılmasına yer olmadığına” karar verebilir. Aksi durumda ise şikâyet veya ihbar konusunda soruşturma kararı almaktadır. OYK soruşturma kararı alırken aynı zamanda şikâyet veya ihbar konusunun hangi yönetmeliğe aykırı bir davranış oluşturduğunu da tespit etmektedir. Görüşülen konu; haksız rekabetle veya reklam yasağıyla ilgiliyse OHRMK’ya, disiplin cezasıyla ilgili bir konu ise ODK’ya dosyanın sevki gerçekleştirilir. 2.1. Oda Yönetim Kurulunun Yaptırım Sürecindeki Faaliyetleri Ayrıca HRRYY’nin 32/1 maddesinde “Bu yönetmelik hükümlerine aykırılık tespit edilmesi halinde, odalar tarafından re’sen veya şikâyet üzerine disiplin soruşturması başlatılır” hükmü yer almaktadır. Bu hükümle OYK, kendisine gelen haksız rekabet dosyasının bir disiplin suçu gerektirdiği kanaatine varırsa, söz konusu dosyayı OHRMK’ya sevk etmeden doğrudan ODK’ya gönderebilme hakkına da sahiptir (Ekergil, 2018:58; Arslan, 2020:107). 2.2. Oda Haksız Rekabetle Mücadele Kurulunun Yaptırım Sürecindeki Faaliyetleri Odalarda yürütülen haksız rekabet yaptırım sürecini HRRYY hükümleri net bir şekilde ortaya koyamamaktadır. Öyle ki HRRYY’de OHRMK’nın OYK’dan almış olduğu soruşturma yapma yetkisini nasıl gerçekleştireceğine dair bir bilgi mevcut değildir. Bu eksiklik, OAUBR ile giderilerek haksız rekabet soruşturma süreci hakkında ilgililere bilgi sağlanmıştır. Sürecin birçok aşamasında “Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Müşavirlik Kanunu Disiplin Yönetmeliği’nin” hükümleri esas alınmaktadır. Buna göre OHRMK’larda soruşturma süreci disiplin yönetmeliğinin 19. maddesindeki hükümlere göre gerçekleştirilmektedir (Ekergil, 2018:54-60; Arslan, 2020:107-108). Bu süreçte OHRMK’lara düşen görev, hakkında haksız rekabet suçu işlediğine dair iddia ile soruşturma açılan meslek mensubunun soruşturmasını gerçekleştirmek ve bu soruşturma sonrasında görüşünü bir rapor halinde OYK’ya sunmaktır. Bu göreviyle ilgili olarak OHRMK, hakkında ihbar veya şikâyette bulunulan meslek mensubuyla ihbar veya şikayette bulunan kişiyi dinlemekte, hakkında soruşturma yürütülen meslek mensubunu mutlaka görüşmeye davet etmekte ve soruşturma konusu iddialarla ilgili olarak Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 226 açıklamalarını tutanakla tespit etmektedir. Ancak hakkında soruşturma yürütülen meslek mensubunun görüşmeye davet edilmesi, savunma alma ile karıştırılmamalıdır. Bu aşamada, soruşturmacı tarafından yapılan işlem savunma alma değil, bilgi alma amaçlı bir işlemdir. Savunma alma ODK’nın görevidir. OHRMK’da soruşturmayla görevlendirilmiş kişi veya kişiler kanıtları toplar, isterse meslek mensubunun bürosunda denetim yapabileceği gibi bu konuda oda bünyesinde oluşturulmuş komitelerden denetim yapılmasını OYK aracılığıyla isteyebilir. Ayrıca soruşturmacının gerekli gördüğü bilgi ve belgeleri resmi veya özel kuruluşlardan OYK’nın aracılığıyla talep etme hakkı da bulunmaktadır (OAUBR, 2011:39-40; Arslan, 2020:108). OHRMK yapmış olduğu soruşturma sonucunda, meslek mensuplarının Haksız Rekabet ve Reklam Yasağı Yönetmeliği’ne aykırı hareketlerinin bulunmadığını tespit ederse “işlem yapılmasına gerek olmadığı” görüşüyle, aksi durumda ise soruşturmayla ilgili yaptırım önerisini de belirttiği disiplin kovuşturması açılması kanaatini taşıyan inceleme raporunu OYK’ya bildirir. OHRMK yapmış olduğu soruşturma sonucu, meslek mensubuna uygulanacak yaptırım hakkında OYK’ya sadece kanaat (görüş) bildirme yetkisine sahiptir, bunun dışında herhangi bir karar verme hak ve yetkisi yoktur (OAUBR, 2011:9; Arslan, 2020:108). 2.3. Oda Disiplin Kurulunun Yaptırım Sürecindeki Faaliyetleri OHRMK’dan disiplin kovuşturması açılması isteğiyle OYK’ya gelen dosya OYK tarafından değerlendirilir. OYK bu durumda iki şekilde davranabilir. Birincisi, kovuşturma açılmasına yer olmadığına ilişkin bir karar verebilir ki bu noktada bu karara OHRMK’nın itiraz hakkı bulunmamaktadır. Bu kararın ilgililere tebliğ edilmesinden itibaren 30 gün içinde şikayetçi meslek mensubu tarafından oda aracılığıyla yahut doğrudan Birlik Disiplin Kurulu’na itiraz edilebilir. İkinci durum ise OYK’nın OHRMK’nın görüşü doğrultusunda ilgili meslek mensubu hakkında kovuşturma açılması yönünde bir karar verip dosyayı ODK’ya sevk etmesidir. Bu durum, haksız rekabet suçu işlediği iddiasıyla hakkında soruşturma açılan meslek mensubu için soruşturma aşamasının tamamlanıp kovuşturma aşamasına geçildiğini ifade etmektedir (Arslan, 2020:108-109). Kovuşturma aşamasında; ODK kendisine gelen dosyayı OYK’ca kovuşturma açılan eylemle sınırlı olarak inceler. Bir başka deyişle, ODK kovuşturmayı yürütürken kovuşturma konusu eylem dışında meslek mensubunun mevzuata uygun olmayan başka eylemlerinin olduğunu tespit ederse, bu eylemleri yürütmüş olduğu kovuşturmaya dahil edemez. Bu 227 Ünal Arslan / Metin Kılıç \| durumda ODK’ca yapılması gereken işlem, tespit edilen eylemi OYK’ya bildirmektir. Ayrıca ODK incelediği dosya hakkında karar verirken, OYK’nın eylem hakkındaki nitelemesiyle bağlı değildir. Şöyle ki, ODK karar verirken OYK’ca önerilen ceza maddesi dışında başka bir hükümden ceza verebilir veya isnat edilen suçun oluşmadığı gerekçesiyle ceza verilmesine yer olmadığına karar verebilir (OAUBR, 2011:44; Arslan, 2020:109). ODK, dosyanın kendisine ulaşmasından sonra ilk olarak dosyadaki delilleri inceler. Gerek görürse, ek tespitlerin yapılmasını veya ilgili kuruluşlardan bilgi ve belge teminini OYK’dan talep eder. ODK’nın bu süreçte bilirkişi incelemesi yaptırma ve tanık dinleme yetkisi de bulunmakla birlikte sürecin en önemli konusu haksız rekabet davranışında bulunduğu iddia edilen meslek mensubunun savunmasının alınmasıdır. Anayasanın 129. maddesi ve 3568 sayılı Kanun’un 48. maddesi gereğince meslek mensuplarının savunması alınmadan onlara disiplin cezası verilemez. Meslek mensuplarına tebliğ edilen savunma istem yazılarında kendilerine hangi eylemlerinden dolayı kovuşturma açıldığının açıkça bildirilmesi gerekirken, savunma istem yazısının kendilerine tebliğinden itibaren yazılı savunmalarını disiplin kuruluna sunmaları için en az 15 günlük süre tanınması da zorunludur (OAUBR, 2011:44-45; Arslan, 2020:109). ODK, hakkında haksız rekabet davranışında bulunduğu iddia edilen meslek mensubunun savunmasını aldıktan, talep üzerine veya re’sen gerek gördüğü taktirde duruşma yaptıktan, ilgili kişileri dinledikten ve yapılacak tüm incelemeleri tamamladıktan sonra kararını verir. Bu kararın, ihbar ve şikâyetten başlayıp soruşturma ve kovuşturma aşamasında elde edilen kanıtları, ilgili kişinin savunmasını, ilgili mevzuatı, bu delillerin değerlendirilmesini ve bu değerlendirmeler sonunda mevzuata aykırılık tespit edilmesi halinde eylemin disiplin yönetmeliğinin hangi hükmünün ihlalinden oluştuğunu belirtir biçimde kaleme alınması gerekmektedir. 2.3. Oda Disiplin Kurulunun Yaptırım Sürecindeki Faaliyetleri ODK, meslek mensubunun hakkında iddia edilen haksız rekabet davranışında bulunduğuna kanaat getirirse ceza, haksız rekabet davranışının oluşmadığına kanaat getirirse ceza verilmesine gerek olmadığına dair karar verir (OAUBR, 2011:47-48; Arslan, 2020:110). 4.1. Araştırmanın Amacı ve Yöntemi Araştırmanın amacı; meslekteki haksız rekabet sorununun çözümü amacıyla yapılan çalışmaların, sorunun çözümündeki etkisini tespit etmek ve haksız rekabet dosyalarının içerik yönünden yıllar itibarıyla bir değişiklik gösterip göstermediğinden hareketle araştırılan döneme ilişkin genellemelere varmaktır. Bu amaçla, 2016 yılı sonrası döneme ait haksız rekabet dosyaları sekiz başlık altında analize tabi tutulmuş, sonuçlar 2016 yılı öncesi dönem sonuçlarıyla karşılaştırılmış ve iki dönem arasındaki farklılıklar ifade edilmiştir. Ayrıca 2008-2022 dönemi bir bütün olarak değerlendirilerek bu döneme ilişkin bazı genellemelere varılmıştır. Araştırmada, betimleyici araştırma yöntemi kullanılmıştır. Betimleyici araştırmalar, ilgi duyulan konunun betimlemesini, tasvirini elde etmeyi amaçlayan araştırmalardır ki bu araştırmalarda çalışılan olgu ya da örneklem hakkında elde edilen veriler betimlenerek temel özellikleri tasvir edilmektedir (Aslan ve Özdemir, 2020:167). Bu kapsamda araştırmada, muhasebe mesleğinde haksız rekabet sorununa ilişkin bir durum tespiti yapılmış ve bu tespit sonuçlarına ilişkin değerlendirmelerde bulunulmuştur. Araştırma verileri, TÜRMOB’un resmi web sayfasında 2008-2022 dönemini kapsayan faaliyet raporlarında yer alan TÜRHAK raporlarından elde edilmiştir. Elde edilen bu veri seti Microsoft Office Excel programında analiz edilmiştir. 5. Araştırma Bulguları 3. Literatür Literatür incelendiğinde muhasebe mesleğinde haksız rekabet dosyalarının analizini konu alan çalışma olarak sadece Ekergil (2018)’in çalışmasına ulaşılabilmiştir. Bu çalışmayla birlikte, meslekteki haksız rekabet sorununa yönelik olarak hazırlanmış olup içeriği bakımından araştırma konusuyla ilişkilendirebileceğimiz çalışmalar aşağıda ifade edilmiştir. 228 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi Ömürbek ve Türkoğlu (2013) çalışmalarında, meslekteki haksız rekabet sorununu ve bu soruna neden olan etkenleri meslek mensuplarının mesleki deneyim sürelerine göre belirlemeyi amaçlamışlardır. Çalışmalarında, meslekte ücret üzerinden haksız rekabet yaşandığı ve meslek mensuplarının meslekleri dışında işlerle uğraşmalarının haksız rekabetin etkisini daha da artırdığı sonuçlarına ulaşmışlardır. Sak (2013) çalışmasında, haksız rekabet sorununun tespitini ve bu soruna ilişkin çözüm önerileri geliştirmeyi amaçlamıştır. Çalışmasında, kaçak bürolar, ruhsat kiralama, asgari ücret tarifesine uyulmama ve her türlü muhasebe işini üstlenen denetim firmalarını meslekte haksız rekabete yol açan önemli unsurlar olarak ifade etmiştir. Ayrıca çalışmasında muhasebe meslek odalarının sorunun çözümünde gerekli etkiyi sağlayamadığını vurgulamıştır. Akarsu (2015) çalışmasında, muhasebe meslek mensuplarının meslekteki haksız rekabet sorununa ilişkin düşüncelerini, bilgilerini ve mevcut algılarını öğrenmeyi amaçlamıştır. Çalışmasında, meslek mensupları arasında kısmen de olsa haksız rekabet sorununun mevcut olduğu ayrıca meslekte zaman zaman ruhsat kiralama durumlarının yaşandığı, mesleki standartlara kısmen uyulduğu, sözleşme değerinin altında serbest meslek makbuzu düzenlendiği, asgari ücret tarifesinin altında ücret talep eden meslek mensuplarının olduğu, iş alabilmek adına bir takım vaatlerde bulunulduğu, meslektaşları hakkında yanıltıcı açıklamalar yapan ve gerçek dışı bilgiler veren meslek mensuplarının olduğu sonuçlarına ulaşmıştır. Ekergil (2018) çalışmasında, muhasebe mesleğinde haksız rekabet- disiplin ilişkisini açıklamış, haksız rekabet durumlarında meslek mensuplarına uygulanacak yaptırım süreci ve yaptırım türlerini belirtmiştir. Ayrıca çalışmasında 2008-2016 yılları arasında muhasebe mesleğindeki haksız rekabet dosyalarını analiz etmiştir. Bu analizini TÜRHAK raporlarında belirtilen başlıklar altında gerçekleştirmiştir. Karakaya ve Biçer (2020) muhasebe meslek mensuplarının haksız rekabet algılarının ölçülmesini amaçladıkları araştırmalarında, meslek mensuplarıyla yaptıkları görüşmeleri analitik hiyerarşi yöntemiyle değerlendirmişlerdir. Çalışmalarında, haksız rekabete konu olan etkenleri önem derecelerine göre yüzdelik değerlerle şu şekilde sıralamışlardır; mesleki faaliyetlerinde özen göstermemek (%22), ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet (%15), muhasebe ve denetim standartlarına uymamak (%13), meslek ruhsatının kiralanması (%12), reklam yoluyla haksız rekabet (%10), mevzuat hükümlerine aykırı davranmak (%9), izinsiz faaliyette bulunmak (%8), gerçeğe aykırı belge düzenlemek ve onaylamak (%7), mesleki ve akademik unvan dışında sahip olunan unvanları kullanmak (%6). Ünal Arslan / Metin Kılıç \| 229 4.2. Araştırmanın Sınırlılıkları Araştırma verilerinin sadece 2008-2022 dönemine ait verileri kapsaması araştırmanın sınırlılığını oluşturmaktadır. 2008 yılı öncesi döneme ait verilere ulaşılamamasının sebebi; TÜRHAK’ın, HRRYY’nin 24. maddesi uyarınca 2008 yılında oluşturulması ve o yıldan itibaren haksız rekabete ilişkin verileri yayımlamaya başlamasıdır. Araştırmada, TÜRHAK raporlarında yer alan tüm veriler analize dahil edilmiş olup sadece bu verilerin ayrıntılı gösterimi olan haksız rekabet dosyalarının Türkiye dağılımı kapsam dışı bırakılmıştır. 5. Araştırma Bulguları 5.1. Oda Yönetim Kurulu Kararıyla Oda Haksız Rekabetle Mücadele Kurullarına Sevk Edilen Dosya Sayısı 5.1. Oda Yönetim Kurulu Kararıyla Oda Haksız Rekabetle Mücadele Kurullarına Sevk Edilen Dosya Sayısı 2008-2022 yılları arasında OYK’lar tarafından OHRMK’lara sevk edilen haksız rekabet dosya sayıları Grafik 1’de sunulmuştur. Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 230 Grafik 1: OYK’lar Tarafından OHRMK’lara Sevk Edilen Dosya Sayısı Grafik 1: OYK’lar Tarafından OHRMK’lara Sevk Edilen Dosya Sayısı OYK kararıyla OHRMK’lara sevk edilen dosya sayısında 2017-2022 yılları arasında artış olduğu görülmektedir. 2021 yılında bir önceki yıla göre bir miktar azalış olsa da 2022 yılında dosya sayısının tekrar artışa geçtiği görülmektedir. 2021 yılında dosya sayısındaki azalışın COVİD-19 salgını kaynaklı olduğu düşünülmektedir. 2016 yılı öncesi dönemde durağan seyreden ve en fazla 1.349 sayısına ulaşan haksız rekabet dosya sayısı 2016 yılından itibaren artış trendiyle 2020 yılında 3.958 ve 2022 yılında 4.379 dosya sayısıyla incelenen dönemdeki maksimum düzeye ulaşmıştır. 2008- 2022 dönemi incelendiğinde, OYK kararı ile OHRMK’lara sevk edilen dosya sayısında belirgin bir artış olduğu görülmektedir. Haksız rekabet dosya sayısındaki bu artış, sorununun çözümü için yapılan çalışmaların istenilen başarıyı sağlayamadığını göstermektedir. 5.2. Oda Haksız Rekabetle Mücadele Kurullarının Oda Yönetim Kurullarına Sunduğu Dosyaların Dağılımı OHRMK’lardan OYK’lara gönderilen dosyalar dört başlık altında sınıflandırılmaktadır. OHRMK dosya üzerinde yapmış olduğu incelemeler sonucunda dosyada haksız rekabet suçunun işlendiği kararını verirse OYK’ya “yaptırım önerisiyle (YÖ)” dosyayı sunmakta, yapmış olduğu incelemeler sonucunda haksız rekabet suçunun işlenmediği kararını verirse “ceza verilmesine yer olmadığı önerisiyle (CVYO)” OYK’ya raporunu sunmaktadır. Hakkında ihbar, şikâyet veya istemde bulunulan meslek mensubu soruşturma sürecinde akıl sağlığını sürekli kaybetmiş veya ölmüşse, OYK tarafından “dosyanın işlemden kaldırılması (DİK)” kararı verilir. Bu şartlar oluştuğunda meslek mensubu hakkındaki soruşturma sonlandırılır ve bu dosyalar dosyanın 231 Ünal Arslan / Metin Kılıç işlemden kaldırılması başlığı altında toplanır. “Sonuçlandırılmamış dosyalar (SD)”; OHRMK’larca ilgili dönemde sonuçlandırılamadığı için bir sonraki dönemde sonuca bağlanacak dosyaları ifade etmektedir (Ekergil, 2018:90- 94). 2008-2022 yılları arasında OHRMK’ların OYK’lara gönderdiği dosyaların dağılımı Grafik 2’de sunulmuştur. Grafik 2: OHRMK’ların OYK’lara Sunduğu Dosyaların Dağılımı Grafik 2: OHRMK’ların OYK’lara Sunduğu Dosyaların Dağılımı Grafik 2: OHRMK’ların OYK’lara Sunduğu Dosyaların Dağılımı OHRMK’larca OYK’lara sunulan dosyaların dağılımı incelediğinde; 2016 yılı sonrasında, ceza verilmesine yer olmadığı önerisiyle ve dosyanın işlemden kaldırılması önerisiyle OYK’ya sunulan dosya sayılarında belirgin bir değişme olmazken, sonuçlandırılmamış dosya sayısında ve yaptırım önerisiyle OYK’ya sunulan dosya sayısında önemli bir artış olduğu görülmektedir. Özellikle 2019 yılı ve sonrasında sonuçlandırılmamış dosya sayısındaki artış dikkat çekmektedir. 2008-2022 dönemi incelendiğinde; ceza verilmesine yer olmadığı önerisiyle ve dosyanın işlemden kaldırılması önerisiyle OYK’ya sunulan dosya sayılarında önemli bir değişim görülmezken, yaptırım önerisiyle OYK’ya sunulan dosya sayısında ve sonuçlandırılmamış dosya sayısında belirgin bir artış olduğu görülmektedir. Sonuçlandırılmamış dosya sayısındaki artış, haksız rekabetle mücadele kapsamında istenilmeyen bir durumdur. Esas olan, haksız rekabet dosyalarının OHRMK’lere ulaştıktan sonra mümkün olan en kısa sürede görüşülüp karara bağlanmasıdır. OHRMK’larca döneminde sonuçlandırılmadan bir Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 232 sonraki döneme bırakılan dosyalar, haksız rekabet davranışında bulunan meslek mensubunun ceza almasını geciktirerek sorunla mücadeleyi olumsuz etkileyecektir. OHRMK’ların yaptırım önerisiyle OYK’lara sundukları dosya sayısı meslekteki haksız rekabet sorununun boyutunu göstermesi açısından önem arz ettiğinden, aşağıda ayrı bir başlık halinde analiz edilmiştir. 5.3. Oda Haksız Rekabetle Mücadele Kurullarının Yaptırım Önerisiyle Oda Yönetim Kurullarına Sundukları Haksız Rekabet Dosya Sayısı 5.3. Oda Haksız Rekabetle Mücadele Kurullarının Yaptırım Önerisiyle Oda Yönetim Kurullarına Sundukları Haksız Rekabet Dosya Sayısı 2008-2022 yılları arasında OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş olduğu haksız rekabet dosya sayısı Grafik 3’te sunulmuştur. Grafik 3: OHRMK’ların Yaptırım Önerisiyle OYK’lara Sundukları Haksız Rekabet Dosya Sayısı Grafik 3: OHRMK’ların Yaptırım Önerisiyle OYK’lara Sundukları Haksız Rekabet Dosya Sayısı OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş olduğu haksız rekabet dosya sayısında 2016 sonrası dönemde belirgin bir artış olduğu görülmektedir. 2016 yılından 2021 yılına kadar sürekli artan dosya sayısı 2022 yılında azalmıştır. 2022 yılındaki bu azalış, aynı yıldaki sonuçlandırılmamış dosya sayısıyla açıklanabilir (Grafik 2). 2022 yılında sonuçlandırılmamış dosya sayısı 2.835 adet ile incelenen tüm yıllar içerisindeki en yüksek sayıya ulaşmıştır. 2016 yılı öncesi dönemde en fazla 880 adete ulaşan yaptırım önerisiyle OYK’ya sunulan dosya sayısı, 2016 yılı sonrası dönemde artarak 1.422 ve 1.906 adetlerle incelenen dönemdeki en yüksek sayıya ulaşmıştır. 2008-2022 dönemi incelendiğinde, OHRMK’ların yaptırım önerisiyle 233 Ünal Arslan / Metin Kılıç Ünal Arslan / Metin Kılıç OYK’lara sunmuş olduğu haksız rekabet dosya sayılarında belirgin bir artış olduğu görülmektedir. Haksız rekabet konusunda uzmanlaşmış kurullar olarak ifade edebileceğimiz OHRMK’ların incelemiş oldukları dosyalarda haksız rekabet davranışını kati olarak tespiti, muhasebe mesleğinde bu sorunun mevcudiyetini kanıtlarken aynı kurulun yaptırım önerisiyle OYK’lara sunmuş olduğu dosya sayılarında yıllar itibarıyla belirgin bir artışın olması, mesleki değerleri ve kuralları hiçe sayan, kişisel çıkarlarını her şeyin önünde gören bir meslek mensubu grubunun varlığını ve bu kişilerin mesleğe, meslek mensuplarına ve tüm topluma karşı olan sorumluluklarını yerine getirmeksizin mesleki faaliyetlerine devam ettiklerini göstermektedir. 5.4. Oda Haksız Rekabetle Mücadele Kurullarının Yaptırım Önerisiyle Oda Yönetim Kurullarına Sunmuş Oldukları Dosyaların Dağılımı OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş oldukları haksız rekabet dosya sayısı, HRRYY’de “Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet” başlığını taşıyan 6. madde, “Ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet” başlığını taşıyan 7. madde ve “Reklam yoluyla haksız rekabet” başlığını taşıyan 8. madde kapsamındaki dosyalardan oluşmaktadır. 2008-2022 yılları arasında OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş oldukları dosyaların madde 6, madde 7 ve madde 8 kapsamında dağılımı Grafik 4’te sunulmuştur. 5.3. Oda Haksız Rekabetle Mücadele Kurullarının Yaptırım Önerisiyle Oda Yönetim Kurullarına Sundukları Haksız Rekabet Dosya Sayısı Grafik 4: OHRMK’ların Yaptırım Önerisiyle OYK’lara Sunmuş Oldukları Dosyaların Dağılımı Grafik 4: OHRMK’ların Yaptırım Önerisiyle OYK’lara Sunmuş Oldukları Dosyaların Dağılımı 234 \| Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş oldukları dosyaların dağılımı incelendiğinde; HRRYY madde 7 ve madde 8 kapsamındaki haksız rekabet dosya sayılarının 2016 yılı sonrasında durağan bir seyir izlediği buna karşın madde 6 kapsamındaki haksız rekabet dosya sayısında belirgin bir artış olduğu görülmektedir. 2022 yılında madde 6 kapsamındaki dosya sayısında görülen azalmayı sonuçlandırılmamış dosya sayısındaki artışla açıklayabiliriz. Şöyle ki; 2022 yılında sonuçlandırılmamış dosya sayısındaki rekor artış yaptırım önerisiyle OYK’lara sunulan dosya sayısını azaltırken (Grafik 3) aynı zamanda madde 6 kapsamındaki dosya sayısını da azaltmıştır. Sonuçlandırılmamış dosya sayısındaki bu artışın etkisi, madde 7 ve madde 8’de de kısmen görülmektedir. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında, madde 7 ve 8 kapsamında toplanan haksız rekabet dosya sayılarında önemli bir değişiklik görülmezken, madde 6 kapsamındaki haksız rekabet dosya sayısında belirgin bir artış olduğu görülmektedir. Aynı durumun 2008-2022 dönemi için de geçerli olduğunu söyleyebiliriz. Ayrıca 2008-2022 döneminde OHRMK’ların OYK’lara yaptırım önerisiyle sunduğu dosyaların dağılımında en fazla dosya sayısının madde 6 kapsamındaki dosyalardan oluştuğu görülürken, ikinci sıranın madde 7, üçüncü sıranın ise madde 8 kapsamındaki dosyalardan oluştuğu görülmektedir. 5.5. Meslek Mensupları Arasında ve İş Sahipleriyle İlişkilerde Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 5.5. Meslek Mensupları Arasında ve İş Sahipleriyle İlişkilerde Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 5.5. Meslek Mensupları Arasında ve İş Sahipleriyle İlişkilerde Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 2008-2022 döneminde, HRRYY madde 6; “Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet” kapsamındaki dosyaların dağılımı Grafik 5’te sunulmuştur. Ünal Arslan / Metin Kılıç \| 235 Grafik 5: Meslek Mensupları Arasında ve İş Sahipleriyle İlişkilerde Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Grafik 5: Meslek Mensupları Arasında ve İş Sahipleriyle İlişkilerde Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet kapsamındaki dosya sayısının dağılımı incelendiğinde; 2016 yılı sonrası dönem dosyaların dağılımında, toplam dosya sayısının %59,1’i 6/a bendinde, %14,8’i 6/f bendinde ve %7,7’si 6/ı bendinde toplanarak ilk üç sırayı oluşturduğu görülmektedir. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında; bu kapsamdaki dosyaların %50,5’inin 6/a bendinde toplandığı, ikinci sıranın %12,3 ile 6/d bendinde, üçüncü sıranın %12,2 ile 6/ı bendinde toplandığı 2016 yılı öncesi dönemde, birinci ve üçüncü sıranın değişmediği görülmektedir. 2008-2022 dönemi incelendiğinde, 6. madde kapsamında en fazla haksız rekabet dosya sayısının 6/a bendinde toplandığını görmekteyiz. Ayrıca 6/a bendi; “Muhasebe ve denetim standartlarına uymamak, mesleki faaliyetlerin yürütülmesinde gereken mesleki özeni göstermemek” HRRYY’de ifade edilen haksız rekabet halleri içinde en fazla ihlal edilen dolayısıyla incelenen 2008-2022 döneminde her yıl en fazla haksız rekabet dosyasının olduğu benttir. 236 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 5.6. Ücret ve Diğer Mali Nitelikteki Uygulamalarla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 5.6. Ücret ve Diğer Mali Nitelikteki Uygulamalarla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 2008-2022 döneminde, HRRYY madde 7; “Ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet” kapsamındaki dosyaların dağılımı Grafik 6’da sunulmuştur. Grafik 6: Ücret ve Diğer Mali Nitelikteki Uygulamalarla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Grafik 6: Ücret ve Diğer Mali Nitelikteki Uygulamalarla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet kapsamındaki dosya sayısının dağılımı incelendiğinde; 2016 yılı sonrası dönem dosyaların dağılımında, toplam dosya sayısının %39,7’si 7/a bendinde, %16,6’sı 7/ğ bendinde ve %9,7’si 7/c bendinde toplanarak ilk üç sırayı oluşturduğu görülmektedir. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında; bu kapsamdaki dosyaların %36,2’sinin 7/a bendinde toplandığı, ikinci sıranın %13,5 ile 7/d bendinde, üçüncü sıranın %11,1 ile 7/f bendinde toplandığı 2016 yılı öncesi dönemde, ilk sıranın değişmediği görülmektedir. 2008-2022 dönemi incelendiğinde, 7. madde kapsamında en fazla haksız rekabet dosya sayısının, “Asgari ücret tarifesinin altında ücret talep etmek veya ücretsiz hizmet vermek” olarak ifade edilen 7/a bendinde toplandığını görmekteyiz. Bu bent aynı zamanda incelenen 2008-2022 döneminde 2008 ve 2011 yılları hariç, her yıl 7. madde kapsamında en fazla haksız rekabet dosyasının toplandığı benttir. Ünal Arslan / Metin Kılıç \| 237 5.7. Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 2008-2022 döneminde, HRRYY madde 8; “Reklam yoluyla haksız rekabet” kapsamındaki dosyaların dağılımı Grafik 7’de sunulmuştur. Grafik 7: Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 5.7. Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 5.7. Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı 2008-2022 döneminde, HRRYY madde 8; “Reklam yoluyla haksı rekabet” kapsamındaki dosyaların dağılımı Grafik 7’de sunulmuştur. 2008-2022 döneminde, HRRYY madde 8; “Reklam yoluyla haksız rekabet” kapsamındaki dosyaların dağılımı Grafik 7’de sunulmuştur. Grafik 7: Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Grafik 7: Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Grafik 7: Reklam Yoluyla Haksız Rekabet Kapsamındaki Dosyaların Dağılımı Reklam yoluyla haksız rekabet kapsamındaki dosya sayısının dağılımı incelendiğinde; 2016 yılı sonrası dönem dosyaların dağılımında, toplam dosya sayısının %33,1’i 8/e bendinde, %15,1’i 8/a bendinde ve %13,1’i 8/f bendinde toplanarak ilk üç sırayı oluşturduğu görülmektedir. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında; aynı sıralamanın %25,5, %17,8 ve %16,4 oranlarıyla korunduğunu görmekteyiz. Bu durum, 2008- 2022 dönemi için de aynı sıralamanın devam etmesini sağlamıştır. 5.8. Haksız Rekabet Dosya Sayılarının SMMM Odalarıyla YMM Odaları Arasında Dağılımı 5.8. Haksız Rekabet Dosya Sayılarının SMMM Odalarıyla YMM Odaları Arasında Dağılımı 2008-2022 dönemi haksız rekabet dosya sayılarının SMMM odalarıyla YMM odaları arasında dağılımı Grafik 8’de sunulmuştur. 38 \| Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi 238 Grafik 8: Haksız Rekabet Dosya Sayılarının SMMM Odalarıyla YMM Odaları Arasında Dağılımı Grafik 8: Haksız Rekabet Dosya Sayılarının SMMM Odalarıyla YMM Odaları Arasında Dağılımı Haksız rekabet dosya sayılarının SMMM odalarıyla YMM odaları arasında dağılımını incelediğimizde; 2016 yılı öncesinde ve sonrasında dolayısıyla incelenen 2008-2022 döneminde, SMMM OYK’larının OHRMK’lara göndermiş olduğu dosya sayısının YMM OYK’larının OHRMK’lara göndermiş olduğu dosya sayısından çok daha fazla olduğu görülmektedir. 2017 yılında YMM OYK’larından OHRMK’lara gönderilen dosya sayısı 13 adet ile incelenen dönemdeki en yüksek seviyeye ulaşmışken aynı yıl SMMM odalarında OHRMK’lara gönderilen dosya sayısı 1.412 adettir. Son yıl verisi olan 2022 yılında ise SMMM odalarındaki haksız rekabet dosya sayısı 4.377 iken YMM odalarındaki haksız rekabet dosya sayısı sadece 2’dir. SMMM ve YMM odalarının üye sayılarının karşılaştırılmasıyla da açıklanamayacak bu fark, meslekteki haksız rekabet sorununun YMM odalarının üyelerinden ziyade SMMM odalarının üyeleri arasında mevcut olduğunu göstermektedir. 6. Sonuç Muhasebe meslek mensupları arasında yaşanan haksız rekabet sorunu mesleğin önemli sorunlarından birini oluşturmaktadır. İvedilikle çözülme kavuşturulması gereken bu sorunun çözüm otoritesi, muhasebe meslek odalarının üst birliği olan TÜRMOB’tur. 3568 sayılı Yasa gereği kurulan TÜRMOB, kuruluş sürecini tamamlamasının hemen ardından meslekteki 239 Ünal Arslan / Metin Kılıç \| haksız rekabet sorunuyla mücadeleye başlamıştır. 03.01.1990 tarihinde yayımlanan “Serbest Muhasebeci Mali Müşavir ve Yeminli Mali Müşavirlerin Çalışma Usul ve Esasları Hakkında Yönetmelik” TÜRMOB’un bu konudaki ilk çalışması olarak ifade edilebilirken, günümüze kadar olan süreçte konuyla ilgili pek çok çalışma gerçekleştirmiştir. Bu çalışmaların içinde özellikle TÜRMOB bünyesinde oluşturulan TÜRHAK’ın yapmış olduğu çalışmalar, haksız rekabetle mücadeleye ayrı bir ivme kazandırmıştır. Haksız rekabet sorununun çözümüne yönelik yapılan tüm bu çalışmaların sonucunda meslekteki bu sorunla ilgili bir durum tespiti yaparak, haksız rekabet dosya sayılarında ve içeriklerinde zaman içinde meydana gelen değişmeleri ve bu değişmelere göre ifade edilebilecek genellemeleri tespit etmeyi hedefleyen araştırmada, bulgulardan ortaya çıkan sonuçlar aşağıda özetlenmiştir: • OYK kararı ile OHRMK’lara gönderilen dosya sayılarının 2016 yılı öncesi dönemdeki durağan seyri 2016 yılı sonrası dönemde yerini belirgin bir artışa bırakmıştır. 2008-2022 dönemi değerlendirildiğinde bu artışın boyutu açıkça görülmektedir. • 2016 yılı sonrası dönemde, OHRMK’larca OYK’lara sunulan dosyaların dağılımında, ceza verilmesine yer olmadığı önerisiyle ve dosyanın işlemden kaldırılması önerisiyle OYK’lara sunulan dosya sayılarında belirgin bir değişme olmazken, sonuçlandırılmamış dosya sayılarında ve yaptırım önerisiyle OYK’ya sunulan dosya sayılarında önemli bir artış olmuştur. Aynı durum, 2008-2022 dönemi için de geçerlidir. • 2016 yılı sonrası dönemde, OHRMK’ların yaptırım önerisiyle OYK’lara sunmuş olduğu dosyaların dağılımında, HRRYY madde 7 ve madde 8 kapsamındaki dosya sayılarında durağan bir seyir izlenirken madde 6 kapsamındaki dosya sayılarında belirgin bir artış olmuştur. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında, madde 7 ve 8 kapsamında toplanan dosya sayılarında önemli bir değişiklik görülmezken, madde 6 kapsamındaki dosya sayısında belirgin bir artış mevcuttur. Aynı durum, 2008-2022 dönemi için de geçerlidir. Ayrıca 2008-2022 döneminde OYK’ya yaptırım önerisiyle sunulan dosyaların dağılımında en fazla dosya sayısı madde 6 kapsamındaki dosyalardan oluşurken, ikinci sırayı madde 7, üçüncü sırayı ise madde 8 kapsamındaki dosyalar oluşturmuştur. • 2016 yılı sonrası dönemde, “Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet” kapsamındaki dosya sayılarının • 2016 yılı sonrası dönemde, “Meslek mensupları arasında ve iş sahipleriyle ilişkilerde haksız rekabet” kapsamındaki dosya sayılarının 240 Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi dağılımında en fazla dosya sayısı sırasıyla 6/a, 6/f, 6/ı bentlerinde toplanmıştır. 6. Sonuç 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında, 2016 yılı öncesi dönemde 6/a, 6/d, 6/ı biçimindeki sıralamayla birinci ve üçüncü sıra değişmemiştir. 2008-2022 döneminde 6. madde kapsamında en fazla haksız rekabet dosya sayısının 6/a bendinde toplandığı tespit edilmiştir. Ayrıca 6/a bendi, HRRYY’de ifade edilen haksız rekabet halleri içinde en fazla ihlal edilen, dolayısıyla incelenen 2008-2022 döneminde her yıl en fazla haksız rekabet dosyasının olduğu benttir. • 2016 yılı sonrası dönemde, “Ücret ve diğer mali nitelikteki uygulamalarla haksız rekabet” kapsamındaki dosya sayılarının dağılımında, en fazla dosya sayısı sırasıyla 7/a, 7/ğ, 7/c bentlerinde toplanmıştır. 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığında, 2016 yılı öncesi dönemde 7/a, 7/d, 7/f biçimindeki sıralamayla birinci sıranın değişmediği görülmüştür. 2008-2022 döneminde 7. madde kapsamında en fazla haksız rekabet dosya sayısının 7/a bendinde toplandığı tespit edilmiştir. • 2016 yılı sonrası dönemde, “Reklam yoluyla haksız rekabet” kapsamındaki toplam dosya sayılarının dağılımında, en fazla dosya sayısı sırasıyla 8/e, 8/a, 8/f bentlerinde toplanmıştır. 2016 yılı öncesi dönemde de aynı sıralama mevcuttur. Bu durum, 2008-2022 dönemi için de aynı sıralamanın devam etmesini sağlamıştır. • Haksız rekabet dosya sayılarının SMMM odalarıyla YMM odaları arasında dağılımında, hem 2016 yılı sonrasında hem de 2016 yılı öncesinde SMMM OYK’larının OHRMK’lara göndermiş olduğu dosya sayısının YMM OYK’larının OHRMK’lara gönderdiği dosya sayısından çok daha fazla olduğu tespit edilmiştir. Araştırmada, haksız rekabet dosyalarının yıllar itibarıyla içerik yönünden bazı değişiklikler gösterse de HRRYY’nin belirli maddelerinde ve bu maddelerin belirli bentlerinde yoğunlaştığı belirlenmiştir. Haksız rekabet dosyalarının yoğun olarak toplandığı haksız rekabet hallerinin bilinmesinin sorunun çözümüne katkı sağlayacağı düşünülmektedir. Şöyle ki, sorunla mücadele kapsamında dosya sayılarının yoğun olarak toplandığı haksız rekabet hallerine öncelik verilmesi ve bu alanlara özel mücadele yöntemleri geliştirilerek mücadele kapsamının genişletilmesiyle bu dosya türlerinin sayısında azalma olacağı düşünülmektedir. 2016 yılı sonrası dönem haksız rekabet dosyalarının analiz edilerek 2016 yılı öncesi dönem sonuçlarıyla karşılaştırıldığı ve böylece 2008-2022 dönemine 241 Ünal Arslan / Metin Kılıç ait değerlendirmenin de yapıldığı araştırmada, muhasebe mesleğinde haksız rekabet sorununun çözümüne yönelik yapılan tüm çalışmalara rağmen sorunun mevcudiyetini koruduğu ve hatta 2016 yılı sonrası dönemde gerek OYK’ların OHRMK’lara gönderdiği dosya sayılarındaki artış gerekse OHRMK’ların yaptırım önerisiyle OYK’lara gönderdiği dosya sayılarındaki artışla sorunun muhasebe mesleğinde kabul edilemeyecek boyuta ulaştığı anlaşılmaktadır. Ancak bu durum sorunun çözümsüz olduğu anlamına da gelmemektedir. Hem TÜRMOB’un hem de TÜRHAK’ın haksız rekabetle mücadele kapsamında bugüne kadar yapmış oldukları çalışmaları bundan sonra da kararlılıkla sürdürmeleri ve gerekiyorsa yeni çözüm önerileri geliştirerek uygulamaları ile meslekteki haksız rekabet sorununun çözümü konusunda istenilen başarıya ulaşılacağı düşünülmektedir. 6. Sonuç 242 \| Muhasebe Mesleğinde Haksız Rekabet Dosya Analizi: 2008-2022 Dönemi KAYNAKÇA Akarsu, A. (2015). Muhasebecilik Mesleğinde Haksız Rekabet Olgusu: Sivas İlinde Bir Araştırma. Cumhuriyet Üniversitesi: Yayımlanmamış Yüksek Lisans Tezi. Arslan, Ü. (2020): Muhasebe Mesleğinde Haksız Rekabetin Boyutu: Türkiye’de Muhasebe Meslek Mensuplarına Yönelik Bir Araştırma, Karabük Üniversi- tesi: Yayımlanmamış Doktora Tezi. Aslan, Ü., ve Özdemir, G. (2020). Muhasebe Meslek Mensuplarının Almış Oldukları Disiplin Cezalarının 2009- 2019 Yılları Arasındaki Durumu Üzerine Bir İnceleme. Muhasebe Bilim Dünyası Dergisi, 22 (Özel Sayı), 162-175. Ekergil, V. (2018). Muhasebe Mesleğinde Haksız Rekabet- Disiplin İlişkisi, Süreci ve Analizi. Eskişehir: Nisan Kitabevi. Karakaya, G., ve Biçer, A., A. (2020). Muhasebe Meslek Mensuplarının Haksız Rekabet Algılarının Analitik Hiyerarşi Süreci Yöntemi ile Değerlendiril- mesi, İşletme Araştırmaları Dergisi, 12 (2), 1149-1158. Ömürbek, V., ve Türkoğlu, T. (2013). Muhasebe Meslek Mensupları Arasında Yaşanan Haksız Rekabet Üzerine Bir Araştırma. Balıkesir University Jour- nal of Social Sciences Institute. 16(30), 115-149. Sak, E. (2013). Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Müşavirlik Mesleğinde Haksız Rekabet Olgusu. Atılım Üniversitesi: Yayımlanmamış Yüksek Lisan Tezi. TÜRMOB (2007), Serbest Muhasebeci Mali Müşavirlik ve Yeminli Mali Mü- şavirlik Mesleklerine İlişkin Haksız Rekabet ve Reklam Yasağı Yönetme- liği. (Erişim Tarihi:19.06.2023), http://www.resmigazete.gov.tr/eski- ler/2007/11/20071121-6.htm TÜRMOB (2008-2022). Faaliyet Raporları. (Erişim tarihi: 25.06.2023) https://www.turmob.org.tr/FaaliyetRaporlari/5e23267b -2c69-46ee-9bf6-0dab32e584b/turmob TÜRMOB (2011), Odalar Arası Uygulama Birlikteliği Haksız Rekabet İşlemleri Disiplin İşlemleri. Ankara: TÜRMOB Yayınları No:422. Bölüm 14 1 T.C. Kütahya Dumlupınar Üniversitesi Lisansüstü Eğitim Enstitüsü Uluslararası Ticaret Ve Finansman Ana Bilim Dalı Yüksek Lisans Öğrencisi , Mail: sahin.d@turkuazmmusavirlik.com Orcid: 0000-0003-1424-2089 2 T.C. Kütahya Dumlupınar Üniversitesi Tavşanlı Uygulamalı Bilimler Fakültesi mehmet.gol@ d d d 2 T.C. Kütahya Dumlupınar Üniversitesi Tavşanlı Uygulamalı Bilimler Fakültesi mehmet.gol@ dpu.edu.tr, Orcid :0000-0001-8411-239X Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik Eğitimi Şahin Daştan1 Mehmet Göl2 1 T.C. Kütahya Dumlupınar Üniversitesi Lisansüstü Eğitim Enstitüsü Uluslararası Ticaret V Finansman Ana Bilim Dalı Yüksek Lisans Öğrencisi , Mail: sahin.d@turkuazmmusavirlik.com Orcid: 0000-0003-1424-2089 Özet Özet Dünyadaki teknoloji alanındaki gelişmelerin akabinde ortaya çıkan dijitalleşmenin etkisi ile pek çok dalda ve sektörde olduğu gibi muhasebe mesleğinde radikal değişimler olmuştur. Muhasebe mesleğini icra edenlerin dijital uygulamalara geçme isteği ve benimsenmesi bütün Dünyayı etkisi altına alan Covid-19 salgını ile sürat kazanmıştır. Dijitalleşme ile meslek mensupları istedikleri mekânda çalışarak zaman kaybını önleyerek mükelleflerini daha nitelikli hizmet verebilirler. Muhasebe eğitimindeki en büyük gaye; kendisini ifade eden, teknik donanıma sahip, dijital çağın gereksinimlere uygun nitelikli muhasebe meslek mensuplarının mesleğe kazandırılmasıdır. Muhasebe meslek mensupları Dünya’da ve ülkemizde yaşanan değişimler karşısında donanımlı olabilmeleri için yalnız okuldan alınan mesleki eğitimleri yeterli görmeyip, muhasebe mesleğini icra ettiği sürece mesleki eğitimini sürdürmesi zorunluluk haline gelmiştir. Zorunlu, sürekli veya yaşam boyu eğitim olarak nitelendirilen eğitim yaklaşımları, muhasebe mesleğini icra edenlerin Dünya’da yaşanan gelişmelere ayak uydurabilmesini gereklilik haline getirmiştir. Muhasebe meslek mensupları tarafından mesleki bilgilerin ve yeteneklerin yaşanılan döneme göre güncellenmesi, muhasebe mesleğini icra ettiği sürece ilgili kurumlar tarafından mesleki eğitim programlarına katıldıkları sürece mümkün olacaktır. Bu çalışmada, muhasebe meslek mensuplarının mesleki yetkinlik ile ilgili problemlerin tanımlanması ve tanımlan sorunların çözülmesinde sürekli eğitimin zorunluluğu ve bu kapsamda düzenlenen https://doi.org/10.58830/ozgur.pub401.c1762 https://doi.org/10.58830/ozgur.pub401.c1762 243 244 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... eğitim programlarının nasıl daha verimli hale getirilebilmesi üzerinde durulmuştur. Çalışma 6 bölümden oluşmaktadır. Çalışmanın ilk bölümü giriş bölümünden oluşmaktadır. 2. bölümde dijital dönüşümün muhasebe departmanına etkisinden muhasebe mesleğini yeniden şekillendirmesinden bahsedilmiştir. Ayrıca dijital dönüşümün muhasebe mesleğini icra edenlere yaşattığı bazı zorluklar bahsedilmiştir. Belirtilen zorluklar şunlardır: Dijital dönüşüm stratejinin olmaması, yönetim ile muhasebe departmanları arasında koordinasyon eksiklikleri, dijital dönüşümde yetkinlik eksiklikleri, dijital dönüşümde siber güvenliğinin dikkate alınmaması, dijital dönüşümün hızlı değişmesidir. 3. bölümde muhasebe çalışanlarının geliştirilebileceği bazı teknolojik ve temel yetkinliklerden bahsedilmiştir. Belirtilen teknolojik yetkinlikler şunlardır: Sosyal Medya, veri analitiği, müşteri ilişkileri yönetimi, video konferans, dijital pazarlama- seo, e-posta ile pazarlama, içerik pazarlaması ve siber güvenliktir. 4. bölümde teknolojik gelişime dönük eğitimin işletmelere sağladığı bazı avantajlardan bahsedilmiştir. Belirtilen avantajlar şunlardır: Para tasarrufu sağlaması, zaman kazandırması, rekabet avantajı teşvik etmesi, güven artırması ve ekip oluşturmaya olanak sağlamasıdır. 5. bölümde teknolojik eğitim programlarının başarılı uygulanması için yapılması gerekenlerden bahsedilmiştir. Yapılması gerekenler şunlardır: Modern eğitim vizyonu geliştirmek, altyapıya yatırım yapmak, öğrenci teknoloji modeline karar vermek, sanal bir öğrenme ortamı yaratmak, eğitime ve mesleki gelişime vurgu yapmak, sunuculara, depolamaya ve yazılıma yapılan yatırımları en aza indirmek, ders kitaplarını değiştirmek, esnek olmak, devlet yardımlarından yararlanmak gibi ipuçlarından bahsedilmiştir. 6.bölüm sonuç bölümünden oluşmaktadır. 2.1 Dijital Dönüşümün Muhasebe Departmanına Etkisi Dünyadaki teknoloji alanındaki gelişmelerin akabinde ortaya çıkan dijitalleşmenin muhasebe mesleğinde yaşattığı etkiler aşağıda belirtilmiştir: (First Intuition. “The future of accounting: how are digital technologies reshaping the industry?”. Erişim: 25.08.2023.) • Dijital Dönüşüm Teknolojileri Muhasebe Meslek Mensuplarına Destekleyecek, Onların Yerini Almayacak Dijitalleşme ile birlikte muhasebe süreçlerinin otomasyon sistemleri ile otomatik hale gelmesi ve daha az zaman almasıyla birlikte, muhasebe meslek mensuplarının mükellefleri ile daha fazla irtibat halinde olmaları ve günlük iş operasyonları söz konusu olduğunda danışmanlık hizmetlerini artırmaktadır (Gulin ve diğerleri 2019). • Dijital Dönüşüm Teknolojileri Muhasebe Meslek Mensuplarına Destekleyecek, Onların Yerini Almayacak • Dijital Dönüşüm Teknolojileri Muhasebe Meslek Mensuplarına Destekleyecek, Onların Yerini Almayacak Dijitalleşme ile birlikte muhasebe süreçlerinin otomasyon sistemleri ile otomatik hale gelmesi ve daha az zaman almasıyla birlikte, muhasebe meslek mensuplarının mükellefleri ile daha fazla irtibat halinde olmaları ve günlük iş operasyonları söz konusu olduğunda danışmanlık hizmetlerini artırmaktadır (Gulin ve diğerleri 2019). • Dijital Dönüşüm Teknolojileri Muhasebe Meslek Mensuplarına Destekleyecek, Onların Yerini Almayacak Dijitalleşme ile birlikte muhasebe süreçlerinin otomasyon sistemleri ile otomatik hale gelmesi ve daha az zaman almasıyla birlikte, muhasebe meslek mensuplarının mükellefleri ile daha fazla irtibat halinde olmaları ve günlük iş operasyonları söz konusu olduğunda danışmanlık hizmetlerini artırmaktadır (Gulin ve diğerleri 2019). Muhasebede kullanılan otomasyon sistemleri, verimliliğin artırılmasına yardımcı olduğu, bilgi girişi gibi zaman alan işlemleri sonlandırdığı ve insandan kaynaklanan hata olasılığını azalttığı için muhasebe meslek mensuplarına daha faydalı olmasını sağlıyor. Bu dijital dönüşümün sağladığı faydalar, çeşitli muhasebe ve finans rollerinde uygulanabilmesi nedeniyle meslek genelinde yaygındır. Otomasyon sistemlerinin bakımı ve desteklenmesi için bilişsel teknoloji uzmanlarına ihtiyaç duyulacaktır. 1 Giriş Dijitalleşme yalnızca teknolojik bir yenilik değil aynı zamanda işletme faaliyetlerini ve süreçlerini yönetme biçiminde farklı bir felsefeyi ifade etmektedir. Dijitalleşmenin yarattığı etki ile yaşadığımız dönemde çevremizde ve dünyada değerli dönüşüm ve gelişmeler yaşanmaktadır (Mancini ve diğerleri, 2017). Dijitalleşmenin etkisiyle meydana gelen teknolojiler, kurallar ve insan kaynakları, insanlara dijital teknolojileri benimseterek onlara zorla kullandıracak ve dolayısıyla iş ve ekonomi hayatında önemli ve geri dönüşü olmayan değişimlere yol açacak şekilde sürekli değişen bir ortamın etkisi altında olduklarından işletmeler için zorluklarıdır (Grosu ve diğerleri, 2023). İşletmelerin Dijital geçiş sürecinde işletmeleri yeni teknolojilerin kullanılması, maliyetlerin minimum seviyeye indirilmesi, daha etkili iyi bir yönetim, maksimum faydalı hizmetler, gelişmiş idari işlevler ve daha az hata gibi çok sayıda avantaj ve işlevsellik sunar (Alshirah ve diğerleri, 2021). Dijitalleşmenin yarattığı etki yaşanan bu gelişmelerle beraber muhasebe mesleği A’dan Z’ye etkilenip klasik muhasebeden dijital muhasebeye evrim 245 Şahin Daştan / Mehmet Göl geçirmektedir. Bu kapsamda muhasebe meslek mensuplarının dijital çağın gereksinimlerine uygun nitelik ve yetkinliklere sahip olmaları gerekmektedir. Bu yetkinliklerin kazanılmasındaki baş aktör muhasebe meslek mensuplarına verilecek nitelikli kaliteli eğitim ile olacaktır. Bu çalışmada, dijitalleşmenin muhasebe departmanına etkisinden, zorluklarından, muhasebe meslek mensuplarının temel ve dijital yetkinliklerinden, dijital yetkinliklerin öneminden, teknolojik gelişiminin işletmelere sağladığı avantajlar, personellere verilen teknolojik eğitimi üzerine durulmuştur. 2 Dijital Dönüşümün Muhasebe Departmanına Etkisi ve Muhasebe 2 Dijital Dönüşümün Muhasebe Departmanına Etkisi ve Muhasebe Meslek Mensuplarının Dijital Dönüşümde Karşılaştıkları Zorluklar Meslek Mensuplarının Dijital Dönüşümde Karşılaştıkları Zorluklar 2.1 Dijital Dönüşümün Muhasebe Departmanına Etkisi • Muhasebe Meslek Mensupları Mükelleflerine Daha Fazla Katkı Sunabilecek Muhasebe meslek mensupları, kurumsal anlamda verimlilik kazanımlarının yanı sıra analiz ve öngörü yetenekleriyle değer yaratabilirler. Tahmin ve sunum yeteneklerin yanı sıra veriye dayalı karar alma süreçlerine de öncülük edebilirler. Böylece doğru teknolojileri Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... 246 benimseyen muhasebe için yeni roller, beceriler ve rekabet avantajı fırsatları ortaya çıkmaktadır. Bununla birlikte, sürekli değişen bu teknolojilere ayak uyduramayan muhasebe meslek mensupları rekabet güçlerini, personellerini ve mükelleflerini kaybetmeleri ihtimali de ortaya çıkmıştır. • Muhasebe Meslek Mensupları Tarafından Hazırlanan Verilerin ve Raporların Mükelleflere Görsel Sunum Yapılması Veri görselleştirme oldukça etkileyici bir alandır. Muhasebeciler bu tür uygulamalar yardımıyla ellerindeki verileri ve sonuçları yeni ve yaratıcı yöntemlerle sunma fırsatı bulmaktadır. Böylece sonuçlar daha iyi gösterilebilir, analize yardımcı olunabilir ve veri anlatımına yardımcı olunabilir. Dolayısıyla daha etkili kararlar alınmasını sağlar. Gerek bir mükellef raporu gerekse şirket içi bir analiz olsun, bu araçlar işletmelerin verileri kullanma, değiştirme ve sunma biçimlerini şekillendirebilir. Sonuç olarak daha net, daha ayrıntılı ve daha gelişmiş bulgular elde edilebilir ve bu da rekabet avantajı sağlayabilir. • Muhasebe Meslek Mensupları Tarafından Dijital Teknolojilerin Anlaşılması Dijital teknolojilerden yararlanan muhasebe meslek mensupları için en büyük engellerden biri nereden başlayacaklarına karar vermektir. Muhasebe meslek mensuplarının çabalarını nereye odaklayacaklarını bilmelerini zorlaştıran çok sayıda farklı dijital teknolojiler vardır. Üstelik daha önce dijital teknolojileri kullanmak zorunda kalmamış ve bu teknolojileri anlamayan muhasebe meslek mensuplarının direnci, benimsemeyi daha da engelleyebilir. Bununla birlikte, rekabetçi kalabilmek için benimsemek giderek daha gerekli hale geliyor. Bundan en fazla fayda sağlayacak olan muhasebe meslek mensupları, hangi teknolojinin kendileri için en iyi sonucu vereceğine karar vermeden önce neyi başarmaya çalıştıklarını anlayan muhasebe meslek mensuplarıdır. Çözülmesi gereken sorunun ne olduğunu, bunu çözmek için hangi becerilere ihtiyaç duyulduğunu ve dijital teknolojilerin onlara nasıl yardımcı olabileceğini anlamak, en uygun ve etkili benimsemeyi kolaylaştıracaktır. • Muhasebe Meslek Mensuplarının Teknolojik Yetkinliklerini Geliştirmesi • Muhasebe Meslek Mensuplarının Teknolojik Yetkinliklerini Geliştirmesi Dijital teknolojiler, bir muhasebe meslek mensubunun geliştirebileceği yetkinlikleri genişletmiştir. Veri analitiği, otomasyon ve kodlama gibi dijital teknolojilerdeki beceriler, muhasebe meslek mensuplarının daha etkin, verimli ve güçlü bir şekilde çalışmasına yardımcı olabilir. Ancak, muhasebe meslek mensuplarının tüm bu yetkinlikleri geliştirmeleri zorunlu olmayıp, mükellefler bu yetkinliklere sahip diğer kişilerle işbirliği yapabilirler. Önemli olan, bu yeni olasılıklara karşı bir anlayışa ve açıklığa sahip olmaktır. Muhasebe meslek mensuplarının sorunları çözmek için gereken doğru soruları nasıl soracaklarını bilmeleri gerekir. Sorunu ve dijital teknolojilerin onlara nasıl yardımcı olabileceğini anlamaları gerekir. • Muhasebe Meslek Mensupları Ofislerinde Çalıştırdıkları Personelleri İşe alma ve Personellerini Elde Tutma Metotlarının Değişmesi Teknoloji tutkunu genç kuşaklar çalışma hayatına katıldıkça, beraberinde yeni çalışma yöntemleri de getirmekte ve dolayısıyla işverenler nesiller arası bir ayrımla karşı karşıya kalabilmektedir. Sınırlı dijital yetkinliğe sahip olmayan meslek mensupları ofislerinde çalıştıracak genç personeli cezbetmekte zorlanabilir. Dijital yetkinliğe sahip olan meslek mensupları ofislerinde dijital yetkinliğe sahip 247 Şahin Daştan / Mehmet Göl \| 247 olmayan daha yaşlı ve deneyimli muhasebecileri istihdam etmekte zorlanabilir. Muhasebe mesleğinde yaşanan dijital dönüşüm yeniliklere açık bir zihniyetin önemini bir kez daha vurgulamaktadır. İşverenlerin giderek daha fazla aradığı dijital yetkinlikler, genç kuşakları muhasebe alanına çekebilir. 2.2 Muhasebe Meslek Mensuplarının Dijital Dönüşümde Karşılaştıkları Zorluklar Muhasebe mesleğinde muhasebe meslek mensuplarının dijital dönüşümde karşılaştıkları zorluklar aşağıda belirtilmiştir: (Alhajjar,2023:94-95 ve ERP Cloud Blog,2022 ) • Yönetim İle Muhasebe Departmanları Arasında Koordinasyon Eksikliği Etkili dijital dönüşüm stratejilerinin oluşumuna hem muhasebe departmanından hem de yönetim ekiplerinden katılım sağlanmalıdır. Muhasebe departmanından sorumlu yöneticiler, stratejinin personel tecrübelerini iyileştireceğinden emin olmalıdır. Aynı zamanda yönetim ekibi, dijital dönüşüm projesinin kârlılığı nasıl etkileyeceğine dair tahminler isteyecektir. Muhasebe departmanı ve yönetiminde yeni dijital deneyimin potansiyel faydalarını anlamasına yardımcı olmak için, mevcut ve etkisiz işlemleri önerilen değişikliklerle karşılaştıran bir açıklama yapmaları gerekir. Yapılan açıklamada tahmini tasarrufları vurgulanması gerekir. Maliyetlerdeki azalmayı belirlemek mümkün değilse bile, güncel durumun nasıl tam şeffaflık sağlamadığını, dolayısıyla düzgün bütçe yapmayı ve kayıpları tespit etmeyi imkânsız hale getirdiğini açıklamak gerekecektir(ERP Cloud Blog,2022). • Dijital Yetkinliklerdeki Eksikler Bunların yanı sıra, tüm çalışanlar yeni dijital sistemi anlamayabilir ve süreçlerinde değişiklik yapma konusunda isteksiz olabilirler. Eğitici programların, ikramiye veya ücretli izin gibi teşviklerle birlikte tasarlanması, yeni sürece karşı direnci azaltmaya yardımcı olabilir. Bunlara ilaveten, muhasebe departmanının dinamik ve erişilebilir bir destek ekibine sahip teknoloji tedarikçilerini tercih etmesi, sorunların ortaya çıkması halinde muhasebe departmanının ilgilenildiğini hissetmesine yardımcı olabilir(ERP Cloud Blog,2022). • Dijitalleşme Sürecinde Yaşanan Siber Güvenlik Problemlerini Dikkate Almamak • Etkili Dijital Dönüşüm Stratejilerinin Olmaması • Etkili Dijital Dönüşüm Stratejilerinin Olmaması • Etkili Dijital Dönüşüm Stratejilerinin Olmaması Bir stratejiye sahip olmamak, bir dijital dönüşüm girişiminin en önemli sorunudur. Sadece yapay zekâya veya gelişmiş analitiğe yatırım yapmaya karar vermek yeterli değildir. Temel performans göstergelerinin tanımlanmasını, eğitimi, uygulamayı ve bakımı içeren uçtan uca bir plana sahip olmak çok önemlidir. Dijital dönüşüm stratejisinin bir kurumun temel iş stratejisini de desteklemesi gerekir. Bu sadece katılım ve finansman olasılığını artırmaya yardımcı olmakla kalmaz, aynı zamanda genel iş stratejisini departman hedefleriyle birleştirmek muhasebeyi önemli bir paydaş haline getirir(ERP Cloud Blog,2022). 248 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... • Dijitalleşme Sürecinde Yaşanan Siber Güvenlik Problemlerini Dikkate Almamak Muhasebede teknolojinin kullanımının artmasıyla birlikte finansal bilgilerin ve hassas verilerin güvenliği konusunda zorluklar ortaya çıkmaktadır. Muhasebeciler güvenlik konusunda derin bir anlayışa sahip olmalıdır (Alhajjar,2023:94-95). Dijitalleşmeyle birlikte başta siber saldırılar olmak üzere yeni tehditler ortaya çıkmaktadır. Herhangi bir dijital dönüşüm stratejisi, başarı için siber güvenliği hesaba katmalıdır. Ekip, yeni teknoloji seçerken siber güvenlik standartlarını gözden geçirmek ve bir kuruluşun nelerden sorumlu olacağını belirlemek için bilişsel teknolojiler departmanlarına ve potansiyel tedarikçilere danışmalıdır. Bilişsel teknolojiler ekibini karar verme sürecinde döngüde tutmak, kapsamlı bir güvenlik stratejisi sağlar (ERP Cloud Blog,2022). Şahin Daştan / Mehmet Göl \| 249 249 Şahin Daştan / Mehmet Göl \| 249 • Dijital Dönüşümde Sürekli Değişimin Olması Dijital dönüşümün hızla gelişmesi, muhasebe meslek mensuplarınım ileri muhasebe sistemleri ve yeni teknolojilerle ilgilenmesini gerektirmektedir. Örneğin Büyük Veri analizi, güçlü analitik yetenekler ve karmaşık finansal verilerin derinlemesine anlaşılmasını gerektirir (Alhajjar,2023:94-95). • Dijital Dönüşümde Sürekli Değişimin Olması Dijital dönüşümün hızla gelişmesi, muhasebe meslek mensuplarınım ileri muhasebe sistemleri ve yeni teknolojilerle ilgilenmesini gerektirmektedir. Örneğin Büyük Veri analizi, güçlü analitik yetenekler ve karmaşık finansal verilerin derinlemesine anlaşılmasını gerektirir (Alhajjar,2023:94-95). Bir kerede tüm departmanı değiştirmeye çalışmak, felaket sonuçlara yol açabilir. Çok hızlı hareket etmek personeller üzerinde çok fazla baskı oluşturabilir, bütçeyi zorlayabilir ve önemli adımların atlanmasına neden olabilir. Başlangıçta adım adım ilerlemek muhasebe departmanları için özellikle önemlidir. Muhasebe departmanları farklı departmanlardan ve şirket dışından kişilerle muhatap olduğundan ve hassas verilerle çalıştığından, dijital dönüşüm projesi dikkatlice incelenmeli, test edilmeli ve optimize edilmelidir. İdeal olarak, muhasebe departmanları ilk olarak en yoğun emek gerektiren prosedür süreçleriyle başlamalıdır. Bu, düzenli iş akışını engellemeden gelecekteki değişiklikler için zaman kazandıracaktır(ERP Cloud Blog,2022). • Yasal ve düzenleyici dönüşümler Muhasebe ve finansal raporlama yasaları sürekli değişim göstermekte- dir. Bu nedenle Muhasebe meslek mensuplarının yeni yasal ve düzen- leyici mevzuatlara uyum sağlaması gerekmektedir. Örneğin, Uluslara- rası Muhasebe Standartlarının (IFRS) uygulanması ve sürekli değişen mali ve vergi mevzuatına uyum, muhasebe meslek mensupları için zorluk teşkil etmektedir(Alhajjar,2023:94-95). • Yetersiz Destek Bir diğer potensiyel sorun da erişim desteğinden yoksun olmak olabilir. Dijital teknolojiyi hayata geçirmek bir takım zorlukları beraberinde getirebilir ve bir işletmenin BT ekibi her şeyi kendi başına çözemez. Dijital teknoloji sağlayıcılarının erişilebilir olması ve destek hizmetlerini düzenli olarak sunması (eğitimden yardım masasına kadar) müşteri ve çalışan deneyiminde büyük bir fark yaratabilir. Aktif bir destek ekibi, işletmeniz ile tedarikçiniz arasında güven oluşturur ve muhasebe ekiplerinin yeni süreçlere daha iyi uyum sağlamasına yardımcı olur(ERP Cloud Blog,2022). • İşi Kaybetme Korkusu • İşi Kaybetme Korkusu Muhasebe meslek mensupları muhasebe işi düzeyindeki bu niteliksel değişimlerin bir neticesi olarak, muhasebe programlarına ek olarak çıkan yardımcı programlarının bolluğu ve birçok muhasebe aşamanın azalmasından dolayı, işlerini kaybetme tehlikesi yaşamaktadır. (Alhajjar,2023:94-95). • Kültürel Dönüşüm Ve Eğitim ş ğ Dijital dönüşüm aynı zamanda muhasebe meslek mensupları için yeni teknolojiyi ve teknolojiyi kullanabilmenin sürdürülebilir yollarını Dijital dönüşüm aynı zamanda muhasebe meslek mensupları için yeni teknolojiyi ve teknolojiyi kullanabilmenin sürdürülebilir yollarını 250 Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... benimsemeleri ve öğrenmeleri gereken kültürel bir değişimdir. Dijital dönüşüm, muhasebe meslek mensuplarının yetkinliklerini güncel tutmak ve teknolojik gelişmelere ayak uydurmak için sürekli eğitim ve öğrenmeyi gerektirir (Alhajjar,2023:94-95). • Mesleki Ve Etik Zorluklar Bu değişim muhasebeci meslek mensuplarını güvenlik, güvenilirlik ve gizlilik gibi yeni etik zorluklarla karşı karşıya bırakmaktadır. Muhasebe meslek mensupları bu zorluklarla etik ve sorumlu bir şekilde başa çıkabilmeli ve muhasebenin mesleki standartlara ve etik kurallara uygun şekilde hareket etmelidir. (Alhajjar,2023:94-95). • Mesleki Ve Etik Zorluklar Bu değişim muhasebeci meslek mensuplarını güvenlik, güvenilirlik ve gizlilik gibi yeni etik zorluklarla karşı karşıya bırakmaktadır. Muhasebe meslek mensupları bu zorluklarla etik ve sorumlu bir şekilde başa çıkabilmeli ve muhasebenin mesleki standartlara ve etik kurallara uygun şekilde hareket etmelidir. (Alhajjar,2023:94-95). • Verilere Ulaşmanın Zorluğu Muhasebe meslek mensupları dijitalleşme ve kişisel verileri koruma yasalarından dolayı istedikleri verilere ulaşmakta zorluk yaşamaktadır. (Alhajjar,2023:94-95). 3. Muhasebe Mensuplarının Dijital Yetkinliklerin geliştirilmesi 3. Muhasebe Mensuplarının Dijital Yetkinliklerin geliştirilmesi 3.1 Dijital Yetkinlik Dijital anlamda yetkin olmak , dijital süreçlerde ve araçlarda etkin olmak için yeterli beceri ve kapasiteye sahip olunmasını ifade etmektedir. (Aksu ve Dalgılıç, 2019: 56). Dijital yetkinlikte kişinin temel bilişim araçlarına ve bilgisayar programlarına hâkim olması ve bu alanda bilgi, beceri ve tutumunu geliştirmesi ilk adım olarak kabul edilmektedir. Çünkü dijital yetkinlik, teknik bilgiden üretken ve stratejik kişisel yetkinliğe ve bilişsel becerilere uzanan bir süreçtir. Bu nedenle, dijital yetkinlik, bireyin bilgiye erişmek, bilgiyi işlemek ve değerlendirmek için teknolojiyi nasıl kullandığını ve dijital araçlar veya medya ile bilgi üretme ve iletişim kurma becerisini kapsamaktadır (Hatlevik ve Christophersen, 2013: 241).Dijital yetkinlik; yeni dijital teknolojiye yön verecek bilgi ve yeteneklerin kazanılmasıdır. Muhasebe alanında bulut tabanlı muhasebe yazılım uygulamaların yaygın hale gelmesiyle birlikte, bulut tabanlı muhasebe yazılım uygulamanız zaten birçok muhasebe görevini dijital olarak gerçekleştirebilir, ancak dijital becerileriniz burada bitmemelidir. Muhasebe meslek mensupları örneğin, sosyal medya uygulamalarını kullanarak içeriği çevrimiçi ve e-posta yoluyla paylaşarak pazarlama faaliyetlerini bir sonraki seviyeye taşıyabilir. Teknolojinin hem muhasebe meslek mensupların hem de mükelleflerin iş hayatında etkin bir rol oynaması, muhasebe alanında sağlam dijital yetkinliklere sahip olmak giderek daha önemli hale geliyor. (Esther Cooke. “8 digital skills accountants need so their practices can succeed”. Erişim: 27.08.2023. https://www.sage.com/en-gb/blog/digital- skills-accountants-need/#skills) • İşi Kaybetme Korkusu • İşi Kaybetme Korkusu Muhasebe meslek mensupları muhasebe işi düzeyindeki bu niteliksel değişimlerin bir neticesi olarak, muhasebe programlarına ek olarak çıkan yardımcı programlarının bolluğu ve birçok muhasebe aşamanın azalmasından dolayı, işlerini kaybetme tehlikesi yaşamaktadır. (Alhajjar,2023:94-95). İşi Kaybetme Korkusu Muhasebe meslek mensupları muhasebe işi düzeyindeki bu niteliksel değişimlerin bir neticesi olarak, muhasebe programlarına ek olarak çıkan yardımcı programlarının bolluğu ve birçok muhasebe aşamanın azalmasından dolayı, işlerini kaybetme tehlikesi yaşamaktadır. (Alhajjar,2023:94-95). • Yeni Çıkan Mevzuatlarla – İşyerinde Kullanılan Teknolojilerin Çatışması Muhasebe meslek mensuplarının dijitalleşme dönüşümüne hazırlıklı olmamalarının yanı sıra, mevcut muhasebe ve denetim standartlarının yayınlandığı dönemde mevcut bilgi teknolojisinin tam anlamıyla mevcut olmadığı belirtilmektedir. Finansal alanda dijital dönüşüme yeterince önem verilmemiş ve muhasebe modeline henüz yansıtılmamıştır. Genel olarak veri tabanının mevcut olması durumunda muhasebe açıklamalarında önemli bir değişiklik olmayacaktır. (Alhajjar,2023:94-95). • Yeni Çıkan Mevzuatlarla – İşyerinde Kullanılan Teknolojilerin Çatışması • Yeni Çıkan Mevzuatlarla – İşyerinde Kullanılan Teknolojilerin Çatışması • Yeni Çıkan Mevzuatlarla – İşyerinde Kullanılan Teknolojilerin Çatışması Muhasebe meslek mensuplarının dijitalleşme dönüşümüne hazırlıklı olmamalarının yanı sıra, mevcut muhasebe ve denetim standartlarının yayınlandığı dönemde mevcut bilgi teknolojisinin tam anlamıyla mevcut olmadığı belirtilmektedir. Finansal alanda dijital dönüşüme yeterince önem verilmemiş ve muhasebe modeline henüz yansıtılmamıştır. Genel olarak veri tabanının mevcut olması durumunda muhasebe açıklamalarında önemli bir değişiklik olmayacaktır. (Alhajjar,2023:94-95). 251 Şahin Daştan / Mehmet Göl 3. Muhasebe Mensuplarının Dijital Yetkinliklerin geliştirilmesi 3.2 Muhasebe Meslek Mensupları İçin Dijital Yetkinliklerin Önemi İşletmede kullanabilecek tüm araçlardan haberdar olmak, işle ilgili bilgiler elde etmek ve bilinçli kararlar almak için bunlardan yararlanabilmek gerekir. Amaç zamandan ve kaynaklardan tasarruf ederek ve mükelleflere daha kaliteli bir hizmet verebilmektir. Sağlam dijital yetkinliklere sahip olmak, özellikle muhasebe personellerinin Covid-19 pandemisi ile uzaktan çalışmasının giderek yaygınlaştığı günümüzde, muhasebe meslek mensuplarının ve mükelleflerin etkili bir şekilde işbirliği yapmasına ve iletişim kurmasına imkân verir. Muhasebe meslek mensupları tarafından mükelleflerine muhasebe yazılımı, veri analizi araçları ve siber güvenlik dâhil olmak üzere çeşitli dijital yetkinlik konusunda eğitim ve destek verebilir. Muhasebe mensupları mükelleflerine bu hizmetleri vererek mükelleflerin gözünde güvenilir bir danışman olduğunu kanıtlayarak onlarla daha güçlü, daha işbirlikçi ilişkiler kurarlar. (Esther Cooke. “8 digital skills accountants \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... 252 need so their practices can succeed”. Erişim: 27.08.2023. https://www. sage.com/en-gb/blog/digital-skills-accountants-need/#skills) 3.3 Muhasebe Meslek Mensuplarının Sahip Olması Gereken Temel, Dijital Yetkinlikler Ve Çağdaş Muhasebe Meslek Mensubu Profili 3.3.1. Muhasebe Meslek Mensuplarının Sahip Olması Gereken Temel Yetkinlikler 3.3.1. Muhasebe Meslek Mensuplarının Sahip Olması Gereken Temel Yetkinlikler Dijital dönüşümünün temel becerileri, yetenekleri bir çalışanın belirli bir görevi yerine getirmesi veya sosyal ve iş süreçlerinin dijitalleşmesi bağlamında belirli bir faaliyet sonucunu elde etmesi için gerekli olan yetkinlik olarak yorumlanır. Böylece dijital bir muhasebecinin yeterlilikleri hem mesleki görevleri yerine getirmek hem de bir çalışanın dış dünyayla tam teşekküllü etkileşimi ve günlük görevleri çözmek için gereklidir. Dijital ekonomide bir muhasebe meslek mensubunun aşağıdaki temel becerileri tanımlanabilir ( Makaruk,2021:131-132): • Dijital ortamda iletişim ve işbirliği Bir muhasebecinin dijital ortamda, diğer insanlarla etkileşim içinde hedeflerine ulaşmasını sağlayan çeşitli dijital araçları kullanma becerisini ifade eder. • Belirsizlik karşısında kendini geliştirme Bir muhasebecinin, ortaya çıkan yaşam görevleri için kendisi için eğitim hedefleri belirlemesi, diğer gerekli yeterliliklerin çözümlerini ve geliştirme araçlarını (dijital araçların kullanılması dahil) seçmesi gerekir. • Yaratıcı düşünme. Dijital ekonominin sorunlarını çözmek için yeni fikirler üretmek, standart modellerden soyutlamak: mevcut sorunları çözme yollarını yeniden inşa etmek, yeni optimal algoritmalar geliştirmek için alternatif eylem seçenekleri ortaya koymak. • Bilgi ve veri yönetimi. Muhasebe meslek mensubunun gerekli bilgi ve veri kaynaklarını aramak, dijital araçları kullanarak bilgileri algılamak, analiz etmek, ezberlemek ve iletmek ve ayrıca çeşitli kaynaklardan elde edilen verilerle çalışırken alınan bilgileri sorunları çözmek için etkili bir şekilde kullanmak için algoritmalar kullanmaktır. • Bilgi ve veri yönetimi. • Dijital zekâ Dijital zekâ güncel ve henüz yeni piyasaya çıkan dijital teknolojilerin, niteliklerin, dijital uygulamaların, dijital stratejilerin ve dijital kültürün farkındalığını ve uygulamasını içerir. Muhasebe meslek mensupları sürekli bundan sonraki aynı yazılımların kullanmasını beklemek doğru olmaz. Teknolojiliyle meydana gelen gelişmeleri kavramaya ve onları kullanabilmeye hazır olmalılar. • İş zekâsı İş zekâsı Gelecek zamanda muhasebe meslek mensupları iş hayatında zekâlarını kullanarak mali sorunları çözmede çözümler geliştirmeleri ve stratejik olarak işlerine değer katmaları gerekecek. Gelecek zamana uygun hareket etmek için veri analizi, teknoloji, süreç tasarımları, veri, proje ve risk yönetiminde yeterlilik de istenen yetkinlikler arasında olacak. • Analitik beceriler Geleceği düşünen muhasebe meslek mensupları hem teorik hem pratik veri ve analitik görüşüne sahip olmaları gerekir. Her türlü bilgilerin nasıl derlenip, analiz edilip, geleceğe yönelik mali bilgilerin tahmin analizlerini ve bulgularını gerçekleştirmeyi de içerir. • Bilgi ve veri yönetimi. 253 Şahin Daştan / Mehmet Göl \| 2 • Dijital ortamda eleştirel düşünme. Bilgiyi, güvenilirliğini değerlendirmek, gelen bilgi ve verilere dayanarak mantıksal sonuçlar oluşturmak. Bilgiyi, güvenilirliğini değerlendirmek, gelen bilgi ve verilere dayanarak mantıksal sonuçlar oluşturmak. Muhasebe meslek mensupların gelecekteki dönemlerde sahip olması gereken temel yetkinlikler aşağıda belirtilmiştir: (İsmail Tekbaş.“Gelecekte bir muhasebecinin ihtiyaç duyacağı 6 beceri”. Erişim: 28.08.2023 https:// www.parasut.com/blog/bir-muhasebecinin-bilmesi-gerekenler ) • Stratejik Karar Verme • Stratejik Karar Verme Teknolojinin sürekli gelişim halinde olmasından dolayı muhasebe işlemlerimi dijital sistemler tarafından gerçekleştirilmesiyle oluşan boş vakitlerinde muhasebe meslek mensupları stratejik kararlar vereceklerdir. Günümüzde de iş organizasyonlarda stratejik karar alma aşamasına katılımları git gide artış göstermektedir. Stratejik Karar Verme Teknolojinin sürekli gelişim halinde olmasından dolayı muhasebe işlemlerimi dijital sistemler tarafından gerçekleştirilmesiyle oluşan boş vakitlerinde muhasebe meslek mensupları stratejik kararlar vereceklerdir. Günümüzde de iş organizasyonlarda stratejik karar alma aşamasına katılımları git gide artış göstermektedir. • Yaratıcılık Muhasebe meslek mensupların yaratıcı düşünceye sahip olmaları gerektiği ve sürekli gelişen dijitalleşme sürecinde geri kalmamaları gerektiğini ifade eder. Muhasebe sektöründe yaratıcı olmak, potansiyel neticeleri ortaya çıkarmaları, yeni bağlantılar kurarak ve yeni fikirler üretmeyi içerir. • Etkili İletişim Bilgiler etkin bir biçimde 3. kişilerle paylaşılmadığı zaman etkisizdir. Analitik yetkinlikleri, verilen verileri 3. kişilere iletilemezse amacına uygun davranılamaz. Muhasebe meslek mensupları bu sebeplerden dolayı yaptıkları analizlerini kavraması kolay evraklarla sunabilmek 254 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... için her türlü iletişim yetkinliklerini geliştirmelidir. Teknoloji iş hayatında çok önemli bir değere sahip olsa bile, fakat ticari işlemlerde insan etkileşiminin yerini dolduramaz. 3.3.2 Muhasebe Meslek Mensuplarının Sahip Olması Dijital Yetkinlikler Dijital yetkinlik konusunda Janssen ve diğerleri (2013), uyguladıkları anket çalışmasının neticesinde genel dijital yetkinlik çerçevesini ortaya çıkarmışlardır. Araştırmanın neticesinde açıklanan yetkinlikler şunlardır: • Dijital araçların kullanılmasına yönelik beceriler, • Teknolojiyi günlük hayatta kullanma, bilişsel teknolojileri kulanarak mesleki performansı geliştirmek, • Dijital çalışma ortamlarında iletişim kurma, dijital bilgi yönetimi, kişisel verileri koruma ve güvenlik tedbirlerini alma, • Dijital ortamlarda yasal ve etik konularını gözeterek, teknolojiye karşı pozitif düşünceye sahip olma, • Bilişsel teknolojilerini kullanarak ve hayata entegre etme, • Gereksinin duyulan dijital teknolojiler konusunda bilinçli kararlar alma ve mesleki verimliliği artırmak amacıyla dijital teknolojileri faydalı kullanmaktır Aslitürk ve diğerleri (2016) yaptıkları çalışmada dijital ekonomide sahip olunması gereken dijital ve teknik beceriler başlığı altında bilgi iletişim teknolojileri okuryazarlığı, dijital iletişim; bilgi becerileri başlığı altında dijital bilgi işlem becerileri, veri okuryazarlığı, siber güvenlik ve girişimcilik becerileri başlıkları altında ise dijital girişimcilik yetkinlikleri konuları dikkat çekmektedir. Çark (2020), yaptığı çalışmada dijital çağda bireylerin sahip olması beklenen teknik ve kişisel yetkinlikler aşağıda gösterilmiştir: Şahin Daştan / Mehmet Göl \| 255 Dijital Yetkinlikler 1-)Teknik Yetkinlik Zorunlu • Bilişim teknolojileri bilgi ve becerileri • Veri işleme ve analizi • İstatistiki bilgi • Kurumsal işleyiş ve süreçlere ilişkin anlama • Modern ara yüzlerle etkileşime girme becerisi Gerekli • Bilgi yönetimi • Teknoloji ve örgütlenme hakkında disiplinler arası genel • İmalat faaliyetleri ve süreçleri hakkında uzmanlık bilgisi • IT güvenliğine ve verilerin korunması ile ilgili farkındalık Olabilir • Bilgisayar programlama ve kod yazma • Teknoloji hakkında uzmanlık bilgisi • Ergonomi hakkında farkındalık • Hukuki işlere yönelik bilgi ve anlama 2-) Kişisel Yetkinlik Zorunlu • Öz yönetim ve zaman yönetimi • Değişime ayak uydurma becerisi • Ekip çalışması becerisi Dijital Yetkinlikler 1-)Teknik Yetkinlik Zorunlu • Bilişim teknolojileri bilgi ve becerileri • Veri işleme ve analizi • İstatistiki bilgi • Kurumsal işleyiş ve süreçlere ilişkin anlama • Modern ara yüzlerle etkileşime girme becerisi Gerekli • Bilgi yönetimi • Teknoloji ve örgütlenme hakkında disiplinler arası genel • İmalat faaliyetleri ve süreçleri hakkında uzmanlık bilgisi • IT güvenliğine ve verilerin korunması ile ilgili farkındalık Olabilir • Teknoloji ve örgütlenme hakkında disiplinler arası genel • İmalat faaliyetleri ve süreçleri hakkında uzmanlık bilgisi • IT güvenliğine ve verilerin korunması ile ilgili farkındalık Olabilir • Hukuki işlere yönelik bilgi ve anlama • Sosyal Medya y y Muhasebe meslek mensupları hedeflerine ulaşmak için sosyal medya uygulamalarını etkin bir şekilde kullanabilir. Bunun çeşitli yolları vardır. Sosyal medya uygulamaları sektördeki diğer meslek mensupları ile ağ kurmak ve potansiyel mükellef adayları bağlantı kurulabilir. Sosyal medya uygulamaları ile meslek mensupları kendi görüşlerini, sektörel haberleri ve makalelerini paylaşarak marka bilirliği oluşturabilirler. Personel alımlarını bu uygulamalar üzerinden gerçekleştirebilir. Muhasebe meslek mensupları hedeflerine ulaşmak için sosyal medya uygulamalarını etkin bir şekilde kullanabilir. Bunun çeşitli yolları vardır. Muhasebe meslek mensupları hedeflerine ulaşmak için sosyal medya uygulamalarını etkin bir şekilde kullanabilir. Bunun çeşitli yolları vardır. 2-) Kişisel Yetkinlik Zorunlu • Öz yönetim ve zaman yönetimi • Değişime ayak uydurma becerisi • Ekip çalışması becerisi • Sosyal beceriler • İletişim becerileri • İletişim becerileri Gerekli • Yeni teknolojilere güvenme • Sürekli gelişim ve yaşam boyu öğrenmeye yatkınlık • Sürekli gelişim ve yaşam boyu öğrenmeye yatkınlık 256 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... Dijital çağın çalışma koşullarına ve beklentilerine uyum sağlanmasını kolaylaştıran bu yetkinliklerin ve dijitalleşmenin neden tercih edilmesi gerektiğine ilişkin yapılan çalışmalarda ise aşağıdaki nedenler sıralanmaktadır (PwC, 2017): • Çalışanların yapılan işlerde dijital çözümleri kullanması, verimlilik artışı göstermektedir. • Dijitalleşme yapılan işlerde hız kazandırmakla birlikte, rekabet gücü artışı göstermektedir. Ayrıca yapılan işin vaktinden önce bitirilmesi, maliyetlerin minimuma indirilmesi ve elde edilen faydaya bağlı olarak kalite beklentilerine uygunluk ve müşteri memnuniyetini artırmaktadır. • Dijitalleşme finansal performansa direkt katkı sağlamaktadır. • Yönetimin karar alıp vermede süreci kısalmaktadır. • İnovasyon ve ticarileşme aşamaları surat göstermektedir. • Yeni faaliyet alanları veya işlerin yeni versiyonları oluşmaktadır. • Toplumsal ve ekonomik alanlarında hızla gelişim göstermektedir. Muhasebe meslek mensupların dijital çağa ayak uydurabilmeleri için sahip olması gereken dijital yetkinlikler aşağıda belirtilmiştir: Esther Cooke. Muhasebe meslek mensupların dijital çağa ayak uydurabilmeleri için sahip olması gereken dijital yetkinlikler aşağıda belirtilmiştir: Esther Cooke. “8 digital skills accountants need so their practices can succeed”. Erişim: 27.08.2023. https://www.sage.com/en-gb/blog/digital-skills-accountants- need/#skills) “8 digital skills accountants need so their practices can succeed”. Erişim: 27.08.2023. https://www.sage.com/en-gb/blog/digital-skills-accountants- need/#skills) • Video Konferans Muhasebe meslek mensuplarının video konferans yazılım programları vasıtasıyla, mükelleflerle bir araya gelip yüz yüze toplantılara gerek kalmadan onlarla mekân fark etmeksizin uzaktan yüz yüze toplantılar yapmalarına olanak tanır. Video konferansı yöntemi ile zamandan tasarruf edebilir ve seyahat masraflarını azaltabilir, ayrıca mükellefler arasındaki işbirliğini ve iletişimi geliştirebilir. Video konferansı kullanarak, farklı ofislerde, farklı şehirlerde, farklı ülkelerde bulunan mükelleflerle kolayca bağlantı kurabilir, birlikte daha verimli çalışmaya olanak tanır, mükelleflere zamanında ve doğru bilgi verilir. • Dijital Pazarlama ve Seo Muhasebe meslek mensuplarının dijital pazarlama vasıtasıyla çevrimiçi görünürlüğünü artırması, daha geniş çevrelere ulaşmasına ve potansiyel mükellefler bulabilir. Muhasebe meslek mensuplarının oluşturduğu web sitesinin arama motoru sonuçlarındaki sıralamasını iyileştirmek için arama motoru optimizasyonunu (SEO) ve potansiyel mükellefi çekmek için tıklama başına ödeme reklamcılığı gibi diğer dijital pazarlama stratejilerini kullanabilir. • Veri Analitiği Genellikle muhasebe yazılımınızla entegre olan güçlü analiz araçları uygulamaları mevcuttur. Bu analiz araçları ile finansal verilerindeki eğilimleri, kalıpları belirlemek ve daha doğru finansal raporlama sağlamak için kullanabilir. Veri analitiği ile mükelleflerin satışlarında belirli bir faaliyet döneminde yaşanan azalmayı tespit etmenize ve bunu bir raporlama hatası veya gelecekte ele alınabilecek sezonluk bir eğilim olarak tanımanıza yardımcı olabilir. Ayrıca veri analitiği 257 Şahin Daştan / Mehmet Göl ile temel performans göstergelerini analiz etmek için kullanabilir ve şirketin kârlılığını artırmak için maliyetlerin azaltılabileceği alanları belirlemek gibi veriye dayalı öngörüleri kullanarak mükelleflerin karar verme süreçleri düzeltilebilir. • Müşteri İlişkileri Yönetimi Müşteri ilişkileri yönetimi (CRM) yazılımlarını kullanarak mükelleflerle ilgili etkileşimlerini daha iyi yönetilmesine ve analiz edilmesi olanak tanır, böylece mükellef davranışı ve tercihlerine ilişkin kişiselleştirilmiş destek ve öngörüler sağlanabilir. Mükellef verilerini analiz ederek yeni iş fırsatlarını belirlemek ve mükelleflerin gereksinimlerini ve tercihlerini daha iyi anlayarak gelir tahminlerini geliştirmek için de bir CRM kullanabilir. Aynı zamanda mükellefi elde tutmayı geliştirmek için de değerli bir araçtır. Bunun sebebi, bir süredir hizmetlerinizle etkileşime girmeyen mükellefe hedefli iletişimler ve kişiselleştirilmiş destek sunabilmeniz ve aynı zamanda müşteri etkileşimlerini yönetmek için merkezi bir platform kullanarak ekip üyelerinin aynı hedefler doğrultusunda çalışmasını sağlayabilmenizdir. • Siber Güvenlik Muhasebe meslek mensuplarının siber güvenlik becerileri, mükelleflerinin hassas mali ve finansal verilerini siber tehditlerden korumasına yardımcı olacaktır. Muhasebe meslek mensuplarının muhasebe yazılım uygulamaları, mükellefler adına kişisel, mali ve finansal bilgiler de dâhil olmak üzere birçok gizli veriyi işler. Bu veriler yanlış ellere geçerse ciddi mali kayıplara, itibar kaybına ve hukuki sonuçlara yol açabilir. Muhasebe meslek mensuplarının iyi siber güvenlik becerilerine sahip olarak mükelleflerin verilerini bilgisayar korsanlarından, virüslerden ve diğer siber tehditlerden korumak için uygun güvenlik önlemlerini uygulayabilirler . • İçerik Pazarlaması Muhasebe meslek mensuplarının kendilerini muhasebe sektöründe bir düşünce lideri olarak konumlandırmak için içerik pazarlamayı kullanabilirler. İçerik pazarlaması, muhasebe meslek mensuplarının hedef kitlelerinin gereksinimlerine ve şüphelerine hitap eden blog yazıları, makaleler ve videolar gibi değerli içeriklerin çevrimiçi olarak oluşturulmasını ve paylaşılmasını içerir. Bu şekilde muhasebe meslek mensuplarınca potansiyel mükelleflerin güvenini kazanırlar ve kendilerini, uzmanlık alanlarında uzman olarak konumlandırabilirler. İçerik pazarlamasını kullanmanın muhasebe meslek mensuplarını başka bir faydası da, arama motoru sıralamalarını iyileştirip ve web sitelerine potansiyel mükellefleri çekmesine yardımcı olur. Arama motorları için optimize edilmiş, yüksek kaliteli ve bilgilendirici içerik oluşturarak, potansiyel mükelleflerin çözüm öneriler aradıkları sırada web sitelerinin arama motoru sonuçlarında görünme olasılığını artırabilirler. • E-postayla pazarlama Muhasebe meslek mensuplarının e-posta pazarlaması ile mükelleflerle bağlantı halinde olmanın, onları önemli mali, vergisel ve finansal 258 Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... haberler hakkında bilgilendirmenin ve kişiselleştirilmiş destek sağlamanın uygun maliyetli bir yoludur. Muhasebe meslek mensupları mükelleflerle ekonomik ve mali açıdan görüşlerini paylaşmak, sürekli değişim gösteren vergi kanunu değişiklikleriyle ilgili güncellemeler sağlamak için e-posta bültenlerini kullanabilirler. E-posta pazarlamasını kullanmanın bir diğer yararı da yeni potansiyel mükellefler yaratmasına ve mükellef tabanını büyütmesine yardımcı olabilmesidir. 3.3.3 Çağdaş Muhasebe Meslek Mensubu Profili Muhasebe meslek mensuplarının dijitalleşmenin yarattığı sahip olması gereken profil aşağıda belirtilmiştir (Cenar ve diğerleri,2021:53-54): 259 Şahin Daştan / Mehmet Göl • Verileri analiz etme ve görselleştirme yeteneği, Yapay Zekâ, Bulut Bilişim, Blockchain, Büyük Veri gibi modern araçları yönetme becerisi, • Dijitalleşme ile ortaya çıkan yeni standartlar, dijital uygulamalar ve dijital teknolojilerle ilgili klasik ve yeni beceriler, dijital ve sosyal medyanın oluşturulmasıyla modellenen analitik beceriler, teknolojik beceriler, iş otomasyonu becerileri, iletişim becerileri, • Dijital teknoloji becerilerinin yönetimi, • Dijital teknoloji becerilerinin yönetimi, • Teknik ve analitik beceriler, muhasebe alanına uygulanabilir Bilişsel Teknoloji araçlarının etkinliğini değerlendirme yeteneği, • Gelişmiş programlama dilleri ve hesaplamalara, artırılmış muhasebe uygulamalarına dayalı çevrimiçi iş ortamlarını yönetmeye yönelik özel beceriler, • Veri analizi görevleri için beceriler, iletişim becerileri, veri modelleme becerileri, istatistik ve programlama bilgisi, insan bilgisayar etkileşimi becerileri, • Bilgi teknolojisinde iş analizi beceri ve becerileri, veri analizi ve programlama bilgisi, korelasyonları tespit edecek analitik beceriler, istatistik ve genel programlama dilleri bilgisi, iletişim becerileri, ERP, CRM, Excel, SQL, Power Point gibi yazılımları kullanma becerisi, • Yapay zekâ ve makine öğrenimi becerileri, • Yapay zekâ ve makine öğrenimi becerileri, • Yeni dijital trendleri keşfetme becerileri, yenilikçi Bilişsel Teknolojileri uygulamaları ve hizmetlerini uygulama becerileri, iş modelleri alanında yenilik yapma ve değişimi teşvik etme becerisi, • Dijital beceriler, iletişim becerileri, takım çalışması becerileri, yönetim becerileri, eleştirel düşünme becerileri, veri analizi becerileri, • İş analizi becerileri, bilgi teknolojisi becerileri, programlama dili becerileri, iş iletişimi becerileri, istatistiksel bilgi, modern teknoloji yönetimi ve veri analizi becerileri, • Yapay zekâyı programlayabilmeyi sağlayan teknik beceriler, veri analizi, ileri düzey dijital beceriler, • Genel bilgi teknolojisi bilgisi, bilgi teknolojisi kontrol becerileri bilgisi, işyerinde bilgi teknolojisini kullanma becerisi, yaratıcı beceriler ve hayal gücünün, iletişim becerilerinin, eleştirel düşünmenin geliştirilmesi, • Teknoloji ile ilgili beceriler, alanın dijital yolculuğunu destekleyen teknik beceriler, multidisipliner beceriler, RPA (Robotik Süreç 260 Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... Otomasyonu) ile ilgili beceriler, veri yönetimi, veri programlama bilgisi, istatistik bilgisi gibi beceriler 5. Teknolojik Eğitim Programlarının Başarılı Uygulanması İçin İpuçları Teknoloji gelişmeye devam ettikçe işletmelerin yeni teknolojilerini işletmelerinde nasıl uygulayacaklarını öğrenmeleri gerekiyor. Teknolojinin işletmelerde uygulanması, özellikle personellerin teknolojide yaşanan değişime karşı dirençli olduğu ve teknolojiyi kabullenmediği durumlar korkutucu görünebilir. (E.Flanagan. “How to convince employees to adopt new technology”. Erişim: 01.09.2023. https://www.unboxedtechnology. com/blog/how-to-implement-new-technology-in-the-workplace/). 4. Teknolojik Gelişime Dönük Eğitimin İşletmelere Sağladığı Avantajlar 4. Teknolojik Gelişime Dönük Eğitimin İşletmelere Sağladığı Avantajlar Dijital Teknolojiler yaşadığımız dünyada hayatımız vazgeçilemez bir parçası haline gelmiştir. Güncel yaşantımızda cep telefonları, akıllı TV’ler, bilgisayarlar, sesli mesajlaşma, yazılım uygulamaları vb dijital teknolojileri kullanıyoruz. İşletme personelleri bu teknolojileri iş hayatında verimli şekilde kullanacaklarını bilmiyorsa üretkenliğini ve iş hayatını sekteye uğratabilir. İşletme personellerini teknolojik gelişime dönük eğitimin işletmeye sağladığı bazı avantajlar aşağıda belirtilmiştir: (Protech Service Group. “5 Benefits of Technology Training”. Erişim: 28.08.2023. https://www.psgi.net/5- benefits-of-technology-training/ ve London School of Business & Finance. “How Will Technology Change Accounting in the Future?” Erişim: 28.08.2023. https://www.lsbf.org.uk/blog/online-learning/how-will- technology-change-accounting-in-the-future ): • Para tasarrufu sağlar: Eğitimini başarıyla tamamlayan işletme personelleri sorun gidermeye daha az, verimli ve gelir getiren görevlere daha fazla vakit harcarlar. • Zaman kazandırır: İşletme personelleri kullandıkları uygulamalar hakkında tam donanıma sahip olduklarında daha süratli çalışabilir ve projeleri daha hızlı tamamlayabilirler. • Rekabet avantajı sağlar: Tüm çözümlerini etkin bir şekilde kullanabilen işletmeler, temelleri anlamakta zorlanan işletmelerden daha iyi performans gösterecektir. Zamandan ve paradan tasarruf eden işletmeler bu tasarrufları yenilikçi projelere tahsis edebilirler. • Özgüveni artırır: Kendine güvenen personeller yaptığı işte daha iyi performans gösterenlerdir. Personeller yaptıkları görevi nasıl başarılı bir şekilde yapacaklarını bildiklerinde, pozisyonlarından memnun olma olasılıkları daha yüksek olacaktır. Bu da işletme için daha az iş devri ile sonuçlanır. • Ekip oluşturmaya olanak sağlar: Eğitim, işletme bünyesindeki farklı departmanlardaki personellerim birlikte öğrenmesini ve çalışmasını gerektirir. Daha tecrübeli kullanıcılar devreye girip tecrübesi olmayan kullanıcılara yardım edebilir ve personellerde diğer departmanların nasıl çalıştığına dair bir fikir edinebilmelidir. 261 Şahin Daştan / Mehmet Göl • Üretkenliği artırır: Teknoloji, muhasebe süreçlerinin daha verimli hale getirilmesi için kullanılır; bu, iş akışının daha sorunsuz çalışması ve işin daha verimli bir şekilde tamamlanması anlamına gelir. • Yapılan işlerin doğru olmasını olanak sağlar: Veriler üzerinde çalışmak için yapay zekânın kullanılması insan hatası sorununu ortadan kaldırır ve doğru kullanıldığında soruna ilişkin daha doğru bilgiler verebilir. • Yapılan işlerin güvenli olmasını olanak tanır: Muhasebe firmaları genellikle çok sayıda yüksek riskli bilgi tutar ve verileri çok az riskle saklamanıza ve korumanıza olanak tanıyan birçok gelişmiş program vardır. Eğitim, işletmede teknoloji çözümlerinin ve planlamasının gerekli bir yönüdür. Teknolojiye giderek daha fazla bağımlı hale gelirken, personeller işletmenin yatırım yaptığı yeni teknoloji uygulamalarını nasıl kullanacaklarını bilmeleri önemlidir. 5.1 Teknolojiyi İşletmelerde Kullanılmasının Zorlukları Yeni teknolojiyi işletmelerde kullanılmaya başlamanın bazı zorlukları aşağıda belirtilmiştir: (E.Flanagan. “How to convince employees to adopt new technology”. Erişim: 01.09.2023. https://www.unboxedtechnology. com/blog/how-to-implement-new-technology-in-the-workplace/). Değişime Direnç: ABD’de faaliyet gösteren Robert Half adlı insan kaynakları danışmanlık firması tarafından yapılan bir anket de, işletme yöneticilerinin teknolojide ki yaşanan değişimlere karşı direnci başarılı teknoloji uygulamasının önündeki en büyük engel olarak gördüklerini ortaya çıkardı. İşletme personellerinin direnişi genellikle yeni teknolojileri kullanmayı bilmediklerinden duyulan korkudan veya yeni teknolojiyi kullanma yetkinliklerine sahip olmadıkları için güven eksikliğinden kaynaklanmaktadır. 262 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... Sınırlı Eğitim: İşletmelerin yeni çıkan teknolojileri işletmelerinde kullanmaya başlarken yazılım ve sistem eğitimi çok önemlidir. İşletmeler, personellerine yeni teknolojileri zorla dayatmak yerine, onlara bu yeni çıkan teknolojileri başarıyla kullanmaları, benimsemeleri ve karşı çıkmamaları için doğru eğitimi ve sürekli desteği sağlamalıdır. Kötü Kullanıcı Deneyimi: Hindistan’da faaliyet gösteren Freshworks adlı bulut tabanlı yazılım şirketi tarafından yapılan araştırma, 10 çalışandan dokuzunun iş yazılımlarından memnun olmadıklarını bildirdi. İşletmelerde kullanılmaya başlanan yeni teknoloji sezgisel veya kullanıcı dostu değilse personellerin yeni teknolojiyi kullanma olasılığı daha düşük olacaktır. Yönetici Desteğinin Olmaması: İşletme personelleri, yöneticilerinden destek ve katılım görmezlerse yeni teknolojiyi benimsemekte tereddüt edebilirler. Yöneticiler yeni teknolojiyi kendileri tanıtmayıp kullanmıyorsa, personeller bu teknolojiyi kullanmanın kendi isteğine bağlı olduğunu veya bu teknolojiyi kullanmanın gereksiz olduğunu düşünebilir. Mevcut Sistemlerle Entegrasyon: Yeni çıkan teknolojilerin işletmenin kullandığı mevcut yazılım veya süreçlerle iyi bir şekilde entegre olmazsa, personeller birden fazla sistem veya geçici çözümler arasında gezinmeye çalışırken verimsizliklere yol açabilir. 5.2 Çalışanları yeni teknoloji konusunda eğitmenin önemi Personellerin bir kısmının bilgi, beceri ve yeterlilik eksikliği olması ve görevi zamanında yerine getirememesi nedeniyle eğitim, işletmelerin gelişimi için bir personel için vazgeçilmez bir unsurdur. Eğitim, bir meslek veya görevin amacına yönelik belirli bilgi ve becerilerin edinilmesine yönelik bir öğrenme faaliyetidir. Eğitimin odak noktası iş veya görevdir; örneğin, belirli makine veya ekipmanın çalışmasında verimlilik ve güvenliğe sahip olma ihtiyacı veya etkili bir satış gücüne duyulan ihtiyaç, bunlardan sadece birkaçıdır (Karim,2019). Personellerin yeni teknoloji konusunda eğitilmesinin önemini destekleyen çeşitli stratejik nedenler aşağıda belirtilmiştir: (E.Flanagan,2023 ve Oluwaseun,2018:185-187). • Müşteri memnuniyetini artırmak • Müşteri memnuniyetini artırmak Yeni teknolojiyle kuruluşlar süreçleri otomatikleştirebilir ve bekleme sürelerini azaltabilir; bu da müşteriler için daha hızlı hizmet ve daha akıcı bir deneyim sağlayabilir. İşyerinde yeni teknolojinin tanıtılması aynı zamanda müşterilere çevrimiçi sohbet robotları, mobil uygulamalar • Artan Verimlilik Personellere zamanında verilen teknolojik eğitim verimliliği artırmaya yardımcı olur. Verilen eğitimler personellerin teknolojik beceri, bilgi ve performans eksikliklerini gidererek ve onların işlerinde hemen uygulamaya koyabilecekleri yeni fikirler, en iyi uygulamalar ve becerilerle donatarak yetkinlik, yetenek ve güven düzeyleri artar (Oluwaseun,2018:185-187). Personellere zamanında verilen teknolojik eğitim verimliliği artırmaya yardımcı olur. Verilen eğitimler personellerin teknolojik beceri, bilgi ve performans eksikliklerini gidererek ve onların işlerinde hemen uygulamaya koyabilecekleri yeni fikirler, en iyi uygulamalar ve becerilerle donatarak yetkinlik, yetenek ve güven düzeyleri artar (Oluwaseun,2018:185-187). • Hızlı bir şekilde ölçeklendirmek • Hızlı bir şekilde ölçeklendirmek İşletmede yeni bir sistem uygulamak, işletmenin hızlı bir şekilde ölçeklenmesine yardımcı olarak operasyonlarını ve gelirlerini hızla genişletmesine imkân tanır. Artan çeviklikle, verimliliği maksimum seviyeye çıkarırken ve genel giderleri minimum seviyeye indirirken müşteri gereksinimlerini daha iyi karşılayabilir, üretkenliği artırarak işletmenin temel yetkinliklerine odaklanabilir ve daha fazla inovasyon sağlayabilir. • Müşteri memnuniyetini artırmak Yeni teknolojiyle kuruluşlar süreçleri otomatikleştirebilir ve bekleme sürelerini azaltabilir; bu da müşteriler için daha hızlı hizmet ve daha akıcı bir deneyim sağlayabilir. İşyerinde yeni teknolojinin tanıtılması aynı zamanda müşterilere çevrimiçi sohbet robotları, mobil uygulamalar 263 Şahin Daştan / Mehmet Göl veya self-servis kiosklar gibi kuruluşla etkileşimde bulunmak için daha fazla seçenek sunarak kolaylık ve erişilebilirliği artırabilir. veya self-servis kiosklar gibi kuruluşla etkileşimde bulunmak için daha fazla seçenek sunarak kolaylık ve erişilebilirliği artırabilir. • Rekabet avantajı kazanmak • Rekabet avantajı kazanmak Teknolojiyi güncel tutmak, kuruluşların rekabette önde gitmesine veya rekabete ayak uydurmasına olanak tanır. Otomasyonun gücünden yararlanarak kuruluşlar daha verimli çalışabilir, bu da maliyetleri azaltabilir ve karlılığı artırabilir. Ek olarak teknoloji, kuruluşların yeni yetenekler sağlayarak yenilik yapmalarını sağlayabilir ve onları rakiplerinden farklılaştıran yeni ürün veya hizmetlere yol açabilir. • Yatırım Getirisi Bir işletme, eğitim ve gelişimi kuruluşun stratejik önceliği haline getirerek ve bunu düşünceli ve bilinçli bir şekilde uygulayarak ölçülebilir yatırım getirisinden yararlanabilir. Etkiyi, niteliksel ve niceliksel yatırım getirisi ölçülebilir. Bu etkiyi ölçmenin çeşitli yolları vardır. Örneğin eğitim öncesi ve sonrası değerlendirmeler yapabilir, gelişimi ölçülebilir. Daha sonra tasarruf edilen zamanın, artan üretkenliğin, artan satışların değeri hesapladığında bunu finansal getiriye dönüştürebilir. İşletmelerin teknolojik eğitim ve gelişimi bir gider değil yatırım olarak görmesi ve iş/kurumsal hedeflere ulaşmak için eğitim ve gelişimden faydalanması önemlidir. Teknolojik yönde personel eğitimi ve gelişimine yapılan yatırım, yatırım getirisi arayacağınız bir sermaye yatırımı olarak görülmelidir. (Oluwaseun,2018:185-187). Bir işletme, eğitim ve gelişimi kuruluşun stratejik önceliği haline getirerek ve bunu düşünceli ve bilinçli bir şekilde uygulayarak ölçülebilir yatırım getirisinden yararlanabilir. Etkiyi, niteliksel ve niceliksel yatırım getirisi ölçülebilir. Bu etkiyi ölçmenin çeşitli yolları vardır. Örneğin eğitim öncesi ve sonrası değerlendirmeler yapabilir, gelişimi ölçülebilir. Daha sonra tasarruf edilen zamanın, artan üretkenliğin, artan satışların değeri hesapladığında bunu finansal getiriye dönüştürebilir. İşletmelerin teknolojik eğitim ve gelişimi bir gider değil yatırım olarak görmesi ve iş/kurumsal hedeflere ulaşmak için eğitim ve gelişimden faydalanması önemlidir. Teknolojik yönde personel eğitimi ve gelişimine yapılan yatırım, yatırım getirisi arayacağınız bir sermaye yatırımı olarak görülmelidir. (Oluwaseun,2018:185-187). 264 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... • Artan Personel Memnuniyeti ve Personeli Elde Tutma Personellerin teknolojik eğitimine ve gelişimine yatırım yapmak, personellerin kendilerini işletmeye daha sadık, değerli, sorumlu, odaklanmış ve ekibin bir parçası hissetmelerini sağlar. Her ne kadar herhangi bir personelin tüm çalışma hayatı boyunca bir işletmede kalmasını beklenmese de bunun personelin sadakat duygusunu artırdığı ve işten ayrılma olasılığını azalttığı kanıtlanmıştır. Birçok insan yapısı gereği yeni şeyler öğrenmek, veri hazinesini ve becerilerini genişletmek ve profesyonel olarak büyümek ister. Çoğu insan çalışma hayatı boyunca ilerlemek ister. İşletme kendi teknolojik gelişimine yatırım yaptığında hem işletme hem de personeller büyük gelişim kazanabilir. Eğitim ve geliştirme sadece bilgi ve beceri boşluklarını doldurmaya yönelik bir araç olmakla kalmaz, aynı zamanda yüksek potansiyele sahip, yüksek başarıya sahip personelleri ödüllendiren bir mekanizmadır. (Oluwaseun,2018:185-187). • İşletmenin Öğrenme Kültürünü Beslemek İşletmenin beklentilerin en başından itibaren açıkça belirlendiği ve hem personel hem de işletme açısından faydaların paylaşıldığı bir öğrenme kültürünü teşvik etmek önemlidir. Organizasyon içindeki başarı hikâyelerine işaret edebilmek ve her zaman işletme hedeflerini gerçekleştirmek önemlidir. İşletme başarılarını garanti altına almak için, bilginin işyerine geri aktarılmasına yönelik bir planın olması önemlidir. Personellerin önemli şeyleri uygulamaya teşvik edilebilir. Personellerin, farklı kuruluşlardaki ve sektörler arası meslektaşlarıyla ağ kurabilecekleri açık kayıt programlarına göndererek, fikirleri güvenli bir ortamda test edebilecekleri ve geri getirebilecekleri fikirleri ve en iyi uygulamaları tartışabilecekleri bir forum sağlanır. Bu yeni fikirleri derhal kendi işletmelerinde uygulamaları ve ayrıca işletmedeki ekipleriyle fikirlerini paylaşmaları teşvik edilmelidir. Bunu yapmanın bir yolu, çalışanı, ekip arkadaşlarına temel çıkarımları sunabilecekleri bir “iş yemeği ve öğrenme” düzenlemeye teşvik etmektir. Bu aynı zamanda bilgi aktarımına da yardımcı olacaktır Çünkü öğrenme kültürünün güçlendirilmesine ve ekip için ortak dil ve çerçevelerin oluşturulmasına yardımcı olacaktır. Günün sonunda en büyük değer personellerdir. İşletmeyi ve işletme teknolojilerini geliştirirken, personellerin bireysel becerilerini yenilememiz, ekiplerimizi geliştirmemiz ve onları rekabetçi kalabilmek ve ellerinden gelenin en iyisini yapabilmek için bilmeleri gereken şeylerle donatmamız gerekiyor. Rekabetin giderek arttığı bu küresel dünyada kazananlar, • İşletmenin Öğrenme Kültürünü Beslemek ş y yg y ş Personellerin, farklı kuruluşlardaki ve sektörler arası meslektaşlarıyla ağ kurabilecekleri açık kayıt programlarına göndererek, fikirleri güvenli bir ortamda test edebilecekleri ve geri getirebilecekleri fikirleri ve en iyi uygulamaları tartışabilecekleri bir forum sağlanır. Bu yeni fikirleri derhal kendi işletmelerinde uygulamaları ve ayrıca işletmedeki ekipleriyle fikirlerini paylaşmaları teşvik edilmelidir. Bunu yapmanın bir yolu, çalışanı, ekip arkadaşlarına temel çıkarımları sunabilecekleri bir “iş yemeği ve öğrenme” düzenlemeye teşvik etmektir. Bu aynı zamanda bilgi aktarımına da yardımcı olacaktır Çünkü öğrenme kültürünün güçlendirilmesine ve ekip için ortak dil ve çerçevelerin oluşturulmasına yardımcı olacaktır. Günün sonunda en büyük değer personellerdir. İşletmeyi ve işletme teknolojilerini geliştirirken, personellerin bireysel becerilerini yenilememiz, ekiplerimizi geliştirmemiz ve onları rekabetçi kalabilmek ve ellerinden gelenin en iyisini yapabilmek için bilmeleri gereken şeylerle donatmamız gerekiyor. Rekabetin giderek arttığı bu küresel dünyada kazananlar, Şahin Daştan / Mehmet Göl \| 265 personellerin eğitimine ve gelişimine yatırım yapan işletmeler olacaktır. (Oluwaseun,2018:185-187). Şahin Daştan / Mehmet Göl 265 personellerin eğitimine ve gelişimine yatırım yapan işletmeler olacaktır. (Oluwaseun,2018:185-187). 5.3 İşletmede Yeni Teknolojilerin Uygulanması İşletmenin herhangi bir yeni teknolojiyi kullanmaya değişim yönetimi stratejiyle uygulamaya başlar. İşletmede yeni teknolojilerin uygulanmaya başlamasında aşağıda belirtilen aşamalar izlenir: E.Flanagan. “How to convince employees to adopt new technology”. Erişim: 01.09.2023. https:// www.unboxedtechnology.com/blog/how-to-implement-new-technology-in- the-workplace/). • Değişim gereksinimlerin belirlenmesi: Değişim yönetiminde ilk aşama değişim gereksinimlerinin belirlenmesidir. Belirlenen değişikliklerin pozitif ve negatif etkilerini değerlendirmektir. • Değişimin planlanması: Değişimi planlarken değişimi hangi sürede gerçekleşeceği ile ilgili zaman çizelgesi, değişimin planlanmasında gerekli kaynakların bir listesi ve net bir iletişim stratejisi eklemelidir. Planda mevcut bulunan engelleri ve bunların nasıl ele alınacağını da tanımlamalıdır. • Personelleri Seçim Süreci Aşamasına Dâhil Etmek: İşletmede hangi teknolojinin kullanılması aşamasında son kararı vermeden önce personellerin seçim sürecine dahil olarak fikirleri alınmalıdır. Personellere karar verme hakkında danışıldığında, personeller kendilerini yeni teknolojiye daha fazla yatırım yapmış hissedeceklerdir. • Personelleri Seçim Süreci Aşamasına Dâhil Etmek: İşletmede hangi teknolojinin kullanılması aşamasında son kararı vermeden önce personellerin seçim sürecine dahil olarak fikirleri alınmalıdır. Personellere karar verme hakkında danışıldığında, personeller kendilerini yeni teknolojiye daha fazla yatırım yapmış hissedeceklerdir. • Yeni Teknolojilerin Faydalarını Vurgulamak: İşletmede yeni teknolojilerin uygulanmasına başladığında yeni teknolojilerin pozitif yönlerini işletme personellerine anlatmak gerekir. Yeni teknolojinin işletmedeki işleri nasıl daha kolay, daha süratli veya daha yararlı hale getireceğini işletmenin personellerine işletme yönetimi tarafından vurgulanmalıdır. • Yeni Teknolojilerin Faydalarını Vurgulamak: İşletmede yeni teknolojilerin uygulanmasına başladığında yeni teknolojilerin pozitif yönlerini işletme personellerine anlatmak gerekir. Yeni teknolojinin işletmedeki işleri nasıl daha kolay, daha süratli veya daha yararlı hale getireceğini işletmenin personellerine işletme yönetimi tarafından vurgulanmalıdır. • Yeni Teknolojinin Kullanması İle İlgili Eğitim Desteği Sağlamak: İşletme personellerine yeni teknolojinin nasıl kullanılacağı konusunda eğitim verilmesi zorunludur. İşletmenin personellerine verilecek bu eğitimler resmi eğitim oturumları, çevrimiçi eğitimler veya bire bir koçluk yoluyla yapılabilir. İşletme personellerinin teknolojiyi verimli bir biçimde kullanmaları için ihtiyaç duydukları yetkinliğe sahip olduklarından emin olunması gereklidir. • Yeni Teknolojinin Kullanması İle İlgili Eğitim Desteği Sağlamak: İşletme personellerine yeni teknolojinin nasıl kullanılacağı konusunda eğitim verilmesi zorunludur. İşletmenin personellerine verilecek bu eğitimler resmi eğitim oturumları, çevrimiçi eğitimler veya bire bir koçluk yoluyla yapılabilir. İşletme personellerinin teknolojiyi verimli bir biçimde kullanmaları için ihtiyaç duydukları yetkinliğe sahip olduklarından emin olunması gereklidir. \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... 266 • Beklentilerin Belirlenmesi: İşletmede Yeni teknolojinin nasıl kullanılacağına ilişkin beklentileri açıkça tanımlanması gereklidir. İşletmede yeni teknolojilerin nasıl kullanılacağı, ne zaman kullanılacağına, nasıl kullanılacağına ve hedeflenen neticelerin ne olduğuna ilişkin yönergeleri içerebilir. Bu yönergeleri personellerin teknolojinin işletmede oynadığı rolü kavramasında yardımcı olacaktır. 5.3 İşletmede Yeni Teknolojilerin Uygulanması • Yeni Teknolojinin Kullanılıp Benimsenmesinde İşletme Yönetiminin Öncü Olması: İşletme yönetimi yeni teknolojiyi kullanmaya ve benimsemeye ilk kendileri başladığında teknolojiye olan sadakatlarını göstererek işletme personellerine nasıl etkili bir şekilde kullanacaklarını göstereceklerdir. 5.3 İşletmede Yeni Teknolojilerin Uygulanması • Beklentilerin Belirlenmesi: İşletmede Yeni teknolojinin nasıl kullanılacağına ilişkin beklentileri açıkça tanımlanması gereklidir. İşletmede yeni teknolojilerin nasıl kullanılacağı, ne zaman kullanılacağına, nasıl kullanılacağına ve hedeflenen neticelerin ne olduğuna ilişkin yönergeleri içerebilir. Bu yönergeleri personellerin teknolojinin işletmede oynadığı rolü kavramasında yardımcı olacaktır. • Yeni Teknolojiyi Kullanıcı Dostu Haline Getirmek: İşletmede uygulanmaya başlayacak teknolojiyi Kullanıcı dostu ve sezgisel teknoloji haline getirilmelidir. Personeller teknolojinin kullanımını kolay buluyorsa, onu benimseme olasılıkları daha yüksektir. • Yeni Teknolojiyi Kullanıcı Dostu Haline Getirmek: İşletmede uygulanmaya başlayacak teknolojiyi Kullanıcı dostu ve sezgisel teknoloji haline getirilmelidir. Personeller teknolojinin kullanımını kolay buluyorsa, onu benimseme olasılıkları daha yüksektir. • Personellere Yeni Teknolojiyi Kullanırken Destek Sağlanması: İş- letme yönetimi tarafından işletmede yeni teknolojiyi kullanırken işlet- me personellerine sürekli destek sağlanması gereklidir. Bu destekler, bir bilişsel teknolojiler yardım masasını, sorun giderme kılavuzlarını veya belirlenmiş bir destek personeline erişimi içerebilir. • Personellere Yeni Teknolojiyi Kullanırken Destek Sağlanması: İş- letme yönetimi tarafından işletmede yeni teknolojiyi kullanırken işlet- me personellerine sürekli destek sağlanması gereklidir. Bu destekler, bir bilişsel teknolojiler yardım masasını, sorun giderme kılavuzlarını veya belirlenmiş bir destek personeline erişimi içerebilir. • Personelleri Yeni Teknolojileri Denemelerine Teşvik Etmek: İşletme personellerine yeni teknolojilere denemelerine teşvik edilmesi, yeni teknolojilerin kullanmanın yeni yollarını keşfetmelerine ve rahat olmalarına yardımcı olacaktır. • Personelleri Yeni Teknolojileri Denemelerine Teşvik Etmek: İşletme personellerine yeni teknolojilere denemelerine teşvik edilmesi, yeni teknolojilerin kullanmanın yeni yollarını keşfetmelerine ve rahat olmalarına yardımcı olacaktır. • İşletme Yönetimi Tarafından Başarının Kutlanması: İşletme yönetimi tarafından işletme personelleri tarafından yeni teknolojinin kabul edilmesi halinde başarıların kutlanması, teknolojiyi benimseyen kabul eden personellerin takdir edilmesini ve başarı hikâyelerini ekibin diğer üyeleriyle paylaşılmasını içerebilir. • İşletme Yönetimi Tarafından Başarının Kutlanması: İşletme yönetimi tarafından işletme personelleri tarafından yeni teknolojinin kabul edilmesi halinde başarıların kutlanması, teknolojiyi benimseyen kabul eden personellerin takdir edilmesini ve başarı hikâyelerini ekibin diğer üyeleriyle paylaşılmasını içerebilir. • Yeni Teknolojiyi Kabul Etmeyip Direnenlere Karşı Mücadele Etmek: İşletme yönetimi yeni teknolojiye karşı olan her türlü direnişi ele alıp, personellerin endişelerini gidererek, onlara ek eğitim sağlayarak veya teknik sorunları ele alarak yardımcı olunur. • Yeni Teknolojiyi Kabul Etmeyip Direnenlere Karşı Mücadele Etmek: İşletme yönetimi yeni teknolojiye karşı olan her türlü direnişi ele alıp, personellerin endişelerini gidererek, onlara ek eğitim sağlayarak veya teknik sorunları ele alarak yardımcı olunur. • Yeni Teknolojinin Kullanılıp Benimsenmesinde İşletme Yönetiminin Öncü Olması: İşletme yönetimi yeni teknolojiyi kullanmaya ve benimsemeye ilk kendileri başladığında teknolojiye olan sadakatlarını göstererek işletme personellerine nasıl etkili bir şekilde kullanacaklarını göstereceklerdir. 6. Sonuç Başarılı bir muhasebe meslek mensubu olmak için gerekli teknoloji yetkinlikleri sürekli geliştirmek gerekir. Zaman ilerledikçe değişen sektör şartlarıyla birlikte gelişim gösterebilmek için teknolojik açıdan yeterli olmak 267 Şahin Daştan / Mehmet Göl \| gerekir. Muhasebe meslek mensuplarının ofislerinde giderek daha fazla teknoloji kullanımına öncelik verdikçe, verilerin bulut tabanlı sistemlere aktardıkça ve dijital süreci iş akışlarına dâhil ettikçe, muhasebe mesleğinin gerektirdiği geleneksel bilgi ve becerileri korurken bir yandan da çeşitli teknolojik alanlarda yetkin hale gelmeleri gerekmektedir. Muhasebe meslek mensuplarına, stratejik düşünme, sürdürülebilirliği ele alma, yenilikçiliği destekleme ve dijital gündemi yönlendirme yetkinleri ile birlikte ilerleyen zamanda kilit roller düşecektir. Muhasebe meslek mensuplarına yakıştırılan beş geniş rol alanı aşağıda belirtilmiştir: ACCA Global.“Future accountants: a bright future awaits”.Erişim: 05.09.2023. https://yourfuture.accaglobal. com/global/en/blog/Future-accountants-a-bright-future-awaits.html Güvence savunucusu: Dijital olarak kesintiye uğrayan ve hızla değişen sektörlerde, kuruluşların riski uygun ve etik bir şekilde yönetmeleri gerekmektedir. Profesyonel muhasebe meslek mensupları, değer yaratmak ve korumak amacıyla risk, raporlama ve iç kontrol ortamını dönüştürmek için dijital araçları ve teknolojileri kullanabilir. Ayrıca sorumlu iş uygulamalarını destekleyebilir ve kuruluşta şeffaflığı ve güveni artırabilirler. İş transformatörü: Günümüzde iş modelleri, yeni teknolojilere ve pazarlara yanıt olarak gelişmeye devam edecektir. Muhasebeciler, finans operasyonel değişim programlarına veya daha geniş iş dönüşümü girişimlerine liderlik ederek bu çabalarda her büyüklükteki işletmeyi destekleyebilir. Veri gezgini. Günümüzde veri, yeni yatırımlar için iş gerekçesi oluşturmaktan rekabetçi tehditlerin profilini çıkarmaya kadar her alanda kullanılabilen kurumsal bir varlıktır. Muhasebeciler, uzun vadeli değerin nasıl yaratılacağı ve sürdürüleceği konusunda gerçek zamanlı analizler sağlamak için zengin veri setlerinin ve analitik araçların geliştirilmesine ve uygulanmasına yardımcı olarak stratejik danışmanlar olarak hareket edebilirler. Dijital oyun kurucu. Dijital uyum, inovasyon yoluyla rekabet avantajı yaratmanın anahtarıdır. Muhasebe meslek mensuplarının, finans ve iş operasyonlarını dönüştürmek için robotik ve makine öğreniminin potansiyelini belirleyerek ve üretkenliği ve daha iyi karar desteğini artırmak için teknoloji ekipleriyle birlikte çalışarak teknoloji savunucuları olarak hareket edebilirler. Sürdürülebilirliğin öncüsü. Günümüzde kaynakların giderek kısıtlandığı ve iş modellerinin bozulduğu bir dünyada, sadece finansal sermayeden daha fazlası değer yaratmakta ve maddi olmayan varlıklar kurumsal değerin giderek artan bir kısmını oluşturmaktadır. Muhasebe meslek mensupları, gerçekten değer yaratan faaliyetleri yakalayan, ölçen ve raporlayan çerçeveler belirleyerek kuruluşun performansı hakkında daha anlamlı ve şeffaf bilgiler sağlayabilir. Dijital Muhasebe – Modern Eğitim – Yetkin Meslek Mensubu 268 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... Kaynakça: ACCA Global.“Future accountants: a bright future awaits”.Erişim: 05.09.2023. https://yourfuture.accaglobal.com/global/en/blog/Futu- re-accountants-a-bright-future-awaits.html Aksu, G.S. ve Dalkılıç-Sürgevil, O. 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Özet Küçük ve Orta Ölçekli İşletmeler (KOBİ’ler) ekonomik büyümenin sağlanmasında ve yenilikçiliğin teşvik edilmesinde önemli bir rol oynasa da genellikle pazarlama ve finans alanlarında sürdürülebilir kalkınmalarını engelleyen ciddi zorluklarla karşılaşmaktadır. Pazarlama alanında, KOBİ’ler sınırlı kaynaklar ve ekonomik değişimlerle birlikte niteliğe dayalı pazarlama eksikliği ve dijital pazarlamanın gelişen manzarası gibi birtakım engellerle karşılaşmaktadır. Finans alanında ise KOBİ’ler yetersiz sermaye, finansal teknik bilgi ve okuryazarlık seviyesi düşüklüğü, ekonomik istikrarsızlık ve teşvik kaynaklı sorunlar gibi sorunlar ile yüzleşebilmektedir. Bu çalışma ile Düzce ilinde faaliyet gösteren KOBİ’lerin pazarlama ve finans sorunları açısından incelenerek genel bir tablonun ortaya çıkarılması ve söz konusu faaliyetlerine dayalı sorunlarına çözüm önerileri sunarak hem literatüre hem de KOBİ’lere bir çıktı sunmak ve paylaşmak amaçlanmıştır. Pazarlama ve finans arasındaki etkileşim KOBİ’lerin hayatta kalabilmesi için büyük önem arz etmektedir ve bu çalışma ile, birbirine entegre bir yaklaşıma duyulan ihtiyaç vurgulanmıştır. Ayrıca bu çalışma KOBİ’lerin karşılaştığı pazarlama ve finansal zorluklar arasındaki karmaşık ilişkiyi vurgulayarak söz konusu sorunları ele almada bütüncül bir yaklaşım önermektedir. Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme Serhat Ata1 Ali Özer2 Nevin Özer3 Serhat Ata1 Ali Özer2 Nevin Özer3 Kaynakça: sciencedirect.com/journal/computers-and-education/vol/68/suppl/C”- Volume 68, October 2013, Pages 473-481 https://doi.org/10.1016/j. compedu.2013.06.008 Gulin, D., Hladika, M. and Valenta, I. (2019).“ “Digitalization and the Chal- lenges for the Accounting Profession””. in Proceedings of the ENTRE- NOVA - ENTerprise REsearch InNOVAtion Conference, Rovinj, Croa- tia, 12-14 September 2019, IRENE - Society for Advancing Innovation and Research in Economy, Zagreb, pp. 502-511. https://www.econstor. eu/bitstream/10419/207712/1/56-ENT-2019-Gulin-et-al-502-511.pdf London School of Business & Finance. “How Will Technology Change Ac- counting in the Future?” Erişim: 28.08.2023. https://www.lsbf.org.uk/ blog/online-learning/how-will-technology-change-accounting-in-the-fu- ture ): Karim, M., Latif,B,W., Choudhury,M,M. (2019) . “The impact of training and development on employees performance: An analysis of quanti- tative data”. Noble International Journal of Business and Management Research ISSN(e): 2520-4521 ISSN(p): 2522-6606 Vol. 03, No. 02, pp: 25-33, 2019. https://www.researchgate.net/publication/331147751 Makaruk, P.,(2021) . “New skills of the accountant profession in the di- gital economy”. pp. 131-133. http://edoc.bseu.by:8080/bitstream/ edoc/90845/1/Makaruk%20131-133.pdf 270 \| Dijitalleşen Muhasebe Dünyasına Meslek Mensuplarının Uyumu Açısından Mesleki Yetkinlik... Mancini, D, Lamboglia,R., Gastellona,N.G., And Corsi.K.(2017). “Trends of Digital Innovation Applied to Accounting Information and Mana- gement Control Systems”. Reshaping Accounting and Management Control Systems, Part of the Lecture Notes in Information Systems and Organisation book series (LNISO,volume 20).Editors Corsi,K., Gastellona, N.G, Lamboglia,R., Mancini,D. pp. 1-19. https://link. springer.com/chapter/10.1007/978-3-319-49538-5_1 Oluwaseun,O,O.(2018) . “ Employee training and development as a mo- del for organizational success”. International Journal of Engineering Technologies and Management Research, 5(3), 181–189. https://doi. org/10.29121/ijetmr.v5.i3.2018.190 Protech Service Group. “5 Benefits of Technology Training”. Erişim: 28.08.2023. https://www.psgi.net/5-benefits-of-technology-training/ PwC (2017). Global Digital IQ Survey 2017. https://www.pwc.com/sk/en/ publikacie/assets/2017/pwc-digital-iq-report.pdf Tekbaş. İ.“Gelecekte bir muhasebecinin ihtiyaç duyacağı 6 be- ceri”. Erişim: 28.08.2023 https://www.parasut.com/blog/ bir-muhasebecinin-bilmesi-gerekenler) Bölüm 15 1 Araştırma Görevlisi, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-5423-5118 2 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0003-4736-3418 3 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-1736-4199 2 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0003-4736-3418 3 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-1736-4199 1 Araştırma Görevlisi, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-5423-5118 2 Doçent Doktor Düzce Üniversitesi İşletme Fakültesi ORCID: 0000-0003-4736-3418 1 Araştırma Görevlisi, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-5423-5118 3 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0002-1736-4199 2 Doçent Doktor, Düzce Üniversitesi İşletme Fakültesi, ORCID: 0000-0003-4736-3418 Giriş Küçük ve Orta Ölçekli İşletmeler (KOBİ’ler), çoğu ekonomide, özellikle de gelişmekte olan ülkelerde önemli bir rol oynamaktadır. KOBİ’ler dünya https://doi.org/10.58830/ozgur.pub401.c1763 https://doi.org/10.58830/ozgur.pub401.c1763 271 272 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme çapındaki işletmelerin çoğunluğunu oluşturmaktadır ve iş yaratma ve küresel ekonomik kalkınmaya önemli katkılarda bulunmaktadır. Birçok ülke, çeşitli program ve politikalarda KOBİ’lere yönelik destek uygulamaktadır. Çünkü KOBİ’ler daha yüksek istihdam artışı, çıktı, ihracatın teşviki ve girişimciliği teşvik etme gibi çeşitli sosyo-ekonomik hedeflerin tatmin edilmesine önemli katkısı nedeniyle dünya çapında ülkelerce iyi bilinmektedir (Keskin vd., 2010). KOBİ’ler geliştirilmesi, merkezi bir tema olarak sürdürülebilir büyümenin sağlanmasına yardımcı olmaktadır. KOBİ’ler ülkenin genel üretim ağlarında hayati bir rol oynamaktadır ve gelişmekte olan ülkelerin ekonomik büyümesinin merkezinde yer almaktadır. Kayıtlı KOBİ’lerin katkıları, toplam istihdamın %50’si ve gelişmekte olan ekonomilerin milli gelirinin %33’üdür. Kayıt dışı KOBİ’ler dâhil edilince söz konusu yüzdeler çok daha fazla olacaktır. Finansman sorunu KOBİ’lerin büyümesinin önündeki en büyük engeldir (Maarg, 2016). Finansman ile doğası gereği ilişkili olarak pazarlama sorunları da KOBİ’lerin önünde en önemli konular arasında yer almaktadır ve çünkü bir KOBİ’nin başarılı olup olmayacağı uzun vadede ürün veya hizmetlerini ne kadar pazarlayabildiğine bağlıdır (Scheers, 2011). Söz konusu sorunlar derinlemesine irdelendiğinde, KOBİ’lerin kendi denetimleri haricinde meydana gelen çıktılardan oldukça etkilendikleri görülmektedir. Her ne kadar KOBİ’ler birtakım zorluklarla karşı karşıya gelse de, birçok ekonomi, özellikle istihdam yaratmak için KOBİ’lere büyük ölçüde bağımlıdır. Zayıflıklarına rağmen, KOBİ’ler küreselleşme ve bölgesel bütünleşme süreci ile yok olma eğilimi göstermemiştir, daha ziyade rolleri ve katkıları değişmiş ve gelişmiştir (Harvie ve Charoenrat 2015). 1. Küçük Ve Orta Büyüklükteki İşletmeler (Kobi) Tanımı KOBİ’ler, iki yüz elli kişiden az çalışan istihdam eden ve yıllık net satış hasılatı veya mali bilançosundan herhangi biri beş yüz milyon Türk Lirasını aşmayan ekonomik birimler veya girişimler olarak tanımlanmaktadır (Küçük ve Orta Büyüklükteki İşletmeler Yönetmeliği 2023). Avrupa Birliği (2021), küçük işletmeyi 50’den az çalışanı olan ve orta ölçekli işletmeyi 250’den az çalışanı olan bir işletme olarak tanımlamaktadır. Küçük işletmeler ABD’de, genellikle 100’den az çalışanı olan bir işletme olarak tanımlanmaktadır. 10’dan az çalışanı olan bir kuruluş, genellikle küçük bir ofis/ev ofis (SOHO) veya bir mikro işletme olarak sınıflandırılmaktadır (ABD Küçük İşletmeler İdaresi, 2022). Ülkeler, KOBİ’lerin ekonomiye olan faydalarını geliştirmek adına birtakım politika ve programları yürütmeye çalışmakta ve politika ve programların 273 Serhat Ata / Ali Özer / Nevin Özer \| hedef kitlesini saptayabilmek maksadıyla da kendi KOBİ tanımlarını düzenlemektedir (Cansız, 2008). Çeşitli tanımlamalardan anlaşılacağı üzere Türkiye de dâhil, KOBİ’nin evrensel olarak kabul görmüş bir tanımı yoktur. Her ülkenin küçük ve orta ölçekli bir işletmeyi neyin oluşturduğuna dair kendi tanımı vardır. Belirli büyüklük şartlarının karşılanması gerekir ve bazen firmaların faaliyet gösterdiği sektör de dikkate alınmaktadır. Scheers’ e (2011) göre, küçük bir işletmenin evrensel bir tanımını formüle etmek zordur çünkü ülkelerin ekonomileri farklıdır ve insanlar belirli amaçlar için belirli standartları benimsemektedir. Fatai’ ye (2011) göre, KOBİ’leri tanımlamak için genel kullanılan kriterler “çalışan sayısı, yıllık ciro, yerel operasyonlar, satış hacimleri, finansal güç, yöneticiler ve özerklik seviyesi, nispeten küçük pazarlar ve genellikle bireyler tarafından sağlanan sermaye olarak sıralanabilir. Tablo 1 Dünya’da ve Türkiye’de çeşitli kriterler temel alınarak ele alınan KOBİ tanımlarını göstermektedir. Tablo 1. 1. Küçük Ve Orta Büyüklükteki İşletmeler (Kobi) Tanımı Farklı Kurum ve Kuruluşlar açısından KOBİ tanımları Kurum ve Kuruluşlar Kategori İşçi Sayısı Net Satış Hasılatı(yıllık) Sektör Avrupa Komisyonu (EC, 2003) Mikro <10 2 milyon € Sektör Ayrımı Yok Küçük <50 10 milyon € Orta <250 50 milyon € Ekonomik Kalkınma ve İşbirliği Örgütü (OECD, 2005) Küçük 10-49 10 milyon € Sektör Ayrımı Yok Orta 50-249 50 milyon € Birleşmiş Milletler Avrupa Ekonomik Komisyonu (UNECE,2020) Mikro 1-9 1-2 milyon € Sektör Ayrımı Yok Küçük 10-49 5-10 milyon € Orta 50-249 50 -70 milyon € Amerika Birleşik Devletleri Uluslararası Ticaret Komisyonu (USTIC,2010) Küçük <500 ≤7 milyon $ İmalat Orta <500 ≤25 milyon $ Hizmet Küçük ve Orta Ölçekli İşletmeleri Geliştirme ve Destekleme İdare Başkanlığı (KOSGEB, 2023) Mikro <10 10 milyon ₺ Sektör Ayrımı Yok Küçük <50 100 milyon ₺ Orta <250 500 milyon ₺ Türkiye İstatistik Kurumu (TUİK,2020) Mikro <10 5 milyon ₺ 50 milyon ₺ Sektör Ayrımı Yok Küçük <50 Orta <250 250 milyon ₺ Kaynak: Yazarlar tarafından derlenmiştir. Tablo 1. Farklı Kurum ve Kuruluşlar açısından KOBİ tanımları Kaynak: Yazarlar tarafından derlenmiştir. İşletmelerin uluslararası pazarlarda rekabet edebilme yeteneği, iç piyasada varlığını sürdürebilmesi adına sahip olması gereken minimum kriterler ve ülkelerin KOBİ’lere sağlayacağı imkân ve kaynakların destek programları 274 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme içindeki yeri tanımlamalardaki belirleyici temel niteliklerdir (Cansız, 2008). Tablo 1’ deki farklı tanımlamalar KOBİ’lerin birçok farklı şekil ve boyutta ortaya çıktığı; ancak günümüzün karmaşık iş ortamında diğer kuruluşlarla yakın finansal, operasyonel veya yönetimsel ilişkilere sahip olabileceğini ve bu ilişkilerin genellikle bir KOBİ ile daha büyük bir işletme arasındaki çizgiyi kesin olarak çizmeyi zorlaştırdığını göstermektedir (Avrupa Birliği Komisyonu, 2020). Türkiye’de KOBİ ile ilgili resmi olarak en son tanımlama 30.04.2018 tarih ve 11828 karar sayılı Resmi Gazete’de yayımlanarak yürürlüğe giren, Nitelikleri ve Sınıflandırılması Hakkında Yönetmelik ile KOBİ tanımı ve sınıflandırmasıdır. Bu tanıma göre ‘‘Küçük ve Orta Büyüklükte İşletme (KOBİ), ikiyüzelli kişiden az yıllık çalışan istihdam eden ve yıllık net satış hasılatı veya mali bilançosundan herhangi biri yüz yirmi beş milyon Türk Lirasını aşmayan ve bu yönetmelikte mikro işletme, küçük işletme ve orta büyüklükteki işletme olarak sınıflandırılan ekonomik birimleri veya girişimleridir” Son olarak 2022 yılı içerisinde 18.03.2022 tarihli 5315 karar sayılı ‘’Küçük ve Orta Büyüklükteki İşletmelerin Tanımı, Nitelikleri ve Sınıflandırılması Hakkında Yönetmelikte Değişiklik Yapılmasına Dair Yönetmelik’’ yürürlüğe girerek KOBİ tanımında kullanılan kıstaslar güncellenmiştir. 1. Küçük Ve Orta Büyüklükteki İşletmeler (Kobi) Tanımı Buna göre; 250 kişiden az çalışan istihdam eden, yıllık net satış hasılatı veya mali bilançosundan herhangi biri 250 milyon lirayı aşmayan işletmeler KOBİ olarak tanımlanmıştır. İlaveten, 10 kişiden az çalışanı olan mikro işletmelerin yıllık net satış hasılatı veya mali bilançosu 3 milyon liradan 5 milyon liraya çıkartılmış, 50 kişiden az personeli olan küçük işletmeler için limit, 25 milyon liradan 50 milyon liraya yükseltilmiştir. Düzenleme ile 250’den az çalışanı olan orta büyüklükteki işletmeler için üst limit 125 milyon lirayken 2 katına çıkartılarak 250 milyon lira olarak güncellenmiştir (KOSGEB, 2022). 24.05.2023 tarihli 32201 sayılı resmi gazete de çıkan “Küçük ve Orta Ölçekli İşletmeler Yönetmeliği” ile 2022 yılında çıkan yönetmelik kaldırılmış ve KOBİ’lerin tanımında yer alan limitler 2 katına çıkarılmıştır. Kanuna göre; KOBİ’lerin tanımında yer alan mali bilanço verisi beş yüz milyon Türk Lirasını aşmayan işletmeler olarak güncellenirken, mikro işletmeler için bu rakam on milyon Türk Lirasına, küçük işletmeler için yüz milyon Türk Lirasını, orta büyüklükteki işletmeler için ise beş yüz milyon Türk Lirasını aşmayan işletmeler olarak güncellenmiştir. Serhat Ata / Ali Özer / Nevin Özer \| 275 Tablo 2. KOBİ Tanımı Yönetmeliğine Göre İşletme Sınıflandırılması Ölçek Çalışan Sayısı Net Satış Hasılatı veya Mali Bilanço (TL) Mikro 0-9 10.000.000 Küçük 10-49 100.000.000 Orta 50-249 500.000.000 Büyük 250+ 500.000.000 TL ve fazlası Kaynak: 25 Mayıs 2023 tarihli 32201 sayılı Resmi Gazete Tablo 2. KOBİ Tanımı Yönetmeliğine Göre İşletme Sınıflandırılması 1.1. Kobilerin Dünya ve Türkiye Ekonomisindeki Yeri Küçük ve orta boy işletmelerin (KOBİ) ülke ekonomileri dâhilindeki ehemmiyeti gün geçtikçe artmakta ve bu işletmelerin istihdam, üretim, ihracat, katma değer ve yatırım içindeki payları, önemli büyüklüklere erişmektedir. Kobi’ler olmadan dünyadaki hiçbir ekonominin gelişimi sağlanamaz. Çünkü sayıları büyük firmalardan önemli ölçüde fazladır, çok sayıda insanı istihdam eder, genellikle doğaları gereği girişimcidir ve yeniliğin şekillenmesine yardımcı olurlar (Muritala ve Awolaja ve Bako, 2012). 2020’de dünya çapında yaklaşık 212,98 milyon Kobi olduğu tahmin edilmiştir. Gelişmekte olan ekonomilerdeki birçok kişi KOBİ’ler iş bulmaktadır. Ekonomik İşbirliği ve Kalkınma Örgütü’ne (OCED) göre, KOBİ’ler gelişmekte olan ülkelerde toplam istihdamın yaklaşık %50’sine ve GSYİH’nın %40’ına katkıda bulunmaktadır (World Bank, 2022). Son yirmi yılda da, küçük ve orta ölçekli işletmelerin (KOBİ’ler) iş yapabilme potansiyelleri keşfedildiğinden uluslararasılaşmasına olan ilgi hızla artmıştır (Ribau, Moreira & Raposo, 2018). Ekonomik Kalkınma ve İşbirliği Örgütü (2017) KOBİ’lerin, dijitalleşen küresel ekonomide katkısının artırılmasına yönelik raporunda önemini şu şekilde sıralamıştır; • KOBİ’ler daha kapsayıcı küreselleşme ve büyüme sağlamak için gereklidir. KOBİ’ler ekonomide ve firmaların daha geniş eko- sistemindeki kilit oyunculardır. KOBİ’ ler, uyum sağlamak, daha açık bir ortamda gelişmek, dijital dönüşüme daha aktif katılmak ekonomik büyümeyi artırmak, daha kapsayıcı bir küreselleşme ortamı için gereklidir. • KOBİ’ler, ekonomik ve sosyal refaha daha da geliştirilebilecek çeşitli katkılarda bulunurlar. KOBİ’ler, istihdam ve katma değer yaratarak ve inovasyona katkıda bulunarak dünya çapında ulusal ekonomilerde kilit bir rol oynamaktadır. 276 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme • KOBİ’ler, Dünya genelinde çevresel sürdürülebilirliği ve daha kapsayıcı büyümeyi sağlama çabalarının merkezinde yer almaktadır. Ancak, bu katkılar firmalar, ülkeler ve sektörler arasında büyük farklılıklar göstermektedir. Küresel pazarlara ve bilgi ağlarına daha iyi erişim, KOBİ’lerin katkılarını güçlendirebilir, ancak ticaret ve yatırım engelleri KOBİ’lerin katılımını azaltmakta ve uluslararası pazarlarda rekabetçi maliyetlerle birlikte zayıf fiziksel yapı ve BİT altyapısı, KOBİ’lerin verimli bir şekilde çalışmasını ve erişimini engellemektedir. • İş ortamı, KOBİ’lerin açık ve entegre bir ekonomiye katılımını artırmak ve bundan faydalanmak için kritik öneme sahiptir. Kurumsal ve düzenleyici çerçevenin belirli özellikleri, KOBİ’ler üzerinde orantısız yüklere yol açmaktadır. Verimsiz borçlanma, iş dinamizmini, sürdürülebilir firmaların yeniden yapılandırılmasını ve KOBİ’lerin dış finansmana erişimini sınırlamaktadır. Vergi uyumunun yüksek maliyetleri, küçük ve genç firmalara orantısız bir şekilde düşmektedir. Kamu sektörü bütünlüğü ve şeffaflığı, kamu yönetiminin verimliliği ve kamu kalitesi hizmetler, eşit bir rekabet için gereklidir. • Stratejik kaynaklara erişim, KOBİ’lerin rekabet edebilirliği için kritik öneme sahiptir. Birçok yeni işletme ve KOBİ için, uygun biçimlerde finansmana erişim, bir dizi talep ve arz yönlü sınırlamalarla engellenmektedir. Tablo 3. Büyüklük işletmelere göre KOBİ’lerin temel kriterlerinin oransal dağılımı (%), 2020 1.1. Kobilerin Dünya ve Türkiye Ekonomisindeki Yeri 2020 yılında yaklaşık 3,2 milyon olan KOBİ sayısı, 2021 yılında 3,4 milyon olarak kayıtlara geçmiştir. Tablo 5. Türkiye’deki KOBİ’lerin Çalışan Başına Katma Değer (2009-2020) Kaynak: TUİK KOBİ Girişim İstatistikleri (2021) Tablo 5. Türkiye’deki KOBİ’lerin Çalışan Başına Katma Değer (2009-2020) Kaynak: TUİK KOBİ Girişim İstatistikleri (2021) Tablo 5’te çalışan başı katma değer istatistiklerine bakıldığında, bütün büyüklükteki işletmelerin çalışanlarından elde ettiği katma değerde ciddi bir artış olduğu görülmektedir. ancak işletmelerin büyüklüğü arttıkça katma değer veriminin daha yüksek olduğu görülmektedir. Orta boy işletmeler çalışan başına 173 Bin TL katma değer elde ederken, mikro işletmelerde kişi başı 28 Bin TL katma değer elde edilebilmiştir. 1.1. Kobilerin Dünya ve Türkiye Ekonomisindeki Yeri Beceri eksiklikleri, kötü yönetim uygulamaları ve işgücü eğitimi, KOBİ’lerin üretkenliğini ve yenilikçiliğini sınırlamaktadır. • KOBİ’ler konusunda ortak bir bakış açısına ihtiyaç vardır. KOBİ’ler tipik olarak iş ekosistemlerine büyük şirketlerden daha bağımlıdır ve iç kısıtlamaları nedeniyle piyasa başarısızlıklarına, politika verimsizliklerine ve tutarsızlıklara karşı daha savunmasızdır. Yapısal reformların KOBİ iş ortamı üzerindeki birleşik etkilerinin yanı sıra, inovasyonu teşvik etmek, iş dinamizmini sürdürmek, becerilerin gelişimini desteklemek, KOBİ’lere erişimi kolaylaştırmak da dahil olmak üzere KOBİ’leri hedefleyen politikaların rolü ve etkisi üzerinde daha iyi anlamak için büyük bir çaba gerekmektedir (OECD, 2017). TÜİK (2021) verilerine göre, sanayi ve hizmet sektörlerinde faaliyet gösteren 3 milyon 295 bin girişim KOBİ olarak ifade edilmiştir. Türkiye’de KOBİ’ler 2020 yılında toplam girişim sayısının %99,8’ini oluşturmuştur. Buna karşılık; istihdamın %72’sini, personel maliyetinin %49,2’sini, cironun %49,4’ünü, üretim değerinin %42,7’sini ve faktör maliyetiyle katma değerin %41,3’ünü meydana getirmiştir (TUİK,2021). KOBİ’lerde girişimler sektörlere göre incelendiğinde, aktif olan girişimlerin en çok “toptan ve TÜİK (2021) verilerine göre, sanayi ve hizmet sektörlerinde faaliyet gösteren 3 milyon 295 bin girişim KOBİ olarak ifade edilmiştir. Türkiye’de KOBİ’ler 2020 yılında toplam girişim sayısının %99,8’ini oluşturmuştur. Buna karşılık; istihdamın %72’sini, personel maliyetinin %49,2’sini, cironun %49,4’ünü, üretim değerinin %42,7’sini ve faktör maliyetiyle katma değerin %41,3’ünü meydana getirmiştir (TUİK,2021). KOBİ’lerde girişimler sektörlere göre incelendiğinde, aktif olan girişimlerin en çok “toptan ve 277 Serhat Ata / Ali Özer / Nevin Özer \| 277 perakende ticaret; motorlu kara taşıtlarının ve motosikletlerin onarımı” sektöründe yer aldığı görülmektedir. KOBİ’lerde üretim değeri en fazla olan sektörün 993 milyar TL üretim değeriyle “imalat”sektörüdür. Toplam ihracatın ise yüzde 56,3’ünü KOBİ’ler gerçekleştirmiştir. İhracatta en büyük pay ticaret sektörünündür (TOBB, 2020). Tablo 3. Büyüklük işletmelere göre KOBİ’lerin temel kriterlerinin oransal dağılımı (%), 2020 Tablo 3. Büyüklük işletmelere göre KOBİ’lerin temel kriterlerinin oransal dağılımı (%), 2020 Kaynak: TUİK, 2020: Küçük ve Orta Büyüklükteki Girişim İstatistikleri y ş g ğ (%), 2020 Kaynak: TUİK, 2020: Küçük ve Orta Büyüklükteki Girişim İstatistikleri Kaynak: TUİK, 2020: Küçük ve Orta Büyüklükteki Girişim İstatistikleri Kaynak: TUİK, 2020: Küçük ve Orta Büyüklükteki Girişim İstatistikleri Tablo 4. Türkiye’deki İşletmelerin Oransal Gösterimi (%), 2021 Tablo 4. Türkiye’deki İşletmelerin Oransal Gösterimi (%), 2021 Kaynak: TUİK KOBİ Girişim İstatistikleri (2021) Kaynak: TUİK KOBİ Girişim İstatistikleri (2021) 278 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme Türkiye’deki işletmeler incelendiğinde, toplam işletmelerin %99,7’si mikro, küçük ve orta olarak sınıflandırılan KOBİ’lerden oluştuğu görülmektedir. Büyük işletmeler ise toplam işletmelerin %0,3’ünü oluşturmaktadır. 2. Kobiler Açısından Pazarlama ve Sorunları Ekonomide ve sanayileşmede katma değer oluşturan KOBİ’lerin, çare bulunması hususi olan pek çok sorunu bulunmaktadır. KOBİ’ler artan rekabet, hızla değişen pazar talebine uyum sağlama yeteneği, teknolojik değişim ve bilgi, yenilik ve yaratıcılıkla ilgili kapasite kısıtlamalarından kaynaklanan zorluklarla karşı karşıyadır (ADBInstitue, 2016). Bu problemlerin bir bölümü de işletmelerin kendi niteliklerinden kaynaklanmaktadır (Müftüoğlu, 1991). KOBİ’ler; finansman, üretim, planlama, pazarlama, yönetim, denetim ve dış ticaret gibi alanlarda çok yönlü sorunlarla karşı karşıya gelmektedirler. Pazarlama sorunları KOBİ’lerin başarısını engelleyen en önemli nedenler arasında yer almaktadır ve KOBİ’ler için ortaya çıkan önemli bir sorundur çünkü 279 Serhat Ata / Ali Özer / Nevin Özer \| pazarlama bir KOBİ’nin başarılı olup olmayacağını uzun vadede belirlemektedir (Scheers, 2011). Sorunun ana hatları çoğunlukla KOBİ sahiplerinin ürünlerini genişletemedikleri ve ürün/hizmet kalitesini koruyamadıkları için KOBİ’ler arasındaki yüksek başarısızlık oranları ile açıklanmaktadır (Arasti, Zandi ve Bahmani, 2014). KOBİ’lerin karşılaştıkları pazarlama sorunları genel hatlarıyla; ekonomik koşullardaki değişimlere kısa sürede uyum gösterilememesi, çevre ve pazar analizi eksikliği, yeni pazarlama kanallarının oluşturulamaması, dağıtım kanalları sorunları, pazarlama bilgisi eksikliği, yoğun rekabet ve rekabet edilebilir fiyatlandırma konusundaki sorunlar ve yeni teknolojiye adapte olamama olarak sıralanabilir (Korkmaz, 2003). Cant (2012) KOBİ’lerin karşı karşıya olduğu pazarlama sorunlarını pazar ve büyüme potansiyeli dayalı anlayış eksikliği, pazar bölümlendirme eksikliği, pazar ihtiyaç analizi eksikliği, pazarlama eylemlerini finanse etmek için finansmana erişim eksikliği, eğitimli personel eksikliği, rekabetçiliğe dayalı eksiklikler ve ürünlerin pazarlanması ve sonrası hizmetlere dayalı eksiklikler olarak sıralamıştır. Erdem (2019) ise KOBİ’lerin pazarlama hususundaki zayıf yönlerini; • KOBİ’lerin üretim fonksiyona dayalı bütçesinden kaynaklanan yetersiz pazarlama bütçesi • Hedef pazar araştırması, bulma ve değerlendirmeyle ilişkili tahminlere dayanmayan, sezgiye ve tecrübeye dayalı pazar bilgisi eksikliği • Değişen şartlara ve oluşan fırsatlara cevap verebilecek pazarlama planlaması eksikliği • KOBİ’lerin bürokratik ve ekonomik nedenlerle birlikte rekabet düzeyinin yetersiz kalması ile yerel ve bölgesel pazarlarda pazarlama politikası benimsemesi • KOBİ’lerin hem bilgi hem de alt yapı eksikliğinden dolayı e-ticaretten yeterli düzeyde faydalanamaması • Teknolojik değişim ve gelişmeleri hem de müşteri talepleri göz önünde bulundurmamaktan kaynaklı etkili bir hizmet ve satış sonrası hizmetlerini yeterli düzeyde sunamama ve müşteri memnuniyeti sağlayamama • KOBİ’nin pazarlama karması elemanlarından kaynaklanan sorunları; ürün, fiyat, dağıtım ve tutundurma gibi problemler olarak sıralamıştır. Çoğu KOBİ, küçük işletmelerini nasıl pazarlayacaklarını bilememekte ve pazarlamanın ezici gücü altında zorluk yaşamaktadır. KOBİ’ler pazarlama Çoğu KOBİ, küçük işletmelerini nasıl pazarlayacaklarını bilememekte ve pazarlamanın ezici gücü altında zorluk yaşamaktadır. 2. Kobiler Açısından Pazarlama ve Sorunları KOBİ’ler pazarlama Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 280 kararlarının çoğunu kendi başlarına alırlar ve mevcut fırsatlara ve koşullara çok az pazarlama bilgisi ile yanıt verirler (Scheers, 2018). Son yıllarda yapılan araştırmalar KOBİ’lerin, pazar bölümlendirme, ihtiyaç analizi yapma, ürün ve hizmetlerin başarılı bir şekilde pazarlanması ve pazarlama karmasının uygulanması gibi pazarlama zorluklarını yönetmekte güçlük çektiklerini göstermektedir (Hogarth-Scott vd.,2006; Bowler, Dawood & Page, 2010; Rahman vd., 2016; Scheers, 2018; Şahin & Özüdoğru, 2019). KOBİ pazarlaması, gelişigüzel ve gayri resmi olarak uygulanan pazarlama bilgisi eksikliğinden kaynaklanmaktadır. KOBİ sahiplerinin, işletmelerinin başarısından veya başarısızlığından sorumlu olan belirli kişilik özellikleri sergilediklerinden dolayı, yetersiz pazarlama bilgi ve becerileri, KOBİ’lerde pazarlama sorunları yaratmaktadır (Murphy, 2006). Sorunların çözümü ise KOBİ’lerin öğrenme kapasiteleri, eğitim fırsatlarının mevcudiyeti ve öğrenmeye istekli ve hevesli olmaları ile doğrudan ilişkili olacaktır (Carson & Grant, 1997). 3. Kobiler Açısından Finansal Sorunlar Genel olarak aile şirketi ya da yakın çevre ile oluşturulan ortaklık yapısı ile kurulan KOBİ’ler hayatlarını kurabilmek ve sürdürebilmek için güçlü bir finansal yapıya sahip olmalıdır (Kutlu ve Demirci, 2007: 190). Yalnız kurulurken alabildikleri teşvikler ya da hibe programları ile eksik özkaynak ve yetersiz işletme sermayesi ile kurulabilmektedirler. Bu yapı aynı zamanda kendi bünyesindeki organizasyon yetersizliği yanı sıra ekonominin durumu, siyasi istikrarsızlık, ulusal veya uluslararası pazarda yer alabilme, döviz kuru oynaklığı, yüksek maliyetli krediler, yüksek tutarlı teminatlar, finansal kurumların yaklaşımı ve vergi uygulamaları gibi birçok sıkıntılı durumla da karşı karşıyadır (Yörük, 2001:188; Torlak ve Uçkun, 2005:200). Bu durumlarla baş edebilmeleri için KOBİ’lerin yararlanabileceği, T. C. Ticaret Bakanlığı, Kredi Garanti Fonu (KGF), Kalkınma Ajansları, Türkiye Küçük ve Orta Ölçekli İşletmeler Serbest Meslek Mensupları ve Yöneticiler Vakfı (TOSYÖV), KOSGEB ve KOSGEB’e bağlı hizmet merkezleri bulunmaktadır. Bu kuruluşların teşvik, hibe, proje ya da kredi imkanlarının yanı sıra girişimcilik, finansal danışmanlık, hukuki hizmetler, eğitim, sanayi ile ilişkiler, bilgi ve teknolojiye ulaşım konusunda hizmetleri bulunmaktadır. Fakat KOBİ’lerin ticari hayatın hızlı döngüsü içerisinde zaman kısıtları sebebiyle bu hizmetlerden yararlanabilme olanakları sınırlı olabilmektedir. KOBİ’lerin aile şirketi ya da yakın ilişki içerisindeki ortaklık ile kurulmaları, özellikle finansal açıdan kazanılan gelirin paylaşımı ve işletmede yapılan giderlerin ise özel kişisel harcamalarla karıştırılması gibi problemlerle 281 Serhat Ata / Ali Özer / Nevin Özer karşılaşmalarına neden olmaktadır. Bu iki durumun birbirine karışması, yönetilmesi gereken önemli bir sorun olarak ortaya çıkmaktadır. KOBİ’ler yeterli bir örgüt yapısını zamanla oluşturabilmektedir. İhtiyaç oluştuğunda örgütün içerisine departmanlaşmayı yerleştirerek zamanla kurumsal yapısı oluşmaktadır. Bu sebeple bu durum sancılı bir süreç almakta, yeterli ya da zamanında hareket edebilecek bir örgüt kültürü yavaş ilerleme göstermektedir. Özellikle yakın ilişkili kişiler ile ortaklık yapısından oluşan KOBİ’ler, söz sahibi olan yönetici veya yönetim içerisinde bulunan kişilerin varlığı ile profesyonel yönetilememektedir. Gerekli teknoloji, eğitimli ve tecrübeli personel KOBİ’lerin örgütsel yapısı için oldukça önemli bir rol oynamaktadır. Departmanlaşma yerine ek maliyet yaratan hizmet alımı (reklam, nakliye, muhasebe, finansal hizmetler vb.) ile ve sınırlı sayıda personel üzerinden işleri ilerletme yolunda olmaktadır. Bu ek maliyet ise karlılığı azaltan önemli bir faktördür. KOBİ’lerin en önemli finansal sorunlarının başında yöneticilerin finansal okuryazarlık seviyesi ve organizasyonun içerisindeki finansal yönetimin profesyonelce yapılamamasıdır. Finansal okuryazarlık hem nakit, alacak ve borç yönetimi, hem ihtiyatlı nasıl davranabileceği hem de farklı fon arayışlarının yönetimi açısından önemlidir. Finansal okuryazarlık seviyesinin yüksekliği işletmeye kriz anlarında da stratejik davranabilmeyi sağlayabilmektedir. 4.1. Araştırmanın Amacı Bu çalışmada, Düzce’de faaliyet gösteren KOBİ’lerin finans ve pazarlama alanındaki genel durumunun ortaya çıkarılması amaçlanmıştır. Burada KOBİ’ler incelenirken sektörlerin farklı özellikleri olacağından dolayı Düzce Ticaret ve Sanayi Odası bünyesinden sektörleri temsil etmek için oluşturulan sektör komiteleri hedef grup olarak seçilmiştir. Böylece Düzce ilinde faaliyet gösteren KOBİ’lerin güncel sektöre dayalı pazarlama ve finansal durumu ortaya konarak karşılaşılan sorunlar derinlemesine irdelenecektir. 3. Kobiler Açısından Finansal Sorunlar Ayrıca yüksek finansal okuryazarlık seviyesi ile işletmenin finansal yapısı hakkında mevcut potansiyelini tespit etmesini ve ticari yaşamı süresince de denetimli ve ihtiyatlı hareket edebilmesiyle geleceğe doğru adımlar atabilmesini de sağlayacaktır. Yöneticilerin tüm iş ve işlemleri yürüten stratejik kişi olması, görevlerin paylaşılamaması finansal yönetimle bizzat ilgilenememe ya da yetersiz ilgilenme sorununu ortaya çıkarmaktadır. Finansal durumun net olarak belirlenememesi, nakit politikasının yürütülmemesi ya da ihmal edilmesi, etkin bir alacak tahsilat politikasının yürütülmemesi, finansal yapının riskli durumda olması gibi birçok neden finansal yönetimde başarısızlığa neden olabilmektedir (Tunç ve Köroğlu, 2019:14-15). KOBİ’ler genellikle emek yoğun yapıları sayesinde yeni şartlara, değişen taleplere, teknolojik ilerlemelere hızlı ayak uydurabilmektedir. Tüketicilerin değişen tercihlerine hızlıca cevap verebilen KOBİ’ler oldukça esnek yapıda olabilmektedirler. KOBİ’lerin aldıkları destekler ve teşviklerle yenilikçi ve girişimci yapılarının olması, yapıları küçük de olsa sayılarının çokluğu istihdama katkıları sebebiyle ekonomi içerisinde oldukça önemli bir paya sahiptir (Bayülken ve Kütükoğlu, 2012: 15). Girişimciliğin sembolü olan KOBİ’ler yeni istihdam yaratmada çok etkindir. Yalnız KOBİ’ler enflasyonist Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 282 dönemlerde artan ücret maliyetlerinden dolayı krizi hızlı bir şekilde hissedebilmekte ve olumsuz etkilenmektedir. Enflasyonist dönemlerde yaşadıkları istihdam maliyetinin yanında en önemli sorunlardan biri de artan üretim maliyetleridir. Yüksek faiz, oynak döviz kuru hammadde maliyetlerini de artırmaktadır. Artan maliyetler sebebiyle kısıtlı nakit içerisinde hayatını sürdürmeye çalışan KOBİ’leri oldukça zorlanmaktadır. Yalnız ekonomide yaşanan belirsizlik, enflasyonla birlikte para dolaşımının azalması, ticari bankaların ve yatırım bankalarının sağladıkları krediler içinde KOBİ’lerin kullanabildikleri payların düşük olması, döviz kurunun oynaklığı birçok KOBİ’yi olumsuz etkilemektedir. KOBİ’lerin yetersiz özkaynak ve işletme sermayesi ile kurulmaları işletmelerin risk seviyelerini artırmaktadır. Bu sebeple finansal kuruluşlarından fon sağlama yolunu seçtiklerinde kendilerine zaten az limitlerle ayrılan kredilerden oldukça yüksek maliyetlerle karşılaşma, istenilenin dışında kısıtlı fon alabilme, fon temininde yüksek teminat ve ipotek sorunlarıyla karşılaşma, alınabilecek kredinin vade süresinin kısıtlı olması gibi sorunlarla karşı karşıya bırakmaktadır (Yörük, 2001:190). Ticari bankaların dışındaki finansal kurumların ise yüksek hizmet maliyetlerinin olması KOBİ’ler tarafından tercih edilememesine sebep olmaktadır. 5.1. KOBİ’lerin Pazarlama ile İlgili Yaşanan Sorunların Bulguları Bu bölümde olgubilim (fenomenoloji) deseni kullanılarak elde edilen verilerin analizi sonucunda 71 kod elde edilmiştir. İlgili kodlardan benzer özelliklere sahip olanlar toplanıp 12 kategoride gruplandırılmıştır. Analizin son aşamasında bir araya getirildiğinde anlamlı bir bütün oluşturan kategoriler temalara göre gruplandırılarak 7 temel tema elde edilmektedir. Toplanan verilerin analizi sonucunda ulaşılan kategori ve temalar Tablo 6’de sunulmaktadır Tablo 6. Pazarlama İle İlgili Kategoriler ve Temalar Pazarlamada Tecrübe, Eğitim ve Bilgi Eksikliği Satış Personeli Becerileri Fiyat İstikrarsızlıkları ve Belirsizlikler Ekonomik Değişim Haksız Rekabet Yeni Pazar Fırsatları ve Rol Model Rekabet Stratejileri Ürün Bilgisi ve Ambalajlama Fiyat İstikrarsızlıkları Dijitalleşme ve Reklam Lojistik Maliyetler Pazarlama Karması Sorunları Tam Zamanında Teslim Personel-Müşteri İlişkileri Müşteri İlişkileri Yönetimi Coğrafi ve Lojistik Konum Konumlandırma Stratejisi Yeniliklerin Takibi Dijitalleşme Sorunları Tablo 6. Pazarlama İle İlgili Kategoriler ve Temalar 4.2. Araştırmanın Yöntemi Bu çalışmada, verilerin toplanmasında nitel bir yöntem olan yarı yapılandırılmış mülakat tekniği kullanılmıştır. Araştırma deseni olarak da günlük hayatta karşılaşılan olay, deneyim, algı ya da durumlar gibi yabancı olmadığımız veya anlamını tam anlamıyla bilmediğimiz farklı olguları araştırmayı amaçlayan (Yıldırım ve Şimşek, 2016, s. 69) olgubilim (fenomoloji) kullanılmıştır. Düzce’deki Kobilerin genel finans ve pazarlama durumu ortaya çıkarılırken sektör temsilcilerine yöneltilen sorularda alanında uzman akademisyenlerin görüşleri de dikkate alınarak oluşturulmuştur. 283 Serhat Ata / Ali Özer / Nevin Özer Komitelerle yüz yüze toplantılar yapılarak, mülakat tekniği ile bilgiler toplanmıştır. Her bir sektör temsilcisi ile farklı günlerde yaklaşık iki saatlik bir görüşme yapılmıştır ve katılımcılardan müsaade alınarak ses kayıt cihazı ile görüşmeler kayıt altına alınmıştır. Çalışmanın güvenilirlik ve geçerliliği sağlamak adına katılımcıların ifadelere verdikleri cevaplar sonuç olarak literatürdeki benzer çalışmaların sonuçları ile mukayese edilerek tutarlılıkları güvenilirlik için saptanmıştır. Geçerlilik adına ise her sektörden farklı temsilcilere benzer sorular yöneltilerek örneklem çeşitlendirilmiştir. Komitelerle yüz yüze toplantılar yapılarak, mülakat tekniği ile bilgiler toplanmıştır. Her bir sektör temsilcisi ile farklı günlerde yaklaşık iki saatlik bir görüşme yapılmıştır ve katılımcılardan müsaade alınarak ses kayıt cihazı ile görüşmeler kayıt altına alınmıştır. Çalışmanın güvenilirlik ve geçerliliği sağlamak adına katılımcıların ifadelere verdikleri cevaplar sonuç olarak literatürdeki benzer çalışmaların sonuçları ile mukayese edilerek tutarlılıkları güvenilirlik için saptanmıştır. Geçerlilik adına ise her sektörden farklı temsilcilere benzer sorular yöneltilerek örneklem çeşitlendirilmiştir. 5. Bulgular 5.1.1. Satış Personeli Becerileri Bu tema tecrübe, eğitim ve beceri eksikliği ve pazarlama bilgisi eksikliği kategorilerinin bir araya gelmesi ile meydana gelmiştir. Ayrıca KOBİ’lerdeki satış personellerinin temel becerilerini içeren nitelikli pazarlama eğitimi, tecrübe ve satış tecrübesi kazanma gibi kodları barındırmaktadır. 284 \| Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 5.1.1.1. Pazarlamada Tecrübe, Eğitim ve Bilgi Eksikliği KOBİ’lerde hem işletme sahibinin hem de çalışanların pazarlama konusunda tecrübe eksikliği en önemli sorunlardan bir tanesidir. Genelde işletme yönetiminin pazarlama konusunda uzmanlığı, deneyimi ve eğitimi yetersizdir ve tüm işletme operasyonları bir veya küçük bir grup birey tarafından gerçekleştirilmektedir. Bu durum, işletmenin pazarlama fonksiyonunu yerine getirmesini engellemektedir. Özellikle şirket yönetimi, organizasyon içindeki her işlevi tek bir çalışanla yerine getirmeye çalışırken, özellikle büyük firmaların değişen pazar koşullarına uyum sağlamakta zorlanmaktadır. Çeşitli nedenlerden dolayı, KOBİ’ lerin yöneticiler de dâhil olmak üzere tüm çalışanlara dışarıdan eğitim sağlama olasılığı büyük firmalara göre daha düşüktür. Sınırlı kaynaklar ve bilgi eksiklikleri küçük firmaların yönetim eğitiminden elde edecekleri faydaların daha az farkında olmalarına neden olmakta ve çok azı eğitimi stratejik bir araç olarak görmektedir. Çünkü pazarlamaya yönelik eğitim, tecrübe ve bilgi eksikliği sorunları, “KOBİ’lerin pazarlamayı ‘büyük firmaların yaptığı bir şey’ olarak küçümsemesinden kaynaklanmaktadır. Bu bakış açısı, KOBİ’lerin üretime dayalı yapılarının ve pazarlama konusundaki deneyim eksikliklerinin bir sonucudur. Ayrıca, birçok KOBİ’de iyi tanımlanmış bir kariyer yolunun ve yapılandırılmış eğitim programlarının bulunmaması, uzmanlık becerileri geliştirmek isteyen çalışanlar için bir zorluk teşkil etmektedir. Araştırmaya katılan komite üyelerinin KOBİ’lerde pazarlama faaliyetleri için nitelikli personele ihtiyaç duyulup duyulmadığına dair soruya verdikleri cevaplar incelendiğinde katılımcıların tamamı nitelikli pazarlama eğitimi, tecrübesi ve becerisi olan personele ihtiyaç duyduğu yönünde cevap vermişlerdir. Komiteler pazarlama konusunda bilgisiz ve eğitimsiz personelin müşteriyi ve satışları olumsuz etkilediğinden bahsetmişlerdir. Bir katılımcı bu eksen kaymasını şu şekilde ifade etmiştir: “Çalışanlar, nasıl üretilir konusunda iyi olsa da nasıl pazarlanır’ı bilmiyorlar. Ayrıca nasıl pazarlanır’ı öğrenmeye başlayan da büyük firmalara kayıyor çünkü nitelikli personelin risk algısı çok düşük”. 5.1.2.1. Fiyat İstikrarsızlıkları 5.1.2.1. Fiyat İstikrarsızlıkları Ekonomik koşullar genellikle KOBİ’ler için pazarlama stratejilerinde yenilik ve uyarlama gerektirmektedir. Fiyatların dinamik olduğu ekonomik belirsizlik dönemlerinde tüketici davranışları ve tercihleri değişebilmektedir. Fiyat istikrarsızlığı ve enflasyonun yüksek olduğu dönemlerde KOBİ’lerin hedef pazarlarını yeniden değerlendirmeleri ve mesajlarını buna göre uyarlamalarını gerektirmektedir. KOBİ’ler sınırlı kaynaklarla, kendilerini enflasyonla ilişkili maliyetleri karşılamakta zorlanırken bulabilmekte ve bu durum potansiyel olarak fiyat artışları ve müşteri kaybına yol açabilmektedir. Hükümet müdahaleleri, politikaları ve destek programları KOBİ’lerin pazarlama faaliyetlerini önemli ölçüde etkileyebilmektedir. Ekonomik belirsizlik dönemlerinde hükümetler işletmeleri desteklemek için teşvik paketleri veya teşvikler sunabilmektedir. Bu konuda yaşanacak aksaklıkların işletmelerin sadece pazarlama faaliyetlerini değil tüm politikalarını etkilemesi kaçınılmaz hale gelebilmektedir. Nitekim bir katılımcının vermiş olduğu cevabın söz konusu yargıyı destekler nitelikte olduğu görülmektedir: “Biz ancak alabilirsek satarız. Pandemi ve sonrası dönemde yaşanan ekonomik istikrasızlıklarla birlikte sabit müşterilerimiz azalmıştır. Hammadde tedarikinde yaşanan fiyat istikrarsızlıkları, devlet eliyle sabit fiyat uygulamasının olmaması, ihracata konan kotalar tüm politikamızı etkilemiştir. Ayrıca bu dönemde iç piyasada mal fazlalığı meydana gelmiş böylece birbirine bağlı sektörler adeta domino etkisi yaratmıştır. Devlet tarafından bizlere uygulanan KDV’ de yeterli düşüşün olmayışı tüm alım gücümüzü etkilemiştir” Araştırmaya katılan komitelerin büyük çoğunluğu ekonomik koşulların pazarlamaya etkilerini fiyat istikrarsızlığı ekseninde açıklamaya çalışmışlardır. Ayrıca fiyatlarda yaşanan dalgalanmaların neticesinde oluşan belirsizlik ve risk algısı komitelerce tartışılan bir diğer önemli husus olarak belirtilmiştir. Ayrıca fiyat dalgalanmalarının hammadde fiyatlarını etkilediğini ve dolayısıyla azalan üretim ile birlikte müşteri talep ve beklentilerine verilecek cevapların geciktiğini vurgulamışlardır. 5.1.2. Ekonomik Değişim Bu tema altında ekonomi temelli değişimleri içeren fiyat İstikrarsızlıklarının ve belirsizliklerin pazarlama faaliyetlerini nasıl şekillendirdiğini içeren kodlar bulunmaktadır. 285 Serhat Ata / Ali Özer / Nevin Özer \| 5.1.3.1. Haksız Rekabet KOBİ’ler büyük şirketlerle rekabet ederken sayısız zorlukla karşılaşmaktadır. En önemli engellerden birisi sınırlı finansal kaynaklardır. Büyük muadillerinin aksine, KOBİ’ler genellikle sermaye açısından kısıtlamalarla karşı karşıya kalmakta, bu da araştırma ve geliştirme, pazarlama ve teknoloji altyapısına yatırım yapma kabiliyetlerini etkilemektedir. Büyük şirketler toplu satın alma, üretim verimliliği ve kapsamlı dağıtım ağlarından faydalanarak KOBİ’lerin yakalamakta zorlandığı bir maliyet avantajı elde etmektedir. Katılımcılar Düzce ilinde faaliyet gösteren büyük firmaları varlığının pazarlama açısından değerlendirdiklerinde komitelerin bir kısmının söz konusu varlığın standardizasyon sağlayarak rekabet ve kaliteyi artıracağı, bir kısmının ise haksız rekabete yol açarak markalaşma ile birlikte hem nitelikli personeli hem de müşteriyi etkileyerek küçük firmaların faaliyetlerini olumsuz yönde etkileyeceği doğrultusunda cevaplar vermişlerdir. Büyük firmaların uyguladıkları promosyonlar, büyük rakiplerin yeni şube açmaları, haksız rekabet, teknoloji eksikliği, dijitalleşmeye dayalı markalaşma, nitelikli çalışanların tercih yönünün değişmesi, rekabet gücünün daralması ve ürün farklılaşmasına gidememe KOBİ’lerin büyük firmalara karşı eksik yönlerini oluşturmaktadır. Bu doğrultuda bir katılımcı rekabet kaynaklı olumsuzlukları şu şekilde ifade etmiştir: “Büyük firmaların varlığı dijitalleşme çağında haksız rekabete yol açmaktadır. Nitekim büyük markalar bu şehre geldiğinde bizler arasında rekabet yoğunlaşmakta iken büyük firma aradan sıyrılmaktadır. Az sayıdaki personelimiz de büyük firmalara kaçabilmektedir.” 5.1.3. Rekabet Stratejileri Bu tema haksız rekabet, yeni pazar arayışı ve rol model oluşumu kategorilerinin bir araya gelmesi ile meydana gelmiştir. Bu doğrultuda yeni şubeler açma, pazar derinliğine inme, farklılaşma stratejisi gibi kodları da içermektedir. 286 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 5.1.3.2. Yeni pazar fırsatları ve Rol Model Oluşumu Büyük şirketler en son teknolojilere yatırım yapacak finansal güce sahip olabilirken, KOBİ’ler dijital platformlardan yararlanarak küresel bir kitleye çok daha az maliyetle ulaşabilmektedir. Çünkü doymuş pazarlarda yerleşik markalarla doğrudan rekabet etmeye çalışmak yerine, KOBİ’ler genellikle niş pazarları hedefleyerek başarıya ulaşabilmektedir. Niş pazarlar, daha büyük rakipler tarafından göz ardı edilebilecek belirli tüketici ihtiyaçlarına veya tercihlerine hitap etmelerine olanak tanır. Bunu yaparken, KOBİ’ler kendilerini seçtikleri alanda uzman olarak konumlandırabilir, tam olarak uyarlanmış ürün veya hizmetler arayan tüketiciler arasında güvenilirlik ve güven oluşturabilirler. 287 Serhat Ata / Ali Özer / Nevin Özer E-ticaret, sosyal medya ve dijital pazarlama da KOBİ’lere pazarlara ve tüketicilere eşi benzeri görülmemiş bir erişim sağlayarak oyun alanını bir ölçüde eşitlemiştir. Bunun yanında, yeni pazarlarda büyük firmaların getirdikleri yenilikler ile iş birliğine ve kaliteye teşvik etmesi, rekabet gücü yaratması da artı yönlerini oluşturmaktadır. Bu doğrultudaki bir katılımcı görüşü aşağıdaki gibidir: “Büyük firmaların varlığı aslında bize standardizasyon sağladı. Bizlerin gelişiminde rol oynadılar ve yeni pazarlar keşfetmemizi sağladılar.“ Ayrıca, değişen tüketici tercihleri ve sürdürülebilirlik ile yerelliğe verilen önemin artması, KOBİ’lerin kendilerini farklılaştırmaları için fırsatlar yaratmıştır. Yerel işletmeleri destekleme söylemi ve benzersiz, zanaatkar ürünlere olan talep, tüketici davranışını küçük işletmeler lehine değiştirebilmektedir. 5.1.4. Pazarlamanın Karması Sorunları Bu tema ürün bilgisi ve ambalajlama, fiyat istikrarsızlıkları, dijitalleşme ve reklam ile lojistik maliyetler kategorilerinden oluşmaktadır. Pazarlamanın 4P’sine dair karşılaşılan sorunların temelinde yaşanan fiyat dalgalanmaları yatmaktadır. Çoğunlukla hammaddeye dayalı üretim hattında yaşanan temin sorunları ile birlikte artan stok bulundurma maliyetleri, ürünlerin paketlenmesi ve ambalaj malzeme fiyatlarının artması belirlenen kodlar arasındadır. 5.1.4.3. Dijitalleşme ve Reklam Gerekli becerilere sahip olmayan KOBİ’ler dijital kampanyalarını etkili bir şekilde optimize etmekte ve yatırım getirilerini ölçmekte zorlanabilir. KOBİ’lerin son yıllarda karşılaştığı dijitalleşme ve reklam sorunlarının ucu yine fiyat istikrarsızlığına dayanmaktadır. Tutundurma yönünden reklam bütçelerinin düşmesi fuar masraflarının artması, yerel reklam konusundaki eksikliklerle birlikte sosyal medya kanallarının kullanımında bilgi ve takibe dayalı eksiklikler KOBİ’lerin temel sorunları olarak dikkat çekmektedir. Benzer doğrultuda bir katılımcının görüşü aşağıdaki gibidir: “ Fuarlarda tanıtım maliyetleri artık bizlerin bütçesini aşmıştır. Dahası itibar katacak yerel reklamlara dahi bütçe ayırmakta zorlanmaktayız. Dijital çağda en büyük çıkış noktamız sosyal medya kanallarında tanıtım. Fakat onda da teknik eksiklikler mevcut. Markalaşamamaya dayalı tanıtım eksikliklerimiz var.” 5.1.4.1.Ürün Bilgisi ve Ambalajlama Eksik ürün bilgisi ve kötü tasarlanmış ambalajlar tüketicilerin ilgisini çekmede, doğru mesajı iletmede ve markanın değerini müşteriye aktarmada önemli bir sorundur. KOBİ’ler fiyat artışlarının olduğu dönemlerde bütçe kısıtlamalarıyla karşı karşıya kaldığı için ambalaj malzemelerinin kalitesinden ve tasarımından ödün verebilmektedir. Bu sebeple ürünle veya hedef kitleyle uyumlu olmayan ambalajlar müşteri veya pazar fırsatların kaçırılmasına yol açabilmektedir. Ayıca, KOBİ’ler, özellikleri, faydaları ve benzersiz satış noktaları da dâhil olmak üzere kendi ürünleri hakkında kısıtlı bilgiye sahip olabildiği için çalışanların ürün detaylarına ilişkin yetersiz eğitimi, müşterilere yanlış veya eksik bilgi verilmesine neden olabilmektedir. KOBİ’ler rekabet veya ekonomik koşullar kaynaklı pazar trendleri, müşteri tercihleri ve rakip ürünler hakkında bilgi sahibi olmakta zorlanabilmektedir. Kötü hazırlanmış pazarlama materyalleri ve net olmayan mesajlar(hem çevrimiçi hem de çevrimdışı çeşitli platformlarda ürün bilgilerindeki tutarsızlık) müşterilerin kafasının karışmasına neden olabildiği için karşılaşılan sorunlar arasında Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 288 yerini almaktadır. Nitekim bir katılımcı söz konusu sorunları şu şekilde ifade etmiştir: “Ambalaj malzeme fiyatları son yıllarda muazzam arttı. Bu durum da stokçuluğu beraberinde getirdi. Bir de üstüne paket servis yapacak elemanların niteliksizliği eklenince birçok sorun ile karşı karşıya kaldık. Müşteriye beğendirebilmek için kendi ambalajımızı kendi paketimizi kendimiz üretir hale geldik. Böylece markalaşamama son fiyatlara yansıdı.“ 5.1.4.2. Fiyat İstikrarsızlıkları Fiyat ise başlı başına bütün sorunların temelini oluşturduğundan tüm pazarlama karması faktörlerine yansımaktadır. Vadeli alım satımlar müşteri tercihlerini etkilemektedir. Firmalar ihale usulü alınan ürünlerdeki fiyat farklılıklarından ciddi olarak etkilenmektedir ve bu durum ürün fiyatlarına da yansıyabilmektedir. Bir katılımcının verdiği görüşler de benzer şekilde aşağıdaki gibidir: “Fiyatlardaki istikrarsızlık, firmalar hammaddeden son tüketiciye kadar birbirine bağımlı olduğu için maliyetleri ciddi olarak arttırmıştır. “ 5.1.4.4.Lojistik Maliyetler KOBİ’ler genellikle sınırlı bir tedarikçi ve dağıtım ağına bağlıdır. Özellikle akaryakıt fiyatlarının arttığı dönemlerde tedarik zincirinde lojistik sorunlar, jeopolitik olaylar veya diğer dış faktörlerden kaynaklanan herhangi bir aksama, ürünün bulunabilirliğini ve teslimatını önemli ölçüde etkileyebilmektedir. Bu tür kırılganlıklar KOBİ’lerin itibarına zarar verebilmekte ve müşteri güveninin olumsuz yönde etkilenmesine neden olabilmektedir. 289 Serhat Ata / Ali Özer / Nevin Özer “Lojistik maliyetler ve hammaddeye ulaşım sıkıntısı firmamızın başlıca sorunlarındandır. Akaryakıt fiyatlarındaki artış ciddi manada belimizi büktü. 5.1.5. Müşteri İlişkileri Yönetimi Bu temada tam zamanında teslim ve personel-müşteri ilişkileri yönetimi gibi kategoriler bulunmaktadır. Söz konusu kategoriler aşağıdaki gibi sınıflandırılarak açıklanmıştır. 5.1.5.1. Tam Zamanında Teslim Üretim süreci aşamasında siparişe dayalı üretim yapan KOBİ’lerin malzeme ihtiyacı olduğunda düşük hacimli siparişlerden kaynaklı iskonto sorunları oluşabilmektedir. Böyle bir durumda birim maliyetler yeterince düşürülememekte ve söz konusu durum fiyatlara yansıyabilmektedir. Siparişe dayalı üretim kaynaklı tam zamanında teslim KOBİ’ler için en önemli hususlardan biridir. Üretim için tam zamanında tedarik ayrıca ürünün müşteriye tam zamanında teslim edilebilmesinin ön koşuludur. Bir katılımcının görüşü şu şekildedir: “Ürünün ham maddesini vadeli olarak alamadığımız şu dönemde teslimat kaynaklı sorunlar ve şikâyetlerin sayısı oldukça arttı. “ 5.1.5.2. Personel-Müşteri İlişkileri Büyük firmalara göre daha esnek yapıda örgütlenen KOBİ’lerde müşteriler ile birebir temas kurabilme olasılığı olduğu görülür. Çünkü KOBİ’lerin doğrudan satış yapabilme özelliği olduğu gibi müşteri sorunlarına da birebir temas sayesinde doğrudan erişebilme olanağı bulunmaktadır. Ayrıca müşteri merkezlilik politikası güden firmalar müşteri özelinde ürün üretebilme gibi seçenekleri sunmada öncül olabilmektedir. Fakat tüm ihtimaller tamamen nitelikli personelin varlığına bağlıdır. Nitekim bir katılımcının görüşü şu şekildedir: “Kalifiye eleman eksikliği müşteri şikâyetlerinin artmasına yol açtı. Çünkü elemanlar müşterileri yeterince dinlemiyor ve dikkate almıyorlar. Bu yıl bu yüzden çok fazla müşteri kaybettik.” 5.1.6. Konumlandırma Stratejisi Bu bölümde konumlandırma teması coğrafi ve lojistik açıdan ele alınarak tek bir kategoride toplanmıştır. 290 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 5.1.6.1. Coğrafi ve Lojistik Konum Düzce konum olarak genel anlamda coğrafi açıdan büyük şehirlere yakınlığı sebebi ile bir sorun olmaktan ziyade avantaj oluşturmaktadır İstanbul ve Ankara’ya yakınlık sebebiyle hammadde ve işgücüne kolay ulaşılabilirliği, pazarlara yakın olması ve lojistik ağı üzerinde olması katılımcılarca büyük etki olarak görülmektedir. Bunula birlikte, lojistik açıdan konum itibari ile büyük firmaları cezbetmesi de rekabet açısından KOBİ’leri zorlayıcı unsur olarak değerlendirilebilir. KOBİ’ler ayrıca bulundukları coğrafyada sağladıkları istihdam ile büyük şehirlere göçü engellemektedir. Dolayısıyla bölgesel gelişmişlik düzeylerindeki farklılıkların önünde bir engel olarak durmaktadırlar. Ayrıca sağladıkları ekonomik döngü ile bölgelerindeki girişimcilik potansiyelini körükleyerek yeni iş alanlarının açılmasının önünü açmaktadırlar. Benzer şekilde katılımcıların görüşü şu şekildedir: “Düzce coğrafi açıdan büyük avantaj sağlamaktadır. İşgücü potansiyeli yüksektir. İstanbul ve Ankara’ya yakınlık rekabet avantajı sağlamaktadır. Hammadeye kolay erişilebilir, büyük limanlara yakındır fakat şehre de bir liman ve gümrük ağı gerekmektedir.” 5.1.7. Dijitalleşme Sorunları Dijitalleşme sorunları teması altında yeniliklerin takibi kategorisi oluşturulmuştur. 5.1.7.1. Yeniliklerin Takibi Günümüz rekabet ortamının en önemli özelliği, benzersiz ve müşteriye özel ürünler üretebilme kapasitesidir. Tüm kuruluşlar gibi KOBİ’ler de küresel pazardaki değişikliklere yanıt olarak maliyet, kalite, hız ve hizmet üstünlüğü peşinde koşmak zorundadır. Ayrıca sürekli ve hızlı yenilik kapasitesine de sahip olmaları gerekmektedir. Dolayısıyla KOBİ’lerin rekabet stratejilerinin temelinde yenilik yatmaktadır. Müşterilerin mevcut ihtiyaçlarını karşılamaktan ziyade, onların ihtiyaçlarını öngörmek ve onlara geniş bir mal ve hizmet yelpazesi sunmak şeklinde bir değişim meydana gelmekte, bu da işletmelerin üretim ve satış sistemlerinde farklılaşmaya yol açmaktadır. Esnek üretim ve yeni tasarıma dayalı rekabetçi ortamlarda zamanında üretim, yüksek kaliteli ürünler ve yenilikçi tasarımın önemi artmaktadır. Söz konusu yenilikleri yapabilmek için de gerekli donanıma ve teknolojiye sahip olmak gerekir. Bir katılımcı görüşü şu şekildedir: “Büyük firmalar teknolojiyi en etkin biçimde kullanırken bizim yeterli düzeyde donanımımız yok. Onlar yapay zekâ kullanırken bizim tek kaynağımız sosyal medya.” 291 Serhat Ata / Ali Özer / Nevin Özer \| 2 5.2. KOBİ’lerin Finansman İle İlgili Yaşadıkları Sorunların Bulguları Bu bölümde olgubilim (fenomenoloji) deseni kullanılarak elde edilen verilerin analizi sonucunda 68 kod elde edilmiştir. İlgili kodlardan benzer özelliklere sahip olanlar toplanıp 17 kategoride gruplandırılmıştır. Analizin son aşamasında bir araya getirildiğinde anlamlı bir bütün oluşturan kategoriler temalara göre gruplandırılarak 6 temel tema elde edilmektedir. Toplanan verilerin analizi sonucunda ulaşılan kategori ve temalar Tablo 6’da sunulmaktadır Tablo 7. Finansman Soruları İle İlgili Kategoriler ve Temalar Yetersiz Özkaynak ve Alacak Yönetimi Organizasyon Yapısı ve Profesyonel Yönetim Finansal Danışmanlık, Finansal Teknik Bilgisi, Finansal Okuryazarlık Seviyesi Finansal Yönetim Enflasyon Maliyet yüksekliği Ekonomi Kredi Yüksek Teminat ve İpotek Banka Destek Kuruluş Kredileri Sektörel Finansman Desteği Danışmanlık Hizmetleri Destek Kuruluşlar Finansman Teşviği Enerji Teşviği Vergi Teşviği Yatırım Teşviği Teşvik Eğitim Kuvvetli İletişim İşbirliği Beklentiler Tablo 7. Finansman Soruları İle İlgili Kategoriler ve Temalar 5.2.1. Finansal Yönetim Bu tema tecrübe, yetersiz özkaynak ve alacak yönetimi, organizasyon yapısı ve profesyonel yönetim, finansal danışmanlık, finansal teknik bilgisi, finansal okuryazarlık seviyesi kategorilerinin bir araya gelmesi ile meydana gelmiştir. 292 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 5.2.1.1. Yetersiz Özkaynak ve Alacak Yönetimi KOBİ’lerin kurulurken eksik özkaynak ile kurulmaları belki de yaşadıkları sorunların en başında gelmektedir. Küçük tasarruflarla ya da hibe destekleri ile kurulan işletmeler bu finansmanı, üretim teknolojilerinde, pazara yerleşmede, üretimlerinde, işletmeyi ilerletmelerinde, nakit-alacak-borç yönetiminde de kullanmaktadırlar. (Uçkun, 2009:123). Bu sebeple zaten yetersiz başlayan işletme, ticari hayatı boyunca da bu zorlukla hep karşı karşıya gelmektedir. Araştırmaya katılan komite üyelerinin sektördeki firmaların yaşadığı finansal sorunlar sorusuna verdiği ilk cevabı özkaynak yetersizliği oluşturmaktadır. Gelişmekte olan ülkelerde yaşanan en büyük sorunlardan biri dolaşımdaki paranın az olması kaynaklı ticari işlemlerin borçlanarak sürdürülmesidir. Bu sebeple işletmeler yetersiz sermaye ile ticari hayatlarına başlamaları ve sürdürmeye çalışmaları, işletmelerin nakit-alacak-borç yönetimini de etkilemektedir. Ticari hayatın içerisinde müşterilere sağlanan alacak politikası işletmenin nakde ulaşma hızını göstermektedir. Nakte ulaşım hızlı ise borç yönetimin de, yatırım yapmakta ayrıca rekabet etmekte daha kolay olabilmektedir. Bu sebeple işletmeler ne kadar etkin alacak tahsilat politikası yürütebilirse borç ödeme politikalarını da başarılı yönetebileceklerdir. Yalnız işletmelerin nakit ve alacak yönetimini profesyonel yönetememeleri işletmeler açısından yaşanan çok önemli bir finansal yönetim sorunu olarak ortaya çıkmaktadır. Araştırmaya katılan komite üyelerinin sektördeki firmaların yaşadığı finansal sorunlar sorusuna verdiği cevaplardan bir diğeri ise alacak yönetiminin profesyonelce yönetilememesi olmaktadır. Bir komite üyesinin bu konuda verdiği cevap; “Alacakların yönetimi çok önemli sorun olmaktadır. Alacakları finanse etmek zorunda kalabiliyorsunuz. Siz müşteriniz yerine ödemeyi yapmaktasınız yalnız müşteriden daha sonra bu ödemeyi almakta zorluk yaşamaktasınız, alamadığınızda ve bu durumu sürekli yaşadığınızda iflasa kadar sürükleyebilir. Tahsilatını yapmadığınız iş sizin değildir. Bu sebeple alacakların tahsil edilmesi ve yönetilmesi önemli bir zorluktur.” 5.2.1.2.Organizasyon Yapısı ve Profesyonel Yönetim Araştırmaya katılan komite üyelerinin sektördeki firmaların ayrı olarak muhasebe ve finans departmanı olup olmadığı sorusuna verdiği cevaplarda bir sektör hariç diğerlerine ayrıca bir muhasebe ve finans departmanın olmadığı yönündedir. Muhasebe işlemlerini ön muhasebe işlemi ile dışardan muhasebe hizmeti alımıyla yapıldığını belirtmişlerdir. Bir sektörde belirli bir büyüklüğe ulaşan firmalarda departmanlaşmanın olduğunu söylemişlerdir. 293 Serhat Ata / Ali Özer / Nevin Özer İşletmenin yönetiminin ve işletmelerin finansman yönetiminin ise daha çok işletme sahibi tarafından yönetildiği cevabını vermişlerdir. 5.2.1.3. Finansal Danışmanlık, Finansal Teknik Bilgisi ve Finansal Okuryazarlık Seviyesi Araştırmaya katılan komite üyelerinin Bakanlıklar, KOSGEB, Kalkınma Ajansları gibi kurumların verdikleri teşvik ve danışmanlık hizmetlerinin yeterli olup olmadığı sorusuna verdiği cevaplarda 3 sektör danışmanlık hizmetini yeterli bulurken kalan temsilciler verilen danışmanlık hizmetlerini yetersiz bulmaktadır. Firma ziyaretlerinin az olması, KOBİ’lerin yoğun olduğu bölgelerde kurumlarının ofislerinin olması gerektiğini böylelikle ihtiyaç duyulduğunda ya da danışmanlık alınmak istendiğinde ulaşılabilir olmalarını sağlayacağı yönünde cevap vermişlerdir. Ayrıca iki temsilci bu danışmanlık hizmetlerinin alımında firma sahiplerinin de istekli olmalarının gerekliliğini ve zaman ayırabilmeleri gerekliliğini de vurgulamışlardır. Sektör temsilcilerine finansal kuruluşlardan yararlanma sorusuna verdikleri cevaplarda finans kuruluşlarını tanıdıklarını yalnız daha çok Bakanlıklar, KOSGEB, Kalkınma Ajansları vb. kuruluşlar ve Ticari bankaları ve bu kuruluşların ürünlerini kullandıklarını belirtmişlerdir. Ayrıca sektör temsilcilerine atıl durumda kalan nakitlerini nasıl değerlendirdikleri sorusuna ise finansal ürünlere yatırımı zaman zaman yaptıklarını da belirtmişlerdir. Verilen cevaplardan sektör temsilcilerin hem yatırım ürünleri hem finansman kuruluşları hem de nasıl kullanılacağını ne gibi fayda-maliyetle karşılaşacaklarını bilme konusunda finansal okuryazarlık seviyelerinin yüksek olduğu anlaşılmaktadır. Komite temsilcilerine sektördeki firmaların finansal teknikler bilgi durumu ve kullanımı hakkında sorulan soruya verdikleri cevaplar barter, leasing ve ticari banka ürünlerinin daha sık kullanıldığını, factoring, forfaiting gibi yöntemlerin kimi temsilcilerin bildiğini fakat yüksek hizmet maliyetleri sebebiyle kullanmadıklarını, kimi temsilcilerin ise fikirlerinin olmadığını ve bu konuda eğitim verilmesi gerekliliğini, kimi temsilcilerin ise banka danışmanlık işlemlerinde farklı ürünler ve yöntemler konusunda yönlendirilebildikleri cevaplarını vermişlerdir. 5.2.2. Ekonomi Bu tema enflasyon ve maliyet yüksekliği kategorilerinin bir araya gelmesi ile meydana gelmiştir. Türkiye gibi gelişmekte olan ekonomiler ulusal/ uluslararası politika, siyaset, sermaye piyasaları, makroeokonomik birçok faktörden etkilenmektedir. Bu ülkelerin ekonomileri oldukça kırılgan 294 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme olmaktadır. Özellikle yaşanan yüksek enflasyon ise KOBİ’leri hem yükselen maliyetler hem de nakite ulaşım sıkıntısı ile borçların ödenememesi, yüksek maliyetle borçlanma, yüksek faizlerle karşılaşma ya da istenilen finansman desteğine ulaşılmaması gibi birçok sıkıntı ile başbaşa bırakmaktadır. Araştırmaya katılan komite üyelerinin sektördeki firmaların yaşadığı finansal sorunlar sorusuna verdiği cevaplarda çok sık olarak yüksek maliyetler cevabında yoğunlaştığı görülmüştür. Hammadde fiyatlarının yüksek olması, enerji fiyatlarının yüksek olması ayrıca borçlanma maliyetlerinin de yüksek olması işletmeleri bu konularda yaşanılan ciddi sıkıntı olarak görmektedirler. 5.2.3. Banka Bu tema kredi ve yüksek teminat ve ipotek kategorilerinin bir araya gelmesi ile meydana gelmiştir. Ayrıca alt kategoriler içerisinde kredi taksit ödeme zorlukları, yüksek tutarlı kredilere ulaşım sıkıntısı, piyasadan borçlanma, Eximbank kredileri, rotatif kredileri ve becayiş kredileri de oluşturmaktadır. Komite üyelerinin sektördeki firmaların finansal ihtiyaçlarını karşılama kanallarının ne olduğu sorusuna verilen cevaplar özellikle ticari banka kredileri üzerine yoğunlaşmaktadır. Ayrıca KGF kredilerinin de kullanıldığını belirtmişlerdir. Komite üyelerinin sektördeki firmaların yaşadığı finansman sorunlarına verdiği cevaplarda ise Ticari bankaların sağladıkları kredilerde faiz oranlarının yüksekliği, istenilen miktarda kredilere ipotek verilmesine rağmen bankaların bu kredi temininde isteksiz davranmaları, teminatlı kredilerin yetersiz kalması, bankaların yüksek miktardaki kredileri vermek istememeleri ve enflasyonist ortamda kaynak yetersizliği sebebiyle banka kredi taksitlerinin ödenmesinde yaşanılan zorlukları belirtmişlerdir. Komite üyeleri özellikle devlet bankalarının kredi konusunda esnek davranmalarını da talep etmişlerdir. Komite üyelerinin sektördeki firmaların yaşadığı finansman sorunları için öneriler sorusuna verdiği cevaplar arasında kullanılan banka kredileri olsun ya da destek veren kuruşlardan yararlanılan krediler ya da hibeler olsun mutlaka kullanan firmaların sıkı takip edilmesi gerekliliği yönünde de cevaplar vermişlerdir. 5.2.4. Destek Kuruluşlar Bu tema vergi teşviği, destek kuruluş kredileri, sektörel kredi desteği, danışmanlık hizmetleri kategorilerinin bir araya gelmesi ile meydana gelmiştir. Bakanlıklar, KOSGEB, Kalkınma Ajansları gibi kurumlar girişimcilere verdikleri hem danışmanlık hizmetleri hem finansman destekleri 295 Serhat Ata / Ali Özer / Nevin Özer ile şirketlerin kuruluşlarından itibaren ticari hayatlarının yanında olmaktadır. Ticari hayatın zamansal döngüsü içerisinde firma yöneticilerinin de zaman ayırabilmesi, istekli olması ya da ihtiyaç duyduğunda destek almaktan yana olması ile bu kurumların her hizmetin yararlanabilmektedir. Yani firmanın üzerinde bu kurumların verdiği hizmetlerden yararlanabilme konusunda talepkar olması da gerekmektedir. Komite üyelerine bu kurumların sunduğu teşvik ve danışmanlık hizmetinin yeterli olup olmadığı sorusuna kimi sektör temsilcisinin yeterli kimi sektör temsilcisinin ise yetersiz olduğu yönünde cevaplar alınmıştır. Devlet teşvikleri hakkında bilgi sahibi olunması, verilecek finansman destekleri ve hibe programlarından haberdar olunabilmesi bu kuruluşların takip edilebilmesi ve bu kurumlarında firmaları takip edebilmeleri ile gerçekleşebilmektedir. Bu sebeple komite üyeleri firma görüşmelerinin firma yerinde yapılması ve sık bir şekilde yapılması yönündedir. Ayrıca firmalar sektörel farklılıkların bu teşvik ve desteklerde göz önüne alınmasını da talep etmektedirler. 5.2.5. Teşvik Bu tema finansman için teşvik, enerji için teşvik, vergi teşviği ve yatırım teşviği kategorilerinin bir araya gelmesi ile meydana gelmiştir. Ayrıca alt kategoriler arasında tek tip KDV uygulaması ve teşvik bilgilendirme faaliyetleri de bulunmaktadır. Araştırmaya katılan komite üyeleri sektördeki firmaların yaşadığı finansman sorunlarına verdiği cevaplarda KOBİ’ler için üretim alanında uzun vadeli kredi teminin kolaylaşması gerektiği cevabını vermişlerdir. Araştırmaya katılan komite üyeleri sektördeki firmaların yaşadığı finansman sorunlarına verdiği cevaplarda KOBİ’ler için finansman ihtiyacının yanında üretim alanında hizmet veren firmalara yüksek maliyete sebep olan enerji kullanımı alanında teşvik sağlanması gerektiği cevabını vermişlerdir. Araştırmaya katılan komite üyeleri sektördeki firmaların yaşadığı finansman sorunlarına verdiği cevaplarda KOBİ’ler için üretim alanında hizmet veren firmalara hammadde temininde tek tip KDV uygulamasının olması ayrıca sektörel farklılıklara göre de vergi teşviği uygulamasının olması yönünde cevap vermişlerdir. Araştırmaya katılan komite üyelerinin sektördeki firmaların yatırım ve ihracat teşviklerinden faydalanma durumunun olup olmadığı ile ilgili soruya verdikleri cevaplar sektörel farklılar sebebiyle farklı yönde olmuştur. Kimi sektörde yatırım ve ihracat teşvikleri hakkında yeterli bilgi olmadığını kimi sektörlerde ise sıkı takip edildiği ve kullanıldığını da belirtmişlerdir. Ayrıca bu teşviklerden ticaret odasının firmaları bilgilendirdiğini de belirtmişlerdir. 296 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme 5.2.6. Beklentiler Bu tema Üniversite, OSB ve Ticaret Odası ile ilgili beklentilerinin sorulmasıyla eğitim, iletişim ve işbirliği kategorilerinin bir araya gelmesi ile meydana gelmiştir. Özellikle bu kurumlarla Üniversite- Sanayi işbirliği içerisinde iletişimin çok iyi olduğunu belirtmişlerdir. Bir Katılımcı: “Düzce Ticaret Odası ve Üniversite işbirliğinden memnunum. Bizde bireysel katkı vermeye çalışıyoruz. Bilimle paranın bir araya gelmesi çok önemli, bir tarafta bilgi var harekete geçmiyor bir tarafta para var, birleşme büyüme çok önemli, büyük oranda başarıldı. Rektör hanımda her konuda çok destek veriyor. Teori ve pratiğin karşılıklı bir araya gelmesi çok kıymetli, hayata geçirilmesi, çok önemli çok kıymetli, yapılan çok şey var daha da iyi olacak” Bu kurumlardan beklenti işletmelerin ihtiyaçlarına göre kalifiye elamanın yetiştirilmesi, işletmelere verilen eğitimlerin cezbedici hale getirilerek katılımların artırılmasının sağlanmasını talep edilmektedir. Ayrıca eğitimlerin bir pazar araştırması sonucu olarak yapılması, sektörel farklılıklar ve sektörel ihtiyaçlar göz önüne alınarak eğitimlerin düzenlenmesi gerekliliği üzerinde durulmuştur. “Eğitim alanlarında iki günlük sertifika değil de ciddi işin sonunda sınavlı eğitimler hazırlanmalı. Eğitimcinin de bir pazarlamacı gibi dükkân dükkân gezerek potansiyel öğrencisini bilerek araştırmasını yaparak, esnafı tanıyarak eğitimi planlaması, hastanın ne olduğunu tanımadan tedavi edemezsiniz, eğitiminde planlanması gerekmektedir.” Sonuç KOBİ’ler ülke ekonomilerinin dinamizmini sağlayan küçük yapı taşlarıdır. Bu işletmeler kuruldukları andan itibaren en başta istihdam yaratma olanağı sağlamaları, küçük bir eğitim kurumu gibi bünyelerinde kalifiye eleman yetiştirebilmeleri, ulusal/uluslararası şirketlere ara mamul üretmeleri, daha çok emek yoğun faaliyet yürütmelerinden kaynaklı değişimlere açık olmaları ekonomi için oldukça önem arz etmektedir. Yalnız bu işletmelerin kurulduktan sonra ihtiyaç duydukları rekabet edebilme gücü ve finansman ihtiyaçlarını karşılayabilme çabaları önlerinde oldukça zorlu engeller çıkarabilmektedir. Düzce ilinden yer alan KOBİ’lerin yaşadığı pazarlama ve finansman sorunlarını öğrenebilmek için görüşmeler meslek komite toplantıları ya da meslek komite temsilcileri ile gerçekleşmiştir. Sektör temsilcileri ile yapılan görüşmeler yarı yapılandırılmış görüşme tekniği ile hazırlanan sorularla yüzyüze görüşerek 1-3 saat aralığında sürelerle gerçekleşmiştir. Görüşmeler 297 Serhat Ata / Ali Özer / Nevin Özer \| kendilerinden izin alınarak ses kaydına alınmış ve yazıya çevrilmiştir. Elde edilen cevaplar doğrultusunda temalar ve alt kategoriler oluşturulmuştur. Sektör temsilcileri ile pazarlama sorunları açısından yapılan görüşmelerde müşteri ilişkileri, risk yönetimi, satış ve pazarlama yönünden eksikliklerinin olduğu, bu konuda kendilerinin işgören yetiştirseler bile deneyim kazanan işgörenin büyük firmalara yönelmesi ve dijital dünyaya ayak uydurma konusunda eğitime ihtiyaç duydukları yönünde oluşmuştur. Ayrıca eğitim konusunda iki farklı görüş ortaya çıkmıştır. Birincisi ihtiyaç duyulan bu konularda Düzce Ticaret Odası ve KOSGEB tarafından eğitimlerin verildiği ama günlük iş akışı sürecince yetişemediklerini ifade etmişlerdir. İkincisi ise bu konularda verilen eğitimlere ihtiyaç duymama ve bu yüzden de eğitimlere katılmama söz konusu olmaktadır. Bunun sebebinin olarak sektörel farklılıklar sebebiyle ortaya çıktığı düşünülmektedir. Satış personeli becerileriler ile ilgili tecrübe eğitim ve pazarlayabilme bilgisi eksikliği yine firmaların birçok pazarlama sorununun en önemli gerekçelerindendir. Söz konusu problem personelin müşteri ile ilişkilerini etkileyebildiği gibi temel pazarlama karmasında yaşanan sorunlarda da etkili olabilmektedir. Bu noktada yapılması gereken en önemli şey her ne kadar firmalara külfet getirdiği düşünülse de personeli pazarlama eğitimi, oryantasyon ve sertifika sağlayan programlara göndermek veya işverenin ön koşul olarak bu eğitimi talep etmesi bir nebze de olsa sorunun çözümüne yardımcı olacağı düşünülebilir. KOBİ’lerin pazarlama sorunlarının enflasyon ve yarattığı olumsuz şartlar etrafında şekillendiği söylenebilir. KOBİ’ler ekonominin içerisinde en küçük birim olmakla birlikte enflasyonun yarattığı her olumsuzluktan da anında etkilenen birim konumundadır. KOBİ’lerin rekabet gücü, promosyon uygulamada daha esnek olabilmeleri, büyük firmaların daha teknoloji yoğun olmaları ve dijitalleşmeye hızlı ayak uydurabilmeleri, nitelikli işgörenlerin büyük firmalara yönelmesi ve ürün farklılaşmasına büyük firmaların daha rahat gidebilmeleri KOBİ’lerin önünde oldukça olumsuz hava yaratmalarına neden olmaktadır. Sonuç Ayrıca hammadde fiyatları, ikame ürün sıkıntısı, fiyat kaynaklı pazarlamanın tüm karması, stok maliyet artışı, reklam bütçelerinin düşmesi, fuar maliyetlerinin artması, akaryakıta dayalı lojistik maliyetlerin artması büyük sorundur. Söz konusu sorunların giderilmesinde mevcut ekonomik koşullarda devletin desteği geçici çözüm sağlamada etkili olabilir. Her ne kadar coğrafi açıdan Düzce’deki KOBİ’ler oldukça iyi bir konumda olsalar da rekabetin yoğun yaşanabilme ihtimali bir nebze de olsa KOBİ’leri etkilemektedir. Rakipler ve büyük firmalar sektöre giriş yaptığında çok kolay adapte olup KOBİ’lerin sahip olduğu müşterileri hedef alabilmektedir. Söz konusu rakiplerin pazara bu kadar kolay hâkim olabilmeleri coğrafi konum 298 Düzce’deki Kobi’lerin Pazarlama ve Finansal Sorunları Üzerine Bir Değerlendirme ile alakalı olduğu kadar KOBİ’lerin ürünün zamanında teslim edilememesi, ikame ürün bulunduramama, kaliteye dayalı şikâyetler, kötü müşteri ilişkileri yönetimi, garanti kapsamında müşteri şikâyetleri gibi temel pazarlama karması sorunlarından kaynaklanabilmektedir. Yapılan görüşmeler sonucunda teknoloji açısından KOBİ’lerin yeni dijital çağa kendilerini adapte edemeyip yenilikleri takip edemedikleri görülmektedir. Bu sorunda bireysel dijital satıcıların artışı, kuşak farklılıklarına dayalı teknolojik sorunlar yenilikleri takip açısından KOBİ’leri etkilemektedir. Bu noktada mutlaka KOBİ’lerin dijital temsilci veya asistan gibi uygulamaları firmalara adapte etmeleri çözüm sağlamada etkili olabilecektir. Finansman açısından sorulan sorulara verilen cevaplarla oluşturulan temalar ve alt temalar; Sektör temsilcilerinin finansman sorunları sorusuna verdikleri cevaplarla oluşturulan alt başlıklar özkaynak yetersizliği, maliyet yüksekliği, taksit ödeme sıkıntısı, istenilen kredi miktarına ulaşılamama, kredi verenlerin isteksizliği, finansmana ulaşmada zorluklar olmaktadır. Sektör temsilcilerinin yaşanılan finansman sorunları için önerileri sorusuna verdikleri cevaplarla oluşturulan alt başlıklar vergisel teşvik, düşük faizli kredi, sektörel teşviklerin artması, firma kotası, kar skalalarının artırılması, iflas riski, tek tip KDV, kredi takibi, sektörel kredi kolaylığı olmaktadır. Sektör temsilcilerinin muhasebe ve finansman departmanlarının varlığı sorusuna verdikleri cevaplarla oluşturulan alt başlıklar departmanlaşmanın olmaması, muhasebe ve finans konularında karar vericinin yönetici olması olmaktadır. Sektör temsilcilerinin kurumların sunduğu teşvik ve danışmanlık hizmetinden yararlanabilme sorusuna verdikleri cevaplarda sektörel olarak farklılık göstermekte, kimi sektör temsilcileri yeterli görürken kimi temsilerin kendi sektörleri için yeterli görmemektedir. Sektör temsilcilerinin finans kuruluşlarından yararlanma sorusuna verdikleri cevaplarla oluşturulan alt başlıklar banka kredisi ve çeşitleri, yatırım bankası kredisi, KGF desteği olmaktadır. Sektör temsilcilerinin finansal teknikler bilgi ve kullanma durumu sorusuna verdikleri cevaplarla oluşturulan alt başlıklar Leasing ve Barter oluşturmaktadır. Sektör temsilcilerinin yatırım ve ihracat teşviklerinden yararlanma sorusuna verdikleri cevaplarla oluşturulan alt başlıklar yatırım teşvikleri ve ihracat teşvikleridir. Sektör temsilcilerinin OSB, Ticaret Odası ve Üniversiteden beklentileri sorusuna verdikleri cevaplarla oluşturulan alt başlıklar kalifiye eleman yetiştirilmesi, kurumlarla iyi iletişim, piyasa araştırmasıyla belirlenen eğitim isteği, sertifikalı eğitim olarak belirlenmiştir. Sonuç Finansman ile ilgili temalar değerlendirildiğinde Düzce ilinde yer alan KOBİ’lerin organizasyonel yapısında muhasebe ve finans departmanlarının yer almadığı, bu konularda uzman kişi yerine şirket yöneticisinin karar aldığı 299 Serhat Ata / Ali Özer / Nevin Özer \| görülmektedir. KOBİ’lerin genel olarak ticari bankalarla aktif ilişki içerisinde oldukları ve finansman arayışlarında ilk sırada yer aldıkları tespit edilmiştir. Bir sektörde leasing işlemi ile finansman sağladıkları yine iki sektörde ticari işlemleri yürütürken ikili ilişkilere dayalı Barter işleminin kullanıldığı anlaşılmıştır. KOBİ’lerin finansman sıkıntılarının literatürde yer alan genel KOBİ finansman sıkıntıları ile aynı olduğu tespit edilmiştir. Düzce ilinde yer alan KOBİ’ler yatırım ve ihracat teşviklerinden yararlanmaktadırlar. Beklentileri ise sektörel finansman kolaylıkları, vergi kolaylıkları, kalifiye elemanın yetiştirilmesidir. 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Sosyal Ekonomik Araştırmalar Der- gisi, 1(1-2), 187-204. Bölüm 16 Kâr Payı Dağıtımı: Politika ve Teoriler Kâr Payı Dağıtımı: Politika ve Teoriler Kâr Payı Dağıtımı: Politika ve Teoriler Ömer Kaya2 Özet Finansal yönetimin temel karar alanları finansman, yatırım ve kâr payı dağıtımına yönelik konuları içermektedir. Kaynakça Günümüzde şirketlerin temel amacı olan hem risk hem zaman faktörünü dikkate alan firma değerinin maksimize edilmesi hissedarların servetlerinin maksimize edilmesini sağlayacak şekilde kararlar alınması ile mümkün olmaktadır. Şirketlerin hem büyümeye engel olmayacak aynı zamanda hissedarların beklentilerini karşılayacak kâr payı dağıtımı politikası benimsemeleri son derece önemlidir. Kâr payı dağıtımının firma değeri üzerindeki etkisi konusunda farklı teoriler bulunmakla beraber konu ile ilgili iki temel teori Modigliani ve Miller tarafından ileri sürülen kâr payı dağıtımının firma değerini etkilemediği görüşü ile Lintner ve Gordon’un savunduğu kâr payı dağıtımının firma değeri üzerinde etkisi olduğu görüşüdür. Ancak piyasanın ve ekonominin sürekli değişen dinamikleri nedeniyle her şirket için ideal bir kâr payı dağıtım politikasından bahsetmek mümkün değildir. 1 Dr. Öğr. Üyesi, Bingöl Üniversitesi, ekaya@bingol.edu.tr, ORCID ID: 0000-0002- 1823-162X 2 Öğr. Gör., Sakarya Üniversitesi, omerkaya@sakarya.edu.tr, ORCID ID:0000-0002- 1211-0341 303 1 Dr. Öğr. Üyesi, Bingöl Üniversitesi, ekaya@bingol.edu.tr, ORCID ID: 0000-0002- 1823-162X 2 Öğr. Gör., Sakarya Üniversitesi, omerkaya@sakarya.edu.tr, ORCID ID:0000-0002- 1211-0341 https://doi.org/10.58830/ozgur.pub401.c1764 1. Giriş Günümüz şirketlerinin temel amacı firma değerinin maksimizasyonunu sağlamaktır. Söz konusu amaç şirketin hisse senetlerinin piyasa değerinin, diğer bir ifadeyle hissedarların servetlerinin, maksimize edilmesini sağlayacak şekilde kararlar alınması ile mümkün olmaktadır. Değer maksimizasyonunun sağlanmasına etki eden en önemli faktörlerden biri kâr payı dağıtım politikasıdır. Kâr bir şirketin toplam gelir ve giderleri arasındaki pozitif farkı; kâr payı ise söz konusu pozitif farktan yasal karşılıkların çıkarılmasıyla birlikte hissedarlar arasında yapılan yatırım ile orantılı bir şekilde dağıtılmasını ifade etmektedir. Kâr payı dağıtım politikası dönem sonundaki net kârın ne https://doi.org/10.58830/ozgur.pub401.c1764 303 304 Kâr Payı Dağıtımı: Politika ve Teoriler kadarının şirket bünyesinde oto finansman olarak bırakılacağını, ne kadarının hissedarlara dağıtılacağına yönelik kararları içermektedir. Şirketler ileriye dönük hedef ve amaçları doğrultusunda detaylı ve doğru analizler yaparak bu kararlar arasında seçim yapmaktadırlar. Ancak piyasanın ve ekonominin sürekli değişen dinamikleri nedeniyle her şirket için ideal bir kâr payı dağıtım politikasından bahsetmek mümkün değildir. Bu araştırmada öncelikle teorik çerçevede kâr payı dağıtım politikaları ve bu politikaları etkileyen faktörler ele alınmış, devamında kâr payı dağıtım politikası türleri ve dağıtım teorileri açıklanmıştır. 2. Kâr Payı Dağıtım Politikaları Kâr payı dağıtım politikası, elde edilen kârın ne kadarının ortaklara ödeneceğini belirler. Kâr payı dağıtımı şüphesiz ki işletmenin dağıtılmamış kârlarını azaltacak ve oto finansman miktarını etkileyecektir. Kâr payı dağıtım politikaları ortakların servetini maksimize etme çabaları içinde de önemli bir yer tutmaktadır. Çünkü ortakların serveti, hisselerin piyasa değerinden ve dağıtılan kârlardan oluşmaktadır. Kâr payı dağıtım politikaları yatımcıların beklentilerini ve dolayısıyla davranışlarını etkileyerek, şirketin piyasa değerine bir etkide bulunur. Örneğin, ortaklar yüksek oranda kâr payı dağıtılmasını kârın firmada bırakılmasına kıyasla daha emniyetli buluyorlarsa kâr payı ödemesi yapılmasını istemeyebilir veya düşük oranlı kâr payı ödemesi yapılmasına olumsuz tepki gösterebilirler ve bu durum hisse senedinin piyasa fiyatının düşmesi sonucunu doğurabilir. Firmanın bunu engellemek için yüksek oranda kâr payı dağıtımına gitmesi, firmanın bünyesinde daha az kâr alıkonmasına ve şayet yatıranlar için ihtiyaç duyduğu fonları sermaye piyasasından tedarik etme imkânı yoksa büyüme hızının yavaşlamasına yol açar. Öte yandan tam tersi bir şekilde firma sermaye artırımı yoluyla kaynak sağlama olanağına sahip olsa bile ortaklar ödeyecekleri gelir vergisi ve sermaye artırmanın gerektireceği giderler nedeniyle kârın dağıtılmayarak firma bünyesinde bırakılmasını tercih edebilirler. Böyle bir durumda belki de yüksek oranlı kâr payı dağıtımı nedeniyle hisse senedi fiyatlarının düşmesi söz konusu olabilir. Bu söylenenler her firma için geçerli olabilecek ideal bir “kâr payı dağıtım politikası” nın söz konusu olamayacağını, kâr payı dağıtım politikalarının firmadan firmaya ya da koşuldan koşula değişebileceğini göstermektedir (Boztosun, 2006: 6-7). 3. Kâr Payı Dağıtım Politikasını Etkileyen Faktörler Teorik anlamda kâr payı dağıtım politikalarının amacı şirketlerin firma değerini en yükseğe çıkarmak olmakla birlikte işletmelerde kâr payı dağıtım 305 Emine Kaya / Ömer Kaya Emine Kaya / Ömer Kaya politikalarını etkileyen bir dizi faktörden söz etmek mümkündür. Bu faktörler şunlardır (Dendeş, 2020: 71-75): Ülkelerin geçerli yasaları veya şirketlerin ana sözleşmelerindeki hükümler, kâr payı dağıtımları üzerinde belirleyici olabilmektedir. Örneğin, ülkemizde anonim ortaklara kâr payı dağıtılmasına yönelik TTK ve SPK’nın bir takım düzenlemeleri bulunmaktadır. Ayrıca şirketlerin ana sözleşmelerinde yer alan hükümlerle kârın belli bir bölümünün “statü yedekleri” şeklinde ayrılması veya belirli amaçlara tahsisi mümkün olmaktadır. Öte yandan şirketlere büyük miktarda borç veren ya da tahvillerini satın alan finansman kurumları, düzenlenen kredi sözleşmeleri ile kâr payı dağıtımını sınırlama olanağını elde edebilmektedirler. Şirketin likidite durumu, yüksek miktarda dönem kârı elde edilmesine ve/veya dağıtılmamış kârlar olmasına rağmen, kâr payı dağıtılamayabilir ve kâr payı nakit şeklinde dağıtılmayabilir. Ayrıca şirketlerin sermaye artırımına gitmeleri ve hissedarlara bedelsiz hisse sendi dağıtmaları yedek akçelerin sermayeye eklenmesi yoluyla mümkündür. Böylelikle nakit çıkışı olmaksızın kâr payı dağıtımı yapılabilmektedir. Büyüme sürecinde olan şirketler, büyümelerini gerçekleştirebilmek için daha fazla finansmana ihtiyaç duymaktadır. Şirket açısından borçlanmanın uygun bulunmadığı, yeniden hisse senedi halka arz etmenin ise zahmetli ve hissedarlar tarafından istenmemesi durumunda, şirket kârı dağıtılmayarak fon sağlanabilir. İşletmenin varlıklarının kullanımıyla sağlayacağı kârlılık oranı, kârın ortaklara dağıtılması halinde ortakların bu fonlan alternatif yatırım alanlarında değerlendirerek elde edebilecekleri kârlılık oranından yüksekse, kârın dağıtılmaması ortaklar açısından daha cazip olabilir. Özellikle kârın dağıtılmaması, hisse senetlerinin önemli oranda değer kazanmasına yol açacaksa, ortaklar bunu tercih edebileceklerdir. Bir firmanın borç yükünün hayli yükselmiş olduğu durumlarda, ödeme zamanı gelmiş borçların tekrar borçlanma şeklinde ödenmesi yerine, faaliyetleri sonucu yaratmış olduğu fonların kullanılmasıyla kapatması ve borcun azaltılması daha akılcı olabilecektir. Böyle bir durumda elde edilen kârın dağıtılması yerine borç ödemede kullanılması tercih edilebilir. İşletmelerin uzun vadeli fon gereksinmelerini en uygun koşullarla karşılayabilecekleri yer sermaye piyasalarıdır. Ancak sermaye piyasasına başvurma olanağı, şirketlerin ölçeğine ve piyasadaki imajına bağlı olarak değişebilmektedir. Yeterli büyüklüğe sahip veya düzenli bir büyüme süreci içine girmiş, istikrarlı, tanınmış firmalar, sermaye piyasasından daha Kâr Payı Dağıtımı: Politika ve Teoriler 306 kolaylıkla fon temin edebileceklerinden, kâr payı dağıtım oranlarını da daha yüksek belirleyebilirler. Buna karşılık, sermaye piyasalarına erişme imkânları kısıtlı olan firmalar, yatırımları için oto finansman kaynağına daha büyük ihtiyaç duymakta, dolayısıyla kâr payı dağıtım oranlarını daha düşük tutmak zorunda kalmaktadırlar. İşletmede dağıtılmayıp işletme bünyesinde tutulan kârların, dağıtılan kârlara oranla daha düşük oranda vergilendirilmesi düşük oranlı kâr payı dağıtımını teşvik edici bir faktör oluşturabilmektedir. 4.1. Miktar Olarak Kâr Payı Dağıtım Politikaları İşletmelerin her yıl sabit bir miktarda ya da sermayelerinin sabit bir oranında kâr elde etmeleri mümkün olmamakla birlikte işletmeler kâr payı dağıtımlarında belli bir düzen içinde davranma ve bu şekilde ortaklarının da güvenini kazanma ihtiyacı duyabilirler. İşletmelerin dağıtacakları kâr payının miktarını belirlemek için kullandıkları birkaç yöntem bulunmaktadır. 3. Kâr Payı Dağıtım Politikasını Etkileyen Faktörler Öte yandan, ortakların yüksek gelir diliminde bulunması ve elde edecekleri kâr payı gelirleri üzerinden ödeyecekleri vergi oranının, kârın işletme bünyesinde tutulması sonucunda oluşabilecek sermaye kazançları üzerinden ödenecek vergi oranlarından yüksek olması da düşük kâr payı dağıtımının tercih edilmesini sağlayabilmektedir. Genel itibariyle aile şirketleri veya az ortaklı, hisseleri halka arz edilmemiş şirketler, şirketleri ya da belli bir gruba ait şirketlerde, yönetimde söz sahibi olanların şirket üzerindeki kontrolü elden kaybetmekten korkmaktadır. Bu sebeple yatırımlar için ihtiyaç duyulan fonların tedariğnde ortak sayısının artmasına yol açabilecek hisse senedi halka arz etmek gibi yöntemler yerine, oto finansman fonlarına ağırlık vermek, dolayısıyla da kâr payı dağıtımını düşük tutmak eğilimi ağırlık kazanmaktadır. 4. Kâr Payı Dağıtım Politikası Türleri 4.1.1. Sabit Miktarda Kâr Payı Dağıtım Politikası Bu politika kâr payı dağıtımında yıllar itibarı ile aynı miktarı muhafaza etmeyi hedeflemektedir. Elde edilen kâr tutarı 1 yıl öncesine kıyasla düşük ya da yüksek olsa dahi dağıtılan kâr payı tutarı sabit kalmaktadır. Sabit miktarda bir kâr tutarının dağıtımı, ileriye yönelik belirsizliği azaltması nedeniyle ortaklar açısından çekici bulunmaktadır. Özellikle yatırımlarından sağladıkları geliri önemseyen yatırımcılar, kâr payı dağıtımını sabit bir miktar olarak yapan şirketleri tercih etmektedir. Ancak böyle bir politika, hisse senetlerini sabit getirili bir yatırım aracına dönüştürmektedir. 307 Emine Kaya / Ömer Kaya Emine Kaya / Ömer Kaya \| Her yıl sabit tutarda kâr payı dağıtımı, hisse senetlerinin piyasa fiyatlarının artması konusunda sınırlayıcı bir etkiye sahip olmakta, bu şekilde bu tür kâr payı dağıtımı yapan şirketlerin hisse senetlerinin fiyatı yükselmemektedir. Dolayısıyla böyle bir politika, sermaye kazancı sağlamak isteyen yatırımcılar açısından cazip olmamaktadır (Kaya, 2009: 43). 4.1.3. Sabit Miktar veya Oran + İlave Kâr Payı Dağıtım Politikası Bu politikada, şirket asgari bir kâr dağıtım miktarı ya da oranı saptamakta, elde edilen kârın yüksek olduğu dönemlerde bu miktar veya oranın üzerine ilavede bulunulmakta ve her yıl dağıtılan kâr payı, belirli bir miktar ya da oranın altına düşmemek kaydıyla dalgalı bir seyir göstermektedir. Böylece ortaklar için belirli bir gelir garantilenirken, şirketin kârına bağlı olarak bazı yıllarda daha yüksek kâr payı elde edilebilmektedir. 4.1.2. Sabit Oranlı Kâr Payı Dağıtım Politikası Bu politikaya göre işletmeler, şirket kârının belli bir yüzdesini dağıtır. Ortaklara dağıtılan kâr payları, elde edilen kâr değiştikçe değiştiğinden, bu politikayı benimseyen işletmeler bazen çok düşük bazen de çok yüksek kâr payı dağıtabilmektedirler. Şirket yönetimini büyük hissedarlara bırakan küçük hissedarların, şirket kârından faydalanmalarını garantiye almak amacıyla şirket sözleşmelerinde sabit oranlı kâr payı dağıtımı olacağına dair maddeler bulunabilmektedir. Ancak kâr dağıtımında bu şekilde bir politikanın benimsenmesi, kâr rakamlarının azaldığı ya da faaliyetler sonucunda zarar elde edildiği durumlarda sorunlar yaşanabilmektedir (Özaltın, Ersoy ve Bekçi, 2015: 400). 4.2. Ödeme Şekli Açısından Kâr Payı Dağıtım Politikaları Ödenecek miktar açısından olduğu gibi, ödemenin şekli açısından da kâr payı dağıtımları farklılaşabilmektedir. 4.1.4. İstikrarlı Kâr Payı Dağıtım Politikası Şirketler genellikle kâr payı dağıtım oranlarını, ortaklarına istikrarlı bir gelir sağlayacak şekilde düzenlemeye çalışmaktadırlar. İstikrarlı kâr payından kasıt, kâr paylarının zaman içinde mümkün mertebe azalma göstermeksizin belli bir düzeyi muhafaza etmesi ve çok hızlı olmasa da yine belli bir artış trendini izlemeleridir. Genellikle böyle bir istikrar politikası izleyen firmaların hisse senedi fiyatlarının, sabit oranlı kâr payı dağıtan ancak kârlarının düzensizliği nedeniyle, kâr payı dağıtımları da istikrarsız seyreden firmaların hisse senetlerininkinden yüksek olduğu görülebilmektedir. Kararlı ve düzenli artan şekilde kâr payı dağıtımı gerçekleştirebilmek için firmalar aşağıdaki önlemleri alabilir: 308 Kâr Payı Dağıtımı: Politika ve Teoriler • Artan kârlarla beraber, kâr payı yüzdesini kademeli olarak arttırmak, artan kârın bütününü hemen dağıtmamak, • Kârlardaki yükseliş istikrarlı hale gelmeden, mevcut kâr payı dağıtım yüzdesini arttırmamak, • Kârlarda uzun bir dönem süren kayda değer azalışlar olmadan, kâr payı yüzdesinin değiştirmemek. 4.2.1. Kâr Payının Nakit Olarak Dağıtılması En çok rastlanan kâr payı dağıtım türü, ortaklara kâr payı bedellerinin para şeklinde ödenmesidir. İşletmenin likiditesi elverişli ise kâr payı, şirketteki hali hazırdaki mevcut nakitlerle ödenir. Hisse senetlerinin fiyatı daha önceden, dağıtılacak kâr payı miktarı kadar yükselmiş olacağından, kâr payı dağıtımına paralel hisse senetlerinin fiyatı da düşer. Dağıtımın yapılmasıyla beraber hissedarların mevcut durumunda herhangi bir değişiklik olmamakta, hissedarlar belli miktarda nakit elde ederken, aynı miktarda hisse senetlerinin değeri düşmektedir (Demirel, 2014: 37). Ancak işletmenin dönem kârının yüksek olması her zaman nakit ve nakit benzeri varlıkların da yüksek olacağı manasına gelmemektedir. Hızlı büyüyen işletmeler, elde ettikleri nakdin büyük bir kısmını varlıkların finansmanı için kullanabilirler. Ayrıca, herhangi bir büyüme veya genişleme faaliyeti olmadığı durumlarda bile yüksek enflasyonun işletme sermayesi üzerinde yıpratıcı etkisi görülmektedir. Böyle durumlarda rakamsal olarak işletme faaliyetlerinden elde edilen kâr yüksek olmasına rağmen kâr payını ödeyecek yeterli nakit bulunmayabilir. İşletmeler likidite kısıtının bulunduğu/ sermaye yapısının değiştirilmek istendiği ya da yatırımcıların vergi tasarrufu sağlayabilecekleri durumlarda, nakit kâr payı ödemek yerine alternatif yollara başvurabilirler. 4.2.2. Kâr Payının Bedelsiz Hisse Senedi Şeklinde Dağıtılması Hisse senedi şeklinde kâr payı dağıtımı, işletmenin daha önceleri firma bünyesinde alıkoymuş olduğu kârlarının bir kısmını sermaye hesabına aktararak sermaye artırımı gerçekleştirmesi ve arttırılan sermayeyi temsil eden yeni hisse senetlerim mevcut ortaklara hisseleri oranında, bedel karşılığı olmaksızın dağıtmasıdır. Emine Kaya / Ömer Kaya 309 Olağanüstü Yedekler= 80.000.000 TL Olağanüstü Yedekler= 80.000.000 TL Şirketin bedelsiz hisse senedi dağıtımı (%50) ile olağanüstü yedeklerinde bulunan 50 milyon TL’yi ödenmiş sermayesine eklediğini varsayarsak bilançonun görünümü aşağıdaki şekle dönüşecektir: Ödenmiş Sermaye(150.000 hisse1.000 TL)=150.000.000 TL Ödenmiş Sermaye(150.000 hisse1.000 TL)=150.000.000 TL Yasal Yedekler= 40.000.000 TL Yasal Yedekler= 40.000.000 TL Olağanüstü Yedekler=30.000.000 TL Olağanüstü Yedekler=30.000.000 TL Örnek: Bir işletmenin kâr dağıtımından önce bilançosunun öz sermaye bileşiminin aşağıdaki gibi olduğunu düşünelim: Ödenmiş Sermaye (100.000 hisse1.000 TL) = 100.000 000 TL Ödenmiş Sermaye (100.000 hisse1.000 TL) = 100.000 000 TL Yasal Yedekler= 40.000.000 TL Yasal Yedekler= 40.000.000 TL 4.2.3. Hisse Senetlerini Geri Satın Alma Yoluyla Kâr Payı Dağıtımı İşletmelerin bazen aşırı derecede nakde sahip olmalarına karşın, söz konusu nakdin yönlendirilebileceği kârlı yatırım olanaklarının bulunmaması, eldeki nakit mevcudunun piyasadaki hisse senetlerinin bir kısmının geri alınması amacıyla kullanılmasını cazip hale getirebilir. Bir işletmenin hisse sentlerini geri satın alması, kâr payı dağıtımının bir alternatifidir. Senetlerin satın alınması dolaşımdaki hisse senedi adedini azaltmakta, geri kalan hisseler için hisse başına kâr artış göstermektedir. Hisse başına karın artması, hisse senetlerinin piyasa değerini arttırmakta, ortaklar nakit kâr payı yerine sermaye kazancı elde etmiş olmaktadırlar. Ülkemizde bir anonim şirketin kendisine ait olan hisse senetlerini satın almasıyla dolaylı yolla kâr payı dağıtması hukuki açıdan mümkün değildir. Olağanüstü Yedekler=30.000.000 TL Bu işlem sonucunda hissedarların mevcut durumunda herhangi bir değişiklik oluşmamakta, sahip oldukları hisse senedi sayısı yükselirken, hisse senetlerinin fiyatı da eşit oranda azalmaktadır. Dikkat edilecek olursa bu işlem, kâr payının önce nakit olarak dağıtılması ve ardından işletmenin bedelli sermaye artırımına giderek ortaklarına nakit karşılığı hisse senedi satmasıyla aynı sonucu doğurmaktadır. Kâr payının hisse senedi şeklinde yatırımcılara dağıtılmasının bir takım yararları mevcuttur. Söz konusu yararlar (Akgüç, 1998: 792-793): • Şirketin hisse senedi şeklinde kâr payı dağıtımını nakit kâr payı dağıtımına tercih etmesi durumunda, ortakların elindeki hisse senedi sayısı artacaktır. Nakit paraya ihtiyacı bulunan ortaklar ellerine geçen hisse senedi fazlasını satarak paraya çevirebilir, bu şekilde mevcut hisselerini korurken aynı zamanda nakit geliri de yaratmış olurlar. Bu arada nakit paraya ihtiyacı olmayan ortaklar da hisse senedi yatırımlarını arttırmış ve daha fazla sayıda hisse senedi sahibi olacaklardır. • Kâr paylarının hissedarlara hisse senedi dağıtımı şeklinde olması vergi avantajı sağlamaktadır. Nakit kâr paylan gelir vergisine tabi tutulurken, hisse senedi şeklindeki kar paylarında, hisseler satılıncaya kadar verginin ertelenmesi ve sermaye kazancı olarak daha düşük vergilendirilmesi mümkündür. • Hisse senedi şeklindeki kâr payı dağıtımı genellikle büyüme potansiyeli içinde olan şirketlerde uygulanır. Bu nedenle bu yolla kâr payı dağıtımı yapan şirketler, yatırımcı açısından şirketin gelecek dönemlerdeki kazanç seviyesinde artış olacağı mesajını içermektedir. 310 Kâr Payı Dağıtımı: Politika ve Teoriler Ülkemizde bedelsiz hisse senedi dağıtımı sadece dağıtılmayan kârların değil aynı zamanda emisyon primi ve yeniden değerleme fonlarının sermayeye eklenmesi şeklinde de yapılmaktadır. Her ne kadar sermayeye ilave edilen fonların dağıtılmamış kârlardan gelmemesi durumunda, dağıtılan hisse senetlerini kâr payı olarak kabul etmek doğru değilse de, pratik olarak, yapılan bu işlemlerin de hisse senedi şeklindeki kâr payı dağıtımından önemli bir farkları olmamaktadır (Bolak, 2005: 271). Yabancı ülkelerde uygulanmakla birlikte, ülkemizde rastlanmayan bir diğer uygulama da, nominal değerlerinin azaltılması yoluyla hisse senetlerinin sayısının arttırılmasıdır. Burada şirket bilançosundaki öz sermaye kalemleri arasında herhangi bir aktarma ya da ödenmiş sermaye tutarında bir artış söz konusu değildir. Yapılan işlem sadece nominal değeri 1.000 TL olan bir hisse senedinin, örneğin, nominal değeri 500 TL olan iki adet hisse senediyle değiştirilmesidir. Finans literatüründe hisse senedi bölünmesinin kâr payı ödemesi olarak kabul edildiğini söylemek oldukça güç olmakla birlikte, bazı yazarlar hisse senedi bölünmesiyle, kâr payı dağıtımında bedelsiz hisse senedinden yararlanmanın hemen hemen aynı sonuçlara yol açtığını, dolayısıyla hisse senedi bölünmesinin de bir çeşit kâr payı dağıtımı olarak kabul edilebileceğini belirtmektedirler. 5.2. Kâr Payı Politikasının Geçerliliği Teorisi Gordon ve Lintner kâr payı dağıtım politikasının hisse senedi fiyatlarını dolayısıyla işletmenin piyasa değerini etkilediğini ileri sürmektedir. Eldeki kuş teorisi olarak da bilinen bu görüşe göre, nakit olarak dağıtılan kâr payları yatırımcıların gözünde, gelecekte gerçekleşecek ve belirsizlik içeren sermaye kazancından daha güvenli ve daha değerlidir. Diğer bir deyişle yatırımcılar bugün elde edilebilecek kâr payını ilerde elde edilebilecek bir sermaye kazancına tercih etmektedir (Aydın ve diğerleri, 2007: 426). 5. Kâr Payı Dağıtım Teorileri Hisse senetlerinin piyasa fiyatları üzerinde kâr payı dağıtım politikalarının etkisinin olup olmadığı varsa etkisi nasıl etkilediği konusu 1960’lardan bu yana finans alan yazınının önemli ilgi alanlarından birini teşkil etmiştir. Bu konu ile ilgili iki temel görüş Modigliani ve Miller’ın başını çektiği; kâr payı 311 Emine Kaya / Ömer Kaya Emine Kaya / Ömer Kaya dağıtım politikasının hisse senedinin değerini etkilemediğini öne süren görüş ile Lintner ve Gordon’un savunduğu kâr payı dağıtım politikasının hisse senedi fiyatı üzerinde etkili olduğu görüşleridir. Bu iki karşıt görüşün yanı sıra, kâr payı dağıtımının firma değeri üzerindeki etkisi konusunda değişik bakış açılan da bulunmaktadır. 5.1. Kâr Payı Politikasının Geçersizliği Teorisi Modigliani ve Miller’ın 1961 yılında yazmış oldukları makalelerinde savundukları görüşte hissedarlar elde edecekleri gelirin kâr payı veya sermaye kazancı şeklinde olması konusunda kayıtsızdırlar ve şirketin değerini yalnızca varlıklar ve yarattıkları gelirler belirlemektedir. Dolayısıyla ne kadar kâr dağıtıldığının şirket değeri üzerinde bir etkisi olmayacaktır. Uygun yatırım hacmini gerçekleştirmek isteyen işletme, yatırım için gerekli fonları ister elde ettiği kârlardan karşılasın, isterse elde ettiği kârın bir kısmını veya tamamım ortaklarına dağıtıp, gerekli fonları dağıttığı kâr payı kadar yeni hisse senedi ihracıyla karşılasın, iki seçenek de hisse senedi fiyatı üzerinde aynı etkiyi yaratacaktır. M-M tarafından geliştirilen bu bakış açısı, sermaye piyasasının mükemmel olduğu, işlem komisyonu, ihraç gideri ve vergi gibi giderlerin bulunmadığı, yatırımcıların daima akılcı davranışlar sergiledikleri ve gelecekte elde edilecek kâr paylarının tam belirlilikle bilindiği gibi varsayımlar içermektedir. Gerçek hayatta bu varsayımların gerçekleşmesinin son derece zor olduğu görülebilmekte, ayrıca verginin ihmal edilmesinden vazgeçilip, sermaye kazançtan üzerinden alınan verginin gelir vergisinden daha düşük olduğu kabul edildiğinde, kâr payı dağıtılmasının hisse senedi değerini azaltacağı sonucuna ulaşılmaktadır (Ertaş ve Karaca, 2010: 60). 5.4. Bilgi İçeriği ve Sinyalleme Hipotezi Şirketlerin gelecekte düzenli ve yüksek net kâr artışı veya kârlarında kesin bir düşüş tahmin etmedikçe kâr paylarını azaltmayacakları finans dünyasında genel kabul görmüş bir görüştür. M-M de bu görüşten yola çıkarak yatırımcıların, şirketlerin kâr paylarında yaptığı değişikliği yönetimin ilerdeki kârlarla ilgili olumlu ya da olumsuz sinyali olarak gördüklerini ileri sürmüşlerdir. Miller ve Rock’un ileri sürdüğü ve yukarıdaki görüşü destekleyen “asimetrik bilgi” kuramına göre yöneticiler şirketin kârlarının reel durumu ve ilerdeki yönüne dair, şirket dışındaki yatırımcılara kıyasla daha fazla bilgiye sahiptir. Bu sebeple kâr payı bildirimleri üzerinden şirketin durumu hakkında çıkarımda bulunmaları muhtemel ve anlaşılırdır. Buna göre (Bolak, 2010: 273-274). • Şirket ilk kez kâr dağıtır veya kâr payını yükseltirse, bu durum olumlu bir haber olarak algılanır ve şirketin piyasa değeri artar. • Şirket kâr payını düşürür ya da kâr dağıtımı gerçekleştirmezse, bu durumu olumsuz bir haber olarak yorumlanır ve şirketin piyasa değeri düşer. • M-M’e göre, yüksek kâr payı açıklayan şirket hisselerinin fiyatının artması, yatırımcıların yüksek kâr payı dağıtım oranını tercih etmeleri ile değil açıklanan yüksek kâr payının ileriye dönük olumlu sinyaller taşıması ile ilgili olmaktadır. 5.3. Artık Fonlar Teorisi Kâr payı dağıtım kararı, bu görüşe göre şirketlerin sermaye yapısı ve maliyeti belirlendikten sonra otomatik olarak oluşacak pasif bir karardır. İşletme r>k (getiri oranı sermaye maliyetinden büyükse), yani yatırımın net bugünkü değeri pozitifse sermaye yapısını koruyarak yatırım imkânlarını 312 Kâr Payı Dağıtımı: Politika ve Teoriler elindeki fonlarla karşılayacak, yatırım yapanların marjinal getirisi sermaye maliyetinden küçük olduğunda başka bir ifadeyle net bugünkü değeri negatif bir yatırım olduğunda elindeki fonları kâr payı olarak dağıtacaktır. Çünkü hisse sahipleri daha yüksek getiri elde edebileceklerine inandıklarında hakettikleri kâr payından vazgeçip şirket tarafından bu kâr paylarının değerlendirilmesini kabul etmektedir. Yatıranlardan beklenen getiri oranının sabit bir değeri varsa, en iyi çözüm, bu değer sermaye maliyetinin üzerinde olduğu müddetçe kâr payı dağıtım oranını “0”, altında olduğu sürece kâr payı dağıtım oranını “100” olarak belirlemektir. Beklenen getiri sermaye maliyetine eşit olduğunda dağıtılan kâr payı miktarının önemi kalmayacaktır (Brigham, 1986: 541). 5.5. Müşteri Grubu Etkisi Piyasada farklı kâr payı dağıtımı ihtiyacı hisseden yatırımcılar bulunabilir. Örneğin, yüksek vergi oranına tabi yatırımcılar düşük kâr payını tercih 313 Emine Kaya / Ömer Kaya Emine Kaya / Ömer Kaya ederken, düzenli nakit akışı isteyen küçük tasarruf sahipleri ya da emekli sandığı gibi kurumsal fonlar da yüksek kâr payını arzulayabilirler. Dolayısıyla değişik kâr payı dağıtım stratejilerinin değişik müşteri grupları bulunabilir ve şirketler hedefledikleri yatırımcı grubunu çekebilmek için kâr payı dağıtım politikalarını buna göre belirleyebilirler. Öte yandan herhangi bir şirket kâr payı dağıtım politikasında bir değişiklik yaptığında, borsada bu şirketin hisselerinin hareketlilik kazandığı gözlenebilecektir. Bunun nedeni, yeni stratejiden memnun olmayan hissedarların hisselerini elden çıkarmaya, bu stratejiye pozitif bakan diğer yatırımcıların ise hisse almaya çalışmalarıdır. 6. Sonuç Finansal yönetimin temel karar alanları finansman, yatırım ve kâr payı dağıtımına yönelik konuları içermektedir. Kâr payı dağıtım politikası, dönem sonundaki net kârın ne kadarının şirket bünyesinde oto finansman olarak bırakılacağını ne kadarının hissedarlara dağıtılacağına yönelik kararları içermektedir. Günümüzde şirketlerin temel amacı olan firma değerinin en yükseğe çıkarılması, hissedarların servetlerinin maksimize edilmesini sağlayacak şekilde kararlar alınması ile mümkün olmaktadır. Şirketlerin hem büyüme hem yüksek kâr payı dağıtımı amacı birbiriyle çelişmekte, dolayısıyla büyümeye engel olmayacak aynı zamanda hissedarların beklentilerini karşılayacak kâr payı dağıtımı politikası benimsemeleri son derece önemlidir. Kâr payı dağıtımının firma değeri üzerinde etkisi olup olmadığını, varsa etkinin nasıl olduğu konusu finans literatürünün önemli ilgi alanlarından birini oluşturmaktadır. Kâr payı dağıtımının firma değeri üzerindeki etkisi konusunda farklı teoriler bulunmakla beraber konu ile ilgili iki temel teori; Modigliani ve Miller’ın kâr payı dağıtım politikasının hisse senedinin değerini etkilemediğini öne süren görüşü ile Lintner ve Gordon’un savunduğu kâr payı dağıtım politikasının hisse senedi fiyatı üzerinde etkili olduğu görüşleridir. Ancak piyasanın ve ekonominin sürekli değişen dinamikleri nedeniyle her şirket için ideal bir kâr payı dağıtım politikasından bahsetmek mümkün değildir. 314 \| Kâr Payı Dağıtımı: Politika ve Teoriler 314 1 Dr. Öğretim Görevlisi, pasagultas@gmail.com, https://orcid.org/0000-0003-1215-3509 Özet Bu kitap bölümü, işletmelerin inovasyon ve sürdürülebilirlik arasındaki bağlantıyı anlamalarına ve gelecekte başarılı stratejiler geliştirmelerine yardımcı olmayı amaçlayarak kaleme alınmıştır. Paydaş katılımının ve beklentilerinin strateji belirlemedeki kritik rolü vurgulanarak, işletmelerin müşterilerden tedarikçilere kadar geniş bir paydaş kitlesinin beklentilerini anlamaları ve bu beklentilere uygun stratejiler geliştirmeleri önemli bir adım olarak öne çıkarılmıştır. Teknolojik inovasyonun sürdürülebilirlik stratejilerini şekillendirmedeki kilit rolü vurgulanarak, işletmelerin yenilenebilir enerji, enerji verimliliği ve çevre dostu üretim gibi inovatif çözümleri stratejik olarak entegre etmeleri gerektiği vurgulanmıştır. Sonuç olarak, bu bölüm, işletmelere gelecekteki sürdürülebilirlik hedeflerine ulaşmada yardımcı olacak önemli perspektifler ve uygulanabilir çözümler sunmaktadır. Kaynakça Akgüç, Ö. (1998). Finansal yönetim, İstanbul: Muhasebe Enstitüsü Yayınları. Aydın, N., Başar M., & Coşkun M. (2007). Finansal yönetim, Eskişehir: Türkiye Bilimler Akademisi. Bolak, M. (2010). İşletme finansı, İstanbul: Birsen Yayınevi. Bolak, M. (2005). İşletme finansı, İstanbul: Birsen Yayınevi. Boztosun, D. (2006). Temettü dağıtma kararının farklı piyasalarda hisse senedi fiyatına etkisinin mukayeseli olarak incelenmesi (Doktora Tezi). Gazi Üni- versitesi/Sosyal Bilimler Enstitüsü, Ankara). Brealey, R. A., Myers, S. C., Marcus, A. J., & Bozkurt, Ü. (1997). İşletme finans- manının temelleri, İstanbul: Literatür Yayıncılık. Brigham, E. F., (1986). Fundamentals of financial management, CBS College Publishing: New York. Demirel, E. (2014). Temettü dağıtım politikasının firma değeri üzerine etkisi ve BİST 30 Endeksinde bir uygulama (Yüksek Lisans Tezi). Selçuk Üniversi- tesi/Sosyal Bilimler Enstitüsü, Konya). Dendeş, E. (2020). Kâr payı dağıtım politikasını etkileyen mikro ve makro etkenler: Bist 100 firmaları üzerine bir uygulama (Yüksek Lisans Tezi). Aydın Ad- nan Menderes Üniversitesi/Sosyal Bilimler Enstitüsü, Aydın). Ertaş, F. C., & Karaca, S. (2010). Kâr dağıtımının ilanı ve gerçekleşmesi arasın- da geçen sürenin firma değerine etkisi. Muhasebe ve Finansman Dergisi, (47), 58-68. Kaya, T. (2009). Sermaye şirketlerinde kâr dağıtımının vergilendirilmesi ve muha- sebeleştirilmesi (Doktora Tezi). Dokuz Eylül Üniversitesi/Sosyal Bilimleri Enstitüsü, İzmir). Koç, P. (2021). Risk ve getiri arasındaki ilişkinin analizi: Borsa İstanbul’da işlem gören Türk futbol takımları üzerine bir araştırma. İşletme Araştırmaları Dergisi, 13(2), 1893-1906. Odabaşı, A. (2003). Sistematik risk tahmininde getiri aralığının etkisi: İMKB’de bir uygulama. Uludağ Üniversitesi İktisadi ve İdari Bilimler Fakültesi Der- gisi, 22(1), 107-120. Özaltın, S., Ersoy, E., & Bekci, İ. (2015). Kâr dağıtım politikasının bireysel ya- tırımcıların yatırım kararları üzerine etkisi. Süleyman Demirel Üniversitesi İktisadi ve İdari Bilimler Fakültesi Dergisi, 20(2), 397-411. Sevim, Ş., & Daşcı, H. (2020). Halka açık şirketlerde kâr dağıtım politikaları- nı etkileyen faktörler üzerine bir araştırma. İşletme Araştırmaları Dergi- si, 12(2), 1434-1446. Şeker, K., & Karabulut, R. (2018). Kâr payı dağıtım oranlarının borçlanma araç- ları üzerinde etkisi (Doktora Tezi). İktisadi Kalkınma ve Sosyal Araştırma- lar Derneği. Yalçıner, K. (2006). Risk ile getiri arasındaki doğrusallığın İMKB’de anali- zi. Muhasebe ve Finansman Dergisi, (29), 182-189. Bölüm 17 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri GİRİŞ İnovasyon ve sürdürülebilirlik, günümüz iş dünyasında öne çıkan ve işletmelerin stratejik yönetim süreçlerinde kilit bir rol oynayan iki temel kavramdır (Seebode ve Jeanrenaud, 2012). İnovasyon, organizasyonların rekabet avantajı sağlamak, yeni pazarlara ulaşmak ve sürdürülebilir büyüme elde etmek için stratejik olarak benimsediği bir süreçtir. Bu süreç, yeni ürün ve hizmetlerin geliştirilmesi, iş süreçlerinin iyileştirilmesi ve organizasyon kültüründeki değişimleri içermektedir (Maier vd., 2020). Diğer yandan sürdürülebilirlik, işletmelerin çevresel, sosyal ve ekonomik sorumluluklarına duyarlı bir şekilde hareket etmelerini ve uzun vadeli başarılarını sürdürmelerini hedefleyen bir stratejik yaklaşımdır (Adams vd., 2016). İşletmelerin inovasyon ve sürdürülebilirlik arasındaki derin etkileşimi anlamalarına ve bu iki kavramı entegre etmelerine yönelik bir çerçeve https://doi.org/10.58830/ozgur.pub401.c1765 315 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri 316 sunmak gerekmektedir. İnovasyonun sürdürülebilirlik üzerindeki etkisi, yeni teknolojilerin benimsenmesi, iş süreçlerindeki verimlilik artışları ve ürün yaşam döngüsü yönetimi gibi stratejik unsurları içermektedir (Avila vd, 2017). Aynı zamanda, sürdürülebilirliğin inovasyona sağladığı katkılar, çevresel etkilerin azaltılması, toplumsal değerlerin korunması ve etik standartlara uyum sağlama gibi unsurları içermektedir (Barbieri vd., 2010). Gelecekteki başarı için işletmelerin bu iki konsepti birleştirmeleri, küresel pazarlarda daha rekabetçi olabilmek, sürdürülebilirlik hedeflerine uyum sağlamak ve toplumsal beklentilere karşılık vermek açısından kritik bir öneme sahiptir (Gobble, 2012). Bu bağlamda, bu çalışmanın amacı, işletmelerin bu iki önemli paradigmayı birleştirerek sürdürülebilir başarı elde etmelerine nasıl katkı sağlayabileceğini detaylı bir şekilde incelemektir. 1. KAVRAMSAL BOYUT Günümüzde iş dünyası, sadece kârlılığı değil, aynı zamanda çevresel ve sosyal etkileri de göz önünde bulundurarak faaliyet gösterme zorunluluğuyla karşı karşıyadır. Bu bağlamda, sürdürülebilirlik ve yeşil inovasyon kavramları, şirketlerin geleneksel iş modellerini gözden geçirmelerini ve daha sürdürülebilir bir gelecek için çözümler üretmelerini sağlamaktadır. Sürdürülebilirlik, doğal kaynakların dengeli kullanımını, ekosistemlerin korunmasını ve toplumsal sorumlulukları içerirken; yeşil inovasyon, bu sürdürülebilirlik ilkelerini temel alarak yeni fikirler, ürünler ve iş süreçleri geliştirmeyi hedefler. Bu bölümde, sürdürülebilirlik ve yeşil inovasyonun temel prensiplerini anlamaya odaklanacak, çevresel, ekonomik ve sosyal boyutlarıyla açıklanmıştır. Yeşil inovasyonun ana ilkesi olan bu yaklaşım, iş dünyasının sadece bugünü değil, aynı zamanda yarını da düşünerek şekillenmesine rehberlik eden önemli bir stratejik yaklaşımdır. 1.1. Sürdürülebilirlik Sürdürülebilirlik, genellikle mevcut ihtiyaçları karşılamak için mevcut kaynakları kullanmanın yanı sıra gelecek nesillerin ihtiyaçlarını da göz önünde bulundurarak hareket etme ilkesini ifade eden bir kavramdır (Heinberg ve Lech, 2010; Higgins ve Larrinaga, 2014; Cambridge Dictionary, 2020). Doğal kaynakların sınırlı olduğu, iklim değişikliklerinin giderek arttığı bir dönemde, şirketlerin sürdürülebilirlik ilkelerine odaklanması büyük bir gereklilik haline gelmiştir. İş dünyasında sürdürülebilirlik, bir şirketin faaliyetlerini sürdürülebilir bir şekilde yönetmesi ve bu faaliyetlerin çevresel etkilerini minimize etmesi, toplumsal sorumluluklarına dikkat etmesi ve ekonomik olarak kârlı olması anlamına gelir (Boons vd., 2013). Paşa Gültaş \| 317 Paşa Gültaş \| 317 317 Paşa Gültaş \| 317 Sürdürülebilirlik, çevresel, ekonomik ve sosyal boyutları içeren geniş bir perspektifi içerir: Sürdürülebilirlik, çevresel, ekonomik ve sosyal boyutları içeren geniş bir perspektifi içerir: • Çevresel Sürdürülebilirlik: Çevresel sürdürülebilirlik, doğal kaynakların korunması, enerji verimliliği, atık yönetimi ve biyoçeşitliliği içeren çevresel etkilerin minimize edilmesi amacını taşır. İklim değişikliği, su kullanımı, hava ve su kirliliği gibi konuları içerir (Kuhlman ve Farrington, 2010). • Ekonomik Sürdürülebilirlik: Ekonomik sürdürülebilirlik, bir şirketin uzun vadeli kârlılığını ve sağlıklı büyümesini sürdürmesini ifade eder. Şirketin etik iş uygulamalarına uygunluk, finansal yönetim ve uzun vadeli stratejik planlamayı içerir (Ameer ve Othman, 2012). • Sosyal Sürdürülebilirlik: Bir şirketin toplumsal etkilerini yönetme kabiliyetini ifade eder. İş yerinde adil işe alım uygulamaları, çalışanların refahını artırmaya yönelik çabalar, topluluklara katkıda bulunma ve tüketici sağlığı gibi konular sosyal sürdürülebilirlik alanına dahildir (Husgafvel, 2015). Bu üç boyut arasındaki dengeyi sağlamak, sürdürülebilir bir iş modeli oluşturmanın anahtarıdır. Sürdürülebilirlik hem şirketlerin uzun vadeli başarılarını hem de küresel düzeyde çevresel ve toplumsal sorumluluklarını yerine getirmelerini hedeflemektedir (Epstein, 2018). Ayrıca sürdürülebilirlik yatırımlarının finansal performans üzerinde olumlu etkileri olduğu giderek daha fazla anlaşılmaktadır (Alshehhi vd., 2018). Yatırımcılar, çevresel ve sosyal faktörlere duyarlı şirketlere olan ilgilerini artırmaktadır. Bu da şirketlerin sürdürülebilirlik stratejilerini geliştirmelerini teşvik etmektedir (Ameer ve Othman, 2012). Sürdürülebilirliğin sadece bir eğilim değil, aynı zamanda iş stratejilerinin temel bir unsuru olmaya devam edeceği öngörülmektedir (Garetti ve Taisch, 2012). Şirketlerin çevresel ve sosyal sorumluluklarına duyarlı bir şekilde iş yapmaları, toplum ve dünya için olumlu bir etki yaratmaları, aynı zamanda uzun vadeli sürdürülebilir başarılarını güvence altına almalarına yardımcı olacaktır (Schaltegger, vd., 2012). Gelecekte sürdürülebilirlik, iş dünyasında daha belirgin bir rol oynamaya devam ederken, yeşil inovasyon bu evrilen paradigmanın özünde kilit bir rol üstlenmiştir (Singh vd., 2020). Yeşil inovasyon ve ar-ge çalışmaları, çevresel, ekonomik ve sosyal sürdürülebilirliği destekleyen çözümler sunarak, şirketlere rekabet avantajı sağlayacaktır (Chang, 2011). 1.1. Sürdürülebilirlik Ayrıca işletmelerin toplumsal sorumluluklarını daha etkili bir şekilde yerine getirmelerine olanak tanıyacaktır. 318 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri 1.2. Yeşil İnovasyon Yeşil inovasyon, sürdürülebilirlik çerçevesinde iş dünyasında ve toplumda çeşitli sektörlerde benimsenen bir yaklaşım olarak öne çıkmaktadır. Yeşil inovatif süreç, çevresel sorunların ele alınması, kaynakların sürdürülebilir kullanımı ve toplumsal sorumluluk ilkelerine odaklanarak işletmelerin ve endüstrilerin sürdürülebilir bir geleceğe adapte olmalarını sağlar (Chiou vd., 2011). Yeşil inovasyonun merkezinde, doğal kaynakların korunması ve atık üretiminin azaltılması gibi çevresel hedefler yer alır. Bu bağlamda, enerji verimliliği, yenilenebilir enerji kaynaklarına geçiş, malzeme kullanımının optimize edilmesi gibi konular, yeşil inovasyonun anahtar odak noktalarını oluşturur. İnovasyonun bu çevresel boyutu, şirketlerin ekolojik ayak izini minimize etmelerine ve çevresel sürdürülebilirlikle uyumlu ürün ve hizmetler geliştirmelerine olanak tanımaktadır (El-Kassar ve Singh, 2019). Yeşil inovasyonun ikinci önemli boyutu, ekonomik sürdürülebilirlik ilkesine dayanır. Bu, işletmelerin çevresel performanslarını artırırken aynı zamanda finansal açıdan etkin ve verimli olmalarını gerektirir (Singh vd., 2020). Yenilikçi iş modelleri, sürdürülebilirlikle uyumlu üretim süreçleri ve yeşil pazarlara odaklanma, şirketlerin rekabet avantajı elde etmelerini sağlar. Sosyal sorumluluk perspektifinden bakıldığında, yeşil inovasyonun üçüncü önemli bileşeni ortaya çıkar. İnovatif uygulamalar, işletmelerin toplumlarına ve paydaşlarına daha fazla katma değer sunmalarını hedefler. Adil iş uygulamaları, yerel topluluklarla iş birliği, eğitim ve fırsat eşitliği gibi sosyal konulara odaklanan inovasyonlar, şirketlerin toplumsal sorumluluklarını yerine getirmelerini sağlar (Wang vd., 2020). Yeşil inovasyon, tüm bu boyutları bir araya getirerek iş dünyasını daha sürdürülebilir, çevre dostu ve toplumsal olarak duyarlı bir geleceğe taşıma potansiyeline sahiptir. Bu bağlamda, şirketlerin yeşil inovasyonu benimsemeleri, sadece kendi sürdürülebilirlik hedeflerine ulaşmakla kalmaz, aynı zamanda küresel ölçekte çevresel ve toplumsal etkileri azaltma çabasına önemli bir katkıda bulunurlar (Bossle vd., 2016). Yeşil inovasyon, çevresel sürdürülebilirliği destekleyen ve iş süreçlerini daha verimli ve etkili hale getiren yenilikçi çözümleri içerir. Yenilikçi ürün tasarımları, çevre dostu üretim teknikleri ve enerji verimli iş uygulamaları, yeşil inovasyonun işletmelerin rekabet avantajını güçlendirmesine katkı sağlar. Sürdürülebilirlik eğilimleri aynı zamanda işletmelerin tedarik zincirleri üzerinde de etkili olmaktadır. Şirketler, tedarikçilerini daha sürdürülebilir Paşa Gültaş \| 319 Paşa Gültaş \| 319 319 Paşa Gültaş \| ve adil ticaret prensiplerine uygun hale getirme çabalarını artırmaktadır (Dangelico ve Pujari, 2010). Bunu başarmak, tedarik zincirinin her aşamasında çevresel ve sosyal sorumlulukların gözetilmesini sağlayarak, şirketlerin bu konuda daha şeffaf olmalarını gerektirir. Bu paradigmalar çerçevesinde, sürdürülebilirlik eğilimleri, şirketlerin sadece kâr amacı gütmekten öte, çevresel ve sosyal sorumluluklarına odaklanmalarını gerektiren bir dönüşümü yansıttığı söylenebilir (Cuerva vd., 2014). Bu eğilim hem iş dünyasını hem de toplumu olumlu yönde etkilemekte ve gelecekte daha sürdürülebilir bir iş modeli oluşturulmasına olanak tanımaktadır. 2. ÜRETİMDE SÜRDÜRÜLEBİLİRLİK Üretimde sürdürülebilirlik, işletmelerin çevresel etkileri azaltma ve kaynakları daha etkin kullanma amacını taşıyan bir dizi stratejiyi içermektedir. Yeşil üretim, bu stratejilerin temelini oluşturarak hem çevre dostu bir yaklaşımı benimser hem de işletmelere uzun vadeli rekabet avantajları sunar. 2.1. Yeşil Üretim Stratejileri Yeşil üretim stratejileri, küresel ölçekte işletmelerin sürdürülebilirlik hedeflerine ulaşmak için benimsediği kapsamlı yaklaşımları içerir (Song ve Yu, 2018). Bu stratejiler, çevre dostu uygulamaları, enerji verimliliğini artırmayı, atık azaltımını sağlamayı ve çevresel etkileri minimize etmeyi amaçlar. Uluslararası arenada bu stratejilerin uygulanması, küresel çapta çevresel sürdürülebilirliği destekleyerek ekonomik ve ekolojik dengenin korunmasına katkıda bulunur (Chen ve Chai, 2010). Enerji verimliliği, yeşil üretim stratejilerinin merkezinde yer alır. Uluslararası düzeyde enerji tüketiminin etkin bir şekilde yönetilmesi, iklim değişikliğiyle mücadelede önemli bir rol oynar. Şirketler, yenilenebilir enerji kaynaklarına geçiş, enerji tasarruflu teknolojilerin kullanımı ve üretim süreçlerinin optimize edilmesi gibi yöntemlerle enerji verimliliğini artırarak sürdürülebilir bir enerji altyapısı oluşturmayı hedefler (Cai vd., 2019). Atık azaltımı, küresel çapta doğal kaynakların korunması ve atık yönetiminin etkinleştirilmesi açısından kritik bir konudur. Uluslararası paylaşımlardan elde edilen en iyi uygulamalar, atıkların azaltılması, geri dönüşüm süreçlerinin iyileştirilmesi ve atık yönetimi stratejilerinin geliştirilmesi konusunda işletmelere rehberlik eder. Çeşitli sektörler arasında bilgi paylaşımı, etkili atık azaltımı ve döngüsel ekonomi konseptinin benimsenmesini destekler (Ilankoon vd., 2018). 320 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri Yeşil üretim stratejilerinin küresel çapta etkili olabilmesi için uluslararası iş birlikleri ve standartlar da büyük bir önem taşır. Uluslararası düzeyde sürdürülebilirlik standartlarının benimsenmesi, şeffaf raporlama ve paylaşılan çevresel performans ölçütleri, işletmelerin sürdürülebilirlik çabalarını uluslararası toplulukla uyumlu hale getirir (Ahmad ve Wu, 2022). Dolayısıyla yeşil üretim stratejileri, uluslararası paylaşımlardan elde edilen bilgi ve deneyimlerle desteklenerek küresel düzeyde sürdürülebilir bir üretim modelinin inşasına yönelik önemli adımları içerir (Li vd., 2022). Bu stratejiler, işletmelerin küresel sorumluluklarını yerine getirmelerini sağlamakla kalmaz, aynı zamanda küresel düzeyde çevre dostu bir endüstriyel dönüşümü teşvik eder. 2.2. Enerji Verimliliği Üretim süreçlerinde enerji verimliliği, küresel düzeyde sürdürülebilir üretim modellerinin benimsenmesi ve çevresel etkilerin azaltılmasına yönelik stratejik adımların atılmasını gerektiren önemli konulardır (Zakari vd., 2022). Enerji verimliliği, uluslararası düzeyde endüstriyel tesislerin enerji tüketimini optimize etmek amacıyla bir dizi yenilikçi çözümü benimsemesini sağlar. İşletmeler, enerji yoğun üretim süreçlerinde akıllı sensörlerin kullanımı, enerji tasarruflu teknolojilerin entegrasyonu ve yenilenebilir enerji kaynaklarına geçiş gibi yöntemleri uygulayarak sürdürülebilir enerji stratejilerini güçlendirir (Sun vd., 2019). Örneğin bir endüstriyel tesisin çatısına kurulan güneş panelleri ile elde edilen enerjinin kullanımı, küresel çapta enerji verimliliğini artırmak adına benimsenen bir uygulama olabilir. Bu stratejilerin uygulanması, sadece enerji maliyetlerini azaltmakla kalmayıp aynı zamanda karbon ayak izini minimize etme ve doğal kaynakların daha etkili bir şekilde kullanılması gibi çevresel faydalar sağlama potansiyeline sahiptir (Ding vd, 2021). Enerji verimliliği, endüstrinin sürdürülebilirlik yolculuğunda önemli bir adım olarak öne çıkarken (Mishra vd., 2020), uluslararası düzeyde bu konuda ortak çabalar ve standartlar belirlenmesi, endüstriyel sektörlerin sürdürülebilirlik alanında daha etkin bir rol oynamalarına olanak tanıyacaktır. 2.3.Atık Azaltma Endüstriyel süreçlerin sürdürülebilirliğini sağlamak ve çevre üzerindeki etkilerini azaltmak söz konusu olduğunda atık azaltma kavramı son derece önemlidir. İşletmeler, doğal kaynakları verimli bir şekilde kullanarak, çevre Paşa Gültaş \| 32 321 Paşa Gültaş kirliliğini azaltarak ve ekosistemlere verilen zararı en aza indirerek atıkları etkili bir şekilde yönetmeye çalışmaktadır (Pujara vd., 2019). Atık azaltma stratejileri sürdürülebilir üretim uygulamalarının çok önemli bir yönü olarak kabul edilmektedir. Başlangıçta, atık azaltma, üretim süreçlerinde kaynak kullanımını optimize etmeyi içerir. Bu, malzeme seçimi, tasarım ve üretim aşamalarında geri dönüştürülebilir ve yeniden kullanılabilir malzemelerin kullanımına öncelik verilmesini gerektirmektedir (Carvalho vd., 2018). İşletmeler, hammadde tüketimini en aza indirerek ve ürün tasarımında döngüsel ekonomi ilkelerini benimseyerek (Reuter vd., 2019) atık oluşumunu sınırlamaya yönelik ilk adımı atmaktadır (Wang vd., 2020). Malzemelerin geri dönüşümü ve yeniden kullanılması, kaynakların verimli kullanımını kolaylaştırdıkları için atık yönetiminde çok önemli bir rol oynar. İşletmeler geri dönüştürülebilir malzemeleri tanımlar ve bunları toplamak, işlemek ve üretim süreçlerine entegre etmek için stratejiler geliştirmektedir (Fisher ve Barron, 2019). Teknolojik gelişmeler, atıkların azaltılması için yeni fırsatlar sunmaktadır. Gelişmiş teknolojinin kullanılması ve süreç iyileştirmelerinin uygulanması, işletmelerin atıkları daha etkili bir şekilde yönetmesini sağlamaktadır (Litvinenko, 2020). Örneğin, akıllı sensörlerin üretim süreçlerine dahil edilmesi, atık oluşumunu izlemek ve en aza indirmek için yüksek bir görünürlük seviyesi oluşturmayı desteklemektedir (Astill vd., 2020). Çevresel etkilerin en aza indirilmesi, endüstriyel faaliyetlerin doğal ekosistemler üzerindeki olumsuz etkilerini azaltmada özellikle çok önemlidir. Bunu başarabilmek, endüstriyel tesislerin çevresel etkilerini izlemek ve değerlendirmek için çeşitli yöntemlerin uygulanmasını gerektirir (Gibb vd., 2019). Bir enerji üretim tesisinin çevresel etkisini en aza indirmek için karbon ayak izi izleme sistemlerinin kullanımı ve doğal habitatlara zarar vermemek için özel üretim tesislerinin tasarımı bunlara örnek olarak verilebilir. 3. GELECEĞİN SÜRDÜRÜLEBİLİR ÜRETİM TRENDLERİ İnovasyon ve sürdürülebilirlik alanındaki gelecek trendlerini öngörmek, küresel düzeyde çevresel ve sosyal sorumluluğa odaklanan yeni yaklaşımları anlamak ve gelecekteki sürdürülebilirlik çabalarını şekillendirmek açısından büyük bir öneme sahiptir. Bu alanlardaki gelecek trendleri, çeşitli faktörlerin etkileşimi sonucu ortaya çıkan dinamik bir süreci yansıtarak sürdürülebilirlik hedeflerine ulaşmak için işletmelerin ve toplumların nasıl evrileceğini belirlemektedir. 322 Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri Gelecekteki inovasyon trendleri arasında, yeşil teknolojilerin gelişimi ve benimsenmesi öne çıkmaktadır (Du ve Li, 2019). Temiz enerji, enerji depolama çözümleri, çevre dostu ulaşım araçları ve sürdürülebilir tarım teknolojileri gibi alanlarda gerçekleşecek inovasyonlar, endüstriyel sektörlerin çevresel etkilerini azaltma potansiyelini artıracak ve sürdürülebilirliği destekleyecektir (Khan vd., 2019). Sürdürülebilirlik açısından önemli bir trend, döngüsel ekonomi modellerinin yaygınlaşması olacaktır. Bu model, atıkların minimumda tutulması ve kaynakların sürekli olarak kullanılması prensibine dayanmaktadır (Hysa vd., 2020). İnovatif iş modelleri ve ürün tasarımları, kullanımdan sonra ürünlerin geri dönüştürülerek tekrar değerlendirilmesini sağlayacak ve doğal kaynakların daha etkili bir şekilde kullanılmasını teşvik edecektir (Ferasso, 2020). Gelecekte, sürdürülebilirlik odaklı finansal modellerin yükselmesi de beklenmektedir. Yeşil finans, çevre dostu projelere yatırım yapmayı amaçlayan finansal araçları içerir ve bu alanın büyümesi, sürdürülebilirlikle ilgili inovasyonların desteklenmesine ve sürdürülebilir projelerin finanse edilmesine yönelik bir eğilimi yansıtmaktadır (Süß vd., 2021). Bu trendlerin ötesinde, özellikle yapay zekâ, büyük veri analitiği ve nesnelerin interneti gibi yeni teknolojik gelişmelerin sürdürülebilirlik alanında nasıl kullanılabileceği konularına da odaklanılmıştır (Bibri, 2018; Andronie vd., 2021). Bu teknolojiler, enerji yönetimi, su tasarrufu, iklim değişikliği tahminleri gibi konularda daha etkili çözümler sunarak sürdürülebilirlik çabalarına katkı sağlayabilir (Vishwakarma vd., 2019). Sonuç olarak, gelecekteki inovasyon ve sürdürülebilirlik trendleri hem teknolojik gelişmelerin etkisiyle şekillenecek hem de küresel düzeyde çevresel ve sosyal sorumluluğa olan taleplerin artmasıyla yönlendirilecektir. Bu bağlamda, işletmelerin ve toplumların bu trendlere adaptasyonu, sürdürülebilir bir geleceğin inşasında kritik bir rol oynayacaktır. TARTIŞMA VE SONUÇ Gelecekte işletmelerin sürdürülebilirlik stratejilerini şekillendirmek, çeşitli faktörleri içeren karmaşık bir süreci gerektirir ve bu süreç, işletmelerin hem çevresel hem de sosyal sorumluluklarını etkin bir şekilde yerine getirmelerini hedefler. Bu bağlamda, işletmelerin gelecekte sürdürülebilirlik stratejilerini şekillendirebilmek için bazı temel unsurlar vardır. • İlk olarak, işletmelerin sürdürülebilirlik stratejilerini belirlerken, paydaş katılımı ve paydaşların beklentilerinin dikkate alınması önemlidir. Bu, müşterilerden tedarikçilere, yerel topluluklardan yatırımcılara kadar 323 Paşa Gültaş Paşa Gültaş geniş bir paydaş kitlesini içerir. Paydaş beklentilerinin anlaşılması ve bu beklentilere uygun stratejilerin oluşturulması, işletmelerin sürdürülebilirlik alanında daha etkili bir şekilde faaliyet göstermelerine katkı sağlar (Kaur ve Lodhia, 2018). • İkinci olarak, gelecekteki sürdürülebilirlik stratejilerini şekillendirmek için teknolojik inovasyonun önemli bir rol oynayacağı öngörülmektedir. Yenilenebilir enerji kaynakları, enerji verimli teknolojiler, sürdürülebilir üretim süreçleri ve yeşil teknolojiler, işletmelerin çevresel etkilerini azaltmalarına ve sürdürülebilirlik hedeflerine ulaşmalarına olanak tanır (Mousavi vd., 2019). Bu nedenle, işletmelerin teknolojik gelişmeleri takip ederek bu yenilikleri stratejik olarak entegre etmeleri kritik bir öneme sahiptir. • Üçüncü olarak, işletmelerin gelecekte sürdürülebilirlik stratejilerini şekillendirmelerinde, küresel ve yerel düzeydeki sürdürülebilirlik düzenlemelerine ve standartlarına uyum sağlamak kritik bir faktördür (Lambin vd., 2018). Bu düzenlemeler, işletmelerin çevresel performanslarını izlemelerini, raporlamalarını ve sürekli olarak iyileştirmelerini teşvik eder. Bu bağlamda, işletmelerin etiketlenme, sertifikasyon ve raporlama süreçlerini yönetmeleri önemlidir. • Son olarak, işletmelerin gelecekte sürdürülebilirlik stratejilerini belirlerken, döngüsel ekonomi prensiplerine odaklanmaları beklenmektedir (Suárez-Eiroa vd., 2019). Atık azaltımı, geri dönüşüm, ürün tasarımında sürdürülebilirlik ve tüketici bilinci gibi döngüsel ekonomi unsurları, işletmelerin kaynakları daha etkili bir şekilde kullanmalarını ve atıkları en aza indirmelerini sağlar. Tüm bu unsurların bir araya gelmesi, işletmelerin gelecekte sürdürülebilirlik stratejilerini şekillendirmelerini destekleyerek hem çevresel hem de ekonomik sürdürülebilirlik anlayışını güçlendirebilir. İşletmelerin bu stratejileri uygularken bilimsel yöntemlere dayalı analizler ve sürekli değerlendirmeler yapmaları, sürdürülebilirlik hedeflerine ulaşmalarında etkili olacaktır. Sonuç olarak, işletmelerin inovasyon ve sürdürülebilirlik alanında karşılaştıkları meydan okumalara karşı başarılı stratejiler geliştirmeleri hem kendi rekabet avantajlarını artırmalarına hem de toplumsal ve çevresel sorumluluklarına uygun bir şekilde hareket etmelerine olanak tanıyacaktır. Bu kitap bölümü, işletmelere, sürdürülebilirlik odaklı bir geleceği şekillendirmek için kullanabilecekleri değerli perspektifler ve uygulanabilir çözümler sunma amacı gütmüştür. Geleceğin iş dünyasında, inovasyon ve sürdürülebilirlik el ele yürüyerek işletmeleri daha dirençli, etik ve çevresel açıdan sorumlu hale getirecektir. 324 324 \| Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri KAYNAKÇA Adams, R., Jeanrenaud, S., Bessant, J., Denyer, D., & Overy, P. (2016). 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(2020). Circular eco- nomy innovation and environmental sustainability impact on economic growth: An integrated model for sustainable development. Sustainabi- lity, 12(12), 4831. Husgafvel, R., Pajunen, N., Virtanen, K., Paavola, I. L., Päällysaho, M., Inki- nen, V., ... & Ekroos, A. (2015). Social sustainability performance indi- cators–experiences from process industry. International Journal of Sustai- nable Engineering, 8(1), 14-25. Ilankoon, I. M. S. K., Ghorbani, Y., Chong, M. N., Herath, G., Moyo, T., & Petersen, J. (2018). E-waste in the international context–A review of tra- de flows, regulations, hazards, waste management strategies and techno- logies for value recovery. Waste management, 82, 258-275. Kaur, A., & Lodhia, S. (2018). Stakeholder engagement in sustainability ac- counting and reporting: A study of Australian local councils. Accounting, Auditing & Accountability Journal, 31(1), 338-368. Khan, N., Dilshad, S., Khalid, R., Kalair, A. R., & Abas, N. (2019). 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Journal of Cleaner Production, 256, 120268. 327 Paşa Gültaş Paşa Gültaş Schaltegger, S., Lüdeke-Freund, F., & Hansen, E. G. (2012). Business cases for sustainability: the role of business model innovation for corporate sustainability. International journal of innovation and sustainable devel- opment, 6(2), 95-119. Schaltegger, S., Lüdeke-Freund, F., & Hansen, E. G. (2012). Business cases for sustainability: the role of business model innovation for corporate sustainability. International journal of innovation and sustainable devel- opment, 6(2), 95-119. Seebode, D., Jeanrenaud, S., & Bessant, J. (2012). Managing innovation for sustainability. R&d Management, 42(3), 195-206. Mousavi, S., Bossink, B., & van Vliet, M. (2019). 328 \| Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri Zakari, A., Khan, I., Tan, D., Alvarado, R., & Dagar, V. (2022). Energy effi- ciency and sustainable development goals (SDGs). Energy, 239, 122365. Özet Kapitalist sistemin en önemli uluslararası finansal krizi olan Asya Krizi 1997 yılı Temmuz ayında başlamış ve öncelikle Tayland, Malezya, Endonezya ve Filipinler’de mali piyasaların zarar gördüğü bir bunalım yaşanmıştır. Bu kriz bütün dünyayı etkisi altına almıştır. Bu çalışmada ortaya çıkan dünya çapındaki krizin neden ve nasıl oluştuğuyla beraber bu oluşum diğer piyasaları nasıl etkiledi üzerinde durmuştur. Bu krizden kurtulmak için her ülke farklı yol ve yöntem izlemiştir. Krizden kurtulmak için izlenilen politikalar araştırılmıştır. KAYNAKÇA Microfoundations of com- panies’ dynamic capabilities for environmentally sustainable innovation: Case study insights from high‐tech innovation in science‐based compa- nies. Business Strategy and the Environment, 28(2), 366-387. Pujara, Y., Pathak, P., Sharma, A., & Govani, J. (2019). Review on Indian Mu- nicipal Solid Waste Management practices for reduction of environmental impacts to achieve sustainable development goals. Journal of environmen- tal management, 248, 109238. Reuter, M. A., van Schaik, A., Gutzmer, J., Bartie, N., & Abadías-Llamas, A. (2019). Challenges of the circular economy: a material, metallurgical, and product design perspective. Annual Review of Materials Research, 49, 253-274. Singh, S. K., Del Giudice, M., Chierici, R., & Graziano, D. (2020). Green in- novation and environmental performance: The role of green transforma- tional leadership and green human resource management. Technological forecasting and social change, 150, 119762. Song, W., & Yu, H. (2018). Green innovation strategy and green innovation: The roles of green creativity and green organizational identity. Corporate Social Responsibility and Environmental Management, 25(2), 135-150. Suárez-Eiroa, B., Fernández, E., Méndez-Martínez, G., & Soto-Oñate, D. (2019). Operational principles of circular economy for sustainable deve- lopment: Linking theory and practice. Journal of cleaner production, 214, 952-961. Sun, H., Edziah, B. K., Sun, C., & Kporsu, A. K. (2019). Institutional quality, green innovation and energy efficiency. Energy policy, 135, 111002. Süß, A., Höse, K., & Götze, U. (2021). Sustainability-oriented business model evaluation—a literature review. Sustainability, 13(19), 10908. Vishwakarma, S. K., Upadhyaya, P., Kumari, B., & Mishra, A. K. (2019). Smart energy efficient home automation system using IoT. In 2019 4th international conference on internet of things: Smart innovation and usages (IoT-SIU) (pp. 1-4). Ieee. Wang, F., Feng, L., Li, J., & Wang, L. (2020). Environmental regulation, tenure length of officials, and green innovation of enterprises. International Jour- nal of Environmental Research and Public Health, 17(7), 2284. 328 \| Yeşil İnovasyon ve Sürdürülebilir Üretim Stratejileri Zakari, A., Khan, I., Tan, D., Alvarado, R., & Dagar, V. (2022). Energy effi- ciency and sustainable development goals (SDGs). Energy, 239, 122365. Bölüm 18 1. GİRİŞ Ekonomik krizi Türk Dil Kurumu şöyle tanımlamaktadır; Bir ülkede veya ülkeler arasında, toplumun veya kuruluşların yaşamlarında görülen zorlu dönem, bunalım veya buhrandır. Kriz sözcüğünün karşılığı çöküntü olarak ifade edilmektedir. (Kaykusuz, 2014) Ekonomik kriz makro düzeyde devleti, mikro düzeyde şirketleri etkileyerek büyük sorunlar doğmasına neden olabilir. Krizlerde süre, organizasyonların krize karşı aldıkları önlemleri zamanında uygulamasına bağlıdır. (Aktan & Şen, 2001) Ekonomik Krizler reel sektör krizleri ve finansal krizler olarak ikiye ayrılmaktadır. Reel sektör krizleri; mal ve hizmet piyasasında meydana gelen kriz olarak tanımlanmaktadır. Reel sektör krizleri arasında yer alan enflasyon krizi mal ve hizmet piyasasındaki fiyatlarındaki artışın belli bir sınırın üzerinde oluşmasıyla meydana çıkmaktadır. Durgunluk krizi, fiyatlar genel seviyesindeki yükselişlerin ekonomide yatırımları teşvik edecek standartın altında olması durumunda reel sektör krizi yaşanmakta olduğu görülmektedir. Ayrıca işsizlik krizi de emek piyasasındaki işsizlik oranlarının 1 Öğr. Gör. https://doi.org/10.58830/ozgur.pub401.c1766 https://doi.org/10.58830/ozgur.pub401.c1766 329 330 Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar süregelen seviyenin üzerinde oluşması durumunda meydana gelmektedir. (Kibritçioğlu, 2001) Finansal krizler, kısa vadeli faiz oranları, varlık fiyatları, ödemelerin bozulması ve şirketlerin batması ile finansal kurumların iflas etmesi gibi göstergelerin tümünün hızlı, keskin bir şekilde bozulması şeklinde tanımlanmıştır. (Klein, 1947) Yirminci yüzyılın başlarında birçok finansal kriz bankacılık krizi gibi oluşmuştur. Borsa çöküşleri ve ekonomideki “balonların patlaması” ile döviz krizleri de finansal krizler arasında yer almaktadır. Bankacılık krizleri, para piyasası ve döviz piyasası krizleri, ödemeler dengesi krizleri, balonlar ve borsa krizleri genellikle finansal kriz olarak incelenmektedir. (Kaykusuz, 2014) Bu çalışmada kapitalist sistemin en önemli uluslararası finansal krizi olan Asya krizi incelenmektedir. 2 Temmuz 1997 yılında Tayland’ın para birimi olan Baht üzerinde spekülatif saldırılarda bulunulmuştur. Bunun üzerine Tayland hükümeti sabit döviz kuru rejimini terk etmek zorunda kaldığı görülmektedir ve bunun üzerine bahtı devalüe etmiştir. Tayland’ın ardından döviz kuru spekülasyonu bölgedeki diğer ülkelere yayıldı ve 1929 Büyük Burhan’ından sonra yaşanan en büyük ve en geniş alana yayılan uluslararası finansal kriz oldu. (Stiglitz, 2016) 1994 yılında Meksika’da başlayıp diğer Latin Ülkelerine de sıçramasıyla Tekila krizinden çok daha büyük bir kriz olmuştur. (İsa, 2009) 2. KAVRAMSAL ÇERÇEVE Asya krizinde kriz yaşanmadan önce makroekonomik temeller dengeli ve tutarlı olduğu gözlemlenmektedir. Kriz ülkelerinde makro ve mikro dengesizlikler olmaktaydı fakat gene bunlarda kriz ortamı doğuracak kadar şiddetli değildi. Krize giren beş Asya ülkesi arasında makro ve mikro temelleri en sağlam durumda olan ülke Endonezya ekonomisiydi. Fakat krizden en derin yara alan ülkede gene Endonezya olmuştur. (Cengiz, 2010) 1993 yılında Dünya Bankası’nın yayınladığı raporda “Doğu Asya Mucizesi” olarak adlandırdığı ve övgü ile bahsedilmiş olan kalkınma modeli şöyledir; Japonya’nın yirminci yüzyılın son çeyreğinde meydana gelen sanayileşme ile Asya’da bulunan Malezya, Endonezya ve Tayland ile Asya Kaplanları olarak bilinen Hong Kong, Singapur, Tayvan ve Güney Kore’yi içerisine almaktadır. Bölge ekonomileri tarıma dayalı olan bu ülkeler çok kısa zamanda teknolojik ve sosyoekonomik değişimi başarı ile sağladılar. 1970’liyıllardan itibaren hızla büyüyen ekonomileri “mucize” olarak ifade edilecek bir haklılığı ortaya koymaktadır. (Öksüz, 2001) 1970’den itibaren bütün ekonomik kalkınma içerisinde önemli yol aldıkları gözlemlenmiştir. Ayrıca bazı literatürlerde ise Berna Ağbulut \| 331 Berna Ağbulut \| 331 331 bu bölgelerde gerek toplumsal gerekse kültürel ve ayrıca da siyasi yapıların farklı olmasının yanı sıra sol kesimi Asya Kriz’ini, kapitalist sistemin temeli olan 1970’li yıllardaki aşırı üretimin olmasına bağlamış oldukları gözlemlenmiştir. (Sungur, 1999) 1970’li yıllarda başlayıp 1980’lerde hızla ilerleyen finansal liberalleşme ve uyum süreci 1990’lı yıllardan sonraki dönemde küreselleşmenin etkisi ile dünya çapında hızla ilerlemiş ve uyum sağlandığı görülmüştür. Finansal entegrasyon yararı olduğu gibi gelişmekte olan ülkeler açısından finansal krizlerin yaşanmasına ve buna bağlı olarak ta işsizlik, yoksulluk gibi durumların doğmasına neden olmaktadır. Bu krizlerden biriside 1997 Güney Doğu Asya Krizi’dir. (Turgut, 2006) Asya’da geniş çaplı bir finansal kriz olması beklenen bir fikir değildi. 2008 yılı Nobel Ekonomi Ödül’ünü kazanan Krugman kriz öncesi dönemde 1994 yılında bölgedeki hızlı gelişme ve ekonomik kalkınmayı eleştirmiştir. Ona göre Doğu Asya ülkelerinin kalkınmasına neden olan etkenler doğal kaynakların yer değiştirebilir özellikte olması, sermaye yapısı, emek gücü ve daha önemle bakılan bu ülkelerdeki yüksek tasarruf oranları ve bu tasarrufların yatırıma dönüştürülmesinin devamlı ve sürekli olarak sağlanamayacağının altını çizmiştir. Krugman’a göre doğal kaynakların, sermayenin ve emeğin sınırı vardır. Bu sınır büyüme hızının düşmesine neden olacağını düşündüğünü belirtmiştir. Fakat kriz olduktan sonra bu denli büyük bir kriz olacağını da düşünmediğini ifade etmektedir. (Krugman P.1998) Bütün krizlerde olduğu gibi Asya Kriz ’inde de tek bir nedeni yoktur krizin, birden fazla etkileyicisi vardır. Krizden önceki dönemde özellikle de Tayland, Güney Kore ve Endonezya ekonomilerinin dışarıdan gelecek şoklara karşı savunmasız oldukları gözlemlenmiştir. 2. KAVRAMSAL ÇERÇEVE Yüksek getirili kısa vadeli küresel sermaye akışını savunan makroekonomik politikalar, yeni liberalleştirilmiş ve gerekli düzenlemeler yapılmamış olan yurtiçi finansal piyasalar ve artan siyasi belirsizlik olarak sorunları sıralamışlardır. Ayrıca kriz nedeni olarak şunları dile getirmişlerdir (Schmukler, vd., 2001). I. Kısa vadeli küresel sermaye ile finanse edilen ve gittikçe genişleyen cari açık, II. Finansal piyasalarda istenilen düzeyde düzenlemelerin yapılmaması sonucu, bankaların ve finansal kurumların yurtdışından dolar ile kısa vadeli borçlanması ve yurtiçinde kendi para birimleri ile uzun vadeli kredi vermeleri sonucu ortaya çıkan kur ve vade uyumsuzluğu riskleri, 332 \| Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar III. Firmaların gerek ülke içerisinde, gerekse ülke dışından aldıkları kredileri uzun vadede gelir getiren verimsiz alanlarda kullanmaları, III. Firmaların gerek ülke içerisinde, gerekse ülke dışından aldıkları kredileri uzun vadede gelir getiren verimsiz alanlarda kullanmaları, III. Firmaların gerek ülke içerisinde, gerekse ülke dışından aldıkları kredileri uzun vadede gelir getiren verimsiz alanlarda kullanmaları, IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. Ayrıca bankacılık sektörü ile şirketlerin aldıkları yurtdışı kredilerin ve kısa vadeli portföy yatırımlarının Asya Kriz ’inin önemli nedeni olduğu görülmektedir. Bölge ülkelerindeki bankacılık dâhil finansal sektörlerde ki denetimlerin yetersiz kalması ve verilen kredilerin doğru yerlerde kullanılmamış olması da krize neden olan bir durumdur (Chan-Lau & Chen, 1998). Krizlerin ayrıştırılmasında birçok yöntem kullanılmaktadır. Bunlar krizin sonucuna göre veya krizin nedenine göre uygulanmaktadır. Finansal krizlerin nedenleri birinci ve ikinci nesil finansal kriz yöntemleri olmak üzere iki yaklaşımda anlatılmıştır. Son zamanlarda üçüncü nesil kriz yöntemi de eklenmiştir. Birinci nesil (kanonik) modelleri, krizin oluşmasına neden olan esas makro ekonomi faktörlerin önemliliği üzerinde durmakta ve hiç beklenmedik anda spekülatif hareketle başlayan para/döviz krizlerine sebep olanları, sabit döviz kurunun devamlılığı ile tutarsız iktisadi politika uygulamalarını etken olarak görmektedir. Bu tutarsız politikalara verilebilecek en dikkat çeken örnek ise; bütçede meydana gelen açıkları para basarak finanse etmeye neden olan genişleyici para politikaları ile sabit/yarı sabit kur rejiminin ortak uygulanmasıdır. (Çakmak, 2007) Birinci nesil kriz modellerinin monetarist kesimin oluşturduğu düşünülmektedir. Monetaristlerin, Friedman ve Schwarts çalışmaları ile başlamışlar ve finansal krizleri bankacılık kriziyle ilişkili olduğunun önemi üzerinde durmuşlardır. Monetaristler, varlıkların fiyatlarındaki azalışların iflaslarda artışa neden olmasına rağmen, panik ortamının oluşmadığı ve para arzının net bir düşüşle sonuçlanan olayları, gerçek finansal krizler olarak görmezler. Bu gerçek olmayan krize de devletin müdahale etmesini gereksiz olduğuna ve hatta enflasyona neden olacağı için zarar getireceği fikrine dikkat çekmişlerdir. (Oktar & Dalyancı, 2010) Birinci nesil modellerde önemle üzerinde durulması gereken iki önemli husus vardır: I. Ülke içerisinde uygulanan ekonomi politikaları ve döviz kurundan kaynaklanan epeyce basite indirgenmiş olan bu model çoğunlukla döviz kuru sistemlerinde çözümlenmesi zor durumlarda da bir örnek olarak gösterilmektedir. Berna Ağbulut \| 333 333 Berna Ağbulut II. Model sadece tek olarak yatırımcıların ve piyasaya şekil verenlerin davranışlarının, kayıp oluşturması hızlı ve birden yaşanan döviz krizinin tek nedeni olmayacağının önemi üzerinde durmaktadır. Birinci nesil model ile ilgili doğan eksiklikler; ikinci ve üçüncü nesil modellere ihtiyaç doğmasına neden olmaktadır. Birinci neslin temel özelliklerini şöyle sıralamak mümkündür (Krugman P., 1979): I. Yerli paranın döviz kuruna bağlı olması I. Yerli paranın döviz kuruna bağlı olması II. Yerli paranın piyasasında kur aracılığı ile dönem dönem dengeye gelmesi III. Sadece bir ticari mal üreten küçük bir ülke varsayımı doğrultusunda hedef gösterilen malın fiyatının dünya piyasasında belirlenmesi durumu üzerine satın alma paritesine göre döviz kurunun ulusal fiyat seviyesini belirlediğinden bahsedilebilir. IV. Ülke fiyatlarının ve ücretlerinin tam esnek, üretiminin tam istihdam seviyesinde olması V. IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. Ödemeler dengesi yerine cari denge kullanılması ve cari dengenin de üretim ve harcama arasındaki farktan kaynaklanması fikri varsayılmıştır. VI. Varlık piyasası ile ilgili temel görüşler yatrım yapan kişilerin tek yerli ve yabancı para ile olan seçimlerin dahil olabileceği, her iki para için faizin hiç olmadığı ve yurt içinde yaşayanların reel servetlerinin yerli ve yapancı paraların reel toplamına eşit olduğu VII. Yabancı kişilerin yerli para tutmayı tercih etmediği ve portföy dengesi servetin belli bir oranı olarak değerlendirilmiştir. Bu oran beklenen enflasyon oranı ile belirlenmektedir. Birinci nesil modeller devletin kontrolsüz makroekonomik politikaları uygulaması, ikinci modellerin de hükümetin sabit kur sistemini devamı konusunda kredibilite yetersizliğini ön plana çıkartmış olup, Doğu Asya kriziyle beraber hükümet içerisinde farklı bir alanda başarısızlık doğması üçüncü modellemenin oluşmasına yol açmıştır. Finansal piyasalar açısından gerekli kontrol ve yönetiminden eksik olmaları krizde önemli bir yer tutmuştur. (Aghion vd., 2009) 1997 yılında başlamış olan Doğu Asya Krizi Tayland, Endonezya, Malezya, Filipinliler ve Kore’den oluşmaktadır. Bu ülkelerin yaşamış oldukları krizde farklı etkileri olmuş olsa da ortak noktalarının da olduğu görülmektedir. Üçüncü nesil modelin gelişmesinde öncülük eden kriz olan 1997 Doğu Asya krizi üç önemli durum üzerinde durmaktadır. Bunlar; ahlaki çöküntünün neden olduğu dış borç, Diamond- 334 Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar Dybving’in bankacılı1997 yılında başlamış olan Doğu Asya Krizi Tayland, Endonezya, Malezya, Filipinliler ve Kore’den oluşmaktadır. Bu ülkelerin yaşamış oldukları krizde farklı etkileri olmuş olsa da ortak noktalarının da olduğu görülmektedir. Üçüncü nesil modelin gelişmesinde öncülük eden kriz olan 1997 Doğu Asya krizi üç önemli durum üzerinde durmaktadır. Bunlar; ahlaki çöküntünün neden olduğu dış borç, Diamond-Dybving’in bankacılık sistemine yönelik hücum modelinin açık ekonomilerde oluşturduğu durum ve son olarak da yerli paranın değerinin düşmesi sonucunda bilançolar üzerinde oluşturmuş olduğu etkidir. (Krugman P. , Balance Sheets, the Transfer Problem, and Financial Crises, 2009) Diamond-Dybving’un dışa açık modelinde bankacılık sisteminde üç farklı döviz kuru göz önüne alınmıştır bunlar para kurulu, sabit kur ve esnek kurdur. Merkez bankasının kredi politikasını göz önünde bulundurarak banka hücumlarını gerçekleştirme durumunu incelemiştir ve bunun sonucunda esnek kur sisteminde banka hücumlarının olası olduğu ve sabit kur sisteminde, para kuruluna göre bankanın hücumlarına ve döviz krizlerine daha uygun ortam yaratıldığı görüşüne varılmıştır (İsa, 2009). Asya Krizi’nde IMF tarafından desteklenen program uygulamaları ile iç talebin yatırım tarafıyla birlikte tüketim, de bitirmiştir. Asya ülkelerine önemli düzeyde resmi dış kaynak girişi taahhüt edildiği için bitişin bu kadar yüksek düzeyde olması beklenmemişti. IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. Bu duruma karşın bu ülkelerde hem yabancı fon çıkışının sürmesi, hem de bankacılık kısmının düşmesiyle finansman kaynakları çok geriledi. Finansman darboğazı bir kredi çöküntüsüne dönüştü. (Akyüz, 2000) Diamond-Dybving’un dışa açık modelinde bankacılık sisteminde üç farklı döviz kuru göz önüne alınmıştır bunlar para kurulu, sabit kur ve esnek kurdur. Merkez bankasının kredi politikasını göz önünde bulundurarak banka hücumlarını gerçekleştirme durumunu incelemiştir ve bunun sonucunda esnek kur sisteminde banka hücumlarının olası olduğu ve sabit kur sisteminde, para kuruluna göre bankanın hücumlarına ve döviz krizlerine daha uygun ortam yaratıldığı görüşüne varılmıştır (İsa, 2009). Asya Krizinde IMF’nin desteklediği programlar ile iç piyasa talebin yatırım tarafı dâhil edilerek tüketim de düşmüştür. Asya ülkelerine önemli düzeyde resmi dış kaynak girişi taahhüt edildiği için düşüşün bu kadar ileri düzeyde olması beklenmemişti. Buna karşın bu ülkelerde hem yabancı fon çıkışının sürdürülmesi, hem de bankacılık kesiminin düşmesi finansman kaynakları çok geriledi. Finansman darboğazı bir kredi düşüşüne neden oldu. (Akyüz, 2000) 335 Berna Ağbulut İkinci nesil modelleri: Kronik modelde oluşan tamamlanmamış tarafları bulunan en iyi hükümet politikalarının mekanik tanımlandığını söylemektedir. Esasında uygulanan politika ihtimallerinin aralığı fazlasıyla geniştir. Merkez bankalarının elinde öncelikle yurt içi para arzını daraltmak ve döviz kurunu koruyucu çeşitli argümanlar bulunmaktadır. İkinci nesil modellerde üç ana bileşen vardır; I. Devletin sabit döviz kurunu kullanmak istememelerinin bir nedeni olması II. Devlet tarafından sabit kur rejimini korumak istemelerinin nedeni olmalı III. Krize neden olan kısır döngünün oluşması için sabit kuru korumanın maliyeti ciddi oranda arttırılmalıdır ve böylece insanlar sabit kurun sona ereceği fikrine inanmalıdırlar. Devlet tarafından uygulanan politikalardaki uyumsuzluğun piyasa spekülatörlerinin kazançlarını arttırmak amacıyla izlenme durumunu yaratmaktadır. Bu gözlem spekülatörlerin kriz ortamı yaratmasına neden olmaktadır (Krugman P., 1979). İkinci nesil krizlerde meydana gelen hareketlenmenin zamanlaması önemlidir. Piyasada meydana gelen herhangi bir bilgiye piyasa oyuncularının inanması ve bu bilginin etkili bilgi haline gelmesi durumuna güneş lekesi tanımlaması yapılmaktadır. (Fisscher & Blanchard, 1990) İkinci nesil modeller hükümet ve spekülatörler ile karşılıklı etkileşimi ön planda tutmaktadır. Birinci nesil modellerden farklı olarak hükümetin pasif rolü aktif bir hal almaktadır. Hükümetin uygulayacağı ekonomi politikaları da piyasada oluşacak beklentiye göre hareket etmektedir. (İsa, 2009) İkinci nesil modellere göre finansal krizler kendi kendine etkili olabilen beklentilerin sonucu olarak meydana gelmiştir. Bu krizler tahmine açık olmayan ve bulaşıcı etkisi yüksek olan kriz olarak düşünülür ve kabul edilir. Yatırım yapan kişilerin döviz kruu paritesinin artmasına izin verme durumundan şüphelenmeye başladıklarında, yatrımların getiri oranı artar. Bununla beraber faiz oranları üzerinde ki baskınında artmasıyla hükümet sabit döviz kurunu bitirme durumuna gelir. IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. (Krugman P. , 2009) Asya mucizesi olarak ifade edilen ülkelerin Asya krizi yaşaması sonucunda birinci ve ikinci nesil modellerle çözümlenememesinin nedenini anlayabilmek için krizin merkezinde olan ülkelerin finansal yapılarını bilmek gerekmektedir. Birinci ve ikinci modellerde önemli etkileri olan devlette finansal sistemin kriz yaratması da önemli bir faktör olarak yer almaktadır. 336 Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar Asya krizini açıklamaya veya krizden kaçınmaya birçok farklı makro iktisat yaklaşımları vardır. Bunlardan ilki IMF’nin bu anlamda yapmış olduğu görüş ve önerilerdir. İMF Asya Krizinde daraltıcı politikalar izleyerek iç talebin kısılmasını, dış dengeyi kurmayı ve kur artışını belli bir düzeyde tutmaya yönelik yaklaşımlarda bulunmuştur. Ayrıca büyüme dikkate alınmamıştır. İMF’nin Asya krizinden çıkış politikalarının krizi durdurmak yerine aksine daha derin ve çıkılması zor bir noktaya geldiği düşüncesi P. Krugman, J. Sachs ve J. Stiglitz gibi iktisatçıların birleşmesini sağlayarak UNCTAD’ı içerisine alan bir yaklaşımın gelişmesine neden olmuştur. (Uygur, 2001) Asya krizini açıklamaya veya krizden kaçınmaya birçok farklı makro iktisat yaklaşımları vardır. Bunlardan ilki IMF’nin bu anlamda yapmış olduğu görüş ve önerilerdir. İMF Asya Krizinde daraltıcı politikalar izleyerek iç talebin kısılmasını, dış dengeyi kurmayı ve kur artışını belli bir düzeyde tutmaya yönelik yaklaşımlarda bulunmuştur. Ayrıca büyüme dikkate alınmamıştır. İMF’nin Asya krizinden çıkış politikalarının krizi durdurmak yerine aksine daha derin ve çıkılması zor bir noktaya geldiği düşüncesi P. Krugman, J. Sachs ve J. Stiglitz gibi iktisatçıların birleşmesini sağlayarak UNCTAD’ı içerisine alan bir yaklaşımın gelişmesine neden olmuştur. (Uygur, 2001) Asya Krizinde IMF destekli programların uygulanması ile iç talebin yatırım kısmıyla beraber tüketim de çökmüştür. Asya ülkelerine önemli seviyede resmi dış kaynak girişi taahhüt edildiği için çöküntünün bu derece ileri seviyede olması beklenmemişti. Buna rağmen bu ülkelerde hem yabancı fon çıkışının devam etmesi, hem de bankacılık kesiminin çökmesiyle finansman kaynakları çok geriledi. Finansman darboğazı bir kredi çöküntüsüne dönüştü. (Akyüz, 2000) 1997 yılında Doğu Asya Krizinin yaşanması sonucu döviz krizleri ve finansal krizleri açıklayan yeni bir modelin ortaya çıkmasına ihtiyaç doğurmuştur. Doğu Asya Krizi bütçe açıklarını finans eden yöntem olan sabit kur sistemindeki dengesizlikten kaynaklandığını ifade eden birinci nesil model ile açıklanamamıştır. Ayrıca makro ekonomik büyüklüklerin kriz oluşturacak boyutta olmaması spekülatörlerin devletin sabit kur sistemini devam ettiremeyeceği açısından oluşan beklentilerin sebebi olan ikinci nesil model Doğu Asya Krizini açıklaymamıştır. Bu sebebeple Doğu Asya Krizini açıklamak için üçüncü nesil model geliştirilmiştir. (İsa, 2009) Üçüncü nesil modeleller farklı üç temel konu üzerinde durmuştur bunlardan ilki ahlaki çöküntünü oluşturduğu ve bunun neden olduğu dış borçlanmadır. IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. Daha sonra Diamond – Dyvbing’in banka sektörüne dair hücum modelindeki açık ekonomide meyf-dana gelen durum ve son olarak da yerli para değer kaybedince bunun bilançolara yarattığı etkidir. (Krugman P., 2001) Asya krizi sırasında Endonezya, Kore ve Tayland sıkı maliye ve para politikalarına, faize ve büyümedeki hızlı gerileme olmasına karşılık döviz kurunda önemli artışlar olduğu izlenmiştir. Yaşanan 1997 Asya krizi sırasında ki kur artışları Tayland’da 1997 Ağustos ayında, Endonezya’da 1997 Kasım ayında, Kore’de 1997 Aralık ayında yürürlüğe firen İMF politikalarıyla hızlanmış aynı durum Brezilya’da da yaşanmıştır. Bu gelişmeler faiz ile kur artışı arasında ters ilişki olduğunu söyleyen IMF’nin yaklaşımıyla uyuşmamış olduğu gözlemlenmiştir. Yoğun eleştiriler alan IMF’nin bu yaklaşımına karşılık Berna Ağbulut \| 337 Berna Ağbulut \| 337 Berna Ağbulut \| 337 Krugman (1999), Radelet and Sach (1998), Furman and Stiglitz (1988) ile yeni bir yaklaşım fikrinin doğmasına neden oldukları görülmektedir. Bu da krizin yaşandığı durumda maliye ve para politikalarının esnek bırakılmasıyla beraber reel faizinde düşürülmesi gerektiği görüşüne değinilmiştir. Krizin sona ermesiyle meydana gelen belirsizliğin zaten daralma eğilimi içerisinde olan bir ekonomide bunun yanında paranın sıkılması ve faiz oranlarının artmasıyla da krizin iyice derinleşeceği fikrine varmışlardır. (Uygur, 2001) Krugman (1999), Radelet and Sach (1998), Furman and Stiglitz (1988) ile yeni bir yaklaşım fikrinin doğmasına neden oldukları görülmektedir. Bu da krizin yaşandığı durumda maliye ve para politikalarının esnek bırakılmasıyla beraber reel faizinde düşürülmesi gerektiği görüşüne değinilmiştir. Krizin sona ermesiyle meydana gelen belirsizliğin zaten daralma eğilimi içerisinde olan bir ekonomide bunun yanında paranın sıkılması ve faiz oranlarının artmasıyla da krizin iyice derinleşeceği fikrine varmışlardır. (Uygur, 2001) Bilindiği gibi dünya ticareti Japonya dışında Asya ülkeleri tarafından gerçekleştirilmektedir. Finansal krizin neden olduğu devalüasyon ülkelerin rekabet gücünün artması yönünde sonuçlar doğurmaktadır. Bu noktada uluslararası fonların, ekonomik gücü yüksek ülkelerin borçlanma enstrümanlarına yatırması olası bir durumdur. Finansal piyasalar küreselleşmeyle beraber oluşan günümüz piyasaları arasında simetrik risk oluşumuyla kriz bütün dünya ticaretinde daraltıcı etkisi olduğu görülmektedir. Başlangıçta bölge krizi gibi görünse de Hong Kong Borsası ve onu takiben ABD ve diğer gelişmiş ülkeler borsalarındaki büyük düşüşler bir anda dünya krizine dönüşmüştür. Bunun sonucunda uluslararası yatırımcıların portföy oluşturma sürecindeki yönelmelerinde keskin değişiklikler olmuştur. Bu yatırımcılar hiç olmazsa piyasalarsa yeni bir denge meydana gelene kadar gelişmekte olan ülkeler ve hisse senedi borsaları gibi daha riskli piyasalardan ayrılarak gelişmiş ülkelerin hazine bonoları, Alman Markı ve İsviçre Frangı gibi riski çok yüksek olmayan enstrümanları talep etmişlerdir. Uluslararası yatırımcılar gelişmekte olan piyasalardan çıkarken ülke ayrımı yapmamışlardır. IV. Özelliklede Endonezya’da oluşan siyasi belirsizlik. Bu durumun oluşmuş olmasına rağmen piyasadan ayrılmaları ölçü ve yarattığı etki açısından farklılıklar göstermiştir. Normal dönemlerde ki oluşumların tersine daha iyi ve güçlü performans gösteren piyasalar, yabancıların etkinliğinin daha düşük olduğu veya birden bire çıkılması kolay olmayacak derinlikte olan piyasalardır. Örneğin Malezya, Tayland, Hong Kong ve Brezilya piyasalarında büyük oranlı düşüşler olmuşken; Hindistan, Pakistan, Türkiye, Rusya ve Afrika piyasaları sınırlı düşüş olduğu gözlemlenmektedir. (Özdemir, 2000) Krizin tüm dünyayı etkiler hale dönüşmesinde her ne kadar Hong Kong piyasaları etkili gibi düşünülse de dünyada ki piyasaları esas etkilemiş olan Japon ekonomisinin içinde olduğu değişimdir. Krizin dünya etkisi önemli seviyelere yükselmiştir. Bu yükseliş Japonya ile adım atacakları uluslararası ticari ilişkilerin temel alındığı söylenebilir. Japon, ihracatının büyük kısmı Asya Kaplan’larına yaptığı gerçeği, ticaret açısından en olumsuz yönde etkileneceğinin göstergesi olmuştur. (Hendersen, 1999) 338 Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar Japon ekonomisindeki değişim dünya ekonomilerinin ilerleyen süreçlerinde içinden çıkılmaz bir durum oluşturmaktadır. Japonya’da meydana gelecek olan bir kriz bütün dünyayı etkisi altına alınacağı bilinmektedir. Krizi atlatıp hemen büyüme sürecine girildiğinde düşük olan faiz oranlarının yükseldiğini ve dış piyasalarda bulunan Japon fonlarının Japonya’ya geri dönmesi amaçlanarak piyasada çıkmalar olacaktır. Bölge de bulunan ülkelerin birden fazla alanda rakibi olan Çin’in sonraki gidişatında önemli görev üstlenen ülke olma özelliğini ortaya koymaktadır. Çin’de ki yavaşlayan büyüme hızı ve yüksek olan işsizlik sorunun mevcudiyeti bazı kaygılanmalara neden olmuştur. Bu durum “Küresel Deflasyon” olarak ifade edilen bir durumun yaşanması fikrine ulaştırmıştır. Bu durumda doğacak kayıpları önlemek için şu önlemler alınmaktadır. (Özdemir, 2000) I. Karşılaşılan spekülatif olaylarda bölge ülkelerinin verdiği ilk tepki fiyatı düşük olan ithal mallara yönelimi azaltmak için engel koymak ve ihracat yapılan malların rekabetini arttırmak amacıyla devalüasyon yapmak. II. Krizle beraber doğan bir takım sonuçlar, krizin başlamasından itibaren üç, dört yıl süreyle durgunluğun hissedileceği gözükmektedir. Bu durgunluğun ortaya çıkması uluslararası ticaretten ziyade çok uluslu firmaların çalışma gücüyle olacağı görülmektedir. III. Üretim fazlalığı öncesinde görülmemiş seviyelere çıkmıştır. Dünya otomobil sektörünün dünya ihracatında yüksek oranlı kapasitesi olduğu görülmektedir. Japon şirketlerin Asya’da kurdukları üretim kapasiteleri Fransa endüstrisinin üç katına ulaştığı görülmüştür. SONUÇ Sermaye hareketinin çok olduğu ülkelerde reel ekonomik değişimler Asya Krizinin doğmasına sebep olmuştur. Sermaye akımının oluşma nedenleri arasında gösterilen gelişmekte olan ülkelerdeki gayrimenkul ve hisse senedi fiyatlarının çok yüksek şekilde yükselme yaşamasıdır. Asya Krizi döneminde etkisini gösteren bu gelişme bazı piyasalarda malların değerinin ani ve hızlı yükselmesi nedeni olmuştur. Bu etkinin ani bir şekilde ortadan kalkması ise diğer bazı piyasalarda makro gelişmelerle de birleşince ülkelerin bazılarında finansal krizler meydana getirmektedir. (Özdemir, 2000) Asya ülkelerinde meydana gelen finansal panik kredi çöküntüsünü meydana getirmiş ve reel ekonomiyi negatif yönde etkilemiştir. Türkiye’de bu tür panik yaratacak daraltıcı durumlara ortam hazırlamamak adına hem tasarruf teminatının devam ettiği hem de bankaların kapatma kararı 339 Berna Ağbulut \| 33 vermeyecekleri açıklanmıştır. Bu yaklaşımın nedeni ise parasallaşma olarak ifade edilmekte olduğu görülmüştür. (Uygur, 2001) Asya Krizi, kamunun kriz döneminde borçlarını ödeyebilmenin önemini vurgulamıştır. Kore, Endonezya, Malezya ve Tayland gibi Doğu Asya ülkeleri Latin Amerika ülkesi olan Meksika’nın 1994’de ekonomik krizde meydana gelen olaylar ile benzerlik gösteren finansal problemler oluşmasına neden olduğu gözlemlenmiştir. Burada ki temel nedenin üst düzeyde gözlemlenen kısa vadeli sorumlulukları ve banka kredileri ile bankaların borçlarının maddi örtüşmemesidir. Banka alacakları, değeri düşen yerli paradan, borçları ise geçmişten daha değerli olan yabancı para cinsinden olması olarak ifade edilmiş olduğu görülmektedir. (Cole & Kehoe, 1998) 1997 yılında başlamış olan Doğu Asya Krizi Tayland, Endonezya, Malezya, Filipinliler ve Kore’den oluşmaktadır. Bu ülkelerin yaşamış oldukları krizde farklı etkileri olmuş olsa da ortak noktalarının da olduğu görülmektedir. Üçüncü nesil modelin gelişmesinde öncülük eden kriz olan 1997 Doğu Asya krizi üç önemli durum üzerinde durmaktadır. Bunlar; ahlaki çöküntünün neden olduğu dış borç, Diamond-Dybving’in bankacılık sistemine yönelik hücum modelinin açık ekonomilerde oluşturduğu durum ve son olarak da yerli paranın değerinin düşmesi sonucunda bilançolar üzerinde oluşturmuş olduğu etkidir. (Krugman P. , Balance Sheets, the Transfer Problem, and Financial Crises, 2009) Diamond-Dybving’un dışa açık modelinde bankacılık sisteminde üç farklı döviz kuru göz önüne alınmıştır bunlar para kurulu, sabit kur ve esnek kurdur. Merkez bankasının kredi politikasını göz önünde bulundurarak banka hücumlarını gerçekleştirme durumunu incelemiştir ve bunun sonucunda esnek kur sisteminde banka hücumlarının olası olduğu ve sabit kur sisteminde, para kuruluna göre bankanın hücumlarına ve döviz krizlerine daha uygun ortam yaratıldığı görüşüne varılmıştır (İsa, 2009). Asya Krizinde IMF destekli programların uygulanması ile iç talebin yatırım kısmıyla beraber tüketim de çökmüştür. Asya ülkelerine önemli seviyede resmi dış kaynak girişi taahhüt edildiği için çöküntünün bu derece ileri seviyede olması beklenmemişti. SONUÇ Buna rağmen bu ülkelerde hem yabancı fon çıkışının devam etmesi, hem de bankacılık kesiminin çökmesiyle finansman kaynakları çok geriledi. Finansman darboğazı bir kredi çöküntüsüne dönüştü. (Akyüz, 2000) 340 \| Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar KAYNAKÇA Aghion, P., Bacchetta, A., & Banerjee, A. (2009, Mart 5). Currency Crises And Monetary Policy İn An Economy With Credit Constraints. Https://Cepr. Org: Http://Www.Cepr.Org/Meets/Wkcn/1/1458/Papers/Bacchetta.Pdf Adresinden Alındı Aktan, C. C., & Şen, H. (2001). Ekonomik Kriz: Nedenleri Ve Etkileri. Yeni Türkiye Dergisi, 1225-1230. Akyüz, Y. (2000). Causes And Sources Of The Asian Financial Crisis. Symposium On Economic And Financial . Bangkok: UNCTAD X. Cengiz, S. (2010). Uluslararası Finansal Panik Yaklaşımı Ve 1997/8 Asya Krizi. Selçuk Üniversitesi Sosyal Bilimler Ensitüsü Dergisi, 173-181. Chan-Lau, J. A., & Chen, Z. (1998). Financial Crisis And Credit Crunch As A Result Of Inefficient Financial Intermediation—With Reference To The Asi- an Financial Crisis. INTERNATIONAL Monetary FUND. Cole, H. L., & Kehoe, T. J. (1998). Self-Fulfilling Debt Crises. Minneapolis: Fe- daral Reserve Bank Of Minneapolis Research Departmenr Staff Report 211. Çakmak, U. (2007). Kriz Modelleri Çerçevesinde Türkiye 2001 Finansal Krizi- nin Değerlendirilmesi. Gazi Üniversitesi İktisadi Ve İdari Bilimler Fakültesi Dergisi , 81-101. Fisscher, S., & Blanchard, O. J. (1990). Lectures On Macroeconomics. Cambridge (4. B.). The MIT Press. Hendersen, C. (1999). Asya Krizi Ve Sonrası “Asya’nın Çöküşü”. (M. Güvenç, Çev.) İstanbul: Alfa Yayınları. İsa, D. (2009). Türkiye’de Yaşanan 1994 Ve 2001 Ekonomik Krizlerinin Analizi. İstanbul: Marmara Üniversitesi Sosyal Bilimler Ensitüsü. Kaykusuz, M. (2014). Gemişten Günümüze Finansal Krizler. Bursa: Ekin Yayınevi. Kibritçioğlu, A. (2001). Türkiye’de Ekonomik Krizler Ve Hükümetler, 1969- 2001. Yeni Türkiye Dergisi, 171-190. Klein, L. R. (1947). The Keynesian Revolution. New York: Macmillan. Krugman, P. (1979). A Model Of Balance Of Payments Crises. Journal Of Mo- ney, Credit, And Banking, S. 311-325. Krugman, P. (1998, Ocak). WHAT HAPPENED TO ASIA? Https://Web.Mit. Edu/: Https://Web.Mit.Edu/Krugman/Www/DISINTER.Html Adre- sinden Alındı Krugman, P. (2001). Bunalım Ekonomisinin Geri Dönüşü (2. B.). (N. Domaniç, Çev.) İstanbul: Literatür Yayınları. Krugman, P. (2009, Eylül 07). Balance Sheets, The Transfer Problem, And Fi- nancial Crises. Https://Web.Mit.Edu/: Http://Web.Mit.Edu/Krugman/ Www/FLOOD.Pdf Adresinden Alındı Berna Ağbulut \| 341 341 Krugman, P. (2009, 09 7). Balance Sheets, The Transfer Problem, And Financial Crises. /Web.Mit.Edu: Http://Web.Mit.Edu/Krugman/Www/FLOOD. Pdf Adresinden Alındı Oktar, S., & Dalyancı, L. (2010). Finansal Kriz Teorileri Ve Türkiye Ekonomi- sinde 1990 Sonrasu Finansal Krizler. Marmara Üniversitesi İ.İ.B.F Dergisi, 1-22. Öksüz, S. (2001). Doğu Asya Mucizesinin Dünü Ve Kriz’in Bugünü: Türkiye İçin Bazı Çıkarımlar. Ege Akademik Bakış, 1(1), 39-64. Özdemir, İ. (2000). Asya Krizinin Turk Ve Dunya Ekonomisine Etkileri. İstanbul: Marmara Üniversitesi Sosyal Bilimler Ensititüsü İktisat Politikası Bilim Dalı. Schmukler, S., Kawai, M., & Newfarmer, R. (2001, Haziran). 342 \| Asya Krizi ve Krizi Aşmaya Yönelik Uygulanan Politikalar 1 Doktora Öğrencisi, Atatürk Üniversitesi, Sosyal Bilimler Enstitüsü, mehtabcelik2424@gmail.com, ORCID: http://orcid.org/0000-0002-8441-6278 2 Doç. Dr., Erzincan Binali Yıldırım Üniversitesi, İ.İ.B.F., Sağlık Yönetimi Bölümü, ekorkmaz@erzincan.edu.tr, ORCID: http://orcid.org/0000-0002-6787-5368 Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı ve Kârlılık Göstergeleri Açısından Analizi Mehtab Çelik1 Ersin Korkmaz2 Mehtab Çelik1 Ersin Korkmaz2 KAYNAKÇA Crisis And Con- tagion İn East Asia: Nine Lessons. Https://Elibrary.Worldbank.Org/: Htt- ps://Doi.Org/10.1596/1813-9450-2610 Adresinden Alındı Stiglitz, J. E. (2016). Küreselleşme: Büyük Hayal Kırıklığı. (A. Taşçıoğlu, & D. Vural, Çev.) İstanbul: Plan B. Sungur, N. (1999). Asya Krizinin Temel Dinamikleri. M.Ü. AT ENSTİTÜSÜ AVRUPA ARAŞTIRMALARI Dergisi, 203-212. Turgut, A. (2006). Finansal Krizlerde İMF’nin Rolü Ve Önemi 1997 Asya Ve 2000 - 2001 Türkiye Krizleri. Selçuk Üniversitesi Karaman İ.İ.B.F Dergisi, 1-14. Uygur, E. (2001, Nisan 7). Krizden Krize Türkiye: 2000 Kasım Ve 2001 Şubat Krizleri. Ankara, Çankaya, Türkiye. Bölüm 19 Özet Türkiye’nin jeopolitik konumu, coğrafi yapısı ve iklim şartlarının uygunluğu gibi koşullar sayesinde, Türkiye tarımsal ürünlerin üretiminde ve çeşitliliğinde önemli bir potansiyele sahiptir. Bu potansiyelin hayata geçirilerek üreticilerin sürdürülebilir ve verimli tarımsal üretim yapabilmeleri için temel faktör finansman kaynağıdır. Çiftçilerin finansman ihtiyaçlarını karşılayarak, ürünlerini değerlendirmek ve ihtiyaç duydukları üretim araç gereçlerin uygun maliyetle teminini sağlamak amacıyla, Türkiye Tarım Kredi Kooperatifleri kurulmuştur. Bu bağlamda, çalışma ile söz konusu kuruluşun finansal performansının likidite, finansal yapı ve kârlılık durumları açısından değerlendirilmesi amaçlanmıştır. Bu amaç doğrultusunda, kurumun 2018- 2022 dönemine ait faaliyet raporlarında yer alan konsolide bilanço ve gelir tablolarından elde edilen veriler oran analizi yöntemiyle analiz edilmiştir. Araştırma sonucunda, likidite göstergelerinden asit test oranının istenilen düzeyde, cari oranın düşük ve nakit oranının ise oldukça düşük seviyelerde olduğu tespit edilmiştir. Finansal yapı oranları değerlendirildiğinde, finansal kaldıraç oranının 2022 yılında oldukça yükseldiği dolayısıyla öz sermaye aktif oranının bu dönemde azaldığı, finansman oranının ise yıllar itibariyle genel kabul gören seviyenin oldukça altında gerçekleştiği belirlenmiştir. Son olarak kârlılık oranları incelendiğinde, brüt kâr marjının özellikle 2022 yılında oldukça azaldığı, net kâr marjının yalnızca 2021 yılında arttığı ve diğer yıllarda sürekli azaldığı, aktif kârlılık oranının ise 2019 yılı hariç diğer yıllarda artış gösterdiği tespit edilmiştir. https://doi.org/10.58830/ozgur.pub401.c1767 343 344 \| Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... 1. GİRİŞ İnsanoğlunun yaşamını devam ettirebilmesi için en önemli kaynaklardan biri gıda ve gıdanın üretildiği tarım sektörüdür. Temel ihtiyaçlardan olan gıda teminini doğal, sosyal ve ekonomik riskler altında üreten tarım sektörü, ülkelerin ekonomik ve sosyal gelişiminde önemli roller üstlenmektedir. Bu bağlamda ekonomik boyutunun ötesinde medeniyetlerin gelişiminde geçmişten günümüze stratejik öneme sahip bir sektör olarak değerlendirilebilir. Dolayısıyla tarımsal faaliyetlerin yürütülmesinin önünde engel teşkil edebilecek her türlü risklerin önceliklendirilerek yönetilmesi, tarımsal üretimde devamlılığın sağlanması ve üreticilerin karşılaştıkları finansman, tedarik ve pazarlama gibi sorunların çözüme kavuşturulması gerekir. Tarımsal faaliyetlerin toplumların refah düzeylerinin artmasındaki payına rağmen, günümüzde yaşanan küresel ısınma, aşırı nüfus artışı, tarımsal alanların atıl kalması, kuraklığın artması, doğal afetlerin yıkıcı etkisi ve orman yangınları gibi birçok sorun insanların temel yaşam ürünlerinin güvenliğini tehdit etmektedir. Ayrıca Covid-19 salgını, Rusya-Ukrayna savaşı ve ekonomik krizler gibi küresel ölçekte yaşanan olaylar, tüm ülkelerin tarım sektörünü öncelikli değerlendirilen sektörler arasına almalarını sağlamıştır. Türkiye’de de tarım alanında yaşanan bu gelişmeler çerçevesinde, tarımsal faaliyetlerin daha etkin ve verimli bir şekilde yürütülmesini sağlamak ve bu alanda dışa bağımlılığı azaltmak için Tarım ve Orman Bakanlığı, Ticaret Bakanlığı ve Tarım Kredi Kooperatifleri gibi kurumlar aracılığıyla birçok proje hayata geçirilerek desteklenmektedir. Türkiye’nin bulunduğu jeopolitik konum, coğrafi yapısı ve iklim şartlarının uygunluğu gibi koşullar sayesinde, Türkiye tarımsal ürünlerin üretiminde ve çeşitliliğinde önemli bir potansiyele sahiptir. Bu potansiyelin hayata geçirilerek üreticilerin sürdürülebilir ve verimli tarımsal üretim yapabilmeleri için temel faktör finansman kaynağıdır. Bu kapsamda cumhuriyet dönemi sonrası çiftçilerin finansman ihtiyaçlarını karşılayarak, ürünlerini değerlendirmek ve ihtiyaç duydukları üretim araç gereçlerin uygun maliyetle teminini sağlamak amacıyla Türkiye Tarım Kredi Kooperatifleri (TTKK)kurulmuştur. Kâr amacı taşımayan TTKK’lar özellikle küçük yerleşim yerlerinde yaşayan ve sermaye sıkıntısı bulunan çiftçilerin, orta ve küçük ölçekli tarım işletmelerin tarımsal faaliyetleri için ihtiyaç duydukları aynî ve nakdî finansman kaynaklarını düşük maliyetle ve uygun vadelerde karşılayarak, tarım sektörünün gelişmesine katkıda bulunmaktadırlar. TTKK’lar yardımlaşma, sorumluluk, demokrasi, eşitlik, hesap verme, adalet ve dayanışma ruhuylatarımsal faaliyetlerin henüz başlamadan önceki ve sonraki tüm süreçlerinde aktif ve öncü bir rol üstlenmektedir. Ayrıca TTKK’lar finansman sağlama faaliyetinin yanı sıra, kooperatif marketleri aracılığıyla tarımsal ürünlerin pazarının oluşturulması 345 Mehtab Çelik / Ersin Korkmaz \| ve tüketicilere daha uygun fiyatla ulaştırılmasını sağlamaktadır. Görüldüğü üzere bu örgütlenmeler, tarım sektöründe faaliyet gösteren birey veya işletmelere kaynak sağlama, üretilen ürünlerin ekiminden satışına kadar tüm süreçlerde önemli roller üstlenerek ülke ekonomisinin gelişmesine doğrudan katlı sunmaktadırlar. Bu kuruluşların gerekliliğini ve önemini farklı yönlerden ele alarak ortaya koymaya çalışan birçok araştırmada da elde edilen sonuçlar, yapılan bu değerlendirmeleri destekler niteliktedir. 1. GİRİŞ Bu denli önem arz eden kuruluşlar, her ne kadar kâr amacı taşımasalar da finansal gelişimlerinin değerlendirilmesi ve finansal açıdan güçlü ve zayıf yönlerinin ortaya konulması gerekir. Literatüre bakıldığında bölgesel veya il bazlı kooperatiflerin finansal performanslarının araştırmalara (Demir, 2003; Bücük, 2004; Seyfioğlu, 2010; Karaman ve diğ., 2014; Alıcı ve Demir, 2020) konu edinildiği görülmektedir. Ancak Türkiye Tarım Kredi Kooperatifleri’ni bu yönüyle inceleyen bir araştırmanın yapılmadığı söylenebilir. Bu bağlamda çalışma ile literatürdeki bu boşluğu gidermek ve TTKK kuruluşunun finansal gelişimini likidite, finansal yapı ve kârlılık durumları açısından değerlendirilmesi amaçlanmıştır. Bu amaç doğrultusunda ilk önce kooperatifçilik, TTKK’nın tarihçesi, organizasyonel yapısı ve faaliyet alanları teorik olarak ele alınmıştır. Daha sonra ise araştırmanın uygulama kısmında elde edilen bulgular değerlendirilerek ilgililere çeşitli önerilerde bulunulmuştur. 2. KOOPERATİFÇİLİK Latince kökenli “Cooperatio” kelimesinden gelen kooperatif kavramı, iş birliği ve uyum içinde çalışmayı ifade etmektedir (Mülayim, 2013: 57). Türk Dil Kurumu (TDK) kooperatifi; “üreticilerin aracıyı ortadan çıkararak ürünlerini daha iyi şartlarda pazarlamak için kurdukları ortaklık” şeklinde tanımlamaktadır (TDK, 2023). 1163 sayılı Kooperatifler Kanunu’nda kooperatif; “tüzel kişiliği haiz olmak üzere ortaklarının belirli ekonomik menfaatlerini ve özellikle meslek ve geçimlerine ait ihtiyaçlarını, işgücü ve parasal katkılarıyla karşılıklı yardım, dayanışma ve kefalet suretiyle sağlayıp korumak amacıyla gerçek ve tüzel kişiler tarafından kurulan değişir ortaklı ve değişir sermayeli ortaklıklar” olarak açıklanmaktadır (1163 sayılı Kooperatifler Kanunu). Bir başka tanımda ise kooperatifçilik, “birlikten kuvvet doğar” felsefesinden yola çıkarak, ülkelerin kalkınmasında ve bireylerin veya işletmelerin ekonomik sorunlarının çözülmesinde karşılıklı iş birliği, güç birliği ve yardımlaşma duygusuyla ekonomik imkânların bir araya getirilerek, ortak emek ve sermaye ile oluşturulan ticari girişimciliklerdir (Erkul vd., 2023:2). Toplumun sosyal ve ekonomik sorunlarını çözmek amacıyla yardımlaşma, eşitlik ve dayanışma ilkeleri çerçevesinde teşkilatlanan kooperatifler; tarım sektörünün ihtiyaç duyduğu üretim girdilerinin ve finansman kaynaklarının Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... 346 temininde, tarımsal üretim teknolojisinin geliştirilerek kullanılmasında ve bu sayede üretimde verimliliğin artırılmasında, satış işlemlerinin gerek üreticiler gerekse tüketiciler açısından en uygun şartlar altında gerçekleştirilmesinde etkin rol oynamaktadır. Ayrıca şehirlerde konut açığının giderilmesinde, esnaf ve sanatkârların tedarikten pazarlamaya kadar ekonomik anlamda daha uygun çalışma ortamlarının oluşturulmasında söz sahibi olmaları sebebiyle kooperatifler daha da önem kazanmaktadır (Yalçıntaş, 1970: 5). Kooperatiflerde temel amaç, özelikle sermaye yetersizliği sebebiyle bireylerin tek başlarına üstesinden gelemeyecekleri işleri en az maliyetle, yardımlaşma ve dayanışma duygusuyla yapmaktır. Bireylerin benzer konular veya fikirler etrafında toplanmaları sonucunda Esnaf-Kefalet Kooperatifleri, Tarımla İlgili Kooperatifler, Yapı Kooperatifleri gibi farklı amaçlarla kooperatiflerin kurulması gerçekleşir (Güngör, 2023: 25). Kooperatifler hangi amaç doğrultusunda kurulursa kurulsun hepsinin ortak noktası, üyelerinin “belirli” ekonomik menfaatlerini, meslek ve geçimleri ile ilgili ihtiyaçlarını karşılamaktır. Buradaki “belirli” ifadesi kooperatiflerin ana sözleşmesi ile belirlenmiş olan amaç ve konu hükümleri için önem arz etmektedir. Bu bağlamda, kooperatifler amaç ve hedefleri bakımından kâr amacı taşıyan ideal gayeler benimsemiş derneklerden ve kâr etme amacı dışında kurulmuş olması ile de ticari şirketlerden farklılaşmaktadır. Bu yönüyle kooperatiflerde üyeler arasında iş birliği ve bunun neticesinde de toplumsal dayanışma ön plana çıkmaktadır (Geray, 1991: 12). 2. KOOPERATİFÇİLİK Genel olarak kooperatiflerin amaçlarına ve ön plana çıktığı konulara bakıldığında, üyeler arasındaki fikir uyuşmazlıklarını aşarak sağlıklı bir düzenin oluşturulmasının temelini ortaya koyan kooperatifçiliğin hedefleri; sosyal açıdan gelişmeye katkıda bulunmak, sosyal adaletin yaygınlaşmasını sağlamak, sosyal barışa destek olmak, sosyal dengeyi korumak, sosyal bütünleşmeye katkıda bulunmak ve demokrasinin yaygınlaşmasına imkân sağlamak şeklinde sıralanmaktadır (Güven, 1997: 26). Toplumsal kalkınmayı geliştirerek ülke ekonomisine katkıda bulunan kooperatifler, genel olarak aşağıdaki özellikleri bünyesinde barındırırlar (Civan 2021: 42): • Kooperatifler, ortakları yardımlaşma ve dayanışma içerisinde olmak için bir araya getirmektedir. • Kooperatifler tüzel kişiliğe sahiptir. • Gerçek ve tüzel kişiler tarafından kurulabilirler. • Kooperatiflerin ortak sayısında sınırlama yoktur. • Kooperatiflerde ortaklar arasında eşitlik prensibi vardır. • Kooperatiflerde sermaye payına bakılmaksızın her ortak eşit oy hakkına sahiptir. • Kooperatiflerde sermaye payına bakılmaksızın her ortak eşit oy hakkına sahiptir. Mehtab Çelik / Ersin Korkmaz \| 347 Mehtab Çelik / Ersin Korkmaz \| 347 347 Genel anlamda kuruluş amacı Türk çiftçisine hizmet etmek olan TTKK’ların çalışma konuları Tarım Kredi Kooperatifi Ana Sözleşmesi’nde (Madde 5) aşağıdaki gibi sıralanmıştır (TTKK, 2023a): 9 “Ortakların: 9 “Ortakların: 9 “Ortakların: • Kısa ve orta vadeli kredi ile her türlü tarımsal girdi ihtiyaçlarını karşılamak, • Mahsullerini değerlendirmek ve pazarlamak, • Mahsullerini değerlendirmek ve pazarlamak, • Müştereken faydalanabilecekleri her çeşit makine, ekipman ve tesisleri temin etmek. 9 Ortak ve gerektiğinde diğer üreticilerin: 9 Ortak ve gerektiğinde diğer üreticilerin: • Üretim ve zarurî tüketim maddeleri ile üretim araçlarını toptan sağlamak ve bu maksatla, Merkez Birliği yönetim kurulunun müsaadesi ile kuruluşlara iştirak etmek, • Hükümetçe görev verildiği takdirde, Devletçe yapılacak destekleme ödemelerine aracılık etmek, • El sanatlarını geliştirmek ve mamullerini değerlendirmek, • Meslekî ve teknik yönden bilgilerini arttırmak, sosyal ve kültürel konularla ilgili faaliyetlerde bulunmak, kurslar açmak ve seminerler tertip etmek. 9 Mevduat toplamak, bankacılık hizmetleri ve sigorta acenteliği yapmak. 9 1581 Sayılı Kanun ile ek ve değişikliklerine göre kurulmuş diğer kooperatifler, Bölge Birlikleri ve Merkez Birliği ile kredi ihtiyaçlarını karşılamak üzere banka ve sigorta şirketi kurmak veya bu gibi kuruluşlara iştirak etmek, 9 Bölge birliğince kendisine verilecek diğer görevleri yapmak. 9 Bölge birliğince kendisine verilecek diğer görevleri yapmak. Kooperatif bu faaliyetlerini, üst kuruluşlarca tespit edilecek genel uygulama esaslarına göre yürütür.” 3. TÜRKİYE TARIM KREDİ KOOPERATİFLERİNİN İ İ Tasarruf Menafi Sandıkları ile devam eden Türk Kooperatifçilik Hareketi, Cumhuriyetin ilanından sonraki dönemde 1924’te, 498 sayılı İtibari Zirai Birlikler Kanunu, 1929’da 1470 sayılı Zirai Kredi Kooperatifleri Kanunu ile gelişimini devam ettirmiş ve 1935 yılında çıkarılan 2836 sayılı Tarım Kredi Kooperatifleri Kanunu ile TTKK’nın kuruluşu gerçekleştirilmiştir (İnan, 2008: 36).1972 yılında çıkarılan 1581 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu ile bölge birlikleri ve merkez birliğinin kurulmasına zemin oluşturulmuştur. Böylece TTKK’nın dikey organizasyonel yapısının alt yapısı oluşturulmuştur. TTKK, daha önce T.C. Ziraat Bankası çatısı altında faaliyetlerini sürdürürken, 1977 yılında merkez birliğinin kurulmasıyla birlikte tam bağımsız bir çiftçi kuruluşu sıfatını kazanmıştır. 1984 yılına gelindiğinde, 237 sayılı Kanun Hükmünde Kararname (KHK) ile 1581 sayılı kanunda değişiklik yapılarak, TTKK’ların Yönetim Kurulları kendi ortakları içerisinden seçilerek oluşturulmaya başlanmıştır. Bu düzenleme demokratik kooperatifçilik alanında önemli bir adım olarak görülmektedir. Takip eden yılda 237 sayılı KHK değiştirilerek, 3223 sayılı 1581 Sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanununun bazı maddelerini değiştiren 237 Sayılı Kanun Hükmünde Kararnamenin değiştirilerek kabulü hakkında kanun hazırlanmıştır. Bu kanun kapsamında TTKK, Tarım ve Orman Bakanlığının bir kuruluşu olmuş ve denetimleri de yine bu bakanlığın sorumluluğuna verilmiştir. Yapılan düzenlenmeler uygulanmaya başlandıkça merkez birliğinde genel müdürün, bölge birliklerinde ise bölge müdürünün yönetim kurulu başkanlıklarının olması nedeniyle çeşitli yönetim sıkıntıları ortaya çıkmıştır. Bunun sonucunda, 1995 yılında 553 sayılı KHK ile TTKK’ların idari yapısında önemli değişiklikler yapılarak 1581 sayılı kanun ile öngörülen düzenlemelere tekrar geri dönülmüştür. Daha sonra 2005 tarihinde yürürlüğe giren 5330 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu ile kurumun ihtiyaç duyduğu yasal alt yapının son hali verilmiştir. Bu düzenlemeler sayesinde kooperatifler daha demokratik hale gelmiştir (TTKK, 2023b). 3. TÜRKİYE TARIM KREDİ KOOPERATİFLERİNİN İ İ TTKK’nın tarihçesi,1863 yılında dönemin Niş valisi (Sırbistan’ın güneyinde yer alan bir şehir) Mithat Paşa tarafından Şehir Köyü kasabasında Türk geleneğinde bulunan karşılıklı yardımlaşma ve dayanışma prensipleri örnek alınarak kurulan Memleket Sandıkları ile başlamaktadır. 1867’de Memleket Sandıkları Nizamnâmesi’nin kullanımı ile Osmanlı Devleti’nin dört bir yanında kurulan bu sandıklar yıllar boyu başarılı bir şekilde faaliyetlerini sürdürmüşlerdir (Güngör, 2023: 25).Cumhuriyetten önceki dönemde Memleket Sandıkları ile başlayan ve daha sonra 1883yılında kurulan 348 \| Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... Tasarruf Menafi Sandıkları ile devam eden Türk Kooperatifçilik Hareketi, Cumhuriyetin ilanından sonraki dönemde 1924’te, 498 sayılı İtibari Zirai Birlikler Kanunu, 1929’da 1470 sayılı Zirai Kredi Kooperatifleri Kanunu ile gelişimini devam ettirmiş ve 1935 yılında çıkarılan 2836 sayılı Tarım Kredi Kooperatifleri Kanunu ile TTKK’nın kuruluşu gerçekleştirilmiştir (İnan, 2008: 36).1972 yılında çıkarılan 1581 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu ile bölge birlikleri ve merkez birliğinin kurulmasına zemin oluşturulmuştur. Böylece TTKK’nın dikey organizasyonel yapısının alt yapısı oluşturulmuştur. TTKK, daha önce T.C. Ziraat Bankası çatısı altında faaliyetlerini sürdürürken, 1977 yılında merkez birliğinin kurulmasıyla birlikte tam bağımsız bir çiftçi kuruluşu sıfatını kazanmıştır. 1984 yılına gelindiğinde, 237 sayılı Kanun Hükmünde Kararname (KHK) ile 1581 sayılı kanunda değişiklik yapılarak, TTKK’ların Yönetim Kurulları kendi ortakları içerisinden seçilerek oluşturulmaya başlanmıştır. Bu düzenleme demokratik kooperatifçilik alanında önemli bir adım olarak görülmektedir. Takip eden yılda 237 sayılı KHK değiştirilerek, 3223 sayılı 1581 Sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanununun bazı maddelerini değiştiren 237 Sayılı Kanun Hükmünde Kararnamenin değiştirilerek kabulü hakkında kanun hazırlanmıştır. Bu kanun kapsamında TTKK, Tarım ve Orman Bakanlığının bir kuruluşu olmuş ve denetimleri de yine bu bakanlığın sorumluluğuna verilmiştir. Yapılan düzenlenmeler uygulanmaya başlandıkça merkez birliğinde genel müdürün, bölge birliklerinde ise bölge müdürünün yönetim kurulu başkanlıklarının olması nedeniyle çeşitli yönetim sıkıntıları ortaya çıkmıştır. Bunun sonucunda, 1995 yılında 553 sayılı KHK ile TTKK’ların idari yapısında önemli değişiklikler yapılarak 1581 sayılı kanun ile öngörülen düzenlemelere tekrar geri dönülmüştür. Daha sonra 2005 tarihinde yürürlüğe giren 5330 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu ile kurumun ihtiyaç duyduğu yasal alt yapının son hali verilmiştir. Bu düzenlemeler sayesinde kooperatifler daha demokratik hale gelmiştir (TTKK, 2023b). 4. TÜRKİYE TARIM KREDİ KOOPERATİFLERİNİN ORGANİZASYON YAPISI 800.000 ortağı ile 1 Merkez Birliği, 17 bölge birliği, 1618 kooperatif, 192 hizmet bürosu ve 18 iştiraki ile faaliyetlerine geniş bir çalışma alanında devam eden TTKK’nın organizasyon yapısı aşağıda genel hatlarıyla açıklanmıştır (TTKK, 2023c). Birim Tarım Kredi Kooperatifleri: Çiftçileri finansal açıdan destekleyerek mevcut durumlarını iyileştirmek ve ihtiyaç duydukları girdileri temin etmek amacıyla kurulan birim tarım kredi kooperatiflerinin en az 30 349 Mehtab Çelik / Ersin Korkmaz \| Mehtab Çelik / Ersin Korkmaz ortağı bulunmalıdır. Bu kooperatiflerin kurulmasına üst birlik olan merkez birliği karar vermekte ve kurulan kooperatifler faaliyet gösterdikleri alan içerisinde yer alan bölge birliğine bağlı faaliyetlerini sürdürmek zorundadırlar. 1581 sayılı kanunun 21. maddesine göre tescil ve ilâna tâbi olmayan bu kooperatiflerin organizasyon şemasında en yetkili organ olan genel kurul, 5 üyeden oluşan ve 4 yıllığına seçilen yönetim kurulu, genel kurul temsilcileri arasından 4 yıllığına seçilen ve 2 denetçiden oluşan denetçiler organı yer almaktadır (TTKK, 2023a). Bölge Birlikleri: En az 30 kooperatifin bir araya gelmesi ile kurulabilen bölge birliklerinin amacı, birliğe ortak olan kooperatiflerin menfaatlerini korumak ve kooperatifçilik faaliyetlerinin gelişmesine katkıda bulunmaktır. Bir bölge birliğinin kurulmasına Tarım ve Orman Bakanlığından izin alındıktan sonra merkez birliğince karar verilir. Bölge birliklerinin organizasyon şemasında en yetkili organ olan genel kurul, 7 üyeden oluşan ve 4 yıllığına seçilen yönetim kurulu, genel kurul temsilcileri veya kooperatif ortakları arasından 4 yıllığına seçilen ve 2 denetçiden oluşan denetçiler organı yer almaktadır (TTKK, 2023d). Merkez Birliği: Bu birlik, tarım kredi kooperatifleri ile bölge birliklerinin ortak menfaatlerini korumak, amaçlarını gerçekleştirmeleri için faaliyette bulunmak ve her türlü çalışmalarını düzenlemek üzere 1581 sayılı kanun ile görevlendirilmiştir. Merkez birliği kurulabilmesi için 5 bölge birliğinin ortaklığı şartı söz konusudur. Merkez birliğinin organizasyon yapısında en yetkili organ olan ve bağlı bölge birliklerinin kendi genel kurullarında seçilen temsilciler ile merkez birliği yönetim kurulu üyelerinden oluşan genel kurul, genel kurul üyeleri arasından seçilen 9 üyeden oluşan ve 4 yıllığına seçilen yönetim kurulu, genel kurulda farklı bölge birlikleri temsilcileri arasından seçilen 2 denetçi ve Tarım ve Orman Bakanlığı tarafından atanan 1 denetçiden oluşan ve 4 yıllığına atanan denetçiler organı yer almaktadır (TKKK, 2023e) İştirakler: Merkez birliği, kooperatif ve bölge birliklerinin çıkarlarını korumak, tohumlar, gübreler, ilaçlar ve çeşitli ekipmanlar gibi üretim ve zaruri tüketim maddelerini tedarik etmek, ürünlerin değerlendirilmesi ile ilgili faaliyetleri rasyonel bir şekilde düzenlemek ve bu konularda her türlü teşebbüse girişmek amacıyla şirketler tesis edebilir veya bu tür şirketlere iştirak olabilir (1581 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu). 4. TÜRKİYE TARIM KREDİ KOOPERATİFLERİNİN ORGANİZASYON YAPISI 2022 yılı sonu itibariyle TTKK’nın 18 iştiraki ve hissedarı olduğu 23 şirket bulunmaktadır. Bu şirketlerden 19’unun sermayesinin %50 ve daha fazlası TTKK’ya ait olup bağlı ortaklığı konumundadır (TTKK, 2023f). 350 Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... 5. TARIM KREDİ KOOPERATİFLERİNİN FAALİYETLERİ Muhasebe Faaliyetleri: TTKK’nın tek düzen muhasebe sistemine geçişi 1998 yılında; merkez birliği, bölge birliği ve birim kooperatiflerin bilgisayarlı muhasebe sistemine geçişi ise 2000 yılında olmuştur. Muhasebe verilerinde tek düzeliği, doğruluğu, güvenilirliği ve denetlenebilirliği sağlamak amacıyla 2009 yılında tüm birimlerin muhasebe işlemlerinin birbirine entegreli ve tek bir merkezden yürütülmesini sağlayan bir ERP yazılımı olan e-koop bilgi sistemi kullanılmaya başlanmıştır (Akıncı, 2019: 12). Kredi Faaliyetleri: Birim tarım kredi kooperatifleri üyelerinin kısa ve orta vadeli her türlü ihtiyaçlarını gidermek, devletçe sağlanan sübvansiyonlu tarımsal kredileri çiftçilere ulaştırmak ve tarımsal üretimi geliştirmek için üyelerine kredi kullandırmaktadır. Bu kapsamda 2022 yılı faaliyet raporuna göre 2019-2022 yılları arasında kullandırılan kredi bilgileri Tablo 1’de verilmiştir (TTKK, 2022: 17). Tablo 1: Kredi Dağılımı ve Kredi Kullanan Ortak Sayısı Yıllar 2019 2020 2021 2022 Kullandırılan Kredi 8.381.070.504 8.260.967.862 11.036.293.101 26.084.910.479 İndirimli Krediler 5.706.147.312 6.272.611.495 9.195.503.431 22.903.426.735 Ortak Sayısı 390.763 364.238 356.683 396.274 Tablo 1: Kredi Dağılımı ve Kredi Kullanan Ortak Sayısı Sigortacılık Faaliyetleri: TTKK ana sözleşmesine göre teşkilatın ana faaliyetlerinden bir tanesi de sigorta faaliyetidir. Türkiye Sigorta Birliği (TSB) tarafından yayımlanan verilerine göre; 2022 yılında 23 şirketin bulunduğu Tarsim Bitkisel Ürün Sigortası sektöründe toplam 4.010.553.341 TL üretim yapılmıştır. Bu üretimin 1.140.875.197 TL’lik kısmı TTKK aracılığıyla gerçekleştirilmiştir. Prim tutarı bazında TTKK’nın sektör payı %28 olarak gerçekleşmiştir. 2022 yılında Türkiye genelinde Tarsim Bitkisel Ürün Sigortası sektöründe 2.249.573 adet poliçe üretilmiş bunun 1.085.462 adedi TTKK tarafından gerçekleştirilmiştir (TTKK, 2022: 18). Pazarlama Faaliyetleri: Bölge birlikleri ve bağlı birim kooperatifler üreticilerin, gübre, bitki besleme, zirai mücadele ilacı, halk sağlığı ürünleri, yem, tohumluk, fide-fidan, akaryakıt, traktör, büyükbaş ve küçükbaş ile diğer hayvanların temin edilmesi noktasında finansman sağlarken, ilgili ürünler ortaklara kooperatif tarafından teslim edilerek, bu ürünlerin bedelleri firmalara ya da anlaşmalı kurumlara ortaklar adına ödenmektedir. 2022 yılında TTKK 351 Mehtab Çelik / Ersin Korkmaz teşkilatı tarafından 52.258.695.663 TL satış gerçekleştirilmiştir. Bu satışlarda en büyük payı kimyevi gübre, motorin ve karma yem kalemleri almaktadır. Ayrıca TTKK’lar tarafından 2022 yılı sonu itibariyle 12.314.943.594 TL tutarında 1.607.554 ton ürün alımı yapılmıştır (TTKK, 2022: 19). Denetim Faaliyetleri: Merkez birliği Tarım ve Orman Bakanlığının (TTKK, 2023e), bölge birlikleri hem merkez birliğinin hem de Tarım ve Orman Bakanlığının (TTKK, 2023a), kooperatifler ise bölge birliği, merkez birliği ve Tarım ve Orman Bakanlığının teftiş ve denetimine tabidirler. Yapılan teftiş ve denetimler sonucunda kanun ve ana sözleşmelerdeki görevleri ile ilgili ortaya çıkan eksiklikler konusunda denetlenen kurum denetleyen kurumun vereceği talimatlara uymak zorundadır (TTKK, 2023d). Bu kurumlarda yapılan denetimler daha çok uygunluk denetimi şeklinde icra edilmektedir. 5. TARIM KREDİ KOOPERATİFLERİNİN FAALİYETLERİ TTKK’larda kurumsal risk yönetimi ve iç denetim faaliyetleriyle ilgili uygulamalar henüz başlangıç seviyesinde olmakla beraber, kooperatiflerde risk yönetimi ve risk odaklı denetim kültürünün yerleşmesi için çeşitli çalışmalar yapılmaktadır (Uysal, 2020: 702). Ayrıca kooperatiflerde kullanılan e-koop otomosyon sistemi sayesinde muhasebe, krediler, stok, insan kaynakları gibi tüm iş süreçlerinde hata yapma oranı azalmakta ve bu sayede merkezi denetim sistemi kapsamında işlemlerin takip ve kontrolü uzaktan gerçekleştirilerek ortaya çıkabilecek hata veya hileli işlemlerin tespiti ve önlenmesi sağlanmaktadır (Kayacık, 2023: 19). 6. ARAŞTIRMANIN KAPSAMI VE YÖNTEMİ 6. ARAŞTIRMANIN KAPSAMI VE YÖNTEMİ Araştırmanın kapsamını TTKK kurumunun web sitesinde yayımlanan 2018-2022 dönemine ait faaliyet raporlarında yer alan enflasyon düzeltmesine tâbi tutulmuş konsolide bilanço ve gelir tabloları oluşturmaktadır. Söz konusu finansal tablolardan elde edilen verilerin analizinde literatürde yaygın olarak kullanılan ve finansal tablolarda incelenmek istenen kalemler arasındaki basit oransal ilişkileri görmek ve yorumlamak amacıyla kullanılan oran analizi yöntemi kullanılmıştır (Akgüç, 2011: 463). Bu kapsamda TTKK’ların finansal performansının değerlendirilmesi için literatürde bu amaçla yaygın olarak kullanılan likidite, finansal yapı ve kârlılık oranları kullanılmıştır. Likidite oranları, kurumun kısa vadeli borçlarını ödeme gücünü ve beklenmedik piyasa koşullarında faaliyetlerini devam ettirebilme yetisini gösteren oranlardır (Ekinci ve Bakır, 2021: 6). Araştırma kapsamında kullanılan likidite oranlarının açıklamaları aşağıda verilmiştir (Elmas, 2017: 205-207): • Cari Oran: Bu oran işletmenin dönen varlıkları ile kısa vadeli borçları arasındaki ilişkisini gösterir. Bir başka ifadeyle işletmenin kısa vadeli • Cari Oran: Bu oran işletmenin dönen varlıkları ile kısa vadeli borçları arasındaki ilişkisini gösterir. Bir başka ifadeyle işletmenin kısa vadeli 352 Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... borçlarını ödeme gücünü ifade eder. Cari oranın genel olarak 2 olması yeterli görülürken, Türkiye gibi gelişmekte olan ülkelerde 1,5’a kadar düşmesi normal kabul edilebilir. borçlarını ödeme gücünü ifade eder. Cari oranın genel olarak 2 olması yeterli görülürken, Türkiye gibi gelişmekte olan ülkelerde 1,5’a kadar düşmesi normal kabul edilebilir. borçlarını ödeme gücünü ifade eder. Cari oranın genel olarak 2 olması yeterli görülürken, Türkiye gibi gelişmekte olan ülkelerde 1,5’a kadar düşmesi normal kabul edilebilir. • Asit- Test Oranı: Bu oran işletmenin dönen varlık kalemlerinden nakde dönüşüm hızı en düşük olan stoklar kalemi dışındaki dönen varlıkların kısa vadeli borcunu ödeme gücünü ifade eder. Bu oranın genel olarak 1 olması yeterli görülürken, Türkiye gibi gelişmekte olan ülkelerde 0,70’e kadar düşmesi normal kabul edilebilir. • Nakit Oran: Bu oran işletmenin stoklarını elden çıkaramaması veya alacaklarını tahsil edememesi durumunda mevcut nakit ve nakit benzeri varlıklar ve finansal yatırımlarla kısa vadeli borçlarının ne kadarını ödeyebileceğini ifade eder. Nakit oran, likide oranları içinde en hassas olan orandır. Bu oranın genel olarak 0,20’nin altına düşmemesi arzu edilir ancak Türkiye gibi gelişmekte olan ülkelerde 0,20’nin altına düşebilmektedir. Finansal yapı oranları, işletmelerin kaynak yapısını ve uzun vadeli borç ödeme gücünü temsil eden göstergelerdir. İşletmenin kaynaklarının aktiflerin fonlamasında nasıl kullanıldığı, borç öz sermaye dağılımı, kaynak kullanımının optimalliği, finansal yapının sağlamlığı ve katma değer oluşturmadaki başarıları bu oranlar ile ölçülmektedir (Ekinci ve Bakır, 2021: 13). 6. ARAŞTIRMANIN KAPSAMI VE YÖNTEMİ Aynı zamanda finansal yapı oranları, işletmelerin faaliyetleri sonucunda zarar etmesi, varlıkların değerinde düşüş yaşanması veya gelecek yıllarda tahmin edilen nakit akımının gerçekleşmemesi durumunda yükümlülüklerin yerine getirilip getirilemeyeceği hakkında önemli ipuçları verir (Akgüç, 2011: 447). Çalışmada kullanılan finansal yapı oranlarının açıklamaları aşağıda verilmiştir (Akgüç, 2011: 450-452). • Finansal Kaldıraç Oranı: Bu oran, varlıkların yüzde kaçının yabancı kaynaklar yoluyla finanse edildiğini ifade eder. Oranın yüksek olması, işletmenin riskli bir şekilde finanse edildiğini, faiz ve borçların ödenememesi nedeniyle mali açıdan zor durumda olma olasılığının yüksek olduğunu, kredi verenlerce emniyet marjının dar olduğunu gösterir. • Öz Kaynak Aktif Oranı: Bu oran işletmenin varlıklarının yüzde kaçının işletme sahip veya ortakları tarafından finanse edildiğini ifade eder. İşletmenin mali gücünü gösteren bu oranın eğilimi bir işletmenin uzun vadeli borç ödeme kapasitesini ortaya koymaktadır. İşletmelerde bu oranın yüksek olması işletmenin faiz yükünün genel olarak düşük olduğu ve uzun vadeli borçlarını ödeyememe ihtimalinin 353 Mehtab Çelik / Ersin Korkmaz az olduğu şeklinde yorumlanır. Bir başka ifadeyle işletmenin genel olarak finansal açıdan iyi bir şekilde yönetildiğinin göstergesi olarak değerlendirilebilir. • Finansman Oranı: İşletmenin finansal açıdan bağımsızlığını gösteren bu oran, aynı zamanda işletmenin öz kaynakları ile toplam borçları arasında uygun bir ilişkinin olup olmadığını gösterir. Bu oranın en az 1 olması beklenir. Kredi veren kurum veya kuruluşlar açısından finansman oranının büyük olması arzu edilirken, işletme yöneticileri ise bu oranın düşük olmasını arzu etmektedirler. Varlıkların, likiditenin ve borçların ne kadar etkin yönetildiğinin bir göstergesi olan kârlılık oranları, idarî politika ve kararların sonuçlarını göstermesi nedeniyle işletmelerin toplam finansal performansının ölçüsü niteliğini taşımaktadır (Gapenski, 2005: 532). Bu oranlar, işletmenin kâr elde etmedeki genel verimliliğini ölçmektedir (Weaver, 2012: 62). Araştırma kapsamında kullanılan kârlılık oranlarının açıklamaları aşağıda verilmiştir (Savcı, 2011: 222; Akdoğan ve Tenker, 2010: 669). • Brüt Kâr Marjı Oranı: Satışlara göre kârlılığın belirlenmesinde kullanılan bu oran, işletmenin net satışlarının yüzde kaçının brüt satış kârı olduğunu göstermesi açısından önemlidir. İşletmelere üretim maliyetlerinin kontrolü ve fiyatlama politikası için bilgi sağlamak amacıyla hesaplanan brüt kâr marjı oranının yüksek olması veya yükselme trendi göstermesi, olumlu bir durum olarak değerlendirilmektedir. • Net Kâr Marjı Oranı: Bu oran, satışların kârlılık düzeyini veya her 1 TL’lik net satıştan ne kadar kâr elde edildiğini göstermektedir. Oranın yüksek olması işletme açısından olumlu bir gösterge olarak yorumlanır. • Aktif Kârlılık Oranı Bu oran, varlıkların işletmede ne ölçüde etkin kullanıldığını veya varlıların kâr oluşturabilme kapasitesini görmek bakımından önem taşımaktadır. Genellikle işlemenin aktif kârlılığının yüksek olması beklenir. 6. ARAŞTIRMANIN KAPSAMI VE YÖNTEMİ Araştırma kapsamında kullanılan likidite, finansal yapı ve kârlılık oranlarının hesaplanması ile ilgili formüller Tablo 2’de sunulmuştur. 354 \| Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... Tablo 2: Likidite, Finansal Yapı ve Kârlılık Oranları Likidite Oranları Formüller Cari Oran Dönen Varlık / Kısa Vadeli Yabancı Kaynak Asit-Test Oranı Dönen Varlık- Stoklar / Kısa Vadeli Yabancı Kaynak Nakit Oranı Hazır Değerler / Kısa Vadeli Yabancı Kaynak Finansal Yapı Oranları Finansal Kaldıraç Oranı Toplam Yabancı Kaynak / Aktif Toplam Öz Kaynak Aktif Oranı Öz Kaynaklar / Aktif Toplam Finansman Oranı Öz Kaynaklar /Toplam Yabancı Kaynaklar Kârlılık Oranları Brüt Kar Marjı Brüt Satış Kârı / Net Satışlar Net Kar Marjı Net Kâr / Net Satışlar Aktif Karlılık Oranı Net Kâr / Aktif Toplam Tablo 2: Likidite, Finansal Yapı ve Kârlılık Oranları 7. BULGULAR Araştırmanın bu kısmında TTKK’nın likidite, finansal yapı ve kârlılık oranlarına ilişkin elde edilen bulguların analiz ve değerlendirmelerine yer verilecektir. 7.2. Finansal Yapı Oranları Açısından Analiz Sonuçları TTKK’nın finansal yapı oranlarına ilişkin hesaplanan finansal kaldıraç oranı, öz kaynak aktif oranı ve finansman oranı Tablo 4’te yer almaktadır. Tablo 4: Finansal Yapı Oranları Finansal Yapı Oranları 2018 2019 2020 2021 2022 Finansal Kaldıraç Oranı 0,74 0,70 0,67 0,74 0,81 Öz Kaynak Aktif Oranı 0,26 0,30 0,33 0,26 0,19 Finansman Oranı 0,36 0,44 0,49 0,35 0,24 Tablo 4’e bakıldığında, TTKK’nın finansal kaldıraç oranının 2022 yılı hariç diğer yıllarda Türkiye için kabul edilen oranın 0,60’ın da üzerinde olduğu göz önünde bulundurulduğunda normal olarak kabul edilen seviyededir. Ancak 2022 yılında kurumun finansal kaldıraç oranının oldukça yükseldiği ve 0,81 olarak hesaplandığı görülmektedir. Öz kaynak aktif oranına bakıldığında ise, kurumun bu oranı ele alınan dönem itibari ile Türkiye koşullarına göre anılan 0,40 oranının altında seyretmektedir. Bu oran özellikle 2022 yılında 0,19 ile en düşük seviyeye ulaşmıştır. Kurumun yıllar itibari ile hesaplanan finansman oranları incelendiğinde Türkiye gibi gelişmekte olan ülkeler için normal kabul edilen 0,70 oranının da oldukça altında kaldığı görülmektedir. Bu oran 2020 yılına kadar artarak devam etmiş, 2022 yılında ise 0,24 ile en düşük seviyesini görmüştür. 7.1. Likidite Oranları Açısından Analiz Sonuçları TTKK’nın likidite oranlarına ilişkin hesaplanan cari oran, asit-test oranı ve nakit oranı Tablo 3’te sunulmuştur. Tablo 3: Likidite Oranları Likidite Oranları 2018 2019 2020 2021 2022 Cari Oran 1,21 1,27 1,33 1,25 1,18 Asit-Test Oranı 1,16 1,21 1,27 1,11 1,08 Nakit Oranı 0,003 0,007 0,012 0,011 0,012 Tablo 3’e bakıldığında, çalışmada ele alınan dönem boyunca TTKK’nın cari oranının Türkiye için genel olarak kabul edilen 1,5 oranının altında olduğu görülmektedir. Aynı zamanda cari oranın 2020 yılına kadar artış gösterdiği ancak daha sonraki yıllarda düşüş trendine girdiği belirlenmiştir. 355 Mehtab Çelik / Ersin Korkmaz Kurumun beşinci faaliyet yılında cari oranı 1,18 olarak hesaplanmıştır. Bu sonuç, kurumun her 1 TL’lik kısa vadeli borcuna karşılık 1,18 TL’sinin olduğunu göstermektedir. Cari oranın tamamlayıcısı niteliğinde olan asit- test oranının ise, söz konusu dönem boyunca genel olarak arzu edilen seviye olan 1’in üzerinde seyrettiği görülmektedir. Bu sonuç, stoklar dışındaki dönen varlıkların kısa vadeli borçları ödemek için yeterli olduğunu göstermektedir. Ayrıca asit test oranının da 2020 yılına kadar artış gösterdiği ancak daha sonraki yıllarda düşüş eğilimine girdiği belirlenmiştir. Kurumun nakit oranının seyrine bakıldığında ele alınan dönem boyunca genel kabul gören oran olan 0,20’den oldukça düşük olduğu görülmektedir. Bu durum, kurumun ciddi bir nakit problemi olduğu sonucunu göstermektedir. Kurumun beşinci faaliyet yılında cari oranı 1,18 olarak hesaplanmıştır. Bu sonuç, kurumun her 1 TL’lik kısa vadeli borcuna karşılık 1,18 TL’sinin olduğunu göstermektedir. Cari oranın tamamlayıcısı niteliğinde olan asit- test oranının ise, söz konusu dönem boyunca genel olarak arzu edilen seviye olan 1’in üzerinde seyrettiği görülmektedir. Bu sonuç, stoklar dışındaki dönen varlıkların kısa vadeli borçları ödemek için yeterli olduğunu göstermektedir. Ayrıca asit test oranının da 2020 yılına kadar artış gösterdiği ancak daha sonraki yıllarda düşüş eğilimine girdiği belirlenmiştir. Kurumun nakit oranının seyrine bakıldığında ele alınan dönem boyunca genel kabul gören oran olan 0,20’den oldukça düşük olduğu görülmektedir. Bu durum, kurumun ciddi bir nakit problemi olduğu sonucunu göstermektedir. 7.3. Kârlılık Oranları açısından Analiz Sonuçları TTKK’nın kârlılık oranlarına ilişkin hesaplanan brüt kâr marjı, net kâr marjı ve aktif kârlılık oranı Tablo 5’te verilmiştir. \| Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... 356 Tablo 5: Kârlılık Oranları Kârlılık Oranları 2018 2019 2020 2021 2022 Brüt Kâr Marjı 8,84 8,83 9,26 8,50 5,67 Net Kâr Marjı 5,21 4,47 4,62 6,12 5,35 Aktif Kârlılık Oranı 3,31 3,01 3,09 4,49 5,45 Tablo 5: Kârlılık Oranları Tablo 5 incelendiğinde, TTKK’nın brüt kâr marjının en yüksek 2020 yılında gerçekleştiği, daha sonra azalma eğilimine girerek en büyük düşüşün 2022 yılında gerçekleştiği görülmektedir. Kurumun net kâr marjının ise 2019 ve 2022 yıllarında azaldığı, 2021 yılında en yüksek seviyesini gördüğü belirlenmiştir. Aktif kârlılık oranının yalnızca 2019 yılında azaldığı, diğer yıllarda büyümeye devam ettiği tespit edilmiştir. 8. SONUÇ 19. yüzyıldan itibaren modern bir işletme olarak kabul edilen kooperatifler hem ortaya çıkış nedenleri hem de çalışma şekillerindeki farklılıklar ve benimsemiş olduğu ilkeler nedeniyle yalnızca bir şirket türü olarak değerlendirilmemektedir. Sadece ekonomik değil, aynı zamanda kültürel ve sosyal alanlarda da hizmet sağlayan kooperatifler, ticaret kanunlarında da yer edinerek önemli iktisadî aktörler haline gelmişlerdir. Küresel ekonomik krizlerin yaşandığı dönemlerde dahî faaliyetlerini sürdürmeyi başaran kooperatifler; yaygınlaşan yoksulluk, gittikçe artan istihdam talebi, kültürel ve sosyal yozlaşma gibi sorunlarla etkin bir şekilde mücadele edilmesini sağladığı için ülkelerin gündemindeki yerini her geçen gün daha da artırmaktadır. Diğer yandan kooperatifler; âtıl kaynakların ekonomiye kazandırılmasına, sermayenin tabana yayılmasına, nispeten düşük gelirli kesimin kendini güvenceye alarak iktisadî faaliyetlerde bulunmasına, küçük girişimlerin bir araya gelerek büyük aktörlere dönüşmesine, ortakların sürdürülebilir bir ekonomik gelire sahip olmalarına dolayısıyla ekonomik istikrarın sağlanmasına katkıda bulunmaktadırlar. Sosyal, kültürel ve ekonomik faaliyetlerde bulunarak toplumun çeşitli ihtiyaçlarına cevap veren kooperatifler, kamu ile özel sektör arasında üçüncü bir sektör olarak kabul görmektedir. Bireylerin ferdî olarak karşılamakta güçlük çekecekleri ihtiyaçlarını karşılıklı iş birliği, dayanışma ve yardımlaşma yoluyla gidermek amacıyla kurulan kooperatifler toplumsal bir nitelik taşırlar. Bu anlamda insan odaklı olarak kurulan kooperatiflerin amacı; en az maliyetle, en avantajlı şekilde, faaliyette bulunulan bölgenin sosyal, kültürel ve ekonomik şartları da göz önünde tutularak sürdürülebilir stratejilerle ortaklarının 357 Mehtab Çelik / Ersin Korkmaz \| Mehtab Çelik / Ersin Korkmaz ihtiyaçlarına cevap vermek olarak belirlenmiştir. Çok yönlü yapıya sahip olan bu kooperatiflerin sürdürülebilirliğinin sağlanması noktasında finansal performanslarının dikkate alınması önem taşımaktadır. Bu doğrultuda çalışma ile, Türkiye Tarım Kredi Kooperatiflerinin likidite, finansal yapı ve kârlılık durumları analiz edilerek değerlendirmelerde bulunulmuştur. Çalışma sonucunda, kurumun kısa vadeli borçlarını ödeyebilme gücünü gösteren rasyolardan yalnızca asit test oranının ele alınan dönem boyunca istenilen seviyede olduğu, cari oranın gelişmekte olan ülkeler için kabul gören seviyenin de altında olduğu ve özellikle en hassas likidite ölçüsü olan nakit oranının oldukça düşük seyrettiği görülmektedir. Kurumun işletme sermayesinin, genel olarak yüksek kısa vadeli yabancı kaynakla finanse edilmesi nedeniyle cari oranın düşük olduğu söylenebilir. Nakit oranının da oldukça düşük olduğu göz önünde bulundurulduğunda, kurumun alacaklarının tahsilinde sorun yaşaması ya da satışlarının durması halinde kısa vadeli borçlarını ödeme gücünün çok düşük olduğu ifade edilebilir. Özellikle 2020 yılından sonra kurumun, daha fazla likidite sorunu ile karşı karşıya kaldığı belirtilebilir. Bu bağlamda likiditeyi artırmaya yönelik girişimlerde bulunulmalıdır. Kurumun finansal yapı oranları değerlendirildiğinde, 2022 yılında toplam varlıkların yüksek ölçüde yabancı kaynaklarla finanse edildiği, dolayısıyla kurumun fonlama riskinin arttığı söylenebilir. 8. SONUÇ Bu durumla ilişkili olarak da kurumun toplam varlıklar içerisindeki öz sermaye payının düşük olduğu belirlenmiştir. Dolayısıyla kurumun alacaklarının tahsilinde yaşayacağı bir güçlükte, fiyatlarda görülen beklenmedik düşüşlerde ya da ekonominin genelinde görülebilecek bir durgunluk döneminde finansal açıdan zor durumda kalabileceği söylenebilir. Kurumun finansman oranı ise yıllar itibariyle genel kabul gören oranın oldukça altındadır. Bu oranlar kurumun ciddi bir kaynak sıkıntısı yaşadığını ortaya koyar niteliktedir. Kurumun kârlılık oranları incelendiğinde, brüt kâr marjının özellikle 2022 yılında oldukça azaldığı görülmektedir. Bu dönemde yüksek enflasyonun maliyetleri artırması nedeniyle bu sonuçla karşılaşılmış olabilir. Kurumun net kâr marjı ise yalnızca 2021 yılında artmış, diğer yıllarda sürekli düşüş eğilimi göstermiştir. Büyük ölçüde yabancı kaynak kullanan bu kurumların kaynak maliyetinin yüksek olması ve buna bağlı olarak net kârının azalması bu sonucun ortaya çıkmasında neden olabilir. Aktif kârlılık oranına bakıldığında 2019 yılı hariç diğer yıllarda bu oranın artış gösterdiği tespit edilmiştir. Elde edilen bu sonuçlar, genel olarak kurumun satışlara göre kârlılığının azaldığını ancak varlıklarının daha etkin ve verimli bir şekilde kullanılarak varlık getirisinin artırıldığını göstermektedir. 58 \| Türkiye Tarım Kredi Kooperatiflerinin Finansal Performansının Likidite, Finansal Yapı... 358 KAYNAKÇA 1163 sayılı Kooperatifler Kanunu, https://www.mevzuat.gov.tr/mevzuatme- tin/1.5.1163.pdf, Erişim Tarihi: 19.11.2023 1581 sayılı Tarım Kredi Kooperatifleri ve Birlikleri Kanunu, https://www.mev- zuat.gov.tr/mevzuat?MevzuatNo=1581&MevzuatTur=1&MevzuatTer- tip=5, Erişim Tarihi: 27.11.2023 Akdoğan, N. ve Tenker, N. (2010). Finansal Tablolar ve Mali Analiz Teknikleri. Ankara: Gazi Kitapevi. Akgüç, Ö. (2011). Mali Tablolar Analizi. (14. Baskı) İstanbul: Arayış Basım ve Yayıncılık. Akıncı, S. (2019). Tarım Kredi Kooperatiflerinde Merkezi Muhasebe Sistemi Uygulamasının İncelenmesi. Dumlupınar Üniversitesi İİBF Dergisi, (3-4), 8-18. Alıcı, M.S. & Demir, M. (2020). 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Tarım Kredi Kooperatiflerinin Mali Yapısının Tespiti ve So- runlarının Çözümüne Yönelik Öneriler. (Yüksek Lisans Tezi). Gazi Üni- versitesi/Sosyal Bilimler Enstitüsü, Ankara. Ticaret Bakanlığı, (2023). Türkiye’de Kooperatifçilik, https://ticaret.gov.tr/ kooperatifcilik/kooperatifler-hakkinda-bilgiler/tarihce, Erişim tarihi: 21.11.2023. Türk Dil Kurumu, (2023). Türk Dil Kurumu Sözlükleri, https://sozluk.gov.tr/, Erişim Tarihi: 21.11.2023. Türkiye Tarım Kredi Kooperatifleri, (2023e). Tarım Kredi Kooperatifleri Merkez Birliği Ana Sözleşmesi, https://www.tarimkredi.org.tr/media/ zl5j45cc/turkiye-tarim-kredi-kooperatifleri-merkez-birligi-ana-sozlesme- si.pdf, Erişim Tarihi: 27.11.2023. Türkiye Tarım Kredi Kooperatifleri, (2022). 2022 Faaliyet Raporu, https:// www.tarimkredi.org.tr/media/a0gfbz0q/faaliyet_raporu_2023.pdf, Eri- şim Tarihi: 28.11.2023. Türkiye Tarım Kredi Kooperatifleri, (2023a). 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Türkiye Tarım Kredi Kooperatifleri İçin Kurumsal Risk Yönetimi ve İç Denetim Model Önerisi. Uluslararası Yö- netim Akademisi Dergisi, 3(3), 698-708. Weaver, S. C. (2012). The Essentials of Financial Analysis, Mcgraw Hill Com- panies, USA. Yalçıntaş, N. (1970). Kooperatiflerle İlgili Olarak Devlete Düşen Görevler. VII. Türk Kooperatifçilik Kongresi. Ankara: Türk Kooperatifçilik Kurumu. Bölüm 20 2 Doktora Öğrencisi, Selçuk Üniversitesi Sosyal Bilimler Enstitüsü, aysegulgudul@hotmail.com, ORCID ID: 0000-0002-7217-1947 1 Doktora Öğrencisi, Selçuk Üniversitesi Sosyal Bilimler Enstitüsü, asliaydin1710@outlook.com, ORCID ID: 0000-0002-9354-2591. Özet Günümüz dünyasında teknolojinin hızla ilerlemesi ve pazardaki gelişmeler, geleneksel postacılık hizmetlerinin yeniden yapılandırılmasını zorunlu kılmıştır. Özellikle haberleşme ağının dijitalleşmesiyle, kişiler daha erişilebilir ve ulaşılabilir uygulamalara yönelmektedir. Bu süreç posta, haberleşme ve kargo işletmelerinin web sitelerindeki kullanıcı dostu arayüzlerden, takip sistemlerine, müşteri desteğinden ödeme seçeneklerine kadar geniş bir yelpazede değerlendirilmesine olanak sağlamakla birlikte daha etkili çevrimiçi hizmet sunmasına yönelik fırsat yaratacaktır. Çalışmada, Türkiye’de faaliyet gösteren ve en fazla tercih edilen kurumsal posta, haberleşme ve kargo işletmelerine ait web sitelerin içerik zenginliğini ölçmek için TOPSIS yöntemi kullanılmıştır. Bu yöntem çok kriterli karar verme problemlerinin çözümünde etkilidir. Bu yöntemle işletmelerin web sitelerin sıralanması için kullanım kolaylığı, kurumsal bilgi, sayfa içeriği ve yönlendirme olmak üzere 4 ana kriter belirlenmiştir. Bu kriterlerden kullanım kolaylığı, web sitesinin ziyaretçilerinin rahatlıkla gezinip istedikleri bilgilere ulaşabilmelerini; kurumsal bilgi kriteri, işletme hakkında sunulan bilgilerin doğruluğu ve kapsamlılığın; sayfa içeriği, web sitesinin içerdiği bilgilerin çeşitliliği ve kapsamlılığın; yönlendirme kriteri ise web sitesinin kullanıcıları başka sayfalara veya içeriklere yönlendirebilme becerisini ölçmektedir. Analize en çok tercih edilen 5 posta, haberleşme ve kargo işletmesi dahil edilmiş olup, işletmelerin web siteleri incelenmiş ve belirlenen kriterler kapsamında içerik zenginliklerine göre sıralanmıştır. https://doi.org/10.58830/ozgur.pub401.c1768 361 362 Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 1. Posta, Haberleşme ve Kargo İşletmeleri Geçmiş dönemlerde haberleşme ihtiyacı, kamu otoritesi ve posta tekeli yapısıyla sunulan hizmetlerle karşılanmaktaydı. Bu dönemde yapılan araştırmalar genellikle posta hizmetlerinin maliyet yapısıyla ilgili olmakla birlikte, aynı zamanda bu endüstrinin doğal bir tekel olup olmadığı sorusu üzerinde duruluyordu. Eğer doğal bir tekelse, rekabetin faydalı olmayacağı düşünülüyordu. Ancak zaman içinde bazı araştırmacılar, posta hizmetinin belirli alt segmentlerinde rekabetin başlatılmasının refah kazanımlarına yol açabileceğini savunmaya başlamış, hatta posta hizmetinin bütün olarak doğal bir tekel olmasına rağmen bu alt segmentlerde rekabetin faydalı olabileceğini de öne sürmüşlerdi (Rogerson ve Takis, 1993:109). Örneğin PTT, önceleri sadece posta hizmeti verirken, günümüzde birçok hizmet vermektedir. Özellikle taşımacılık ve bankacılık hizmetleri bakımından günümüzde hizmet veren PTT’de taşımacılık; posta, kargo ve lojistik hizmetlerinden oluşmakla birlikte, posta ve kargo hizmetleri, rekabete açılmış ve “son adım teslimat” anlayışı ile yürütülmektedir. Bankacılık hizmetleriyse mevduat toplamanın dışında, birçok bankacılık işlemlerinin yapıldığı temel finansal hizmetlerini içermektedir (PTT, 2023). Gelişen ve değişen teknolojik koşullar göz önünde bulundurulduğunda, haberleşme amacıyla kullanılan posta hizmetlerinin de geliştiği ve değiştiği görülmektedir. Günümüz postacılık hizmetlerinin azalması hem kişi hem de kurumları kargo ve lojistik hizmeti veren işletmelere yönlendirmektedir. Türk Dil Kurumu’nun Büyük Türkçe Sözlüğü’ne (2023) göre kargo “Bir yerden bir yere yük veya posta taşıyan şirket” ve “Bir şirketin taşıdığı yük veya posta” şeklinde ifade edilmiştir. Bu ifadeden hareketle kargo taşımacılığı, bir yük veya postanın gönderildiği noktadan alınıp hedef noktaya ulaştırılmasını içeren faaliyetler bütünüdür (Ulutaş ve Karaköy, 2019: 224). Bu faaliyetler, göndericiden alınan yükün, depolanmasından ve uygun bir şekilde paketlenmesinden sonra belirlenen adres veya noktaya ulaştırılmasını içermektedir. Büyük Türkçe Sözlüğe (2023) göre lojistik ise “Kişilerin ihtiyaçlarını karşılamak üzere her türlü ürünün, hizmetin ve bilgi akışının çıkış noktasından varış noktasına kadar taşınmasının etkili ve verimli bir biçimde planlanması ve uygulanması” şeklinde ifade edilmiştir. Kamunun ötesinde posta ve haberleşme hizmetleri günümüzde neredeyse tamamen özel sektör ağırlıklı yapılmaktadır. Özellikle haberleşmenin geniş bir ağ içerisinde gerçekleştirilmeye başlanmasıyla, sunulan hizmetin tüm bölgelere erişimi genişletilmiş, posta ve haberleşme hizmetleri kargo ve lojistik hizmetleri ağırlık olarak yeniden yapılandırılan küresel bir yapı haline gelmiştir. Bununla birlikte, yapılanan bu yeni sistem yatırım için Aslı Uğur Aydın Taştekin / Ayşegül Güdül 363 cazip bir pazar haline gelmiştir (Bilgi Teknolojileri ve İletişim Kurumu- Sektörel Araştırma ve Strateji Geliştirme Dairesi Başkanlığı, 2011: 11-15). Yenilenen bu yapının teknolojik kolaylıklarla desteklenmesi, bu işletmelerin çeşitliliğini artırmakla birlikte, daha erişilebilir kıldığından müşteri talebine daha hızlı yanıt verebilmektedir. 1. Posta, Haberleşme ve Kargo İşletmeleri Özellikle bu işletmelerin erişilebilir ve ulaşılabilir olmalarını kolaylaştırma adına, kurumsal web sitelerinin sayfa içeriği, kurumsal bilgi ve yönlendirme gibi unsurların her kitleye uygun, anlaşılabilir, sade ve açıklayıcı olması müşterilerine kullanım kolaylığı sağlamaktadır. 2. Metodoloji 2.1. Çalışmanın Amacı, Önemi ve Katkıları Posta, haberleşme ve kargo işletmelerinin web siteleri, kullanıcı deneyimi, bilgi erişilebilirliği, sayfa içeriği ve gezinme kolaylığı gibi kritik unsurları içerir. Dolayısıyla sektörde web sitelerin kullanıcı dostu arayüzleri, izlenebilirlik ve takip sistemleri, gönderi takibi, müşteri hizmetleri ve ödeme seçenekleri gibi operasyonel özellikler değerli addedilmektedir. Bu dikkate alınarak ve TOPSIS yöntemi kullanılarak yapılan bu çalışmanın temel amacı, posta, haberleşme ve kargo işletmelerinin web sitelerini kapsamlı bir şekilde analiz ederek, bu sitelerin kullanım kolaylığı, kurumsal bilgi, sayfa içeriği ve yönlendirme kriterlerine dair performanslarını değerlendirmektir. Çalışmanın önemi, posta, haberleşme ve kargo işletmelerinin dijital varlıklarının günümüzün dijital pazarlama ortamında ne kadar kritik olduğunu anlamak ve bu işletmelerin web sitelerinin, müşteri memnuniyeti, rekabet avantajı ve kurumsal itibar gibi faktörler üzerindeki etkisini belirginleştirecektir. Bu açıdan ele alındığında çalışma, sektördeki web sitelerinin güçlü yönlerini ve geliştirilmesi gereken alanları belirleyerek, sektör işletmelerinin daha etkili çevrimiçi hizmet sunmalarına yardımcı olacaktır. Bu, aynı zamanda sektördeki rekabeti anlamak ve müşteri beklentilerine daha iyi yanıt verebilmek için önemli bir yol haritası sunacaktır. Ayrıca, bu çalışma, sektördeki diğer işletmelerin web sitelerini iyileştirmek için bir referans noktası olarak hizmet edebileceği düşünülmektedir. Çalışmanın katkıları, posta, haberleşme ve kargo işletmelerinin dijital pazarlama stratejilerini geliştirmelerine ve web sitelerinin kullanıcı deneyimini artırmalarına yardımcı olabilecek en iyi uygulamaları belirlemeye yöneliktir. Bu çalışma, sektördeki web sitelerinin mevcut durumunu değerlendirerek, sektördeki en iyi uygulamaları ve iyileştirme potansiyellerini ortaya koyarak sektöre önemli bir katkı sağlayabilir. 364 4 \| Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 2.2. Çalışmanın Evren ve Örneklemi Çalışmanın evreni, Türkiye’de faaliyet gösteren posta, haberleşme ve kargo işletmeleridir. Örneklem ise Türkiye’de faaliyet gösteren kurumsal işletmelerden en çok bilinirliğe sahip ve web sitesi açısından daha kullanışlı olan 5 işletmeden oluşmaktadır. 2.3.Çalışmanın Yöntemi Çalışmada belirlenen kriterlere göre posta, haberleşme ve kargo sektöründe faaliyet göstermekte olan işletmelerin web site içerik zenginliğini sıralamak için TOPSIS yönteminden (İdeal Çözüme Benzerlik Bakımından Sıralama Tekniği) yararlanılmıştır (Pavic ve Novoselac, 2013). TOPSIS yöntemi, pozitif ideal çözüme en yakın alternatifi en iyi alternatif olarak kabul etmektedir (Doğanalp, 2012; Sağır ve Doğanalp, 2016; Sağır ve Güdül, 2023). Bu yöntemde amaç, birden fazla kritere sahip olan seçenekler arasından en ideal olanı belirlemektir. Yöntem altı adımdan oluşmakta ve bu adımların sırasıyla uygulanması sonucunda seçenekler ideallik sıralamasına göre sıralanmaktadır (Pavic ve Novoselac, 2013). Posta, haberleşme ve kargo sektöründe TOPSIS yönteminin birçok alanda kullanıldığı görülmektedir. Bunlara sosyal medya platformu aracılığı ile müşteri yorumları ve şikayetlerinin ele alındığı çalışmalar (Burucuoğlu ve Erdoğan-Yazar, 2020; Atılgan ve Yoğurtcu, 2021; Hirata ve Matsuda, 2023) ve çok kriterli karar verme yöntemlerinin kullanıldığı benzer çalışmalardan (Öztürk vd., 2008; Kannan vd., 2009; Bhatti vd., 2010; Uzun, 2013; Ding vd., 2014; Önden vd., 2015; Asoğlu ve Eren, 2018; Ulutaş ve Karaköy, 2019; Lazarevic vd., 2020; Putri ve Sari, 2021; Şenaran, 2023) örnek gösterilebilir. Çalışma örneklemini oluşturan posta, kargo ve haberleşme işletmelerinin web sitelerini analiz etmek için kriterler kullanım kolaylığı, kurumsal bilgi, sayfa içeriği ve yönlendirme olarak belirlenmiş ve bunlar kriterler için alt kriterler belirlenmiştir. Kullanım kolaylığı, web sitesinin ziyaretçilerinin rahatlıkla gezinip istedikleri bilgilere ulaşabilmelerini; kurumsal bilgi kriteri, işletme hakkında sunulan bilgilerin doğruluğu ve kapsamlılığın; sayfa içeriği, web sitesinin içerdiği bilgilerin çeşitliliği ve kapsamlılığın; yönlendirme kriteri ise web sitesinin kullanıcıları başka sayfalara veya içeriklere yönlendirebilme becerisini ölçmektedir. Kullanım kolaylığı alt kriterleri kullanım kolaylığı, kendine ait URL ismi, farklı dil seçenekleri, kullanışlı menü, ana sayfaya kolay erişim, arama butonu, siteye üyelik, kullanım koşulları ve güvenlik-gizlilik bilgisidir. Kurumsal bilgi alt kriterleri hakkımızda, işletmenin tarihçesi, güncel haber ve yenilikler, vizyon ve misyon, kariyer olanakları/İK, kalite çalışmaları, ödüller, 365 Aslı Uğur Aydın Taştekin / Ayşegül Güdül yatırımcı ilişkileri, işletme logosu ve son güncelleme tarihidir. Sayfa içeriği alt kriterleri şube isimleri, şubelere erişim ya da bağlantı, ulaşım bilgisi, haritada gösterim, fiyat bilgisi, evden kargo alımı, online ödeme, güncel promosyon duyuruları, e-posta bülteni, çevrimiçi ziyaretçi defteri, çevrimiçi iletişim/ online destek, geri bildirim formu, web sayfa değerlendirme seçeneği ve yardım menüsüdür. Yönlendirme kriterlerinin alt kriterleri ise sosyal medya bağlantıları, şubelere ilişkin bilgi ve tanıtım ve sıkça sorulan sorulardır. TOPSIS yönteminin adımları şu şekildedir: Adım 1: Seçenek ve kriterlerin bulunduğu karar matrisinin oluşturulması. D= Adım 2: Karar matrisinin oluşturulmasından sonra, normalleştirilmiş karar matrisinin oluşturulması. 2.3.Çalışmanın Yöntemi Adım 3: Adım 2’de oluşturulan normalleştirilmiş karar matrisinde yer alan değerlerin belirlenmiş olan kriter ağırlıklarıyla çarpılması sonucunda elde edilen ağırlıklı normalleştirilmiş karar matrisinin oluşturulması. Adım 4: Ağırlıklı normalleştirilmiş karar matrisine göre aşağıdaki formüllerle pozitif ideal (en iyi) ve negatif ideal (en kötü) çözümlerin belirlenmesi. 366 \| Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 366 Adım 5: Pozitif ve negatif çözümlere uzaklığın bulunması. Adım 5: Pozitif ve negatif çözümlere uzaklığın bulunması. Adım 6: İdeal çözüme göreli yakınlıkların hesaplanması ve sıralamanın oluşturulması. Adım 6: İdeal çözüme göreli yakınlıkların hesaplanması ve sıralamanın oluşturulması. Analize dahil edilen 5 posta, haberleşme ve kargo işletmeleri İ1, İ2, İ3, İ4 ve İ5 olarak isimlendirilmiştir. Çalışmaya dahil edilen bu işletmeler hala hizmet vermeye devam ettikleri için isimleri açıkça belirtilmemiştir. 2.4. Bulgular 2.4. Bulgular TOPSIS yönteminde kullanılmak için gerekli olan kriter ağırlıkları Tablo 1’de gösterilmektedir. Tablo 1: Kriter Ağırlıkları Kullanım Kolaylığı Kurumsal Bilgi Sayfa İçeriği Yönlendirme 0,38986 0,06792 0,38986 0,15235 Belirlenen alt kriterlerden posta, haberleşme ve kargo işletmelerinin kaç tanesine sahip olduğunun belirlenmesi için yapılan web site incelemeleri çerçevesinde oluşturulan karar matrisi Tablo 2’de gösterilmektedir. Tablo 2: Karar Matrisi Kriterler Kullanım Kolaylığı Kurumsal Bilgi Sayfa İçeriği Yönlendirme İ1 6 3 7 3 İ2 6 6 6 3 İ3 8 8 9 4 İ4 5 5 7 3 İ5 6 5 7 2 Aslı Uğur Aydın Taştekin / Ayşegül Güdül \| 367 Karar matrisinin oluşturulmasından sonra, Adım 2’de gösterilen formülün uygulanmasıyla, normalleştirilmiş karar matrisi oluşturulmuştur. Normalleştirilmiş karar matrisi Tablo 3’te gösterilmektedir. Karar matrisinin oluşturulmasından sonra, Adım 2’de gösterilen formülün uygulanmasıyla, normalleştirilmiş karar matrisi oluşturulmuştur. Normalleştirilmiş karar matrisi Tablo 3’te gösterilmektedir. Tablo 3: Normalleştirilmiş Karar Matrisi Kriterler Kullanım Kolaylığı Kurumsal Bilgi Sayfa İçeriği Yönlendirme İ1 0,427482 0,237915 0,43082 0,437595 İ2 0,427482 0,475831 0,369274 0,437595 İ3 0,569976 0,634441 0,553912 0,58346 İ4 0,356235 0,396526 0,43082 0,437595 İ5 0,427482 0,396526 0,43082 0,29173 Tablo 3: Normalleştirilmiş Karar Matrisi Normalleştirilmiş karar matrisinde elde edilen verilerin belirlenen kriter ağırlıklarıyla çarpılması sonucunda elde edilen ağırlıklı normalleştirilmiş karar matrisi Tablo 4’te gösterilmektedir. Tablo 4: Ağırlıklı Normalleştirilmiş Karar Matrisi Tablo 4: Ağırlıklı Normalleştirilmiş Karar Matrisi Kriterler Kullanım Kolaylığı Kurumsal Bilgi Sayfa İçeriği Yönlendirme İ1 0,166658 0,016159 0,16796 0,066668 İ2 0,166658 0,032318 0,143965 0,066668 İ3 0,222211 0,043091 0,215948 0,08889 İ4 0,138882 0,026932 0,16796 0,066668 İ5 0,166658 0,026932 0,16796 0,044445 Adım 4’te gösterilen formülün uygulanmasıyla elde edilen pozitif ve negatif ideal çözümler Tablo 5’te gösterilmektedir. Tablo 5: Pozitif ve Negatif İdeal Çözümler Kullanım Kolaylığı Kurumsal Bilgi Sayfa İçeriği Yönlendirme Pozitif (A+) 0,222211 0,043091 0,215948 0,08889 Negatif (A-) 0,138882 0,016159 0,143965 0,044445 Adım 5’te gösterilen formülün uygulanmasıyla pozitif ve negatif ideal çözümlere uzaklığın bulunmasıyla belirlenen ayrım ölçüleri Tablo 6’da gösterilmektedir. \| Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 368 Tablo 6: Ayrım Ölçüleri S+ S- İ1 0,081291 0,042908 İ2 0,094221 0,03907 İ3 0 0,121762 İ4 0,100008 0,034433 İ5 0,087324 0,038253 Seçeneklerin ideal çözüme göreli yakınlıkları Tablo 7’de gösterilmiştir. Tablo 6: Ayrım Ölçüleri S+ S- İ1 0,081291 0,042908 İ2 0,094221 0,03907 İ3 0 0,121762 İ4 0,100008 0,034433 İ5 0,087324 0,038253 Seçeneklerin ideal çözüme göreli yakınlıkları Tablo 7’de gösterilmiştir. Seçeneklerin ideal çözüme göreli yakınlıkları Tablo 7’de gösterilmiştir. Seçeneklerin ideal çözüme göreli yakınlıkları Tablo 7’de gösterilmiştir. 2.4. Bulgular Tablo 7: İdeal Çözüme Göreli Yakınlık Tablo 7: İdeal Çözüme Göreli Yakınlık İşletme Sonuç İ1 0,345478 İ2 0,293121 İ3 1 İ4 0,525089 İ5 0,304619 İ İdeal çözüme göre yakınlıkları hesaplanan posta, haberleşme ve kargo işletmelerinin sıralaması Tablo 8’de gösterilmektedir. Tablo 8: İşletme Sıralaması Sıra İşletme Sonuç 1 İ3 1 2 İ4 0,525089 3 İ1 0,345478 4 İ5 0,304619 5 İ2 0,293121 Yapılan analiz sonucunda 1. sırada İ3, 2. sırada İ4, 3. sırada İ1, 4. sırada İ5 ve 5. sırada ise İ2’nin olduğu görülmektedir. SONUÇ VE ÖNERİLER Çalışmada Türkiye’de faaliyet göstermekte olan posta, haberleşme ve kargo işletmelerinin web sitelerinin içerik zenginliğinin değerlendirilmesi amaçlanmıştır. Belirlenen kriterler üzerinden yapılan TOPSIS sonucunda web siteleri idealliğine göre sıralanmıştır. Çeşitli faktörler göz önüne alındığında 369 Aslı Uğur Aydın Taştekin / Ayşegül Güdül çalışmanın, sektördeki rekabet avantajı, verimlilik ve müşteri memnuniyeti açısından önemli olduğu düşünülmektedir. Çalışmada, belirlenen kriterleri en çok karşılayan işletmenin belirlendiği gibi posta, haberleşme ve kargo işletmelerinin web sitelerindeki güçlü ve zayıf yönlerini anlamasına katkı sağlayacağı düşünülmektedir. TOPSIS yöntemiyle web site içerik zenginlikleri analiz edilen 5 posta, haberleşme ve kargo işletmeleri arasından en ideal olarak seçilen İ3 olmuştur. Karar matrisi incelendiğinde bu işletmenin kullanım kolaylığı alt kriterlerinden 6, kurumsal bilgi alt kriterlerinden 3, sayfa içeriği alt kriterlerinden 7 ve yönlendirme alt kriterlerinde 3 tanesinin web sitesinde yer aldığı görülmektedir. Genel olarak kriterler incelendiğinde diğer posta, haberleşme ve kargo işletmelerinden daha fazla kriterin web sitesinde bulunduğu görülmektedir. Diğer posta, haberleşme ve kargo işletmelerinden farklı olarak web sitesinde sunduğu hizmetler ise farklı dil seçenekleri ve kullanım koşulları olarak sıralanmaktadır. İkinci sırada yer alan İ4’e ait bilgiler karar matrisinde incelendiğinde, kullanım kolaylığı alt kriterlerden 5, kurumsal bilgi alt kriterlerinden 5, sayfa içeriği alt kriterlerinden 7, yönlendirme alt kriterlerinden 3 tanesinin web sitesinde yer aldığı görülmektedir. Birinci sırada yer alan İ3 ve İ4’ün karar matrisi karşılaştırıldığında, tüm kriterleri İ3’e göre daha az karşıladığı görülmektedir. Üçüncü sırada yer alan İ1’e ait bilgiler karar matrisinde incelendiğinde, kullanım kolaylığı alt kriterlerden 5, kurumsal bilgi alt kriterlerinden 3, sayfa içeriği alt kriterlerinden 7, yönlendirme alt kriterlerinden 3 tanesinin web sitesinde yer aldığı görülmektedir. Kullanım kolaylığı kriterinde İ4’e göre daha fazla alt kriteri sağlasa da diğer kriterleri daha az sağladığı görülmektedir. 1. sırada yer alan İ3 ile karşılaştırıldığında ise sayfa içeriği kriterinde aynı sayıda kriteri sağlasa da diğer kriterleri daha az sağladığı görülmektedir. Dördüncü sırada yer alan İ5’e ait bilgiler karar matrisinde incelediğinde, kullanım kolaylığı alt kriterlerden 6, kurumsal bilgi alt kriterlerinden 5, sayfa içeriği alt kriterlerinden 7, yönlendirme alt kriterlerinden 2 tanesinin web sitesinde yer aldığı görülmektedir. Dördüncü sırada yer alan İ1 ile karşılaştırıldığında, yönlendirme alt kriterlerinde daha azının web sitesine bulundurmasından dolayı ve yönlendirme kriterinin kriter ağırlığının daha yüksek olmasından dolayı sıralama bu şekilde olmuştur. Son olarak beşinci sırada yer alan İ2’ye ait bilgiler karar matrisinde incelendiğinde, kullanım kolaylığı alt kriterlerden 6, kurumsal bilgi alt kriterlerinden 6, sayfa içeriği alt kriterlerinden 6, yönlendirme alt Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... SONUÇ VE ÖNERİLER 372 \| Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 372 SONUÇ VE ÖNERİLER 370 kriterlerinden 3 tanesinin web sitesinde yer aldığı görülmektedir. Kurumsal bilgi kriterlerinin ikinci, üçüncü ve dördüncü sırada yer alan işletmelere göre daha fazla bulunmasına rağmen bu kriterin kriter ağırlığının daha düşük olmasından dolayı, bu işletme son sırada yer almıştır. Posta, haberleşme ve kargo işletmeleri genel olarak incelendiğinde hepsinde yer alan özelliklerin kendine ait URL ismi, ana sayfaya kolay erişim, kullanışlı menü, güvenlik/gizlilik bilgisi, güncel haber ve yenilikler, işletme logosu, fiyat bilgisi ve sosyal medya bağlantıları olduğu görülmektedir. İşletmelerin çoğunda olmayan kriterler ise işletme tarihçesi, ödüller, kalite çalışmaları, yatırımcı ilişkileri, ulaşım bilgisi ve haritada gösterim, çevrimiçi ziyaretçi defteri ve geri bildirim formudur. Elde edilen sonuçlar doğrultusunda sektör ve akademik çalışmalar için aşağıdaki öneriler sunulabilir: 1. İçerik Zenginliği Geliştirme: En iyi performans gösteren İ3 işletmesinin web sitesi incelenerek, diğer işletmelerin içeriklerindeki eksiklikler ve güçlü yönler belirlenebilir. Özellikle eksik olan alanlarda içerik geliştirme çalışmaları yapılabilir. Örneğin, işletmeler tarihçelerini, ödüllerini veya kalite çalışmalarını daha detaylı bir şekilde paylaşarak içeriklerini zenginleştirebilirler. 2. Kullanıcı Dostu Arayüz Tasarımı: İşletmeler, kullanıcı deneyimini iyileştirmek için web sitelerinin navigasyonunu kolaylaştırabilir. Kullanıcıların aradıkları bilgilere daha hızlı erişebilmeleri için daha basit ve anlaşılır bir arayüz tasarlayabilirler. 3. Çeşitlilik ve Dil Seçenekleri: İ3 işletmesinin farklı dil seçeneklerini sunması dikkate değer bir durum olduğundan diğer işletmeler de uluslararası kullanıcı kitlesini hedefleyerek web sitelerinde farklı dil seçeneklerini veya kültürel çeşitliliği yansıtan unsurları ekleyebilirler. 3. Çeşitlilik ve Dil Seçenekleri: İ3 işletmesinin farklı dil seçeneklerini sunması dikkate değer bir durum olduğundan diğer işletmeler de uluslararası kullanıcı kitlesini hedefleyerek web sitelerinde farklı dil seçeneklerini veya kültürel çeşitliliği yansıtan unsurları ekleyebilirler. 4. Güncel Tutma ve İletişim Kanalları: İşletmeler, güncel haberleri, yenilikleri düzenli olarak paylaşarak ziyaretçilerin ilgisini canlı tutabilir. Ayrıca, sosyal medya bağlantıları ve geri bildirim formu gibi iletişim kanallarıyla kullanıcılarla etkileşimi artırabilirler. 5. Tüm İşletmelerin Analize Dâhil Edilmesi: Gelecekte yapılacak çalışmalarda daha kapsamlı bir örneklem kullanarak Türkiye’deki tüm posta, haberleşme ve kargo işletmelerinin analize dâhil edilmesi, daha geniş bir perspektif sunabilir ve sektördeki genel eğilimleri gösterme açısından daha değerli olabilir. 371 Aslı Uğur Aydın Taştekin / Ayşegül Güdül \| Çalışmanın örneklemi Türkiye genelinde en çok tercih edilen kurumsal 5 posta, haberleşme ve kargo işletmesinden oluştuğu için sınırlılıkları bulunmaktadır. İlerleyen zamanlarda yapılacak çalışmalarda Türkiye’de faaliyet gösteren tüm posta, haberleşme ve kargo işletmelerinin analize dahil edilmesi, ana kriter ve alt kriterlerin arttırılması veya uluslararası faaliyet gösteren posta, haberleşme ve kargo işletmelerinin analize dahil edilmesinin hem işletmelerin web sitelerini geliştirmesine hem de tercih edilebilirliklerini arttırmasına olanak sağlayacağı düşünülmektedir. KAYNAKÇA Asoğlu, İ., & Eren, T. (2018). AHP, TOPSIS, PROMETHEE Yöntemleri ile Bir İşletme İçin Kargo Şirketi Seçimi. Yalova Sosyal Bilimler Dergisi, 8(16), 102-122. Atılgan, K. Ö., & Yoğurtcu, H. (2021). Kargo Firması Müşterilerinin Twit- ter Gönderilerinin Duygu Analizi. Çağ Üniversitesi Sosyal Bilimler Dergi- si, 18(1), 31-39. Bhatti, R. S., Kumar, P., & Kumar, D. (2010). A Fuzzy AHP Model for 3PL Selection in Lead Logistics Provider Scenarios. S. Parthasarathy (Edt.) Enterprise Information Systems and Implementing IT Infrastructures: Chal- lenges and Issues, 261-277. New York: Business Science Reference. Bilgi Teknolojileri ve İletişim Kurumu-Sektörel Araştırma ve Strateji Geliştir- me Dairesi Başkanlığı, (2011). Posta Sektörünün Düzenlenmesi Raporu. 1-59. Ankara. Burucuoğlu, M., & Yazar, E. E. (2020). Üçüncü Parti Platformda Kargo Fir- malarına Yapılan Müşteri Şikayetlerinin İçerik Analizi. Ekonomik ve Sosyal Araştırmalar Dergisi, 16(1), 99-114. Ding, J. F., Jhong, C. H., Huang, W. C., & Taleizadeh, A. A. (2014). Use of the Fuzzy AHP Method to Evaluate Key Factors Influencing New Cross-Strait Shuttle Shipping Routes. Marine Technology Society Jour- nal, 48(3), 125-137. Doğanalp, B. (2012).İnsan Kaynakları Seçme Sürecinde Bulanık Mantık Yakla- şımı: Görgül Bir Araştırma. (Doktora Tezi). Selçuk Üniversitesi, Sosyal Bilimler Enstitüsü: Konya Hirata, E., & Matsuda, T. (2023). Examining Logistics Developments in Post-Pandemic Japan Through Sentiment Analysis of Twitter data. Asian Transport Studies, 9, 100110, 1-7. Lazarevic, D., Dobrodolac, M., Švadlenka, L., & Stanivukovic, B. (2020). A model for Business Performance Improvement: A Case of The Postal Company. Journal of Business Economics and Management, 21(2), 564-592. Önden, İ., Eldemir F., & Çancı, M. (2015). Logistics Center Concept and Lo- cation Decision Criteria. Sigma, 33(3), 325-340. Pavic, Z., & Novoselac, V. (2013). Notes on TOPSIS Method. International Journal of Research in Engineering and Science (IJRES), 1(2), 5-12. PTT (2023). https://ptt.gov.tr/ (Erişim Tarihi: 25.12.2023). Putri, T. A., & Sari, N. (2021). Determination of Freight Transport Terminal Location by Using the AHP Method for Sustainable Urban Develop- ment. In IOP Conference Series: Earth and Environmental Science, 879, 1-7. Aslı Uğur Aydın Taştekin / Ayşegül Güdül 373 Rogerson C.M., Takis W. M., & Lenard T. M. (1993). Economies of Scale and Scope and Competition in Postal Services, M. A. Crew & P. R. Kleindor- fer, içinde, Regulation and the Nature of Postal and Delivery Services. (109-132), Boston: Springer. Sağır, H. & Doğanalp B. (2016). Bulanık Çok-Kriterli Karar Verme Perspekti- finden Türkiye İçin Enerji Kaynakları Değerlendirmesi. Kastamonu Üni- versitesi İktisadi ve İdari Bilimler Fakültesi Dergisi, 11, 233-256. 374 \| Posta, Haberleşme ve Kargo İşletmelerinin Web Sitelerinin TOPSIS Yöntemi... 1 Akdeniz Üniversitesi Hukuk Fakültesi, Deniz Hukuku Anabilim Dalı, myunusalhan@gmail.com, ORCID ID: 000-0002-6145-2591. Özet Kollektif şirket, 2011 tarihli 6102 sayılı Türk Ticaret Kanununun ‘‘Ticaret Şirketleri’’ başlıklı ikinci kitabında; tüzel kişiliği haiz bir ticaret şirketi olarak kabul edilmiştir. Bununla birlikte, kollektif şirketin, şahıs şirketi hüviyetinde olduğu da belirtilmiştir. Sahiden de TTK’da kollektif şirkete ilişkin düzenlemelerde (TTK m.211-303) şirketin tipik bir şahsi şirket niteliği arz ettiği anlaşılmaktadır. Zira kollektif şirket, gerçek kişiler arasında kurulan ve bütün ortakların sorumluluğu tam ve sınırsız olan bir yapıdadır. KAYNAKÇA Sağır, M. & Güdül, A. (2023). Evaluation of Child-Friendly Hotel: The Case of Side District. Beykoz Akademi Dergisi. 11(1). 393-406. Şenaran, L. E. (2023). E-ticarette Kargo Şirketi Seçiminde Etkili Olan Kriterle- rin AHP ile Karşılaştırılması: Otomotiv Yedek Parça Sektörü Uygulama- sı. (Yüksek Lisans Tezi). Karamanoğlu Mehmetbey Üniversitesi, Sosyal Bilimler Enstitüsü: Karaman. TDK (2023). https://sozluk.gov.tr/ (Erişim Tarihi: 25.12.2023). Ulutaş, A., & Karaköy, Ç. (2019). CRITIC ve ROV Yöntemleri ile Bir Kargo Firmasının 2011-2017 Yılları Sırasındaki Performansının Analiz Edilme- si. MANAS Sosyal Araştırmalar Dergisi, 8(1), 223-230. Uzun, H. (2013). Kargo Taşımacılık Sektöründe Hizmet Kalitesinin Değerlen- dirilmesi: AHP ve TOPSIS Yöntemi. Yüksek Lisans Tezi, Gazi Üniversi- tesi Eğitim Bilimleri Enstitüsü, Ankara. Bölüm 21 Kollektif Şirkette Ortaklığa Giriş Kollektif Şirkette Ortaklığa Giriş GİRİŞ Çalışmanın ana teması olan, kollektif şirkette ortaklığa giriş hususunda ise Kanunda kollektif şirket başlığı altında düzenlenmiş bir hüküm bulunmamaktadır. Şirkete yeni ortak alınması, pay devri ve ortağın ölümü halinde mirasçılarının şirkete katılımı türlerinde olmak üzere, şirkete katılımın üç farklı hali mümkündür. Bu hallerden, şirkete yeni ortak alımı ve payın devri suretiyle ortaklık sıfatının kazanımına ilişkin kanun koyucu, ‘‘Ticaret Şirketleri’’ başlıklı ikinci kitabın genel hükümler kısmında yer alan hüküm uyarınca (TTK m.126), Türk Borçlar Kanununda düzenlenen adi şirkete ilişkin hükümlere atıfta bulunmak suretiyle bu konudaki eksikliği gidermeye çalışmıştır. Mirasçıların şirkete katılımı konusu ise ‘‘Ortakların Şirketten Ayrılması’’ başlığı altında düzenlenmiş olup asıl olarak şirketten çıkma ve çıkarılmaya ilişkin olarak ifade edilmiştir. Ancak madde içeriğinde çıkma ve çıkarılma olmadan da şirketin mirasçılarla devam edebileceğine ilişkin ifadeler yer almıştır. Çalışmanın ‘‘Genel Olarak Kollektif Şirket’’ başlıklı birinci bölümünde, kollektif şirketin özellikleri üzerinde durularak kollektif şirket kavramı açıklığa kavuşturulmaya çalışılmış, daha sonra kollektif şirketin kuruluşuna ilişkin https://doi.org/10.58830/ozgur.pub401.c1769 375 376 Kollektif Şirkette Ortaklığa Giriş bilgiler verilmiş ve son olarak şirketin ehliyeti konusu üzerinde durulmuştur. Çalışmanın ‘‘Kollektif Şirkette Ortaklığa Giriş ve İç İlişki’’ başlıklı ikinci bölümünde, şirkete ortak olarak girilebilecek haller incelenmiş, daha sonra şirketin ortaklarla ve ortakların birbirleriyle olan ilişkileri, yani iç ilişkiler, detaylı olarak incelenmiştir. Böylelikle şirkette ortak sıfatıyla yer alan bir kişinin hangi hak ve yükümlülüklerin muhatabı olacağı ifade edilmeye çalışılmıştır. Çalışmanın ‘‘Kollektif Şirkette Ortaklığa Girişin Sonuçları’’ başlıklı üçüncü ve son bölümünde ise şirkete, kuruluştan sonra ortak olarak katılan kişilerin, hangi yolla ortaklık sıfatını iktisap etmişlerse ona göre ortaya çıkan sonuçlar üzerinde durulmuştur. 2 Meier – Hayoz A/ Fortmoser P, Schweizerisches Gesellschaftsrecht mit Einbezug des künftigen Rechnungslegungsrechts und der Aktienrechtsreform, Aufl. 11, 2012, s.2 (Naklen: Rıza Ayhan/ Hayrettin Çağlar/ Mehmet Özdamar, Şirketler Hukuku Genel Esaslar, 2. Baskı, Yetkin Yayınları, 2020, s. 15). 2 Meier – Hayoz A/ Fortmoser P, Schweizerisches Gesellschaftsrecht mit Einbezug des künftigen Rechnungslegungsrechts und der Aktienrechtsreform, Aufl. 11, 2012, s.2 (Naklen: Rıza Ayhan/ Hayrettin Çağlar/ Mehmet Özdamar, Şirketler Hukuku Genel Esaslar, 2. Baskı, Yetkin Yayınları, 2020, s. 15). 3 Tamer Bozkurt, Şirketler Hukuku, 9. Baskı, Legem Yayıncılık, 2018, s. 131. 4 Reha Poroy/ Ünal Tekinalp/ Ersin Çamoğlu, Ortaklıklar Hukuku – I, 14. Bası, Vedat Kitapçılık, 2019, s. 251; Ayhan/ Çağlar/ Özdamar, s. 43. 5 Ayhan/ Çağlar/ Özdamar, s. 198. 4 Reha Poroy/ Ünal Tekinalp/ Ersin Çamoğlu, Ortaklıklar Hukuku – I, 14. Bası, Vedat Kitapçılık, 2019, s. 251; Ayhan/ Çağlar/ Özdamar, s. 43. 3 Tamer Bozkurt, Şirketler Hukuku, 9. Baskı, Legem Yayıncılık, 2018, s. 131. 1.1. Kollektif Şirket Kavramı Özel hukuk bağlamında şirket, müşterek bir amacın elde edilmesi amacıyla bir sözleşmeye isitnaden kurulan kişi birliğidir2. Dolayısıyla şirketlerin en önemli özelliği kişi birliği olmalarıdır. Kollektif şirket ise ticari bir işletmeyi bir ticaret unvanı altında işletmek amacıyla, gerçek kişiler arasında kurulan ve ortaklarından hiçbirinin sorumluluğu şirket alacaklılarına karşı sınırlanmamış olan şirkettir (TTK m.211). Türk Ticaret Kanunu’nun ‘’Ticaret Şirketleri’’ başlıklı ikinci kitabında kollektif şirketin bir ticaret şirketi olduğu belirtilmiş (TTK m.124/1) ve nitelik olarak da şahıs şirketi olduğu ifade edilmiştir (TTK m.124/2). Kanunda şahıs şirketi olduğu belirtilen kollektif şirket, niteliği itibarıyla, TBK’da düzenlenen, gerçek kişilerden müteşekkil olan adi şirketlerin tüzel kişiliği haiz bir türü olarak kabul edilir3. Şahıs şirketlerinde en önemli şey kişi unsurudur. Zira bir şahıs şirketinin kuruluşunda sözleşmeye katılan kişiler birbirlerine duydukları güven neticesinde şirkete ortak olmaktadırlar4. Bu niteliği itibarıyla, şahıs şirketlerinin dolayısıyla kollektif şirketin de intiutu personae (karşı tarafın kişiliği göz önüne alınarak işlem yapılması) özelliğini haiz olduğu kabul edilmelidir5. 5 Ayhan/ Çağlar/ Özdamar, s. 198. Yunus Alhan \| 377 Yunus Alhan \| 377 377 Kollektif şirket ortaklarının TTK m.127’de ifade edilen değerleri sermaye olarak getirebileceği öngörülmüş ve bu da kollektif şirketleri iktisadi yönden avantajlı bir konuma getirmiştir. Ayrıca TTK m.218 hükmünde ortaklardan her birinin ayrı ayrı şirketi yönetme hak ve görevini haiz olduğu belirtilerek piyasa koşullarına kolayca uyum sağlayabilme olanağı da sağlanmıştır6. İfade edilen bu avantajlara karşın, şirket borçlarından dolayı ortakların sorumluluğunun sınırlandırılmamış olması (TTK m.236) ve ortaklardan birinin ölümü, ortaklardan birinin şirketteki tasfiye payının cebri icra yoluyla paraya çevrilmesi gibi durumlarda şirketin sona ermesi, kişileri kollektif şirket kurma konusunda kararsızlığa sevk etmektedir. Ülkemizde son yıllarda bu husus sayısal veriler ışığında net bir şekilde gözlemlenmektedir. 31.12.2016 tarihi itibarıyla tescilli kollektif şirket sayısı 12.744 iken, 31.12.2018 tarihi itibarıyla bu sayı 10.730’a düşmüştür7. Bu durum ilerleyen yıllarda da devam etmiş ve 2019 yılı sonu itibarıyla 14 kollektif şirket kurulmasına karşın 55 kollektif şirket kapanmış, 2020 yılı sonu itibarıyla 14 kollektif şirket kurulurken 60 tanesi kapanmış ve 2021 yılı sonu itibarıyla 24 şirket kurulmuşken 33 tanesi de kapanmıştır8. 1.2. Kollektif Şirketin Özellikleri Kanun koyucu tarafından TTK m.211’de düzenleme alanı bulan kollektif şirketin, söz konusu madde başlığının aksine tanımı yapılmamış aksine özellikleri belirtilmiştir. Ancak TTK m.211’de sayılanlara ek olarak kollektif şirketlerin tüzel kişiliği haiz olması da şirket yapısının özelliklerindendir (TTK m.125/1). 6 Ayhan/ Çağlar/ Özdamar, s. 198. 7 Ayhan/ Çağlar/ Özdamar, s. 199. 8 https://www.tobb.org.tr/BilgiErisimMudurlugu/Sayfalar/KurulanKapananSirketistatistikleri. php. 9 Rıza Ayhan/ Hayrettin Çağlar, Ticari İşletme Hukuku Genel Esaslar, 11. Bası, Yetkin Yayınları, 2018, s. 93. 9 Rıza Ayhan/ Hayrettin Çağlar, Ticari İşletme Hukuku Genel Esaslar, 11. Bası, Yetkin Yayınları, 2018, s. 93. 8 https://www.tobb.org.tr/BilgiErisimMudurlugu/Sayfalar/KurulanKapananSirketistatistikler php. 10 Ayhan/ Çağlar/ Özdamar, s. 199. 11 Ayhan/ Çağlar, s. 287. 12 Ayhan/ Çağlar, s. 292. 13 Ayhan/ Çağlar, s. 298. 14 Mehmet Bahtiyar, Ortaklıklar Hukuku, 13. Baskı, Adalet Yayınevi, 2019, s. 81; Bozkurt, s. 132. 1.2.2. Ticari İşletmenin Ticaret Unvanı Altında İşletilmesi Kanunda açık bir tanımı bulunmayan, ticari işletmenin devredilebilen malvarlığı haklarından olan11 ticaret unvanı, tacirlerin ticari işletmelerine dair iş ve işlemlerinde kullandıkları addır12. Kanun gereği tacir sayılan (TTK m.16/1) kollektif şirketler, tacir olmaları sebebiyle kanuna uygun bir ticaret unvanı seçmekle yükümlüdürler (TTK m.18/1). Kollektif şirketlerin ticaret unvanı, bütün ortakların veya ortaklardan en az birinin adı ve soyadıyla şirketi ve türünü gösterecek bir ibareyi içerir (TTK m.42/1, TSY m.43/5). Ticaret unvanının çekirdek kısmında ‘‘şirketi’’ ve ‘‘türünü’’ gösteren bir ibare bulunmalıdır13. Kollektif şirketin ticaret unvanına, işletmenin özelliklerini belirten veya unvanda yer alan kişilerin kimliklerini gösteren ya da hayali adlardan ibaret olan ekler yapılabilir (TTK m.46/1). 1.2.1.Ticari İşletme İşletmek Amacıyla Kurulması Kanun koyucu TTK m.211’de kollektif şirketlerin ancak ticari bir işletmeyi işletmek amacıyla kurulabileceğini belirtmiştir. Ticari işletme, esnaf işletmesi için öngörülen sınırı aşan düzeyde gelir sağlamayı hedef tutan, faaliyetlerin devamlı ve bağımsız şekilde yürütüldüğü işletmedir (TTK m.11/1). Ancak ticari işletme iktisadi bir değer olarak bir hukuk kişisi değildir9. Kollektif şirketin kuruluş döneminde şirket henüz faaliyete geçmediğinden, esnaf işletmesi faaliyetleri için öngörülen sınırı aşıp aşmayacağı belli değildir. Dolayısıyla şirket sözleşmesinde gösterilen ticari işletmenin hacmi ve Kollektif Şirkette Ortaklığa Giriş 378 öneminden ziyade ortakların amacına bakılmalı, ortakların müşterek amacı ticari bir işletmenin işletilmesi yönünde ise şirket, tür olarak kollektif şirket kabul edilmelidir10. 14 Mehmet Bahtiyar, Ortaklıklar Hukuku, 13. Baskı, Adalet Yayınevi, 2019, s. 81; Bozkurt, s. 132. 1.2.3. Ortaklarının Gerçek Kişi Olması Kollektif şirket, iki veya daha fazla gerçek kişinin bir araya gelerek kurduğu bir şirket türüdür (TTK m.211). Kanunda ortak sayısı bakımından herhangi bir üst sınır öngörülmemiştir. Fakat ortakların, şirketin borçlarından ikinci dereceden sorumlu olması ve yönetim ile temsile ilişkin hükümler dikkate alındığında fazla sayıda ortağın dahil olduğu bir sistemin kollektif şirket yapısına uygun olmadığı kabul edilmelidir14. Kanunda sınırlı ehliyetsiz kişilerin kollektif şirkete ortak olabileceğine dair bir hüküm bulunmamakla birlikte 4721 sayılı Türk Medeni Kanunu’nda düzenlenen, vesayet altındaki kişiye vesayet makamı tarafından bir meslek veya sanatın yürütülmesi için izin verilmiş ise, o kişi bununla ilgili her türlü olağan işlemleri yapmaya yetkilidir ve bu tür işlemlerden dolayı bütün malvarlığı ile sorumlu olacağına dair hüküm (TMK m.453), vesayet altındaki kişinin bir sanat veya meslekle uğraşması için vesayet makamının izninin gerektiğine dair hüküm (TMK m.462/7) ve bir işletmenin devralınması Yunus Alhan \| 379 Yunus Alhan \| 379 379 Yunus Alhan \| 379 Yunus Alhan Yunus Alhan veya tasfiyesi, kişisel sorumluluğu gerektiren bir ortaklığa girilmesi veya önemli bir sermaye ile bir şirkete ortak olunması için vesayet makamının izninden sonra denetim makamının da izninin gerektiğine (TMK m.463/3) dair hükümler birlikte değerlendirildiğinde, sınırlı ehliyetsizlerin (ayırt etme gücüne sahip küçükler ve kısıtlılar) kollektif şirkete ortak olabilecekleri kabul edilmektedir15. 1.2.4. Şirket Borçlarından Dolayı Ortakların Sorumluluğu Kollektif şirketin borç ve yükümlülükleri, şirketi temsile yetkili kişilerin, hukuki işlemlerle şirket tüzel kişiliği adına yaptıkları taahhütler, sebepsiz zenginleşme ve organların haksız fiilleri gibi kanundan doğan durumlardan oluşur16. Şirketin borç ve taahhütlerinden dolayı birinci dereceden şirket tüzel kişiliği sorumludur (TTK m.237/1, c.1). Ancak şirkete karşı yapılan icra takibi semeresiz kalmış veya şirket herhangi bir sebeple sona ermiş ise yalnız ortak veya ortakla birlikte şirket aleyhine dava açılabilir ve takip yapılabilir (TTK m.237/1, c.2). Dolayısıyla ortaklar, şirketin borç ve taahhütlerinden ikinci dereceden, müteselsil olarak ve bütün malvarlıklarıyla sorumludurlar (TTK m.236/1). Kollektif şirketlerde ortakların sorumluluklarının sınırlandırılması mümkündür. Ancak söz konusu sınırlandırma yalnızca ortaklar arasında hüküm ve sonuç doğurur. Çünkü TTK m.211 hükmü uyarınca ortaklardan hiçbirinin sorumluluğu şirket alacaklılarına karşı sınırlandırılamaz. Buna aykırı olarak sözleşmeye konan şartlar, üçüncü kişiler hakkında geçerli olmaz (TTK m.236/3). 15 Hasan Pulaşlı, Şirketler Hukuku Genel Esaslar, 5. Baskı, Adalet Yayınevi, 2017, s. 200. 16 Ayhan/ Çağlar/ Özdamar, s. 201. 17 Bozkurt, s. 133. 17 Bozkurt, s. 133. 1.3.1.1. Şirket sözleşmesinin şekli Kollektif şirket sözleşmesinin yazılı şekle tabi olduğunu belirten kanun koyucu (TTK m.212), ‘‘zorunlu kayıtlar’’ başlığı altında şirket sözleşmesinde bulunması mecburi hususları düzenleyerek (TTK m.213) aranan yazılı şeklin ‘‘nitelikli yazılı şekil’’ olduğunu kabul etmiştir. Sözleşmesi kanuna uygun şekilde yapılmayan veya TTK m.213’te zikredilen zorunlu kayıtlardan biri veya birkaçı eksik yahut geçersiz olan bir kollektif şirket adi şirket hükmündedir (TTK m.214/1). 1.3.1. Şirket Sözleşmesi Kollektif şirket sözleşmesi yazılı şekle tabidir; ayrıca sözleşmedeki imzaların noterce onaylanması veya şirket sözleşmesinin ticaret sicil müdürü yahut yardımcısı huzurunda imzalanması şarttır (TTK m.212). Kollektif şirketin kurulabilmesi için ortakların, katılma paylarını ortak bir amaca ulaşmak üzere bir araya getirmeleri, şirket sözleşmesinin de kollektif şirketin gerekli diğer özelliklerini ihtiva etmesi gerekir18. 1.2.5. Şirket Tüzel Kişiliği Kanunda ticaret şirketi olarak anılan, bir şahıs şirketi niteliğinde olan kollektif şirket, tüzel kişiliği haiz bir yapıdır (TTK m.125/1). Kollektif şirket, ticaret şirketi olması nedeniyle tacir sayılır (TTK m.16/1). Şirket, tüzel kişiliği haiz olduğu için, hak ve borçların ilk muhatabı olup, aktif ve pasif dava ehliyeti bulunmaktadır17 17 Bozkurt, s. 133. 380 Kollektif Şirkette Ortaklığa Giriş 1.3. Kollektif Şirketin Kuruluşu Kollektif şirketin kurulabilmesi için bir şirket sözleşmesinin yapılması ve şirketin ticaret siciline tescil edilmesi gerekir. 20 Abdulkerim Yıldırım, Borçlar Hukuku Genel Hükümler, 6. Baskı, Adalet Yayınevi, 2017, s. 94. 18 Ayhan/ Çağlar/ Özdamar, s. 202. 19 Ayhan/ Çağlar/ Özdamar, s. 203. 21 Poroy/ Tekinalp/ Çamoğlu, s. 217. 22 Ayhan/ Çağlar/ Özdamar, s. 204. 23 Ayhan/ Çağlar, s. 94. 24 Ayhan/ Çağlar/ Özdamar, s. 205; hükmün kabulü yönünde bakınız: Poroy/ Tekinalp/ Çamoğlu, s. 218. 1.3.1.2. Şirket sözleşmesinin içeriği Kollektif şirket sözleşmesine aşağıdaki kayıtların yazılması zorunludur (TTK m.213/1): Kollektif şirket sözleşmesine aşağıdaki kayıtların yazılması zorunludur (TTK m.213/1): a) Ortakların ad ve soyadlarıyla yerleşim yerleri ve vatandaşlıkları. Kollektif şirket ortaklarının Türk vatandaşı olması gerekmediği gibi Türkiye’de bir yerleşim yerlerinin olmasına da gerek yoktur19. a) Ortakların ad ve soyadlarıyla yerleşim yerleri ve vatandaşlıkları. Kollektif şirket ortaklarının Türk vatandaşı olması gerekmediği gibi Türkiye’de bir yerleşim yerlerinin olmasına da gerek yoktur19. b) Şirketin kollektif olduğu. Sözleşmelerin yorumunda geçerli olan güven teorisine göre, somut olay çerçevesinde tarafların gerçek ve ortak iradelerini ortaya koymak asıldır20. Bu durum Türk Borçlar Kanunu’nda da düzenleme alanı bulmuştur. Bir sözleşmenin türünün ve içeriğinin belirlenmesinde ve yorumlanmasında, tarafların yanlışlıkla veya gerçek amaçlarını gizlemek için kullandıkları sözcüklere bakılmaksızın, gerçek ve ortak iradeleri esas alınır (TBK m.19). Ancak kanun koyucu, kollektif şirketlere ilişkin hükümlerde, Yunus Alhan \| 38 381 Yunus Alhan şirketin kollektif şirket olarak kurulma amacının açıkça belirtilmesini istemiştir. c) Şirketin ticaret unvanı ve merkezi. Tüzel kişi tacir olan kollektif şirket, ticari işletmesine ilişkin işlemleri, ticaret unvanıyla yapmak ve işletmesiyle ilgili senetlerle diğer belgeleri bu unvan altında imzalamak zorundadır (TTK m.39/1). Kollektif şirket sözleşmesi yapılmaksızın, şirketin türünü gösterir bir kaydı içermese bile, ortak bir unvan altında, üçüncü kişilerle işlem yapılması veya onlara karşı haksız bir fiil işlenmesi halinde ortaklar üçüncü kişilere karşı müteselsilen sorumlu olurlar (TTK m.216/2). Sözleşmede şirketin merkezinin gösterilmesi de gerekir, aksi halde kollektif şirket adi şirket hükmünde olur (TTK m.214/1). Şirketin merkezi kavramından kasıt; şirket faaliyet ve ilişkilerinin yoğunlaştığı merkez olup şirketin adresi olarak anlaşılmamalıdır21. Şirket merkezinin değiştirilmesi için şirket sözleşmesi değişikliği gerekirken; adres değişikliği için böyle bir gereklilik söz konusu olmayacaktır22. d) Esaslı noktaları belirtilmiş ve tanımlanmış bir şekilde şirketin işletme konusu. Kanun koyucu tarafından ifade edildiği üzere kollektif şirket yalnızca ticari bir işletmeyi işletmek amacıyla kurulabilir (TTK m.211). Ticari bir işletmenin en önemli unsuru gelir sağlama amacıdır. Gelir sağlama hedefiyle yapılan bütün faaliyetlere de iktisadi faaliyet denir23. Dolayısıyla şirketin konusu iktisadi faaliyetlerden herhangi biri olabilir. d) Esaslı noktaları belirtilmiş ve tanımlanmış bir şekilde şirketin işletme konusu. Kanun koyucu tarafından ifade edildiği üzere kollektif şirket yalnızca ticari bir işletmeyi işletmek amacıyla kurulabilir (TTK m.211). Ticari bir işletmenin en önemli unsuru gelir sağlama amacıdır. Gelir sağlama hedefiyle yapılan bütün faaliyetlere de iktisadi faaliyet denir23. Dolayısıyla şirketin konusu iktisadi faaliyetlerden herhangi biri olabilir. 24 Ayhan/ Çağlar/ Özdamar, s. 205; hükmün kabulü yönünde bakınız: Poroy/ Tekinalp Çamoğlu, s. 218. 21 Poroy/ Tekinalp/ Çamoğlu, s. 217. 25 https://www.mevzuat.gov.tr/File/GeneratePdf?mevzuatNo=23093&mevzuatTur=Teblig& mevzuatTertip=5 26 Poroy/ Tekinalp/ Çamoğlu, s. 219; Ayhan/ Çağlar/ Özdamar, s. 206-207. 27 Ayhan/ Çağlar/ Özdamar, s. 207. 28 Poroy/ Tekinalp/ Çamoğlu, s. 219; Oruç Hami Şener, Teorik ve Uygulamalı Ortaklıklar Hukuku 1.3.2. Şirket Sözleşmesinin İmzalanması Kurucular tarafından imzalanan kollektif şirket sözleşmesinin noter tarafından onaylanması veya ticaret sicili müdürü ya da yardımcısı huzurunda imzalanması gerekmektedir (TTK m.212). Şirket sözleşmesinin ticaret sicili müdürlüklerinde imzalanması, ‘‘Şirket Kuruluş Sözleşmesinin Ticaret Sicili Müdürlüklerinde İmzalanması Hakkında Tebliğ25’’ dikkate alınarak yapılmalıdır. Sözleşmenin noterce onaylanması veya ticaret sicili müdürü ya da yardımcısı huzurunda imzalanması, imzaların kime ait olduğunun tasdiki niteliğindedir ve bu aşamada noter veya ticaret sicil müdürü ya da yardımcısı tarafından içerik denetimi yapılamaz26. Ticaret sicili müdürlüğünün içerik denetimi yapabilmesi tescil talebi ile birlikte mümkün olacaktır27. Kurucular, şirket sözleşmesinin noterlikçe onaylı bir suretini onay tarihinden itibaren onbeş gün içinde şirket merkezinin bulunduğu yerdeki ticaret siciline vererek şirketin tescilini istemek zorundadırlar (TTK m.215). 1.3.1.2. Şirket sözleşmesinin içeriği e) Her ortağın sermaye olarak koymayı taahhüt ettiği para miktarı; para niteliğinde olmayan sermayenin değeri ve bu değerin ne suretle biçilmiş olduğu; sermaye olarak kişisel emek konulmuşsa bu emeğin niteliği, kapsamı ve değeri. Kollektif şirkete katılma payı olarak konulabilecek değerler TTK m.127’de sayılan iktisadi değerlerdir. e) Her ortağın sermaye olarak koymayı taahhüt ettiği para miktarı; para niteliğinde olmayan sermayenin değeri ve bu değerin ne suretle biçilmiş olduğu; sermaye olarak kişisel emek konulmuşsa bu emeğin niteliği, kapsamı ve değeri. Kollektif şirkete katılma payı olarak konulabilecek değerler TTK m.127’de sayılan iktisadi değerlerdir. f) Şirketi temsile yetkili kimselerin ad ve soyadları, bunların yalnız başına mı, yoksa birlikte mi imza koymaya yetkili oldukları. Şirket sözleşmesinde buna ilişkin bir kayıt yoksa ortaklardan her biri ferdi olarak şirketi yönetme hakkını ve temsil yetkisini haiz sayılır (TTK m.218/1, TTK m.233/1). Dolayısıyla söz konusu kayıtların şirket sözleşmesinde yer almaması durumunda kollektif şirketin, TTK m.214/1 hükmü uyarınca adi şirket hükmünde olacağı tartışmalıdır24. 382 Kollektif Şirkette Ortaklığa Giriş Ortaklar emredici hükümlere aykırı olmamak kaydıyla, şirket sözleşmesine diledikleri kayıtları koyabilirler (TTK m.213/2). 25 https://www.mevzuat.gov.tr/File/GeneratePdf?mevzuatNo=23093&mevzuatTur=Teblig& mevzuatTertip=5 1.3.3. Ticaret Siciline Tescil ve İlan Ticaret siciline tescil, kural olarak istem üzerine yapılır (TTK m.27/1, TSY m.21/1). Tescil istemi merkezin bulunduğu yer ticaret siciline yazılı şekilde dilekçe ile veya elektronik olarak yapılır (TTK m.29/1, TSY m.23/1). Dilekçede istem açıkça belirtilir ve tescil edilecek olgular gösterilir. Dilekçeye doğrulayıcı belgelerin asıları ya da onaylı örnekleri eklenir (TSY m.23/2). Başvurunun elektronik ortamda yapılması halinde dilekçe ve belgeler güvenli elektronik imza ile imzalanır (TSY m.23/3). Dilekçe, tescil isteminde bulunmaya yetkili kişi veya kişilerce imzalanır (TSY m.23/4). Kollektif şirketin kuruluşunda kurucuların tamamı veya temsilcileri tescil başvurusunda bulunabilecek olup tescil başvurusunun tüm ortaklar veya temsilcileri tarafından imzalanması gerekir (TSY m.22/1, TSY m.22/2-ç). Tescili talep etmekten kaçınılması durumunda, ortaklardan her biri şirketin feshini isteyebileceği gibi şirket sözleşmesinin süresi içinde tescil edilmemesi halinde, sürenin sona ermesiyle birlikte dava yoluyla tescili isteyebilir28. Şirketin tescil ve ilanı yapılmamışsa, aradan ne kadar süre geçmiş 383 Yunus Alhan Yunus Alhan olursa olsun, ortaklardan herhangi birinin istemi üzerine ve bu ortağın noter aracılığıyla diğer ortaklara uygun bir süreyi içeren ihtar göndermiş olması şartıyla mahkemece feshe karar verilebilir (TTK m.243/1-d). Tescil ve ilanın üçüncü kişilere etkisi Kanunda düzenlenmiştir. Buna göre, ticaret sicili kayıtları nerede bulunurlarsa bulunsunlar, üçüncü kişiler hakkında, tescilin Türkiye Ticaret Sicili Gazetesinde ilan edildiği; ilanın tamamı aynı nüshada yayımlanmamış ise, son kısmının yayımlandığı günü izleyen iş gününden itibaren hukuki sonuçlarını doğurur (TTK m.36/1). Üçüncü kişilerin, kendilerine karşı sonuç doğurmaya başlayan sicil kayıtlarını bilmediklerine ilişkin iddiaları dinlenmez (TTK m.36/3). Tescili zorunlu olduğu hâlde tescil edilmemiş veya tescil edilip de ilanı zorunlu iken ilan olunmamış bir husus, ancak bunu bildikleri veya bilmeleri gerektiği ispat edildiği takdirde, üçüncü kişilere karşı ileri sürülebilir (TTK m.36/4). Kollektif şirket sicile tescille birlikte kurulur. Şirketin kuruluşunun, ticaret siciline ilanının yapılmamış olması şirketin tüzel kişiliğini kazanmasını engellemeyecektir zira ilan açıklayıcı etkiyi haizdir29. Ders Kitabı, 4. Bası, Seçkin Yayınevi, 2019, s. 216. 30 Poroy/ Tekinalp/ Çamoğlu, s. 217; Murat Alışkan, ‘‘İşletme Konusu ve 6102 Sayılı Türk Ticaret Kanununa Göre Ticaret Şirketlerinin Ehliyeti ve Temsili’’, MÜHFHAD, 18(2), 2012, s. 208. 31 Alışkan, s. 208. 29 Pulaşlı, s. 202. 30 Poroy/ Tekinalp/ Çamoğlu, s. 217; Murat Alışkan, ‘‘İşletme Konusu ve 6102 Sayılı Türk Ticare Kanununa Göre Ticaret Şirketlerinin Ehliyeti ve Temsili’’, MÜHFHAD, 18(2), 2012, s. 208. 31 Alışkan, s. 208. 2.1.1. Şirkete Yeni Ortak Alınması Kanunda, kollektif şirkette ortaklıktan çıkma ve çıkarılma halleri ile ilgili düzenleme mevcut iken; üçüncü bir kişinin kollektif şirkete ortak olarak alınmasına dair bir hüküm bulunmamaktadır. Kollektif şirkete ilişkin hüküm bulunmayan hallerde, ilk olarak Türk Medeni Kanununun tüzel kişilere ilişkin genel hükümleri (TMK m.47-55), bu kısımda hüküm bulunmayan hallerde ise Türk Borçlar Kanununun adi şirkete dair hükümleri (TBK m.620-649), şirket niteliğine uygun olduğu oranda uygulanır (TTK m.126). Netice itibarıyla, kollektif şirkete yeni ortak alınmasına ilişkin TTK’da hüküm bulunmadığından dolayı, konuya ilişkin olarak TBK m.632 hükmünün uygulanması gerekecektir. Buna göre, ortaklığa yeni bir ortak alınması, bütün ortakların rızasına bağlıdır (TBK m.632/1). Kollektif şirket sözleşmesinde ortakların kimliğine dair bilgilerin mevcut olması zorunlu tutulduğundan (TTK m.213/1-a), kollektif şirkete yeni ortak alınması durumunda şirket sözleşmesinin değiştirilmesi gerekecektir33. Kollektif şirkette, şirket sözleşmesinin değiştirilmesine ilişkin kararların ise oybirliğiyle alınması şarttır (TTK m.226/2). Değiştirilen sözleşmenin de yine ticaret siciline tescili gerekir34 2. KOLLEKTİF ŞİRKETTE ORTAKLIĞA GİRİŞ VE İÇ İLİŞKİ 2.1. Kollektif Şirkette Ortaklığa Giriş 33 Poroy/ Tekinalp/ Çamoğlu, s. 251; Ayhan/ Çağlar/ Özdamar, s. 238. 1.4. Kollektif Şirketin Ehliyeti Ticaret şirketlerinin ehliyetine ilişkin olarak 6762 sayılı TTK’da, ticaret şirketlerinin, şirket sözleşmesinde yazılı işletme konusuyla sınırlı olmak şartıyla bütün hakları iktisap ve borçları iltizam edebileceği ifade edilmiştir (m. 137). 6762 sayılı TTK’da yer alan bu düzenleme ile şirketler hukuku bakımından uzun yıllar kabul edilen bu ilkeye ‘‘ultra vires ilkesi’’ denmektedir. Bu ilke, bir ticaret şirketinin ehliyetinin, şirket sözleşmesinde yazılı olan işletme konusu ile sınırlı olmasını ifade eder30. Söz konusu ilke uyarınca, şirket sözleşmesinde yazılı işletme konusu sınırları içerisinde yer alan işlemler ‘‘intra vires’’, işletme konusu dışındaki işlemler ise ‘‘ultra vires’’ olarak adlandırılır. Ultra vires işlemler, yani şirket sözleşmesinde yer alan işletme konusu dışındaki işlemler yok hükmündedir31. 6102 sayılı TTK’nın 125. maddesi ile birlikte, 6762 sayılı TTK’da kabul edilen ultra vires ilkesi kaldırılmıştır. Anılan hüküm uyarınca, ticaret şirketleri, Türk Medeni Kanununun 48. maddesi çerçevesinde bütün haklardan yararlanabilir ve borçları üstlenenebilir (TTK m.125/2). Buna göre, tüzel kişiler, cins, yaş, hısımlık gibi yaradılış gereği insana özgü niteliklere bağlı 384 Kollektif Şirkette Ortaklığa Giriş olanlar dışındaki bütün haklara ve borçlara ehildirler (TMK m.48). Kanunda, esaslı noktaları belirtilmiş ve tanımlanmış bir şekilde şirketin işletme konusunun şirket sözleşmesinde belirtilmiş olmasının zorunlu olduğu belirtilmiştir (TTK m.213/1-d). Anılan hükümden, şirketin işletme konusu ile ilgili sınırlı ehliyeti haiz olduğu izlenimi olmasına karşın TTK m.125/2 açık hükmü karşısında, şirketin işletme konusu ile sınırlı ehliyet kabul edilmemeli, hak ehliyetiyle ilgili TMK m.48 hükmü esas alınmalıdır32. 32 Ayhan/ Çağlar/ Özdamar, s. 211. 34 Bozkurt, s. 154. 34 Bozkurt, s. 154. 35 Ahmet Ayar, ‘‘Adi Ortaklık Sözleşmesinde İç ve Dış İlişkiler’’, Yeditepe Üniversitesi Sosyal Bilimler Enstitüsü Özel Hukuk Ana Bilim Dalı (Doktora Tezi), 2015, s. 182. 36 Bahtiyar, s. 95. 37 Poroy/ Tekinalp/ Çamoğlu, s. 258. 38 Ayhan/ Çağlar/ Özdamar, s.120. 39 Ayhan/ Çağlar/ Özdamar, s.238. 2.1.2. Payın Devri Kanunda, ortaklık payının devredilmesi suretiyle kollektif şirkete yeni ortak katılımına ilişkin olarak bir düzenleme bulunmamaktadır. Dolayısıyla, yine TTK m.126 hükmü atfıyla TBK hükümlerine gidilmesi gerekecektir. Buna göre, ortaklardan biri tek taraflı olarak bir üçüncü kişiyi ortaklıktaki 34 Bozkurt, s. 154. 385 Yunus Alhan Yunus Alhan payına ortak eder veya payını ona devrederse, bu üçüncü kişi ortak sıfatını kazanamaz (TBK m.632/2). Hükümden anlaşılacağı üzere ortaklık payının kısmen veya tamamen devredilmesi mümkündür. Bununla birlikte, ortaklık payının kısmen veya tamamen devredilmesi, şirket sözleşmesinin değişikliğini gerektirdiğinden (TTK m.213/1-a, TTK m.226/2), ortaklık payının devrine tüm ortakların onay vermesi gerekir. Ortaklar, payın devrine ilişkin muvafakatleri zımni olarak da verebilirler35 Kollektif şirkette ortaklık payının devri iki ayrı işlemden oluşmaktadır. İlk olarak, payını devretmek isteyen ortakla, payı devralmak isteyen kişi arasında bir pay devir sözleşmesi yapılır36. Ortaklık payını devreden ile payı devralan arasında akdedilen devir sözleşmesi, şirket sözleşmesinin şekline tabidir37. Daha sonra ise, devrin hüküm ve sonuç doğurabilmesi için ortakların bu devir işlemine onay vermesi gerekir (TTK m.226/2). Ortaklar, devir işlemine, payın devri sözleşmesinden önce veya sonra onay verebilecekleri gibi; şirket sözleşmesine, payın serbestçe devredilebileceğine dair hüküm konularak da pay devrine önceden icazet verilebilecekleri kabul edilir38. Ortakların, devre onay vermemeleri halinde pay devirleri şirkete ve ortaklara karşı geçersiz olup, payı devreden ve payı devralan arasında akdedilen sözleşme, şirkete ve ortaklara karşı hüküm ifade etmez39. 37 Poroy/ Tekinalp/ Çamoğlu, s. 258. 35 Ahmet Ayar, ‘‘Adi Ortaklık Sözleşmesinde İç ve Dış İlişkiler’’, Yeditepe Üniversitesi Sosyal Bilimle Enstitüsü Özel Hukuk Ana Bilim Dalı (Doktora Tezi), 2015, s. 182. 2.1.3. Mirasçının Şirkete Katılımı Kollektif şirkette ortaklardan birinin ölümü halinde, şirket sözleşmesinde şirketin ölen ortağın mirasçılarıyla devam edeceğine ilişkin düzenleme yoksa, mirasçılarla diğer ortakların oybirliği ile verecekleri karar üzerine şirket bunların arasında devam edebilir. Mirasçılar veya içlerinden biri şirkette kalmaya razı olmazlarsa, diğer ortaklar, ölen ortağın razı olmayan mirasçılarına düşen payları ödeyerek onları şirketten çıkarır ve aralarında şirkete devam edebilirler. Bu durumda sağ kalan ortaklardan birinin şirketin devamına onay vermemesi sebebiyle oybirliği sağlanamadığı takdirde ise şirket sona erer (TTK m.253/1). Şirketin, ölen ortağın mirasçılarıyla diğer ortaklar arasında kollektif şirket olarak devam edeceği hakkında şirket sözleşmesinde hüküm varsa, mirasçılar kollektif sıfatıyla şirkete devam edip etmemekte serbesttirler. 386 Kollektif Şirkette Ortaklığa Giriş Mirasçılar şirketin devam etmesini isterlerse, diğer ortaklar bu isteği kabul etmek zorundadır. Ancak, kollektif sıfatıyla şirkette kalmak istemeyen mirasçı varsa, ölen ortağın payından kendisine düşen tutar ile komanditer olarak şirkete kabul edilmesini önerebilir. Diğer ortaklar bu öneriyi kabul etmek zorunda değildir. Mirasçılar şirkete kollektif ortak veya komanditer olarak girip girmeyeceklerini ortağın ölüm tarihinden itibaren üç ay içinde şirkete ihbar etmelidir. Durumun şirkete ihbarına kadar, mirasçılar şirkette komanditer olarak kalmış sayılırlar. Bu süre içinde ihbarda bulunmamış olan mirasçılar, sürenin sona ermesinden itibaren kollektif ortak sıfatını alırlar (TTK m.253/2). 2.2.1. Ortaklar Arasındaki İlişkilerin Mahiyeti Kollektif şirkette, ortakların birbirleriyle olan ilşkilerinin düzenlenmesinde sözleşme serbestisi geçerlidir (TTK m.217). Dolayısıyla kollektif şirkette iç ilişkiyi, şirketin ortaklarla ve ortakların birbirleriyle olan ilişkilerini, düzenleyen hükümler esas itibarıyla düzenleyici bir nitelik arz eder40. İç ilişkide, ortaklar arasında akdedilen sözleşme hükümlerinin uygulanması asıl olmasına rağmen, sözleşmenin sınırsız ve müteselsil sorumluluğa aykırı olan şartları üçüncü kişiler hakkında geçerli olmaz (TTK m.236). Kollektif şirkette iç ilişkide ortaklara serbesti tanınmasına karşın kanun koyucu bazı alanlarda müdahalelerde bulunma ihtiyacı hissetmiştir. Söz konusu düzenlemelerden birisi, herhangi bir ortağın denetim hakkının şirket sözleşmesiyle dahi kaldırılamayacak olmasıdır (TTK m.225). Kanun koyucunun müdahale ettiği alanlardan bir diğeri de, ortağa tanınmış olan hakları kaldıran veya daraltan sözleşme şartlarının geçersiz olduğuna ilişkin hükümdür (TTK m.228/2). 40 Ayhan/ Çağlar/ Özdamar, s. 212. 41 Ayar, s. 111. 41 Ayar, s. 111. 2.2.2. Sadakat ve Özen Yükümü Ortakların, taahhüt ettikleri sermaye koyma borcunu yerine getirmelerinden sonra da şirkete ve birbirlerine karşı sadakatle hareket etmeyi ve şirket menfaatlerini gözetmeyi sürdürmeleri beklenir41. Dolayısıyla kollektif şirketlerde ortakların iç ilişkilerde sadakat ve özen borcuna uyması gerekmektedir. Yunus Alhan \| 387 Yunus Alhan \| 387 387 Yunus Alhan \| Kollektif şirket, birbirine güvenen, birbirlerinin şahsi niteliklerini bilen ortaklar tarafından teşekkül eden bir birlik olduğu için ortaklar, diğer ortaklardan, kendilerinden beklenemeyecek bir özen göstermesini isteyemezler42. Dolayısıyla ortakların, diğer ortaklara ve şirkete karşı yükümlü olduğu sadakat ve özenin ölçüsü; ortağın kendi işlerinde gösterdiği olağan dikkat ve basiretten ibaret olmalıdır43. Adi şirkette de ortaklık işlerinde, her ortağın kendi işlerinde olduğu ölçüde çaba ve özen göstermekle yükümlü olduğu ifade edilmiştir (TBK m.628). 42 Ömer Faruk Öğütcü, ‘‘Adi Ortaklıkta İç İlişkiler’’, Hacettepe Üniversitesi Sosyal Bilimler Enstitüsü Özel Hukuk Ana Bilim Dalı (Yüksek Lisans Tezi), 2019, s. 55. 45 Ayhan/ Çağlar/ Özdamar, s. 213. 43 Ayar, s. 114; Ayhan/ Çağlar/ Özdamar, s.213. 44 Ayar, s. 77. 43 Ayar, s. 114; Ayhan/ Çağlar/ Özdamar, s.213. 2.2.3. Eşitlik İlkesi Kollektif şirkette iç ilişkilerde eşitlik ilkesinin uygulanması esastır. Ortakların, TTK m.127 gereğince şirkete koymayı taahhüt ettikleri katılma payları birbirinden farklı nitelikte olsa bile, her ortak aynı şartlar altında muamele görüp, aynı hak ve borçlara sahip olur44. Eşitlik ilkesi TTK’da açık bir şekilde düzenleme alanı bulmamıştır. Bu ilke asıl olarak, zımni bir şekilde, TBK’da ifade edilmiştir. Kollektif şirkete ilişkin kısımda eşitlik ilkesine dair hüküm bulunmadığı için TBK’da yer alan, adi şirkete ilişkin hükümler, şirket türünün niteliğine uygun olarak uygulama alanu bulacaktır (TTK m.126). Buna göre, sözleşmede aksi kararlaştırılmamışsa katılım payları, ortaklığın amacının gerektirdiği önem ve nitelikte ve birbirine eşit olmak zorundadır (TBK m.621/2). Eşitlik ilkesinin TTK’da da yansımaları mevcuttur. Söz konusu hükümler şunlardır; ortaklardan her birinin, ayrı ayrı şirketi yönetme hakkını ve görevini haiz olduğuna ilişkin düzenleme (TTK m.218/1) ve her ortağın bir oy hakkına sahip olduğu ve bunun kesinlikle değiştirilemez olduğuna ilişkin düzenlemedir (TTK m.226/1). Kollektif şirkette eşitlik ilkesi esas olmakla birlikte, ortaklar aralarında akdedecekleri bir sözleşmeyle (TTK m.217), iç ilişkide bazı ortaklara ek haklar verilebilir. Dolayısıyla sözleşmede aksine hüküm bulunmadıkça, her ortağın aynı şartlar altında eşit muameleye tabi tutulması asıldır45. 388 Kollektif Şirkette Ortaklığa Giriş 2.2.4. Ortaklık Hak ve Borçlarının Devri Şahıs şirketi olması nedeniyle, kollektif şirketlerde iç ilişkilerden kaynaklanan hak ve borçlar şahsa bağlıdırlar ve devredilemezler46. Ancak bu kuralın bazı istisnaları mevcuttur. Bu bağlamda TTK m.126’nın atfıyla, ortaklar ancak diğer ortakların rızasıyla şirketteki payını başkasına devredebilir (TBK m.632/1). Ortaklar ayrıca yönetim, denetim, oy ve temsil haklarını tümüyle devredemeseler de bazı durumlarda bu haklarını temsilci vasıtasıyla kullanabilmektedirler. Söz konusu hakların devamlı olarak devredilebilmesi yalnızca küçük ve kısıtlılar bakımından mümkündür47. Bir başka istisna ise katılma paylarına ilişkindir. Ortakların sermaye koyma borcu (TTK m.127- 128), şahsa bağlı bir ifa gerektirmiyorsa üçüncü bir kişi tarafından yerine getirilebilir (TBK m.83). Dolayısıyla TTK m .127/1’de düzenlenen, sermaye olarak getirilebilecek değerlerden, maddi edim niteliğinde olanlar üçüncü kişilerce ifa edilebilir48. 46 Furkan Derdiman, ‘‘Adi Ortaklık Yapısındaki Değişiklikler’’, İstanbul Üniveristesi Sosyal Bilimler Enstitüsü Özel Hukuku Ana Bilim Dalı (Yüksek Lisans Tezi), 2021, s. 118. 47 Ayar, s. 12. 48 Yıldırım, s. 253. 49 Pulaşlı, s. 206. 50 Ayhan/ Çağlar/ Özdamar, s. 218. 2.3. Ortaklar Arasındaki İlişkiler (İç İlişki) Kollektif şirketlerde iç ilişki kavramı, ortakların birbirleriyle ve şirketle olan ilişkilerini ifade etmek için kullanılmaktadır49. Kollektif şirkette, ortakların birbirleriyle olan ilişkilerinin düzenlenmesinde sözleşme serbestisi geçerlidir (TTK m.217). Söz konusu hüküm uyarınca, iç ilişkide sözleşme serbestisi geçerlidir. Ancak kanun koyucu tarafından bu kurala bazı istisnalar da getirilmiştir (TTK m.225, TTK m.228). İç ilişkiler, şahsi ilişkiler ve mali ilişkiler olmak üzere iki ana başlık altında sınıflandırılmıştır. 50 Ayhan/ Çağlar/ Özdamar, s. 218. 46 Furkan Derdiman, ‘‘Adi Ortaklık Yapısındaki Değişiklikler’’, İstanbul Üniveristesi Sosyal Bilimler Enstitüsü Özel Hukuku Ana Bilim Dalı (Yüksek Lisans Tezi), 2021, s. 118. 47 Ayar, s. 12. 47 Ayar, s. 12. 49 Pulaşlı, s. 206. 48 Yıldırım, s. 253. 51 Şener, s. 218; Öğütcü, s. 61. 52 Ayhan/ Çağlar/ Özdamar, s.219. 53 Şener, s. 219; Bahtiyar, s. 87. 54 Ayhan/ Çağlar/ Özdamar, s.219. 2.3.1.1. Yönetim hakkı Kanunda yönetim hakkı, ‘‘Şirketin Yönetimi’’ başlığı altında düzenlenmektedir (TTK m.218-224). Ticari bir işletmeyi bir ticaret unvanı altında işletmek amacıyla kurulan kollektif şirkette (TTK m.211), ortakların şirket tüzel kişiliği adına hareket etmesi ile işletme amacı gerçekleştirilebilir50. Kanunda ortaklardan her birinin ayrı ayrı şirketi yönetme hak ve görevine sahip olduğu belirtilerek (TTK m.218/1), kollektif şirkette ferdi yönetim 389 Yunus Alhan \| 389 Yunus Alhan esası kabul edilmiştir51. Anılan hüküm uyarınca şirketin yönetimi, ortaklar için hem hak hem de görevdir. Kanunda, yönetim işlerinin, şirket sözleşmesiyle veya çoğunluğun kararıyla ortaklardan birine veya birkaçına verilebileceği ifade edilmiştir (TTK m.218/1). Ancak Kanunda yönetici ortaklar ile diğer ortaklar arasındaki ilişkilerin ne tür bir hukuki niteliği haiz olduğu düzenlenmemiştir. Adi şirketlerde ise ortaklık sözleşmesinde aksine hüküm bulunmadıkça, yönetici ortaklar ile diğer ortaklar arasındaki ilişkilerin, vekâlet sözleşmesine ilişkin hükümlere tabi olduğu belirtilmiştir (TBK m.630/1). Ticaret şirketlerine ilişkin Kanunda hüküm bulunmayan hallerde, TBK’nın adi şirketlere dair hükümlerinin uygulanacağına ilişkin düzenleme (TTK m.126) mevcuttur. Ancak kollektif şirkette yönetim hakkının, konu bakımından vekalet ilişkisine kıyasla daha geniş bir kapsamda olduğu anlaşılmaktadır. Kollektif şirket yöneticilerinin, şirketin amacını ve konusunu elde etmek için yapılması gereken olağan işlem ve işleri organ sıfatıyla yaptığı ve vekalet ilişkisinde olduğu gibi müvekkilinden talimat almasının gerekmediği kabul edilmektedir52. Ancak, vekalet ilişkisine dair hükümlerin, şirketin niteliğine uygun düştüğü sürece, yöneticiler ile kollektif şirket arasındaki ilişkiler bakımından uygulanmasının (TTK m.126) sorun teşkil etmeyeceği ifade edilebilir. Yönetici ortak, diğer ortakların itiraz hakkı saklı kalmak kaydıyla, şirketin yönetimi kapsamında olağan işleri kendi görüşüne göre ve aksi belirtilmediği sürece, diğer yönetici ortakların katılımı olmaksızın bağımsız olarak yapar53. Zira kollektif şirkette ferdi yönetim esastır. Şirket sözleşmesinde hüküm bulunması halinde, sözleşmede öngörülen usule uygun olarak alınacak bir kararla, yönetici ortağa talimat verilebileceği de kabul edilmektedir54. Bunula birlikte, yönetim hakkı şirket sözleşmesiyle bir ortağa verilmişse, bu ortak, diğer ortaklar itiraz etseler ve karşı çıksalar bile, hileye dayalı olmamak şartıyla, şirketin yönetimi kapsamındaki hususlar için gereken işlemleri yapabilir (TTK m.222). Kanunda, kollektif şirkette yönetim hakkının kapsamı belirlenirken olağan ve olağanüstü işler ayrımı yapılmıştır. Buna göre, şirketin yönetimi kapsamındaki hususlar, şirketin amacını ve konusunu elde etmek için yapılması gereken olağan işlem ve işler ile sınırlıdır. Şirketi yönetenler, şirket menfaatine uygun gördükleri işlerde, olağan işlem ve işlerle sınırlı olmak şartıyla, sulh, feragat ve kabul ile tahkime de yetkilidirler. Olağan iş ve işlemler 390 Kollektif Şirkette Ortaklığa Giriş dışında kalan hususlarda ise ortakların oybirliği şarttır (TTK m.223). 55 Ayar, s. 171; Öğütcü, s. 78. 2.3.1.1. Yönetim hakkı Kanun koyucu söz konusu düzenlemede olağan işleri saymış ve bunların dışında kalan işlemlerin, sınırlı sayıda olmamak üzere, olağanüstü işler olduğuna ilişkin örnekler vermiştir. Buna göre, bağışta bulunmak, kefil olmak, üçüncü kişi lehine garanti vermek, ticari mümessil tayin etmek ve şirket konusuna girmiyorsa taşınmazları satmak, satın almak, teminat göstermek, şirketin özüne ilişkin üretim araçlarını elden çıkarmak, rehnetmek veya ticari işletme rehni kurmak gibi olağan iş ve işlemler dışında kalan hususlar olağanüstü işlemler olarak kabul edilip, bu hususlarda ortakların oybirliğinin bulunması şartı aranmıştır (TTK m.223). Yönetim yetkisi, şirket sözleşmesiyle verilmişse, bu yetkinin geri alınması veya sınırlandırılması şirket sözleşmesinin değişikliğini gerektirdiği için bu karar oybirliği ile alınmalıdır (TTK m.219, TTK m.226/2). Oybirliğinin sağlanamaması durumunda, haklı sebeplerin varlığında, ortaklardan birinin talebi üzerine, mahkeme kararı ile yönetim hak ve görevi sınırlandırılabilir veya kaldırılabilir (TTK m.219). Anılan hükümde haklı sebep olarak nitelendirilebilecek haller de ifade edilmiştir. Buna göre, görevin yerine getirilmesinde basiretsizlik, ağır ihmal veya yönetimde iktidarsızlık gibi durumlar haklı sebep sayılabilir (TTK m.219). Bunlara ek olarak, yöneticinin, yönetim kapsamına giren işlerde şirkete ihanet etmiş olması, kendisine düşen asli görevleri ve borçları yerine getirmemesi ve uğradığı sürekli bir hastalık veya diğer bir nedenden ötürü şirketin yönetimi kapsamına giren işleri yapmak için gerekli olan yeteneği ve ehliyetini kaybetmiş olması gibi durumlar da haklı sebep olarak nitelendirilebilir55. Yönetici, ortakların kararıyla atanmışsa, ortakların çoğunluğunun kararıyla görevden alınabilir. Çoğunluk elde edilemediği takdirde, yönetim işlerine bakan ortağın şirket sözleşmesini ihlal ettiği veya olayda haklı sebep bulunduğu iddiasıyla, her ortak ilgili yönetici ortağın görevden alınması için mahkemeye başvurabilir (TTK m.220). Şirket işlerinin yönetimi, ortakların tümüne veya birkaçına verilmiş ise, bunların her biri yalnız başına yönetim hak ve görevini haizdir. Bununla beraber, şirketi yönetmekle yükümlü olan ortaklardan bazısı, yapılacak bir işin, şirketin menfaatlerine uygun olmadığını ileri sürerlerse, yönetim hak ve görevini haiz diğer ortaklar, çoğunluk kararıyla o işi yapabilirler (TTK m.221/1). Şirket sözleşmesinde şirket işlerinin yönetimi kendilerine verilmiş ortakların birlikte hareket etmeleri yazılı ise, ortakların, gecikmesinde tehlike görülen hâller istisna olmak üzere, her işte anlaşmaları gerekir. 55 Ayar, s. 171; Öğütcü, s. 78. 391 Yunus Alhan \| 391 Yunus Alhan \| Anlaşamadıkları takdirde durum ortaklar kuruluna götürülür ve bu kurulca verilecek karara göre hareket edilir. Kollektif şirketin yönetimine dair işlerde ortakların itiraz hakkı mevcuttur. Yönetici sıfatını haiz olmayan ortakların itiraz hakkı olağanüstü işlemlerle sınırlıdır. Olağanüstü işlerde, kural olarak, bütün ortaklar yönetim hakkına sahip olduğu için (TTK m.223) bütün ortakların itiraz hakkının bulunduğu söylenebilir56. 2.3.1.1. Yönetim hakkı Yönetim şirket sözleşmesiyle bir ortağa verilmişse, bu ortak, diğer ortaklar itiraz etseler ve karşı çıksalar bile, hileye dayalı olmamak şartıyla, şirketin yönetimi için gereken işlemleri yapabilir (TTK m.222). Kanunda, itiraz hakkı herhangi bir şekle tabi tutulmamış olup işlem yapılana kadar itiraz edilebileceği (TBK m.625) ifade edilmiştir. Bununla birlikte, itiraz, yalnızca iç ilişkide hüküm ifade edecek olup yöneticinin itiraza rağmen işlemi gerçekleştirmesi halinde temsil yetkisi sınırları çerçevesinde şirket bu işlem ile bağlı olur57. Ortağa verilen yönetim yetkisi, şirketin sona erdiği tescil ve ilan edilinceye kadar devam eder (TTK m.251). Kural böyle olmakla birlikte yöneticinin ölümü, iflası veya kısıtlanması da yöneticilik sıfatını kendiliğinden58 sona erdirir (TTK m.243). 56 Bahtiyar, s. 88; Ayhan/ Çağlar/ Özdamar, s. 223. 57 Şener, s. 219. 58 Bahtiyar, s. 87. 59 Ayar, s. 154-155. 60 Şener, s. 225. 2.3.1.2. Oy hakkı Kollektif şirkette her ortak bir oy hakkına sahip olup bu kurala aykırı olan sözleşmeler geçersizdir (TTK m.226/1). Ek olarak, ortakların oy hakkı, nitelik itibarıyla yükümlülük olmayıp bir hak durumundadır. Kollektif şirkette oy hakkı, ortaklık sıfatının şahsa bağlı olmasından dolayı bizzat ortak tarafından kullanılır59. Bu kuralın istisnası ise tasfiye memurlarının atanmaları ve görevden alınmaları veya onlara verilecek talimatla ilgili kararlara katılma hakkının, bir ortağın iflasında iflas idaresine, ölümünde mirasçılara ve kısıtlanması durumunda kanuni temsilcisine ait olduğuna dair düzenlemedir (TTK m.268/2). Ortaklık sıfatına bağlı olarak tanınan, vazgeçilemez veya sınırlandırılamaz bir hüviyete sahip olan60 oy hakkı, bazı durumlarda kanun koyucu tarafından sınırlandırmaya tabi tutulmuştur. Buna göre, şirketi yönetmekle yükümlü olan ortaklardan bazısı, yapılacak bir işin, şirketin menfaatlerine uygun olmadığını 392 Kollektif Şirkette Ortaklığa Giriş ileri sürerlerse, yönetim hak ve görevini haiz diğer ortaklar, çoğunluk kararıyla o işi yapabilirler (TTK m.221/1). Ortağın oy hakkından yoksun tutulabileceğine dair bir diğer mesele, icra edilen işlemlerde ortağın kişisel menfaatinin bulunduğu durumlardır. Bu konuya ilişkin, kollektif şirketler çatısı altında mevcut bir düzenleme bulunmamaktadır. Bu durumda da anonim şirketlere ilişkin, pay sahibinin kendisi, eşi, alt ve üstsoyu veya bunların ortağı oldukları şahıs şirketleri ya da hâkimiyetleri altındaki sermaye şirketleri ile şirket arasındaki kişisel nitelikte bir işe veya işleme veya herhangi bir yargı kurumu ya da hakemdeki davaya ilişkin olan müzakerelerde oy kullanamayacağına dair hüküm (TTK m.436/1) veya dernek üyelerinin, dernek ile kendisi, eşi, üstsoyu ve altsoyu arasındaki bir hukukî işlem veya uyuşmazlık konusunda alınması gereken kararlarda oy kullanamayacağına ilişkin düzenlemenin (TMK m.82) kıyas yoluyla uygulanması gündeme gelebilecektir61. Kollektif şirkette ortaklar, oylarını diğer bütün ortaklara hitaben açıklar62. Ortaklar açıklamış oldukları oyları, karar alınıncaya dek, herhangi bir sebep göstermeksizin değiştirebilir ve irade sakatlığı hallerine dayanarak verdikleri oyun iptalini de talep edebilirler63. 61 Ayhan/ Çağlar/ Özdamar, s. 235. 62 Ayhan/ Çağlar/ Özdamar, s. 235. 63 Şener, s. 225. 64 Öğütcü, s. 43. 65 Bahtiyar, s. 89. 2.3.1.3. Denetim ve bilgi edinme hakkı Kollektif şirkette, yönetim hakkını ve görevini haiz olmasa bile her bir ortak şirket işlerinin gidişi hakkında bizzat bilgi edinmek, şirketin belgelerini ve defterlerini incelemek, bunlara göre kendisi için şirketin finansal durumunu gösterecek bir hesap tablosu düzenlemek hakkını haizdir. Buna aykırı olan sözleşmeler ise geçersizdir (TTK m.225). Ortaklık sıfatına bağlı olan, dolayısıyla da şahsi bir nitelik arz eden denetleme ve bilgi edinme hakkı; yönetici ortağa, şirket tasfiye halindeyse tasfiye memuruna yöneltilerek kullanılabilir (TTK m.301/1). Şahsa bağlı bir niteliği haiz olması nedeniyle denetleme ve bilgi edinme hakkının, diğer ortakların onayı olmadan üçüncü bir şahsa devri mümkün değildir64. Bununla birlikte denetim hakkı ortağa pasif bir yetki verir dolayısıyla da ortağa bu hak yoluyla doğrudan müdahalede bulunma salahiyeti vermez65. Denetim ve bilgi edinme hakkı ortaklık sıfatına bağlı bir hak olup bu sıfatın sona ermesiyle söz konusu hak da ortadan kalkar. Ancak kanun koyucu tarafından bu duruma bir istisna getirilmiştir. Buna göre, çıkarılan veya çıkan Yunus Alhan \| 39 393 Yunus Alhan \| Yunus Alhan ortak, evvelce başlanmış işlerin hemen tasfiyesi mümkün olamadığı takdirde, her faaliyet dönemi sonunda, o yıl içinde bitirilen işlerin hesaplarını ve devam etmekte olan işlemlerin o tarihteki durumu hakkında bilgi verilmesini isteyebilir (TTK m.263/2). 66 Ayar, s. 116. 67 Ayhan/ Çağlar/ Özdamar, s. 225. 68 Ayar, s. 117. 2.3.2. Mali Nitelikteki İlişkiler 2.3.2. Mali Nitelikteki İlişkiler 69 Poroy/ Tekinalp/ Çamoğlu, s. 213. 2.3.1.4. Rekabet yasağı Kollektif şirkette ortakların, sözleşmede belirtilen amacın gerçekleşmesine yönelik olarak olumsuz davranışlardan kaçınması ve böylelikle sadakat yükümlülüğüne uygun hareket etmeleri beklenir66. Buradan hareketle rekabet yasağı bir başlık olarak kanun koyucu tarafından düzenleme alanı bulmuştur. Kanunda, bir ortağın, ortağı olduğu şirketin yaptığı ticari işler türünden bir işi, diğer ortakların izni olmaksızın kendi veya başkası hesabına yapamayacağı gibi aynı tür ticari işlerle uğraşan bir şirkete sorumluluğu sınırlandırılmamış ortak olarak giremeyeceği ifade edilmiştir (TTK m.230/1). Hükmün devamında ise yeni kurulan bir şirkete giren ortağın, daha önce kurulmuş diğer bir şirketin de sorumluluğu sınırlandırılmamış ortaklarından olduğunu diğer ortaklar bildikleri hâlde önceki şirketten ilişiğinin kesilmesini aralarında açıkça kararlaştırmamışlarsa, bu durumu kabul ettiklerinin varsayılacağı belirtilmiştir (TTK m.230/2). Benzer bir şekilde, rekabet yasağına aykırı davranışın diğer ortaklar tarafından bilinmesine rağmen sessiz kalınması, izin verildiği anlamına gelecektir67. Rekabet yasağına aykırılık halinde, şirket, bu ortaktan tazminat istemekte veya tazminat yerine bu ortağın kendi adına yaptığı işleri şirket adına yapılmış saymakta, üçüncü kişilerin hesabına yapmış olduğu işlerden doğan menfaatlerin şirkete bırakılmasını istemekte serbesttir (TTK m.231/1). Hükümden anlaşılacağı üzere, rekabet yasağı emredici bir mahiyette değildir. Dolayısıyla ortaklar, ortağı olduğu şirketin yaptığı ticari işler türünden bir işi, diğer ortakların izni olmaksızın kendi veya başkası hesabına yapamayacağı gibi aynı tür ticari işlerle uğraşan bir şirkete sorumluluğu sınırlandırılmamış ortak olarak girmesine izin verebilirler. Ortakların vereceği bu izin, bir karara istinaden verilebileceği gibi örtülü de olabilir68. Rekabet yasağına aykırılık halinde, TTK m.231/1 hükmünde belirtilen yaptırımlardan hangisinin uygulanacağına diğer ortaklar çoğunlukla karar verir (TTK m.231/2). Rekabet yasağına aykırı olarak icra edilen iş ve işlemlerden dolayı hakları ihlal edilen ortaklar şirketin feshini isteyebilecekleri gibi (TTK m.231/3) ortağın şirketten çıkarılmasına karar verebilirler (TTK m.255/1). 394 Kollektif Şirkette Ortaklığa Giriş Kanunda ifade edilen yaptırımları (TTK m.231/1) uygulayabilme hakkının, rekabet yasağına aykırı işlemin yapıldığının veya ortağın diğer bir şirkete girdiğinin öğrenildiği tarihten başlayarak üç ay ve her halde işlemin yapıldığı tarihten itibaren bir yıl sonra zamanaşımına uğrayacağı kabul edilmiştir (TTK m.231/2). 2.3.2.1. Sermaye koyma borcu Kanunda, sermaye şirketlerinin aksine, kollektif şirketin kurulabilmesi için asgari bir sermaye miktarı öngörülmemiştir. Dolayısıyla, kollektif şirketin bir ticari işletmeyi işletmek amacıyla kurulabileceği (TTK m.211) dikkate alındığında, kollektif şirket sermayesinin, ticari bir işletmeyi işletebilecek kapasitede olması gerektiği ifade edilebilir69. Kollektif şirket sözleşmesinde, her ortağın sermaye olarak koymayı taahhüt ettiği para miktarı; para niteliğinde olmayan sermayenin değeri ve bu değerin ne suretle biçilmiş olduğu; sermaye olarak kişisel emek konulmuşsa bu emeğin niteliği, kapsamı ve değerinin gösterilmesi zorunludur (TTK m.213/1-e). Bu kapsamda, şirketin amacını gerçekleştirmeye yönelik olarak şu unsurlar sermaye olarak konulabilir (TTK m.127/1): - Para, alacak, kıymetli evrak ve sermaye şirketlerine ait paylar (TTK m.127/1-a). - Fikrî mülkiyet hakları (TTK m.127/1-b). - Taşınırlar ve her çeşit taşınmaz (TTK m.127/1-c). - Taşınır ve taşınmazların faydalanma ve kullanma hakları (TTK m.127/1-d). - Kişisel emek (TTK m.127/1-e). - Ticari itibar (TTK m.127/1-f). - Ticari işletmeler (TTK m.127/1-g). - Haklı olarak kullanılan devredilebilir elektronik ortamlar, alanlar, adlar ve işaretler gibi değerler (TTK m.127/1-h). - Haklı olarak kullanılan devredilebilir elektronik ortamlar, alanlar, adlar ve işaretler gibi değerler (TTK m.127/1-h). - Maden ruhsatnameleri ve bunun gibi ekonomik değeri olan diğer haklar (TTK m.127/1-i). - Devrolunabilen ve nakden değerlendirilebilen her türlü değer (TTK m.127/1-j). - Devrolunabilen ve nakden değerlendirilebilen her türlü değer (TTK m.127/1-j). 69 Poroy/ Tekinalp/ Çamoğlu, s. 213. 395 Yunus Alhan Yunus Alhan Bir kollektif şirketten bahsedebilmek için her ortağın, şirketin amacını gerçekleştirmeye yönelik bir katılma payı ile şirkete katılması gerekir70. Usulüne göre düzenlenmiş ve imzalanmış şirket sözleşmesiyle kararlaştırılan katılma payı taahhütleri, ortakların sermaye borcunu oluşturur (TTK m.128/1). Şirket sözleşmesinde aksine hüküm bulunmadıkça, sermaye olarak konan ayınların mülkiyeti şirkete ait ve haklar şirkete devredilmiş olur (TTK m.131/2). Şirket, her ortağın sermaye koyma borcunu yerine getirmesini isteyebileceği ve dava edebileceği gibi, yerine getirmede gecikme sebebiyle uğradığı zararın tazminini de isteyebilir. Tazminat istemi için ihtar şarttır. Kollektif şirkette bu davayı ortaklar da açabilir (TTK m.128/7). 70 Ayhan/ Çağlar/ Özdamar, s. 215. 70 Ayhan/ Çağlar/ Özdamar, s. 215. 71 Bahtiyar, s. 85. 71 Bahtiyar, s. 85. 72 Bahtiyar, s. 86. 73 Ayhan/ Çağlar/ Özdamar, s. 217. 74 Derdiman, s. 208-209. 75 Ayhan/ Çağlar/ Özdamar, s. 217. 76 Ayhan/ Çağlar/ Özdamar, s. 217. 2.3.2.3. Tasfiye payı – Ayrılma payı Sona ermeyle birlikte, şirket tasfiye sürecine girer. Tasfiye işlemi ile şirketin net malvarlığı ortaya çıkarılır72. Şirketin net varlığı, şirket sözleşmesine veya sonradan verilecek karara göre, tasfiye memurları tarafından dağıtılır. Sözleşmede aksine hüküm veya ortakların kararı bulunmadığı takdirde dağıtma para olarak yapılır (TTK m.300). Ortakların, dağıtılacak tasfiye payına iştirak hakkı vardır. Söz konusu hak şarta bağlı bir mali hak olup73, şirketin tasfiyesi tamamlanmadan talep edilemez. Ortaklardan birinin şirketten çıkması veya çıkarılması durumunda ise o ortağa tasfiye payı yerine ayrılma payının verilmesi gerekir. Ayrılma payı, ayrılma tarihine en yakın olan tarih esas alınarak belirlenecek bilançoya dayanılarak (TTK m.260), farazi olarak yapılacak bir tasfiyeye göre belirlenir74. Tıpkı tasfiye payı gibi; ayrılma payı da şarta bağlı mali bir hak olup75, ortağın şirketle ilişiği kesilmeden talep edilemez. Tasfiye payı tespit edildikten sonra, ilk olarak, ortakların sermaye olarak koydukları katılım payları kendilerine verilir. Katılım payı olarak bir malın kullanma hakkı devredilmiş ise söz konusu mal iade edilir ancak kullanma hakkının karşılığı olarak bir şey verilmez76. 2.3.2.2. Kâr payı hakkı ve zarara katılma Her ortak, şirketten faaliyet dönemi sonunda gerçekleşen kârdan kendisine düşen payı, kanuna veya şirket sözleşmesine göre yılsonu bilançosu çıkarılmamışsa çıkarılmasını, bilançoda kâr payı saptanmamışsa saptanmasını ve alacaklarını istemek hakkına sahiptir (TTK m.228/1). Zarar payı ile ilgili olarak ise ortaklar oybirliği ile karar almadıkça hiçbir ortak, sermayesinden eksilen kısmı tamamlamaya zorlanamaz (TTK m.229/1). Sermayenin zararla eksilen kısmı, aksine karar yoksa, gerçekleşecek kâr ile kapatılır (TTK m.229/2). Zararla ilgili TTK m.229 hükmü saklı kalmak kaydıyla ortaklar, kâr ve zarardan kendilerine düşen payın belirlenmesini, şirket sözleşmesiyle veya sonradan alacakları bir kararla, içlerinden birine veya bir üçüncü kişiye bırakabilirler (TTK m.227/2). Kâr ve zarara katılmaya ilişkin şirket sözleşmesinde hüküm bulunması halinde, ortakların kâr ve zarara katılması bu hükümlere göre yapılır. Meğer ki sözleşmede öngörülen oran ‘‘aslan payı’’ niteliğinde olmasın71. Sözleşmede hüküm bulunmaması halinde, her ortağın kâr ve zarardaki payı, katılım payının değerine ve niteliğine bakılmaksızın eşittir (TTK m.227/3, TBK m.623/1). Sözleşmede ortakların kâr veya zarara katılım paylarından biri belirlenmişse bu belirleme, diğerindeki payı da ifade eder (TBK m.623/2). Şirketin faaliyet dönemi sonunda, finansal tabloların onaylandığı toplantıda kârın dağıtımı da karara bağlanır. Ortaklar, bu kararın kanuna, şirket sözleşmesine, şirket kararlarına veya dürüstlük kuralına aykırı olması hâlinde, kârın kullanılması hakkındaki karar tarihinden itibaren üç ay içinde iptal davası açabilirler (TTK m.227/1). Ortakların, kâr ve zarardan kendilerine 71 Bahtiyar, s. 85. 396 Kollektif Şirkette Ortaklığa Giriş düşen payın belirlenmesini, şirket sözleşmesiyle veya sonradan alacakları bir kararla, içlerinden birine veya bir üçüncü kişiye bırakabileceklerini ifade etmiştik. Bu ortağın veya üçüncü kişinin vereceği kararın hakkaniyete aykırı olması durumunda ise söz konusu kararın öğrenilmesinden itibaren üç ayın geçmesi, belirlenen kâr payının ortak tarafından tamamen veya kısmen alınması veya başka bir kimseye devredilmesi, zararın ödenmesine başlanması gibi açık veya zımni kabulü gösteren durumlarda dava hakkı düşecektir (TTK m.227/2). 2.3.2.4 Ücret ve faiz isteme ve masrafı talep etme hakkı Kanunlarda aksine hüküm yoksa, şirket sözleşmesiyle ortakların, şirketteki hizmetleri sebebiyle kendilerine ücret verilmesi kabul olunabilir (TTK m.132). Her ortak, şirket sözleşmesinde hüküm bulunması halinde, hak ettiği ücreti istemek hakkına sahiptir (TTK m.228/1). Ayrıca, ortağın yapmış olduğu hizmet, şirket sözleşmesinde ortaklara yüklenen ödevlerden 397 Yunus Alhan Yunus Alhan değil ise hizmet, vekalet veya istisna sözleşmesi hükümleri uyarınca, üçüncü bir kişi gibi ücret talep edilebilir77. Şirketin iflası halinde ise ortaklar, işlemiş faizler, ücretler ve şirket lehine yaptıkları giderler için herhangi bir alacaklı gibi masaya girebilirler (TTK m.241). Kollektif şirkette ortakların, şirket sözleşmesinde hüküm bulunması halinde, koydukları sermayeler için faiz verilmesi kabul edilebilir (TTK m.132). faizin verilebilmesi için şirketin faaliyet dönemi sonunda kar etmiş olması gerekmez78. Ortakların, şirket sözleşmesinde öngörülen ücreti talep etme haklarını ve koydukları sermaye payının faizini talep etme haklarını kaldıan veya sınırlandıran sözleşme şartları geçersiz olup söz konusu haklar emredici niteliktedir (TTK m.228/2). Ortakların şirket işleri için yapmış oldukları giderler, şirket tarafından ödenir79. Gerek şirket yöneticileri gerekse yönetici sıfatı olmayan ortaklar olsun, şirket işleri için emek sarfeden ortağa, şirket tarafından ödeme yapılması gerekir (TBK m.627/3). Kollektif şirkette ortaklardan her biri, şirkete ödünç olarak verdiği parayı ve kararlaştırılmışsa faizini de talep etme hakkına sahiptir (TTK m.228/1). 77 Bahtiyar, s. 85. 78 Ayhan/ Çağlar/ Özdamar, s. 218. 79 Ayhan/ Çağlar/ Özdamar, s. 218. 80 Poroy/ Tekinalp/ Çamoğlu, s. 253. 80 Poroy/ Tekinalp/ Çamoğlu, s. 253. 3.2. Pay Devri Sonucu Ortaklığa Girmenin Sonuçları Kollektif şirkette ortaklık payının devredilmesi, tıpkı şirkete yeni ortak alınmasında olduğu gibi, ortaklık sıfatına bağlı bazı hak ve borçların devri sonucunu da doğurur. şirkete giren yeni ortak, ayrılan ortağın bitin hak ve borçlarının tarafı haline gelir. Ortaklık sıfatına bağlı olarak, yönetim, oy, denetim ve bilgi edinme gibi haklardan istifade edebilecekken; sermaye koyma, zarara katılma ve rekabet yasağına uygun davranma gibi yükümlülüklere de katlanmak durumundadır. Payın devredilmesi suretiyle şirkete ortak olacak kişinin devraldığı ortaklık payında taşınmaz bir mal bulunması durumunda, devir sözleşmesinin resmi olarak yapılması gerekmeyeceği gibi; resmi şekle tabi ayrı bir sözleşmenin de akdedilmesi gerekmez. Çünkü burada devre konu olan söz konusu taşınmaz mal olmayıp, ortaklığın malvarlığına ilişkin hak sahipliğinin devridir81. Şirket borçları bakımından ise şirkete yeni giren kişi, girme tarihinden evvel doğmuş olsa bile, şirketin borçlarından ve taahhütlerinden diğer ortaklarla birlikte müteselsilen ve bütün malvarlığı ile sorumludur (TTK m.236/2). Ortaklar, bunun aksini sözleşmede kararlaştırabilirler. Ancak bu şart, iç ilişkide geçerli olup, üçüncü kişilere karşı hüküm ifade etmez (TTK m.236/3). 3.1. Şirkete Yeni Ortak Alınmasının Sonuçları Kollektif şirkete yeni bir ortağın dahil olmasıyla birlikte, tüm ortakların oybirliğiyle değiştirilen şirket sözleşmesine (TTK m.226/2), yeni giren ortağın adı, soyadı, yerleşim yeri ve vatandaşlığı hususları eklenir (TTK m.213/1-a) ve bu durum ticaret siciline tescil ve ilan olunur (TTK m.215). Şirkete yeni giren ortağa ilişkin söz konusu kayıtlar sözleşmede yer almazsa kollektif şirket adi şirket hükmünde olur ve şirket hakkında, TBK’da düzenleme alanı bulan, adi şirketlere ilişkin hükümler uygulanır (TTK m.214/1). Şirkete yeni katılan ortak, ortaklık payından doğan hak ve borçları, aktif ve pasifleriyle devralmış olur80. Yeni ortak, pay sahipliğinden kaynaklanan yönetim (TTK m.218), oy (TTK m.226), denetim ve bilgi edinme (TTK m.225), kâr payı (TTK m.227) ve ücret (TTK m.228) talep etmek gibi 398 Kollektif Şirkette Ortaklığa Giriş hakları kullanabilecekken; aynı zamanda sermaye koyma (TK m.127), zarara katılma (TTK m.227) ve rekabet yasağına uygun davranma (TTK m.230) gibi yükümlülüklere de riayet etmelidir. Şirkete yeni giren kişi, girme tarihinden evvel doğmuş olsa bile, şirketin borçlarından ve taahhütlerinden diğer ortaklarla birlikte müteselsilen ve bütün malvarlığı ile sorumlu olur (TTK m.236/2). Bu hükme aykırı olarak sözleşmeye konan şartlar üçüncü kişiler hakkında geçerli olmayıp, iç ilişkide hüküm ifade eder (TTK m.236/3). 81 Ayar, s. 181. 82 Poroy/ Tekinalp/ Çamoğlu, s. 258. 83 Poroy/ Tekinalp/ Çamoğlu, s. 258. 82 Poroy/ Tekinalp/ Çamoğlu, s. 258. y/ p/ Ç ğ , 83 Poroy/ Tekinalp/ Çamoğlu, s. 258. 84 Poroy/ Tekinalp/ Çamoğlu, s. 258. 85 Poroy/ Tekinalp/ Çamoğlu, s. 260. 84 Poroy/ Tekinalp/ Çamoğlu, s. 258. 3.3. Mirasçının Şirkete Katılımının Sonuçları Ölen ortağın mirasçılarının ortaklığa karşı hakları, murisin tasfiye payına ilişkin talep hakkından oluşur82. Mirasçılar şirkete karşı, miras payının alacaklısı konumunda bulunan üçüncü kişi durumundadırlar83. Mirasçının şirkete katılımının sonuçlarını iki ayrı başlık altında incelemek gerekmektedir. İlk durum, ölen ortağın mirasçılarıyla diğer ortaklar arasında ortaklığın devam edeceğine dair şirket sözleşmesinde hüküm bulunmaması 399 Yunus Alhan Yunus Alhan halidir. Bu durumda mirasçıların her biri şirkete katılıp katılmama hususunda serbesttirler. Mirasçılar bu konudaki isteklerini şirkete değil, bizzat ortaklara bildirmelidir84. Dolayısıyla bu durumda, mirasçıların şirkete ortak sıfatıyla girmesi, şirkete yeni ortak alınması prosedüründen farklılık arz etmeyecektir. Böylelikle, mirasçıların şirkete ortak sıfatıyla girebilmesi, ortaklar ile aralarında yapacakları ve şirket sözleşmesinin değişikliğini gerektirdiği için oybirliğiyle kabul edilmesi gereken bir sözleşme sayesinde mümkün olacaktır. Şirkete girmek istemeyen veya ortaklar ile anlaşamayan mirasçılara ise miras payları ödenir ve bunlar şirketten çıkarılır. Kanunun 243.maddesi uyarınca, ortaklığın mirasçılarla sürdürülmesi hususunda şirket sözleşmesinde hüküm yoksa, ortaklardan birinin ölümü şirketin sona ermesine neden olur. Buradan hareketle belirtmek gerekir ki mirasçılardan hiçbiri şirkete katılmak istemezse bu durumda ortakların şirketi kendi aralarında devam ettirme hususunda oybirliğiyle bir karar almaları gerekir. İki kişilik kollektif şirketlerde ise ölen ortağın mirasçıları şirkete girmeyi kabul etmezlerse şirket sona erer. İkinci durum ise ölen ortağın mirasçılarıyla ortaklığın devam edeceğine ilişkin şirket sözleşmesinde bir düzenleme bulunması halidir. Sözleşmeye konulan bu kayıt ile ölüm halinde ortaklığın sona ermesini önlemek amaçlanmıştır. Bu durumda mirasçılar, şirket sözleşmesinde yer alan hükme dayanarak ortak sıfatıyla şirkete katılabilirler. Mirasçıların bu talebini diğer ortaklar kabul etmek zorundadırlar. Dolayısıyla söz konusu talep ortakların kabulüne bağlı olmayan kurucu yenilik doğuran bir hakkın kullanılması mahiyetinde olur85.Ortağın ölümü sonucunda mirasçıların şirkete girmesi sözleşme değişikliğini gerektirdiği için bu durumun ticaret siciline tescil ve ilanı gereklidir. Kanunun 216.madde hükmü dikkate alındığında, yeni giren ortakların tescil ve ilanı üçüncü kişilere karşı önem arz ederken; iç ilişkide ise değişiklik, hukuken geçerli olarak meydana geldiğinde hüküm ifade eder. Şirkete yeni ortak alınması ve payın devri suretiyle ortaklık kazanımı başlıklarında değinildiği üzere; şirkete yeni giren kişinin diğer ortaklarla ilişkisi sözleşme serbestisi kapsamında arzu edildiği gibi düzenlenebilir (TTK m.217). Dış ilişkiye yönelik, şirket alacaklarına karşı yükümlü bulunulan borçlar bakımından ise şirkete yeni giren kişi, girme tarihinden evvel doğmuş olsa bile, şirketin borçlarından ve taahhütlerinden diğer ortaklarla birlikte müteselsilen ve bütün malvarlığı ile sorumlu olur. Buna aykırı olarak düzenlenen sözleşme hükümleri ise üçüncü kişiler hakkında geçerli olmayacaktır (TTK m.236). 400 Kollektif Şirkette Ortaklığa Giriş SONUÇ Birbirine güvenen ve birbirlerinin şahsi niteliklerini bilen iki veya daha fazla gerçek kişinin bir araya gelmesiyle oluşan kollektif şirket bu yönüyle tipik bir şahsi şirket kimliği ihtiva etmektedir. Ancak bu yönüyle kollektif şirket, uygulamada şahıslar tarafından pek tercih edilmeyen bir şirket türüdür. 2011 tarihli 6102 sayılı yeni TTK’da kollektif şirketi düzenleyen hükümler, eski kanun olan 6762 sayılı TTK’dan fazlaca bir değişiklik yapılmadan çoğu hüküm korunarak yeni kanuna aktarılmıştır. Kollektif şirkette ortaklığa girmenin hüküm ve sonuçlarına dair hem eski TTK’da hem de yeni TTK’da hüküm bulunmamaktadır. Kollektif şirketlere ilişkin kısımda düzenleme bulunmaması durumlarında, öncelikle TMK’nın tüzel kişilere ilişkin genel hükümlerinin, bu kısımlarda hüküm bulunmayan durumlarda ise TBK’nın adi şirkete dair hükümlerinin uygulanacağı Kanunda ifade edilmiştir (TTK m.126). Kollektif şirkette ortaklığa giriş üç tür yolla mümkündür. Bunlar; şirkete yeni ortak alınması, payın devredilmesi suretiyle ortaklığa giriş ve ortağın ölümü halinde mirasçıların şirkete ortak olması halleridir. 6102 sayılı TTK’da, bu durumlardan yalnızca ortağın ölümü halinde mirasçının şirket ortak olabilmesi hali düzenlenmiştir. Ancak bu durum da ‘‘Ortağın Ayrılması’’ başlığı altında ifade edilmiştir. Dolayısıyla, şirkete yeni ortak alınması ve payın devri suretiyle ortaklık sıfatının kazanımı hususlarında TBK’da yer alan adi şirkete ilişkin hükümler uygulanama alanı bulacaktır. Adi şirket, bir ticaret şirketi olmaması nedeniyle TBK’da düzenlenen ve tüzel kişiliği haiz olmayan bir oluşumdur. Kanun koyucunun, kollektif şirketlere ilişkin ortaklığa giriş durumlarında düzenleme yapmayıp adi şirket hükümlerinin uygulanması yoluna gitmesi, kollektif şirketi tüzel kişiliği haiz bir adi şirket hükmünde gördüğünün kabulüdür. Ancak, gerek sorumluluk, gerek şirket kuruluşu konularında birbirinden hayli farklılık arz eden bu iki şirket türüne ilişkin ayrı ayrı düzenlemeler ihdas edilmesi gerekirken, kanun koyucunun, ‘‘adi şirkete dair hükümlerin niteliklerine uygun düştüğü ölçüde kollektif şirkete de uygulanması’’ hususunu kabul etmesi, kanaatimizce isabetli olmamıştır. Çünkü, adi şirketin tüzel kişiliği haiz bir yapısı bulunmaması nedeniyle, adi şirket hükümlerini kollektif şirketin niteliğine uygun düştüğü ölçüde uygulamak, konuyla ilgili ihtilaflarda yorum yoluna gidilmesini gerekli kılmaktadır. Bu durum da belli konularda tartışmaları beraberinde getirmiştir. Netice itibarıyla, hukuki uyuşmazlıkların daha sağlıklı çözümlenebilmesi adına, TTK’da kollektif şirkete girişe ilişkin düzenlemeler ihdas edilmesi kanaatindeyiz. Yunus Alhan \| 401 Yunus Alhan \| 401 KAYNAKÇA KAYNAKÇA Alışkan M, ‘‘İşletme Konusu ve 6102 Sayılı Türk Ticaret Kanununa Göre Tica- ret Şirketlerinin Ehliyeti ve Temsili’’, MÜHFHAD, 18(2), 2012, Erişim Tarihi: 30 Mart 2022, s. 207- 219. Ayar A, ‘‘Adi Ortaklık Sözleşmesinde İç ve Dış İlişkiler’’, Yeditepe Üniversitesi Sosyal Bilimler Enstitüsü Özel Hukuk Ana Bilim Dalı (Doktora Tezi), 2015, Erişim Tarihi: 25 Mart 2022. Ayhan R/ Çağlar H, Ticari İşletme Hukuku Genel Esaslar, 11. Bası, Yetkin Yayın- ları 2018. Ayhan R/ Çağlar H/ Özdamar M, Şirketler Hukuku Genel Esaslar, 2. Baskı, Yet- kin Yayınları 2020. Bahtiyar M, Ortaklıklar Hukuku, 13. Baskı, Beta Yayınevi 2019. Bozkurt T, Şirketler Hukuku, 9. Baskı, Legem Yayıncılık 2018. Derdiman F, ‘‘Adi Ortaklık Yapısındaki Değişiklikler’’, İstanbul Üniveristesi Sos- yal Bilimler Enstitüsü Özel Hukuku Ana Bilim Dalı (Yüksek Lisans Tezi), 2021, Erişim Tarihi 28 Mart 2022. Öğütcü Ö. F, ‘‘Adi Ortaklıkta İç İlişkiler’’, Hacettepe Üniversitesi Sosyal Bilimler Enstitüsü Özel Hukuk Ana Bilim Dalı (Yüksek Lisans Tezi), 2019, Eri- şim Tarihi: 27 Mart 2022. Poroy R/ Tekinalp Ü/ Çamoğlu E, Ortaklıklar Hukuku - I, 14. Bası, Vedat Ki- tapçılık 2019. Pulaşlı H, Şirketler Hukuku Genel Esaslar, 5. Baskı, Adalet Yayınevi 2017. Şener O. H, Teorik ve Uygulamalı Ortaklıklar Hukuku Ders Kitabı, 4. Bası, Seçkin Yayınevi 2019. Yıldırım A, Türk Borçlar Hukuku Genel Hükümler, 6. Baskı, Adalet Yayınevi 2017. 402 \| Kollektif Şirkette Ortaklığa Giriş
https://openalex.org/W3197496490	https://hal.science/hal-03863241/document	English	null	Ultra-cool dwarfs in wide binaries from Gaia	HAL (Le Centre pour la Communication Scientifique Directe)	2,021	public-domain	3,037	To cite this version: Céline Reylé, Victor Gimenez Sanchez, Nadège Lagarde. Ultra-cool dwarfs in wide binaries from Gaia. The 20.5th Cambridge Workshop on Cool Stars, Mar 2021, Online, France. 10.5281/zenodo.4730624. hal-03863241 2 Sample and method We started from a sample of 16,526 UCDs selected from the precise Gaia DR2 colour absolute magnitude diagram (see Fig. 1), using the data ﬁltering and selection described by Reylé (2018). We query the Gaia DR2 archive1 (Salgado et al. 2018) for companion stars matching the following criteria: ) The Gaia mission Gaia Collaboration et al. (2016) of- fers the means to uncover nearby UCDs through astrometric, rather than purely photometric, selection. On its completion, Gaia should provide high precision astrometry and multi- epoch photometry for over 1000 L- and T-type dwarfs, and po- tentially tens of thousands of late M dwarfs (Sarro et al. 2013). Reylé (2018); Smart et al. (2019); Scholz (2020); Gaia Col- laboration et al. (2020b); and others have undertaken the sys- tematic searches of UCDs in Gaia second (Gaia DR2, Gaia Collaboration et al. 2018) and Early Third (Gaia EDR3, Gaia Collaboration et al. 2020a) data releases. Nearly 14,000 pre- viously non characterized UCDs candidates have been iden- tiﬁed. These are evenly distributed across the sky, ﬁlling in “missing” populations in the Southern hemisphere and Galac- tic plane. (1) ρ < 100ϖ; ∆ϖ < max[1.0, 3σϖ]; ∆µ < 0.1µ; ∆θ < 15◦ (1) Where ρ is the query radius, equivalent to a maximum pro- jected separation of 100,000 AU from the source star (a con- servative upper limit according to Caballero (2009)), ∆ϖ is the parallax range in the LOS direction, ∆µ is the diﬀerence in proper motion and ∆θ is the angular diﬀerence between proper motion vectors. These criteria are discussed in Smart et al. (2019). This search results in 3,445 stars in multiple systems. How- ever the method used is likely to ﬁnd a non-negligible amount of by chance aligned stars. Since no radial velocity measure- ments are available for most of the stars nor can we calcu- late the mass of the system in an accurate enough manner to ensure it is gravitationally bound, we must ﬁnd an alterna- tive way of assessing the probability of a star being a binary. We thus follow a similar method for Bayesian analysis as de- scribed by Hartman & Lépine (2020). In order to compute the bayesian probabilities, we perform a "real" and a "spurious" search. The ﬁrst one is done within a radius of 1 from the original UCD positions at a parallax of ϖ±max[1, 3σϖ]. 1https://gea.esac.esa.int/archive/ 1 Introduction et al. 2020). Improving our knowledge of binary systems in- cluding UCDs can bring constraints on their physics and for- mation mechanisms. Ultra-Cool dwarfs (UCDs) are the lowest-mass, coldest, and faintest products of star formation, making them diﬃ- cult to study. They were deﬁned by Kirkpatrick et al. (1997) as objects with spectral types M7 and later, through L, T, and Y types, have masses ≲0.1 M⊙, and eﬀective temper- atures ≲2700 K. UCDs are of particular interest since they include both very low-mass stars that slowly fuse hydrogen, and brown dwarfs, which have insuﬃcient mass (below about 0.075 M⊙) to sustain hydrogen fusion in their cores, and cool down with time. These objects are nevertheless abundant (> 20% of all stars) and are key probes in Galactic studies. More- over, they are known to host exoplanets, and are ideal targets for searches for potentially habitable terrestrial planets (such as the recent discoveries around TRAPPIST-1, Gillon et al. 2017). In this work we use the sample of UCDs revealed by Gaia and search for companions from their common parallax and proper motion to investigate their binarity. The sample and the method are described in Sect. 2. The results are given in Sect. 3. Sect. 4 gives the conclusions and perspectives. Abstract Ultra-cool dwarfs (UCDs) serve as a link between the lowest mass stars and the brown dwarfs. Gaia DR2 revealed thousands of ultra-cool dwarfs not identiﬁed as such before. In this work, we searched for companions to those in the Gaia catalogue, from their common proper motions and parallaxes. We found about 2,300 stars in multiple systems, about 1,300 of them being UCDs out of the original 16,500 sample. Among the companions, there are several tens of young stellar objects and white dwarfs, but also new M7-L2 candidates. Such a sample can be used to better understand the binary properties of UCDs and to better characterize the physics of UCDs thanks to the characterization of higher mass companions. Ultra-cool dwarfs in wide binaries from Gaia Céline Reylé ,1 Victor Sanchez Gimenez ,1 Nadège Lagarde 1 1 Institut UTINAM, CNRS UMR6213, Univ. Bourgogne Franche-Comté, OSU THETA Franche-Comté-B Besançon, BP 1615, 25010 Besançon Cedex, France HAL Id: hal-03863241 https://hal.science/hal-03863241v1 Submitted on 27 Nov 2023 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. Public Domain The 20.5th Cambridge Workshop on Cool Stars, Stellar Systems, and the Sun Edited by S. J. Wolk, A. K. Dupree, H. M. Günther The 20.5th Cambridge Workshop on Cool Stars, Stellar Systems, and the Sun Edited by S. J. Wolk, A. K. Dupree, H. M. Günther Ultra-cool dwarfs in wide binaries from Gaia Céline Reylé ,1 Victor Sanchez Gimenez ,1 Nadège Lagarde 1 1 Institut UTINAM, CNRS UMR6213, Univ. Bourgogne Franche-Comté, OSU THETA Franche-Comté-Bourgogne, Observatoire de Besançon, BP 1615, 25010 Besançon Cedex, France 2http://simbad.u-strasbg.fr 3.1 Multiplicity frequency Our sample contains 2,332 stars in systems out of which 1,286 are UCDs distributed in 926:182:178 binary, tertiary and higher order systems, respectively. This yields to a mul- tiplicity rate of 7.8% for the UCDs. In comparison, Winters et al. (2019) searches for companions to M0 to M9.5 dwarfs including close pairs and ﬁnds a multiplicity rate of 26.8%. Since our sample is devoid of close binaries and we do not search for companions to the brighter M0 to M6 dwarfs, it is reasonable to expect our sample to exhibit a lower multiplicity rate. 2 Sample and method The Yet they remain elusive due to their low brightness, and the modeling of their cold and complex atmosphere, and in par- ticular the onset of dust formation, remains challenging. Bi- naries with UCDs are expected to be relatively common, with binary systems containing M dwarfs accounting for 26.8% of the M dwarf population in the solar neighbourhood according to (e.g. Winters et al. 2019). Systems with a higher mass and well characterized primary are benchmarks to constrain the atmospheric parameters and ages of the UCDs (e.g. Marocco 1 C. Reylé, V. Sanchez Gimenez, N. Lagarde Figure 1: Colour absolute magnitude diagram from Gaia DR2 of 3,716,407 stars with reliable astrometric and photometric solutions, avoiding Galactic regions with high extinction, and after removing stars showing color excess using G-J (Reylé 2018). The locus of the UCDs is indicated by the blue rectan- gle. It is deﬁned from the expected position based on a sample of spectroscopically characterized UCDs. Figure 2: Distribution of logarithmic projected separation be- fore (grey) and after (blue) ﬁltering binaries with Bayes prob- ability less than 0.8. The ultra wide binaries are dominated by chance aligned pairs. Figure 2: Distribution of logarithmic projected separation be- fore (grey) and after (blue) ﬁltering binaries with Bayes prob- ability less than 0.8. The ultra wide binaries are dominated by chance aligned pairs. Figure 1: Colour absolute magnitude diagram from Gaia DR2 of 3,716,407 stars with reliable astrometric and photometric solutions, avoiding Galactic regions with high extinction, and after removing stars showing color excess using G-J (Reylé 2018). The locus of the UCDs is indicated by the blue rectan- gle. It is deﬁned from the expected position based on a sample of spectroscopically characterized UCDs. initial sample is probably not complete due to the strict ﬁlters applied by Reylé (2018) in order to get robust candidates. 3.2 Projected separations Fig. 2 shows the projected separation distribution, in astro- nomical unit, of the binary systems, before (in grey) and after (in blue) the ﬁltering from the bayesian analysis. second one is done the same way but shifting the search by 4200 arcsec in declination. Proper motion diﬀerence and sep- aration distributions are then computed in both cases and used to deﬁned the diﬀerence Bayes probability of being a true or false binary with proper motion diﬀerence and separation as priors. g y y It can be divided in two groups: wide binaries (< 104 AU, herafter WB), and ultra-wide binaries (> 104 AU, here- after UWB). The distribution for WB follows a similar trend as the one shown by Hartman & Lépine (2020) or Arenou et al. (2018), with increasing binary number as separation de- creases, up to some limit likely due to limiting resolution. In terms of angular separation distribution our sample shows a sharp drop at separation < 6 arcsec. This is higher than the 0.4 arcsec limiting resolution found by Arenou et al. (2018), because our sample is dominated by rather faint stars. The change of distribution in the WB and UWB regimes might indicate a diﬀerent formation mechanism for WB and UWB (see e.g. Tokovinin 2017). p The selection of stars with probability > 0.8 of being binary returns 1,534 stars in systems. Adding all stars in higher order systems, we ﬁnally get 2,332 stars. Acknowledgments This work has made use of data from the European Space Agency (ESA) mission Gaia (https://www.cosmos.esa. int/gaia), processed by the Gaia Data Processing and Anal- ysis Consortium (DPAC, https://www.cosmos.esa.int/ web/gaia/dpac/consortium). Funding for the DPAC has been provided by national institutions, in particular the insti- tutions participating in the Gaia Multilateral Agreement. This work was supported by the "Programme National de Physique Stellaire" (PNPS) of CNRS/INSU co-funded by CEA and CNES. Figure 4: Colour absolute magnitude diagram of the stars in systems. The colour indicates the primary spectral type (white dwarfs are in grey) and the size indicates the secondary spec- tral type. L2, 2 L3), 13 of which were not in the initial UCDs sample, meaning they are companions of M7 and later UCDs. There are seven systems with more than one L dwarf. Interestingly enough, only two of them are binary, the others having mul- tiple M-L stars. However, only three of the L dwarfs among the 13 in these systems have 2MASS (Two Micron All Sky Survey; Skrutskie et al. 2006) counterpart and the others need conﬁrmation. L2, 2 L3), 13 of which were not in the initial UCDs sample, meaning they are companions of M7 and later UCDs. There are seven systems with more than one L dwarf. Interestingly enough, only two of them are binary, the others having mul- tiple M-L stars. However, only three of the L dwarfs among the 13 in these systems have 2MASS (Two Micron All Sky Survey; Skrutskie et al. 2006) counterpart and the others need conﬁrmation. 3.3 Spectral type distribution We used 190,000 objects with spectral type from a SIM- BAD2 (Wenger et al. 2000) query to deﬁne a spectral type cal- ibration from the median absolute magnitude MG, and used it to estimate the spectral types of the companions. The distribu- tion of primaries and secondaries (and higher order multiplic- ity objects) is shown in Fig. 3. The secondaries distribution peaks at M7 due to the selection of our starting UCDs sample. The primaries distribution peak around M3 since there are the most numerous stars in the Galaxy. We also compared this result with the expected UCD bi- narity from the Gaia Universe Model Snapshot (Robin et al. 2012, GUMS), a simulator developped for Gaia, and based on a stellar population synthesis model of the Galaxy. We re- trieved stars within 100 pc with similar positions in the colour absolute magnitude diagram as our initial UCD sample. 6.2% are in wide binary systems (orbital radius larger than 500 AU. In comparison, our sample shows 5.5% of UCDs being wide binaries, which is consistent with the GUMS result since our Fig. 4 shows the systems in the colour absolute magnitude diagram. The lowest spectral type found is a L3 dwarf which might be a wide companion to the already known binary sys- tem LP 395-8 A-B at a projected separation of 42,000AU. The total sample contains 57 L type dwarfs (30 L0, 20 L1, 5 2 Zenodo, 2021 The 20.5th Cambridge Workshop on Cool Stars, Stellar Systems, and the Sun The 20.5th Cambridge Workshop on Cool Stars, Stellar Systems, and the Sun Figure 3: Spectral type distribution of the system components. Out of all the multiple systems, four were found to belong to star forming regions, with at least one Young Stellar Object (hereafter YSO) candidate in each group. They lie on the right part of the main sequence. A group of twelve stars are found in the Barnard 59 cloud in the Pipe Nebula. Large parts of this region are obscured by dust so it is likely that more sources are missing. The next most populated group with YSOs is located in the Lupus I cloud and has eight members in our sample. 4 Conclusion UCDs represent a high percentage of the stars in our neigh- bourhood, yet they are largely undiscovered because of their typically faint magnitudes. Multiple systems with UCDs of- fer additional constraints which make it possible to calculate physical properties of each star from orbital dynamical pa- rameters. Since wide systems with UCDs are expected to ori- gin from diﬀerent formation mechanisms than systems with smaller separations, studying wide and ultra-wide systems can give additional hints on the formation mechanisms of such stars. Figure 3: Spectral type distribution of the system components. Figure 4: Colour absolute magnitude diagram of the stars in systems. The colour indicates the primary spectral type (white dwarfs are in grey) and the size indicates the secondary spec- tral type. Starting from a sample of 16,526 UCDs, we found 1,286 belonging to a multiple system. Most of them were unknown prior to Gaia DR2, and 85 UCDs, not included in our original UCD sample, are found to be companions. These systems are divided into wide and ultra-wide systems according to their projected separations and follow diﬀerent distributions with projected separation which are consistent with expectations from literature. p We will confront our ﬁndings with recent similar searches (Gaia Collaboration et al. 2020b; El-Badry et al. 2021) using Gaia EDR3. We started spectroscopic follow-up of these pairs (see Low et al. 2021) to learn UCDs physical properties from the higher mass primary. References Arenou, F., Luri, X., Babusiaux, C., Fabricius, C., Helmi, A., et al. 2018, A&A, 616, A17. Caballero, J. A. 2009, A&A, 507, 251–259. ISSN 1432-0746. El-Badry, K., Rix, H.-W., & Heintz, T. M. 2021, MNRAS. Gaia Collaboration, Brown, A. G. A., Vallenari, A., Prusti, T., de Bruijne J H J et al 2018 A&A 616 A1 Caballero, J. A. 2009, A&A, 507, 251–259. ISSN 1432-0746. El-Badry, K., Rix, H.-W., & Heintz, T. M. 2021, MNRAS. We also found 48 white dwarf companions to UCDs. 65% of these systems has a separation lower than 104AU, com- pared to 48% for the full sample. This is reasonable since wide and ultra-wide binaries are relatively instable and easily disrupted, old systems are less likely to have survived. White dwarf and UCD binaries are unlikely to have formed as bi- naries (from processes that may result in wide binaries such as core fragmentation), a likely mechanism of formation for these systems is through cluster dissolution (see for instance Kouwenhoven et al. 2010). Gaia Collaboration, Brown, A. G. A., Vallenari, A., Prusti, T., de Bruijne, J. H. J., et al. 2018, A&A, 616, A1. Gaia Collaboration, Brown, A. G. A., Vallenari, A., Prusti, T., de Bruijne, J. H. J., et al. 2020a, arXiv e-prints, arXiv:2012.01533. Gaia Collaboration, Prusti, T., de Bruijne, J. H. J., Brown, A. G. A., Vallenari, A., et al. 2016, A&A, 595, A1. Gaia Collaboration, Smart, R. L., Sarro, L. M., Rybizki, J., Reylé, C., et al. 2020b, arXiv e-prints, arXiv:2012.02061. 3 Zenodo, 2021 C. Reylé, V. Sanchez Gimenez, N. Lagarde Gillon, M., Triaud, A. H. M. J., Demory, B.-O., Jehin, E., Agol, E., et al. 2017, Nature, 542, 456. g Hartman, Z. D. & Lépine, S. 2020, ApJS, 247, 66. Kirkpatrick, J. D., Henry, T. J., & Irwin, M. J. 1997, AJ, 113, 1421. Kouwenhoven, M. B. N., Goodwin, S. P., Parker, R. J., Davies, M. B., Malmberg, D., et al. 2010, MNRAS, 404, 1835. Low, R., Burgasser, A. J., Reylé, C., Gerasimov, R., Hsu, C.- C., et al. 2021, Research Notes of the American Astronom- ical Society, 5, 26. y Marocco, F., Smart, R. L., Mamajek, E. E., Sarro, L. M., Bur- gasser, A. J., et al. 2020, MNRAS, 494, 4891. g Reylé, C. 2018, A&A, 619, L8. Robin, A. C., Luri, X., Reylé, C., Isasi, Y., Grux, E., et al. 2012, A&A, 543, A100. C. Reylé, V. Sanchez Gimenez, N. Lagarde References Salgado, J., González-Núñez, J., Gutiérrez-Sánchez, R., Car- los Segovia, J., Durán, J., et al. 2018, Gaia DR2 documen- tation Chapter 13: Construction of the archive. Gaia DR2 documentation. Sarro, L. M., Berihuete, A., Carrión, C., Barrado, D., Cruz, P., et al. 2013, A&A, 550, A44. Scholz, R. D. 2020, A&A, 637, A45. , , , , Skrutskie, M. F., Cutri, R. M., Stiening, R., Weinberg, M. D., Skrutskie, M. F., Cutri, R. M., Stiening, R., W Schneider, S., et al. 2006, AJ, 131, 1163. Schneider, S., et al. 2006, AJ, 131, 1163. Smart, R. L., Marocco, F., Sarro, L. M., Barrado, D., Beamín, J. C., et al. 2019, MNRAS, 485, 4423. Tokovinin, A. 2017, MNRAS, 468, 3461. Wenger, M., Ochsenbein, F., Egret, D., Dubois, P., Bonnarel, F., et al. 2000, A&AS, 143, 9. Winters, J. G., Henry, T. J., Jao, W.-C., Subasavage, J. P., Chatelain, J. P., et al. 2019, AJ, 157, 216. 4 Zenodo, 2021
https://openalex.org/W2913868037	https://research-information.bris.ac.uk/ws/files/187398050/FINAL_PUBLISHED.pdf	English	null	Investigating the role of elastostatic stress transfer during hydraulic fracturing-induced fault activation	Geophysical journal international	2,019	cc-by	16,054	Kettlety, T., Verdon, J., Werner, M., Kendall, J. M., & Budge, J. (2019). Investigating the role of elastostatic stress transfer during hydraulic fracturing-induced fault activation. Geophysical Journal International, 217(2), 1200-1216. Article ggz080. https://doi.org/10.1093/gji/ggz080 Publisher's PDF, also known as Version of record License (if available): CC BY Link to published version (if available): 10.1093/gji/ggz080 Link to publication record on the Bristol Research Portal PDF-document This is the final published version of the article (version of record). It first appeared online via Oxford University Press at https://academic.oup.com/gji/article/217/2/1200/5315764 . Please refer to any applicable terms of use of the publisher. Kettlety, T., Verdon, J., Werner, M., Kendall, J. M., & Budge, J. (2019). Investigating the role of elastostatic stress transfer during hydraulic fracturing-induced fault activation. Geophysical Journal International, 217(2), 1200-1216. Article ggz080. https://doi.org/10.1093/gji/ggz080 Kettlety, T., Verdon, J., Werner, M., Kendall, J. M., & Budge, J. (2019). Investigating the role of elastostatic stress transfer during hydraulic fracturing-induced fault activation. Geophysical Journal International, 217(2), 1200-1216. Article ggz080. https://doi.org/10.1093/gji/ggz080 SU MMARY We investigate the physical processes that generate seismicity during hydraulic fracturing. Fluid processes (increases in pore pressure and poroelastic stress) are often considered to be the primary drivers. However, some recent studies have suggested that elastic stress interactions may signiﬁcantly contribute to further seismicity. In this work we use a microseismic data set acquired during hydraulic fracturing to calculate elastic stress transfer during a period of fault activation and induced seismicity. We ﬁnd that elastic stress changes may have weakly promoted initial failure, but at later times stress changes generally acted to inhibit further slip. Sources from within tight clusters are found to be the most signiﬁcant contributor to the cumulative elastic stress changes. Given the estimated in situ stress ﬁeld, relatively large increases in pore pressure are required to reach the failure envelope for these faults—on the order of 10 MPa. This threshold is far greater than the reliable cumulative elastic stress changes found in this study, with the vast majority of events receiving no more than 0.1 MPa of positive CFS, further indicating that elastic stress changes were not a signiﬁcant driver, and that interaction with the pressurized ﬂuid was required to initiate failure. Thus, cumulative stress transfer from small events near the injection well does not appear to play a signiﬁcant role in the reactivation of nearby faults. relieved from the small faulting and fracturing that immediately accompanies ﬂuid injection. However, if nearby faults and fractures receive enough elastic Coulomb stress change from the preceding events, bringing them closer to failure, by the time injection is halted due to the trafﬁc light being exceeded, it may already be too late. In tectonic settings the transfer of static stress is widely used in the estimation of seismic hazard, where it is applied to the distribution of aftershock sequences (Stein 1999; Steacy et al. 2004). These Coulomb models have also been applied to ﬂuid related triggering processes such as the seismicity associated with magmatism (Toda et al. 2002). Recent studies (e.g. Catalli et al. 2013; Sumy et al. 2014; Pennington & Chen 2017) have concluded that elastic stress transfer between induced events can also play a signiﬁcant role during a sequence of induced seismicity. Investigating the role of elastostatic stress transfer during hydraulic fracturing-induced fault activation ety, James P. Verdon, Maximilian J. Werner, J. Michael Kendall and Tom Kettlety, James P. Verdon, Maximilian J. Werner, J. Michael Kendall and Jessica Budge Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 g School of Earth Sciences, University of Bristol, Wills Memorial Building, Queen’s Road, Bristol BS8 1RJ, UK. E-mail: tom.kettlety@bristol.ac.uk School of Earth Sciences, University of Bristol, Wills Memorial Building, Queen’s Road, Bristol BS8 1RJ, UK. E-ma Accepted 2019 February 7. Received 2019 February 4; in original form 2018 July 30 University of Bristol – Bristol Research Portal General rights This document is made available in accordance with publisher policies. Please cite only the published version using the reference above. Full terms of use are available: http://www.bristol.ac.uk/red/research-policy/pure/user-guides/brp-terms/ doi: 10.1093/gji/ggz080 Geophys. J. Int. (2019) 217, 1200–1216 doi: 10.1093/gji/ggz080 Advance Access publication 2019 February 11 doi: 10.1093/gji/ggz080 Geophys. J. Int. (2019) 217, 1200–1216 Advance Access publication 2019 February 11 GJI Seismology C⃝The Author(s) 2019. Published by Oxford University Press on behalf of The Royal Astronomical Society. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. 1 I N T RO D U C T I O N Hydraulic fracturing (HF; Clarke et al. 2014; Bao & Eaton 2016), waste water injection (Keranen et al. 2013), carbon capture and stor- age (CCS; Verdon et al. 2013a) and the enhancement of geothermal systems (EGS; Schoenball et al. 2012) have all shown the abil- ity to stimulate seismicity. Numerous felt earthquakes have been linked to injection across North America and Europe, with magni- tudes reaching as high as MW 5.8 (Keranen & Weingarten 2018). Increases in pore pressure and poroelastic stresses are considered to be the dominant physical mechanism responsible in areas of an- thropogenic seismicity, reducing the effective normal stresses acting on nearby faults and fractures and thus promoting failure (Raleigh et al. 1976). However, the extent to which faulting and fracturing near the well can promote more distant seismicity is not well es- tablished. The uncertainty surrounding the continued propagation of seismicity once initial faulting has occurred has signiﬁcance for the mitigation strategies currently being used in certain jurisdic- tions. ‘Trafﬁc-light’ schemes function under the assumption that if a smaller event (e.g. M 0.5 in the UK) is detected during injec- tion, halting further operations can act to mitigate a larger, possibly damaging, earthquake (Green et al. 2012; Clarke et al. 2014). This assumption hinges on a transient change to the stress state produced from the ﬂuid permeating the rock mass, as well as stress being Displacement on a fault changes the surrounding stress ﬁeld. This stress change will act on nearby faults and fractures, and can promote or inhibit further slip. The Coulomb failure stress change, CFS, acting on a fault is an effective tool for examining the stress accumulated during a sequence of events: C FS = τ + μ′σn, (1) (1) C FS = τ + μ′σn, with change in shear stress τ and change in normal stress σ n (positive extensive) resolved onto the fault plane, and effective co- C⃝The Author(s) 2019. Published by Oxford University Press on behalf of The Royal Astronomical Society. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. 1200 Stress transfer during hydraulic fracturing 1201 in the triggering of speciﬁc event clusters after shut-in occurred. Maghsoudi et al. 1 I N T RO D U C T I O N (2013) cal- culated CI in excess of 75 per cent for induced events associated with the Basel EGS, as well as giving a treatment of uncertain- ties in focal mechanism (FM), location, and frictional parameters. During the 2011 Prague, OK sequence, Sumy et al. (2014) found that the MW 5.0 foreshock, which is considered to have been in- duced by the nearby waste water injection, promoted the MW 5.7 main shock by elastic Coulomb stress transfer. Pennington & Chen (2017) also studied the events associated with waste water injection in Oklahoma, during the 2016 Pawnee sequence. They found that the three foreshocks promoted failure of the MW 5.8 main shock, which went on to promote failure on 67 per cent of the events that occurred after it. Coulomb stress changes resulting from microseis- mic events recorded during HF were calculated by Vasudevan & Eaton (2011), showing that non-double-couple source mechanisms have a signiﬁcant effect on the resulting stress maps. Schoenball et al. (2012) investigated the static Coulomb stress changes for the seismicity associated with an EGS, neglecting any pore pressure change. They found that elastic Coulomb stresses may have only played a small role in promoting failure for the majority of the seismicity associated with the injection, but were more signiﬁcant The variability of induced seismicity between different sites and regions (e.g. Verdon et al. 2016) has been linked to differences in the background stress ﬁeld (King et al. 2014; G¨obel 2015). Fault orientation has been shown to be connected to the occurrence of induced seismicity, with the MW > 5 events associated with waste- water injection in Oklahoma occurring on planes well oriented to the background stresses (Keranen et al. 2013; Alt & Zoback 2017). Thus, the calculated elastic stress changes need to be considered in the context of the in situ stress ﬁeld acting on the individual fracture or fault planes. The conditional probability of failure given the in situ stress ﬁeld can be determined through a quantitative risk assessment (QRA) methodology developed in Chiaramonte et al. (2007) and Walsh & Zoback (2016). QRA methods have been applied to various geomechanical issues, such as wellbore stability (Moos et al. 2003) and CCS caprock integrity (Chae & Lee 2015). Following the approach of Walsh & Zoback (2016), the uncertainties are included in the calculation of the required pore pressure change to initiate failure (given the Mohr-Coulomb slip criterion). 1 I N T RO D U C T I O N Microseismic monitoring during operations has also repeatedly indicated the presence of smaller scale injection-induced fault activation (BC Oil and Gas Commission 2012). The elastic Coulomb stress changes acting on a plane due to any previous slip on surrounding faults can then be calculated using the Okada equations (Okada 1992), identifying if slip was encouraged or discouraged by the preceding fault movements. The overall effect of stress changes acting on an earthquake population is measured using the Coulomb index (CI)–the percentage of events with positive CFS (Hardebeck et al. 1998). A CI signiﬁcantly above 50 per cent is consistent with a population where elastic stress changes actively promoted slip. We use the PSCMP code of Wang et al. (2006) to model the cu- mulative elastic stress changes during a period of HF-induced fault reactivation. We use the full moment tensor solutions, including the non-double couple fault motion. Sensitivity analyses are then conducted to constrain the effects of focal mechanism uncertainty on CFS through Monte Carlo methods, as developed in Catalli et al. (2013) and Meier et al. (2014). We examine the effect of the ﬂuids on the resulting CFS by varying Skempton’s ratio β. Due to the observed tight clustering of events, we also compare the elas- tic stress changes within clusters to the total cumulative CFS to investigate the signiﬁcance of intracluster triggering. This appears to be a relatively simple procedure, but determining the signiﬁcance of a positive CFS signal for a large number of events is non-trivial. One should appropriately consider the uncer- tainties in fault orientation, material properties and slip behaviour. A study by Meier et al. (2014) highlighted the importance of a rigorous statistical treatment of static stress modelling by conducting a thor- ough analysis of the stress changes associated with the 1992 Landers earthquake sequence, assessing the signiﬁcance of positive stress changes with regard to the modelling assumptions and the uncertain- ties in the input parameters. They explored techniques to determine signiﬁcance and the sensitivity of elastic stress models, especially to focal mechanism uncertainty. Further studies have highlighted the effect of elastic stress transfer in some injection-induced earth- quake sequences, with relatively high magnitude events correlated with positive Coulomb stress changes. Catalli et al. 1 I N T RO D U C T I O N (2018) used statistical techniques to show that spa- tiotemporal clusters of events, associated with an HF stimulation in the Horn River Basin (HRB), exhibited evidence for interevent trig- gering. They showed that the occurrence of events deviated from a purely Poisson process, and temporal clustering was present. In their study, while the microseismic event clusters exhibit this aftershock sequence-like pattern, the events deviate from the typical main- shock–aftershock behaviour observed in tectonic sequences. Large numbers of events also exhibit swarm-like behaviour, where the trig- gered events have very similar magnitudes to those that triggered them and, within a given cluster, the largest event is not typically the ﬁrst. efﬁcient of friction μ ′ (Harris 1998). A positive CFS means the stress conditions have changed in such a way as to promote slip, whereas negative acts to inhibit slip. Here, μ′ = μ(1 −β), (2) (2) μ′ = μ(1 −β), where μ′ is used to summarize the complex assumptions relating pore pressure change, the frictional properties of the fault and the effective normal stress. Skempton’s ratio β (Skempton 1954) is used to relate the change in pore pressure change to the change in the applied stress: P = −βσ kk/3. Assuming that the fault zone is homogeneous and isotropic, one can relate the diagonal components of the stress tensor σ kk to the normal stress σ n (Rice 1992; Simpson & Reasenberg 1994). This gives σ kk/3 = σ n, and eqs (1) and (2) are satisﬁed (Harris 1998). This β parameter can be thought of as simulating the frictional stability of a fault in a porous medium. With β > 0, pore pressure changes act against the compressive stress, reducing the effective friction on the fault. In our study we use a microseismic data set acquired during an HF operation in the HRB. The HRB is one of the largest natural gas plays in British Columbia, Canada, and is part of the West- ern Canadian Sedimentary Basin (WCSB). Most of the wells in the HRB are horizontal, multiwell operations deployed from single pads, employing multistage HF to stimulate production from three Devonian shale layers: the Muskwa, Otter Park and Evie formations (Barker 2014). In recent years, multiple felt earthquakes associated with HF have been recorded in the WCSB (Schultz et al. 2015; Bao & Eaton 2016), with magnitudes reaching as high as MW 4.6 (Kao et al. 2018). 2.1 Identifying fault reactivation To determine the minimum magnitude of completeness (MMIN) for each cluster, we use a Kolmogorov-Smirnov test, with MMIN being the smallest magnitude at which the observed popula- tion can be ﬁt with an exponential distribution with a conﬁdence level of 95 per cent. Moment tensors were determined by inversion of polarity data and P- to S-wave amplitudes for an estimated radiation pattern (as in Vavryˇcuk 2014, 2015b). The fault plane and auxiliary planes were discriminated using the in situ stresses, ﬁnding the preferentially aligned nodal plane (NP) in the local stress ﬁeld. This can be used as an estimate of the fault plane, over the auxiliary plane. However, the choice of nodal plane may not be signiﬁcant for this elastic mod- elling, as discussed in Section 3.1. Throughout this work, we refer to a nominal ‘primary’, preferred NP chosen during the inversion, and a ‘secondary’ nodal plane. The resulting magnitude-frequency distributions are shown in Fig. 2. It is immediately apparent that the distribution of b-values is bimodal. The majority of event clusters have high b-values (b > 1.5), implying that they are associated with normal hydraulic fracture propagation, while a smaller number of clusters have b ≈ 1.0, a value similar to tectonic earthquake sequences, indicating that they are associated with fault reactivation (Verdon et al. 2013b; Verdon 2013; Eaton et al. 2014; Eaton & Maghsoudi 2015). We therefore use b < 1.5 as our criteria for identifying microseismic event clusters that represent fault reactivation. Some of the determined MTs include a volumetric component. The vast majority (90 per cent) of events are effectively double- couple (DC) sources (with tensile angles less than 3◦), though some events, especially smaller events closer to the well, have larger non-DC components. Events with signiﬁcant tensile angles (α > 10◦), most likely associated with the opening of fractures though hydraulic stimulation, are generally far smaller in magnitude than those associated with the fault reactivation, with 84 per cent having magnitudes less than MW −0.5. Thus, while the non-DC component of slip is included in the analysis, it will have less of an effect on the overall pattern of stress change than the larger, DC sources. These clusters occurred mostly during two periods of injection, as can be seen in Fig. 3. These are denoted by reactivation periods 1 and 2 (or, RP1 and RP2) and are separated by around 42 hours. 2.1 Identifying fault reactivation Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Initial examination of the microseismic event hypocentres indicated the re-activation of pre-existing faults, with clusters of events map- ping out planar features that extend approximately 400 m below the wells, into the underlying Keg River limestone. The largest events were found in these clusters, with 27 events having magni- tudes greater than MW = 0.0, whereas the overwhelming majority of events, those associated with hydraulic fracture propagation, have magnitudes around −2 < MW < −1. The largest events, and the most obvious downward growth of microseismicity, were seen in proximity to Wells 1 to 4, associated with the stimulation stages in the central portions of these wells (see Fig. 1). Because these events represent the clearest example of fault reactivation within this data set, we focus our analysis on the events associated with these wells and stages. p p y p Event hypocentres were ﬁrst calculated by inverting P- and S- wave traveltimes through a layered anisotropic velocity model. This model was determined from sonic logs, and further calibrated using the perforation shots. Once preliminary locations were found, the velocity model was then linearly tapered at the transitions between the shale layers to more accurately reﬂect the smooth transitions observed in the sonic logs. Events were then relocated using this reﬁned velocity model. Double difference corrections were then ap- plied, reﬁning the relative locations of the events. Seismic moment, source radius and slip were found for each event by ﬁtting a source model to the event displacement spectra (see Stork et al. 2014). The source radii for the microseismic events used in this study are distributed around 15 m, with the majority (96 per cent) under 20 m. A small number (36 of the 923) of higher magnitude events have source radii up to 110 m. To further discriminate fault reactivation from the operationally induced microseismicity associated with fracturing, we examined variations in the b-value of the event magnitude-frequency distri- bution (Gutenberg & Richter 1944). For each stage, we clustered events based on their spatial positions using a density-based clus- tering algorithm DBSCAN (Ester et al. 1996). For each cluster we computed the b-value using the maximum likelihood approach (Aki 1965). T. Kettlety et al. T. Kettlety et al. method is discussed in Section 3.1, and more in Section 4. The uncertainty in the fault geometries, as inferred from the MTs, is the subject of the sensitivity analysis conducted later in this work (Section 3.3). Magnitudes, and thus source radii and slip, are con- sidered to be well constrained, given the standard method by which they were calculated, and the low noise for these events detected on approximately 80 stations in deep borehole arrays. 2 DATA We use a microseismic data set that was acquired during a multi- well, multistage hydraulic fracture treatment conducted in the HRB (described in Baird et al. 2017; Verdon & Budge 2018). 237 stages were completed in the 10 wells drilled, using a toe-heel, zipper-frac injection pattern. Continuous monitoring was provided by three downhole microseismic arrays. For the 119 stages for which we have data, around 92 700 events were recorded, processed and cata- logued. These stages of injection constitute the 10 to 12 most prox- imate stages to the monitoring arrays for each well, and thus are expected to give the best quality data (shown in Fig. 1). Magnitudes (ranging from −2.4 < MW < 0.5), fault radii, stress drops, slips and locations were determined for all events in the catalogue, and around 35 per cent have full moment tensor solutions (MTs). The processing was conducted by a service provider, ESG Solutions. The data is proprietary and is not available for public release. 1 I N T RO D U C T I O N A ‘probability’ of failure for a given P can be determined by a Monte Carlo method of randomly sampling from distributions of the input parameters. This fault slip potential (FSP) can then be used as a proxy for the likelihood of failure for a given ﬂuid pressure increase. l d d b i i h i d l i We conclude our study by interpreting the estimated elastic stress changes in the context of the estimated FSP for the activated faults. 1202 2.1 Identifying fault reactivation Little seismicity occurred in the vicinity of these clusters except during these two periods. The events are shown in Fig. 3 with the same colour as the well with which they are associated. Each reactivation period is associated with an injection stage in Well 2, with fault reactivation clusters located beneath these injection stages. Seismicity continued in these clusters after the Well 2 stages had stopped, at which point injection switched to Well 4. However, the injection stages from Well 4 are over 500 m NE from Well 2, and there is clear lateral separation between the Well 4 events and the Well 2 fault reactivation, with the Well 4 events all appearing to show operationally induced HF events. We conclude that the fault reactivation events are associated solely with the Well 2 stages, and No uncertainties were provided by the operating contractor. Thus, we estimate uncertainties throughout, informed by the methods through which the parameters were determined. As the location determination went through multiple stages of reﬁnement, the ab- solute location uncertainty would be approximately ±50 m, while the relative error may be as low as ±10 m. We go on to estimate the location uncertainty further, by quantifying how diffuse each of the clusters are that map out the presumed fault planes (Fig. 4b). The combined effect of a low relative location error and the modelling Stress transfer during hydraulic fracturing 1203 Figure 1. Hypocentres in plane view (a) and cross-section (b) of all events detected and located during hydraulic stimulation of the 10 wells at the study site. Events are coloured by moment magnitude MW. Wells 1 to 4 are shown as coloured lines, with the other wells being shown in black. The events associated with these wells show the clearest evidence for fault reactivation and are the subject of this study. The red dashed boxes delineate the approximate positions of these hypothesized fault-related events. Also shown in (b) is the 1-D geologic model, showing the depth of the overlying Fort Simpson formation, the three targeted shale layers (Muskwa, Otter Park and Evie), and the underlying Keg River limestone. The wells and corresponding targeted shale layers have matching colours. not those of Well 4. 2.1 Identifying fault reactivation Their apparent contemporaneity with Well 4 injection simply represents the fact that seismicity continued after injection had ceased in Well 2 as is commonly observed during An additional cluster of low b-value seismicity occurred during a later injection stage in Well 2, to the NW of the clusters shown in Fig 3 forming another cluster extending downwards from the Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Stress transfer during hydraulic fracturing 1203 Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 217/2/1200/5315764 by University of Bristol Library user on 14 Figure 1. Hypocentres in plane view (a) and cross-section (b) of all events detected and located during hydraulic stimulation of the 10 wells at the study site. Events are coloured by moment magnitude MW. Wells 1 to 4 are shown as coloured lines, with the other wells being shown in black. The events associated with these wells show the clearest evidence for fault reactivation and are the subject of this study. The red dashed boxes delineate the approximate positions of these hypothesized fault-related events. Also shown in (b) is the 1-D geologic model, showing the depth of the overlying Fort Simpson formation, the three targeted shale layers (Muskwa, Otter Park and Evie), and the underlying Keg River limestone. The wells and corresponding targeted shale layers have matching colours. An additional cluster of low b-value seismicity occurred during a later injection stage in Well 2, to the NW of the clusters shown in Fig. 3, forming another cluster extending downwards from the well. This appears to connect to the diffuse cluster visible at around not those of Well 4. Their apparent contemporaneity with Well 4 injection simply represents the fact that seismicity continued after injection had ceased in Well 2, as is commonly observed during injection-induced fault reactivation (Deichmann & Giardini 2009). 1204 T. Kettlety et al. Figure 2. Frequency–magnitude distributions for event clusters associated with fracturing stages in Wells 1 to 4. Each line corresponds to one cluster. The resulting b-values are clearly bimodal, with most clusters having b > 1.5 (coloured blue/green), and a smaller number of clusters having b < 1.5 (coloured yellow/red). The four low b-value clusters are those shown in Fig. 3, which make up the activity in reactivation periods 1 and 2. Figure 2. 2.1 Identifying fault reactivation Frequency–magnitude distributions for event clusters associated with fracturing stages in Wells 1 to 4. Each line corresponds to one cluster. The resulting b-values are clearly bimodal, with most clusters having b > 1.5 (coloured blue/green), and a smaller number of clusters having b < 1.5 (coloured yellow/red). The four low b-value clusters are those shown in Fig. 3, which make up the activity in reactivation periods 1 and 2. during RP2, although some RP1 events also took place on F2. These planes can be seen projected onto their associated events in Fig. 4. 2200 m depth in Fig. 3. Considering all of the events that were located in this area, two main structures become apparent: a planar feature, and a more diffuse cluster, both extending between 1900 and 2400 m depth. Fig. 4 shows the locations of the low b-value, fault related events, showing that these events map out two pre-existing fault structures both of which strike NE-SW, and dip towards each other. If events are thought to have occurred purely on these conjugate planes, the spread of events around the faults may provide an es- timate for location uncertainty. This estimate would be ±25 m for F1, and ±46 m for F2, as shown by Fig. 4b. However, given the estimated rupture dimensions (∼20 m), it is uncertain as to whether the events observed are actually slip on pre-existing faults, or rather smaller fractures failing in a zone around these structures. Despite their size, all of the largest events (MW > 0) in the catalogue took place along these underlying planar features, we conclude that this region must be fundamentally different from that of a diffuse cloud of fractures, such as those near the wells. To compare these two interpretations, the following analysis will consider both the indi- vidual microseismic event geometries (Section 3.1), as well as the fault planes themselves (Section 3.2). This will also help to con- strain the effect of the uncertainty in the geometries determined from the event moment tensors. The effect of Coulomb stress on the fault planes and the microseismic events should be similar if the event geometries reﬂect that of the larger structures. The occurrence of events appears to be correlated tightly with the injection. We do not observe any clear aftershock-type sequences in either RP, where one large event triggers subsequent smaller ones in a spatiotemporal cluster. 2.1 Identifying fault reactivation This type of behaviour would be indicative of a sequence with signiﬁcant event-event triggering. Event clusters appear more swarm-like, where similar sized events are happening throughout the periods of activity. This swarm-like behaviour is contrasted with the results found by previous studies (Langenbruch et al. 2011; Maghsoudi et al. 2018), where there were clear patterns of triggered temporal clusters during injection-induced seismicity. There is also no clear sign of the characteristic r ∝√t triggering wave front emanating from the point of injection, as exploited in many previous studies to estimate the diffusivity of the medium (Shapiro et al. 1997; Goertz-Allmann et al. 2011). This would be indicative of events triggered by the expanding wave front of pore pressure that results during injection. The elastic Coulomb mod- elling that follows will attempt to probe the ambiguous mechanism behind this sequence of events. 2.2 Stress model for Horn River Basin Assuming that the vertical stress is a principal stress, which is usually the case in sedimentary basins unless signiﬁcant deforma- tion or stress rotation is present, an in situ stress model is deﬁned by the orientation and magnitudes of the principal stresses—the vertical stress SV, the maximum horizontal stress SHmax, and the minimum horizontal stress SHmin—as well as the pore pressure P, and the SHmax orientation φH. Borehole breakouts, density log mea- surements, leak-off tests, shut-in pressures, mud-weights, and the We use a least squares minimization algorithm to ﬁnd the planes that best ﬁt the seismicity. The RP1 plane has a strike φ of 250±10◦ and a dip δ of 80±5◦. The RP2 plane has a φ of 60±15◦and a δ of 75±10◦. Uncertainties for these geometries are estimated given the spread of the events. These two suspected fault structures are denoted fault 1 and 2 (F1 and F2) respectively, named for the period of seismicity in which they were activated: events on F1 were seen mostly during RP1, whereas events of F2 occurred primarily Stress transfer during hydraulic fracturing 1205 Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by Uni Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 3. (a) Hypocentre locations of the studied seismicity, grouped into ‘reactivation periods’ 1 and 2 (RP1 and RP2). Perforation shots are shown as squares, coloured in the same manner as their associated RP. Circles show nearby events considered not to be associated with the fault activation, and are coloured by their associated injection well. The map view of the seismicity is inset. (b) Temporal distribution of the seismicity. Injection durations are shown above, with colours denoting the well from which injection was occurring. Figure 3. (a) Hypocentre locations of the studied seismicity, grouped into ‘reactivation periods’ 1 and 2 (RP1 and RP2). Perforation shots are shown as squares, coloured in the same manner as their associated RP. Circles show nearby events considered not to be associated with the fault activation, and are coloured by their associated injection well. The map view of the seismicity is inset. (b) Temporal distribution of the seismicity. Injection durations are shown above, with colours denoting the well from which injection was occurring. modelling of breakout rotations can be used to constrain estimates in these parameters (Haimson & Cornet 2003; Zoback 2010). 2.2 Stress model for Horn River Basin Light blue downward triangles show fault 1 events, while dark blue upward triangles show fault 2 events. (b) Histograms of lateral distance away from plane for events associated with each fault. Colours are the same as those used in (a). Red lines show ﬁtted normal distributions, where for F1 μ = 0 ±2m and σ = 25 ±2m, and for F2 μ = 0 ±3 and σ = 46 ±2m. These uncertainties represent the 95 per cent conﬁdence intervals. Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 20 Figure 4. (a) Edge on view of events hypocentres used to deﬁne the visible fault planes. Light blue downward triangles show fault 1 events, while dark blue upward triangles show fault 2 events. (b) Histograms of lateral distance away from plane for events associated with each fault. Colours are the same as those used in (a). Red lines show ﬁtted normal distributions, where for F1 μ = 0 ±2m and σ = 25 ±2m, and for F2 μ = 0 ±3 and σ = 46 ±2m. These uncertainties represent the 95 per cent conﬁdence intervals. Hooke’s law for a linear elastic medium. This stress tensor is then resolved onto a receiver plane and rake, and the resulting Coulomb stress change CFS is calculated (eq. 1). All events during RP1 and RP2 are deﬁned as ‘receivers’, and each event that occurred prior to each receiver is treated as ‘source’ of deformation. This gave a total of 1119 receivers (of which 923 have MTs), and potentially 6602 sources (of which 1876 have MTs). Proceeding iteratively through each receiver with a MT, the cumulative elastic stress change is calculated at that receiver’s hypocentre. distance over which the ﬂuid pressure is applied, and reducing the magnitude of the resulting P. Constraining these exact parame- ters requires complex hydromechanical modelling, and is beyond the scope of this work. A least squares ﬁt is applied to ﬁnd the gradients of SHmax, SHmin, SV, and P, and their respective uncertainties (York et al. 2004). The stresses acting at the depth of the reservoir are then found by extrapolation. This gave an SHmax of 77±12 MPa, SHmin of 51±6 MPa, SV of 66±5 MPa, and P of 27±7 MPa. These values broadly agree with those found by Chou et al. 2.2 Stress model for Horn River Basin that, for two other reservoirs in the WCSB, regions of overpressure were common, and correlated with an increased occurrence rate of earthquakes. We were not provided with drilling data for the ﬁeld, and thus do not have a measure of the in situ pore pressure for the shales. We thus use hydrostatic pore pressure as a mean value, and give it an appropriate uncertainty to account for the possibility for overpressure. Uncertainties are also estimated for the other princi- pal stresses (see Walsh & Zoback 2016). These will then be used in the Monte Carlo FSP analysis. Previous studies have estimated the stress regime in the HRB for particular operations (Snelling et al. 2013; Sayers et al. 2016), as well as the frictional properties of the shale and underlying lime- stone (Chou et al. 2011; Hurd & Zoback 2012). There are a number of at-depth stress measurements in the region, which arise from the large number of shale gas operations in the HRB, as well as the adjacent Laird Basin and Montney shale play. One factor not directly accounted for is the increase in pore pres- sure that accompanies HF. The magnitude of the P change, and its spatial extent, is highly dependent on the permeability structure of the reservoir Brown & Ge (2018a). More importantly, it is very sensitive the fracture permeability and extent. If matrix permeabil- ity is very low (∼10 nd; Chalmers et al. 2012; Dong et al. 2017), and fracture permeability is ignored, the P perturbation will be in far more than 10 MPa over a scale of tens of metres (Brown & Ge 2018b). Fractures act to increase bulk permeability, increasing the The stress model used in this study is calculated using data from the World Stress Map (Heidbach et al. 2016). At-depth stress measurements are taken from operator reports catalogued by Bell (2015), with each well located less than 30 km laterally from the site considered here. These measurements are shown in Fig. 5. These data provide a φH of 55±10◦. The pore pressures given appear hy- drostatic. This may be the case only for the formations from which the measurements were taken, or could be the product of a modelling assumption on behalf of the operator. Eaton & Schultz (2018) found 1206 T. Kettlety et al. Figure 4. (a) Edge on view of events hypocentres used to deﬁne the visible fault planes. 2.2 Stress model for Horn River Basin (2011), who constructed a stress model for the HRB shales using borehole observations provided by industry, and Sayers et al. (2016), who used anisotropic seismic attributes to estimate stress conditions. In the Anderson classiﬁcation scheme this is a strike-slip regime, with SHmax > SV > SHmin. Within the uncertainties, the regime could be considered normal faulting, however, that is taken into account in the later Monte-Carlo FSP analysis. The elastic parameters (μ = 0.7 and β = 0.4) used are based on measurements made by (Chou et al. 2011) in the HRB, as well as values obtained for formations of similar type, depth, pressures and porosities (Kohli & Zoback 2013). Sources are modelled as square patches, with dimensions calculated to match the rupture area given by the source spectra. The average patch length is approximately 25 m, with the largest around 190 m. Slip is resolved into the strike and dip components of the plane using the rake and tensile angles (see Vavryˇcuk 2011). Sources are excluded with source- receiver distances less than one source length because CFS values computed using a uniform slip model become unreliable in the very near-ﬁeld (see Meier et al. 2014). This step will have a signiﬁcant effect on the resulting CFS magnitudes and polarities, but will also aid in removing artefacts which result from uncertainties in the slip behaviour and source mechanism (Steacy et al. 2004), and stress drop (Schoenball et al. 2012). The ramiﬁcations of this are discussed in detail in Section 4.1. 3.1 Elastic stress modelling This is in contrast to RP2, where the CI for the entire population is 38 per cent and the windowed values do not exceed 45 per cent. The majority of the larger magnitude events (MW > 0) received positive CFS. However, there is no correlation between the event magnitude and CFS magnitude. For both reactivation periods, no CFS received from the microseismicity exceeds ±1 MPa. There are a number of events (36 per cent) however that received positive CFS greater than the commonly used triggering threshold of 10 kPa (see King et al. 1994; Stein 1999). 60 per cent, and around 12:30 and 14:30 (on the ﬁrst day visible in Fig. 3b) the windowed CI exceeds 70 per cent. This is in contrast to RP2, where the CI for the entire population is 38 per cent and the windowed values do not exceed 45 per cent. The majority of the larger magnitude events (MW > 0) received positive CFS. However, there is no correlation between the event magnitude and CFS magnitude. For both reactivation periods, no CFS received from the microseismicity exceeds ±1 MPa. There are a number of events (36 per cent) however that received positive CFS greater than the commonly used triggering threshold of 10 kPa (see King et al. 1994; Stein 1999). very similar temporal behaviour as for the preferred NP, with peaks around 75 per cent early in RP1, and CI not exceeding 45 per cent in RP2. The similarity should not be surprising however due to the inherent symmetry in the stress change pattern when using a uniform slip model (Meier et al. 2014). This results in generally similar stress change values. The NP ambiguity is further explored in Section 3.3. very similar temporal behaviour as for the preferred NP, with peaks around 75 per cent early in RP1, and CI not exceeding 45 per cent in RP2. The similarity should not be surprising however due to the inherent symmetry in the stress change pattern when using a uniform slip model (Meier et al. 2014). This results in generally similar stress change values. The NP ambiguity is further explored in Section 3.3. 3.1 Elastic stress modelling Using the estimated slips, rupture lengths, and fault plane orienta- tions, the deformation resulting from each event can be modelled. The code PSCMP by Wang et al. (2006) is used to calculate the displacement ﬁeld u due to slip on a square patch. This uses the an- alytical Okada solution of the Green’s functions for a homogeneous elastic half-space (Okada 1992). The model then derives the stain tensor ϵij from u, which is then related to the stress sensor σ ij using Fig. 6 shows resulting CFS values found for RP1 and RP2 when the preferred nodal plane, as identiﬁed during the MT inversion, is used. The Coulomb index for all of the events in RP1 is just below Stress transfer during hydraulic fracturing 1207 Stress transfer during hydraulic fracturing 1207 Figure 5. Stress gradients produced from the Bell (2015) data for the Laird and Horn River basins. The error bars show the uncertainties used for each of the stress parameters (see Walsh & Zoback 2016). Estimates for the stresses acting at the depth of the reservoir are shown in grey boxes. p p gj y f g y f g gure 5. Stress gradients produced from the Bell (2015) data for the Laird and Horn River basins. The error bars show the uncertainties used for each of the ress parameters (see Walsh & Zoback 2016). Estimates for the stresses acting at the depth of the reservoir are shown in grey boxes. Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by Un Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 5. Stress gradients produced from the Bell (2015) data for the Laird and Horn River basins. The error bars show the stress parameters (see Walsh & Zoback 2016). Estimates for the stresses acting at the depth of the reservoir are shown in gre Figure 5. Stress gradients produced from the Bell (2015) data for the Laird and Horn River basins. The error bars show the uncertainties used for each of the stress parameters (see Walsh & Zoback 2016). Estimates for the stresses acting at the depth of the reservoir are shown in grey boxes. 60 per cent, and around 12:30 and 14:30 (on the ﬁrst day visible in Fig. 3b) the windowed CI exceeds 70 per cent. 3.3 Sensitivity analysis We use a Monte Carlo approach to estimate the sensitivity of CFS to focal mechanism uncertainties, as in Catalli et al. (2013) and Meier et al. (2014). Using the ﬁrst 500 events with MTs that oc- curred during the ﬁrst reactivation period, 3000 test catalogues are produced. Each catalogue contains the same 500 events, but with the geometries permuted in three different fashions, giving 1000 catalogues for each type. In the ﬁrst set of catalogues, the NP is randomly selected from the original FM in the catalogue. In the second case, Von Mises distributed rotations are applied to the preferred NP, representing uncertainties in the focal mechanism in- version. As no uncertainties were provided from the processing, a single FM uncertainty of σ = 30◦is given, representing a rea- sonable, perhaps pessimistic, value for this form of MT inversion using borehole-acquired microseismic data (Vavryˇcuk 2014). A ﬁ- nal set of test catalogues are generated with entirely random strikes, dips and rakes (Kagan 2005). This allows us to compare the per- turbed FMs stress changes to an extreme uncertainty scenario, as well as a potential null hypothesis case, where there is no pref- erential alignment of failure planes. The CFS modelling is then conducted in the same manner as in Section 3.1 for each catalogue. The CI of each of the catalogues is then calculated, and the distribu- tions in CI can be compared for each of the three FM perturbation methods. The results of this analysis show a similar pattern to the elastic stress changes as calculated in Section 3.1. In RP1, both the sum and the average stresses resolved across the plane are positive, and the effective CI (the proportion of sample points on the plane with CFS > 0) reaches as high as 78 per cent around the middle of RP1 (Fig. 7). This suggests that the elastic stress changes modestly promoted failure on F1. Similar temporal variations in CI are ob- served as for Section 3.1 for the individual events. However, there is a short period during RP1 where the opposite signal is seen. At around 13: 00, when stresses are resolved across F1, the effective CI acting is at its lowest value: ∼35 per cent. But when the individ- ual events are considered, this early part of RP1 however had the relatively high CI of ∼70 per cent. 3.2 Resolving stresses onto fault structures In addition to computing the Coulomb stress changes for each event, we also evaluate the stress interactions between the microseismic events and the two fault planes (see Section 2.1 for description), as in the latter section of Schoenball et al. (2012). The same ge- omechanical parameters as in Section 3.1 are used, along with the preferred NP geometries. As events on F1 only occurred during the ﬁrst reactivation period, only stress changes up to the end of RP1 are calculated for this plane. Events on F2 occurred during both reactivation periods, so stress changes are calculated for this plane for the entire duration of both reactivation periods. In the model, the receiver geometries were those of the individual fault planes, and receiver rake was given by the average rake of events used to Previous work in this ﬁeld (e.g. Catalli et al. 2013; Pennington & Chen 2017), observe CIs generally in excess of 70 per cent. Given the above results, elastic deformation may have only mod- estly promoted slip in this case, during the initial period of activity. However, during the latter period, the events were generally inhib- ited by the preceding elastic deformation. This implies that ﬂuid processes such as aseismic tensile slip, pore pressure changes and poroelastic effects were most likely the dominant cause of fault reactivation. These results are only slightly altered when the secondary nodal plane is chosen, with a CI of 61 per cent positive for the ﬁrst period and 36 per cent for the second. The windowed CI values also show Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 201 1208 1208 T. Kettlety et al. Figure 6. The values of the CFS for the two reactivation periods, using the preferred NP. Events receiving positive Coulomb stress change are shown in red, and negative in blue. Events with MW > 0 are shown as yellow stars. Note that a number of the larger magnitude events have higher magnitude, positive CFS. The CI is estimated across moving windows each containing 50 events, and shown by green diamonds connected with a dotted line. The green lines show the CI for the entire period. Above, bars indicate the duration of injection plotted over the same time periods. T. Kettlety et al. Figure 6. The values of the CFS for the two reactivation periods, using the preferred NP. 3.2 Resolving stresses onto fault structures Events receiving positive Coulomb stress change are shown in red, and negative in blue. Events with MW > 0 are shown as yellow stars. Note that a number of the larger magnitude events have higher magnitude, positive CFS. The CI is estimated across moving windows each containing 50 events, and shown by green diamonds connected with a dotted line. The green lines show the CI for the entire period. Above, bars indicate the duration of injection plotted over the same time periods. pressure effects must have dominated for F2, counteracting the ef- fect of inhibiting elastic CFS. This also highlights that, even when the uncertainty of resolving stresses onto the individual receiver ge- ometries is removed, roughly the same behaviour holds. However, geometry uncertainty is obviously not removed entirely here, as the sources of deformation are still the catalogued events. deﬁne the two fault planes (see Section 2.1). Cumulative CFS was calculated in half-hour increments, accumulating sources through successive time windows. The resulting stress changes acting at a given time on the plane are then calculated at sample points grid- ded at 20 m intervals along the surface. This grid was deﬁned by examining the extent of the cloud of seismicity. F1 extended for 180 m along its strike and 500 m in the dip direction. F2 extended 270 m along its strike, and 550 m in the dip. The fault dimen- sions can be seen in Fig. 4. These gridded stresses can then be analysed in a similar manner to that of an event population, by examining the proportion of positive CFS received (Coulomb in- dex), or the sum, or average of the stress changes at the sample points. 3.3 Sensitivity analysis The results of mapping elastic Coulomb stress changes onto the fault planes shown in Fig. 4. Grey bars show the number of sources that occurred in each of the 30 min time bins. Green lines show the effective CI—the proportion of points across the planes which are experiencing positive CFS at the time bin shown. Red show the sum of CFS across every sample point on the planes. Blue show the average CFS at the sample points. gj Stress transfer during hydraulic fracturing 1209 Figure 7. The results of mapping elastic Coulomb stress changes onto the fault planes shown in Fig. 4. Grey bars show the number of sources that occurred in each of the 30 min time bins. Green lines show the effective CI—the proportion of points across the planes which are experiencing positive CFS at the time bin shown. Red show the sum of CFS across every sample point on the planes. Blue show the average CFS at the sample points. Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 7. The results of mapping elastic Coulomb stress changes onto the fault planes shown in Fig. 4. Grey bars show the number of sources that occurred in each of the 30 min time bins. Green lines show the effective CI—the proportion of points across the planes which are experiencing positive CFS at the time bin shown. Red show the sum of CFS across every sample point on the planes. Blue show the average CFS at the sample points. Figure 8. Coulomb index distributions for the subset of events used to investigate the effect of FM uncertainty. The vertical lines indicate the CIs when the preferred and secondary NPs are chosen from the original FM. The red bars shows the distribution for catalogues when the NP is chosen randomly from the original FM. The green bars show the catalogues where FMs are permuted by Von Mises distributed rotation angles. The blue bars show the catalogues with randomly oriented FMs. The horizontal lines show the standard deviations of the distributions, with squares indicating the mean CI. Figure 8. Coulomb index distributions for the subset of events used to investigate the effect of FM uncertainty. The vertical lines indicate the CIs when the preferred and secondary NPs are chosen from the original FM. 3.3 Sensitivity analysis The similarities between the two methods of stress change deter- minations continue for F2 through RP1 and RP2. The CI across the second plane never exceeded 50 per cent and rarely went above 30 per cent, indicating that across the surface of this plane, the majority of the elastic stress changes were inhibiting failure throughout both reactivation periods. The average and total stress change magnitudes acting on the two fault planes mirror these trends, with consistently positive CFS for F1 and negative for F2. The results of the sensitivity analysis are shown in Fig. 8. For this subset of events, the CI when using the preferred nodal plane is 64 per cent, and when using the secondary NP, are 63 per cent. When the nodal plane are randomly selected for each event, the CI values Thus, we reach the same conclusion – deformation may have modestly contributed to the failure on F1, but ﬂuid and/or pore Stress transfer during hydraulic fracturing 1209 Figure 7. The results of mapping elastic Coulomb stress changes onto the fault planes shown in Fig. 4. Grey bars show the number of sources that occurred in each of the 30 min time bins. Green lines show the effective CI—the proportion of points across the planes which are experiencing positive CFS at the time bin shown. Red show the sum of CFS across every sample point on the planes. Blue show the average CFS at the sample points. Figure 8. Coulomb index distributions for the subset of events used to investigate the effect of FM uncertainty. The vertical lines indicate the CIs when the preferred and secondary NPs are chosen from the original FM. The red bars shows the distribution for catalogues when the NP is chosen randomly from the original FM. The green bars show the catalogues where FMs are permuted by Von Mises distributed rotation angles. The blue bars show the catalogues with randomly oriented FMs. The horizontal lines show the standard deviations of the distributions, with squares indicating the mean CI. are approximately normally distributed around a mean of 63±1 per cent This indicates that the choice of NP does not have a signiﬁcant random event geometries gave CI values distributed around 50±3 per cent as expected for entirely randomly aligned fracture sets Stress transfer during hydraulic fracturing 1209 Stress transfer during hydraulic fracturing 1209 Figure 7. 3.4 Cluster stress budget To investigate the effect of tight clustering on the elastic stress change budget, the DBSCAN algorithm (Ester et al. 1996) is used again to identify clusters within the event population. Whereas in Section 3.1 we used only spatial dimensions to initially identify fault reactivation, we now incorporate time as an additional dimension for the clustering analysis. A scaling factor of c = 250 ms−1 is applied to the time dimension. This value is found to best separate the clusters in the time domain. Following the method of Daszykowski et al. (2001), an optimal neighbourhood distance is calculated using the sorted k-distance approach, giving a value of 35 m. The resulting clusters are shown in Fig. 10. Fig. 14, shows the result of the probabilistic analysis using Fault- SlipPotential, showing the cumulative density function for the prob- ability of slip given some pore pressure perturbation P for F1 and F2. The probability of failure does not exceed 50 per cent until the pore pressure change is in excess of 15 MPa for both of the fault planes. F1 is marginally more optimally aligned for failure, with F2 requiring an additional ∼1 MPa of pore pressure change to give the same probability of failure. This scale of pore pressure change (∼10 MPa) is similar to that found by Chiaramonte et al. (2007), for a trap bounding fault in north-central North America. Across the range of input values, the SHmin and in situ pore pressure uncertainties had the largest effect on the P to slip for both faults. Signiﬁcantly re- ducing SHmin will have the effect of shifting the Mohr circle towards the failure criterion. Increasing the in situ pore pressure will act to decrease the effective stresses acting to clamp the fault. We calculate the stress changes in the same manner as Section 3.1, however, for each receiver, only sources from within the same cluster are used. This will allow for the full cumulative stress change to be compared to the change when only the deformation from within each cluster is considered. These resulting stress change value is termed the ‘intracluster’ stress change. A P required to fail of approximately 15 MPa would indi- cate that the two hypothesized faults here are not critically stressed within their stress ﬁeld. A large pore pressure increase would be re- quired for the Mohr–Coulomb failure criterion to be reached. 3.3 Sensitivity analysis The red bars shows the distribution for catalogues when the NP is chosen randomly from the original FM. The green bars show the catalogues where FMs are permuted by Von Mises distributed rotation angles. The blue bars show the catalogues with randomly oriented FMs. The horizontal lines show the standard deviations of the distributions, with squares indicating the mean CI. are approximately normally distributed around a mean of 63±1 per cent. This indicates that the choice of NP does not have a signiﬁcant impact on the resulting Coulomb index when a uniform slip model is used. When the FMs are rotated by Von Mises distributed angles, the mean CI is decreased to 59±3 per cent. Simulating entirely random event geometries gave CI values distributed around 50±3 per cent, as expected for entirely randomly aligned fracture sets. As this perturbation in fault geometry is applied, the CI distri- bution shifts toward that of the random focal mechanism scenario. A FM uncertainty of σ = 30◦would appear to correspond to an 1210 T. Kettlety et al. One would expect this to be true for clusters 1 and 2, where few events outside of the cluster preceded the seismicity, but this lin- ear relationship is also present for cluster 5 (Fig. 12). This cluster will have received all of the CFS from all of those proceeding it, but still, the majority of the stress changes comes from within that cluster. This agrees with Vasudevan & Eaton (2011), in that for microseismic events, where the spacial propagation of stress is not laterally extensive, the majority of the stress change comes from nearby sources. One would expect this to be true for clusters 1 and 2, where few events outside of the cluster preceded the seismicity, but this lin- ear relationship is also present for cluster 5 (Fig. 12). This cluster will have received all of the CFS from all of those proceeding it, but still, the majority of the stress changes comes from within that cluster. This agrees with Vasudevan & Eaton (2011), in that for microseismic events, where the spacial propagation of stress is not laterally extensive, the majority of the stress change comes from nearby sources. approximately ±4 per cent uncertainty in the observed CI values. However, even when the focal mechanisms are rotated by around 30◦, the positive signal observed during RP1 is still present. As shown in Catalli et al. 3.5 Fault slip potential The code FaultSlipPotential (Walsh et al. 2017) is used to con- duct a probabilistic geomechanical analysis, using the methodology described in Walsh & Zoback (2016). The stress gradients, SHmax azimuth, fault geometries, and the friction coefﬁcient are used as inputs, as well as their associated uncertainties. The principal stress uncertainties used are those given in Section 2.2, along with a μ of 0.7±0.1. The uncertainty in the orientation of the faults is given in Section 2.1. These input parameters are randomly sampled 1000 times within their uncertainties, and a resulting critical pore pressure required to initiate failure, PC, is then calculated using Coulomb’s law: Varying Skempton’s ratio appears to have a similar scale of effect on the CI as applying rotations to the event geometries (Fig. 9). CI changes from 64 per cent to 56 per cent, as β varies from 0 to 1, simulating entirely unsaturated to saturated faults. Note that when β > 0.4, CI decreases at a more rapid rate than when 0 < β < 0.4. When looking at the values of the individual event CFS, changing β generally acted to change the magnitude, but infrequently changed the sign – only 14 per cent of receivers changed sign across the whole range of β. This minority are responsible for the change in CI as measured for the population. The sign of all receivers in this sample with \|CFS\| > 0.05 MPa remained constant from one extreme β to the other. This indicates that if injection ﬂuids are interacting with the faults near the injection points (increasing β from 0.4) the value of the CI may decrease up to approximately 4 per cent. This would weaken the positive signal observed during the ﬁrst reactivation period. (3) PC = σn −\|τ\|/μ. (3) PC = σn −\|τ\|/μ. For the range of PC perturbations, a cumulative probability of ex- ceeding the Mohr-Coulomb failure criterion is then found. Fault- SlipPotential uses uniform distributions from which to sample the input parameters. This will give a narrower distribution in the re- sulting PC values, than would be found when using standard errors. We use eq. (3) to examine the in situ stress ﬁeld with respect to the fault orientations (shown in Fig. 4), as delineated by the microseismicity. Fig. 13 shows the computed σ n, τ, and PC as a function of fault strike and dip. 3.5 Fault slip potential Optimally oriented faults, where PC is at its minimum value of 14.5 MPa, have strikes of 28◦, 81◦, 208◦and 261◦. These are relatively close to the strikes of the planar structures: 250◦and 60◦. 3.3 Sensitivity analysis (2013), any perturbation to the FMs appears to systematically decrease the resulting average CI. Meier et al. (2014) interpreted this result as the outcome of applying further randomness to an already uncertain, noisy FM catalogue. We see the same result here. This challenge of quantifying the effect of focal mechanism uncertainty, and the signiﬁcance of the signals observed, is further discussed in Section 4. With injection ﬂuids interacting with the proposed faults in this system, the effect of changing pore pressure cannot be neglected. Thus, further sensitivity analysis is conducted to examine the effect of varying Skempton’s ratio β. Using the same subset of events as in the above FM analysis, stress changes are repeatedly modelled with a varying β. Values of μ ′ from 0 and 0.7 are used, in 0.1 increments, which corresponds to Skempton’s ratio varying from 0 ≤β ≤1. 3.4 Cluster stress budget With the average observed elastic stress increase on the fault planes being approximately 0.1 MPa, these planes would require two orders of magnitude larger pore pressure changes to initiate failure. Even at the extremes of the stress model and fault parameter uncertainty, the required stress change for failure is an order of magnitude Fig. 11 shows that, for the majority of events (89 per cent), the intracluster CFS makes up effectively all of the stress change received. This indicates the importance of tight clustering for mi- croseismic stress transfer, as even when sources are excluded within one source length, the stress change from within the cluster con- tributes the majority of the CFS. The relationship in Fig. 11 also shows that when CFS is signiﬁcant (i.e. greater than ±0.1 MPa), effectively all of the stress change results from within the cluster. Stress transfer during hydraulic fracturing 1211 Figure 9. The CI change for the ﬁrst 500 events in the ﬁrst activation period, with a varying value of Skempton’s ratio β. (a) The CI–β relationship for all the of events in the sample. (b) The temporal change in CI, calculated using moving windows, each containing 50 events. The grey-scale lines show the values of CI for the varying β values, while the green triangles show the mean CI for that time. The error bars give the standard deviation. The green dashed line shows the mean CI for the entire sample of events. Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 9. The CI change for the ﬁrst 500 events in the ﬁrst activation period, with a varying value of Skempton’s ratio β. (a) The CI–β relationship for all the of events in the sample. (b) The temporal change in CI, calculated using moving windows, each containing 50 events. The grey-scale lines show the values of CI for the varying β values, while the green triangles show the mean CI for that time. The error bars give the standard deviation. The green dashed line shows the mean CI for the entire sample of events. Figure 10. (a) Locations of the ﬁve clusters determined by the DBSCAN algorithm. (b) Temporal distribution of the clusters. d by the DBSCAN algorithm. (b) Temporal distribution of the clusters. Figure 10. (a) Locations of the ﬁve clusters determined by the DBSCAN algorithm. (b) Temporal distribution of the clusters. 3.4 Cluster stress budget faults, with very little intermediate seismicity. This would also sug- gest a rapid transport of ﬂuid along the faults, and then subsequent failure back up along the plane of weakness. This type of behaviour has been observed in EGS-induced microseismic sequences (Majer et al. 2007), and is described as a potential factor in the maximum observed magnitude of the injection induced seismicity (Shapiro et al. 2011). More complex ﬂuid and fracture modelling is required to fully examine the pressure changes occurring within the fault system, however this slip potential model does indicate that these faults are not optimally oriented for failure in the regional stress state. above the average elastic CFS. This suggests that the interac- tion with high pressure injection ﬂuids was likely to have been responsible for the activity in both reactivation periods, and elastic stress transfer did not play a major role in the fault activation. As discussed in Section 2.2, this analysis does not take into account the potential for overpressure in the formation, other than in the uncertainty applied to the in situ P (±7 MPa). During injection downhole pressures were measured to be consistently greater than 50 MPa, giving a differential pressure between that and the in situ pore pressure of over 20 MPa. Given the top of F1 is ∼50 m from the nearest perforation, and the fact that fractures are likely to ex- tend at least that distance, it is plausible that the pressurized ﬂuid interacted with the pre-existing structure and exceeded this failure criterion. above the average elastic CFS. This suggests that the interac- tion with high pressure injection ﬂuids was likely to have been responsible for the activity in both reactivation periods, and elastic stress transfer did not play a major role in the fault activation. As discussed in Section 2.2, this analysis does not take into account the potential for overpressure in the formation, other than in the uncertainty applied to the in situ P (±7 MPa). During injection downhole pressures were measured to be consistently greater than 50 MPa, giving a differential pressure between that and the in situ pore pressure of over 20 MPa. Given the top of F1 is ∼50 m from the nearest perforation, and the fact that fractures are likely to ex- tend at least that distance, it is plausible that the pressurized ﬂuid interacted with the pre-existing structure and exceeded this failure criterion. 4 D I S C U S S I O N This analysis however uses a simple model of fault stability, and does not take into account the possibility of frictional asperities on the fault surface. These areas of weakness would allow the failure threshold to be reached on a small fraction of the fault surface and enable failure to propagate outwards. This type of simple modelling also does not explain the spatiotemporal clustering observed here – small events near the well start the sequence, but immediately after and 400 m below, larger magnitude events occur at the base of the 4.1 Elastic stress modelling The positive signal we observe in the ﬁrst period of activity is a CI of around 60 per cent, as well as small periods wherein CI reached around 70 per cent. Ascribing signiﬁcance to this signal, a slightly higher proportion of events receiving positive CFS, is dif- ﬁcult, especially given the uncertainties in the FMs and the assump- tions inherent in modelling elastic stress change. We see similar T. Kettlety et al. 1212 Figure 11. (a) The relationship between the total CFS from all possible sources and the stress change due to sources from only within the same cluster. Percentages break down the proportions of events (865 within clusters of a possible 1064) in each octant. For example, the NNE octant indicates 22 per cent of events have a positive cluster CFS equal to or greater than the total CFS experienced from all possible sources. (b) The difference in intracluster stress change and total CFS. It is clear that for the vast majority of events (97 per cent), intracluster CFS makes up the majority of the stress change received. The absolute difference exceeds ±0.05 MPa for only 27 of 865 events. Downloaded from https://academic.oup.com/gji/article-abstract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 11. (a) The relationship between the total CFS from all possible sources and the stress change due to sources from only within the same cluster. Percentages break down the proportions of events (865 within clusters of a possible 1064) in each octant. For example, the NNE octant indicates 22 per cent of events have a positive cluster CFS equal to or greater than the total CFS experienced from all possible sources. (b) The difference in intracluster stress change and total CFS. It is clear that for the vast majority of events (97 per cent), intracluster CFS makes up the majority of the stress change received. The absolute difference exceeds ±0.05 MPa for only 27 of 865 events. Figure 12. Cluster stress change against total stress change for each of the ﬁve clusters. Percentages break down the proportions of events in each octant, as in Fig. 11(a). Clusters 1 and 4 receive very little stress change, as they are low magnitude, broadly clustered events associated with the fracturing. tract/217/2/1200/5315764 by University of Bristol Library user on 14 March 2019 Figure 12. Cluster stress change against total stress change for each of the ﬁve clusters. 4.1 Elastic stress modelling Percentages break down the proportions of events in each octant, as in Fig. 11(a). Clusters 1 and 4 receive very little stress change, as they are low magnitude, broadly clustered events associated with the fracturing. Stress transfer during hydraulic fracturing 1213 Figure 13. Modelled HRB in situ stresses resolved onto fault planes, assuming a coefﬁcient of friction of 0.7. (a) Effective normal stress. (b) Shear stress. (c) Critical pore pressure required to reach the Mohr failure criterion. The resolved stresses are contoured at the polar projection of the normal to the fault plane, with polar angle representing the fault-normal trend, and radial angle representing the fault-normal plunge. The orientations of the two faults F1 and F2 are shown respectively by the square and the triangle, both close to the minimum PC values. Stress transfer during hydraulic fracturing 1213 Stress tra Figure 13. Modelled HRB in situ stresses resolved onto fault planes, assuming a coefﬁcient of friction of 0.7. (a) Effective normal stress. (b) Shear stress. (c) Critical pore pressure required to reach the Mohr failure criterion. The resolved stresses are contoured at the polar projection of the normal to the fault plane, with polar angle representing the fault-normal trend, and radial angle representing the fault-normal plunge. The orientations of the two faults F1 and F2 are shown respectively by the square and the triangle, both close to the minimum PC values. signal is still present. This indicates that despite FM uncertainty, the events in the ﬁrst reactivation period are spatially distributed in such a way that elastic stress changes still modestly promote failure. Figure 14. Fault slip potential probability curves for the two modelled faults. The inherent assumptions used and lack of data must also be considered, and their effect on the accuracy of our results. Both observational precision, and lack of data, constrain the accuracy of our modelling. Uncertainties in both the FMs and locations were not provided by the processing company, and MTs were not com- puted for a number of events in the original catalogues in this study due to low signal-to-noise ratios. That said, these ‘missing’ events generally have considerably lower magnitudes (MW < −1.5) than the majority of events used here (those with MTs), so the effect on the stress change from these sources is expected to be small. 4.1 Elastic stress modelling Their contribution to the stress changes will therefore be minimal and is assumed to not have a signiﬁcant effect on the results. The uniform slip model used, which gives rise to a number of the limitations described above (NP ambiguity, source exclusion within 1 source length), has a dramatic effect on the accuracy of our results, as it does for all Coulomb stress modelling. A fuller treatment of modelling slip distributions, however, would have been impractical in this case, with up to ∼1400 sources being modelled for later receivers. Also, the nature of slip dis- tribution on the fault plane is highly uncertain in itself. Ascrib- ing some assumed rupture behaviour in the hope of more ac- curately reﬂecting the near-ﬁeld stress change could in fact pro- duce signiﬁcantly more uncertain CFS values within one source length. Figure 14. Fault slip potential probability curves for the two modelled faults. magnitudes of stress changes as previous studies (e.g. Schoenball et al. 2012), however our results diverge from studies which ﬁnd CIs in excess of 75 per cent, implying that elastic stress transfer played a more signiﬁcant role during injection induced seismicity (e.g. Catalli et al. 2013; Pennington & Chen 2017). We have also tested the reliability of this signal by conducting a separate anal- ysis, wherein individual receiver geometry is bypassed, and stress changes are instead mapped onto the visible fault planes. Whilst the measured quantities are different, the same pattern emerges – a positive signal in the ﬁrst reactivation period, and a generally negative signal in the second. Testing the sensitivity to particular uncertainties conventionally, using the method employed in Catalli et al. (2013), we observe a decrease in CI by both the focal mecha- nism (∼4 per cent) and frictional uncertainty (∼4 per cent), which weaken the positive signal observed. However, this approach may not be a reﬂection of the effect of FM uncertainty on the resulting CI. As described in Meier et al. (2014), adding random noise to the FMs by applying a perturbation just adds a smearing effect, always resulting in a CI closer to 50 per cent, no matter how signiﬁcant the original signal. This method however, does show that even when FMs are rotated by, on average, a very signiﬁcant amount (30◦), the magnitudes of stress changes as previous studies (e.g. Schoenball et al. 4.2 Focal mechanism sensitivity analysis the symmetrical nature of the stress change pattern that results from a uniform slip model. Applying Von Mises distributed uncertainties to the focal mechanisms of the events appears to diminish, but does not completely eliminate, the observed CI signals. However, this is due to the nature of measuring the FM sensitivity on a population of events with already uncertain FMs, which acts to consistently shift the CI towards 50 per cent. Variability in the Skempton’s ratio is also found to diminish, but not completely eliminate, the positive CI signal. The scale to which the resulting CI distributions shift while varying the model parameters highlights the importance of properly incorporating the various uncertainties that affect CFS calculations. Comparing our permuted focal mechanism catalogues to those with entirely random FMs (Fig. 8) also may not provide an accurate mea- sure of signiﬁcance when considering a possible null hypothesis, as described by Meier et al. (2014). A more suitable method could involve comparing to the CFS computed for events not thought to be triggered by elastic stress changes, and thus not associated with the fault activation. This could include populations near to the wells, assumed to be caused by tensile fracturing. For these events, ﬂuid effects are expected to dominate and no considerable elastic triggering, that is, the CI for this population would be around 50 per cent. However, the accuracy of the focal mechanisms for these events would be even more questionable, with event size generally around one to two magnitudes smaller than the fault reactivation events. We already have considered a relatively pessimistic uncer- tainty for the FMs (30◦), however, this would most likely increase dramatically for those events where the signal-to-noise ratio is even smaller. Another consideration is the alignment of failure planes for tensile fracturing events. These events will be generally well aligned parallel to the maximum stress direction, and so their rel- ative alignments will deviate from that a zone of random oriented fractures. Thus, due to the spatial pattern of the stress change, they will predominantly receive positive stress change, and static stress transfer will appear to play a signiﬁcant role. Intracluster stress transfer is found to be signiﬁcant in this case, meaning that the elastic stress change budget is chieﬂy derived from events in the same cluster. AC K N OW L E D G E M E N T S The authors would like to thank the operator for supplying the data, and ESG Solutions Ltd., the service provider who collected and processed the data set. TK was supported by the Natural Environ- ment Research Council (NERC) GW4+ Doctoral Training Partner- ship (grant number NE/L002434/1). MJW was supported by NERC (grant number NE/R017956/1) and the Southern California Earth- quake Center (SCEC) (Contribution No. 8961). SCEC is funded by NSF Cooperative Agreement EAR-1600087 and USGS Coopera- tive Agreement G17AC00047. JPV was supported by NERC (grant number NE/R018162/1). JMK was supported by NERC (grant num- ber NE/R018006/1). This project was supported in part by Bristol University Microseismicity Projects (BUMPs). This work used data from the World Stress Map Database Release 2016 of Heidbach et al. (2016) . The code MTDecompostion of Vavryˇcuk (2015a) and MATLAB scripts produced by Meier et al. (2014) were both mod- iﬁed for use in this work. The DBSCAN algorithm of Ester et al. (1996) was used to determine spatiotemporal clusters of events. The code FaultSlipPotentialv1.07 of Walsh et al. (2017) was used in the latter sections. We would like to thank the reviewers for their very constructive feedback, which improved the quality of the manuscript. This could provide a possible explanation as to the difference in signiﬁcance of elastic CFS seen in this study compared to other sites (e.g. Catalli et al. 2013; Sumy et al. 2014; Penning- ton & Chen 2017). If the faults are already close to being crit- ically stressed, then the small amounts of stress change associ- ated with static elastic stress transfer can play a signiﬁcant role. If the faults are not close to failure, then larger stress changes are required. In such cases the stress changes associated with static elastic stress transfer are not large enough, and instead the very high pressures associated with ﬂuids dominate the activation of faults, meaning that static stress transfer does not play a signiﬁcant role. 4.3 Fault orientations The geometry of the mapped faults and an estimate of the in situ stress regime indicate that the activity seen here was initiated by the high-pressure injection near the failure plane, as faults require in excess of 10 MPa of pore pressure change to reach the Mohr failure criterion. This is in contrast to the stress environment and fault system found in central the US, where waste-water injection has stimulated high magnitude (>3 MW) events regularly in recent years (Alt & Zoback 2017). Both Walsh & Zoback (2016) and Schoenball et al. (2018) found, using the QRA methods employed here, that multiple fault systems mapped in north-central Oklahoma are preferentially aligned for failure, with some faults requiring only ∼2 MPa of P to reach a failure potential of 0.5. In our case, the required pore pressure changes are approximately six times greater. 4.1 Elastic stress modelling 2012), however our results diverge from studies which ﬁnd CIs in excess of 75 per cent, implying that elastic stress transfer played a more signiﬁcant role during injection induced seismicity (e.g. Catalli et al. 2013; Pennington & Chen 2017). We have also tested the reliability of this signal by conducting a separate anal- ysis, wherein individual receiver geometry is bypassed, and stress changes are instead mapped onto the visible fault planes. Whilst the measured quantities are different, the same pattern emerges – a positive signal in the ﬁrst reactivation period, and a generally negative signal in the second. Testing the sensitivity to particular uncertainties conventionally, using the method employed in Catalli et al. (2013), we observe a decrease in CI by both the focal mecha- nism (∼4 per cent) and frictional uncertainty (∼4 per cent), which weaken the positive signal observed. However, this approach may not be a reﬂection of the effect of FM uncertainty on the resulting CI. As described in Meier et al. (2014), adding random noise to the FMs by applying a perturbation just adds a smearing effect, always resulting in a CI closer to 50 per cent, no matter how signiﬁcant the original signal. This method however, does show that even when FMs are rotated by, on average, a very signiﬁcant amount (30◦), the Thus, we followed the methodology of previous works (such as Meier et al. 2014), where slip is treated as uniform, and near-ﬁeld sources are ignored. This method obviously results in an lower es- timate for the resulting CFS magnitude, but also removes the signiﬁcant effect of the uncertainty in the stress drop, slip model, and to a certain extent source location (Steacy et al. 2004; Schoen- ball et al. 2012). These parameters, each of which have signiﬁcant uncertainties, mostly effect the near-ﬁeld stress change, due to the r−3 power law describing the decay of CFS. The resulting values can thus be considered a more accurate reﬂection of the triggering effect of elastic stress change. One could imagine that this exclu- sion step is ampliﬁed in this study in particular, where events are relatively tightly clustered. However, as shown in Fig. 12, the CFS from within clusters still makes up a signiﬁcant proportion of the stress change received, and thus effect from sources within a cluster is not negated using this method. 1214 T. Kettlety et al. 4.2 Focal mechanism sensitivity analysis This is expected for tightly clustered microseismic events because of the small area and (size of) slip on the fault plane, resulting in spatially contained elastic stresses. Thus, events only receive signiﬁcant CFS when they are within ∼100 m from the source. The weak contribution of elastic stress transfer to the failure can be interpreted with respect to the in situ stresses acting on the hypothesized fault structures. Using at-depth measurements of the regional principal stresses, the probability of slip for a given pore pressure change on the two planar structures is examined using a quantitative risk assessment technique. We ﬁnd that for both faults, the pore pressure changes required to reach the failure criterion are around 15 MPa, far in excess of the reliable values of elastic stress change. This supports that, in this case, the failure was stimulated by the presence of highly pressurized ﬂuids in the system. These faults are not critically stressed, unlike those found in the central US, which have been found to require only a few MPa of pore pressure to reach the slip criterion (Walsh & Zoback 2016; Schoenball et al. 2018). This could provide an explanation for the difference between the CFS behaviour found here and studies which concluded that this mechanism played a more signiﬁcant role (Catalli et al. 2013; Pennington & Chen 2017). R E F E R E N C E S Goertz-Allmann, B.P., Goertz, A. & Wiemer, S., 2011. Stress drop variations of induced earthquakes at the Basel geothermal site, Geophys. Res. Lett., 38(9), L09308, doi:10.1029/2011GL047498. Goertz-Allmann, B.P., Goertz, A. & Wiemer, S., 2011. Stress drop variations of induced earthquakes at the Basel geothermal site, Geophys. Res. Lett., 38(9), L09308, doi:10.1029/2011GL047498. Aki, K., 1965. Maximum likelihood estimate of b in the formula log N =a −bm and its conﬁdence limits, Bull. Earthq. Res. Inst., 43, 237–239. 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https://openalex.org/W2737115268	https://hal.archives-ouvertes.fr/hal-01567396/document	English	null	Hybrid control for low-regular nonlinear systems: application to an embedded control for an electric vehicle	arXiv (Cornell University)	2,017	cc-by-sa	12,818	Hybrid control for low-regular nonlinear systems: application to an embedded control for an electric vehicle Thomas Chambrion, Gilles Millérioux To cite this version: Thomas Chambrion, Gilles Millérioux. Hybrid control for low-regular nonlinear systems: application to an embedded control for an electric vehicle. 2017. hal-01567396 HAL Id: hal-01567396 https://hal.science/hal-01567396v1 Preprint submitted on 23 Jul 2017 L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. HAL is a multi-disciplinary open access archive for the deposit and dissemination of sci- entific research documents, whether they are pub- lished or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. Distributed under a Creative Commons Attribution - ShareAlike 4.0 International License Hybrid control for low-regular nonlinear systems: application to an embedded control for an electric vehicle Thomas Chambrion∗†‡ Gilles Millerioux§ July 23, 2017 Abstract This note presents an embedded automatic control strategy for a low consumption vehicle equipped with an “on/oﬀ” engine. The main diﬃculties are the hybrid nature of the dynamics, the non smoothness of the dynamics of each mode, the uncertain environment, the fast changing dynamics, and low cost/ low consumption constraints for the control device. Human drivers of such vehicles frequently use an oscillating strategy, letting the velocity evolve between ﬁxed lower and upper bounds. We present a general justiﬁcation of this very simple and eﬃcient strategy, that happens to be optimal for autonomous dynamics, robust and easily adaptable for real-time control strategy. Eﬀective implementation in a competition prototype involved in low-consumption races shows that automatic velocity control achieves performances comparable with the results of trained human drivers. Major advantages of automatic control are improved robustness and safety. The total average power consumption for the control device is less than 10 mW. ss and safety. The total average power consumption for the control device is less than 10 mW. Contents Contents 1 Introduction 2 2 Autonomous dynamics in ﬁnite time 3 2.1 Statement of the optimal control problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 2.2 Preliminaries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2.3 Physical assumptions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.4 Oscillating structure of the solutions of Problem 1 . . . . . . . . . . . . . . . . . . . . . . . . . 7 3 Autonomous dynamics with inﬁnite time horizon 9 3.1 Statement of the optimal problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.2 Main Result . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.3 Proof . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.4 Practical determination of the upper and lower speed limits . . . . . . . . . . . . . . . . . . . . 12 4 Non autonomous problem in ﬁnite time 12 4.1 Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . †CNRS, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France ‡ ∗Universit´e de Lorraine, IECL, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France † ‡Inria, 54600 Villers, France, Thomas.Chambrion@univ-lorraine.fr § , , e Lorraine, CRAN, UMR 7039, 54506 Vandoeuvre-l`es-Nancy, France, Gilles.Millerioux@univ-lorraine.f Contents . . . . . . . . . . . . . . . . . . . . . . 12 4.2 Description of the strategy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 4.3 Well-posedness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 4.4 Constraints fulﬁllment and robustness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 5 Eﬀective implementation 14 5.1 Vir’Volt 2 prototype . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 5.2 Eﬀective results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 6 Conclusions 15 ∗Universit´e de Lorraine, IECL, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France †CNRS, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France ‡Inria, 54600 Villers, France, Thomas.Chambrion@univ-lorraine.fr §Universit´e de Lorraine, CRAN, UMR 7039, 54506 Vandoeuvre-l`es-Nancy, France, Gilles.Millerioux@univ-lorraine.fr Co te ts 1 Introduction 2 2 Autonomous dynamics in ﬁnite time 3 2.1 Statement of the optimal control problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 2.2 Preliminaries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2.3 Physical assumptions . . . . . . . . . . Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2.4 Oscillating structure of the solutions of Problem 1 . . . . . . . . . . . . . . . . . . . . . . . . . 7 3 Autonomous dynamics with inﬁnite time horizon 9 3.1 Statement of the optimal problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.2 Main Result . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.3 Proof . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.4 Practical determination of the upper and lower speed limits . . . . . . . . . . . . . . . . . . . . 12 4 Non autonomous problem in ﬁnite time 12 4.1 Modeling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 4.2 Description of the strategy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 4.3 Well-posedness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . †CNRS, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France ‡ 502, 54506 Vandoeuvre-l`es-Nancy, France lers, France, Thomas.Chambrion@univ-lorraine.fr A §Universit´e de Lorraine, CRAN, UMR 7039, 54506 Vandoeuvre-l`es-Nancy, France, Gilles.Millerioux@un ∗Universit´e de Lorraine, IECL, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France †CNRS UMR 7502 54506 Vandoeuvre l`es Nancy France ∗Universit´e de Lorraine, IECL, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, Franc †CNRS, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France Contents . . . . 13 4.4 Constraints fulﬁllment and robustness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 5 Eﬀective implementation 14 5.1 Vir’Volt 2 prototype . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 5.2 Eﬀective results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 6 Conclusions 15 ∗Universit´e de Lorraine, IECL, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France †CNRS, UMR 7502, 54506 Vandoeuvre-l`es-Nancy, France ‡Inria, 54600 Villers, France, Thomas.Chambrion@univ-lorraine.fr §Universit´e de Lorraine, CRAN, UMR 7039, 54506 Vandoeuvre-l`es-Nancy, France, Gilles.Millerioux@univ-lorraine.fr 1 1 Figure 1: Test run of Vir’Volt 2 in Rotterdam (May 2014). Figure 1: Test run of Vir’Volt 2 in Rotterdam (May 2014). 1 Introduction The European Shell Eco-MarathonTM brings together over 200 teams from high schools and universities from all over Europe, in a race involving ecological and economical vehicles. The principle of the race is to go through a given distance in a limited time and with the lower energy consumption. The aim of this note is to expose the design and the implementation of an embedded automatic control of the speed on the prototype Vir’Volt 2 (see Figure 1), built by the students of ´Ecole Sup´erieure des Sciences et Technologies de Nancy (ESSTIN) in France that participated to the edition 2014 of the European Shell Eco-Marathon. A crucial feature of the prototype is the hybrid nature of the control. Indeed, the DC electric motor is either switched on or switched oﬀ. The energy consumption of the vehicle is reduced to a roughly constant residual consumption of the electronics (a few milliwatts) when the engine is oﬀand goes up to 160 W when the engine is on. When the engine is oﬀ, the transmission parts (gears, chains,...) are decoupled from the wheels and rapidly stops (the kinetic energy is dissipated by friction and Joule eﬀect in the engine). When the engine is switched on, the transmission parts (gears, chains, ..) that were at rest have to speed up in order to reach the wheels velocity, before the coupling allows energy to be transmitted from the engine to the wheels. The diﬀerence between the kinetic energy of the freely evolving transmission parts and the transmissions parts rotating at engine speed is far from being negligible (about 10 J), and has to be paid each time the motor is switched on. Clearly, the issue enters the context of control in switched systems. Switching systems are widely involved in industrial processes [23] and have been a major ﬁeld of interest in automatic control [7][19] for years. Optimal control of such switched systems is now pretty well understood for linear dynamics [25, 26, 14, 21]. For non linear dynamics, the case without switching cost has raised considerable attention, see for instance [13, 28, 5, 4, 18, 1]. The optimization problem is classically tackled with the Pontriaguin Maximum Principle (PMP), see for instance [2] for an intrinsic formulation. Non smooth extensions of the PMP have been developed to include the case of non smooth trajectories or nonzero switching costs [27, 24]. 1 Introduction Any proposed strategy to adapt the prototype speed will be an imperfect trade-oﬀbetween robustness, computational tractability and eﬃciency. In energy eﬃcient prototypes competitions, human drivers usually have an oscillating strategy, letting the velocity evolve between a lower and an upper limit. Our main result is a rigorous proof that, with physically reasonable assumptions, the long term optimal strategy for a stationary problem (wind and track slope remain constant) has exactly this pattern. Taking advantage of the particular structure of these solutions, this paper gives an explicit control strategy for the general problem (variable wind and slope) to adapt the ﬁnite horizon of the optimization scheme, with the following features: • Very low computational burden, implementable on low-cost embedded micro-controller for real-time automatic control; • Very low computational burden, implementable on low-cost embedded micro-controller for real-time automatic control; • Very low power consumption (less than 10 mW in average, about 15 J for the race); • Real time adaptability with respect to changes in dynamics, weather or traﬃc conditio • Optimality of the control law if the race conditions (wind and slope) are constant. The result applies also in the case (important in practice) where the dynamics is not L The result applies also in the case (important in practice) where the dynamics is not Lipschitz continuous. The layout of the paper is the following. In Section 2, the time independent model of the dynamics is presented and a ﬁnite-time horizon is considered. For this special case (autonomous dynamics corresponding to constant slope of the track, wind strength and direction), we prove (Proposition 3) that the optimal driving strategies exhibit a very simple behavior, where the speed oscillates, most of the time, between a maximum and a minimum without intermediate local extrema. In Section 3, we consider a long term version of the problem studied in Section 2, that is inﬁnite time horizon. Once again, the optimal strategy exhibits a very simple periodic pattern. In Section 4, a non autonomous model for the dynamics is considered by taking into account possible external disturbances like variable slope, wind, . . . . Some preliminary well-posedness mathematical results are introduced. Next, we use the strong geometric structure of the optimal strategies in the autonomous case (the ﬁnite horizon of the optimization scheme is the period of the optimal control computed in Section 2). Stability and robustness analyses completes this section. 1 Introduction p j Two diﬃculties prevent the direct use of these results in our case. The ﬁrst diﬃculty arises when modeling the dynamics. In the standard frame of Lipschitz continuous dynamics, a vehicle with switched oﬀengine on a ﬂat road will never be at rest because of the principle of non-intersection of solutions. A more realistic modeling (where cars can actually stop) requires to consider less regular dynamics, which usually give rise to non-trivial technical subtleties. This explain that this approach is rare in practice (with few exceptions, such as [17] for instance). ( [ ] ) The second diﬃculty is the heavy computational burden. Even in the standard Lipschitz continuous frame- The second diﬃculty is the heavy computational burden. Even in the standard Lipschitz cont 2 work, with especially adapted algorithm [15, 9, 10], some dozen of seconds of computations are needed with a powerful desktop computer to compute the optimal strategy of a 39 minutes run, and any perturbation (traﬃc, weather change,...) requires a new computation. Following the rules of the competition, this computation has to be done or stored in the vehicle (not oﬀboard). Even for races where the electric energy needed to do the computation is not incorporated in the total energy consumption, the battery weight has bad inﬂuence on the total consumption. Online integration of the PMP requires a strong multi-core processor (about 50 W on average, about 100 kJ for the race). Table of optimal trajectories may be pre-computed oﬄine and stored inboard. The main drawback of this approach is the cost of construction (pre-computation) of such a table - this includes a good knowledge of the geometry of the track and a reliable meteorological date base. This pre-computation process has to be redone for every diﬀerent race. Since global optimization seems out of reach, one may consider ﬁnite horizon optimization (as in Model Predictive Control). Even in this case, the computational burden is not to be neglected: a small linux computer with an average power of 5 W requires about 10 kJ for the race, to be compared with the 100 kJ needed to move the vehicle. Moreover, the choice of the horizon is usually diﬃcult because it impacts on both feasibility [3, 8, 20] and stability issues (see [11, 16, 6, 22] for computational aspects with invariant set-based approaches). 1 Introduction Finally, Section 5 is devoted to the results obtainef from a real-life implementation on the prototype during the competition. 2.1 Statement of the optimal control problem For every x0 in R, for every u in {0, 1}, the Cauchy problem (1) with initial condition x(0) = x0 admits a unique solution in positive time. The existence (but not the uniqueness) of continuously diﬀerentiable solutions to the Cauchy problem (1) with initial condition x0 at time t0 is a consequence of Assumption 1.1 and Peano theorem (see for instance [12], Theorem 2.1 page 10). The local uniqueness in positive times of the solutions (Assumption 1.2) is needed for physical reasons and ensures the continuity of the solutions with respect to the initial conditions ([12], Theorem 2.1 page 94). Notice that Assumption 1 is not enough in general to ensure neither the uniqueness in past times nor the diﬀerentiability of the solutions with respect to the initial conditions ([12], Theorem 6.1 page 104). 2.1 Statement of the optimal control problem Let us consider the unperturbed dynamics of the vehicle given by: ˙x = f(x, u) (1) (1) ˙x = f(x, u) where x is the real-valued speed of the prototype, u is the control, a piecewise constant function taking value in {0, 1} representing the state of the engine (0 is oﬀ, 1 is on), and f : R × {0, 1} →R is the dynamics. The initial condition x(0) = x0 ≥0 corresponds to the speed at start. 3 Hereafter, the total duration of the race will be denoted with T (T > 0), the length of the track will be denoted with L (L > 0) and the average speed will be denoted with V = L/T (V > 0). Let u : [0, T] →{0, 1} be piecewise constant control. For a real constant T1 > 0, a real-valued function x : [0, T1] →R is a solution of (1) if x is continuous, piecewise diﬀerentiable and satisﬁes (1) for almost every time t in [0, T1]. [ ] Let us recall that a real-valued function g : [0, T1] →R is absolutely continuous if g is diﬀerentiable almost everywhere in [0, T1] with integrable derivative g′ that satisﬁes g(t) = g(0) + R t 0 g′(s)ds for every t in [0, T1]. With every admissible control u and absolutely continuous solution x of (1), we associate the energy cost C(u) = Z T 0 h x(t), u(t) dt + α N(u) (2) (2) where h : [0, +∞) × {0, 1} →[0, +∞) is a C1 function accounting for the instantaneous power consummed by the vehicle, α is a positive real constant (representing the cost to switch the motor on) and N denotes the number of times that u jumps from 0 to 1. The problem to be solved is the following Problem 1 Find a piecewise constant control u : [0, T] →{0, 1} such that the associated solution of (1) with given initial condition x0 satisﬁes R T 0 x(t)dt = L and minimizes the cost (2). To ensure that the problem is well-posed, we need some regularity assumptions on f. Assumption 1 1. For any u in {0, 1}, the one-variable function x 7→f(x, u) is continuous. 2. 2.2 Preliminaries (8) (8) The above estimates are crucial for the variational arguments we will use in the proof of Pr The above estimates are crucial for the variational arguments we will use in the proof of Propositions 1 and 3. The above estimates are crucial for the variational arguments we will use in the proof of Propositions 1 and 3. 2.3 Physical assumptions We list below some mathematical assumptions, that correspond to reasonable physical assumptions. We list below some mathematical assumptions, that correspond to reasonable physical ass Assumption 2 1. The external actions like the slope of the track and the wind are not strong enough to prevent the car to go forward (maybe very slowly) when the engine is on: there exists V ∗> 0 such f(x, 1) > 0 if x < V ∗, f(V ∗, 1) = 0 and f(x, 1) < 0 if x > V ∗; 2. The (possibly negative) acceleration is larger when engine is switched on: for every x, f(x, 0) < f(x, 1); 3. When engine is oﬀ, the velocity tends to a (possibly negative) limit speed: there exists V∗such that f(x, 0) < 0 if x > V∗, f(x, 0) > 0 if x < V∗and f(V∗, 0) = 0. 4. Energy consumption is zero when the engine is oﬀ, positive when engine is on: for every x, h(x, 1) > h(x, 0) = 0; 4. Energy consumption is zero when the engine is oﬀ, positive when engine is on: for every x, h(x, 1) > h(x, 0) = 0; 5. Energy consumption increases with speed: x 7→h(x, 1) is non-decreasing; 5. Energy consumption increases with speed: x 7→h(x, 1) is non-decreasing; 5. Energy consumption increases with speed: x 7→h(x, 1) is non-decreasing; 6. The switching cost is not so large that the best strategy on long term is to keep full speed: all the integrals below converge and 6. The switching cost is not so large that the best strategy on long term is to keep full speed: all the integrals below converge and V ∗ Z V ∗ V∗ h(V ∗, 1) −h(s, 1) f(s, 1) ds + α < h(V ∗, 1) V ∗−V∗ Z V∗ V ∗ s −V∗ f(s, 0)ds + Z V ∗ V∗ s −V ∗ f(s, 1) ds ! ; 7. The energetic cost does not grow at the same rate than the engine eﬃciency when the velocity rises: the function F : x 7→ h(x,1)f(x,0) f(x,1)−f(x,0) is either strictly concave or strictly convex. 7. The energetic cost does not grow at the same rate than the engine eﬃciency when the velocity rises: the function F : x 7→ h(x,1)f(x,0) f(x,1)−f(x,0) is either strictly concave or strictly convex. 2.2 Preliminaries Let u : [t1, t2] ⊆[0, T] →{0, 1} be constant on the interval [t1, t2]. Then, the solution x : [t1, t2] →R of (1) with initial value x(t1) ⊆[0, L] is of class C1. If moreover t 7→f x(t), u(t) does not vanish on [t1, t2], then x(t) is increasing (if f x(t), u(t) > 0) or decreasing (if f x(t), u(t) < 0). As long as t 7→f(x(t), u(t)) does not vanish on (t1, t2), one can use the change of variable s = x in the following integrals and obtain expressions of the elapsed time between instants t1 and t2 t2 −t1 = Z t2 t1 dτ = Z x(t2) x(t1) ds f(s, u), (3) (3) of the covered length between instants t1 and t2 Z t2 t1 x(τ)dτ = Z x(t2) x(t1) sds f(s, u), (4) (4) and of the total energy consumption between instants t1 and t2: and of the total energy consumption between instants t1 and t2: Z t2 t1 h(x(τ), u(τ))dτ = Z x(t2) x(t1) h(s, u)ds f(s, u) . (5) (5) Indeed, the number of switches N(u) involved in (2) is zero in formula (5) since u is constant on [t1, t2]. From formulas (3), (4) and (5), one can notice that the speed average and the energy consumption are continuously 4 diﬀerentiable with respect to the piece-wise constant control u, as long as the acceleration does not vanish. In particular, for any given constant u, for any given V0, V1 and any w such that x 7→f(x, u) does not vanish on the convex hull of {V0, V1, V1 + w}, the diﬀerence between the time needed to reach speed V1 + w from V0 and the time needed to reach speed V1 from V0 expresses, at ﬁrst order, as Z V1+w V0 ds f(s, u) − Z V1 V0 ds f(s, u) = w f(V1, u) + ow→0(w). (6) (6) order variation of the covered length Similarly, one gets the ﬁrst order variation of the covered length Z V1+w V0 sds f(s, u) − Z V1 V0 sds f(s, u) = wV1 f(V1, u) + ow→0(w), (7) (7) and of the energy consumption and of the energy consumption Z V1+w V0 h(s, u)ds f(s, u) − Z V1 V0 h(s, u)ds f(s, u) = h(V1, u)w f(V1, u) + ow→0(w). 2.4 Oscillating structure of the solutions of Problem 1 Proposition 1 Under Assumptions 1 and 2, either Problem 1 has no solution or all of the solutions of Problem 1 have the following structure: with the possible exceptions of one initial and one ﬁnal acceleration/deceleration phase, the speed of the vehicle oscillates between two values without intermediate local extrema. Remark 1 Proposition 1 does not state that Problem 1 has a solution. It only states that any solution of Problem 1 has an oscillating structure (see Figure 3), which is no surprise. Indeed, if it is possible to write a Maximum Principle for this optimization problem, then the co-state and the associated switching function will be parametrized by the velocity x. As a general rule, there is a Maximum Principle for every optimization problem but a precise statement may require space and technicalities (see in particular the very stimulating discussion of [27, Section VI]). Notice also that the lack of backward uniqueness of trajectories prevents a direct use of the classical proofs of the Maximum Principle that can be found for instance in [2]. The rest of this section is dedicated to the proof of Proposition 1. The core of the proof is the following variational argument (Lemma 2) whose proof does not require the convexity guaranteed by Assumption 2.7. Lemma 2 Under Assumptions 1 and 2.1 to 2.5, the solutions (if any) of Problem 1 have the following struc- ture: with the possible exceptions of one initial and one ﬁnal acceleration/deceleration phase, the speed of the vehicle oscillates between local extrema V1, V2, . . . , Vp in [V∗, V ∗] and the matrix   1 1 . . . 1 V1 V2 . . . Vp h(V1)f(V1,0) f(V1,0)−f(V1,1) h(V2)f(V2,0) f(V2,0)−f(V2,1) . . . h(Vp)f(Vp,0) f(Vp,0)−f(Vp,1)   has rank at most two.   1 1 . . . 1 V1 V2 . . . Vp h(V1)f(V1,0) f(V1,0)−f(V1,1) h(V2)f(V2,0) f(V2,0)−f(V2,1) . . . h(Vp)f(Vp,0) f(Vp,0)−f(Vp,1)   has rank at most two. Proof of Lemma 2: Let u : [0, T] →{0, 1} be a piecewise constant optimal solution of Problem 1. The restriction to (0, T) of the associated trajectory x : [0, T] →R admits successive local extrema V1, V2, . . . , Vp in [V∗, V ∗] at points TV1, TV2, . . . 2.3 Physical assumptions Assumptions 2.1, 2.2, and 2.3 are summarized in Figure 2. The various terms involved in Assumption 2.6 can be interpreted in terms of velocity and accelerations. In particular, the ﬁrst term R V ∗ V∗ h(V ∗,1)−h(s,1) f(s,1) ds is equal to h(V ∗, 1) R V ∗ V∗ ds f(s,1) − R V ∗ V∗ h(s,1) f(s,1)ds, i.e., the opposite of the diﬀerence between the energy needed to accelerate from V∗to V ∗and the energy used by the vehicule running at full speed V ∗during the same amount of time R V ∗ V∗ ds f(s,1). The various terms involved in Assumption 2.6 can be interpreted in terms of velocity and accelerations. In particular, the ﬁrst term R V ∗ V∗ h(V ∗,1)−h(s,1) f(s,1) ds is equal to h(V ∗, 1) R V ∗ V∗ ds f(s,1) − R V ∗ V∗ h(s,1) f(s,1)ds, i.e., the opposite of the diﬀerence between the energy needed to accelerate from V∗to V ∗and the energy used by the vehicule running at full speed V ∗during the same amount of time R V ∗ V∗ ds f(s,1). 5 Time Time Speed Speed V ∗ V∗ Figure 2: Time plot of the dynamics with constant control u = 1 (top) and u = 0 (bottom). All the trajectories tend to V ∗(top) or V∗(bottom) as the time tends to inﬁnity. Speed Time Figure 2: Time plot of the dynamics with constant control u = 1 (top) and u = 0 (bottom). All the trajectories tend to V ∗(top) or V∗(bottom) as the time tends to inﬁnity. Time Time Time Time Speed Speed Speed Speed Figure 3: Representation of the speed with respect to the time. In this example, the initial speed x0 is assumed to be zero. Both pictures in the left column represent possible optimal solutions to Problem 1. Both pictures on the right cannot be possible optimal solutions of Problem 1 because of the red dashed parts. Time Time Figure 3: Representation of the speed with respect to the time. In this example, the initial speed x0 is assumed to be zero. Both pictures in the left column represent possible optimal solutions to Problem 1. Both pictures on the right cannot be possible optimal solutions of Problem 1 because of the red dashed parts. 6 V0 V0 + w0 Time V1 V1 + w1 Speed x2 Figure 4: First order variation argument. 2.3 Physical assumptions The reference trajectory (solid line), with extremal speeds Vj, 1 ≤j ≤4, is compared with the close neighbor (dashed line) with extremal speeds Vj + wj, 1 ≤j ≤4. In this example, w2 = w3 = 0. V0 V0 + w0 Time V1 V1 + w1 Speed x2 Figure 4: First order variation argument. The reference trajectory (solid line), with extremal speeds Vj, 1 ≤j ≤4, is compared with the close neighbor (dashed line) with extremal speeds Vj + wj, 1 ≤j ≤4. In this example, w2 = w3 = 0. V0 V0 + w0 V1 V1 + w1 Speed x2 Time Figure 4: First order variation argument. The reference trajectory (solid line), with extremal speeds Vj, 1 ≤j ≤4, is compared with the close neighbor (dashed line) with extremal speeds Vj + wj, 1 ≤j ≤4. In this example, w2 = w3 = 0. 2.4 Oscillating structure of the solutions of Problem 1 From (6), the invariance of total time (race duration equals T) implies , p , p From (6), the invariance of total time (race duration equals T) implies 6), the invariance of total time (race duration equals T) implies p X j=1 wj 1 f(Vj, 1) − 1 f(Vj, 0) + l X j=1 τj + l′ X j=1 τ ′ j = 0. (9) (9) From (7), the invariance of the average speed (covered length is equal to L) implies From (7), the invariance of the average speed (covered length is equal to L) implies From (7), the invariance of the average speed (covered length is equal to L) implies p X j=1 wjVj f(Vj, 1) − wjVj f(Vj, 0) + V ∗ l X j=1 τj + V∗ l′ X j=1 τ ′ j = 0. (10) (10) The optimality of u implies the invariance at ﬁrst order of the cost. Hence, from (8), The optimality of u implies the invariance at ﬁrst order of the cost. Hence, from (8), p X j=0 wjh(Vj, 1) f(Vj, 1) + h(V ∗, 1) l X j=1 τj + h(V∗, 0) l′ X j=1 τ ′ j = 0. From Assumption 2.4, p X j=0 wjh(Vj, 1) f(Vj, 1) + h(V ∗, 1) l X j=1 τj = 0, or, deﬁning ˜wj = wj 1 f(Vj,1) − 1 f(Vj,0) , Θ = Pl j=1 τj and Θ′ = Pl′ j=1 τ ′ j p X j=0 ˜wj h(Vj, 1)f(Vj, 0) f(Vj, 0) −f(Vj, 1) + h(V ∗, 1)Θ = 0. (11) p X j=0 wjh(Vj, 1) f(Vj, 1) + h(V ∗, 1) l X j=1 τj + h(V∗, 0) l′ X j=1 τ ′ j = 0. From Assumption 2.4, From Assumption 2.4, p X j=0 wjh(Vj, 1) f(Vj, 1) + h(V ∗, 1) l X j=1 τj = 0, or, deﬁning ˜wj = wj 1 f(Vj,1) − 1 f(Vj,0) , Θ = Pl j=1 τj and Θ′ = Pl′ j=1 τ ′ j p X j=0 ˜wj h(Vj, 1)f(Vj, 0) f(Vj, 0) −f(Vj, 1) + h(V ∗, 1)Θ = 0. (11) p X j=0 ˜wj h(Vj, 1)f(Vj, 0) f(Vj, 0) −f(Vj, 1) + h(V ∗, 1)Θ = 0. 2.4 Oscillating structure of the solutions of Problem 1 , TVp where u has discontinuities, successive global maxima V ∗on some intervals of length T1, T2, . . . , Tl and successive global minima V∗on some intervals of length T ′ 1, T ′ 2, . . . , T ′ l′. We insist on the fact that V1, V2, . . . , Vp belong to the closed interval [V∗, V ∗]. That is, the same control u may be associated with several diﬀerent representations. For instance, a control u that switches from 1 to 0 exactly when the speed reaches V ∗can be represented either with some TVi with Vi = V ∗, or with some Ti = 0 (meaning that the maximal speed is constant during a time equal to zero). Note also that we only consider extrema of x on the open interval (0, T) (i.e., we do not consider the initial and ﬁnal acceleration/deceleration if any). We proceed with a classical argument of calculus of variation and introduce uw,τ, an other admissible control for average speed V , close to u in norm L1, which admits local minima Vj + wj, local maxima Vj + wj and (if applicable 7 for the reference trajectory) global extrema V∗and V ∗on some intervals of length T1 + τ1, T2 + τ2, . . . , Tl + τl. If Ti = 0 or Vi = V∗, we impose τi ≥0. If Vi = V ∗, we impose wi ≤0. ( ) ( ) for the reference trajectory) global extrema V∗and V ∗on some intervals of length T1 + τ1, T2 + τ2, . . . , Tl + τl. If Ti = 0 or Vi = V∗, we impose τi ≥0. If Vi = V ∗, we impose wi ≤0. ( ) h f l ( d l ) l for the reference trajectory) global extrema V∗and V ∗on some intervals of length T1 + τ1, T2 + τ2, . . . , Tl + τl. If Ti = 0 or Vi = V∗, we impose τi ≥0. If Vi = V ∗, we impose wi ≤0. From (6), the invariance of total time (race duration equals T) implies If Ti = 0 or Vi = V∗, we impose τi ≥0. If Vi = V , we impose wi ≤0. 2.4 Oscillating structure of the solutions of Problem 1 (11) (11) Considering the left hand sides of equations (9), (10) and (11) as linear forms in ( ˜wj, Θ, Θ′), we infer that the matrix Considering the left hand sides of equations (9), (10) and (11) as linear forms in ( ˜wj, Θ, Θ′), we infer that the matrix   1 . . . 1 1 1 V1 . . . Vp V ∗ V∗ h(V1,1)f(V1,0) f(V1,0)−f(V1,1) . . . h(Vp,1)f(Vp,0) f(Vp,0)−f(Vp,1) h(V ∗, 1) 0   has rank at most two (the last two columns only appears when l ̸= 0 or l′ ̸= 0). Finally, notice that has rank at most two (the last two columns only appears when l ̸= 0 or l′ ̸= 0). Finally, notice that h(V ∗, 1) = h(V ∗, 1)f(V ∗, 0) f(V ∗, 0) −f(V ∗, 1) h(V∗, 0) = h(V∗, 1)f(V∗, 0) f(V∗, 0) −f(V∗, 1) = 0 h(V ∗, 1) = h(V ∗, 1)f(V ∗, 0) f(V ∗, 0) −f(V ∗, 1) and h(V∗, 0) = h(V∗, 1)f(V∗, 0) f(V∗, 0) −f(V∗, 1) = 0 and and to complete the proof of Lemma 2. to complete the proof of Lemma 2. Proof of Proposition 1: We proceed by contradiction and assume that the speed admits p diﬀerent local extrema. If p = 2, there is nothing to prove. Else, we consider the ﬁrst three columns of the matrix obtained in Lemma 2. Up to permutation of the indices, we may assume without loss of generality that V1 < V3 < V2. Find µ, ν > 0, µ + ν = 1 such that V3 = µV1 + νV2. Since the matrix   1 1 1 = µ.1 + ν.1 V1 V2 V3 = µV1 + νV2 h(V1,1)f(V1,0) f(V1,0)−f(V1,1) h(V2,1)f(V2,0) f(V2,0)−f(V2,1) h(V3,1)f(V3,0) f(V3,0)−f(V3,1)   has rank at most 2, its determinant is zero. Hence, h(V3, 1)f(V3, 0) f(V3, 0) −f(V3, 1) = µ h(V1, 1)f(V1, 0) f(V1, 0) −f(V1, 1) + ν h(V2, 1)f(V2, 0) f(V2, 0) −f(V2, 1). Hence, the mapping F : x 7→ h(x,1)f(x,0) f(x,1)−f(x,0) is not strictly convex or concave, which is in contradiction with A ti 2 7 ■ Hence, the mapping F : x 7→ h(x,1)f(x,0) f(x,1)−f(x,0) is not strictly convex or concave, which is in contradiction with Assumption 2.7. 3.2 Main Result The main result, proved below, stipulates that the minimum of Ca is reached by controls with two disconti- nuities. In other words, the most energy eﬃcient driving strategy is to let the speed x periodically oscillate between two values V a and V b, with no intermediate local extremum. Precisely, one has the following result (recall that V ∗and V∗are deﬁned in Assumption 2.1): Proposition 3 Under Assumptions 1 and 2, for every closed interval I contained in [V∗, V ∗] and every interior point V of I, Ca admits a minimum on UI V , and inf UI V Ca= min U2,I V Ca. The result of Proposition 3 is twofold: ﬁrst, there exists a minimum of Ca and second, this minimum is reached with periodic controls with two discontinuities for a period. The second part is a direct application of Proposition 1. The only point to check is the existence of solutions, which is done in Section 3.3. roblem 2 Let V > 0 be given. Find the minimum, if any, of Ca on UI V . Problem 2 Let V > 0 be given. Find the minimum, if any, of Ca on UI V . Problem 2 Let V > 0 be given. Find the minimum, if any, of Ca on UI V . Problem 2 may be seen as an inﬁnite time horizon version of Problem 1. Indeed, no ﬁnal time is given. We only aim at ﬁnding the lowest possible energy consumption for a long distance run at average speed V . 2.4 Oscillating structure of the solutions of Problem 1 ■ g F : x 7→ h(x,1)f(x,0) f(x,1)−f(x,0) is not strictly convex or concave, which is in contradiction with ■ 8 3.3 Proof The proof requires several intermediate results and is given in the following section. 3.1 Statement of the optimal problem In this subsection, we consider Problem 1 with a control restricted to a periodic control. Precisely, for any given V > 0, we deﬁne the admissible trajectories with average speed V as the curves t ∈[0, T1] 7→(x(t), u(t)) ∈ R × {0, 1} for some suitable T1 > 0, with x absolutely continuous and u piecewise constant, satisfying (1) and such that x(0) = x(T1) = V , u(0) = u(T1) and 1 T1 R T1 0 x(t)dt = V . In particular, the control u has an even number of discontinuities in [0, T1]. In this subsection, we consider Problem 1 with a control restricted to a periodic control given V > 0, we deﬁne the admissible trajectories with average speed V as the curves t ∈[0, [ ] A piecewise constant function u : [0, T1] →{0, 1} is an admissible control for average speed V if there exists a function x : [0, T1] →R such that (x, u) is an admissible trajectory with average speed V . For T1 ≥0 and k in N, we denote with Uk V (T1) the set of admissible controls for average speed V deﬁned on [0, T1] with at most k discontinuities, and we deﬁne the sets UV (T1) = ∪k∈NUk V (T1), Uk V = ∪T1>0Uk V (T1) and UV = ∪T1>0UV (T1) = ∪k∈NUk V . For any subset I of R, we denote respectively with Uk,I V (T1), UI V (T1) and UI V the functions of Uk V (T1), UV (T1) and UV where x takes value in I. ) V inally, we associate, with every admissible control for average speed V in UV (T1), the cost Ca(u) = 1 T1 C(u) which can be seen as the average energy consumption to run the distance V T1 in time T1. Let us consider the following problem. which can be seen as the average energy consumption to run the distance V T1 in time T1. Let us consider the following problem. 3.3.2 Cost estimates for control with small periods Lemma 5 For every k in 2N, limT1→0 mk V (T1) = +∞ Proof: We ﬁrst prove by contradiction that Uk V (T1) does not contain any continuous function. Indeed, assume that u in Uk V (T1) is continuous. Then, the associated trajectory x is decreasing (if u = 0) or increasing (if u = 1). Since u belongs to Uk V (T1), x(0) = x(T1). Hence x is constant. By Assumptions 2.1 and 2.3, x = V ∗ or x = V∗. Hence, the average of x is not V (which belongs to (V∗, V ∗)). This is a contradiction with the fact that u belongs to Uk V (T1). Hence u has at least two discontinuities and N(u) ≥1. Thus, Ca(u) > α/T1. ■ Remark 2 As α tends to zero, the interval between two consecutive switching times tends to zero. Hence, the velocity tends to a constant V , which is the optimal trajectory of the relaxed problem when u can take value in [0, 1] with f(x1, x2, u) = uf(x1, x2, 1) + (1 −u)f(x1, x2, 0). 3.3.3 Oscillating structures of optimal trajectories with a given period Lemma 6 For every k in 2N, for every positive Ta, there exists p in N such that mk V (Ta) = m2 V (Ta/p) Lemma 6 For every k in 2N, for every positive Ta, there exists p in N such that mk V (Ta) = m2 V (Ta/p) Proof: This is nothing but a restatement of Proposition 1 in the periodic case. ■ Proof: This is nothing but a restatement of Proposition 1 in the periodic case. 3.3.1 Existence of solutions to Problem 2 for a given number of switches 3.3.1 Existence of solutions to Problem 2 for a given number of switches Lemma 4 For every even k in N and T1 > 0, Ca (and hence C) admits a minimum mk V (T1) on Uk V (T1). Lemma 4 For every even k in N and T1 > 0, Ca (and hence C) admits a minimum mk V (T1) on Uk V (T1). Lemma 4 For every even k in N and T1 > 0, Ca (and hence C) admits a minimum mk V (T1) on Uk V (T1). Proof: This is a consequence of the compactness of Uk V (T1) endowed with the L1 norm and of the continuity of C when the number of switches is constant equal to k. ■ 9 Time T2 T1 Speed Speed Time T2 T1 Tm V∗ V ∗ V0 V1 Td Time T2 T1 Speed Speed Time T2 T1 Tm V∗ V ∗ V0 V1 Td Figure 5: Large period optimal controls. For a control in U2 V (T2), we deﬁne T1 as the acceleration duration in the case where trajectories are backward unique near V1 (top) and as the time where x = V ∗when backward uniqueness does not hold (bottom). The ratio T1/T2 is the same at ﬁrst order in both cases. Figure 5: Large period optimal controls. For a control in U2 V (T2), we deﬁne T1 as the acceleration duration in the case where trajectories are backward unique near V1 (top) and as the time where x = V ∗when backward uniqueness does not hold (bottom). The ratio T1/T2 is the same at ﬁrst order in both cases. 3.3.4 Cost estimates for controls with large periods 5 bottom) Tm = R V ∗ V∗ ds f(s,1) and Td = R V ∗ V∗ ds f(s,0). Hence, In the case where the trajectories of (1) reach V ∗and V∗in ﬁnite time, we denote (see Fig. 5 bottom) Tm = R V ∗ V∗ ds f(s,1) and Td = R V ∗ V∗ ds f(s,0). Hence, T1 T2 (V ∗−V∗) = V −V∗+ (Tm + Td) V T2 (15) (15) Plugging (15) in the deﬁnition of Ca, we ﬁnd that the estimate (14) is still valid (with ϵ = 0) when the trajectories of (1) are not backward uniquely deﬁned near V ∗and V∗. The mixed case (the trajectories are locally backward unique near V ∗and reach V∗in ﬁnite time, or vice versa) can be treated as the two cases above and is omitted. Plugging (15) in the deﬁnition of Ca, we ﬁnd that the estimate (14) is still valid (with ϵ = 0) when the trajectories of (1) are not backward uniquely deﬁned near V ∗and V∗. The mixed case (the trajectories are locally backward unique near V ∗and reach V∗in ﬁnite time, or vice versa) can be treated as the two cases above and is omitted. In conclusion, the estimate (14) is valid in any case and Lemma 7 follows from Assumption 2.6. ■ From Lemmas 5 and 7, we deduce that the continuous mapping T 7→mk V (T) admits a minimum on (0, +∞). We denote this quantity with mk V := minT1>0 mk V (T1). 3.3.4 Cost estimates for controls with large periods From (4), we get that Z V1 s Z V0 s T2V = Z V1 V0 s f(s, 1)ds + Z V0 V1 s f(s, 0)ds H T2V = Z V1 V0 s f(s, 1)ds + Z V0 V1 s f(s, 0)ds T2V = Z V1 V0 s f(s, 1)ds + Z V0 V1 s f(s, 0)ds Hence, V ∗T1 T + V∗ 1 −T1 T + 1 T Z V1 s −V ∗ f(s 1) ds Z V0 f( , ) Z V1 f( , ) Hence, V ∗T1 T2 + V∗ 1 −T1 T2 + 1 T2 Z V1 V0 s −V ∗ f(s, 1) ds + 1 T Z V0 s −V∗ f( 0)ds = V (12) Hence, V ∗T1 T2 + V∗ 1 −T1 T2 + 1 T2 Z V1 V0 s −V ∗ f(s, 1) ds + 1 T2 Z V0 V1 s −V∗ f(s, 0)ds = V (12) (12) that implies limT2→∞T1 = +∞. Hence, limT2→∞V0 = V∗and limT2→∞V1 = V ∗. Thus, there exists a function ϵ such that lim+∞ϵ = 0 and that implies limT2→∞T1 = +∞. Hence, limT2→∞V0 = V∗and limT2→∞V1 = V ∗. Thus, there exists a function ϵ such that lim+∞ϵ = 0 and T1 T2 = − 1 T2(V ∗−V∗) Z V ∗ V∗ s −V ∗ f(s, 1) ds + Z V∗ V ∗ s −V∗ f(s, 0)ds ! + V −V∗ V ∗−V∗ + ϵ(T2) T2 . (13) (13) We infer from (3) and (13) that We infer from (3) and (13) that We infer from (3) and (13) that Ca = α T2 + 1 T2 Z V1 V0 h(s, 1) f(s, 1)ds = α T2 + 1 T2 Z V1 V0 h(s, 1) −h(V ∗, 1) f(s, 1) ds + h(V ∗, 1)T1 ! = h(V ∗, 1) V −V∗ V ∗−V∗ + 1 T2 " α + Z V ∗ V∗ h(s, 1) −h(V ∗, 1) f(s, 1) ds − Z V ∗ V∗ s −V ∗ f(s, 1) ds + Z V∗ V ∗ s −V∗ f(s, 0)ds ! h(V ∗, 1) V ∗−V∗ # +ϵ(T2) T2 +ϵ(T2) T2 (14) +ϵ(T2) T2 (14) (14) In the case where the trajectories of (1) reach V ∗and V∗in ﬁnite time, we denote (see Fig. 3.3.4 Cost estimates for controls with large periods Lemma 7 For every T1 large enough, m2 V (T1) ≤lim infT2→+∞m2 V (T2). Proof: We do the proof by computing an expansion of m2 V (T2) as T2 tends to inﬁnity. L u a minimizing control in m2 V (T2). We distinguish among several cases, depending on the Proof: We do the proof by computing an expansion of mV (T2) as T2 tends to inﬁnity. Let T2 be given, and u a minimizing control in m2 V (T2). We distinguish among several cases, depending on the regularity of f. In the case of local backward uniqueness of the trajectories of (1) in neighborhoods of V ∗and V∗(see Fig. 5 top), we may assume, up to a time shift, that u(t) = 0 for t ∈(0, T1) and u(t) = 0 for t ∈(T1, T2). The associated trajectory reaches its minimum V0 at t = 0 and its maximum V1 at T1. From (3), we get that In the case of local backward uniqueness of the trajectories of (1) in neighborhoods of V ∗and V∗(see Fig. 5 top), we may assume, up to a time shift, that u(t) = 0 for t ∈(0, T1) and u(t) = 0 for t ∈(T1, T2). The associated trajectory reaches its minimum V0 at t = 0 and its maximum V1 at T1. From (3), we get that Z V1 V0 s f(s, 1)ds = Z V1 V0 s −V ∗ f(s, 1) ds + V ∗T1 and 10 Z V0 V1 s f(s, 0)ds = Z V0 V1 s −V∗ f(s, 0)ds + V∗(T2 −T1). From (4), we get that T2V = Z V1 V0 s f(s, 1)ds + Z V0 V1 s f(s, 0)ds Hence, Z V0 V1 s f(s, 0)ds = Z V0 V1 s −V∗ f(s, 0)ds + V∗(T2 −T1). Z V0 V1 s f(s, 0)ds = Z V0 V1 s −V∗ f(s, 0)ds + V∗(T2 −T1). 4.1 Modeling For a real vehicle, the autonomous dynamics described in Section 2 is too restrictive. Various types of forces act on the vehicle. Some of them, like the solid friction forces applied on the wheels or axes, are roughly constant. Some others, such as the gravitation, depends on the slope of track. Some of the forces also depend on many (partly unknown) external factors, including the actual weather. This is the case for the aerodynamic drag depending on the velocity, the orientation of the track with respect to the wind and the strength of the wind. If we consider both the position and the velocity of the vehicle denoted respectively x1 and x2, the dynamics turns into: ˙ ˙x1 = x2 ˙x2 = f(x1, x2, t, u) (16) (16) with f : [0, L] × [0, +∞) × [0, T] × {0, 1} →R a continuous function. The control u : [0, T] →{0, 1} is a piecewise constant function accounting for the state of the engine (oﬀwhen u = 0, on when u = 1). with f : [0, L] × [0, +∞) × [0, T] × {0, 1} →R a continuous function. The control u : [0, T] →{0, 1} is a piecewise constant function accounting for the state of the engine (oﬀwhen u = 0, on when u = 1). The steering possibilities of the car being limited at high speed, we introduce a (smooth) function x1 7→ V s(x1) representing the maximal safety speed at point x1 and we will only consider solutions of system (16) satisfying the state constraint 0 ≤x2(t) ≤V s(x1(t)) for every t. y g ( ) ( ( )) y As in Section 2.1, with every admissible control u and absolutely continuous solution (x1, x2) of (16), we associate the energy cost C(u) = Z T 0 h(x2(t), u(t))dt + α N(u) (17) (17) where h : [0, +∞)×{0, 1} →[0, +∞) is a C1 function, α is a positive real constant and N denotes the number of discontinuities of u. where h : [0, +∞)×{0, 1} →[0, +∞) is a C1 function, α is a positive real constant and N denotes the number of discontinuities of u. 3.4 Practical determination of the upper and lower speed limits Based on the knowledge of the average target speed V , the dynamics f and the cost function h, the search for the upper and lower speeds of the optimal solution whose existence is asserted by Proposition 3 amounts to solve two 1D continuous optimization problems. A variety of methods are available in the literature, especially when some regularity assumptions are done on f and h. We present below a naive yet eﬃcient grid method. Step 1: candidate selection Select p candidates V a 1 < V a 2 < . . . < V a p < V for the lower speed limit. Step 2: search for upper limit For every i = 1 . . . p, ﬁnd (by dichotomy), the upper limit V b i ensuring that the average speed is V (in the case where V b i = V ∗, compute also the time where the velocity is constant). Step 3: cost for each candidate For every i = 1 . . . p, compute the cost Ca associated with a trajectory oscillating from V a i to V b i (or staying at V ∗for the time computed in Step 2). p p ( i , i ) In our implementation (see Section 5), we mainly used p = 4, V a i = V −5−i 2 m.s−1, i = 1..4. 4.1 Modeling Problem 3 We aim to ﬁnd a piecewise constant control u that steers (x1, x2), subject to the velocity con- straints x2(t) ∈V s(x1(t)) for every t and the dynamic (16), from (0, 0) at time t = 0 to (L, x2(T)) in time less than T and minimizes cost C (17). We do the following assumptions. Assumption 3 1. The track and weather conditions are piecewise continuous: there exist two subdivisions 0 = x0 1 < x1 1 < . . . < xp1 1 = L and 0 = t0 < t1 < . . . < tpt = T of [0, L] and [0, T] such that for every u in {0, 1}, (x1, x2, t) 7→f(x1, x2, t, u) is continuous on [xj 1, xj+1 1 ) × R × [tl, tl+1) for every j < p1 and l < pt; Assumption 3 1. The track and weather conditions are piecewise continuous: there exist two subdivisions 0 = x0 1 < x1 1 < . . . < xp1 1 = L and 0 = t0 < t1 < . . . < tpt = T of [0, L] and [0, T] such that for every u in {0, 1}, (x1, x2, t) 7→f(x1, x2, t, u) is continuous on [xj 1, xj+1 1 ) × R × [tl, tl+1) for every j < p1 and l < pt; 2. For u constant in {0, 1}, for every j < p1 and l < pt, for every (x1(t0), x2(t0), t0) in [xj 1, xj+1 1 ) × R × [tl, tl+1), the Cauchy problem (16) with initial condition (x1(t0), x2(t0), t0) at time t0 admits a locally unique solution in positive time; 2. For u constant in {0, 1}, for every j < p1 and l < pt, for every (x1(t0), x2(t0), t0) in [xj 1, xj+1 1 ) × R × [tl, tl+1), the Cauchy problem (16) with initial condition (x1(t0), x2(t0), t0) at time t0 admits a locally unique solution in positive time; 3. For every time t∗in [0, T], for every position on the track x∗ 1 in [0, L], the functions (x, u) 7→f(x∗ 1, x, t∗, u) and (v, u) 7→h(v, u) satisfy Assumptions 1 and 2. 3. 3.3.5 Completion of the proof of Proposition 3 By deﬁnition, infUV Ca = infT >0 infk∈2N mk V (T). By Lemma 4, each optimization problem for a given pe- riod T1 admits a solution, and by Lemma 6, the associated control can be chosen with two discontinuities infUV Ca = infT >0 m2 V (T), this last quantity being equal to m2 V by Lemmas 5 and 7. In particular, the inﬁmum infUV Ca is reached and is indeed a minimum. ■ 11 4.3 Well-posedness The only point to check to guarantee the well-posedness of the control scheme S is that the interval between two consecutive engine switches cannot tend to zero. Indeed, let ut be an admissible optimal control for average speed V computed at time t. We denote the period of ut with Tt and the associated cost with Ca t (index t is the time at which the control has been computed). By Lemma 5, Ca(ut) > α/T1, hence Tt > α/Ca(ut). The cost t 7→Ca t is continuous and the interval [0, T] is compact, hence inf0≤t≤T Ca t > 0. And thus, inft∈[0,T ] Tt > 0, and the switching points cannot accumulate (no Zeno phenomena). 4.2 Description of the strategy The control strategy S is to repeat the following until t = (n + 1)ta: The control strategy S is to repeat the following until t = (n + 1)ta: 1. Keep u = 1 until x2(t) reaches V b(nta). 2. As soon as x2(t) reaches V b(nta), set u = 0. 3. Keep u = 0 until x2(t) = V a(t), then switch to u = 1. 4. Come back to (1), unless x1(t) ≥L. 4.1 Modeling For every time t∗in [0, T], for every position on the track x∗ 1 in [0, L], the functions (x, u) 7→f(x∗ 1, x, t∗, u) and (v, u) 7→h(v, u) satisfy Assumptions 1 and 2. 12 Assumption 3.3 means that, around x∗ 1 in [0, L], we can locally replace the actual vehicle evolving on the actual irregular track with varying weather conditions by a ﬁctional vehicle evolving on an inﬁnite ﬁctional track with constant slope or wind velocity (at x∗ 1). p y ( 1) Let u : [0, T1] →{0, 1} be piecewise constant. A pair (x1, x2) : [0, T1] →R2 is a solution of (16) if (x1, x2) is continuous, piecewise diﬀerentiable and satisﬁes (16) for almost every time t in [0, T1]. As before, the existence (but not the uniqueness) of such solutions to Cauchy problem (16) with initial condition (x1(t0), x2(t0)) at time t0 is a consequence of Assumption 3.1 and Peano theorem (see for instance [12], Theorem 2.1 page 10). 4.2 Description of the strategy The strategy we propose can be split in two parts: ﬁrst, we identify the current dynamics and compute the current target and then we use the results of Section 2 during a small time interval (typically three seconds in our case), after which we proceed to a new identiﬁcation, and a new optimization. Precisely, we split the time interval [0, T] into small intervals [0, T] = SN n=0[nta, (n + 1)ta]. Assume that current time is equal to t = nta for some n, and the vehicle is at position x1(nta) with velocity x2(nta). Identify the current dynamics (16) with x∗ 1 = x1(nta) and t∗= nta and compute the target average speed V = L−x2(nta) T −nta . If V < V s(x1(ta)), ﬁnd the minimum m2 V and the corresponding trajectory (x2, u∗) of (1), with the constraint x2 ≤V s(x1(ta)). This can be done, for instance, with Step 1 to Step 4 detailed in Section 3.4. The optimal speed x2 oscillates between two values V a(ta) and V b(ta). Assume that current time is equal to t = nta for some n, and the vehicle is at position x1(nta) with velocity x2(nta). Identify the current dynamics (16) with x∗ 1 = x1(nta) and t∗= nta and compute the target average speed V = L−x2(nta) T −nta . If V < V s(x1(ta)), ﬁnd the minimum m2 V and the corresponding trajectory (x2, u∗) of (1), with the constraint x2 ≤V s(x1(ta)). This can be done, for instance, with Step 1 to Step 4 detailed in Section 3.4. The optimal speed x2 oscillates between two values V a(ta) and V b(ta). p p ( ) ( ) If V b(ta) > V s(x1(ta), keep V b(ta) = V s(x1(ta)) and V a(ta) = V s(x1(ta)) −δ where δ is a small enough constant (for our application, δ = 0.5 m.s−1). p p ( ) ( ) If V b(ta) > V s(x1(ta), keep V b(ta) = V s(x1(ta)) and V a(ta) = V s(x1(ta)) −δ where δ is a small enough constant (for our application, δ = 0.5 m.s−1). ( ) If V b(ta) > V s(x1(ta), keep V b(ta) = V s(x1(ta)) and V a(ta) = V s(x1(ta)) − constant (for our application, δ = 0.5 m.s−1). 4.4 Constraints fulﬁllment and robustness The proposed strategy happens to be extremely robust in practice. This robustness may be explained by two reasons of diﬀerent natures. The ﬁrst reason for the robustness of the strategy is the continuous actualization of the target average speed. The second reason for robustness is the fact that we use a control on the average of the velocity. We ﬁrst state two abstract results (Lemma 8 and Proposition 9 below). Lemma 8 Let Va < Vb be given, denote with C the set of not vanishing continuous function on [Va, Vb]. For g in C, denote T(g) = Z Vb Va ds g(s), L(g) = Z Vb Va sds g(s) and the mapping F deﬁned on C by F : g 7→L(g)/T(g) ∈R. δ in C and every continuous δg such that ∥δg g ∥L∞< 1, . Then, for every g in C and every continuous δg such that ∥δg g ∥L∞< 1, F(g + δg) −F(g) = 1 T(g + δg) ∞ X n=1 (−1)n Z Vb Va s −F(g) g(s) δg(s) g(s) n ds. (18) (18) 13 Constant Value Comment a 6.10−4 m−1 identiﬁed c 3.10−2 m.s−2 identiﬁed g 9.81 m.s−2 tabulated f1 0.20 m.s−2 tabulated m 93 kg measured Figure 6: Numerical value of the constants appearing in the dynamics (19). Figure 6: Numerical value of the constants appearing in the dynamics (19). Figure 6: Numerical value of the constants appearing in the dynamics (19). Proof: Standard expansions show that L(g + δg) = X n≥0 (−1)n Z Vb Va s(δg)n gn+1(s)ds, and T(g + δg) = X n≥0 (−1)n Z Vb Va (δg)n gn+1(s)ds, for δg small enough. Conclusion follows with basic calculus. ■ and T(g + δg) = X n≥0 (−1)n Z Vb Va (δg)n gn+1(s)ds, for δg small enough. Conclusion follows with basic calculus. ■ Proposition 9 If δg/g is constant, then F(g + δg) = F(g). Proposition 9 If δg/g is constant, then F(g + δg) = F(g). Proposition 9 If δg/g is constant, then F(g + δg) = F(g). t Z Vb Va s −F(g) g(s) ds = L(g) −F(g)T(g) = 0. ■ f L 8 d P i i 9 If f i ﬁ A i 1 d 2 d a Let us comment the statements of Lemma 8 and Proposition 9. 5 Eﬀective implementation The strategy proposed in Section 4.2 has been implemented in an oﬃcial competition in 2 The strategy proposed in Section 4.2 has been implemented in an oﬃcial competition in 2014. 4.4 Constraints fulﬁllment and robustness If f satisﬁes Assumptions 1 and 2 and one choses g : s 7→f(s, 1), the number F(g) is the average speeds when one speed up from Va to Vb with the dynamics f. If the dynamics s 7→f(s, 1) has been incorrectly identiﬁed and so is ˜g : s 7→f(s, 1) + δf(s), Lemma 8 gives an estimate of the diﬀerence between the expected average speed F(˜g) and the actual average speed F(g). The point of Proposition 9 is that, if (δf(s))/f(s, 1) is constant, then the change of average speed is zero. Hence the diﬀerence F(g + δg) −F(g) is due to the variance of δg g which is usually small with respect to δg g . The same computation is obviously valid for decelerations if one considers g : s 7→f(s, 0), and generically in the non-autonomous case if one considers g : s 7→f(x1(s), s, t(s), u) where x1(s) and t(s) are respectively the position on the track and the time where the vehicle reaches speed s. Proof: Standard expansions show that 6 Conclusions A real-time adaptive driving command strategy for low consumption vehicle has been presented. Real life implementation on a competition prototype showed good agreement between the theoretical analysis and the eﬀective results. The consumption performances are comparable with performances obtained by highly trained human drivers. All the computations are done on board, in real time, with a low cost micro controller and limited energy consumption. gy Among the various advantages of the automatic command, the pilots underlined the dramatic safety im- provement (no need to concentrate on the velocity anymore) and the robustness of the command that allows eﬀective recovering of optimal timing after traﬃc perturbations. From a more theoretical point of view, it has been provided an explicit hybrid control strategy in a form of a piecewise constant control (on/oﬀ) for low regular dynamics, in particular not Lipschitz continuous. Furthermore, a switching cost has been considered. Because of the heavy computational burden, the use of a classical PMP is redhibitory. This contribution acts an an eﬃcient alternative from this perspective. Optimality of the control in the case of autonomous systems and the robustness in the case of non autonomous systems (with time-varying disturbances) are shown. 5.1 Vir’Volt 2 prototype Figure 7: Comparisons of the oﬃcial runs results for the EMT prototype at Shell Eco-Marathon 2013 (manual command) and 2014 (automatic command and strategy of Section 4.2). of the power conversion block. In the following, we consider the case where the power consumption is roughly independent from speed, h2(x2, 1) = 23 · 7 = 161 W for x2 > 5 m.s−1. Notice that the dynamics f completed with h1 or h2 satisﬁes in both cases Assumption 3 at any time. of the power conversion block. In the following, we consider the case where the power consumption is roughly independent from speed, h2(x2, 1) = 23 · 7 = 161 W for x2 > 5 m.s−1. Notice that the dynamics f completed with h1 or h2 satisﬁes in both cases Assumption 3 at any time. For a ﬂat horizontal track without wind, with α = 10 J and a target average speed of 7 m/s, the optimal strategy consists in letting the speed oscillate between 6.1 m/s and 7.94 m/s. The corresponding energetic cost for a track of 16.5 km (similar to the editions 2013 and 2014 of the European Shell Eco-Marathon) is 104 189 J. This computation is given for reference only since it neglects the fact that the vehicle start from velocity zero (kinetic energy of the vehicle at 7 m/s: about 2.5 kJ), the inﬂuence of the wind, and the elevation of the actual track which is not perfectly horizontal. 5.2 Eﬀective results Computation of the control law (including identiﬁcation of the current dynamics from the sensors data) is possible with an average power of 10 mW. The oﬃcial results of the EMT prototype measured in the 2014 edition of the Shell Eco-Marathon are presented in Figure 7 with the ﬁrst result of the 2013 edition. The 2013 prototype is basically the same as the 2014 one without embedded control (the driver directly switches the engine on or oﬀ). These results are directly comparable since they all were obtained without solar cells (EMT record on the Rotterdam track is 90211 J and was obtained with manual velocity command during a particularly sunny test in May 2013). 5.1 Vir’Volt 2 prototype The prototype (including the pilot) has a total weight of about 90 kg, is driven by a 200 W electric DC motor powered by a 23 V battery. The control scheme presented in this note is implemented on a microcontroller (ref dsPIC33ep512mu810 from MicrochipTM), with a 140 MHz internal clock. Standard 32 kHz oscillators, bike velocity sensors and GPS receiver provide (respectively) the current time, the velocity and the position needed for the algorithm. A simpliﬁed dynamics for Vir’Volt 2 is given by A simpliﬁed dynamics for Vir’Volt 2 is given by f(x1, x2, t, u) = −a(x2−v(x1, t))2 −c sign(x2) −g sin θx1 + f1 if u = 1 0 if u = 0 (19) (19) where v(x1, t) is the speed of the wind at point x1 at time t, θx1 is the angle of the track with the horizontal plane at point x1, f1 is the traction force by mass unit when the engine is on, g is the gravitational acceleration, a accounts for the aerodynamics drag, and c accounts for the solid friction in the vehicle. where v(x1, t) is the speed of the wind at point x1 at time t, θx1 is the angle of the track with the horizontal plane at point x1, f1 is the traction force by mass unit when the engine is on, g is the gravitational acceleration, a accounts for the aerodynamics drag, and c accounts for the solid friction in the vehicle. The traction force f1 is tabulated from the engine data sheet. There are several ways to consider the energetic cost. The power transmitted to the wheels is the product h1(x2, 1) = x2 · m · f1. The energy taken from the battery is always larger than h1 and depends on the design 14 2013 2014 1st run 1st run 2nd run Run duration expected 35m 00s 36m 00s 38m 30s realized 36m 01s 36m 10s 38m 31s Consumption 112 742 J 120 150 J 107 946 J Figure 7: Comparisons of the oﬃcial runs results for the EMT prototype at Shell Eco-Marathon 2013 (manual command) and 2014 (automatic command and strategy of Section 4.2). Figure 7: Comparisons of the oﬃcial runs results for the EMT prototype at Shell Eco-Marathon 2013 (manual command) and 2014 (automatic command and strategy of Section 4.2). Ackowledgements This work has been ﬁnancially supported by CPER MISN and the F´ed´eration Charles Hermite. It is a pleasure for the authors to thank the “Eco Motion Team” in charge of the Vir’Volt prototype development, especially 15 G´erard D´echenaud, Jean-Claude Sivault, Mathieu Rupp and Sandra Rossi. References [1] G. 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https://openalex.org/W2074419882	https://scielo.conicyt.cl/pdf/rchnat/v79n1/art01.pdf, http://www.scielo.cl/pdf/rchnat/v79n1/art01.pdf, https://www.redalyc.org/pdf/3699/369944277001.pdf	es		Nudibranchia from the remote southern Chilean Guamblin and Ipún islands (Chonos Archipelago, 44-45° S), with re-description of Rostanga pulchra MacFarland, 1905	Revista chilena de historia natural	2,006	cc-by	4,930	NUDIBRANCHIA FROM SOUTHERN CHILE Revista Chilena de Historia Natural 3 79: 3-12, 2006 Nudibranchia from the remote southern Chilean Guamblin and Ipún islands (Chonos Archipelago, 44-45° S), with re-description of Rostanga pulchra MacFarland, 1905 Nudibranquios de las islas Guamblin e Ipún (Archipiélago de los Chonos 44-45° S) con redescripción de Rostanga pulchra MacFarland, 1905 MICHAEL SCHRÖDL 1 & JOSÉ H GRAU2 1 Zoologische Staatssammlung München, Münchhausenstrasen 21, 81247 München, Germany; e-mail: schroedl@zi.biologie.uni-muenchen.de 2 Instituto de Zoología, Universidad Austral de Chile, Valdivia, Chile; e-mail: androctonus@hotmail.com ABSTRACT The southern Chilean archipelago and fjordland (41-52° S) thus far is very poorly investigated also with regard to its nudibranch fauna. This study presents the first records of nudibranchs from remote islands of the Chonos archipelago exposed to the open Pacific. Collecting data and some biological observations are given for the doridoidean Archidoris fontaini, Diaulula punctuolata and Rostanga pulchra. Taxonomically relevant external and radular characters of R. pulchra are redescribed and compared with literature data on conspecifics from Argentina and California. The bipolar, cold-water adapted R. pulchra is critically compared with the western Atlantic, warm-water adapted R. byga. New distributional data indicates that the Chiloé Island area (41-43° S) does not represent a strict southern distributional barrier for warm-temperate eastern Pacific nudibranch species but is at least partly due to collecting bias. Key words: Nudibranchia, southern Chile, zoogeography, taxonomy, Rostanga pulchra. RESUMEN Los archipiélagos y fiordos del sur de Chile han sido muy poco estudiados y particularmente la fauna de nudibranquios. En este estudio se presentan los primeros registros de nudibranquios para islas del Archipiélago de los Chonos abiertas al Pacífico. Se indican datos de recolecta y algunas observaciones biológicas para los doridáceos Archidoris fontaini, Diaulula punctuolata y Rostanga pulchra. La morfología externa y caracteres radulares de R. pulchra son redescritos y comparados con literatura referente a ejemplares de Argentina y California. Se comparan críticamente R. byga de las aguas cálidas del Atlántico occidental con R. pulchra , especie bipolar adaptada a aguas frías. La nueva información distribucional indica que la isla de Chiloé no representa una barrera distribucional estricta para las especies de nudibranquios del Pacífico sudeste, y es en su mayor parte debido a un vacío de recolección. Palabras clave: Nudibranchia, Chile sur, zoogeografía, taxonomía, Rostanga pulchra. INTRODUCTION Chilean and Magellanic nudipleuran gastropods were monographically revised recently by Schrödl (2003). On this taxonomic basis, further systematic and zoogeographical analyses shall be undertaken. Earlier studies suggested a faunal change between cold-adapted Magellanic and warm-temperate “Peruvian” species at the northern end of Chiloé Island (41-42° S) (Marcus 1959, Brattström & Johannsen 1983). However, most nudipleuran species with a Magellanic distribution were shown to be widespread and extending more or less far to the north into central or even northern Chilean waters (Schrödl 1997a, 2003). In contrast, amphi-South American species such as the bipolar Rostanga pulchra MacFarland, 1905, as well as several nudipleuran species with a Peruvian distribution had their southernmost known geographical ranges at 41-43° S in the Pacific (see Schrödl 2003); this “Chiloé Island 4 SCHRÖDL & GRAU area” thus may either represent a true distributional barrier, or actual distributional limits may be due to collecting artifacts (Schrödl 2003). In fact, there is a general lack of distributional data regarding remote areas of the southern Chilean archipelago and continental fjordland (41-52° S), that thus far is amongst the most poorly sampled coastal areas of the world. Within the framework of exploring opisthobranchs of the southern Chilean fjord region, a catalog of 12 nudipleuran species from the protected Comau fjord system has been published recently (Schrödl et al. in press). The present study gives new information on a small collection of nudibranchs from the remote Guamblin and Ipún islands which are exposed to the open Pacific. Since the identity of Chilean Rostanga pulchra specimens has been doubted by Muniaín & Valdés (2000), R. pulchra is redescribed in more detail from a specimen from Ipún Island. The taxonomic status of the apparently bipolar R. pulchra and the similar, western Atlantic R. byga Marcus, 1958 is reevaluated critically. MATERIAL AND METHODS During a bioinventory survey at Guamblin and Ipún islands, Chonos Archipelago (Fig. 1), in January and February 2004, the junior author (JG) documented and collected nudibranchs from tide pools and the lower intertidal. Specimens were fixed in ethanol (70 %) for further taxonomic analysis. In laboratory, at least one specimen of each species from each locality was dissected under a stereomicroscope to confirm external identification. The single specimen of Rostanga pulchra was examined and redescribed in anatomical detail. Cuticular structures such as jaws and radula were analyzed with a scanning electron microscope. Reference specimens were deposited at the Zoologische Staatsammlung München (ZSM). Fig. 1: Map of the southern Chilean archipelago with the collecting sites Ipún and Guamblin islands. Mapa del archipiélago del sur de Chile mostrando los sitios de colecta, islas Guamblin e Ipún. NUDIBRANCHIA FROM SOUTHERN CHILE Dorididae s.l., Archidoris Bergh, 1878: Archidoris fontaini (D’Orbigny, 1837) (Fig. 2A and 2B) Material examined. One specimen (ZSM Moll 20040982; dissected), Guamblin Island (44° 46.6’ S, 75° 09.8’ W); tide pool, on rock. Two specimens (ZSM Moll 20040983; one dissected), Ipún Island (44° 33.1’ S, 74° 48.0’ W); tide pool, on rock. Distribution. Independence Bay, 260 km south of Lima, Perú, along the entire Chilean continental coast, Argentinean Patagonia to northern Argentina (see Schrödl 2003). This is the first record of this otherwise common species from the Chonos Archipelago. Observations. Living specimens ranged between 40 and 50 mm body length. They did not show the network of more or less dark brown pigment between the tubercles that is characteristic for most northern and central Chilean specimens (see Schrödl 2003); instead, the uniformly yellow coloured specimens externally resemble A. fontaini described from the Comau Fjord (Schrödl et al. in press), the Magellan Strait and Argentina (see Schrödl 2003). Yellow coloration, large tubercles, triangular, grooved oral tentacles, the arrangement of the reproductive system, and the structure of genital organs agree exactly with recent re-descriptions and diagnoses of A. fontaini by Schrödl (1997b, 2000, 2003). A phylogenetic analysis of cryptobranch doridoideans by Valdés (2002) indicates the genus Archidoris and some other traditional dorid genera may form a clade that he called Doris; however, statistic support for such a clade is still lacking, and synonymizing dorid genera thus may be premature. Diaulula Bergh, 1880: Diaulula punctuolata (D’Orbigny, 1837) (Fig. 3A and 3B) Material. Two specimens (ZSM Moll 20040984; both dissected), Ipún Island (44° 33.1’ S, 74° 48.0’ W); found in a small tide pool together with R. pulchra. Distribution. Valparaíso, central Chile, to Melinka, Guaitecas Islands; Magellan Strait, Falklands and Argentinean Patagonia to northern Argentina (see Schrödl 2003). This is the first record of this otherwise common species from the Chonos Archipelago. 5 Observations. Living specimens measured 25 mm and 35 mm in length. External features of the specimens examined, like the uniform whitish coloration, the slender, similar sized caryophyllid tubercles, the six tripinnate to quadripinnate gills, the elevated rhinophoral and branchial sheaths, digitiform oral tentacles, and the bilabiate anterior foot edge with the upper lip notched, confirm exactly with recent redescriptions and diagnoses of D. punctuolata by Valdés & Gosliner (2001), Valdés & Muniaín (2002), and Schrödl (2003). While the smaller specimen was immature, the larger one had its reproductive system fully developed, except for the still relatively smallsized female gland mass. Rostanga Bergh, 1879: Rostanga pulchra MacFarland, 1905 (Fig. 4A, 4B; 5A-5D) Material. One specimen (ZSM Moll 20040981), Ipún Island (44° 33.1’ S, 74° 48.0’ W); found in a small tide pool together with D. punctuolata. Distribution. Alaska to Mexico; Bahía de Coliumo to northern Chiloé; Bahía Camarones, Argentina (see Schrödl 2003). This is the first record of R. pulchra from the Chonos archipelago extending its known geographic range in the Pacific by approximately 300 km to the south. Description. The single living specimen was 6 mm long and uniformly orange coloured (Fig. 4A and 4B). The preserved specimen (6 mm) shows a dorsum densely covered by slender caryophyllid tubercles with a height up to 200 µm and measuring 50-100 µm in diameter. The apical sensory knob is elongated, it measures approximately half the diameter of the corresponding tubercle. The tip of the tubercle is crown-like surrounded by five to six vertical needle-like spicules. Most of the tubercles are, however, artificially eroded apically. Within the notum there is a layer of similar shaped horizontally arranged spicules. There are 10 unipinnate gills surrounding the anal papilla. The rhinophores bear approximately 10 vertical lamellae on each side below a large knobby clavus. The notum does not bear branchial or rhinophoral sheaths. The oral tentacles are short conical. The foot is broad and anteriorly bilabiate, the upper lip is notched. The jaws are small, solid, with some rows of elongated, overlapping rodlets. The radula formula is approx. 80 x 30.30.0.30.30. The rachis is broad. 6 SCHRÖDL & GRAU Fig. 2: Archidoris fontaini. (A) Two living specimens in its natural habitat (dorsolateral view). (B) Living specimen in ventral view. Archidoris fontaini. (A) Dos especímenes vivos en su hábitat natural, vista dorsolateral. (B) Espécimen vivo, vista ventral. Fig. 3: Diaulula punctuolata. (A) Living specimen in its natural habitat, dorsal view. (B) Living specimen in ventral view. Diaulula punctuolata. (A) Espécimen vivo en su habitat natural, vista dorsal. (B) Espécimen vivo, vista ventral. Fig. 4: Rostanga pulchra. (A) Living specimen in its natural habitat, dorsal view. (B) Close-up of living specimen, dorsal view. Rostanga pulchra. (A) Espécimen vivo en su hábitat natural, vista dorsal. (B) Acercamiento de espécimen vivo, vista dorsal. NUDIBRANCHIA FROM SOUTHERN CHILE 7 Fig. 5: Rostanga pulchra, SEM micrographs of radular structures. (A) Innermost lateral teeth. (B) Midlateral teeth. (C) Outer lateral teeth. (D) Marginal teeth. Rostanga pulchra, fotomicrografías de las estructuras radulares. (A) Dientes laterales internos. (B) Dientes centro-laterales. (C) Dientes laterales externos. (D) Dientes marginales. Lateral and marginal teeth are densely arranged and there is considerable overlap between different rows, thus inhibiting accurate counting of maximum teeth numbers. First lateral teeth are small, hook-shaped, with a stout cusp that is serrate with up to eight pointed denticles at its inner edge (Fig. 5A). The number, length and arrangement of denticles is variable between teeth of different rows. Inner lateral teeth are hook-shaped with a broad base bearing a blunt protuberance (denticle) (Fig. 5B). Midlateral teeth grow successively in size with the hook becoming longer and more slender. Outer laterals gradually become slender and elongate (Fig. 5C). Marginal teeth are very thin and elongate, splitting into a fine brush of several denticles in the upper third (Fig. 5D). The esophagus is a straight thin tube entering the digestive gland anteroventrally. Both stomach and caecum are embedded within the digestive gland. The intestine forms a loop and leads posteriorly towards the anal papilla as a thin tube. The preservation condition of the specimen did not allow anatomical examination of further digestive, circulatory and excretory features. The cerebropleural ganglia are completely fused. Eyes are sessile. There is a small ganglion associated to the cerebral ganglion that might be a rhinophoral ganglion. The ampulla is long tube-like and filled with sperm. The prostate is an irregularily spherically shaped granular mass. The vas deferens is thin and muscular. The genital openings, female gland mass and allosperm receptacles are poorly developed, the specimen was not fully mature. SCHRÖDL & GRAU 8 DISCUSSION Remarks on the taxonomy of Rostanga pulchra Rostanga pulchra was originally described from California by MacFarland (1905). Muniaín & Valdés (2000) compared additional specimens of R. pulchra from the type locality, Monterey, California, with the similar, western Atlantic Rostanga byga Marcus, 1958; of the latter, the dissected holotype from Brazil was re-examined and additional specimens from Argentina were studied anatomically in detail. Due to a number of external and anatomical differences Muniaín & Valdés (2000) concluded that R. pulchra is clearly distinct from R. byga. However, most of these differences only refer to the specimens examined by themselves. Original descriptions of R. pulchra by MacFarland (1905, 1906, 1966) and R. byga by Marcus (1958) were not considered properly, South American specimens of R. pulchra were not included into their analyses. On one hand, Muniaín & Valdés (2000) considered previous records of R. pulchra, i.e., from Bahía Camarones (44°29’ S), southern Argentina by Marcus & Marcus (1969), from the central Chilean Bahía de Coliumo, from Queule (north of Valdivia), and from the northern coast of Chiloé Island (Schrödl 1996, 1997, 2003), to be problematic due to the lack of detailed anatomical descriptions. On the other hand, Muniaín & Valdés (2000) accepted specimens from Chiloé Island described in great detail by Marcus (1959) as being R. pulchra due to sharing two “characteristic” radula features: (1) the innermost radular teeth having a short cusp in relation to their denticles, and (2) mid-lateral teeth having a large secondary cusp. However, the shape of innermost laterals was illustrated to be very variable by Marcus (1959: Fig. 66A and 67). There is no indication of any secondary cusps in mid-lateral teeth in Marcus’ specimens (but Marcus 1959 did mention the second and third laterals to have basal swellings in his German text description). Several further radular and reproductive features (see Table 1) of Marcus’ Chilean specimens were not considered by Muniaín & Valdés (2000) but also differ considerably from conditions they regarded diagnostic for (in fact only their own Monterey specimens of ) R. pulchra. The shape of midlateral teeth of Marcus’ specimens, as well as the shape of the ampulla, and the arrangement of allosperm receptacles which, according to Marcus (1959), are “like in R. byga”, all resemble more to conditions stated for R. byga than to Californian R. pulchra examined by Muniaìn & Valdés (2000). Thus, the anatomy and taxonomy of South American R. pulchra was in need to be re-evaluated. Table 1 summarizes taxonomically relevant external and anatomical knowledge of specimens assigned to R. pulchra, including new external and radula data presented herein. According to present, still limited knowledge, there are not any differences amongst Chilean specimens. Muniaín & Valdés (2000) in particular doubted conspecifity of a specimen collected off Bahía Camarones, Argentina, due to its many (approximately 90) radula teeth per half row. This is, however, exactly within the range of central and southern Chilean R. pulchra (see Marcus 1959, Schrödl 2003; Table 1). According to Table 1, Southern American specimens of R. pulchra resemble northeastern Pacific R. pulchra described by MacFarland (1905, 1966) regarding all external, radular and reproductive features examined. The specimens from Monterey Bay described by Muniaín & Valdés (2000) differ in having (1) less radula rows and teeth per half row, (2) due to the presence (vs. absence) of a long denticle in inner and mid-lateral teeth, and (3) having marginal teeth splitted into nine to 10 (versus one to six) brush-like denticles. In addition to intraspecific variability, the exact numbers of radula rows and teeth per half-row are, however, difficult to count due to dense arrangement and considerable overlap. The specimen examined herein does not show any long basal denticles in inner lateral teeth but a broad, blunt elevation (“low denticle”) at the base of midlateral teeth. Besides varying within teeth of the same individual, the exact number of brush-like denticles of marginal teeth (Fig. 5D) is difficult to count even if using SEM. With present knowledge, northern and southern hemispherical populations of R. pulchra cannot be consistently distinguished morphologically; radula differences amongst specimens from California include the range of variability known from Chilean specimens. NUDIBRANCHIA FROM SOUTHERN CHILE Comparision between Rostanga pulchra and R. byga According to Muniaìn & Valdés (2000), R. pulchra and R. byga can be clearly distinguished. However, besides color differences (absence vs. presence of white spots on the central notum), their Table 1 does not show any consistent distinguishing feature. The radula formulae overlap with regard to number of rows and teeth per half row. Chilean and Argentinian R. pulchra were described to possess up to nine and seven denticles on the innermost radular teeth, respectively (Marcus 1959, Marcus & Marcus 1969), while six to 11 were mentioned for Californian R. pulchra, and five to 10 for R. byga specimens by Marcus (1958) and Muniaín & Valdés (2000). Innermost radula teeth of Chilean R. pulchra were already shown to be very variable in shape by Marcus (1959: figures 66A, 67). According to SEM photographs of Muniaín & Valdés (2000), we cannot see any difference between a “short” or “long” cusp of the innermost (= first) lateral teeth of R. pulchra and R. byga. The long denticle that is present in R. pulchra specimens from Monterey examined by Muniaín & Valdés (2000) is absent in Californian specimens described by MacFarland (1905, 1966) and in South American specimens described by Marcus (1959). A broad and low basal elevation is present in inner and mid-lateral teeth of the specimen from Ipún Island examined herein and in R. byga (see Muniaín & Valdés 2000). Two areas with several rows of jaw rodlets are present in all R. pulchra specimens studied and were described from the holotype of R. byga as well (Marcus 1958), while there were only few rodlets in R. byga studied by Muniaín & Valdés (2000). Allosperm receptacles are arranged serially in both R. pulchra and R. byga. According to Marcus (1959) and results presented in this paper, the ampulla of Chilean R. pulchra does not differ in its tubular shape from that of R. byga. The sensory knob of the caryophyllid tubercles is “extremely small” in R. pulchra from Monterey and “large” in R. byga (Muniaín & Valdés 2000), while measuring approximately half the diameter of the tubercles in R. pulchra from Ipún and in a specimen from the Bay of Coliumo (ZSM 19960719) re-examined herein. This character should be reinvestigated using appropriately preserved material. 9 Thus, with present knowledge (Table 1), the diffences between R. pulchra and R. byga are limited to (1) the absence versus presence of white dots on the central notum, (2) nine to 12 versus 12-16 (i.e., not “twice as many” as stated by Muniaín & Valdés) rhinophoral lamellae, (3) the insertion into the bursa copulatrix of the vagina and the duct leading towards the receptaculum seminis that appears to be more separated in R. pulchra, and, (5) the receptaculum seminis that is stalked in R. byga while it is not (or very shortly) stalked in R. pulchra. Different sizes and proportions of reproductive organs such as the apparently larger prostate of R. byga should be interpreted with caution since they also may depend on the maturity of individuals. We follow Marcus (1958) stating that R. pulchra and R. byga are very similar. A few minor but consistent distinguishing features listed above indicate that they are distinct species. Additional evidence comes from geographical and hydrographical indications: while R. pulchra appears to be a bipolar species known from cold-temperate waters of the northeastern and southeastern Pacific and from southern Argentina, R. byga is obviously adapted to warmer and tropical waters; it ranges from northern Brazil south to the Gulf of San José (approximately 42°20’ S, Muniaín & Valdés 2000), i.e., just north of the Península Valdez, the virtual border to the Magellanic region. Biogeography The new records of A. fontaini and D. punctuolata from Guamblin and Ipún islands represent new localities for these widespread Magellanic species; in the Chilean archipelagos remains a gap between 45 and 52° S in their elsewhere continuous geographic record along the Chilean and Argentinean coasts (Schrödl 1997a, 2003). The finding of Rostanga pulchra at Ipún Island represents the southernmost record of this species in the Pacific. Schrödl et al. (in press) showed further, supposedly warm-adapted species to occur in the southern Chilean fjordland. Like R. pulchra, Cadlina sparsa (Odhner, 1921) is a bipolar eastern Pacific and amphi-South American species, with a geographic record extended south to the Comau fjord system (42.1– 42.5° S) recently. The amphi-South American Doto uva Marcus, 1955, the Peruvian Phidiana Jaw rodlets Body length of specimens examined anatomically Two areas with six Two areas with Two areas with slightly rough pegs rows of about 20 five-to-six rodlets each overlapping rows of rodlets each 6 mm (preserved) Text: “jaws solid, Elongate rodlets with few rodlets of varying lengths”. SEM micrograph showing over 50 rodlets 16 and 18 mm (preserved) Two areas with several rows of elongate rodlets 6 mm (preserved) Elongated, approx. half diameter of corresponding tubercle Extremely small and ciliated, with small marginal cilia Shape of sensory knob “A number of” 15 mm (preserved) Five-to-six Four-to-five Five-to-six Number of spicules surrounding sensory knob 9 mm (preserved) Height up to 200 µm; 50-100 µm in diameter Cup-shaped, height Slender about 150 µm Height up to 250 µm; 50-80 µm in diameter Dimension and shape Height up to of caryophyllid 420 µm; up to tubercles 80 µm in diameter Vertical; 10-12 lamellae Vertical; approximately 10 lamellae Arrangement and number of rhinophoral lamellae Vertical, transverse; Vertical; 10-12 nine lamellae lamellae n=1 Vertical; 10-12 lamellae n = 18 Bahía de Coliumo, Ipún Island, Chile Queule, northern coast of Chiloé, Chile This study Orange n=6 Monterey Bay, USA Muniaín & Valdés Schrödl (2003) (2000) Orange-yellow Orange to red, to bright red, some some specimens specimens with with small dark dark spots scattered spots on the dorsum; small white dots on mantle margin Light yellow- Preserved specimen Orange red; one with some dark preserved red to deep spots specimen with scarlet, with dark subepidermal brown or black spots spots on the dorsum Coloration n=3 Many Specimens n=1 California, USA Northern coast of Chiloé Island, Chile Marcus (1959) Bahía de Camarones, Argentina Marcus & Marcus MacFarland (1906), (1966) (1969) R. pulchra Collecting locality Reference Feature or character Comparación de Rostanga pulchra MacFarland, 1905, con R. byga Marcus, 1958 Comparison of Rostanga pulchra MacFarland, 1905, and R. byga Marcus, 1958 TABLE 1 Two small areas with six rows, each with about 15 rodlets 11 mm (preserved) Varying sizes Vertical, 12 lamellae Bright brick red with white spots on some tubercles n=1 Ilhabela, Brazil Marcus (1958) Very small; with few irregular rodlets 12 and 14 mm (preserved); 20 mm (alive) Large, rounded, ciliated; with small marginal cilia Six Height about 100 µm Vertical, transverse; 14-16 lamellae Orange, lightly spotted with white dots in the middle of the dorsum and between tubercles n=3 Gulf of San José, Argentina Muniaín & Valdés (2000) R. byga 10 SCHRÖDL & GRAU Hook-shaped, with broad base Inner lateral teeth Very short stalk or Very short stalk inserted jointly Shortly stalked Insertion of receptaculum seminis Very short stalk or inserted jointly Gradually increasing in size, cusps becoming more slender and longer Vagina separated from duct leading to receptaculum seminis Vagina separated from duct leading to receptaculum seminis Vagina separated from duct leading to receptaculum seminis Arrangement of Vagina allosperm receptacles separated from duct leading to receptaculum seminis Long stalk Bursa inserted nearly jointly by vagina and duct leading to receptaculum seminis Large Long and tubular Slender tube Well developed, but bursa copulatrix not yet fully developed Very thin and elongate, almost apical half forming a fine brush of five-to-nine denticles Long and slender, upper third split into several brushlike denticles Larger than bursa copulatrix Very thin and elongate, with brush-like apical denticles Similar sized as bursa copulatrix Large Thin and elongate, with a fine brush of nine-10 denticles at the end Prostate Long and slender, upper third split into one-to-four additional denticles Small, hook-shaped; base larger than cusp Second to 30 th Gruadally teeth growing in increasing in size, size and becoming cups becoming slender and erect; more slender and with broad and low longer Marginal teeth basal denticle Hook-shaped, with broad base and broad and low denticle Long and slender tube Apical fourth split Long and into up to six slender, spinelike cusps distally split into one-to-six additional, very long denticles Second to 30th teeth Teeth gradually growing in size and growing in size, secondary cusp well becoming more developed but slender and erect becoming smaller in relation to the long primary cusp Broad wing-like base Stalked Bursa inserted nearly jointly by vagina and duct leading to receptaculum seminis Very large, flat, with two welldifferentiated portions Long and tubular Very thin and elongate, with a fine brush of sixto-10 denticles at the end Gruadally increasing in size Broad triangular base with a small lateral wing, pointed primary cusp 45-62 x 5362.0.53-62 Hook-shaped, with Wide, with a long two basal swellings pointed primary cusp and a large pointed secondary cusp Approximately Approximately 80 x 30.30.0.30.30 60 x 60.0.60 Short; rounded, “long” cusp with seven-to-10 short denticles 68 x 85.0.85 Short; “short” cusp Short, with four-to- Small and stout; up Short; broad cusp Small and stout; to eight more or less with five-to-eight denticles variable with six-to-eight eight denticles denticles in shape and elongate denticles denticles number: either four long comb-like apical denticles or serrate cusp with nine small denticles 50-53 x 4851.0.48-51 Ampulla Marginal teeth Mid-lateral and outer Hook grows in lateral teeth size and becomes more slender and less curved Usually three-toShort, cusp serrate with 20- five, up to seven 30 or eight-to- denticles 11 denticles (only six denticles drawn in 1966: plate 35, Fig. 1) Shape of 1st lateral teeth 85 x approximately 80 x 90.0.90 90.0.90 60-68 x 6081.0.60-81 Radula formula NUDIBRANCHIA FROM SOUTHERN CHILE 11 SCHRÖDL & GRAU 12 lottini (D’Orbigny, 1837), and the central Chilean Tergipedidae sp. (Schrödl 2003) were collected from the Comau fjord as well (Schrödl et al. in press). In addition, Doto uva was found at Puerto Cisnes (44.7° S) (Schrödl et al. in press), and Phidiana lottini was photographically recorded from Guaitecas Islands close to Melinka (Schories, unpublished information). With R. pulchra and D. uva, at least two of five nudipleuran species classified as “warm (temperate) amphi-South American” (Schrödl 2003) now are known from considerably south of the Chiloé Island area, that should not be regarded as a strict distributional barrier for nudipleurans any longer. Regarding other marine invertebrates, e.g. sea anemones, the Península Taitao at 46° S appears to be an area of faunal change between cold- and warm-temperate species (Lancellotti & Vásquez 1999, Häussermann & Försterra in press). Further collects in the southern Chilean archipelago and fjordland will be necessary to unravel nudibranch diversity and their actual distributional limits. ACKNOWLEDGEMENTS Field work was part of an expedition carried out with help of Corporación Nacional Forestal; many thanks to Jorge Valenzuela Rojas for letting JG participate. This study derived from a university course on “Systematics and Biology of Chilean Opisthobranchia” held at the Universidad Austral de Chile (UACh), Valdivia, that was financed by a guest lecturer stipend (MECESUP AUS 0101 grant, organized by Dr. C. Gallardo, directed by Dr. C. Bertrán) to MS; many thanks to the Zoology Institute for working facilities and friendly support. This study was supported by the GeoBioCenterLMU. LITERATURE CITED BRATTSTRÖM H & A JOHANSSEN (1983) Ecological and regional zoogeography of the marine benthic fauna of Chile. Report No 49 of the Lund University Chile Expedition 1948-49. Sarsia 68: 289-339. HÄUSSERMANN V & FÖRSTERRA (in press) Distribution patterns of Chilean shallow-water sea anemones (Cnidaria: Anthozoa: Actiniaria, Corallimorpharia); with a discussion of the Associate Editor: Patricio Camus Received October 21, 2004; accepted July 15, 2005 taxonomic and zoogeographic relationships between the actinofauna of the South East Pacific, the South West Atlantic and Antarctica. Scientia Marina, supplement. LANCELLOTTI D A & J A VÁSQUEZ (1999) Biogeographical patterns of benthic macroinvertebrates in the Southeastern Pacific littoral. Journal of Biogeography 26 1001-1006. MacFARLAND F M (1905) A preliminary account of the Dorididae of Monterey Bay, California, and vicinity. Proccedings of the Biological Society of Washington (USA) 18: 35-54. MacFARLAND F M (1906) Opisthobranchiate Mollusca from Monterey Bay, California, and vicinity. Bulletin of the United States Bureau of Fisheries (USA) 25: 109-151, plates 18-31. MacFARLAND F M (1966) Studies of opisthobranchiate mollusks of the Pacific coast of North America. Memoirs California Academy Sciences 6: 1-546, plates 1-72. MARCUS Er (1958) On western Atlantic opisthobranchiate gastropods. American Museum Novitates 1906: 1-82. MARCUS Er (1959) Lamellariacea und Opisthobranchia. Reports of the Lund University Chile Expedition 1948-49, No 36. Lunds Universitets Arsskrift, N. F., 55: 1-133, figures 1-196. MUNIAÍN C & A VALDÉS (2000) Rostanga byga Er. Marcus, 1958 from Argentina: redescription and comparision to Rostanga pulchra MacFarland, 1905 (Mollusca, Nudibranchia, Doridina). Proceedings of the California Academy of Sciences (USA) 52: 1-10. SCHRÖDL M (1997a) Range extensions of Magellanic nudibranchs (Opisthobranchia) into the Peruvian faunal province. Veliger 40: 38-42. SCHRÖDL M (1997b) On the morphology of the Magellanic nudibranch Anisodoris fontaini (D’Orbigny, 1837), and its synonymy with A. tessellata Bergh, 1898. Veliger 40: 228-233. SCHRÖDL M (2000) Taxonomic revision of the common South American nudibranch Anisodoris fontaini (D’Orbigny, 1837), with discussion of its systematic placement. Journal of Molluscan Studies 66: 49-61. SCHRÖDL M (2003) Sea slugs of southern South America. ConchBooks, Hackenheim, Germany. 165 pp. SCHRÖDL M, M-A ALARCÓN, LR BEDRINANA, FJ BRAVO, CM BUSTAMANTE, R CARVALHO, G FÖRSTERRA, C GALLARDO, V HÄUSSERMANN & A SALMEN (in press) Nudipleura (Gastropoda: Opisthobranchia) from the southern Chilean Comau Fjord, with redescription of Polycera priva Marcus, 1959. Vita Malacologica (Holland). VALDÉS Á (2002) A phylogenetic analysis and systematic revision of the cryptobranch dorids (Mollusca, Nudibranchia, Anthobranchia). Zoological Journal of the Linnean Society 136: 535-636. VALDÉS Á & TM GOSLINER (2001) Systematics and phylogeny of the caryophyllidia-bearing dorids (Mollusca, Nudibranchia), with descriptions of a new genus and four new species from Indo-Pacific deep waters. Zoological Journal of the Linnean Society 133: 103-198. VALDÉS Á & C MUNIAÍN (2002) Revision and taxonomic reassessment of magellanic species assigned to Anisodoris Bergh, 1898 (Nudibranchia: Doridoidea). Journal of Molluscan Studies 68: 345-351.
https://openalex.org/W2475092848	https://www.scielo.br/j/bjce/a/Rs8VnqTcfgVp3Y8nZZY8dkg/?lang=en&format=pdf	English	null	ADVANCED CONTROL OF A COMPLEX CHEMICAL PROCESS	Brazilian Journal of Chemical Engineering/Brazilian journal of chemical engineering	2,016	cc-by	11,135	(Submitted: July 25, 2011 ; Revised: February 12, 2015 ; Accepted: February 21, 2015) Abstract - Three phase catalytic hydrogenation reactors are important reactors with complex behavior due to the interaction among gas, solid and liquid phases with the kinetic, mass and heat transfer mechanisms. A nonlinear distributed parameter model was developed based on mass and energy conservation principles. It consists of balance equations for the gas and liquid phases, so that a system of partial differential equations is generated. Because detailed nonlinear mathematical models are not suitable for use in controller design, a simple linear mathematical model of the process, which describes its most important properties, was determined. Both developed mathematical models were validated using plant data. The control strategies proposed in this paper are a multivariable Smith Predictor PID controller and multivariable Smith Predictor structure in which the primary controllers are derived based on Internal Model Control. Set-point tracking and disturbance rejection tests are presented for both methods based on scenarios implemented in Matlab/SIMULINK. p p Keywords: Multivariable Smith Predictor; Internal Model Control; Hydrogenation process; Dynamic mathematical model. Brazilian Journal of Chemical Engineering ISSN 0104-6632 Printed in Brazil www.abeq.org.br/bjche Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 dx.doi.org/10.1590/0104-6632.20160331s00001267 To whom correspondence should be addressed Roxana Both1, Eva-Henrietta Dulf1 and Ana-Maria Cormos2 1Technical University of Cluj Napoca, Memorandumului St., No. 28, RO - 400014, Cluj-Napoca, Romania. 2Babes-Bolyai University, Faculty of Chemistry and Chemical Engineering, Arany Janos St., No. 11, RO-400028, Cluj-Napoca, Romania. Phone: + 40-264-401821; Fax: + 40-0264-401220 E-mail: Eva.Dulf@aut.utcluj.ro; http://aut.utcluj.ro Brazilian Journal of Chemical Engineering Brazilian Journal of Chemical Engineering INTRODUCTION variable Smith Predictors is the decoupling of the process. The industrial scale 2-ethyl-hexenal hydrogenation process produces 2-ethyl-hexanol oxo-alcohol, used as raw material for dioctyl-phthalate (DOP) or 2- ethylhexyl-phthalate (DEHP) production. DOP and DEHP represent around 50% of the global consump- tion in the production of poly(vinyl chloride) (PVC) (ICIS,2011;WRI,2009;SRI,2011). The present paper is structured in 8 parts. After the introduction, the second part describes the hydro- genation process and also details the actual control and the new proposed control system. The third part presents the developed dynamic mathematical models of the hydrogenation process: the nonlinear distrib- uted parameter mathematical model and the linear simplified mathematical model. Both models are validated using plant data, acquired courtesy of S.C. Oltchim S.A., Ramnicu Valcea, Romania. In the control algorithm design procedure for multi-input multi-output (MIMO) systems, it is necessary a measure of interactions within MIMO systems, which can be provided by the Relative Gain Array (RGA) analysis presented in section four. Sections Because the hydrogenation reaction is highly exo- thermic, the reactor output temperature is of great importance. Like most of chemical and petro-chemi- cal plants, the 2-ethyl-hexenal hydrogenation process presents large time delays and is multivariable by nature. For multivariable time delay processes, the control solution will revolve around a multivariable Smith Predictor. A very important step for the multi- Rooxana Both, Eva- -Henrietta Dulf and Ana-Maria Coormos 156 wh hex hex here: A is 2- xanal (inter xanol (final p -ethyl-hexena rmediate pro product). al (reactant), oduct) and , B is 2-ethy C is 2-ethy yl- yl- fi d se tw re ive and six esign, respe even the com wo control st epresent the present a m ctively IMC mparative sim trategies are last part of th multivariable C control des mulation resu presented. T he paper. PID contro sign. In sect ults between The conclusio ller tion the ons sec pro tem rat hex to exo kca sid rea ord cal the lys tai dil its flo pro The main p cutive hydro oduct being mperature (i te and purity xenal). The t the fact that othermic wi al/kmol (Sm de-reactions, actants betw der to start th l recommend e range of 16 st degree of n the tempe lution of the elf (the refl ow diagram ocess is pres parameters th ogenation re g 2-ethyl-hex input/output) y of input rea temperature t the hydrog ith a reacti melder,1989). PROCESS DESCRIPTION p p m si se p p ch d v co th re su v th sc ch th The 2-ethy lex, which ca hase in the main product ide-products econdary si resent both hase 2-ethy haracterized imensions o antage: the p omparison to he advantag eaction of 2 umption, sm olumes of ca he impossibi cale liquid p hemical indu he present wo yl-hexenal hy an take place presence of t (2 ethyl-hex like: n-but ide-reactions advantages a yl-hexenal h by high ene of equipment possibility to o the gas pha es of the li 2-ethyl-hexe mall dimensio atalyst. How ility of catal phase hydrog ustries and re ork. ydrogenation e both in liqu a catalyst a xanol), can a tanol or iso s. Technolog and disadvan hydrogenatio ergy consum t, but has an o regenerate ase hydrogen iquid phase nal are: low on of equipm wever, the ma lyst regenera genation is p epresents the process is co uid phase or and, besides also form ot o-butanol fr gical solutio ntages. The on reaction mption and gr n important the catalyst. nation reacti hydrogenat w energy c ment, and sm ain drawback ation. Indust predominant e main focus om- gas the ther rom ons gas is reat ad- . In ion, tion on- mall k is rial t in s of Fig hex pla gure 1: Proc xenal hydro ants. cess flow dia ogenation pr agram of a ty rocess used ypical 2-ethy in industri yl- ial p n se fo re An import layed by the ickel catalys en based on or the 2-ethy ecommended tant role in th e catalyst. Fo st deposited the BASF li yl-hexenal hy d in specific he hydrogena or the presen on silica su icense. This ydrogenation literature (Sm ation reaction nt case study upport was c type of catal reaction is a melder, 1989 n is y, a ho- lyst also 9). (2 n p 2 to fr tr fr re th m o ly is Several res 2011) studie ism. INTRODUCTION but also an ween 90°C an he reaction. T ded temperat 60 °C – 180 ° activity. The erature below 2-ethyl-hexa ux rate). A of the 2-eth ented in Figu hat influence eactions (the xanal) are: ), reactor lo actants (hydr is a critical genation reac on heat of . High temp n input temp nd 110°C is The maximu ture inside th °C, dependin e solution ch w the critica anal flow wi typical indu hyl-hexenal h ure 1. e the two co e intermedia the pressur oading, reflu rogen, 2-ethy parameter du ction is high 25.27 x 1 peratures fav perature of th s necessary um technolog he reactor is ng on the cat hosen to mai al value is th ith the produ ustrial proce hydrogenatio on- ate re, ux yl- ue hly 03 vor he in gi- in ta- in- he uct ess on R C H CR C OH + H 2 R C H 2 C HR H 2 + H C OH R C H 2 C HR C H 2 O H Distributed Parameter Nonlinear Mathematical Model At the present time, the hydrogenation reactor is operated using an open loop control system, so the main parameters like 2-ethyl-hexenal input flow rate, recirculated 2-ethyl-hexanol flow and input tem- perature of the reactants are maintained at the im- posed values, without feedback control of output temperature or 2-ethyl-hexanol output concentra- tion. This control solution is characterized by a main disadvantage: it is not fault-tolerant, being difficult to apply correction methods if one control loop fails. However, the hydrogenation reactor will operate in a satisfactory manner if the auxiliary control loops for flow control of the reactants and input temperature control of the reactants are work- ing in good parameters. An accurate mathematical model is needed to evaluate the operational challenges and to understand the processes that occur inside the reactor and also to develop a more efficient control strategy. In the spe- cific literature, a number of kinetic studies of the hydrogenation reaction are reported (Smelder, 1989; Niklasson, 1987, 1988). However, no mathematical model of the process is given. Motivated by these reasons, in previous works (Both, 2013), a first principle based mathematical model was developed starting only from some equa- tions describing the hydrogenation reaction kinetics. The developed model (Both, 2013), consisting of numerous partial differential equations and several algebraic ones, was implemented and validated using plant data acquired courtesy of S.C. Oltchim S.A. This model is useful in the design, optimization and operation of the chemical reactor and especially in control design and testing. The reaction rate equa- tions, transport models, energy and mass balances were coupled and were solved with respect to time and space with algorithms suitable for partial differen- tial equations (Attou et al., 1999; Burghardt et al., 1995, Iliuta et al., 1997). At present an operator sets manually the reference values for the 2-ethyl-hexenal input flow rate, recir- culated 2-ethyl-hexanol flow and input temperature of the reactants. The 2-ethyl-hexanol output concen- tration is measured periodically and, if it is not satis- factory, the reference values for the main input varia- bles are adjusted by the accumulated expertise of the operator. In order to optimize the production, the authors propose a feedback control system using the existing infrastructure. The implementation costs are reduced to the costs of a temperature transducer used to measure the output temperature of the product. MATHEMATICAL MODELS the flow ratio (the recirculated 2-ethyl-hexanol flow must be decreased) and also to increase the input temperature of the reactants to start the hydrogena- tion reaction. PROCESS DESCRIPTION Hence, resent case 011): the tra o the gas-liqu rom the gas- ransport of r rom the liqu eactant adsor he chemical molecules; pr f the catalys yst surface to s expressed a p searchers lik ed the hydro the reaction study are (B ansport of hy uid interface -liquid interf reactants 2-e uid phase to rption to the l reaction b roduct desor st; the produ o the liquid p as follows (C ( ke Bozga (20 ogenation re n mechanism Bozga et al., ydrogen from e; the transpo face to the li ethyl-hexenal o the liquid- active sites between ads rption from uct transport phase. The re Collins et al., 001) and Coc eaction mec m stages for , 2001; Cock m the gas ph ort of hydrog quid phase; l and hydrog solid interfa of the cataly sorbed react the active si from the ca eaction pathw 2009): ) cker cha- the ker, hase gen the gen ace; yst; tant ites ata- way Fig hex pla gure 1: Proc xenal hydro ants. cess flow dia ogenation pr agram of a ty rocess used ypical 2-ethy in industri yl- ial Fig hex pla gure 1: Proc xenal hydro ants. cess flow dia ogenation pr agram of a ty rocess used ypical 2-ethy in industri yl- ial po hy tur flo rec tai and me a d rea pro of As presente rtance is the ydrogenation re. It was det ow ratio betw circulated 2 ned to carry d also to ma ended range. disturbance b action and im oduct. The n a decrease ed before, a e reactor loa reaction its termined exp ween the 2-e -ethyl-hexan out the reac aintain the t The catalys because it inf mplicitly the necessary ste in the cataly a parameter ading. This self and also perimentally ethyl-hexenal nol flow mu ction with go temperature st degree of a fluences the h output temp eps to counte yst activity a of great im influences th o the temper that a specif l flow and th ust be mai ood paramete in the recom activity acts hydrogenatio perature of th eract the effe are: to chang m- he ra- fic he in- ers m- as on he ect ge B A B C Brazilian Jouurnal of Chemical Engineering Advanced Control of a Complex Chemical Process 157 Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 Distributed Parameter Nonlinear Mathematical Model The equations describing the nonlinear mathe- matical model used are (Both, 2013):  The total mass balances for the gas and liquid phases are (Both, 2013): In order to ensure safe and normal conditions of operation for the hydrogenation process there are two main goals of the control strategy. The first goal is to maintain the temperature at an acceptable value. This goal can be achieved by assuring a specific volumetric flow ratio between the 2-ethyl-hexenal and 2-ethyl-hexanol flow rates. The recirculated 2-ethyl-hexanol flow rate is considered to be the ma- nipulated variable. The second goal is to ensure a high conversion of 2-ethyl-hexenal to 2-ethyl-hexa- nol. This goal can be reached by assuring the neces- sary input temperature of the reactants to start the reaction. The input temperature of the reactants must be increased as the catalyst degree of activity decreases in time. The input temperature of the reactants also has an important influence on the output temperature. Overall, the two considered control inputs are the 2 ethyl-hexanol flow rate and the input temperature of the reactants. The output temperature and product concentration are consid- ered to be the measured outputs and the input hy- drogen pressure (P), and 2-ethyl-hexenal flow (Qenal) act as disturbances. 2 G L L L L H v H F F v v S M a N t z           (1) 2           G G G G G H v H F F v v S M a N t z (2) (1) (2)  The component mass balances for liquid and gas phases are (Both, 2013):  The component mass balances for liquid and gas phases are (Both, 2013):         G L L H H L v H C C v a N vph t z (3)        G G G H H L v H C C v a N t z (4) 1        enal enal L sol C C v r t z  (5)         G L L H H L v H C C v a N vph t z (3) (3) (4) (5) Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. Distributed Parameter Nonlinear Mathematical Model 155 - 168, January - March, 2016 Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 158 Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 158 1 2 ( )         anal anal L sol C C v r r t z  (6) 2        oct oct L sol C C v r t z  (7)   2 2 2 2 3 1     anal H anal H enal enal anal anal H H k K K C C r K C K C K C (11) (11) 2        oct oct L sol C C v r t z  (7) (7) where Ki is the adsorption equilibrium constant for component i (i: enal, anal, H), Ci concentration of component i and ki rate constant of surface reaction i,(i:1,2).  The heat balances for the liquid and gas phases are (Both, 2013):  The heat balances for the liquid and gas phases are (Both, 2013): The rate of reaction incorporating the catalyst de- activation can be obtained as follows:          R i i L L L pL H r T T v t z c (8)          G G R i i G pG T T H r v t z c (9) (8) , ( )  i d i r ra t (12) (12) (9) (9) where: a is a fraction of active catalyst and ri is the reaction rate of species i. For validation of the developed model a con- siderable number of trials were chosen to compare simulation results with plant data (NIST, 2011; Gas- par et al., 2010; Perry et al., 1999; Tobiensen et al., 2008, Silva et al., 2003) and the two most repre- sentative cases are presented in Table 1, which em- phasizes the small differences between the plant data and simulated values. Table 1: Data validation. Run H2 concentration (kmol/m3) 2-ethyl-hexenal concentration (kmol/m3) 2-ethyl-hexanol concentration (kmol/m3) Outlet liquid temperature (K) sim plant sim plant sim plant sim plant 1 0.078 0.0549 0.0021 0.0020 5.346 5.32 441.6 441.3 2 0.088 0.055 0.0022 0.0023 5.343 5.31 442.1 441.5  The reaction rates are described by (Smelder, 1989): The accuracy of the distributed parameter nonlinear mathematical model of the process is highlighted by the evolutions of 2-ethyl-hexenal, 2-ethyl-hexanol con- centrations and the temperature evolution along the reactor height, presented in Figure 2 and Figure 3.   2 2 1 1 3 1     enal H enal H enal enal anal anal H H k K K C C r K C K C K C (10) y p mathematical model of the process is highlighted by the evolutions of 2-ethyl-hexenal, 2-ethyl-hexanol con- centrations and the temperature evolution along the reactor height, presented in Figure 2 and Figure 3. Figure 2: 2-ethyl-hexenal and 2-ethyl-hexanol concentration profile along the reactor height - RUN 1. Figure 3: Outlet liquid temperature profile along the reactor height - RUN 1. 0 2 4 6 8 10 12 14 16 18 20 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 Reactor height [m] 2 Ethyl-hexenal concentration [kmol/m3] 0 2 4 6 8 10 12 14 16 18 204.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 2 Ethyl-hexanol concentration [kmol/m3] ___ 2 Ethyl-hexenal concentration [kmol/m3] ------ 2 Ethyl-hexanol concentration [kmol/m3] + plant data 0 2 4 6 8 10 12 14 16 18 20 370 380 390 400 410 420 430 440 450 Reactor height [m] Outlet liquid temperature [K] __Outlet liquid temperature [K] __ Plant data   2 2 1 1 3 1     enal H enal H enal enal anal anal H H k K K C C r K C K C K C (10) Figure 2: 2-ethyl-hexenal and 2-ethyl-hexanol concentration profile along the reactor height - RUN 1. Distributed Parameter Nonlinear Mathematical Model where FL and FG are the liquid and gas phase flow, respectively, S is the cross-sectional area, av is the specific gas-liquid contact area, NHG is the flux of hydrogen transferred from gaseous phase to liquid phase, Cenal is the concentration of 2-ethyl-hexenal, Canal is the concentration of 2-ethyl-hexanal, Coct is the concentration of 2-ethyl-hexanol, L H C is the concentration of hydrogen in the liquid phase and G H C is the concentration of hydrogen in the gaseous phase. vph is the transferred hydrogen flow through the liquid film, adjacent to the catalyst pellet, onto its surface (Bozga, 2001) and ri is the rate of surface reaction i and ΔRHi is the reaction heat of reaction i. Operational Mathematical Model 12 2 1 2 ( ) 1    p dist p p K Hf s T s T s (20) 21 2 1 2 ( ) 1    qec dist qec qec K Hf s T s T s (21) 22 2 1 2 ( ) 1    pc dist p c p c K Hf s T s T s (22) 12 2 1 2 ( ) 1    p dist p p K Hf s T s T s (20) (20) Due to the fact that nonlinear mathematical models are more accurate, but also too complex for efficient use in controller design, the authors propose another approach, to use a simple operational model of the process which describes its most important proper- ties (Iancu et al., 2010; Simson et al., 2001). 21 2 1 2 ( ) 1    qec dist qec qec K Hf s T s T s (21) 22 2 1 2 ( ) 1    pc dist p c p c K Hf s T s T s (22) 21 2 1 2 ( ) 1    qec dist qec qec K Hf s T s T s (21) (21) 22 2 1 2 ( ) 1    pc dist p c p c K Hf s T s T s (22) (22) The linear operational mathematical model was determined using experimental identification methods based on plant data, as a transfer function matrix of second order elements between the main input and output variables (Both, 2012): the 2-ethyl-hexanol recirculated flow (Qoct), the input temperature of the reactants (Tin), the output temperature of the product (Tout) and the output concentration of the product (Cout). Following the previous equations, the transfer matrix of the simplified hydrogenation reactor is: Following the previous equations, the transfer matrix of the simplified hydrogenation reactor is: ( ) ( )     p dist Tout Qoct Qenal H s H s Coct Tin P (23) (23) Using the operational mathematical model, the plant data and the simulation results of the nonlinear validated mathematical model, analysis of the dynamical behavior of the simplified model is done. Table 1: Data validation. 0 2 4 6 8 10 12 14 16 18 20 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 Reactor height [m] 2 Ethyl-hexenal concentration [kmol/m3] 0 2 4 6 8 10 12 14 16 18 204.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 2 Ethyl-hexanol concentration [kmol/m3] ___ 2 Ethyl-hexenal concentration [kmol/m3] ------ 2 Ethyl-hexanol concentration [kmol/m3] + plant data (10) Figure 3: Outlet liquid temperature profile along the reactor height - RUN 1. 0 2 4 6 8 10 12 14 16 18 20 370 380 390 400 410 420 430 440 450 Reactor height [m] Outlet liquid temperature [K] __Outlet liquid temperature [K] __ Plant data Figure 3: Outlet liquid temperature profile along the reactor height - RUN 1. 0 2 4 6 8 10 12 14 16 18 20 370 380 390 400 410 420 430 440 450 Reactor height [m] Outlet liquid temperature [K] __Outlet liquid temperature [K] __ Plant data 0 2 4 6 8 10 12 14 16 18 20 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 Reactor height [m] 2 Ethyl-hexenal concentration [kmol/m3] 0 2 4 6 8 10 12 14 16 18 204.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 2 Ethyl-hexanol concentration [kmol/m3] ___ 2 Ethyl-hexenal concentration [kmol/m3] ------ 2 Ethyl-hexanol concentration [kmol/m3] + plant data Figure 3: Outlet liquid temperature profile along the reactor height - RUN 1. Figure 2: 2-ethyl-hexenal and 2-ethyl-hexanol concentration profile along the reactor height - RUN 1. Brazilian Journal of Chemical Engineering Advanced Control of a Complex Chemical Process 159 Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 Operational Mathematical Model Figure 4 and Figure 5 present the evolution of the output temperature (Tout) for a +14% step variation of the 2-ethyl-hexenal input flow and for a step variation in the 2-ethyl-hexanol recirculated flow. 11 12 21 22 ( ) ( ) ( ) ( ) ( ) qo ti qoc tic s s p s s Hf s e Hf s e H s Hf s e Hf s e              (13) where 11 12 21 22 ( ) ( ) ( ) ( ) ( ) qo ti qoc tic s s p s s Hf s e Hf s e H s Hf s e Hf s e              (13) (13) where 11 2 1 2 ( ) 1    qo qo qo K Hf s T s T s (14) 12 2 1 2 ( ) 1    ti ti ti K Hf s T s T s (15) 21 2 1 2 ( ) 1    qoc qoc qoc K Hf s T s T s (16) 22 2 1 2 ( ) 1    tic tic tic K Hf s T s T s (17) (14) Figure 4: Outlet liquid temperature evolution: change in Qenal. 2600 2800 3000 3200 3400 3600 3800 4000 440 442 444 446 448 450 452 454 Time [s] Tout [K] nonlinear model simplified model 2600 2800 3000 3200 3400 3600 3800 4000 440 442 444 446 448 450 452 454 Time [s] Tout [K] nonlinear model simplified model (15) (16) (17) Figure 4: Outlet liquid temperature evolution: change in Qenal. The input hydrogen pressure (P) and 2-ethyl-hex- enal flow (Qenal) act as disturbances. The transfer matrix describing the dependence between the output variables and the disturbances is: 2600 2800 3000 3200 3400 3600 3800 4000 424 426 428 430 432 434 436 438 440 442 444 Time [s] Tout [K] nonlinear model simplified model ansfer output (18) (19) Figure 5: Outlet liquid temperature evolution: change in Qoct. RELATIVE GAIN ARRAY (RGA) ANALYSIS Considerable research effort was dedicated to the development of control techniques for MIMO pro- cesses, but it still represents a difficult task in practi- cal engineering. At the present time, there are two main directions of approach in control design for multivariable processes. The first approach supports the idea to design a centralized MIMO controller, which implies the use of algebraic decoupling methods or optimal control theory. However, the result is a complex controller, difficult to implement. Another approach is to design a decentralized controller, which attempts to control a fairly multivariable pro- cess by separating the control problem into several SISO (single – input, single - output) systems and then uses conventional control for each loop. The advantages of this approach are very clear: simple to design, easy to implement or tune (fewer parameters); hence it is more often used in practical applications. However, the main disadvantage is represented by the substandard closed loop performance due to in- teractions between loops. A mandatory step is to determine the appropriate loop configuration in order to obtain minimal interactions between loops. This is achieved by proper pairing of the manipulated and controlled variables. In the chemical industry the process complexity reaches up to several hundred control loops, making input – output pairing a diffi- cult task. This task is also essential because it can lead to an unstable overall system, even if the indi- vidual loops are stable. Considerable research effort was dedicated to the development of control techniques for MIMO pro- cesses, but it still represents a difficult task in practi- cal engineering. At the present time, there are two main directions of approach in control design for multivariable processes. The first approach supports the idea to design a centralized MIMO controller, which implies the use of algebraic decoupling methods or optimal control theory. However, the result is a complex controller, difficult to implement. Another approach is to design a decentralized controller, which attempts to control a fairly multivariable pro- cess by separating the control problem into several SISO (single – input, single - output) systems and then uses conventional control for each loop. The advantages of this approach are very clear: simple to design, easy to implement or tune (fewer parameters); hence it is more often used in practical applications. RELATIVE GAIN ARRAY (RGA) ANALYSIS yp In this study the RGA technique was used to de- termine the best control loop pairs and, at the end, the performances of the selected control loop pairs are studied. RGA is represented by a matrix with one column for each input variable and one row for each output variable. From this matrix one can easily com- pare the relative gains associated with each input- output variable pair and match the input and output variables that have the biggest effect on each other, while also minimizing undesired side effects. Standard analysis suggests that RGA elements corresponding to input-output pairings close to 1 should be pre- ferred. Negative or large RGA elements are disad- vantageous as they correspond to loops which may be nominally stable, but which become unstable if saturation occurs (e.g. (Bristol, 1966)). The simplest approach was found to be the use of the developed linear mathematical model and and to compute RGA from the steady-state gain matrix. The steady-state gain matrix is thus obtained with its elements: However, the main disadvantage is represented by the substandard closed loop performance due to in- teractions between loops. A mandatory step is to determine the appropriate loop configuration in order to obtain minimal interactions between loops. This is achieved by proper pairing of the manipulated and controlled variables. In the chemical industry the process complexity reaches up to several hundred control loops, making input – output pairing a diffi- cult task. This task is also essential because it can lead to an unstable overall system, even if the indi- vidual loops are stable.   qo ti qoc tic K K Tout Qoct Coct K K Tin (24) (24) The RGA, for a non-singular square matrix G, is a square complex matrix defined as 1 ( ) ( ) , T RGA G G G   where x denotes element-by element multiplication (Hadamard or Schur product). p In 1996, Bristol (1966) introduced the Relative Gain Array technique to determine a measure of the process interactions, giving advice in solving the pairing problem. This technique considers only the steady-state gains of the process to determine the best pairing solution. Due to its simplicity, the RGA technique was widely used in different applications and still has the widest application in industry. Operational Mathematical Model 2600 2800 3000 3200 3400 3600 3800 4000 424 426 428 430 432 434 436 438 440 442 444 Time [s] Tout [K] nonlinear model simplified model 11 12 21 22 ( ) ( ) ( ) ( ) ( ) qe p qec pc s s dist dist dist s s dist dist Hf s e Hf s e H s Hf s e Hf s e              (18) where 11 2 1 2 ( ) 1    qe dist qe qe K Hf s T s T s (19) 11 12 21 22 ( ) ( ) ( ) ( ) ( ) qe p qec pc s s dist dist dist s s dist dist Hf s e Hf s e H s Hf s e Hf s e              (18) where K 11 12 21 22 ( ) ( ) ( ) ( ) ( ) qe p qec pc s s dist dist dist s s dist dist Hf s e Hf s e H s Hf s e Hf s e              (18) where where 11 2 1 2 ( ) 1    qe dist qe qe K Hf s T s T s (19) Figure 5: Outlet liquid temperature evolution: change in Qoct. 2600 2800 3000 3200 3400 3600 3800 4000 424 Time [s] 11 2 1 2 ( ) 1    qe dist qe qe Hf s T s T s (19) Figure 5: Outlet liquid temperature evolution: change in Qoct. Figure 5: Outlet liquid temperature evolution: change in Qoct. Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 160 The validation of the simplified linear mathemati- cal model is based on the comparison between equa- tion results and plant data. The presence of an ac- ceptable error is emphasized in the previous figures. dynamics, giving a more accurate interaction assess- ment. Operational Mathematical Model Xiong (2005) combined in his work the ad- vantages of both RGA and DRGA by introducing the concept of an effective relative gain array (ERGA), which provides an accurate, detailed and easy to understand description of the dynamic interaction among individual loops without the need to specify the controller type. MULTIVARIABLE SMITH PREDICTOR PID CONTROLLER DESIGN 11 12 21 22 ( ) ( ) ( ) ( ) ( )  R HR s HR s H s HR s HR s (31) 2 2 1 2 1 2 2 2 1 2 1 2 1 1 1 1                   qo ti qoc tic s s qo ti qo qo ti ti p s s qoc tic qoc qoc tic tic K e K e T s T s T s T s H K e K e T s T s T s T s     (28) 2 2 1 2 1 2 2 2 1 2 1 2 1 1 1 1                  qe p qec pc s s qe p qe qe p p s s qec pc qec qec p c p c K e K e T s T s T s T s D K e K e T s T s T s T s     (29) (31) (28) where HR11, HR22 are designed for the direct control of the outputs and HR12, HR21 are designed to com- pensate the coupling effect. Considering the large time delays of the plant the Smith Predictor structure is used, presented in Figure 7. HR11 Hf11 sT e ref1 - + + y1 d1 ref2 HR21 HR12 HR22 Hf21 Hf12 Hf22 sT e sT e sT e Hf11 sT e Hf21 Hf12 Hf22 sT e sT e sT e d2 y2 - - - HR11 Hf11 sT e ref1 - + + y1 d1 ref2 HR21 HR12 HR22 Hf21 Hf12 Hf22 sT e sT e sT e Hf11 sT e Hf21 Hf12 Hf22 sT e sT e sT e d2 y2 - - - Figure 7: Closed loop control scheme using the Smith predictor structure. (29) The hydrogenation process is characterized by the presence of time delays. A typical approach to deal with time delay is the non-delayed output prediction (Melo, 2008; Wang, 2008). The non-delayed output may be estimated and the controller can be computed as for a process without delay. The most popular output predictor is the Smith Predictor, Figure 6. RELATIVE GAIN ARRAY (RGA) ANALYSIS The main disadvantage of this technique is represented by the lack of dynamic information of the process, which may sometimes lead to an incorrect loop pair- ing decision. A dynamic version of the RGA tech- nique was developed by McAvoy (1983). The dy- namic RGA (DRGA), in comparison with the RGA, uses the transfer function model instead of the steady- state gain matrix in order to consider the process For the obtained steady-state gain matrix Hp0 0 2.1 0.0049 1 0.011          Tout Coct p Qoct H Tin (25) (25) and 0 1.2692 0.2692 ( ) 0.2692 1.2692          Tout Coct p Qoct RGA H Tin (26) (26) From the RGA matrix one can observe that there is a strong coupling between Qoct and Tout and Tin and Coct. Brazilian Journal of Chemical Engineering Advanced Control of a Complex Chemical Process 161 MULTIVARIABLE SMITH PREDICTOR PID CONTROLLER DESIGN For the controller design the transfer matrix of the process without disturbance is used: 2 2 1 2 1 2 2 2 1 2 1 2 11 12 21 22 1 1 ( ) 1 1 ( ) ( ) ( ) ( ) qo ti qoc tic qo ti qoc tic s s qo ti qo qo ti ti p s s qoc tic qoc qoc tic tic s s s s K e K e T s T s T s T s H s K e K e T s T s T s T s Hf s e Hf s e Hf s e Hf s e                                    (30) The previously presented linear simplified opera- tional mathematical model of the hydrogenation process consisting of a transfer matrix Hp, which describes the influence of the control inputs u on the controlled outputs y (Tout, Cout), and a transfer ma- trix Hdist, describing the effect of disturbances in the feed z (the input 2-ethyl-hexenal flow Qenal, hydro- gen pressure P) on the outputs y is used in order to develop a multivariable PID controller. (30)    p y H u D z (27) (27)    p y H u D z The desired multivariable controller matrix has the following form [34]: Both matrices Hp and D are (2,2)-matrices. Both matrices Hp and D are (2,2)-matrices. INTERNAL MODEL CONTROL DESIGN As presented before, the most popular structure for time delay compensation is the Smith Predictor struc- ture. A very important step in the multivariable Smith Predictor structure is the decoupling of the process. A modified multivariable Smith Predictor structure was proposed by Wang, Zou and Zhang (Wang et al., 2000). In this case the decoupling matrix is com- puted in frequency domain. More recently the Internal Model Control (IMC) method was used in order to compute the decoupling matrix (Wang et al., 2002). Chen et al. (2011) used the IMC method only as an intermediary step in order to compute the final con- troller – a matrix of PI controllers. The method used in this paper is based on the approach proposed by Pop et al. (2011) in which the IMC controllers are used as final controllers. The next step is to approximate the elements on the first diagonal of the matrix ( ) D H s with simple transfer functions: * ( ) ( )  iid iid Hf s Hf s (37) (37) The elements on the first diagonal, approximated by the transfer function given in (37), will be used for controller design. * The approximation of each element ( ) iid Hf s can be obtained by using genetic algorithms or graphical identification methods (Chen et al., 2011). MULTIVARIABLE SMITH PREDICTOR PID CONTROLLER DESIGN C sT f P e H H    Hf sT e ysp + - - + + + + + y d Figure 6: Smith predictor structure. Figure 7: Closed loop control scheme using the Smith predictor structure. All the controllers are computed using the imposed phase margin design method, for 60 . k    Figure 6: Smith predictor structure. The obtained controllers are: Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 162 Roxana Both, Eva-Henrie 11 12 21 22 1 ( ) 1.3259 1 ; 102.3018 1 ( ) 2.7733 1 ; 106.6667 1 ( ) 595.6621 1 ; 90.4977 1 ( ) 223.8721 1 ; 119.0476                                     HR s s HR s s HR s s HR s s (32) Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 162 # 1 (0) ( (0) (0) )    H H m m m m H H H H (35) (35) where # m H is the pseudo-inverse which will act as a pre-compensator matrix. pre-compensator matrix. (32) The decoupled multivariable process is done by: 11 12 # 21 22 ( ) ( )    d d D m m d d Hf Hf H s H s H Hf Hf (36) (36) The matrix (36) represents the steady-state decou- pled process, in which all elements are weighted sums of the original transfer functions . s ij Hf e    Due to the static decoupling, the steady-state matrix ( ) 0  D H s will be equal to the unit matrix. In this way all the elements which are not on the first diago- nal will be equal to zero. INTERNAL MODEL CONTROL DESIGN The model of the process is assumed to be equal to the process transfer function matrix Hp, presented before: The next step is to design the IMC controller as follows (Chen et al., 2011): 11 12 21 22 ( ) ( ) ( ) ( ) ( )          qo ti qoc tic s s m m m s s m m Hf s e Hf s e H s Hf s e Hf s e     (33) 11 12 21 22 ( ) ( ) ( ) ( ) ( )          qo ti qoc tic s s m m m s s m m Hf s e Hf s e H s Hf s e Hf s e     (33) * ( ) ( ) ( )    IMCi iid inv HR s Hf s f s (38) (38) (33) where ( ) f s is the IMC filter and ( )  iid inv Hf s repre- sents the invertible part of ( ) iid Hf s . The IMC filter is computed as: The next steps in order to design the IMC control- lers are: a) Determine the pseudo-inverse matrix; b) Determine the decoupled process; c) Approximate the elements on the first diagonal of the steady-state de- coupled process matrix; d) Design the IMC controllers. 1 ( ) ( 1)   n i f s s  (39) (39) The method used to decouple the multivariable system is the pseudo-inverse (Skogestad, 2011) of the steady state gain matrix: where λi is the time constant of the filter associated with each output and n is chosen such that the final IMC controller is proper. The parameters of the IMC controller will be tuned in accordance with the im- posed performance values of overshoot and settling time. Figure 8 and Figure 9 show the closed loop response of 11d Hf and 22d Hf considering different values of the IMC filter, λ = 5, 10, 15, 20. Brazilian Journal of Chemical Engineering SIIMULATION RESULTS 500 3000 λ=5 λ=10 λ=15 λ=20 40) 0 H H 1 2 100 4 20 1 IMC IMC HR HR   2 2 2 2 00 55.85 400 40 0001 87.2 100 20 s s s s s s       1 and 1 2 1 1 s s     (40) (40) 1000 440 442 444 446 448 450 452 Tout [K] 1500 T 2000 250 Time [s] 500 3000 λ=5 λ=10 λ=15 λ=20 0 the con The first sim e set-point tr ntrol strategi mulation sce racking perf ies. enario is focu formances of used on testin f the designe ng ed the con ou enc tem To this end e output tem nsidered. Fig utput tempera ce of the re mperature on a reference mperature of gure 11 pres ature while F eference step n the 2-ethyl- step variatio f the produc sents the ev Figure 12 sh p variation f -hexanol con on of 10 K f ct at t=10s volution of th hows the infl for the outp ncentration. for is he lu- put Figure 8: Tuning of the controller IMC1. Figure 1000 5.35 5.4 5.45 5.5 5.55 5.6 5.65 Coct [Kmol/m3] 9: Tuning of 1500 T f the controll 2000 250 Time [s] ler IMC2. 500 3000 λ=5 λ=10 λ=15 λ=20 0 Fig var gure 11: Ou riation on inp 0 1000 440 442 444 446 448 450 452 Tout [K] utput temper put 1. 00 2000 30 Tim rature evolut 3000 4000 Time [s] IMC PID tion for a ste 5000 6000 C D ep Figure 9: Tuning of the controller IMC2. Fig var gure 11: Ou riation on inp utput temper put 1. rature evolution for a ste ep u M co an th tr fu Figure 10 sing delay Moore-Penros ompensator f nd the fast m he time delay rix of all elem unction matri presents the compensator se pseudo-inv for the proces model ( ) m H s ys).  * D H s ments ( iid Hf ix ( ) D H s w closed loop r and IMC verse # m H is ss ( ) p H s , the ) (the process is the transf ( )s and * ( D H s without the tim control sche controller. INTERNAL MODEL CONTROL DESIGN Analyzing the simulation results in terms of settling time and 11 0 12 0 21 0 22 0 (0)  m m m m m Hf Hf H Hf Hf (34) (34) The pseudo-inverse is computed based on the fol- lowing equation (Skogestad, 2011): Brazilian Journal of Chemical Engineering 1163 Advanced Control o of a Complex Chemical Process Brazilian Journal of Chemical Engineerin ng Vol. 33, No. 001, pp. 155 - 1688, January - March, 2016 SIIMULATION RESULTS o st vershoot, th tudy results i he final IMC in the follow C controller wing transfer rs for the c function: case H H 1 2 100 4 20 1 IMC IMC HR HR   Figure Figure 1000 440 442 444 446 448 450 452 Tout [K] 1000 5.35 5.4 5.45 5.5 5.55 5.6 5.65 Coct [Kmol/m3] 2 2 2 2 00 55.85 400 40 0001 87.2 100 20 s s s s s s       8: Tuning of 9: Tuning of 1500 T 1500 T 1 and 1 2 1 1 s s     f the controll f the controll 2000 250 Time [s] 2000 250 Time [s] (4 ler IMC1. ler IMC2. 500 3000 λ=5 λ=10 λ=15 λ=20 500 3000 λ=5 λ=10 λ=15 λ=20 40) 0 0 im Gu in con sce tur bo ba sar con Th sce For analysis mplemented i uide, 2008). T comparison nclude the re enarios are t rbance reject th control st sed on the li ry to test th nsidering th his analysis enario. s purposes b in Matlab/S The simulati for the two c esults. The t the set-point tion analysis trategies, the inear model he robustness he variation is performe oth control s SIMULINK ion scenarios control strateg two main obj tracking ana s. Due to the controllers w of the proce s of the con of the mode ed in the thi structures we (Matlab Us s are presente gies in order jectives of th alysis and di e fact that, f were designe ess, it is nece ntrol strategi el parameter ird simulatio ere ser ed to he is- for ed es- ies rs. on H H 1 2 100 4 20 1 IMC IMC HR HR   Figure 1000 440 442 444 446 448 450 452 Tout [K] 2 2 2 2 00 55.85 400 40 0001 87.2 100 20 s s s s s s       8: Tuning of 1500 T 1 and 1 2 1 1 s s     f the controll 2000 250 Time [s] (4 ler IMC1. SIIMULATION RESULTS T s used as a p e model m H s model with fer function m )s is the trans me delay. eme The pre- ( )s hout ma- sfer Coct [Kmol/m3] 0 1000 5.29 5.3 5.31 5.32 5.33 5.34 5.35 5.36 Coct [Kmol/m3] 00 2000 30 Tim 3000 4000 ime [s] IM P 5000 6000 IMC PID Fig for Coct [Kmol/m3] gure 12: 2-E r a step varia 0 1000 5.29 5.3 5.31 5.32 5.33 5.34 5.35 5.36 Coct [Kmol/m3] Ethyl-hexano ation on inpu 00 2000 30 Tim ol concentra ut 1. 3000 4000 ime [s] IM P ation evolutio 5000 6000 IMC PID on F co Figure 10: C ontrollers. Closed loop control scheme using IM MC Fig for gure 12: 2-E r a step varia Ethyl-hexano ation on inpu ol concentra ut 1. ation evolutio on F co Figure 10: C ontrollers. Closed loop control scheme using IM MC 0.2 For the sam 25 [kmol/m3] me analysis a ] is consider reference ste red for the 2- ep variation -ethyl-hexan of nol Brazilian Journal of Chemical Engineerin ng Vol. 33, No. 001, pp. 155 - 1688, January - March, 2016 Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 164 output concentration. The evolution of the 2-ethyl- hexanol concentration is presented in Figure 13. and the influence of the reference step variation on the output temperature is detailed in Figure 14. Figure 16: 2-Ethyl-hexanol concentration evolution for a disturbance in 2-ethyl-hexenal flow. 0 1000 2000 3000 4000 5000 6000 5.3 5.31 5.32 5.33 5.34 5.35 5.36 5.37 5.38 5.39 5.4 Time [s] Coct [Kmol/m3] IMC PID To reach the second objective of the section, the second simulation scenario is focused on the disturb- ance rejection analysis of the designed control strate- gies. Figure 15 and Figure 16 present the effects of a 0.25 [kmol/m3] disturbance in 2-ethyl-hexenal flow on the two considered outputs. Figure 16: 2-Ethyl-hexanol concentration evolution for a disturbance in 2-ethyl-hexenal flow. Figure 13: Output temperature evolution for a step variation on input 2. SIIMULATION RESULTS 0 1000 2000 3000 4000 5000 6000 438 440 442 444 446 448 450 452 Time [s] Tout [K] IMC PID 0 1000 2000 3000 4000 5000 6000 438 440 442 444 446 448 450 452 Time [s] Tout [K] IMC PID From all the above figures it can be concluded that the IMC control strategy presents better perfor- mances in comparison with the MIMO PID control. Another advantage of the method is that it is more straightforward and easy to design. The minor disad- vantage is the Lag-Lead form of the controllers op- posite to the classical PID forms. Another advantage of the method is that it is more straightforward and easy to design. The minor disad- vantage is the Lag-Lead form of the controllers op- posite to the classical PID forms. The last objective of this section is to test the ro- bustness of the designed control strategies. To this end, the following figures present the evolution of the output temperature and 2-ethyl-hexanol concen- tration considering a reference step variation for both variables and also the variation of the uncertain pa- rameters of the linear plant. Figure 13: Output temperature evolution for a step variation on input 2. Figure 14: 2-Ethyl-hexanol concentration evolution for a step variation on input 2. 0 1000 2000 3000 4000 5000 6000 5.35 5.4 5.45 5.5 5.55 5.6 5.65 Time [s] Coct [Kmol/m3] IMC PID 0 1000 2000 3000 4000 5000 6000 5.35 5.4 5.45 5.5 5.55 5.6 5.65 Time [s] Coct [Kmol/m3] IMC PID The closed loop simulations using the multivaria- ble Smith Predictor PI controller under nominal pa- rameter values (solid black line), as well as the simu- lations considering the variation of the uncertain parameters of the linear plant (solid grey lines), for a reference step variation of the output temperature are presented in Figure 17 and Figure 18. The uncertain parameters considered are the time constants of the process as well as the gain parameter. Figure 14: 2-Ethyl-hexanol concentration evolution for a step variation on input 2. Figure 17: Output temperature evolution for a step variation on input 1– nominal case vs. uncertain case (PID). SIIMULATION RESULTS 1000 2000 3000 4000 5000 6000 440 442 444 446 448 450 452 454 Time [s] Tout [K] 1000 2000 3000 4000 5000 6000 440 442 444 446 448 450 452 454 Time [s] Tout [K] Brazilian Journal of Chemical Engineering Figure 15: Output temperature evolution for a dis- turbance in 2-ethyl-hexenal flow. Figure 17: Output temperature evolution for a step variation on input 1– nominal case vs. uncertain case (PID). 0 1000 2000 3000 4000 5000 6000 440 441 442 443 444 445 446 447 Time [s] Tout [K] IMC PID 1000 2000 3000 4000 5000 6000 440 442 444 446 448 450 452 454 Time [s] Tout [K] Figure 15: Output temperature evolution for a dis- turbance in 2-ethyl-hexenal flow. 0 1000 2000 3000 4000 5000 6000 440 441 442 443 444 445 446 447 Time [s] Tout [K] IMC PID 0 1000 2000 3000 4000 5000 6000 440 441 442 443 444 445 446 447 Time [s] Tout [K] IMC PID Figure 17: Output temperature evolution for a step variation on input 1– nominal case vs. uncertain case (PID). Figure 15: Output temperature evolution for a dis- turbance in 2-ethyl-hexenal flow. Brazilian Journal of Chemical Engineering Advanced Control of a Complex Chemical Process 165 In order to choose the control algorithm with the best performances, the same simulation scenario is considered for the IMC controller. To this end, the closed loop simulations using the IMC controller in Smith Predictor structure under nominal parameter values (solid black line), as well as the simulations considering the variation of the uncertain parameters of the linear plant (solid grey lines), for a reference step variation of the output temperature are presented in Figure 21 and Figure 22. Figure 18: 2-Ethyl-hexanol concentration evolution for a step variation on input 1– nominal case vs. un- certain case (PID). 1000 2000 3000 4000 5000 6000 5.2 5.25 5.3 5.35 5.4 5.45 Time [s] Coct [Kmol/m3] Figure 21: Output temperature evolution for a step variation on input 1– nominal case vs. uncertain case (IMC). 1000 2000 3000 4000 5000 6000 440 442 444 446 448 450 452 454 Time [s] Tout [K] 1000 2000 3000 4000 5000 6000 440 442 444 446 448 450 452 454 Time [s] Tout [K] Figure 18: 2-Ethyl-hexanol concentration evolution for a step variation on input 1– nominal case vs. un- certain case (PID). SIIMULATION RESULTS The closed loop simulations using the multivaria- ble Smith Predictor PI controller under nominal pa- rameter values (solid black line), as well as the simu- lations considering the variation of the uncertain parameters of the linear plant (solid grey lines), for a reference step variation of the 2-ethyl-hexanol con- centration are presented in Figure 19 and Figure 20. Figure 21: Output temperature evolution for a step variation on input 1– nominal case vs. uncertain case (IMC). Figure 19: Output temperature evolution for a step variation on input 2– nominal case vs. uncertain case (PID). 1000 2000 3000 4000 5000 6000 430 435 440 445 450 455 Time [s] Tout [K] 1000 2000 3000 4000 5000 6000 430 435 440 445 450 455 Time [s] Tout [K] Figure 22: 2-Ethyl-hexanol concentration evolution for a step variation on input 1– nominal case vs. un- certain case (IMC). 1000 2000 3000 4000 5000 6000 5.3 5.31 5.32 5.33 5.34 5.35 5.36 5.37 Time [s] Coct [Kmol/m3] 1000 2000 3000 4000 5000 6000 5.3 5.31 5.32 5.33 5.34 5.35 5.36 5.37 Time [s] Coct [Kmol/m3] Figure 19: Output temperature evolution for a step variation on input 2– nominal case vs. uncertain case (PID). Figure 22: 2-Ethyl-hexanol concentration evolution for a step variation on input 1– nominal case vs. un- certain case (IMC). Brazilian Journal of Chemical Engineering Vol. Figure 20: 2-Ethyl-hexanol concentration evolution for a step variation on input 2– nominal case vs. un- certain case (PID). 1000 2000 3000 4000 5000 6000 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 Time [s] Coct [Kmol/m3] 1000 2000 3000 4000 5000 6000 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 Time [s] Coct [Kmol/m3] The closed loop simulations using the IMC con- troller in Smith Predictor structure under nominal parameter values (solid black line), as well as the simulations considering the variation of the uncertain parameters of the linear plant (solid grey lines), for a reference step variation of the 2-ethyl-hexanol con- centration are presented in Figure 23 and Figure 24. Based on the simulation results presented, it can be concluded that the most appropriate control strat- egy with satisfactory results is the IMC control strat- egy in Smith Predictor structure. Figure 20: 2-Ethyl-hexanol concentration evolution for a step variation on input 2– nominal case vs. un- certain case (PID). Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 NOMENCLATURE Ci molar concentration (kmol/m3) cpi specific heat capacity (kJ/kmol K) F volumetric flow rate (m3/s) T Temperature (K) ΔRH reaction heat (kJ/kmol) K adsorption equilibrium constant, component i (m3/mol) k rate constant of surface reaction j (mol/s kg) HH2 Henry’s law constant v velocity (m/s) S reactor cross-sectional area (m2) M molecular mass (kg/kmol) av specific gas-liquid contact area (m-1) r rate of surface reaction (mol/s kg) t time (s) z axial reactor coordinate Greek Letters ρ density (kg/m3) Subscripts/Superscripts G gas phase L liquid phase i H2, 2-ethyl-hexenal, 2-ethyl-hexanal, 2-ethyl-hexanol j 2-ethyl-hexenal hydrogenation reaction, 2-ethyl-hexanal hydrogenation reactor Ci molar concentration (kmol/m3) cpi specific heat capacity (kJ/kmol K) F volumetric flow rate (m3/s) T Temperature (K) ΔRH reaction heat (kJ/kmol) K adsorption equilibrium constant, componen i (m3/mol) k rate constant of surface reaction j (mol/s kg HH2 Henry’s law constant v velocity (m/s) S reactor cross-sectional area (m2) M molecular mass (kg/kmol) av specific gas-liquid contact area (m-1) r rate of surface reaction (mol/s kg) t time (s) z axial reactor coordinate Greek Letters ρ density (kg/m3) Subscripts/Superscripts G gas phase L liquid phase i H2, 2-ethyl-hexenal, 2-ethyl-hexanal, 2-ethyl-hexanol j 2-ethyl-hexenal hydrogenation reaction, 2-ethyl-hexanal hydrogenation reactor Figure 23: Output temperature evolution for a step variation on input 2– nominal case vs. uncertain case (IMC). 0 1000 2000 3000 4000 5000 6000 420 425 430 435 440 445 Time [s] Tout [K] 0 1000 2000 3000 4000 5000 6000 420 425 430 435 440 445 Time [s] Tout [K] Figure 23: Output temperature evolution for a step variation on input 2– nominal case vs. uncertain case (IMC). Figure 24: 2-ethyl-hexanol concentration evolution for a step variation on input 2– nominal case vs. un- certain case (IMC). 1000 2000 3000 4000 5000 6000 5 5.2 5.4 5.6 5.8 6 6.2 Time [s] Coct[Kmol/m3] 1000 2000 3000 4000 5000 6000 5 5.2 5.4 5.6 5.8 6 6.2 Time [s] Coct[Kmol/m3] Figure 24: 2-ethyl-hexanol concentration evolution for a step variation on input 2– nominal case vs. un- certain case (IMC). CONCLUSIONS This work was supported by a grant of the Roma- nian National Authority for Scientific Research, CNDI– UEFISCDI, project number 155/2012 PN-II- PT-PCCA-2011-3.2-0591. This paper describes the two control strategies of an important chemical process, the 2-ethyl-hexenal hydrogenation process, in a catalytic fixed-bed reac- tor. This process is a nonlinear system with dis- tributed parameters. However, it is possible to de- scribe the processes that occur inside the reactor by a linear nominal transfer matrix. SIIMULATION RESULTS It can be observed that the differences between the performances of the nominal and perturbed sys- Brazilian Journal of Chemical Engineering Vol. 33, No. 01, pp. 155 - 168, January - March, 2016 Roxana Both, Eva-Henrietta Dulf and Ana-Maria Cormos 166 tems are smaller than in the case of classical control, Figure 17 to Figure 24. It must be mentioned that the simulation scenarios are considered for four extreme points of operation, implying a very large range of parameter variation. If the variation range of the uncertain parameters is smaller, in normal operation conditions, the differences between the performances of the nominal and perturbed systems are very small. IMC control presents better performances for set- point tracking and also for disturbance rejection tests, and better robustness, so it is recommended for implementation. Additionally, this method reduces the design burden of the controllers. 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https://openalex.org/W4312137522	https://revistafenix.bnp.gob.pe/index.php/fenix/article/download/620/2089	es		Lima estrellada	Fénix/Fénix	2,022	cc-by	7,007	Matías Maggio-Ramírez ISSN-e: 2709-5649 pp. 110-125 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires Starry Lima. A reading of the works of Pedro Peralta Barnuevo and Hipólito Unanue in the foundation of the Public Library of Buenos Aires Matías Maggio-Ramírez Universidad Nacional de Tres de Febrero Buenos Aires, Argentina Contacto: mmramirez@untref.edu.ar https://orcid.org/0000-0002-1106-1381 Resumen El objetivo del artículo es analizar la pervivencia de la figura del cielo estrellado en dos obras: Lima fundada, de José Peralta Barnuevo, y Observaciones sobre el clima de Lima y sus influencias en los seres organizados, en especial el hombre, de Hipólito Unanue, desde la historia cultural. La escritura poética y metafórica, a pesar de que estos libros trataran cuestiones propias del género ensayístico y pusieran de relieve la salubridad del clima limeño, permitió indagar en distintas tradiciones sobre la representación del cielo estrellado en la estructura del poema épico y en la influencia de los astros en las personas. Palabras clave: cielo estrellado, Lima, determinismo climático, historia natural, poesía épica. Abstract The aim of this article is to analyse the survival of the figure of the starry sky in two works, Lima fundada by José Peralta Barnuevo and Observaciones sobre el clima de Lima y sus influencias en los seres organizados, en especial el hombre by Hipólito Unanue, from cultural history. Poetic and metaphorical writing, even though books dealt with issues typical of the essay genre and highlighted the salubrity of Lima´s climate made it possible to inquire into different traditions about the representation of the starry sky in the structure of the epic poem and in the influence of the stars in people. Keywords: starry sky, Lima, climatic determinism, natural history, epic poetry. Recibido: 2022-06-03/ Revisado: 2022-09-23 / Aceptado: 2022-09-24/ Publicado: 2022-12-06 110 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires Esta peculiaridad de las inteligencias bien dotadas ha sido representada en el Perú por dos hombres que, durante dos siglos, y cada uno dominando en el suyo, se dividieron el imperio de la sabiduría. Peralta lleva el cetro durante el último tercio del siglo XVII y los primeros períodos del XVIII; Unanue lo hereda de manos de aquél y lo sostiene sin rival, pero rodeado de discípulos, hasta el año 1833 en que falleció octogenario. Juan María Gutiérrez (1957, p. 16) 1. Lecturas bonaerenses La fundación de la Biblioteca Pública de Buenos Aires fue una de las instancias centrales para comprender la cultura impresa del periodo virreinal en el Río de la Plata. Con ese gesto, la Primera Junta de Gobierno favoreció «la conservación de libros y la lectura de estudio como vehículo de aprendizaje social» (Parada, 2009, p. 174), a la vez que fue una prueba del proceso revolucionario que visibilizó un espectro de lecturas e intereses que se creyó necesario sociabilizar y compartir. El texto fundacional, publicado en la Gazeta de Buenos Ayres el 13 de setiembre de 1810, reconocía aliviado que «por fortuna tenemos libros bastantes para dar principio a una obra, que crecerá en proporción del sucesivo engrandecimiento de este pueblo», por lo que se abrió una suscripción patriótica para los gastos de estantes y demás costos «inevitables». En el libro de ingresos y donaciones, así como en la Gazeta, se dejó noticia de la participación de la comunidad que entregó libros, periódicos, maderas, cuadernos y dinero para acompañar la iniciativa institucional de la Primera Junta1. La creación de la Biblioteca Pública de Buenos Aires fue la apropiación de un gesto del regalismo propio de las reformas borbónicas. La expulsión de los jesuitas había dejado una gran biblioteca, aunque en parte fue vendida a particulares. Antes de decretarse la fundación de la Biblioteca ya se había solicitado el envío de los libros que quedaran en los distintos edificios de los jesuitas para incorporarlos en el catálogo fundacional. La importante presencia de libros en lenguas vernáculas como el castellano y el francés dejó entrever que la Biblioteca crecería más allá de los límites religiosos donde reinaba el latín. El saber laico buscaba la clave para lograr la felicidad pública como sinónimo del progreso en libros y periódicos. 1 En el libro de donaciones se registró el autor, el título de la obra en castellano —aunque la obra se encontrara en otro idioma salvo en latín—, la cantidad de tomos y el formato del libro. Se han realizado dos transcripciones parciales del libro de donaciones en La Revista de la Biblioteca Pública de Buenos Aires, bajo la dirección de Manuel Ricardo Trelles en 1879, en este caso con la transcripción de las noticias sobre la Biblioteca en la Gazeta de Buenos Ayres; y en la Revista de la Biblioteca Nacional, dirigida por Felipe Barreda Laos en 1944. Para rastrear los títulos que aquí nos interesan se tuvieron en cuenta tanto las noticias sobre la Biblioteca en la prensa como las transcripciones del libro de donaciones, así como su cotejo con fotografías digitales del libro manuscrito. Los registros del libro de donaciones permiten analizar el horizonte de expectativas y temas que interesaban a los lectores que fueron los principales donantes. 111 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez Entre los poco más de mil libros que fueron registrados entre 1810 y 1824, dos títulos de autores peruanos se encontraron en el libro de ingresos y donaciones, los cuales mantenían un diálogo entre ellos2. Entre los ejemplares extraídos del Colegio San Carlos y algunos de los que eran de su rector Luis José de Chorroarín, en 1810 se registró la entrada: «Peralta. El poema histórico titulado: Lima fundada, con las notas históricas declaratorias &c. en p.no» en dos volúmenes. En 1822 se anotó en el libro que «El D.r D.n Hipolito Vnanue, presentó su obra intitulada Observaciones sobre el clima de Lima y sus influencias en los seres organizados, 2.da edicion Madrid 1815, 4° p.ta» en un volumen (Primeras donaciones…, 1944)3. Ambas obras utilizaron la fuerza de la escritura poética para argumentar tanto sobre la importancia del clima en la salud de los limeños como para narrar la historia de una ciudad, sin perder de vista el cielo estrellado. 2. Hipólito Unanue La constitución del cielo influye en los seres organizados que habitan debaxo de él. Pende de aquí especialmente el tono de la naturaleza productiva, y la calidad de sus partos. Por esto en los tiempos antiguos en que florecieron las ciencias, el estudio de los climas fue uno de los que mejor se cultivaron. Hipólito Unanue (1815) Unos meses antes del temblor que en 1806 sacudió las torres de Lima, se publicó en aquella ciudad la primera edición de Observaciones sobre el clima de Lima y sus influencias en los seres organizados, en especial el hombre. Su autor, Hipólito Unanue, catedrático de Medicina en la Real Universidad de San Marcos, gozó de fama y reconocimiento al ser su obra apreciada «no sólo por Literatos de América, sino también de los de Europa», como Alexander von Humboldt en su Ensayo político sobre la Nueva España4. La segunda edición corregida por el autor se entregó a imprenta en Madrid luego de que, en octubre de 1807, se viera en Lima un cometa y en noviembre de 1811, otro que permaneció visible 2 Se recortó el periodo entre esas fechas porque entre 1825 y 1833 no se registró ningún tipo de material en el libro de ingresos. Se privilegió el relevamiento de esa fuente institucional, antes que la periodística, como signo del pasaje de las bibliotecas privadas a las públicas. Por otro lado, no son los únicos autores o temas peruanos que aparecen en el catálogo, pero en este primer acercamiento al análisis de la fuente se hizo esta selección por la cita de Unanue a Peralta 3 Lamentablemente, ninguno de los títulos donados se encuentra en el Tesoro de la Biblioteca Nacional Mariano Moreno. El ejemplar de Unanue de 1815 pertenece a la donación Balcarce. 4 Agradezco la lectura de José Emilio Burucúa al primer borrador de este trabajo que se presentó en su seminario «Tradición clásica y mundo moderno» que dictó en el Instituto de Altos Estudios Sociales de la Universidad Nacional de San Martín. 112 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires durante seis meses sin necesidad de telescopio. Unos años más tarde, un gran incendio en Guayaquil destruyó media ciudad, un huracán arrancó de raíz varios árboles de la alameda de Lima y hubo terremotos en Ica y Piura (Palma, 1993, p. 118). El 2 de abril de 1814, las aguas oscuras del tintero de Unanue descansaron. Se envió el manuscrito a Madrid, donde se imprimió con las licencias necesarias la nueva edición al año siguiente, mientras que, en Lima, por esos tiempos de clima independentista, el virrey José Fernando de Abascal recibía en su escritorio —no con sorpresa—tres pequeñas bolsas con sal, habas y cal. En la edición de 1815 de la obra de Unanue se citaba, en la Advertencia, el juicio crítico del Memorial Literario de Madrid5 a la primera edición donde, tras indicar: […] lo versado que esta [Unanue] en las ciencias naturales, y también que no le son extrañas las humanidades; pero lo que se advierte con más particularidad es el caudal de buenos conocimientos anatómicos y médicos de que está adornado, y la mucha erudición con particularidad de los autores ingleses. Es preciso confesar no obstante que el castellano es incorrecto, y que suele a veces el autor exaltar su imaginación de tal modo que en las narraciones emplea el estilo propio de las descripciones poéticas […]. (Unanue, 1815, p. 6). Ante el juicio del lector español —que lee en la prosa del médico un exceso de imaginación por donde se cuela la escritura poética—, Unanue redobló la apuesta. Esgrimió metáforas para explicar que la imaginación acude como el calor de primavera que viste de hojas y flores los árboles desnudos del invierno. Sabía que no había hecho más que «[…] interpon[er] en la narración filosófica las imágenes y descripciones poéticas, teniendo por maestro a Platón, quien, como observa Longino, lo executa con freqüencia en sus tratados filosóficos» (Unanue, 1815, p. 7). La clásica tradición escolástica consideraba a la poesía como «“infima inter omnes doctrinas” (Summa theologica, I, i,9)» (Rico, 1997, p. 32). El mismo enfoque también se encontraba en San Alberto, al afirmar «poeticus modus infirmior est inter modos philosophiae», ya que la escolástica no se interesó por la dignificación de la poesía (Curtius, 2017, p. 318). Unanue, ilustrado y neoclásico, citó en su ensayo los versos de Horacio y Virgilio. Hasta un fragmento de la obra épica de Torcuato Tasso Jerusalén liberada es glosado por Unanue en sus Observacio5 Se supone que Unanue se refiere al periódico Memorial literario instructivo y curioso de la Corte de Madrid que con periodicidad mensual primero y, luego bimensual, se publicó en varias etapas: «desde 1784 a 1791, de 1793 a 1797, y de 1801 a 1808». Cada número tenía alrededor de 125 páginas en octavo y costaba de 3 a 4 reales. Su fundador fue Joaquín Ezquerra y contaba con el auxilio de Pedro P. Trullech (Sáinz, 1983, p. 186). 113 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez nes. El gesto neoclásico del médico buscaba insertar su obra en una antigua tradición para validar su propia escritura, al igual que el historiador y poeta del siglo XIV Albertino Mussato. Este sostenía que el saber necesita y utiliza la poesía, ya que no se puede prescindir de Virgilio, Ennio y Homero porque: Con ellos corroboran los filósofos sus dichos, y con ellos los juristas, los gramáticos, rétores, dialécticos, los que inquieren del mundo los secretos: sin nuestra Musa no hay saber ni técnica; y aun a la propia Iglesia, no lo olvides, oirás cantar también con nuestra métrica. (Rico, 1997, p. 32). Unanue encontró la forma de articular la poesía como recurso estilístico y cita de autoridad para el saber científico e ilustrado. Su escritura encaraba los límites del ensayo medicinal y sorprendía a los lectores españoles al cruzar el umbral de la portadilla. En la América hispánica, y principalmente en el Perú, el carácter atrabiliario aquejó no solo a los pueblos originarios sino también a los españoles criollos. Las lecturas inglesas de Unanue se revelan cuando sostiene que el hombre adquiere todos sus conocimientos por medio de las imágenes exteriores que transmiten los sentidos, por lo que ante un cielo despejado y brillante […] harán que desde la niñez se imprima el cuño de las gracias festivas, y su expresión la risa. Un cielo nebuloso ofreciendo las ideas con un ayre opaco, introducirá las semillas de los sentimientos melancólicos; mucho más si la imaginación es sensible, y delicados los nervios: porque entonces las impresiones se hacen completas y retienen con tenacidad (Unanue, 1815, p. 134). Este último temperamento, indicó más adelante, era el que regía a los aborígenes del Perú. La melancolía afectó, según Unanue, a varias naciones que llegaron a Lima porque «los cuerpos han cedido o contrariado a las impresiones debilitativas del clima» (185, p. 137)6. El estilo poético de Unanue sería entonces propio de los habitantes de Lima, síntoma de la influencia del clima, de las estaciones y del cielo. Desde el título, la obra se insertó en la tradición de la medicina ligada al determinismo climático, que sostenía que los climas cálidos fomentaban la 6 En 1683, Francisco Montaldo fue el canónigo encargado de la defensa en el proceso de beatificación de fray Toribio de Mogrovejo. En el panegírico comienza con una descripción de la ciudad de Lima en la cual la relaciona con Saturno: «[…L]a ciudad de Lima […] Tiene por estrellas verticales tres famosos astros, que con sus generosas influencias, y con la templanza que resulta de su propia oposición, fecundan.[…] Melancólicos influjos causarán sin duda estas estrellas verticales a no suprimirse la térrea calidad de Saturno con la benéfica de Mercurio, que reprime sus daños, y causa templadísimos efectos» (Sala Catalá,1994, p. 283). 114 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires pereza y la melancolía, pero, a diferencia de los eruditos europeos, creía que había factores morales que podían hacer contrapeso. El macrocosmos dejaría su huella en la escritura, florearía los textos científicos y también influiría en la música. A quienes deseaban disipar la melancolía se les recomendaba buscar la conversación de un amigo o escuchar música porque esta podía «refrenar los delirios» (Unanue, 1815, p. 240). El cielo y las estrellas tenían, aún en el siglo XIX, una relación con el mundo natural, al determinar la salud. La Iglesia, según el Compendio Moral Salmaticense de Marcos de Santa Teresa, impreso 1805, apoyaba la astrología natural porque [...] es cuando por la diversa disposición de los astros, por su situación, movimiento, resplandor, nacimiento, y ocaso se anuncia agua, frío, calor, o eclipse, u otros efectos naturales; y ésta es lícita, según opinión común; porque los dichos efectos pueden naturalmente conocerse por sus causas. En cambio, la astrología judiciaria o genetlíaca «es, cuando del curso de los astros, de su conjunción, y aspecto se anuncian los efectos futuros fortuitos, o los presentes y pasados del todo ocultos, y las acciones libres de los hombres»7. 7 En 1726, Benito Feijoo, en su Teatro crítico universal, ataca a la astrología judiciaria y a los almanaques (Tomo I, Discurso VIII). Los argumentos que desgrana en 44 puntos contra los genetlíacos bien pudieron ser inspirados, además de los clásicos dominicos Santo Tomas y San Agustín, por los textos que J. Swift publicó en febrero de 1708 bajo el seudónimo de Isaac Bickerstaff. Addison y Steele, amigos de Swift, encarnaron a Bickerstaff en la cruzada contra la astrología encabezada por John Hewson que firmaba con el ya famoso seudónimo John Partridge su almanaque judiciario Merlin. En 1710, los viejos colegas de The Spectator escribieron en The Tatler n° 216 que «Es verdad que John Partridge no sólo murió en aquel tiempo [28 de marzo de 1708] sino que continúa muerto al día de hoy [24 al 26 de agosto de 1710]». Los textos de The Tatler fueron prontamente recopilados en libros con el título The Lucubrations of Isaac Bickerstaff. Esta hipótesis apresurada y jocosa no podrá llegar a comprobarse, ya que luego de su muerte la biblioteca del padre Feijoo fue trasladada al monasterio de Samos y desapareció posteriormente en un incendio. Feijoo también cargará su pluma siempre afilada contra el astrólogo francés Juan Morin, quien fuera apreciado por el rey cristianísimo Luis XIII, en el tomo I, carta XXXVIII de sus Cartas eruditas y curiosas. Como curiosidad, en Buenos Aires, los dos tomos en in cuarto de The Lucubrations se encontraban en el inventario de la biblioteca de Juan Hipólito Vieytes. Para el año 1790, en la Imprenta de Don Antonio de Sacha, en Madrid, se imprimió el Índice último de los libros prohibidos y mandados a expurgar: para todos los reinos y señoríos… Allí aparecen los libros puestos en el Índice Expurgatorio desde el año 1747 hasta fines de diciembre de 1790 bajo el celoso cuidado del Inquisidor General D. Agustín Rubín de Ceballos. En la regla IX de las Reglas, mandatos y advertencias generales, página xx, puede leerse que de forma taxativa «[…] se prohíben los Libros, Tratados, Índices, Cédular, Memoriales, Recetas, Nóminas, Escritos y Papeles de Geomancia, o Hidromancia, Aeromancia, Piromancia, Onomancia, Chiromancia, Nigromancia, o en que se contienen sortilegios, hechizos, o cualesquier agüeros, encantaciones, De Arte Mágica, divinaciones, brujerías, cercos, caracteres, sellos, sortijas y figuras, o invocaciones de demonios, en qualquier manera que sea. Ítem, todos los de Astrología Judiciaria, que llaman de nacimientos, y levantar figuras, interrogaciones y elecciones, en que se afirman, o dan Reglas, o se enseña Arte o Ciencia para conocer por las Estrellas y sus aspectos futuros contingentes sucesos, o casos fortuitos o acciones que dependen de la voluntad libre humana, aunque en los tales 115 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez En la introducción de las Observaciones…, Unanue transcribió, luego de escribir sobre las bondades del sol en la costa limeña, «que hace disfrutar en casi todo el año un temple de Primavera» el canto VIII del poema épico Lima fundada, de Pedro Peralta y Barnuevo: En su horizonte el Sol todo es aurora Eterna el tiempo todo es Primavera; Solo es risa del Cielo cada hora, Cada mes solo es cuenta de la Esfera Son cada aliento un hálito de Flora, Cada arroyo una Musa lisonjera; Y los vergeles, que el confin le debe, Nubes fragantes con que al Cielo llueve. Para Unanue «el primer poeta de América ha cargado de coloridos el quadro de esta descripción, quando todos los viageros de Europa convienen, en ser el del Paraíso el temperamento de Lima» (1815, p. 20). Las palabras del polígrafo mercurista se anudan en un pasado, ya que apuntan a una de las claves de interpretación: el cielo estrellado ptolemaico, del poema Lima fundada. 3. Pedro Peralta y Barnuevo En 1732, desde la imprenta de Sobrino y Bados se da a conocer en la capital del virreinato del Perú el libro Lima fundada o conquista del Perú. Poema heroico en que se decanta toda la historia del descubrimiento y sujeción de sus provincias, por D. Francisco de Pizarro… escrito por el doctor Pedro de Peralta Barnuevo, Roca y Benavides. Al texto lo antecede una «Aprobación» del doctor Pedro José Bermúdez de la Torre y Solier, que también había sido rector de la Universidad de San Marcos. La «Aprobación», sostiene José Antonio Mazzotti, «es en sí misma el primer ejercicio crítico y analítico que se ofrece del largo poema desde el mismo momento de su publicación» (2006, p. 471). Mazzotti se adentra en el texto de Bermúdez para rastrear Libros y las demás cosas se diga y proteste que no se afirma de cierto, o en que se enseña a responder lo hecho o acontecido en las cosas pasadas, libres y ocultas de hurtos y otras semejantes, aunque se diga también que no se afirma de cierto. Y se prohíbe y manda que ninguna y manda que ninguna persona haga juicio acerca de las dichas cosas. Pero se permiten los juicios y naturales observaciones que están escritos, y se hacen para ayudar a la Navegación, Agricultura o Medicina […]». Dentro de la reglamentación se permite la circulación de textos ligados a la astrología natural porque «[…] proviene de causas naturales como los eclipses, lluvias, tiempos serenos o secos, pestes, los quales no pertenecen a divinación prohibida». La permeabilidad de la regla inquisitorial también aceptaba «los juicios de nacimientos, que sin afirmación enseñan a sospechar o conjeturar las inclinaciones, calidades, complexiones corporales de cada uno, sin pasar en manera alguna a los dichos futuros contingentes […] que penden de la voluntad libre». La astrología natural se encuentra ligada a los astros y a su influencia en el aquí y ahora o en la cadencia de su repetición como las estaciones del año o las mareas. 116 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires un ideal social de armonía y orden que […] muestra bastante bien las tensiones y pasiones subyacentes en el discurso criollista […] [a]simismo, la concepción poética y universalista de la «Aprobación» […] puede dar cuenta de un microcosmos cifrado en el poema de Peralta que sirve como programa de re-facción sociopolítica (p. 471). Allí encuentra que la presentación de Peralta como un Virgilio español, la alusión al erotismo del viajero y su recepción favorable en la tierra conquistada no son gratuitas, ni tampoco la reivindicación de los criollos como estrellas novomundiales. La estrategia de Bermúdez y Peralta es, siempre desde el prisma de Mazzotti, la conquista de un espacio discursivo propio para localizar la relación entre los descendientes de los conquistadores […] y las masas indígenas que eran constantemente presentadas en situación de inferioridad material y cultural. A la vez, los criollos desarrollaban la analogía para reclamar una posición de abandono injusto por parte de los «chapetones», de los que sin embargo no podía desligarse políticamente. (2006, p. 479). Fue necesaria, según el análisis que aquí se sigue, la asociación de la estructura narrativa —así como en su topoi, el cielo estrellado— con la tradición clásica en la que se demuestra un amplio bagaje de conocimientos sobre astronomía ptolemaica cuando había sido superada por Copérnico, Kepler y Galileo8. La estructura de los cielos en Lima fundada El poema está dividido en diez cantos que, siguiendo la clave interpretativa de Bermúdez desarrollada por Mazzotti, se corresponden con las nueve musas y Phebo (Apolo). Ellos, a su vez, se relacionaban con los «diez radiantes Olympicos espacios». La concordancia se sostiene de forma erudita al citar las «Tablas del rey Don Alfonso, seguidas de todos los Astrónomos modernos, que refiere el Padre Ricciolli en su Almagesto […] Según Macrobio y Glareano, que siguen a Hesyodo en su Theogonia». El esquema es el siguiente (Mazzotti, 2006, p. 481): 8 Francisco Ruiz Lozano comenzó a publicar en Lima, en 1656, un Repertorio de los Tiempos, en el cual —según José Sala Catalá (1994, p. 256)— se ofrecía al público las principales efemérides astronómicas y astrológicas del siguiente año en el que, por orden del virrey conde Alva de Liste, se funda para él una cátedra de Astrología y Matemáticas en la Universidad Mayor de San Marcos. Unos años después, en 1665, publicó su Tratado de Cometas, observación y juicio del que se vió en esta ciudad de los Reyes, y generalmente en todo el Mundo, por los fines del año 1664 y principios de 1665. Ya en 1650, Sala Catalá encuentra que se conocía la obra de Galileo, ya que el superintendente y gobernador de la mina de Huancavelica publicó una nota necrológica en la que se deja de lado el proceso inquisitorial y se «informa con mucho detalle de la originalidad de su obra». 117 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez Canto 1° Canto 2° Canto 3° Canto 4° Canto 5° Canto 6° Canto 7° Canto 8° Canto 9° Canto 10° Musa, Clío Musa, Euterpe Musa, Talía Musa, Melpómene Musa, Terpsícore Musa, Erato Musa, Polimnia Musa, Urania Musa, Calíope Canto, Phebo la Luna Mercurio Venus el Sol Marte Júpiter Saturno Cielo Sidéreo, o Estrellado Aqueo, o Cristalino el Primer Móvil Así como en el primer canto el tema es el viaje de Pizarro por mar bajo la mirada de la Luna que gobierna las mareas, el siguiente canto narra la embajada de Tito Atauchi dominado por Mercurio, patrono de los embajadores sabios, discretos y elocuentes, lo cual se encuentra en consonancia con el nombre de Euterpe, que significa «suavidad» y por esta razón inspira dulzura. En el canto tercero, «el tierno amante afecto que sacrificó, el mismo invicto Héroe a la hermosa Princesa, Hermana de Atahualpa» se encuentra regido por Venus, el orbe celeste y por la musa Talía. Así pues, el motivo de los cantos se encuentra enlazado con el cielo y con aquello que representan los dioses griegos y le da nombre a los cuerpos celestes en la tradición astrológica, la cual recupera Juan Bautista Riccioli en su Almagesto. Tal como sostiene Aby Warburg (2005, p. 417), «La astrología no es en el fondo sino un fetichismo onomástico proyectado al futuro»9. A fines de la Edad Media, las observaciones reales de las estrellas errantes y las fijas cedieron su puesto a un culto primitivo a los nombres de estos astros. La Edad Media heredó la astrología de la Antigüedad y la legó al Renacimiento para sobrevivir en el tiempo. «Los teólogos ortodoxos podían aceptar la teoría de que los planetas afectaran a los acontecimientos y a la psicología y, mucho más, a las plantas y los minerales. La iglesia no combatía esa idea» (Lewis, 1997, p. 85), pero sí, en cambio, cuestionaba la astrología judiciaria porque eliminaba el libre albedrío. El argumento principal era que el hombre justo podía vencer a las estrellas. La 9 Giorgio Agamben (2007, pp. 166-167), en su texto «Aby Warburg y la ciencia sin nombre», escribió que «[s]i Warburg pudo además presentar el problema del Nachleben des Heidentums [renacimiento del paganismo] como su propio problema supremo en tanto estudioso, fue porque comprendió, con una sorprendente intuición antropológica, que el de “la transmisión y la supervivencia” es el problema central de una sociedad “caliente” como la occidental, tan obsesionada por la historia como para querer hacer de ella el motor de su propio desarrollo. […] En esta perspectiva, donde la cultura es vista siempre como un proceso de Nachleben, es decir de transmisión, recepción y polarización, también se vuelve comprensible por qué Warburg debió fatalmente concentrar tanta atención sobre el problema de los símbolos como sobre su vida en la memoria social». 118 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires iglesia también luchó contra la adoración de los planetas. El poeta cristiano no creía en el dios porque creía en el planeta, sino en el cuerpo celeste, en su influencia sobre la tierra y en dios como una unidad en su mente. A pesar del amplio conocimiento de la ciencia astronómica con el que contaban los limeños en el siglo XVIII, tanto en la «Aprobación» como en el poema se retoman los ecos del Timeo de Platón, que serían fosilizados en la definición de astrología que en 1611 dio Sebastián de Covarrubias en su Tesoro de la lengua castellana como la «[c]iencia que trata del movimiento de los astros y los efectos que dellos proceden, cerca de las cosas inferiores y sus impresiones, que por otro nombre dicen astronomía». En Lima fundada, el orden celeste se repite. Ya no solo como se estableció en la «Aprobación», dando a los cantos la influencia en su contenido narrativo del «fetichismo onomástico» de los dioses paganos encarnados en los cuerpos celestes, sino en su interior. Siendo la tierra el elemento más pesado, se precipita en la mise en page como notas al pie. Desde allí, con su tipografía abigarrada y diminuta, es donde Peralta Barnuevo aparece para observar el movimiento de las esferas encarnadas en los cantos que narran la historia de la ciudad que nunca abandonó. Desde las notas, la astrología es natural, es propia de la observación en la que los nombres de los dioses paganos solo aparecen como glosa de un saber enciclopédico para dar sentido a la historia de Lima. También en la Tierra —es decir, en las notas al pie— se indican datos astronómicos-matemáticos, como en la nota n° 1 del canto VIII al verso De Acuario en los confines ilumina: «[p]asa por el zenit de Lima el grado 27, min. 48, seg. 8». En la nota siguiente puede leerse la explicación de las Espiras, Spiras o Hélices que «son líneas curvas semejantes a los tornillos, y estas son las que forma el Sol con su movimiento diurno, por la declinación que va teniendo cada día, aunque por lo insensible de la diferencia se llaman Círculos Paralelos, porque son a la Equinoccial». Pero es en la nota 3 donde aparece la primera persona, el observador del cielo que goza de reconocimiento internacional: Tiene Lima de latitud o altura de Polo austral 12 gr. 15 min. según mis nuevas observaciones, concuerdan con las del célebre Pedro Fernández de Quiroz. Su longitud es de 301 gr. 25 min. Puesto el primer meridiano en la isla del Ferro, asi mismo según mis observaciones, habiendo hallado la diferencia de meridianos entre esta ciudad y la de París de 5hor. 16 min. 20 seg. Con el cotejo de las hechas en Bolonia por Manfredi, y de las de la Real Academia de París, que por su carta me le participó. La cual halló M. Frezier tan verdadera, como lo expresa en la relación de su viage, donde me cita con testimonio individual. 119 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez Mientras la maquinaria de los cielos se encuentra en movimiento, su escritura no es azarosamente cantos repletos de versos de oralidad ficticia, tal vez con la vana intención de acompañar la música de las esferas. Entretanto, en las notas al pie, en el centro del universo, la escritura tiene la linealidad de la prosa y de la razón. 4. Lima, el Paraíso en la Tierra En la «Aprobación», cuando Bermúdez da cuenta del octavo canto, del cielo estrellado, no queda duda de la íntima relación con la ciudad de Lima: En el Octavo Canto se describe adornada de todas las galas poéticas y geographicas esta gran Corte del Reyno del Perú en su Corografía, Topografía, Delineación, y Planta, ajustada a la luciente Fábrica del Firmamento, o Cielo Octavo, cuya Esfera resplandece poblada de todas las estrellas fixas […] componiendo una celeste luminosa Ciudad, habitada de innumerables Soles […] [La musa Urania] contempla sus movimientos [de las Estrellas] y observa los influxos de sus astros. En la astrología ptolemaica el octavo cielo se encuentra regido por las estrellas fijas, las que «aparecen siempre en la misma posición unas respecto a otras y que son visibles a la salida o a la puesta del sol, dependiendo de su posición respecto de ellas» (Warburg, 2005, p. 26). En la Divina Comedia, de Dante Alighieri, el Paraíso está dividido en función de la máquina de los cielos ptolemaica. El poeta y Beatriz ascienden por los siete planetas hasta llegar al cielo de las estrellas fijas u octava esfera, el cielo de los espíritus triunfantes. Allí Dante queda ciego momentáneamente al contemplar a san Juan Apóstol y, luego de ser interrogado, recupera la vista para toparse con Adán. Ante tal encuentro, Dante no puede con la curiosidad que brota por sus ojos y Adán lo interpela: «Quieres saber en qué año Dios me puso/en el jardín excelso en que tu guía/a tan larga escalera te dispuso, / y cuánto deleitó a la vista mía/ y qué causa enojó tanto a mi dueño/ y el idioma que hablaba, y lo que hacía» (Alighieri, 2004, Canto XXVI, pp. 109-114). Estas inquietudes encontraron respuesta, pero también siguieron latentes y serán respondidas en el cielo estrellado de Lima. 120 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires Desvío Ernst Robert Curtius (2017, pp. 280-286) tipifica los tópicos usuales en la literatura europea medieval en los que encontró un tema retórico-poético que, como un bajo continuo, se escuchó desde el Imperio romano hasta el siglo XVI. El locus amoenus, o paraje ameno, es el escenario bucólico en el que ha de hallarse un paisaje hermoso, un árbol, un prado, una fuente o arroyo, las aves y su canto, las flores y una suave brisa. El paraje ameno en medio de un bosque salvaje indica al menos un movimiento, un viaje de los protagonistas que se adentran en el descubrimiento del lugar de los lugares. La épica filosófica de fines del siglo XII adopta el locus amoenus en las descripciones del Paraíso terrenal. Curtius (2017, p. 283) encuentra que «Juan de Hanville nos lleva a la fabulosa isla de Thyle, donde se reúnen los filósofos de la antigüedad en un paraje de eterna primavera (SP, I, p. 326)». El clima del Paraíso es el de la primavera. El médico Unanue citó al unísono a todos los viajeros de Europa que sostienen que el clima de la ciudad de Lima es el del Paraíso, razón por la que cita, en su escrito de 1806, el canto VIII, en su cantiga III del poema Lima fundada. 5. Retorno adánico Sin intentar realizar un estudio de las influencias de la poética de Dante Alighieri en la obra de Peralta Barnuevo, se puede aventurar que la maquinaria celeste ordenaba los cantos en ambos poemas. Además, las preguntas que Dante le realizó tácitamente a Adán se respondieron también en Lima —o a estas alturas, en el Paraíso limeño—. La primera inquietud que Adán responde en la Divina Comedia es sobre el año en que Dios lo puso en el Paraíso. Es, de alguna manera, preguntar por el acta de fundación de aquel locus amoenus. En el poema Lima fundada puede leerse: IX Después de siglos quince el Sol media Lustros siete al feliz natal divino, Cuando porque habitar no convenía Las fundaciones del primer destino Al Tello, al Díaz el héroe envía Al Don Benito, a que en fiel más fino Pese el sitio; porque en su alabanza Antes que el pueblo, funde su esperanza Una vez más, las notas que incorpora Peralta llegan al auxilio del lector desprevenido para indicar el año de la fundación de Lima, es decir, 1535. El 8 de enero de ese año, Pizarro envía a 121 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez Ruy Díaz, Juan Tello y Alonso Martín de Don Benito para que reconocieran el temperamento, fecundidad, río, leña y puerto, a fin de hacer en él la fundación de su ciudad, trasladando a ella la que había comenzado en Jauja, y después en Sangallan; por no poderse mantener en estas partes sus vecinos, y por la distancia del Mar. El tópico del paraje ameno al que se llega luego de cruzar la selva se encuentra cifrado en la nota de Peralta, así como en estos versos: X Van los de esta del Cielo prometida Feliz fecunda tierra exploradores; De la campaña inculcan estendida Aires, aguas, boscages, frutos y flores: En la orilla, que dejan ya inquirida De ondas y vientos notan los favores Y si su ardor pudiese, allá subiera A indagar las estrellas en la esfera. Se puede seguir respondiendo la inquisitoria adánica sobre la lengua y rastrear en las notas al pie de la obra de Peralta el problema de nombrar, de dar sentido al mundo exultante que rodea a Pizarro, de utilizar el lenguaje para llamar a las cosas que lo circundan, pero ello nos obligaría a tomar nuevos desvíos. El cielo y la poesía limeños estaban en la letra impresa que se leyó en Buenos Aires. 6. Conclusión provisoria La puerta de entrada fue la medicina tropológica en la que el clima y el sujeto guardan una íntima relación; también es motivo de estudio la influencia de los astros en la salud. La América hispánica tuvo en Lima exponentes como José Manuel Dávalos, un rico mulato que egresó de la Universidad de Montpellier en 1787 con su disertación titulada De morbis nonnullis Limae grassantibus ipsorumque therapeia, en la cual, además de limpiar las infamias con las que De Pauw había ensuciado el honor de Lima, sostenía que el clima de esta ciudad es […] salubérrimo, y que no se pueden imputar a él las enfermedades de los limeños. Éstas, según dice, se deben a sus alimentos grasos, viscosos y pesados, al comer mucho y mal, a la costumbre de llenarse de carne de puerco, de papas (que, a lo que sospecha Dávalos, contienen cierto elemento nocivo), de yuca (que, cruda es un veneno) y de aromas y especies picantes. (Gerbi, 1960, p. 265). 122 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires A pesar de la disputa que años después tendría con Hipólito Unanue, ambos estaban de acuerdo en defender la benignidad del clima de Lima tal como el polígrafo Pedro Peralta Barnuevo había escrito años antes en su poema. El clima limeño es destino de varios recorridos que pueden comenzar con la literatura de viaje y sus cronistas, los textos médicos, los panegíricos religiosos y la poesía épica del barroco tardío que aquí se analizó. Se podría sostener que el uso de metáforas, en las que la ciudad de Lima encuentra su paralelo con el Paraíso, es un lugar frecuente y que en manos de los criollos ilustrados sirvió, tal vez provisoriamente, como una reacción identitaria ante la Historia Natural europea enmarcada en el clásico experimento de Montesquieu con la lengua de carnero que tomaba de la Historia Natural del conde de Buffon. Unanue matizó el determinismo climático que cuestionaba a los americanos. Podría aventurarse que se volvió a narrar la conquista del Perú en las notas al pie que Peralta escribió al interpelar las fuentes escriturarias a las que tuvo acceso. Tal vez respondían las mismas preguntas que Dante le hizo a Adán. Podría, sobre la arena, a la espera de las olas, escribirse que el Paraíso como paraje ameno que se encontró en el poema Lima fundada y en Observaciones sobre el clima es una marca fuerte que sobrevivió a la revolución copernicana. La utilización de la astrología y el ordenamiento pagano del cielo estrellado no fue un susurro que se transmitiera entre generaciones, sino que mutó de tal manera que fuera también aceptado por la Iglesia y sus flamígeros inquisidores. Qué portentos guardaba tal concepción del mundo que sirvió para que los criollos limeños narrasen su historia y cuidaran de su salud. Dos libros bajo el mismo cielo tuvieron a la ciudad de Lima como protagonista y ambos fueron leídos más allá de sus fronteras cuando fueron donados para fundar la Biblioteca Pública de Buenos Aires. 123 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022 Matías Maggio-Ramírez Referencias bibliográficas Agamben, G. (2007). La potencia del pensamiento. Adriana Hidalgo. Alighieri, D. (2004). Divina Comedia. [Introducción, traducción y notas de Ángel Crespo]. RBA. Covarrubias, S. de (2006). Tesoro de la lengua castellana o española, Iberoamericana Vervuert. Curtius, E. R. (2017). Literatura europea y Edad Media latina. Fondo de Cultura Económica. Feijoo, B. (s.f.). Teatro crítico universal. http://www.filosofia.org/feijoo.htm Gerbi, A. (1960) La disputa del Nuevo Mundo. Fondo de Cultura Económica. Gutiérrez, J. M. (1957). Escritos coloniales americanos. [Edición, prólogo y notas de Gregorio Weinberg]. Raigal. Lewis, C. S. (1997). La imagen del mundo. Introducción a la literatura medieval y renacentista, Península. Mazzotti, J. A. (2006). La «Aprobación» de Pedro Bermúdez a la Lima fundada: un microcosmos limense. En K. Kohunt, Kart y S. Rose (Eds.), La formación de la cultura virreinal III. El siglo XVIII (pp. 471-486). Iberoamericana Vervuert. Parada, A. E. (2009). Los orígenes de la Biblioteca Pública de Buenos Aires: antecedentes, prácticas, gestión y pensamiento bibliotecario durante la Revolución de Mayo (1810-1826). Instituto de Investigaciones Bibliotecológicas de la Universidad de Buenos Aires (INIBI FFyL-UBA). Peralta Barnuevo, P. (1863). Lima fundada. En M. de Odriozola (Ed.), Colección de documentos literarios del Perú. Establecimiento de tipografía y encuadernación de Aurelio Alfaro. Primeras donaciones de libros a la Biblioteca Nacional de Buenos Aires. (1944). Revista de la Biblioteca Nacional, 30, 493-504. Rico, F. (1997). El sueño del humanismo. De Petrarca a Erasmo. Alianza. Sala Catalá, J. (1994). Ciencia y técnica en la metropolización de América. Doce Calles. Swift, J. (1974). Escritos subversivos. Corregidor. 124 FÉNIX. REVISTA DE LA BIbLIOTECA NACIONAL DEL PErÚ, N°50, 2022 Lima estrellada. Una lectura de las obras de Pedro Peralta Barnuevo e Hipólito Unanue en la fundación de la Biblioteca Pública de Buenos Aires Unanue, H. (1815). Observaciones sobre el clima de Lima. Imprenta de Sacha. Warburg, A. (2005). El renacimiento del paganismo. Aportaciones a la historia cultural del Renacimiento europeo. Alianza. 125 Fénix. Revista de la Biblioteca Nacional del Perú, N°50, 2022
https://openalex.org/W3137825840	https://www.nature.com/articles/s41598-021-85016-9.pdf	English	null	Comparative analysis of machine learning algorithms for computer-assisted reporting based on fully automated cross-lingual RadLex mappings	Scientific reports	2,021	cc-by	15,096	www.nature.com/scientificreports www.nature.com/scientificreports Comparative analysis of machine learning algorithms for computer‑assisted reporting based on fully automated cross‑lingual RadLex mappings OPEN Computer-assisted reporting (CAR) tools were suggested to improve radiology report quality by context-sensitively recommending key imaging biomarkers. However, studies evaluating machine learning (ML) algorithms on cross-lingual ontological (RadLex) mappings for developing embedded CAR algorithms are lacking. Therefore, we compared ML algorithms developed on human expert- annotated features against those developed on fully automated cross-lingual (German to English) RadLex mappings using 206 CT reports of suspected stroke. Target label was whether the Alberta Stroke Programme Early CT Score (ASPECTS) should have been provided (yes/no:154/52). We focused on probabilistic outputs of ML-algorithms including tree-based methods, elastic net, support vector machines (SVMs) and fastText (linear classifier), which were evaluated in the same 5 × fivefold nested cross-validation framework. This allowed for model stacking and classifier rankings. Performance was evaluated using calibration metrics (AUC, brier score, log loss) and -plots. Contextual ML-based assistance recommending ASPECTS was feasible. SVMs showed the highest accuracies both on human-extracted- (87%) and RadLex features (findings:82.5%; impressions:85.4%). FastText achieved the highest accuracy (89.3%) and AUC (92%) on impressions. Boosted trees fitted on findings had the best calibration profile. Our approach provides guidance for choosing ML classifiers for CAR tools in fully automated and language-agnostic fashion using bag-of-RadLex terms on limited expert-labelled training data. There are no studies available that evaluate machine learning (ML) algorithms on cross-lingual RadLex map- pings to provide guidance when developing context-sensitive radiological reporting tools. Therefore, the goal of our study was to compare the performance of ML algorithms developed on features extracted by human experts against those developed on fully automated cross-lingual RadLex mappings of German radiological reports to English1, in order to assist radiologists in providing key imaging biomarkers such as The Alberta Stroke Programme Early CT Score (APECTS)2. We show that this fully automated RadLex-based approach is highly accurate even if the ML models were trained on limited and imbalanced expert labelled data sets3–6. Hence, this work provides a valuable blueprint for developing ML-based embedded applications for context-sensitive computer-assisted reporting (CAR) tools7–10. g RadLex is a comprehensive hierarchical lexicon of radiology terms that can be utilized in reporting, decision support and data mining3. RadLex is freely available (v.4.0, http://radlex.org/) from the Radiological Society of North America (RSNA). It provides the foundation for further ontologies and procedural data bases such as the LOINC/RSNA Radiology Playbook11 or Common Data Elements (CDE; RadElement; https://www.radelement 1Department of Neuroradiology, Medical Faculty Mannheim, Heidelberg University, Theodor‑Kutzer‑Ufer 1‑3, 68137 Mannheim, Germany. Comparative analysis of machine learning algorithms for computer‑assisted reporting based on fully automated cross‑lingual RadLex mappings OPEN 2Department of Biomedical Informatics at the Center for Preventive Medicine and Digital Health (CPD‑BW), Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany. 3Empolis Information Management GmbH, Kaiserslautern, Germany. email: maros@uni-heidelberg.de \| https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 www.nature.com/scientificreports/ Figure 1. The 5 × fivefold nested cross validation setup, which was used to evaluate all machine learning (ML) algorithms and to train the second layer model as a meta/ensemble-learner on top of the combined predictions of these base ML classifiers. Human experts had access to both the findings and impression sections as well as the clinical question field of the reports to generate target labels ASPECTS recommended “yes” (n = 154) vs. “no” (n = 52) and to extract clinico-radiologically relevant features (HEAF). The findings and the impressions were each passed through a fully automated cross-lingual (German-English) natural language processing (NLP) pipeline to generating RadLex mappings. The pipeline can be accessed at https://mmatt.shinyapps.io/rasp/. In order to prevent information leakage, the second layer meta/ensemble models (random forests [RF] and boosted trees [XGBoost]) were trained on the combined inner fold test (i.e. sum of nested validation ΣNtest_1.1–1.5) sets. These second layer models were used to derive objective importance rankings of the individual ML classifiers. To ensure direct comparability between the investigated ML-algorithms, the data partitioning was identical (i.e. each model was trained and fitted on the very same subsamples of the data). However, fastText was fitted directly on German report texts (), whereas other ML-algorithms were fitted on both HEAF and NLP-based RadLex mappings. The final performance measure of the classifiers was calculated as the fivefold cross-validated average on the outer folds (see Tables 1, 2 and 3). ▸ org/12. The official translation of RadLex to German by the German Society of Radiology (DRG) was made public n January 2018 and contained over 45,000 concepts. y p ASPECTS was chosen for this study as a key radiological biomarker, as it is widely used in neurological emergencies to assess the extent of early ischemic changes on pretreatment non-contrast CT studies of the brain in patients with acute ischemic stroke of the anterior circulation2. It proved to be a robust and highly significant independent imaging biomarker to select patients for neurointerventional procedures13. Radiological textual metadata is of crucial importance when selecting patient cohorts for clinical trials or extracting their imaging retrospectively to develop applications using artificial intelligence (AI)14–17. Comparative analysis of machine learning algorithms for computer‑assisted reporting based on fully automated cross‑lingual RadLex mappings OPEN Hence, it is in the best interest of radiologists to report key radiological biomarkers like ASPECTS or other scoring systems to optimize down- stream analytics and software development18,19. Nonetheless, these key predictors are frequently missing from radiological reports as their overwhelming majority is still created as conventional narrative “free-text”1,20,21. In this work, we aim to provide blueprints for creating ML-based CAR tools using a domain-specific ontology to help radiologists improve the content of key biomarkers without disrupting their preferred “free-text” reporting workflow. l ML methods have been introduced as powerful computer-aided diagnostic (CAD) tools9,15,22 in medical image analysis and in radiological reporting23,24. Recently, complex deep transformer-based language models (TLM) are becoming the state-of-the-art (SOTA) in natural language processing (NLP)25–29. However, these models need considerable amount of general and domain specific corpora for training, which are scarce for languages other than English, particularly in the medical domain where creating expert-labelled high-quality training data is extremely resource intensive30–33. Despite achieving SOTA on certain classification tasks, TLMs represent black box methods and show susceptibility to subtle perturbances31,32. Additionally, TLMs are seldom compared to baseline information retrieval methods such as shallow ML algorithms or linear classifiers (fastText) developed on bag-of-words (BOW)34–36. Therefore, we performed comprehensive analyses using an ensemble learning frame- work (Fig. 1) that combined well-established ML algorithms as base classifiers including random forests (RF)37, regularized logistic regression (ELNET)38,39, support vector machines (SVM)40 and classification- (CART)41 and boosted trees (XGBoost)42 as well as fastText36 on German computed tomography (CT) reports with suspected stroke and on their cross-lingual English RadLex mappings using NLP43.lti g g pp g g Our goal was to evaluate a flexible open-source pipeline to swiftly develop robust ML classifiers for CAR tools in a language-agnostic fashion by using cross-lingual bag-of-RadLex mappings on limited expert labelled training data. We aimed to demonstrate the feasibility of our approach by automatically developing production- ready ASPECTS classifiers for CT stroke workups (“MyReportCheck”, Suppl. Fig. S1 online) and compare its performance to ML models that were developed on human expert annotations. Results I Accuracy#: the averaged fivefold CV accuracy is calculated, ACC: accuracy, AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees, CART: classification and regression trees; CT: classification tree; cp: complexity parameter used for CART node splitting (for this no optimization (pruning) was performed); ln(2) ~ RF: column sampling (i.e. bootstrap) representing the settings equivalent to running RF in the xgboost library, [R]: R statistical software environment. Performance of machine learning algorithms developed on human expert‑annotated features (HEAF). CART demonstrated a fivefold CV accuracy of 73.3% with the worst 63% AUC, BS (0.37) and LL (0.87) values among the tested ML-classifiers (Table 1).hiii Performance of machine learning algorithms developed on human expert‑annotated features (HEAF). CART demonstrated a fivefold CV accuracy of 73.3% with the worst 63% AUC, BS (0.37) and LL (0.87) values among the tested ML-classifiers (Table 1).hiii gi The default (“vanilla”) RF classifier fitted on the 28 HEAF achieved a fivefold CV accuracy of 81.5% with an AUC of 82% and corresponding BS and LL of 0.27 and 0.44, respectively (Table 1). Drastically reducing the feature space of vRF to only the nine (9/28: 32.1%) or five (5/28; 17.9%) most important predictors, had a compa- rably limited effect on the predictive performance of vRF: its accuracies decreased 12.8% and 7.7%, respectively; AUC decreased by ~ 16%; while BS (~ 37%) and LL (~ 27%) scores increased (Table 1).i y Fine tuning the RF classifier using the BS (tRFBS) and LL (tRFLL) metrics slightly improved the overall accu- racy without relevantly changing the calibration metrics of the vRF algorithm (Table 1). On the outer folds, both tRFBS and tRFLL limited the feature space similarly – to the 14 or 25–28 most important variables. Interestingly, ME-optimized RF (tRFME) achieved a slightly worse overall performance profile. Results I Report section Method ML Classifier HEAF feature space Rank Software Optimized metric Tested hyperparameter space Selected number of features or hyperparameter settings on outer fold 1.0–5.0 Accuracy# [min–max; %] ME AUC BS LL Human Expert- Annotated Fea- tures (HEAF) CART CT p = 28 (all) rpart [R] ACC rpart.con- trol = default; cp = 0.01 no optimization (no pruning) 28 73.3 [66.7–79.2] 0.27 0.63 0.37 0.87 vRF RF p = 28 (all) 4 randomForest [R] ME ntree = 500, mtry = 5, pvarsel = 28 28 81.5 [73.8–92.7] 0.18 0.82 0.27 0.44 vRF RF p = 28 (all) randomForest[R] ME ntree = 500, mtry = 5, pvarsel = 9 9 71.0 [59.5–82.9] 0.29 0.69 0.37 0.56 vRF RF p = 28 (all) randomForest[R] ME ntree = 500, mtry = 5, pvarsel = 5 5 75.2 [68.3–83.3] 0.25 0.69 0.36 0.54 tRFBS RF p = 28 (all) 2 randomForest[R] BS ntree = [100, 200, 300, … , 900, 1000] 28, 14, 14, 14, 14 83.1 [76.2–90.2] 0.17 0.81 0.27 0.44 tRFME RF p = 28 (all) randomForest[R] ME mtry = [3, 4, 5, 6, 7] 28, 28, 14, 5, 14 79.6 [68.3–90.2] 0.20 0.79 0.29 0.46 tRFLL RF p = 28 (all) 2 randomForest[R] LL pvarsel = [3, 5, 10, 14, 20, 25, 28] 25, 14, 14, 14, 14 83.1 [76.2–90.2] 0.17 0.81 0.27 0.44 ELNET ELNET p = 28 (all) 3 glmnet[R] ME α = [0, 0.1, 0.2, … , 0.8, 0.9, 1] λ = ten- fold CV with default hot-start α = [0.1, 0.8, 0, 1, 0.1] λ = [0.195, 0.0688, 0.208, 0.0301, 0.1632] 82.0 [78.6–85.4] 0.18 0.79 0.27 0.43 SVM-LK SVM p = 28 (all) 1 e1071[R] ME C = [0.001, 0.01, 0.1, 1, 10, 100, 1000] C = [1, 1, 100, 10, 10] 87.4 [82.9–90.2] 0.13 0.79 0.22 0.37 XGBoost BT p = 28 (all) 5 xgboost[R] ME nrouds/ntree = 100, nrouds = 100 80.6 [75.0–85.7] 0.19 0.70 0.30 0.48 max_depth = [3, 5, 6, 8] max_depth = [5, 3, 5, 8, 3] eta = [0.1, 0.3] eta = [0.1, 0.1, 0.1, 0.3, 0.1] gamma = [0, 0.5, 1.0] gamma = [0, 0.5, 1, 0.5, 1] colsam- ple_bytree = [0.1, 0.25, 0.5, 0.693 (ln2) ~ RF, 1] colsample_ bytree = [1, 1, 0.5, 1, 0.5] Table 1. Summary table of performance measures of the investigated ML algorithms developed on human expert-annotated features (HEAF). Results I Inter‑rater reliability of human experts. Providing ASPECTS in the report would have been recom- mended by R1 in 156 (75.7%), by R2 in 154 (74.8%) and by R3 in 155/206 (75.2%) of the cases. The overall agree- ment between the three readers for “ASPECTS recommended” was kappaLight = 0.747 (n = 206, z = 4.6, p = 4.3 × 10–6). The pairwise Cohen’s kappa between R1 and R2 was 0.635 (p < 2 × 10–16), which corresponded to 86.4% agreement. Between R1 and R3 it was 0.62 (p < 2 × 10–16) corresponding to 85.9% agreement. Ratings of two (R2 and R3) experienced readers showed an almost perfect alignment kappa = 0.987 (p < 2 × 10–16) with 99.5% overall agreement. Reliability between automated RadLex mappings and expert‑annotated labels. In this ran- dom subsample, which represents a robust cross-section of the daily praxis, ASPECTS was reported extremely rarely in 4/206 (1.9%). Three of which occurred both in the findings and impressions (3/4, 75%) section and one of which was only reported in the impression (1/4, 25%). The RASP tool correctly annotated all ASPECTS-neg- ative (203/203) and ASPECTS-positive (3/3) finding sections. In the impressions, it misclassified one ASPECTS- positive (1/4, 25%) report as negative (1/206, 0.49%). Scientific Reports \| (2021) 11:5529 \| https://doi.org/10.1038/s41598-021-85016-9 www.nature.com/scientificreports/ www.nature.com/scientificreports/ https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ Table 1. Summary table of performance measures of the investigated ML algorithms developed on human expert-annotated features (HEAF). Accuracy#: the averaged fivefold CV accuracy is calculated, ACC: accuracy, AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees, CART: classification and regression trees; CT: classification tree; cp: complexity parameter used for CART node splitting (for this no optimization (pruning) was performed); ln(2) ~ RF: column sampling (i.e. bootstrap) representing the settings equivalent to running RF in the xgboost library, [R]: R statistical software environment. Results I Notably, on the outer fold 4.0, it limited the feature space to only the five RadLex terms.ii i ELNET showed a similar performance profile to RFs when fitted on the 28 HEAF but it achieved a narrower fivefold CV confidence range of its accuracies (78–86%) while obtaining similar AUC, BS and LL scores (Table 1). The mixing parameter alpha (α) was chosen 3 out of 5 times to fit ridge (0) or ridge-like (0.1, 0.1) models while twice to fit lasso (1) or lasso-like (0.8) models on the outer folds.i i ( ) ( ) On HEAF linear kernel SVMs (SVM-LK) achieved the highest fivefold CV accuracy (87.4%) and lowest BS (0.22) and LL (0.37) scores while obtaining a similar AUC of ~ 80% to other ML classifiers (Table 1). Results I The tuning Scientific Reports \| (2021) 11:5529 \| https://doi.org/10.1038/s41598-021-85016-9 www.nature.com/scientificreports/ Report section Method ML Classifier RadLex feature space Rank Software Optimized metric Tested hyperparameter space Selected number of features or hyperparameter settings on outer fold 1.0–5.0 Accuracy# [min–max; %] ME AUC BS LL Findings CART CT p = 907 5 rpart [R] ACC rpart.con- trol = default; cp = 0.01 p = 907 77.2 [70.8–82.7] 0.23 0.74 0.32 0.66 vRF RF p = 300 (us var.filt.) randomForest [R] ME ntree = 500, mtry = 30, pvarsel = 200 pvarsel = 200 76.2 [71.4–85.4] 0.24 0.78 0.33 0.51 vRF RF p = 907 (all) randomForest[R] ME ntree = 500, mtry = 30, pvarsel = 200 pvarsel = 200 72.8 [67.5–78.6] 0.27 0.78 0.33 0.50 vRF RF p = 907 (all) randomForest[R] ME ntree = 500, mtry = 30, pvarsel = 20 pvarsel = 20 71.4 [62.5–75.6] 0.29 0.74 0.40 0.63 tRFBS RF p = 907 (all) randomForest[R] BS ntree = [200, 400, 600, … , 1400, 1600] pvarsel = [500, 50, 100, 100, 50] 75.2 [71.4–81.0] 0.25 0.76 0.33 0.51 tRFME RF p = 907 (all) randomForest[R] ME mtry = [20, 25, 30, 35, 40] pvarsel = [907, 907, 907, 907, 907] 74.3 [67.5–83.3] 0.26 0.77 0.33 0.50 tRFLL RF p = 907 (all) randomForest[R] LL pvarsel = [10; 20; 50; 100; 200; 500; 907] pvarsel = [50, 50, 50, 50] 75.7 [70.7–85.7] 0.24 0.77 0.33 0.52 ELNET ELNET p = 907 (all) 4 glmnet[R] ME α = [0, 0.1, 0.2, …, 0.8, 0.9, 1] λ = tenfold CV with default hot-start α = [0.2, 0.7, 0.9, 1, 0.1] λ = [0.2685, 0.134, 0.0793, 0.114, 0.397] 79.6 [76.2–82.9] 0.20 0.75 0.29 0.46 SVM-LK SVM p = 907 (all) 3 e1071[R] ME C = [0.001, 0.01, 0.1, 1, 10] C = [0.1, 0.1, 0.1, 0.1, 0.1] 82.5 [78.6–85.4] 0.18 0.80 0.27 0.43 XGBoost BT p = 907 (all) 1 xgboost[R] ME nrouds/ ntree = 100, nrouds = 100 85.4 [80.9–90.2] 0.15 0.78 0.25 0.45 max_depth = [3, 5, 6, 8] max_depth = [5, 8, 5, 8, 3] eta = [0.1, 0.3] eta = [0.1, 0.1, 0.1, 0.3, 0.1] gamma = [0, 0.5, 1.0] gamma = [0, 0.5, 1, 0.5, 1] colsample_ bytree = [0.1, 0.25, 0.5, 0.693 (ln2) ~ RF, 1] colsample_ bytree = [1, 1, 0.5, 1, 0.5] fastText linear direct fit on text 2 Fasttext [Python] ACC & LL default – 83.0 [81.0–85.4] 0.17 0.81 0.29 0.98 Continued https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ parameter of C was selected as 1 on two outer folds suggesting a larger margin for the separating hyperplane while Report section Method ML Classifier RadLex feature space Rank Software Optimized metric Tested hyperparameter space Selected number of features or hyperparameter settings on outer fold 1.0–5.0 Accuracy# [min–max; %] ME AUC BS LL Impressions CART CT p = 675 4 rpart [R] ACC rpart.con- trol = default; cp = 0.01 p = 675 85.0 [79.3–89.5] 0.15 0.75 0.26 0.58 vRF RF p = 300 (us var.filt.) randomForest [R] ME ntree = 500, mtry = 26, pvarsel = 200 pvarsel = 200 83.0 [71.4–88.1] 0.17 0.87 0.25 0.39 vRF RF p = 675 (all) randomForest [R] ME ntree = 500, mtry = 26, pvarsel = 200 pvarsel = 200 82.5 [71.4–88.1] 0.17 0.87 0.25 0.39 vRF RF p = 675 (all) randomForest [R] ME ntree = 500, mtry = 26, pvarsel = 20 pvarsel = 20 78.2 [70–85.4] 0.22 0.81 0.30 0.49 tRFBS RF randomForest [R] BS ntree = [200, 400, 600, …, 1400, 1600] pvarsel = [200, 100, 200, 500, 200] 80.0 [69.0–87.8] 0.20 0.85 0.26 0.41 tRFME RF p = 675 (all) randomForest [R] ME mtry = [21, 26, 31, 36, 41] pvarsel = [200, 675, 200, 675, 500] 83.0 [69.0–90.5] 0.17 0.85 0.25 0.41 tRFLL RF randomForest [R] LL pvarsel = [10; 20; 50; 100; 200; 500; 675] pvarsel = [50, 100, 50, 500, 50] 79.6 [71.4–87.8] 0.20 0.84 0.27 0.42 nodesize = [1; 2 (1%); 10 (5%)] ELNET ELNET p = 675 (all) 3 Glmnet [R] ME α = [0, 0.1, 0.2, …, 0.8, 0.9, 1] α = [0.9, 0.4, 1, 0, 0.9] 85.0 [82.9–88.1] 0.15 0.85 0.22 0.37 λ = tenfold CV with default hot-start λ = [0.056–2.01] SVM-LK SVM p = 675 (all) 2 e1071 [R] ME C = [0.001, 0.01, 0.1, 1, 10] C = [0.1, 0.1, 0.01, 0.1, 0.01] 85.4 [80.0–90.2] 0.15 0.86 0.21 0.36 XGBoost BT p = 675 (all) 5 xgboost [R] ME nrouds/ ntree = 100, nrouds = 100 83.0 [71.4–90.2] 0.17 0.83 0.26 0.44 max_depth = [3, 5, 6, 8] max_depth = [5, 3, 6, 5, 6] eta = [0.1, 0.3] eta = [0.3, 0.3, 0.1, 0.1, 0.3] gamma = [0, 0.5, 1.0] gamma = [0, 0, 1, 0.5, 0.5] colsample_ bytree = [0.1, 0.25, 0.5, 0.693 (ln2) ~ RF, 1.0] colsample_ bytree = [0.1, 0.25, 0.1, 1, 0.1] fastText linear direct fit on text 1 Fasttext [Py] ACC & LL default – 89.3 [832 .3–97.6] 0.11 0.92 0.18 0.55 Table 2. Results I Summary table of performance measures of the investigated ML algorithms on the NLP-annotated bag-of-RadLex features of the findings and impressions sections. Accuracy#: the averaged fivefold CV accuracy is calculated, ACC: accuracy, AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees, CART: classification and regression trees; CT: classification tree; cp: complexity parameter used for CART node splitting (for this no optimization (pruning) was performed); ln(2) ~ RF: column sampling (i.e. bootstrap) representing the settings equivalent to running RF in the xgboost library, [R]: R statistical software environment; [Py] Python v3.7 programming language. Table 2. Summary table of performance measures of the investigated ML algorithms on the NLP-annotated bag-of-RadLex features of the findings and impressions sections. Accuracy#: the averaged fivefold CV accuracy is calculated, ACC: accuracy, AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees, CART: classification and regression trees; CT: classification tree; cp: complexity parameter used for CART node splitting (for this no optimization (pruning) was performed); ln(2) ~ RF: column sampling (i.e. bootstrap) representing the settings equivalent to running RF in the xgboost library, [R]: R statistical software environment; [Py] Python v3.7 programming language. Table 2. Summary table of performance measures of the investigated ML algorithms on the NLP-annotated bag-of-RadLex features of the findings and impressions sections. Results I The calibration profiles of the best performing machine learning classifiers (a–d) fitted on the RadLex mappings and of the random forests meta/ensemble learner (e,f) fitted on the predicted probabilities of the ML-algorithms as features on all outer folds combined (N = 206). Probability estimates for each report by each ML classifier were recorded i.e. how likely it is that the predicted target label is “ASPECTS: yes”. The reliability of these predictions can be assessed visually on calibration plots. Calibration curves are created by grouping reports into discrete bins based on their assigned probability estimates by the ML-model. Thus, the probability space [0–1] gets discretized into bins (i.e. 0–0.1, 0.1–0.2, …, 0.8–0.9, 0.9–1.0; grey grid). The points represent the mean predicted probability (x-axis) and the observed fraction (y-axis) of true (“yes”) labels for the subset of reports falling in that respective range. For ideally calibrated models, the mean predicted probability and observed fraction should be identical within each bin, hence the calibration curve would lie on the diagonal (grey line). Rug plots (blue lines, findings; red lines, impressions) indicate the axis-values of the aforementioned aggregated bin measures (thick lines) and probability estimates of single reports (thin lines). ELNET (a) was more suitable for the impressions (red) particularly in the 0.50–0.75 range, corresponding to its top 3 ranked accuracy. Linear kernel SVMs (b) showed well-calibrated estimates for the 0.50–1.0 probability domain for both the findings (blue) and impressions (red). XGBoost (c) presented an almost ideal calibration curve on the findings (blue) while being the most accurate ML classifier (Table 2). FastText (d) achieved the highest overall accuracy when trained on the impressions (red) with partly well-calibrated estimates (0.75–1) but it was poorly calibrated on the findings (blue). The RF meta/ensemble learner (e) showed a reasonably well-calibrated profile when trained on probability outputs of all ML-algorithms (16 × ML models both findings and impressions; see Table 3). The histogram inset displays the bimodal distribution of its probability estimates. It showed (f) similar calibration profiles when trained either only on 8–8 ML model estimates of the findings (blue) or the impressions (red), respectively. Figure 2. The calibration profiles of the best performing machine learning classifiers (a–d) fitted on the RadLex mappings and of the random forests meta/ensemble learner (e,f) fitted on the predicted probabilities of the ML-algorithms as features on all outer folds combined (N = 206). Results I Probability estimates for each report by each ML classifier were recorded i.e. how likely it is that the predicted target label is “ASPECTS: yes”. The reliability of these predictions can be assessed visually on calibration plots. Calibration curves are created by grouping reports into discrete bins based on their assigned probability estimates by the ML-model. Thus, the probability space [0–1] gets discretized into bins (i.e. 0–0.1, 0.1–0.2, …, 0.8–0.9, 0.9–1.0; grey grid). The points represent the mean predicted probability (x-axis) and the observed fraction (y-axis) of true (“yes”) labels for the subset of reports falling in that respective range. For ideally calibrated models, the mean predicted probability and observed fraction should be identical within each bin, hence the calibration curve would lie on the diagonal (grey line). Rug plots (blue lines, findings; red lines, impressions) indicate the axis-values of the aforementioned aggregated bin measures (thick lines) and probability estimates of single reports (thin lines). ELNET (a) was more suitable for the impressions (red) particularly in the 0.50–0.75 range, corresponding to its top 3 ranked accuracy. Linear kernel SVMs (b) showed well-calibrated estimates for the 0.50–1.0 probability domain for both the findings (blue) and impressions (red). XGBoost (c) presented an almost ideal calibration curve on the findings (blue) while being the most accurate ML classifier (Table 2). FastText (d) achieved the highest overall accuracy when trained on the impressions (red) with partly well-calibrated estimates (0.75–1) but it was poorly calibrated on the findings (blue). The RF meta/ensemble learner (e) showed a reasonably well-calibrated profile when trained on probability outputs of all ML-algorithms (16 × ML models both findings and impressions; see Table 3). The histogram inset displays the bimodal distribution of its probability estimates. It showed (f) similar calibration profiles when trained either only on 8–8 ML model estimates of the findings (blue) or the impressions (red), respectively. with worse LL metrics. On the impressions, however, CART was tied for the 3rd best accuracy (85.0%) but still it showed low AUC (0.75) and high LL (0.58) values.i with worse LL metrics. On the impressions, however, CART was tied for the 3rd best accuracy (85.0%) but still it howed low AUC (0.75) and high LL (0.58) values.i g As for RF, applying unsupervised variance filtering to select the top 33% most variable RadLex mappings of the findings sections, improved the fivefold CV accuracy of vRF by ~ 4.7%. Results I Accuracy#: the averaged fivefold CV accuracy is calculated, ACC: accuracy, AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees, CART: classification and regression trees; CT: classification tree; cp: complexity parameter used for CART node splitting (for this no optimization (pruning) was performed); ln(2) ~ RF: column sampling (i.e. bootstrap) representing the settings equivalent to running RF in the xgboost library, [R]: R statistical software environment; [Py] Python v3.7 programming language. parameter of C was selected as 1 on two outer folds suggesting a larger margin for the separating hyperplane while larger values of 10 or 100 were selected on the remaining three outer folds, suggesting a smaller-margin classifier. Boosted decision trees were similarly accurate (80.6%) like tuned RF and ELNET. Despite the detailed tun- ing grid, XGBoost had overall somewhat worse performance profile than the other investigated ML algorithms, particularly AUC was lower at 70% for which we do not have a clear explanation. parameter of C was selected as 1 on two outer folds suggesting a larger margin for the separating hyperplane while larger values of 10 or 100 were selected on the remaining three outer folds, suggesting a smaller-margin classifier. Boosted decision trees were similarly accurate (80.6%) like tuned RF and ELNET. Despite the detailed tun- ing grid, XGBoost had overall somewhat worse performance profile than the other investigated ML algorithms, particularly AUC was lower at 70% for which we do not have a clear explanation. Performance of machine learning algorithms developed on fully automated RadLex map‑ pings. Directly applying a single classification tree (CART) without optimizing its tree complexity (i.e. no pruning) showed on the findings similar overall accuracy (77.2%) to vRF with similar AUC and BS (Table 2) but https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ Figure 2. www.nature.com/scientificreports/ SVM-LK had the highest AUC and lowest LL on the findings while on the impressions, it was overall the best- performing base ML-classifier. SVMs were comparably well-calibrated for both the findings and impressions, especially in the 0.5–1.0 probability domain (Fig. 2b).i p y p y g XGBoost performed particularly well on the RadLex mappings of the findings – where the other ML algo- rithms (including fastText) struggled (Table 2). It showed the highest accuracy (pAcc.vs.NIR = 1.4 × 10–4) and low- est BS with corresponding slightly worse AUC and LL metrics (than the runner-up SVM-LK). Nevertheless, it had the best overall visual calibration profile on the reliability diagrams for the whole probability domain (Fig. 2c). Compared to the findings, on the impressions XGBoost tuning implied a stronger subsampling of the features when constructing each tree, thereby strongly limiting the available predictor space. On the impressions, XGBoost performed similar to RF classifiers. Linear models (fastText) fitted directly on German report text. When directly fitting the findings sections of the reports, the fastText algorithm showed a fivefold CV accuracy of 83.0% (95%CI: 77.2–87.9%; pAcc.vs.NIR = 0.0030) with sensitivity of 94.8%, and specificity of 48.1% (PPV 84.4%, NPV: 75.8%), which cor- responded to 84.4% precision and 89.3% F1 score. It achieved comparable AUC (81.1%) and BS (0.29) to other shallow ML-models trained on RadLex mappings but showed markedly worse LL profile (0.98) suggesting “more certain” misclassifications.i i FastText achieved the best results across all investigated ML algorithms fitted on the impressions sections of the reports. It showed a fivefold CV accuracy of 89.3% (95%CI: 84.3–93.2%; pAcc.vs.NIR = 1.35 × 10–7) with a bal- anced accuracy of 82.0%. Its predictive profile was in the 87–97% range (sensitivity: 96.8%; specificity: 67.3%; PPV 89.8%, NPV: 87.5%) with precision of 89.8% and F1 score of 93.1%. Furthermore, it showed the highest AUC (91.7%) with lowest BS (0.18) but yet again somewhat worse LL (0.55) than the RadLex-based ML algorithms. FastText showed poor visual calibration profiles for both the findings and impressions in the lower probability domains (0–0.5), however it was almost ideally calibrated in the 0.75–1.0 domain of the impressions (Fig. 2d). Performance of the second layer meta/ensemble‑learners. The second layer meta/ensemble RF learner, which was trained on predictions of the ML-classifiers of the findings sections, showed similar per- formance metrics (Table 3) as the top single ML-classifiers like SVM-LK, fastText and XGBoost (Table 2). www.nature.com/scientificreports/ Its accuracy was in the 77–88% 95%CI range (pAcc.vs.NIR = 1.8 × 10–4) with 89.6% sensitivity; 65.3% specificity; 88.5% PPV; and 68% NPV which corresponded to a precision of 88.5% and F1 score of 89.6%. SVM-LK was chosen twice as the most important classifier while vRF, ELNET and XGBoost were each selected once on the five other folds (Fig. 3a,d).hi g The fivefold CV accuracy (89.3%) of the ensemble RF (Table 3), when using only the ML-models of the impressions as input features, was identical to the best predictor (fastText). But the 95% confidence interval got narrower and the LL score got considerably reduced (by 38%). This solely impressions-based ensemble achieved the following metrics: sensitivity 92.2%; specificity 80.8%; PPV 93.4%, NPV 77.8% with corresponding preci- sion of 93.4% and F1 score of 92.8%. FastText was chosen as the most important predictor for all outer fold test sets while as top 2nd predictor XGBoost was chosen twice; ELNET, SVM-LK and tRFBS were each selected once, respectively (Table 3; Fig. 3b,e). p y g When the ML-classifier predictions of both the findings and impressions were the combined input for the second layer RF model, its accuracy, BS and LL slightly got worse (5–6%). The confusion matrix derivates were as follows: sensitivity 91.6%; specificity 80.8%; PPV 93.4%, NPV 76.4% with corresponding precision of 93.4% and F1 score of 92.5%. The variable importance rankings were dominated by ML-classifiers developed on the impression sections (Table 3; Fig. 3c,f). The visual calibration profile of the RF ensemble developed on all ML- models (both findings and impressions; p = 16) are presented in (Fig. 2e,f). i g p p p g On this same combined feature space (p = 16), the second layer XGBoost ensemble showed a slightly reduced accuracy and worse calibration profiles than the RF ensemble (Table 3). Its predictive profile was in the 82–92% range (pAcc.vs.NIR = 6 × 10–6; sensitivity: 93.5%; specificity: 69.2%; PPV 90.0%, NPV: 78.3%) with precision of 90% and F1 score of 91.7%. XGBoost selected fastText impressions 3 × and SVM impressions 2 × out of 5 on the outer folds as the most important variable based on the gain metric. Results I In contrast, the same variance filter- ing on the impression sections did not relevantly (0.6%) improve vRF’s accuracy (Table 2). Tuned RF models were slightly more accurate than the default vRF, however, tuning did not improve much upon the remaining calibration metrics.i ELNET was the 3rd best-performing ML algorithm on the RadLex features of the findings sections behind SVMs and XGBoost with similar BS and LL metrics but lower accuracy (pAcc.vs.NIR = 0.061) and AUC (Table 2). On the impression, it achieved the second highest fivefold CV accuracy (85.0%; 95%CI: 79.3–89.5%; pAcc.vs.NIR = 2.8 × 10–4) with corresponding second-best calibration profile (AUC: 86%; BS: 0.22; and LL: 037). On the outer folds of the impressions lasso or lasso-like settings (0.9–1) dominated the tuned α settings. ELNET had a better visual calibration profile on the impressions than on the findings (Fig. 2a).i pi pi g g Linear kernel SVMs (SVM-LK) were the only classifiers that performed in the top 2 on the RadLex feature spaces of both the findings (pAcc.vs.NIR = 5.1 × 10–3) and impressions (pAcc.vs.NIR = 1.4 × 10–4) sections (Table 2). https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ Discussion In this work, we present a resource effective approach to develop production-ready embedded ML models for CAR tools, in order to assist radiologists in providing clinically relevant key biomarkers9,20,44,45. To our knowl- edge, this is the first study that uses fully automated cross-lingual (German to English) RadLex mappings- based machine learning to improve radiological reports by suggesting the key predictor ASPECTS in CT stroke workups. We demonstrated the feasibility of our automated RadLex framework (“MyReportCheck”, Supplemen- tary Fig. S1 online) by comparing it to ML classifiers developed on human expert annotations. Furthermore, our ensemble learning setup provides objective rankings and a generalizable blueprint for choosing ML algorithms when developing classifiers for similar context-sensitive recommendation tasks44,46. p gi Although reporting templates have been developed to promote and standardize the best practice of radio- logical reporting47–49, the majority of radiology reports are still created in free-text format50,51. This limits the use of radiology reports in clinical research and algorithm development45,49,51. To overcome this, NLP pipelines including ML proved to be effective to annotate and to extract recommendations from reports51,52. Nonetheless, studies dealing with ML algorithm development particularly for real-time context-sensitive assistance of radiolo- gists while writing reports are scarce46,53. Therefore, in this work, we focused on comprehensive and objective comparison of ML algorithms to provide technical guidance for developing these algorithms on limited (non- English) training data. For this, we have put an emphasis on the probabilistic evaluation and ranking of ML Scientific Reports \| (2021) 11:5529 \| https://doi.org/10.1038/s41598-021-85016-9 www.nature.com/scientificreports/ Table 3. Summary table of performance measures of the second layer meta/ensemble learners (random forests and boosted trees) combining the predictions of all RadLex-based ML base classifiers from the findings and impression sections. AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM, n.SV: number of support vectors; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees. Discussion Ensemble ML-algorithm Classifiers Number of features (ML-model outputs) Most important ML-classifiers/ outer fold Optimized metric Hyperparameters Selected number of features or hyperparameter settings on outer fold 1.0–5.0 Accuracy# [95%CI] ME AUC BS LL vRF vRF 8 × ML-models (findings) Top 1: ME ntree = 500, mtry = 2, pvarsel = 8 pvarsel = 8 83.5 [77.7–88.3] 0.17 0.83 0.29 0.47 tRFBS, vRF-find 1/5 tRFME, SVM-find 2/5 tRFLL, ELNET-find 1/5 ELNET, XGBoost 1/5 SVM-LK, XGBoost, fast- Text Top 2: XGBoost-find 1/5 tRF-ME-find 2/5 fasstext-find 1/5 ELNET-find 1/5 vRF vRF 8 × ML-models (impressions) Top 1: ME ntree = 500, mtry = 2, pvarsel = 8 pvarsel = 8 89.3 [84.3–93.2] 0.11 0.90 0.19 0.34 tRFBS, fasstext-impr 5/5 tRFME, Top 2: tRFLL, svm-impr 1/5 ELNET, XGBoost-impr 2/5 SVM-LK, XGBoost, fast- Text tRF-BS-impr 1/5 ELNET-impr 1/5 vRF vRF 16 × ML-models Top 1: ME ntree = 500, mtry = 4, pvarsel = 16 pvarsel = 16 88.8 [83.7–92.8] 0.11 0.90 0.20 0.36 tRFBS, (8 × findings & fasstext-impr 5/5 tRFME, 8 × impressions) Top 2: tRFLL, svm-impr 3/5 ELNET, tRF-BS-impr 1/5 SVM-LK, XGBoost, fast- Text ELNET-impr 1/5 XGBoost vRF 16 × ML-models Top 1: ME nrouds/ntree = [5, 10, 25, 50, 75, 100] nrounds = [75, 5, 75, 5, 10] 87.4 [82.0–91.6] 0.13 0.87 0.30 0.46 tRFBS, (findings & impressions) fasstext-impr 3/5 max_depth = [3, 5, 6, 8] max_depth = [3, 6, 5, 3, 5] tRFME, svm-impr 2/5 eta = [0.01, 0.1, 0.3] eta = [0.3, 0.01, 0.1, 0.01, 0.1] gamma = [1, 0.01, 0.1, 0, 0.5] colsample_ bytree = [0.1, 0.5, ln2~RF, 0.1, 0.25] tRFLL, Top 2: gamma = [0, 0.001, 0.01, 0.1, 0.5, 1] ELNET, fasstext-impr 2/5 colsample_ bytree = [0.1, 0.25, 0.5, 0.693 (ln2) ~ RF, 1.0], SVM-LK, XGBoost, fast- Text tRF-BS-impr 2/5 min_child_ weight = 1, svm-impr 1/5 subsample = 1 Table 3. Summary table of performance measures of the second layer meta/ensemble learners (random forests and boosted trees) combining the predictions of all RadLex-based ML base classifiers from the findings and impression sections. Discussion AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM, n.SV: number of support vectors; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees. Table 3. Summary table of performance measures of the second layer meta/ensemble learners (random forests and boosted trees) combining the predictions of all RadLex-based ML base classifiers from the findings and impression sections. AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM, n.SV: number of support vectors; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees. Table 3. Summary table of performance measures of the second layer meta/ensemble learners (random forests and boosted trees) combining the predictions of all RadLex-based ML base classifiers from the findings and impression sections. AUC: multiclass area under the ROC after Hand and Till (that can only be calculated if probabilities are scaled to 1), us var.filt: unsupervised variance filtering using p = 300 most variable RadLex terms -this step was previous of training to prevent information leakage, BS: Brier score, ME: misclassification error, LL: multiclass log loss, vRF and tRF: vanilla- and tuned random forests, ELNET: elastic net penalized multinomial logistic regression, SVM: support vector machines, LK: linear kernel SVM, n.SV: number of support vectors; XGBoost: extreme gradient boosting using trees as base learners, BT: boosted trees. https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ This is less relevant for biomarker CAR recommendation systems but crucial for automated inference ystems for scores such as BI-RADS54 or PI-RADS18. We used a commercially available NLP pipeline that implements a common approach8,51 comprised of cleans- ing, contextualization and concept recognition as well as negation detection trained explicitly for German and English RadLex mappings1,43. This fully automated approach to generate bag-of-RadLex mappings is advan- tageous compared to standard BOW35 approaches, as it already captures domain-specific knowledge includ- ing negation and affirmation3. Mikolov et al. proposed word2vec to create semantic word embeddings, which gained popularity in the field of radiology5,55. However, word2vec struggles to properly handle out-of-vocabulary words56,57. Thus, it needs to be combined with radiology domain-specific mappings. In contrast, our approach directly generates bag-of-RadLex terms for each report. We then combine all binary RadLex term occurrences in our corpus (separately for findings and impressions) to generate the RadLex-DTMs. Therefore, our pipeline is also more robust for new or missing words e.g. if a new report does not contain certain terms (present in the training corpus), these can be easily substituted with 0 or new terms can be added to the DTM and the ML clas- sifier can be swiftly retrained. This commercial NLP-based RadLex-mapping pipeline for creating DTMs is free for research purposes and can be easily utilized through our Shiny application. Similar to previous studies47,51, we included all hierarchical parent and child elements of the tree structure of RadLex concepts as a flattened feature space and let the ML classifiers select subgroups of terms relevant to the classification task automatically during training. For a similar domain-specific semantic-dictionary mapping, as part of their hybrid word embedding model, Banerjee et al. created a custom ontology crawler that identi- fied key terms for pulmonary embolism57. Another approach by Percha et al. included only partial flattening of RadLex. They selected the eight most frequent parent categories that were used to learn word and RadLex term vector representations for automatically expanding ontologies5. We have also found that certain key terms are missing from RadLex and manually extended it. Other approaches to mitigate this problem and to increase interoperability, aim to combine multiple (both radiology-specific and general medical) ontologies or procedural databases such as RadLex, LOINC/RSNA playbook, CDE from the RSNA and Systematized Nomenclature of Medicine Clinical Terms (SNOMED CT) as well as the International Classification of Diseases (v.10) Clinical Modification (ICD-10-CM)56,58–60. www.nature.com/scientificreports/ www.nature.com/scientificreports/ w.nature.com/scientificreports/ https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ Figure 3. Two corresponding pairwise versions of multi-way importance plots of the investigated machine learning algorithms based on the random forests meta/ensemble learner when fitted on the probability estimates of the eight ML models as features (Table 3) based on the findings (a,b), impressions (c,d) and both (e,f) report sections. The axes on subplots (a,c,e) measure the prediction related relevance of a variable. Here, y-axes (Gini_decrease) display the Gini feature importance-based mean decrease in node impurity while the x-axes (Accuracy_decrease) show the more robust mean decrease in accuracy (type = 1) variable importance measure6,62–64. P-values (legend: red, green and blue patches and colored text brackets) were derived from a binomial distribution of the number of nodes split on the variable assuming random draws. On subplots (b,d,f), y-axes (Times_a_root) show the number of trees in which the root is split on that variable (i.e. ML classifier), whereas the x-axes (Mean_minimal_depth) show the mean depth of first split on the variable. Because these two measures are negatively associated, most important variables are located in the upper-left corner. Area of the points is proportional with the total number of nodes (no_of_nodes) in the forest that split on that variable and the points are blue if the variable was used as root (top). When ML classifiers trained only on the findings sections were fed to the RF ensemble (a), XGBoost (p < 0.01) was the only significant predictor while linear kernel SVM showed a weak trend (p < 0.1). Underscoring XGBoost’s importance (b), it was used in the most nodes and as root split. Among the models developed on the impressions (c), fastText (p < 0.01) was the most important predictor followed by SVM-LK (p < 0.01) while brier score-tuned RF (tRF-BS) showed a week trend (p < 0.1). FastText and SVM-LK (d) were the most relevant classifiers based on tree splitting measures. Likewise, when all 16 ML-models were combined (e), fastText (p < 0.01) and SVM-LK (p < 0.01) based on the impressions dominated the importance rankings, however, although less relevant findings-based XGBoost still achieved a weak trend (p < 0.1). Plots were created on the first outer fold test set (Ntest.1.0 = 42). ◂ classifiers. This is less relevant for biomarker CAR recommendation systems but crucial for automated inference systems for scores such as BI-RADS54 or PI-RADS18. lassifiers. www.nature.com/scientificreports/ findings sections, most ML models including fastText struggled but XGBoost performed best with an almost ideal calibration profile among all models (including those developed on the impressions). As impressions are expert-created condensed extracts of the most relevant information, ML performed substantially better (all > 80%). Accordingly, both RF and XGBoost meta/ensemble learners favored ML models that were developed on the impressions particularly fastText, SVM-LK and BS-tuned RF. These second layer meta/ensemble models achieved precision of 90–93%, recall: 92–94% and F1 score: 91–93%, which was well in line with the performance of information extraction model by Hassanpour et al. on a similarly sized (n = 150) test set of multi-institutional chest CT reports51. p The advantage of RadLex-based ML models compared to fastText is that they contain anatomical concepts and we can directly access negation information providing human interpretable explanation of the model. For fastText, such concepts are not necessarily learnable from limited training data or for more complex decision support scenarios other than ASPECTS. This was also supported by the fact that, despite being a baseline model, single CART performed remarkable well on the impressions implying that recommending ASPECTS is a less complex decision task.h The present study has certain limitations as it was a single-center, retrospective cross-sectional study of limited size. Nonetheless, we tried to create a representative cohort of the general daily praxis by selecting a stratified random sample of ~ 200 reports from ~ 4000 reports from a period of 4 years, which may robustly represent the general daily praxis. Our primary goal was to provide baseline performance metrics for well-established NLP and ML algorithms and linear classifiers with respect to radiology-specific biomarker (ASPECTS) recommendation tasks. Hence, there are natural extensions to our traditional methodology including the switch to well-known neural network architectures at the level of concept recognition to generate RadLex mappings26,67. Recently, DL methods are increasingly used for concept recognition tasks such as long short-term memory (LSTM) and variants of bidirectional recurrent neural networks (BiRNN) coupled with conditional random field (CRF) architectures68,69. DL models can also be used to create task-specific classifiers in an end-to-end manner (e.g., convolutional neural (CNN)24, RNN54 or LSTM networks45,70). However, fastText (with only a single hidden layer) has proven to be on a par with these more complex network architectures on several benchmarks36. www.nature.com/scientificreports/ Although incorporating pre-trained language-specific word representations into fastText was expected to improve its accuracy, we chose not to do so to allow for more direct performance comparisons with bag-of-RadLex-based ML classifiers71. i Utilizing large transformer architectures25,27–29,72 directly on German free-text reports would be a reasonable extension, however, sufficiently large non-English public radiology domain-specific corpora for transfer learning are lacking and the interpretability of TLMs is challenging31. Whether TLMs “truly learn” underlying concepts as a model of language or just extract spurious statistical correlations is a topic of active research32,33. Thus, our CT stroke corpus can facilitate benchmarking of such models for the German radiological domain31,67,72.f g g For recommending ASPECTS we used pyes > 0.5 probability threshold. Optimizing this cutoff could further improve the performance metrics of the ML classifiers – for example by maximizing the Youden index73. i To counteract class imbalance, we also explored upsampling, downsampling, random over-sampling and synthetic minority over-sampling techniques (SMOTE)74, however, they did not improve the accuracy of ML classifiers on our data set (data not shown). i Regardless of these limitations, compared to text-based DL methods, our approach has some major advan- tages: i) building ML classifiers on top of cross-lingual RadLex mappings incorporates domain-specific knowledge thereby only requiring a limited amount of expert labeled data – for which simple class labels may be sufficient; ii) this approach can be easily adopted to any other language where RadLex was translated by the local radiologi- cal society; iii) an ultimate benefit of our methodology is that it allows for the instant interoperability between languages especially the direct transportability of any ML model created for biomarker recommendation or infer- ence from one language to another. Furthermore, the investigated ML algorithms has been proven to be effective for high-dimensional multiclass classification problems in various scientific domains6, therefore, are expected to generalize well for other (more complex) radiological key biomarkers with multiple outputs (e.g., BI-RADS54, PI-RADS18). However, developing classifiers for biomarkers that describe more complicated pathophysiological processes or entities (than ASPECTS) will possibly require lager data sets. p p y q g In conclusion, we showed that expert-based key information extraction and fully-automated RadLex map- ping-based machine learning is comparable and requires only a limited amount of expert-labeled training data – even for highly imbalanced classification tasks. www.nature.com/scientificreports/ i ( ) All investigated ML algorithms were “CPU only” thereby imposing minimal hardware requirements and being quick both at train and test time36. These ML models have proven to be effective on both text classification8,34,36 and other high-dimensional medical problems including high-throughput genomic microarray data6,61. Addition- ally, we implemented a nested CV learning framework in order to objectively assess the importance of each ML base classifier and report section (i.e. findings and impressions) based on their probability estimates of recom- mending ASPECTS6. Zinov et al. also used a probabilistic ensemble learning setup to match lung nodule imaging features to text53. It is of note that there is multicollinearity both on the level of RadLex mappings when training ML base classifiers and when combining the probability estimates of these ML classifiers on the second layer meta/ensemble-learner level. Default settings of RF (both in Python and R) are less robust for these scenarios due to the dilution of true features6,62–64. To counter act dilution, we used the permutation-based importance (type = 1) without scaling for all RF models, which were suggested as the most robust settings in6,63,64. In contrast, boosted trees by design are less susceptible to correlation of features42,65. The performance of the investigated ML algorithms is differently sensitive to the number of features6,61. Based on results by limiting the feature space with unsupervised variance filtering, we suggest using all annotated RadLex features as input and treating the num- ber of features (p) as a tuning parameter during ML-algorithm training to achieve the best possible accuracies. ML models developed on HEAF were similarly accurate (87%) to those developed on fully automated p g p g g g p ML models developed on HEAF were similarly accurate (87%) to those developed on fully automated cross-lingual RadLex mappings (~ 85%), although the latter models had substantially better calibration profiles (especially AUC and BS). This corresponded to results by Tan et al. on lumbar spine imaging when compar- ing rule-based methods to ML models66. On the more heterogeneous and larger RadLex feature space of the https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ www.nature.com/scientificreports/ We performed detailed comparative analyses of well-established ML algorithms and identified those, which are best suited for automated rule learning on bag-of-RadLex concepts (SVM, XGBoost and RF) and directly on German radiology report texts (fastText) through utilizing a nested CV learning framework. This work provides a generalizable probabilistic framework for developing embedded ML algorithms for CAR tools to context-sensitively suggest, not just ASPECTS but any required key biomarker information. Thereby improving report quality and facilitating cohort identification for downstream analyses. www.nature.com/scientificreports/ total of 4022 reports fulfilled the above criteria. After data cleaning, which excluded cases with missing request- ing department, 3997 reports remained. Next, we generated a stratified random subsample (n = 207, ~ 5.2%) based on age (binned into blocks of 10 years), sex (M\|F), year (in which the imaging procedure was performed) and requesting department. During downstream analyses one report was removed because it contained only a reference to another procedure, leaving n = 206 for later analyses (Fig. 1). The extracted reports were all conven- tional free-texts and were signed off by senior radiologists with at least 4 years of experience in neuroradiology. Information extraction by human experts. Three independent readers (R1, experience 3yrs; R2, 7yrs; R3, 10yrs) assessed the clinical questions, referring departments, findings and impressions of the reports. For each report, all readers independently evaluated whether ASPECTS was provided in the report or should have been provided in the report text (necessary: 154, 74.7%; not meaningful: 52, 25.3%]). Further, the two senior experts (R2 and R3) manually extracted clinico-radiologically relevant key features in the context of whether reporting ASPECTS is sensible based on the presence (yes \| no) of ischemia (separately for new infarct demarca- tion and/or chronic post-ischemic defects); bleeding (separately for each of the following entities: intracerebral hemorrhage (ICH), epi- (EDH), subdural hematoma (SDH), subarachnoid hemorrhage (SAH)); tumor; proce- dures including CT-angiography (CTA) or CT-perfusion (CTP); whether cerebral aneurysms or arteriovenous malformations (AVM) were detected; previous neurosurgical (clipping, tumor resection) or neurointerven- tional procedures (coiling); and previous imaging (within the last 1–3 days)75,76. These human expert-annotated features (HEAF) were extracted concurrently from both the finding and impression sections and selected in accordance with national and international guidelines for diagnosing acute cerebrovascular diseases75,76. HEAFs were used as input for ML algorithm development (Table 1). The feature matrix is available as supplementary data (heaf.csv) or GitHub download (https://github.com/mematt/ml4RadLexCAD/data). RadLex mapping pipeline. Both the findings and impression sections of each German report (n = 206) were mapped to English RadLex terms using a proprietary NLP tool, the Healthcare Analytics Services (HAS) by Empolis Information Management GmbH (Kaiserslautern, Germany; https://www.empolis.com/en/). As previously described1,43, HAS implements a common NLP pipeline consisting of cleansing (e.g., replacement of abbreviations), contextualization (e.g. into segments "clinical information", "findings", and "conclusion"), con- cept recognition using RadLex, and negation detection ("affirmed", "negated", and "speculated")77. HAS was pre-trained on ~ 45 k German radiological reports1,43. www.nature.com/scientificreports/ For concept recognition, a full text index and morpho- syntactic operations such as tokenization, lemmatization, part of speech tagging, decompounding, noun phrase extraction and sentence detection were used. The full text index is an own implementation with features such as word/phrase search, spell check and ranking via similarity measures such as Levenshtein distance78 and BM2579. The index is populated with synonyms for all RadLex entities (both from the lexicon and by manual extensions), the morpho-syntactic operations are based on Rosette Base Linguistics (RBL) from Basis Technology (Cam- bridge, MA, USA; https://www.basistech.com/text-analytics/rosette/). For accuracy, RBL uses machine learning techniques such as perceptrons, support vector machines, and word embeddings. For negation detection, the NegEx algorithm was implemented in UIMA RUTA77,80. No further pre-processing steps of the text were done. g g p p p g p Our RadLex annotation and scoring pipeline (RASP), which utilizes the aforementioned HAS API, is freely available as a Shiny application at https://mmatt.shinyapps.io/rasp/35. We used RASP to generate the document (i.e. report RadLex) term matrix (DTM) of the complete data set over all reports (n = 206) both for the findings and impression sections, respectively. In the DTM, each report is represented as a vector (i.e. bag-of-)RadLex terms that occurred in the corpus34,35. All hierarchical parent and child categories of the identified RadLex terms were included as features and encoded in a binary fashion (0\|1), whether the term was present or not. Other kinds of relationships such as “May_Cause” were disregarded. Further, each RadLex term (i.e. feature) was annotated with three levels of confirmation or confidence “affirmed”, “speculated”, “negated”, which was included in the feature name. Feature names were generated by combining the RadLex ID, preferred name of the term and the assigned confirmation level. This DTM provided the basis for fully automated RadLex-based ML algorithm development (Table 2). The report-RadLex term-matrices (i.e. DTMs) both for the findings and impression sections are available for direct download from our GitHub repository (https://github.com/memat t/ml4RadLexCAD/data) or as supplementary data (radlex-dtm-findings.csv and radlex-dtm-impressions.csv).h pp yi g p The performances of ML algorithms developed on these automated NLP-RadLex mappings were then com- pared to those ML algorithms that were developed on the features extracted by human experts (HEAF). It is of note, however, that in its current iteration (v4.0) RadLex does not contain certain key terms or concepts, one of which is ASPECTS. Methods St d h Study cohort. The study was approved by the local ethics committee (Medical Ethics Commission II, Medi- cal Faculty Mannheim, Heidelberg University, approval nr.: 2017-825R-MA). All methods were carried out in accordance with institutional guidelines and regulations. Written informed consents were waived by the ethics committee due to the retrospective nature of the analyses. In this single-center retrospective cohort study, con- secutive (German) radiological reports of cranial CTs with suspected ischemic stroke or hemorrhage between 01/2015–12/2019 were retrieved from local RIS (Syngo, Siemens, Healthineers, Erlangen, Germany) that con- tained the following key words in the clinical < request reason > , < request comment > or < request technical note > fields: “stroke”, “time window for thrombolysis”, “wake up”, “ischemia” and their (mis)spelling variations. A https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ www.nature.com/scientificreports/ Although there is a CDE for ASPECTS classification (https://www.radelement.org/eleme nt/RDE173)12. Hence, extended IDs had to be created for such terms in the NLP annotation service, which are denoted as RadLex ID Extended (RIDE), for example ASPECTS = RIDE172 in the DTMs. Classifiers and feature importance. We performed extensive comparative analyses of well-established ML algorithms (base classifiers) to automatically learn rules required for ASPECTS reporting including single classification (and regression) trees (CART)41, random forests (RF)37, boosted decision trees (XGBoost)42, elas- tic net-penalized binomial regression (ELNET)38,39 and support vector machines (SVM)40. Single CART was used to represent the baseline ML algorithm. A CART has the advantage that human readers can more easily interpret it, however its estimates are much less robust than ensembles of trees like RF41,65,81,82. It is of note that RadLex mappings are inherently correlated features due to RadLex’s hierarchical design. This makes RF suscep- tible to miss the truly relevant terms and dilute the selected features6,62–64. Therefore, we used the most robust metric of permutation-based variable importance (type = 1) without scaling (scale = F) for all RF models6,62–65. Permutation-based variable importance quantifies the importance of a feature by defining a baseline accuracy https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ (for classification tasks) when the initially trained RF model is fitted on the out-of-bag (OOB) samples62,63. Next, all values (observations) of a variable of interest (Xi) are permuted in the OOB samples thereby breaking down any associations between Xi and the outcome. Then, the initial RF model (i.e. each individual tree in the forest) is refitted on this permuted OOB sample and the prediction accuracy is recalculated. The importance of a variable is the difference between the baseline and the drop in overall accuracy after permuting the values of Xi. Notably, the RF classifier is not retrained after permutation, but the already trained baseline model is used to predict on the perturbed OOB sample. Consequently, calculating permutation-based importance metrics for several predictor variables is computationally more expensive than generating the mean decrease in impurity (Gini index) but also proved to be more robust64,83,84. It has also been shown that the raw (unscaled) permutation- based importance measures have better statistical properties83, although they are still potentially biased towards collinear features84. Therefore, we also compared RF to boosted trees, which are by design less susceptible to cor- related features42,65. www.nature.com/scientificreports/ The average fivefold CV model performances on the outer fold test sets are provided in Tables 1, 2 & 3.h p p We chose this nested CV setup to be able to use an independent second layer model. The rationale for this was to investigate whether using the probability outputs of the base ML classifiers as input features for a second layer ensemble model, it could improve the overall performance of suggesting ASPECTS; and to use this “meta/ ensemble” learner to derive importance rankings of the investigated ML algorithms. Hence, we could objectively rank the ML algorithms in addition to comparing their performance metrics. Because these probability esti- mates represented highly correlated features, we chose RF and XGBoost as meta learners (as described above). RF and XGBoost were trained on the combined probability predictions (i.e. “ensemble”) of the base ML models (i.e. CART, RF, XGBoost, ELNET, SVM and fastText) on the respective nested/innerfold test sets (Fig. 1). Then, this tuned model was evaluated on the corresponding outer fold test set preventing any information leakage6. For RF ensemble, we have used mean decrease in accuracy without scaling that has been suggested as the most robust setting when fitting correlated features6,62–64. Importance ranking of boosted decision trees were gener- ated by the gain metric42. Multi-way variable importance plots describing the RF meta learner (Fig. 3) were created using default settings of the “plot_multi_way_importance” function in the randomForestExplainer R package (v0.10.0.)86. Heretofore, we refer to second layer RF and XGBoost algorithms as meta/ensemble learn- ers or models. Text classification directly on German report texts using fastText. We used the open-source, lightweight fastText library (v0.9.1; https://fasttext.cc/) to learn linear text classifiers for ASPECTS recommen- dations on our data set36. The German report texts (both findings and impression sections) were preprocessed by excluding the following special characters “([-.!?,’/()])”. It is of note that fastText was only trained “on-the-fly” in each resampling loop on the corresponding subset of ~ 130–165 reports and we did not utilize any pre-trained word vector model for German71. This approach ensured a more direct comparability with the ML-classifiers developed on bag-of-RadLex mappings. However, pre-trained word vector models for 157 languages, which were pre-trained on Common Crawl and Wikipedia by the fastText package authors are available for direct download (https://fasttext.cc/docs/en/crawl-vectors.html)71. We used the Python (v3.7) interface to fastText (https://github.com/facebookresearch/fastText/tree/master/python) on an Ubuntu 19.10 machine. www.nature.com/scientificreports/ Importance ranking of boosted trees models (both at the annotated feature and meta-learner levels) were derived using the gain metric. Machine learning setup. Each ML algorithm was fitted to the i) human expert-annotated features (HEAF; Table 1) and to the ii) RadLex mapped DTMs both for the findings and impressions separately (Table 2).f Machine learning setup. Each ML algorithm was fitted to the i) human expert-annotated features (HEAF; Table 1) and to the ii) RadLex mapped DTMs both for the findings and impressions separately (Table 2).f Because the effort of manually annotating the data set is large, especially if multiple experts annotate the same reports, we built upon our previously open-sourced protocol of a fivefold nested cross-validation (CV) resampling scheme to have an objective and robust metric when comparing the performance of the investigated methods (Fig. 1). Nested CV schemes allow for the proper training of secondary (e.g. calibrator or ensemble) models, without allowing for information leakage (Fig. 1). To counter act the class imbalance (yes:no = 3:1) dur- ing CV-fold assignment (nfolds.RData), we performed stratified sampling. Also, RFs were downsampled to the minority class during training62,85.i In brief, the data set (n = 206) was divided into stratified subsamples (outer fold training [nouter.train = ~ 164–166] – test set pairs [nouter.test = 40–42]) using fivefold cross-validation (Fig. 1; dashed blue and red boxes). Then, only the outer fold training sets were, yet again, subsampled using fivefold CV, in order to create the nested/inner fold (training [ninner.train = 130–134] – test set pairs [ninner.test = 32–34]; Fig. 1, nested CV). This was performed for both the findings and impressions sections using identical fold structures (Fig. 1).i i Hyperparameter tuning (i.e. training) of the investigated ML algorithms (base classifier) was performed within an extra-nested CV loop on the outer- or inner fold training sets. All models were fitted to the same data structure. Also, random seeds were fixed across all ML algorithms, in order to ensure direct comparability of their performance measures. ML algorithm training was optimized using either accuracy, brier score or log loss, which is indicated along the tuning parameter settings in Tables 2 & 3. For all ML algorithms probability outputs were also recorded and used to measure AUC and to create calibration plots. www.nature.com/scientificreports/ ssess inter-rater agreement whether ASPECTS is recommended in a pairwise fashion for each of the two read- rs. To assess the overall agreement among the three readers, Fleiss’ and Light’s kappa was used. assess inter-rater agreement whether ASPECTS is recommended in a pairwise fashion for each of the two read- ers. To assess the overall agreement among the three readers, Fleiss’ and Light’s kappa was used. Performance was evaluated using calibration metrics focusing on the probabilistic output of the ML base classifiers including the area under the ROC curve (AUC), brier score (BS) and log loss (LL) measures; and derivatives of the confusion matrix: sensitivity, specificity, positive- (PPV) and negative predictive value (NPV) as well as precision, recall and F1 scores. P-values (pAcc.vs.NIR) were provided to quantify the level of accuracy achieved by a ML classifier compared to the no-information rate (NIR) i.e. always predicting only the majority class (154/206, 74.8%). P-values < 0.05 were considered significant. Calibration plots. Calibration plots (or reliability diagrams) are useful graphical tools to visually assess the quality of the probability output of a classifier87,88. Custom functions are available on GitHub (https://githu b.com/mematt/ml4RadLexCAD/tree/master/calibrationplots) to generate calibration plots presented in Fig. 2. Briefly, for real-life problems the true conditional probabilities of target classes are often unknown, therefore the prediction space needs to be discretized into bins88,89. A common approach is to use ten bins (e.g., probability ranges: 0–0.1, 0.1–0.2, …, 0.9–1.0) and assign cases to the corresponding bin where their predicted probabilities by the respective ML classifier fall. Consequently, in each bin there is a distinct subset of the study cohort. For each bin the fraction of true positive cases in that subset (y-axis) is plotted against the mean of the predicted probabilities of the subset by the classifier (x-axis). Hence, the probability output of an ideally calibrated ML classifier would lie on the diagonal line87,89. For instance, if (hypothetically) ELNET estimated the predicted probability of “ASPECTS: yes” between 0.9–1.0 with mean ~ 0.9 for 10 of the reports based on RadLex mappings of their findings and impressions sections, respectively (Fig. 2a, x-axis) and if ELNET was well-calibrated, then the number of reports in which ASPECTS should be truly provided among these 10 reports, would ideally be 9. Hence, the observed fraction of such reports in the cohort (Fig. 2a, y-axis) would be (9/10 = 0.9) identical to the mean prediction6,90. www.nature.com/scientificreports/ The point coordinates representing the mean predicted probability by ELNET (Fig. 2a) and observed fraction in the cohort for this probability bin (0.9–1.0) were, indeed, both very close (red, impressions; blue, findings) and lied almost on the diagonal line87,88. Thus, ELNET was well-calibrated for this bin, but it was poorly calibrated (“unsure”) for the 0–0.25 or 0.5–0.75 ranges as the distance from the diagonal line was larger. Predictions based on the findings or impression varied substantially even with the same ML model (Fig. 2a–f). Data availability y Both the human expert annotated features (heaf.csv) and the fully automated NLP-based RadLex mappings (term-report-matrices) are provided in our GitHub repository (https://github.com/mematt/ml4RadLexCAD/). The RadLex annotation and scoring pipeline (RASP) is freely available for research purposes as Shiny application at www.mmatt.shinyapps.io/rasp . All tuned ML-model objects including the fold IDs for the 5 × fivefold strati- fied nested CV scheme (nfolds.RData) are provided on GitHub. Additionally, we provide R code for ML-model training and for generating calibration plots presented in Fig. 3. Received: 23 September 2020; Accepted: 23 February 2021 www.nature.com/scientificreports/ FastText models were fitted both on the findings and impression sections respectively, using the same 5 × fivefold nested- CV scheme as for the other ML algorithms with similar extra-nested CV loop for training on the outer- or inner fold training sets. Class label predictions and probability outputs were recorded and evaluated in the same man- ner as the investigated ML algorithms developed on HEAF and RadLex mappings. Statistical analyses. All statistical analyses were performed using the R language and environment for statistical programming (R v3.6.2, R Core Team 2019, Vienna Austria). The Cohen’s kappa statistic was used to https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ References 1. Jungmann, F. et al. Towards data-driven medical imaging using natural language processing in patients with suspected urolithiasis Int. J. 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The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; and in the decision to publish the results. The other authors declare no conflicts of interest. https://doi.org/10.1038/s41598-021-85016-9 Scientific Reports \| (2021) 11:5529 \| www.nature.com/scientificreports/ Additional information Supplementary Information The online version contains supplementary material available at https://doi. org/10.1038/s41598-021-85016-9. © The Author(s) 2021 Additional information T Additional information Supplementary Information The online version contains supplementary material available at https://doi. org/10.1038/s41598-021-85016-9. Correspondence and requests for materials should be addressed to M.E.M. Reprints and permissions information is available at www.nature.com/reprints. 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https://openalex.org/W4381195207	https://drpress.org/ojs/index.php/ijeh/article/download/8670/8471	English	null	Research on the Training Mode of "Dual-track Parallel and Fixed Post Training"	International journal of education and humanities	2,023	cc-by	5,840	1.1. Basic connotation of "dual-track parallel, fixed post training" "Collaborative theory" is "synergy theory", which was put forward by Hakken, professor of Stuttgart University and famous physicist in 1971, and was systematically discussed in 1976. It is an important branch theory of systems science. The self-organization principle of synergy theory explains to us that any system that lacks material, energy and information communication with the external environment will be in an isolated or closed state. In this closed state, no matter what the initial state of the system is, eventually any orderly structure inside it will eventually be destroyed, presenting a scene of "dead silence". Therefore, the system can only communicate with the outside world through continuous material, information and energy, to maintain its life and make the system develop in the direction of order. Dual-track that normal colleges and rural primary schools support and cooperate each other for common development, and win-win cooperation promotes the development and development of primary school teachers. Universities to the actual education of rural primary school as reference, timely modification to the cultivation of primary school teachers, completes the school education, rural primary school will be education line valuable education experience in field practice to students, make the school education and external education at the same time, common development and support and promote each other. Normal colleges and rural primary school is a typical system, if normal colleges only on the basis of previous experience of education teaching, or rural primary school teachers only on the basis of self demand, both cannot combine communication, normal employment, cannot adapt to the actual teaching and rural primary school recruit, do not match, retain the reality of rural teachers is difficult to solve. The training of traditional primary school teachers is 1. Introduction The basic connotation and implementation background of the rural primary school teacher training mode of "double track parallel and fixed post training" Yuanqing Yu, Yan Wang, Qifeng Zhou, Yaoyang Wei Department of Education, Huanggang Normal University, Huanggang 43800, China Corresponding author: Yan Wang (Email: 2469527001@qq.com) Abstract: "Dual-track parallel, fixed post training" rural primary school teacher training mode is to take normal colleges and rural primary schools as the main body of dual-track education, through regular tripartite meetings, improve the rural teacher enrollment, employment system and other education mechanism, to complete the teaching and education work. Through the supply and demand analysis of the current situation of rural primary school education, the disadvantages of the existing training mode are clarified. This paper will combine the general education mode of local colleges and universities, and continuously improve the construction of teachers and improve the quality of rural education from the aspects of constructing multi-party collaborative training program, optimizing the teaching system of general teachers in rural primary schools, and establishing a scientific and effective assessment and evaluation system. : "Dual-track parallel, Collaborative training", Primary school general practice teachers, Training path. Keywords: "Dual-track parallel, Collaborative training", Primary school general practice teachers, Tr unilaterally coordinated by colleges and universities, and without the establishment of a collaborative platform between colleges and rural primary schools. In the process of training primary school teachers, rural primary schools cannot participate in and cooperate in time and effectively, which ultimately leads to a certain gap between the primary school teachers trained by colleges and universities and the realistic needs of rural areas. For example, there is a phenomenon of imbalance of subject structure and single curriculum and evaluation system in college teaching, and practical problems such as single structure of teachers in rural primary schools."Dual-track parallel, collaborative training" refers to the cooperation between normal colleges and rural primary schools according to the needs of rural primary school teachers, and the targeted training of "multi-subject teachers", "general subject teachers" and "versatile teachers" based on their own ability.[1]Improving the collaborative mechanism, building a collaborative system, building a collaborative education platform between universities and rural areas, and realizing the common development of universities and rural primary schools (supply side and demand side) are the key measures to solve the mismatch and disharmony between the supply talents of colleges and universities and the demand talents of rural primary schools. International Journal of Education and Humanities ISSN: 2770-6702 \| Vol. 8, No. 3, 2023 International Journal of Education and Humanities ISSN: 2770-6702 \| Vol. 8, No. 3, 2023 Research on the Training Mode of "Dual‐track Parallel and Fixed Post Training" Yuanqing Yu, Yan Wang*, Qifeng Zhou, Yaoyang Wei 1.2. Research background of "dual-track parallel and fixed-post training" The 20th National Congress of the Communist Party of China stressed: " We should fully implement the Party's educational policy, carry out the fundamental task of cultivating morality, and train socialist builders and successors who are morally, intellec, physically, aesthetic, and hardworking. We will continue to develop people- centered education, accelerate the development of a high- quality education system, develop quality-oriented education, and promote equity in education. We will strengthen the construction of teachers' ethics, train high-quality teachers, and promote the social practices of respecting teachers and valuing education.”[2]In the face of the new development blueprint, the quality and structure of education in China have 174 been greatly improved, but the construction of rural teachers still needs to be further improved. After investigation and research, it is believed that the problem of subject structure imbalance in rural primary schools needs to be solved urgently. The balance and rationality of the subject structure of teachers play an important role in ensuring the quality of education and teaching and promoting the long-term development of the school. The inadequacy of subject structure is not conducive to cultivating students 'personalized development and improving their comprehensive quality, but also to the improvement of teachers' comprehensive quality and the development of school education and even local education. with lifelong learning development ability of high-quality professional innovative primary and secondary school teachers.”[4]Point out the direction for colleges and universities to cultivate innovative talents for rural primary schools. In 2017, Huanggang Normal University actively responded to the policy guidance, based on the current situation of rural education development and the law of physical and mental development of teenagers, formulated the talent training program for general teachers, and jointly carried out the localized training of general teachers in primary schools. By 2021, the School of Education of Huanggang Normal University has trained more than 800 primary education teachers. After graduation, they have all returned to the rural primary schools and teaching centers in their hometown to give back to the education cause in their hometown. Through the exploration of the training mode of general practice teachers in rural primary schools, Huanggang Normal University has solved the problem of general practice teacher education through reform and development, forming the Huanggang characteristics and experience of general practice teacher education in primary schools. 1.2. Research background of "dual-track parallel and fixed-post training" Rural primary schools in eastern Hubei have a large demand for general teachers, but there are few universities with the mode of education and training for general teachers. The connection between the demand of normal education in universities and rural primary schools in eastern Hubei needs to be improved, and the training path of general teachers in rural primary schools still needs to be explored. After verifying the relevant literature and the proportion of rural primary school teachers in the eastern Hubei region, it is learned that the subject structure of rural primary school teachers is unbalanced: the proportion of Chinese and mathematics teachers is more than half of the total teachers; teachers in other subjects, such as music, physical education and art, have a small proportion of the total number of teachers. The current educational situation of rural primary schools is that the supply of Chinese and math teachers generally meets the needs of rural primary schools, but there is a lack of sound, physical and beauty teachers. In order to promote the all-round development of students, teachers who were originally engaged in Chinese and mathematics have to hold the teaching task of sound, physical and beauty subjects. As a result, the phenomenon of "one division multiple subjects" and "one division multiple classes" appeared, and most subsubject teachers acted as the role of "general teachers". According to the "investigation and analysis of competent primary school curriculum", we know that rural primary school teachers have serious subjects, and sufficient teachers are competent for Chinese, mathematics and other subjects; few teachers are competent for English, music and comprehensive practice. Due to the current situation of school teaching, teachers are temporarily engaged in "general practice teaching", and have not received systematic learning and training of general practice education, and do not have enough ability to be competent for the general practice teaching position. Their knowledge level and comprehensive ability need to be further improved, and it is difficult to ensure the quality and level of teaching. p The research on the training mode of rural primary school teachers with "double track and fixed post training" plans to carry out the practice and exploration of the training mode reform of rural primary school teachers in combination with the implementation of the targeted commission training project of Huanggang Normal University. 1.2. Research background of "dual-track parallel and fixed-post training" This model aims to improve the structural shortage of rural school teachers in eastern Hubei, and strive to promote the improvement of teachers' comprehensive quality. It not only conforms to the trend of education development under the background of globalization, but also highlights the teacher training mode with Chinese characteristics. Therefore, the research of this topic is very targeted and necessary. 2.2. It is conducive to promoting the development of compulsory education and the improvement of education quality in rural areas. The collaborative education mechanism of "dual-track parallel and fixed post training" is to establish a long-term win-win cooperation mechanism with normal schools as the main training body, rural primary schools as the main participation body and local government as the undertaking point. Unimpeded three-party coordination requires coordination in the following aspects: On May 22,2017, Huanggang people's government office about the Huanggang rural primary school teachers general directional sponsored plan notice, required to orientation training way, for the city village primary school, teaching point targeted to cultivate a batch of in need of general teachers, according to the ideal faith, moral sentiment, solid knowledge, a good teacher of kindness, focus on building a "go, stay, match, teach well" localization of rural teachers, promote education precision poverty alleviation, improve the quality of rural education.[5] g p (1) Regular tripartite meetings. Through regular meetings to build a communication platform, normal colleges and rural primary schools can understand each other's needs, and clarify the responsibilities and obligations of colleges and rural primary schools. In the tripartite meeting, corresponding rules should be formulated and guiding documents should be signed to provide rules for the specific practice of collaborative education. In admissions season, graduation season key time node, through the tripartite meeting entrust training agreement, guarantee employment agreement, cooperation agreement, guarantee the school training rural primary school teachers, through the tripartite meeting, rural primary school should sponsored students academic status, moral requirements, skill level requirements and sign the corresponding documents. Under the mutual cooperation and joint supervision of the three parties, rural primary schools regularly inspect the primary school teachers trained by normal colleges, normal colleges regularly inspect the protection of the rights and interests of primary school teachers in rural primary schools, and local governments should assess and supervise the completion of the cooperation between rural primary schools and normal colleges. To sum up, the three parties composed of local government, normal colleges and rural primary schools should build a communication platform through joint meetings, supervise each other and implement them in the cooperation, and solve the problems such as the inconsistent employment of students in normal colleges and the insufficient resources of rural teachers. 2. The Research Significance of the "Double-Track Parallel, Fixed Post Training" Mode This training mode has more clear direction, and enables students to clarify their development direction —— Rural primary school general teachers, this post is of great significance to the construction of the country and society, and to the promotion of rural development, which can reflect the high personal value and social value of practitioners. 2. The Research Significance of the "Double-Track Parallel, Fixed Post Training" Mode Teachers subject structure imbalance and teachers do not have the ability of general teaching, makes the rural primary school education too much attention to students 'Chinese, mathematics discipline, but the sound body beauty course attention is insufficient, seriously affected the students' comprehensive development, for the development of quality education, promote education fair is very bad. With the continuous development of China's economic level and the great progress of education science and technology, the increasing difference between the rich and the poor in urban and rural areas has been highlighted, thus driving the inequality between urban and rural education resources and the increasing gap between urban and rural education level."Dual-track parallel, create positions training" rural primary school general teachers training mode, based on the rural primary school education teaching present situation, under the guidance of primary school general teachers training requirements, with the cultivation of combining theory with practice, to cultivate primary school general professionalism and can adapt to the position in rural teacher in this project, has an irreplaceable significance. In order to consolidate the foundation of rural education, strengthen the construction of the general practice of teachers. In the Rural Teacher Support Plan (2015-2020), important measures to expand the supplementary channels for rural teachers include encouraging local undergraduate normal universities to train "versatile" teachers in various forms according to the needs of local rural education.[3]And, on September 30,2018, the Ministry of Education issued the Ministry of Education on the implementation of outstanding teacher training plan 2.0 opinion ", clearly put forward, the future of primary school teachers will" cultivate a batch of education feelings deep, solid professional foundation, innovative teaching, good at comprehensive education and 2.1. It reflects the "people-oriented approach". It fits with the people oriented education concept That is It fits with the people-oriented education concept. That is, It fits with the people-oriented education concept. That is, 175 strength for rural basic education. to attach importance to people, understand people, respect people, care for people, and enhance and develop people's own values. The training mode of "dual-track training, fixed post training" for rural primary school teachers respects the personalized development needs of many college students who are interested in rural basic education, and stimulates the initiative and creativity of college students in learning and practice. 2.3. It is conducive to promoting the economic and cultural development in rural areas. The establishment of this training model will provide a large number of applied talents for rural primary schools, To optimize the subject structure and teaching team in rural primary schools, Make it more scientific and reasonable, The primary school teachers trained in colleges and universities can fully conform to the situation of different rural areas, For injecting more local cultural power, To help school-age children in different rural areas build awareness of the importance of learning knowledge, Form the wind of want to learn, studious, love to learn, Breaking with conventional ideas, Forming a strong educational and learning atmosphere; In accordance with the rural revitalization and development policies, Develop more possible talents for the local area, Give back to the society and return to the home in the future, Promote the construction and development of my hometown, This policy indirectly provides strength for the economic and cultural development of rural areas in China. Adhere to the humanistic principle. Embodies the fundamental purpose of serving the people wholeheartedly of the communist party of China, adhere to all to the people as the center, see the rural people's desire for education and demand, can adjust measures to local conditions, effectively resolve rural areas in the primary education demand for teachers, less available talent supply problem, but also help to resolve the rural areas of primary school teachers talent structure imbalance, for the students' comprehensive comprehensive development, to cultivate youth generation, intelligence and physique comprehensive development power, make the students have more development possibility, and help rural revitalization of our country. 2.2. It is conducive to promoting the development of compulsory education and the improvement of education quality in rural areas. q y This training mode can adjust the existing problems in the rural basic education according to local conditions, And the students trained by this model, On the one hand, with a complete professional quality, Master the scientific and advanced education and teaching methods, Be able to bring the standard subject knowledge and advanced thinking methods under the requirements of the new curriculum standards into rural primary schools, Help the students to learn more and more comprehensive knowledge, Cultivate a broad vision, Reduce the difference with urban primary education conditions and levels, Meet the requirements of equalization of education; on the other hand, During the period of choosing such a major, learning four years of professional skills and theoretical knowledge, and training to rural primary education, Have the time and opportunity to think about your adaptability to the professional direction, Students who insist on completing the training after selection can be in the training cycle, Acquisition of the system knowledge, Firmly engaged in the ideal and faith of rural education positions, Cultivate the love for rural culture and education, Form an extremely high degree of industry loyalty, Can promote the development of rural education; More with the batch of cultivation, Can provide a steady stream of (2) Improve the system of enrollment and employment of 176 to implement all the work. to implement all the work. rural teachers. Taking Huanggang City as an example, in 2017,40 general teachers in Huanggang rural primary schools were recruited in Wuxue, and students were trained in Huanggang Normal University. Before the enrollment announcement, the counties and cities of Huanggang municipal government found out the job needs, and determined the specific village and school posts according to the planned number. The education bureau of each county and city formed an enrollment publicity team to go deep into the enrollment schools. When determining employment positions, the county and city education bureau, examinees and Huanggang Normal University sign a directional training agreement to clarify the rights and obligations of both parties. After the examinee signs the agreement, the school uniformly arranges classes to study. County city education bureau to qualified examinee concentration to determine directional training work post. The job position is determined according to the principle of "score + region". 2.2. It is conducive to promoting the development of compulsory education and the improvement of education quality in rural areas. Through the coordination of the local government and the cooperation between normal colleges and rural primary schools, it is essential to provide practice time and practice positions for primary school teachers in normal colleges. Only by combining theory with practice can the trained primary school teachers adapt to the actual education situation of rural primary schools. 2.2. It is conducive to promoting the development of compulsory education and the improvement of education quality in rural areas. In the appointed PeiSheng recruitment led by Huanggang government, Huanggang normal college as the theme, rural elementary school report teachers demand involved, in the Huanggang normal college admissions deployment, illustrates the requirements for commissioned students and employment, has been clear about the normal colleges, rural primary school, commissioned students and the responsibility and obligation of local government, to contribute to the Huanggang rural primary school teachers staff shortage. In order to consolidate the achievements of poverty alleviation through education in the underdeveloped central and western regions, promote the high-quality and balanced development of education, and realize the revitalization of rural education, the Ministry of Education has put forward the "special Plan for excellent teachers". Huanggang according to the provincial education department and other nine departments of the Hubei province poverty region outstanding teachers training plan measures (Hubei teachers [2021] 3) and Huanggang city bureau of education on the good 2022 outstanding teachers training special plan related matters notice spirit, entrust Huanggang normal college directional training 100 municipal excellent teachers. In order to avoid the outstanding teachers returning to grassroots education after graduation, the students selected for the "Special Plan" need to sign employment agreements with the counties and urban areas of Huanggang City in advance. At the same time, in order to retain excellent teachers, students who participate in the "Excellent Teachers Special Program" not only exempt tuition fees, accommodation fees and subsidize living expenses during the study period, but also enjoy the official establishment of government teachers on the school.[6]In addition to Huanggang Normal University, Hubei Normal University and Hubei Second Normal University also include the "Special Plan". p (2) Establish a cooperative training mechanism between normal colleges and universities, and optimize the five-year schooling system for all teachers. Although general practice teachers do not want to have the ability to teach all subjects in primary schools, they should have the ability to organically combine subjects in primary schools. Therefore, the training requirements of general practice teachers and subsubject teachers are higher. The five-year schooling system for general practice teachers refers to 3 years of junior college + 2 years of undergraduate course. In contrast, college teachers pay more attention to skill training and vocational training, while college teachers pay more attention to the development of students' thinking and cognition. 2.2. It is conducive to promoting the development of compulsory education and the improvement of education quality in rural areas. Compared with the five- year teachers, the four-year teachers receive more comprehensive learning, and to a certain extent, the five-year teachers are more in line with the educational needs of rural primary schools. Therefore, normal colleges and universities should strengthen the contact with colleges and universities, clarify and understand their own training objectives of general practice teachers, implement phased teaching, and maximize the comprehensive development of general practice teachers' ability. In order to ensure the quality of teachers in the five- year system, the threshold should ensure the quality of students; secondly, the corresponding universities should provide good learning guarantee for students; finally, the corresponding normal colleges should pay attention to the training of teachers in the five-year colleges and universities. p y g (3) Deepen the combination of theoretical learning and educational practice of primary school teachers. The theoretical learning of primary school teachers is very important, which has a vital influence on the teacher quality and knowledge level of primary school teachers. Rural primary schools are the first line of rural education, and primary school teachers must have some practical experience before working. Through the specific education and teaching practice, the textbook knowledge is flexibly used. Therefore, primary school teachers need to use internship, practice, volunteer teaching and other ways to build a bridge for the integration of theoretical learning and practical teaching. Through the coordination of the local government and the cooperation between normal colleges and rural primary schools, it is essential to provide practice time and practice positions for primary school teachers in normal colleges. Only by combining theory with practice can the trained primary school teachers adapt to the actual education situation of rural primary schools. (3) Deepen the combination of theoretical learning and educational practice of primary school teachers. The theoretical learning of primary school teachers is very important, which has a vital influence on the teacher quality and knowledge level of primary school teachers. Rural primary schools are the first line of rural education, and primary school teachers must have some practical experience before working. Through the specific education and teaching practice, the textbook knowledge is flexibly used. Therefore, primary school teachers need to use internship, practice, volunteer teaching and other ways to build a bridge for the integration of theoretical learning and practical teaching. 3.2. Build a multi-party collaborative training scheme (1) Establish a cooperative training mechanism between the government and normal colleges, and improve the enrollment and employment system for all teachers in rural primary schools. Before the establishment of the cooperative training mechanism between the government and normal colleges, the government should first investigate the job needs of teachers in counties and cities, formulate the training plan for teachers in rural primary schools according to the survey data, and then select high-level normal colleges to undertake the training tasks. The normal colleges and universities that undertake the training task should, according to the guiding documents of the government, formulate the corresponding enrollment plan of rural primary school teachers, and integrate the high-quality teaching resources of the school to support the training of "rural primary school teachers", so as g y g (3) Establish a cooperative training mechanism between normal colleges and rural primary schools, and improve the talent training program. Talent training program has a guiding role in talent training. In order to ensure that the talent training program for general teachers has practical value and meets the educational needs of rural primary schools, normal colleges should invite rural primary schools to participate in the formulation process when formulating the talent training program for general teachers. At the same time, rural primary schools should provide practical positions for normal colleges, and build a bridge to realize the integration of theoretical 177 learning and practical teaching. learning and practical teaching. the current situation of rural primary school education and teaching, the special grade teachers of primary school and the professional teachers of normal schools can be invited to jointly compile the curriculum materials of subject and teacher education courses, so as to give full play to the respective advantages of both sides and ensure the practicability and scientificity of the teaching materials. Take the Talent Training Program for Primary Education Majors (Teachers) of Huanggang Normal University (2019 edition) as an example. The revision of the program will not only include teachers 'representatives, experts from outside the school, and students' representatives, but also representatives of the basic education industry. Moreover, the training objectives in this program conform to the basic concept of "Professional Standards for Primary School Teachers (Trial)", which promotes the comprehensive development of students majoring in primary school education. 3.4. Assessment and evaluation system (1) Reform of the assessment and evaluation system. The establishment of teaching quality testing and student evaluation system, to provide an effective guarantee for the "double track parallel, fixed post training" rural primary school teachers training mode. The teaching quality testing system includes the spot check and supervision of the Academic Affairs Office, the irregular hall tour during the classroom and self-study period, the review of teachers' courseware and test preparation materials before class, and the understanding of the implementation of the expected classroom effect. The student evaluation system includes the assessment of each subjects, the assessment of daily classroom performance and homework completion, and the written examination of mid-term and final comprehensive theoretical knowledge, among which the proportion of trial lecture, written test and daily assessment is 3:3:4. 3.2. Build a multi-party collaborative training scheme For the graduation requirements of primary school education professional students to emphasize the norms of teacher ethics and educational feelings, to become a qualified teacher, we must first love the students, love the cause of education, on this basis to strictly regulate themselves. Teachers have a role model for students, especially primary school students their mental development is not mature, the three views are still in the continuous development and improvement, primary school teachers' three views on the growth of students is crucial. Compared with cities and towns, the living and teaching conditions in rural areas are relatively difficult, and the development prospects also have certain limitations, which lead to it difficult to recruit and retain primary school teachers. Therefore, the rural primary school teachers are required to have the spirit of selfless dedication, can take root in the place where the people need, and contribute to the development of rural education. We should pay attention to the diversity of teaching materials construction forms. Normal colleges can cooperate with rural primary schools to build shared high-quality online courses according to the characteristics of the Internet era. Various teaching resources such as excellent teaching videos, case collection, teaching plans, courseware, question bank, student exercises and expert teaching videos in rural primary schools are uploaded to the online course resource database. It is convenient for students to learn and draw lessons from them more truthfully and intuitively. In recent years, affected by the epidemic, the education internship of all students in primary education has been limited to a certain extent. The construction of shared high-quality online courses can make up for this weakness to some extent, providing a guarantee for the education internship of all students in primary education. 3.3. Optimize the teaching system of all general teachers in rural primary schools (1) Rebuilding the curriculum system. Curriculum system is the core of the core of talent training. Therefore, the reconstruction of curriculum system is crucial to training general primary school teachers. General primary school teachers belong to applied talents, should focus on cultivating students' "scientific application" ability, give priority to with applied research and practical operation, need students have the ability of production and service line, so the design of the curriculum system should be subject knowledge and education teaching ability training, and the traditional subject system curriculum cannot directly reflect the education teaching ability, so need a new type of curriculum system, the system no longer depends on the knowledge classification and the nature of the curriculum, but must reflect the "crossover" this important feature, namely task driven curriculum fusion. The curriculum under this system integrates the subject knowledge, teacher ethics norms and educational feelings into the ability cultivation, and takes the education and teaching ability cultivation as the core and the subject knowledge as the support. This integration can occur either in the course or between the courses. The course is no longer a relatively independent knowledge unit, but there must be an organic connection between the courses. The curriculum system changes from a relatively independent tree structure of the course unit to the network structure of organic connection between the courses. This network makes for a shortest path between each course, making the distinction of the types or properties of the courses no longer important, and making the whole curriculum system an organic whole. (2) Research on the assessment and evaluation standards. Primary school education general professional personnel training scheme as the main reference, examine whether the trainer meet strengthen ethics, love rural education, good education quality, outstanding teaching ability, insist on lifelong learning elementary school general teacher core quality, can the theoretical knowledge applied to practice adhere to khalid ents, keep the noble moral sentiment, deep understanding of directional area rural primary school characteristics, continuous learning and reflection, become the ideal beacon of rural primary school students watch. In terms of personal basic ability, whether you have: primary school teacher qualification certificate, English level 4, Putonghua Level 2 A or above. (2) Research on the assessment and evaluation standards. References [1] Li Jingwei. The connotation, characteristics and necessity of cultivating general teachers in primary schools [J]. Education Guide, 2018 (2): 75-80. In the specific tracking of talent training results, The ability to apply what we have learned to teaching practice should be the key point to test and assess the effectiveness of talent training, Practical ability also includes education and teaching knowledge, teaching ability, research ability and other aspects, Specific information can be divided into: one, Education and teaching knowledge, Master the basic theory of pedagogy, modern educational technology, and various subject teaching methods in primary schools, Establish an advanced scientific teaching concept, The implementation of moral tree people, Implement the requirements of teacher ethics; Second, teaching ability, To specifically examine the basic skills of their teachers, That is, teaching design, blackboard writing design, three words, teaching skills; Third, the research ability, Need to actively assess whether the teachers can fully understand the students' personal characteristics, Combine pre-class preparation with after-class reflection, Establish a sense of lifelong learning, Master the effective research methods, Explore meaningful teaching cases, To promote the rural primary school education and teaching research work contribution strength. [2] Xi Jinping: Hold high the great banner of socialism with Chinese characteristics and strive for building a modern socialist country in an all-round way —— Report at the 20th National Congress of the Communist Party of China [EB / OL].http ：//www.moe.gov.cn.2022-10-25. [3] Notice of The General Office of the State Council on Printing and Issuing the Rural Teacher Support Plan (2015-2020) [EB / OL].http ：//www.moe.gov.cn.2015-06-08. [4] Opinions of the Ministry of Education on the Implementation of the Excellent Teacher Training Plan 2.0, Teacher No.13,2018 [EB / OL].http: //www.moe.gov.cn.2018-09-30. [5] Huanggang Municipal Education Bureau. The Directed Training Program for Rural Primary School Teachers in Huanggang City [EB / OL] (2017-11-19) [2022-11-16] http: / / jyj.hg.gov.cn/art/2017/11/19/art_9163_199976.html [6] Huanggang Normal University. In 2022, Huanggang Normal University "Excellent Teacher Plan" enrollment brochure [EB/OL].(2022-9-12)[2022-11-16]http: //www.hgnu.edu.cn/2022/0917/c4061a83230/page.htm [7] Hubei Enrollment information network. Introduction of Majors of Huanggang Normal University [EB / OL] (2021-06-18) [2022-11-16] http: / / zsxx.e21.cn . 3.3. Optimize the teaching system of all general teachers in rural primary schools Primary school education general professional personnel training scheme as the main reference, examine whether the trainer meet strengthen ethics, love rural education, good education quality, outstanding teaching ability, insist on lifelong learning elementary school general teacher core quality, can the theoretical knowledge applied to practice adhere to khalid ents, keep the noble moral sentiment, deep understanding of directional area rural primary school characteristics, continuous learning and reflection, become the ideal beacon of rural primary school students watch. In terms of personal basic ability, whether you have: primary school teacher qualification certificate, English level 4, Putonghua Level 2 A or above. g (3) Follow-up study on talent training effect After the trained students enter the rural primary school, carry out the tracking survey with every six-month cycle, and take the following four factors as comprehensive assessment criteria: first, after the training, whether the students can fully adapt to the rural primary school environment and the learning level of local students; second, whether the knowledge reserve is perfect, can extract the application, whether the knowledge can effectively output helps students to learn; third, whether meet the requirements of the (3) Optimize the construction of teaching materials. Textmaterials are crucial in the process of teacher teaching and students' learning. In order to promote the coordinated development of the curriculum content of general practice and 178 professional quality; fourth, professional loyalty and working attitude, can adapt to the current work, and still maintain the enthusiasm for education and teaching. Acknowledgment Hubei Provincial Department of Education Philosophy and Social Sciences Research Project in 2021.Project Title: Research on the Training Model of "Double-track Parallel and Fixed-post Cultivation" for Rural Primary School Teachers (Project No. 21Y237) [8] The Ministry of Education issued opinions on Strengthening the Educational Practice of Normal University Students. Teachers No.2 [EB / OL] http: / / www.moe.gov.cn.2016-03- 21. 179
https://openalex.org/W2110629875	https://portal.research.lu.se/files/1568666/5463870.pdf	English	null	Downregulation of Type II Diabetes Mellitus and Maturity Onset Diabetes of Young Pathways in Human Pancreatic Islets from Hyperglycemic Donors	Journal of diabetes research	2,014	cc-by	6,897	Link to publication Citation for published version (APA): Taneera, J., Storm, P., & Groop, L. (2014). Downregulation of Type II Diabetes Mellitus and Maturity Onset Diabetes of Young Pathways in Human Pancreatic Islets from Hyperglycemic Donors. Journal of Diabetes Research, 2014, Article 237535. https://doi.org/10.1155/2014/237535 Citation for published version (APA): Taneera, J., Storm, P., & Groop, L. (2014). Downregulation of Type II Diabetes Mellitus and Maturity Onset Diabetes of Young Pathways in Human Pancreatic Islets from Hyperglycemic Donors. Journal of Diabetes Research, 2014, Article 237535. https://doi.org/10.1155/2014/237535 Total number of authors: 3 Downregulation of Type II Diabetes Mellitus and Maturity Onset Diabetes of Young Pathways in Human Pancreatic Islets from Hyperglycemic Donors. Taneera, Jalal; Storm, Petter; Groop, Leif Published in: Journal of Diabetes Research Published in: Journal of Diabetes Research Citation for published version (APA): Taneera, J., Storm, P., & Groop, L. (2014). Downregulation of Type II Diabetes Mellitus and Maturity Onset Diabetes of Young Pathways in Human Pancreatic Islets from Hyperglycemic Donors. 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LUND UNIVERSITY PO Box 117 221 00 Lund +46 46-222 00 00 Hindawi Publishing Corporation Journal of Diabetes Research Volume 2014, Article ID 237535, 7 pages http://dx.doi.org/10.1155/2014/237535 Hindawi Publishing Corporation Journal of Diabetes Research Volume 2014, Article ID 237535, 7 pages http://dx.doi.org/10.1155/2014/237535 Jalal Taneera, Petter Storm, and Leif Groop Department of Clinical Sciences, Diabetes & Endocrinology, Lund University Diabetes Center, Sk˚ane University Hospital, Lund University, 20502 Malm¨o, Sweden Department of Clinical Sciences, Diabetes & Endocrinology, Lund University Diabetes Center, Sk˚ane University Hospital, Lund University, 20502 Malm¨o, Sweden Correspondence should be addressed to Jalal Taneera; jalal.taneera@med.lu.se Received 22 July 2014; Revised 22 September 2014; Accepted 25 September 2014; Published 14 October 2014 Academic Editor: Daisuke Koya Copyright © 2014 Jalal Taneera et al. This is an open access article distributed under the Creative Commons A which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is p Although several molecular pathways have been linked to type 2 diabetes (T2D) pathogenesis, it is uncertain which pathway has the most implication on the disease. Changes in the expression of an entire pathway might be more important for disease pathogenesis than changes in the expression of individual genes. To identify the molecular alterations in T2D, DNA microarrays of human pancreatic islets from donors with hyperglycemia (𝑛= 20) and normoglycemia (𝑛= 58) were subjected to Gene Set Enrichment Analysis (GSEA). About 178 KEGG pathways were investigated for gene expression changes between hyperglycemic donors compared to normoglycemic. Pathway enrichment analysis showed that type II diabetes mellitus (T2DM) and maturity onset diabetes of the young (MODY) pathways are downregulated in hyperglycemic donors, while proteasome and spliceosome pathways are upregulated. The mean centroid of gene expression of T2DM and MODY pathways was shown to be associated positively with insulin secretion and negatively with HbA1c level. To conclude, downregulation of T2DM and MODY pathways is involved in islet function and might be involved in T2D. Also, the study demonstrates that gene expression profiles from pancreatic islets can reveal some of the biological processes related to regulation of glucose hemostats and diabetes pathogenesis. 1. Introduction from individuals with specific diseases. For example, mRNA expression profiles are generated from thousands of genes from samples of one of two classes such as cancer [9]. The differential expressed genes between classes can be ranked based on their differentiation. However, the remaining chal- lenge is how to interpret a given list of genes into biological mechanism. T2D is a multifactorial disease characterized by increased blood glucose level due to both a defect in insulin secretion from pancreatic beta-cells and impaired insulin action at the target cells. The disease is estimated to affect more than 350 million people in 2020 worldwide (http://www.idf.org/ diabetesatlas/) and to contribute to other diseases such as atherosclerotic vascular disease, blindness, and kidney failure [1]. Mootha et al. have developed a statistical methodology called Gene Set Enrichment Analysis (GSEA) to define whether a given gene set is significantly enriched in a list of genes ranked by their correlation with a phenotype of interest [8, 10]. GSEA has been shown to have an increased capacity to detect modest but coordinated changes in prespecified set of related genes. GSEA has been successfully used to uncover altered metabolic pathways in several applications such as human diabetic muscle [8], comparing mouse models of can- cer with human tumors using gene-expression profiling [11], lung cancer [10], characterization of acute megakaryoblastic Several molecular pathways have been implicated in the disease process: insulin receptor signalling [2], carbohydrate metabolism [3], ER stress related pathway [4], cytokine signalling [5], exocytosis [6], and oxidative phosphorylation [7, 8]. However, it is unclear which of these or other pathways are disturbed in and might be responsible for T2D in its common form. DNA microarrays expression analysis enables scientists to investigate the altered transcript levels in particular tissue Journal of Diabetes Research 2 2 Table 1: Characteristics of human pancreatic donors. Normoglycemic Hyperglycemic 𝑁(male/female) 58 (34/24) 20 (11/9) Age (years) 60.9 ± 10.9 64 ± 8.9 BMI 25.4 ± 2.9 28.5 ± 4.5 HbA1c 5.4 ± 0.3 6.9 ± 1.0 Purity 70 ± 16 63 ± 20 Donors with diabetes 0 10 Data represented as mean ± SD. Table 1: Characteristics of human pancreatic donors. leukemia [12], and interaction between mRNA and miRNA in HIV-mediated neurodegeneration [13] and comparing whole blood gene expression profiling from lean and obese individuals [14]. 2. Materials and Methods 2.1. Human Pancreatic Islets. Islets from cadaver donors (78 donors) were provided by the Nordic Islet Transplan- tation Program (www.nordicislets.org), Uppsala University. All procedures were approved by the ethics committees at Uppsala and Lund Universities. Islets were obtained from 68 nondiabetic donors (30 females, 37 males, age 59 ± 10, BMI 25.9 ± 3.5, HbA1c 5.5 ± 1.1, and days of culture 3.5 ± 1.9) and 10 T2D donors (4 females, 6 males, age 60.7 ± 12, BMI 28.1±4.5, HbA1c 7.1±1.2, and days of culture 2±0.9). Purity of the islet preparations was assessed by dithizone staining, insulin content, and contribution of exocrine and endocrine tissue as previously described [15]. The islets were cultured in CMRL 1066 (ICN Biomedicals, Costa Mesa, CA, USA) supplemented with 10 mM/L HEPES, 2 mM/L l-glutamine, 50 𝜇g/mL gentamicin, 0.25 𝜇g/mL Fungizone (GIBCO, BRL, Gaithersburg, MD, USA), 20 𝜇g/mL ciprofloxacin (Bayer Healthcare, Leverkusen, Germany), and 10 mM/L nicoti- namide at 37∘C (5% CO2) prior to RNA preparation. 2.4. Gene Set Enrichment Analysis (GSEA). The GSEA soft- ware tool (version 2.0.13, www.broadinstitute.org/gsea/) was used to identify KEGG pathways (MSigDB, version 4.0) that show an overrepresentation of up- or downregulated genes between donors with hyperglycemia (HbA1c > 6%, 𝑁= 20) and normoglycemia (HbA1c < 6%, 𝑁= 30). Briefly, an enrichment score was calculated for each gene set (i.e., KEGG pathway) by ranking each gene by their expression difference using Kolmogorov-Smirnov statistic, computing a cumulative sum of each ranked in each gene set, and recording the maximum deviation from zero as the enrichment score. 2.2. Microarray Gene Expression in Human Pancreatic Islets. RNA was isolated with the AllPrep DNA/RNA Mini Kit (Qiagen, Hilden, Germany). RNA quality and concentration were measured using an Agilent 2100 bioanalyzer (Bio- Rad, Hercules, CA, USA) and Nanodrop ND-1000 equip- ment (NanoDrop Technologies, Wilmington, DE, USA). The microarrays (GeneChip Human Gene 1.0 ST) were performed using the Affymetrix standard protocol as previ- ously described [15]. The array data were summarized and normalized with robust multiarray analysis (RMA) method using the oligo package from BioConductor. Also, batch correction was done with COMBACT function from SVA package from BioConductor. All data are MIAME compliant, and the raw data have been deposited in a MIAME database (GEO, accession number: GSE 50398 and GSE 50397). 2.5. Statistical Analysis. Data are presented as means ± S.D. Differences in expression levels were analyzed by Student’s 𝑡- test or nonparametric Mann-Whitney tests. 1. Introduction Here, we employed GSEA to determine whether the 178 selected KEGG pathways are altered between islet gene expression from donors with normoglycemia and hyper- glycemia. Pathway enrichment analysis showed that MODY and T2DM pathways are downregulated in hyperglycemic islets. The mean centroid of gene expression of T2DM and MODY pathways was shown to be significantly associated with insulin secretion and HbA1c level, which highlight that these pathways are involved in islet function. solution and was treated with 95% O2-5% CO2 to obtain constant pH and oxygenation. After preincubation, the buffer was changed to a KRB buffer containing either 1 mM (basal secretion) or 16.7 mM glucose (stimulated secretion). The islets were then incubated for 1 h at 37∘C in a metabolic shaker (30 cycles per min). Immediately after incubation, an aliquot of the medium was removed for analysis of insulin using a radioimmunoassay kit (Euro-Diagnostica, Malm¨o, Sweden). Insulin content in homogenized human islets was assessed by ELISA (Mercodia, Uppsala, Sweden) and values were normalized to the total DNA in each sample as determined by a fluorometric assay (Quant-iT PicoGreen, Invitrogen Molecular Probes, Stockholm, Sweden). 2. Materials and Methods Correlation tests were analyzed using nonparametric Spearman’s tests. The mean centroid represents the normalized gene expression levels of all genes from all individuals in the analysis with a mean of 0 and a variance of 1. All statistical tests were per- formed using the Statistical Package for the Social Sciences (SPSS) version 19.0 software (SPSS, Chicago, IL, USA). 4. Discussion Although several pathways have been implicated in T2D pathogenesis, most of these studies were performed in non- pancreatic tissues. In this study, we used human pancreatic islets obtained from 78 donors. Each donated pancreatic islet was systematically characterized by performing cDNA microarray in addition to measuring insulin response to glucose and glycemic status (HbA1c). Next, we examined individual expression value of the 45 genes in the T2DM and the 24 genes in MODY pathway. We found that 14 out of the 45 genes of the T2DM pathway (31%) and 9 out of the 24 genes of MODY (37.5%) contributed significantly to core enrichment whose expression was lower in hyperglycemic than in normoglycemic donors (Figures 2(a)-2(b)). Also, expression of the 23 genes was signifi- cantly reduced in diabetic compared to nondiabetic donors (Table S1 in Supplementary Material available online at http://dx.doi.org/10.1155/2014/237535). Insulin receptor sub- strate genes (IRS4) were shown to be low/not expressed in human pancreatic islets (Figure 2(a)). Glucokinase (GCK), solute carrier family 2 (facilitated glucose transporter), member 2 (SLC2A2), pancreatic and duodenal homeobox 1 (PDX1), and v-maf musculoaponeurotic fibrosarcoma onco- gene homolog A (MAFA) overlapped between core enrich- ment of the two pathways. Interestingly, plotting the mean centroid of the 14 and 9 downregulated genes in T2DM and MODY pathways showed a positive correlation with insulin secretion and negative correlation with an HbA1c level (Figures 2(c)-2(d)) suggesting that these pathways are involved in regulation of insulin secretion and glycemic status. Mean centroid of the 31 and 55 upregulated genes in the core enrichment of proteasome and spliceosome pathways showed no correlation with insulin secretion but positively correlated with HbA1c level (Figures 2(e)-2(f)). We also anal- ysed the real score of expression and differential expression of g g y Our data presented additional evidence into the biologi- cal processes that differentially were regulated in pancreatic islets from normoglycemic and hyperglycemic donors. The downregulated pathways (T2DM and MODY) in hyper- glycemic donors were due to decreased expression of several protein-encoded genes, which indicate a reduction in protein synthesis in pancreatic islets. Recently, Del Rosario et al. reported that regions in promoter of genes involved in T2DM and MODY pathways are more likely to be differentially methylated between diabetic and nondiabetic donors com- pared to other genes [16]. 3. Results 2.3. Glucose-StimulatedInsulinSecretion. Isletswerehand-picked under a stereomicroscope and preincubated for 30 min at 37∘C in Krebs Ringer bicarbonate (KRB) buffer (pH 7.4) con- taining (in mM) 120 NaCl, 25 NaHCO3, 4.7 KCl, 1.2 MgSO4, 2.5 CaCl2, 1.2 KH2PO, 10 HEPES supplemented with 0.1% bovine serum albumin, N-2 hydroxyethylpiperazine-N󸀠-2- ethanesulfonic acid (10 mmol/L), and 1 mmol/L glucose. Each incubation vial contained 12 islets in 1.0 mL KRB buffer In this study, we used human islet microarray expression data obtained from 78 donors. The donors were subdivided into 58 normoglycemic donors with HbA1c level <6% and 20 hyperglycemic donors with HbA1c level >6% (Table 1). Normalized expression microarray data were subjected to pathway analysis of GSEA algorithm using 178 Kyoto Ency- clopedia for Genes and Genomes (KEGG) pathways. Pathway Journal of Diabetes Research 3 Table 2: List of down- and upregulated pathways in hyperglycemic donors. Table 2: List of down- and upregulated pathways in hyperglycemic donors. Size NES NOM 𝑃value FDR 𝑞value Downregulated KEGG pathways KEGG TYPE II DIABETES MELLITUS 45 −1,898 0 0,077 KEGG MATURITY ONSET DIABETES OF THE YOUNG 24 −1,812 0 0,108 KEGG OOCYTE MEIOSIS 108 −1,543 0,02 0,485 KEGG PROGESTERONE MEDIATED OOCYTE MATURATION 83 −1,453 0,04 0,70 KEGG SNARE INTERACTIONS IN VESICULAR TRANSPORT 38 −1,583 0,05 0,610 Upregulated KEGG pathways KEGG PROTEASOME 44 2,026 0,005 0,030 KEGG SPLICEOSOME 125 1,898 0,01 0,075 KEGG DNA REPLICATION 36 1,621 0,04 0,821 KEGG PRIMARY IMMUNODEFICIENCY 35 1,604 0,02 0,71 KEGG CYTOKINE CYTOKINE RECEPTOR INTERACTION 251 1,593 0,003 0,619 KEGG GLYOXYLATE AND DICARBOXYLATE METABOLISM 16 1,529 0,03 0,614 Ranking of the genes set was done using GSEA 2.0.13. NES: normalized enrichment score; NOM: nominal; FDR: false discovery rate. genes in the core enrichment of proteasome and spliceosome pathways in normoglycemic versus hyperglycemic and in nondiabetic versus diabetic donors (Tables S2 and S3). enrichments were evaluated by their normalized enrichment score (NES), nominal 𝑃value, and false discovery rates (FDR). GSEA identified 4 pathways (𝑃 < 0.05) which were downregulated in the hyperglycemic islets compared to normoglycemic islets (Table 2). At FDR < 25%, only two pathways (T2DM and MODY) were shown to be significant (Table 2 and Figures 1(a)-1(b)). On the other hand, six enriched pathways were upregulated in the hyperglycemic donors (𝑃< 0.05), while at FDR < 25% only two pathways (proteasome and spliceosome) were significant (Table 2 and Figures 1(c)-1(d)). 4. Discussion In this study, the islet expression mean centroid of downregulated genes in T2DM and MODY pathways correlated positively with insulin secretion and negatively with HbA1c level, suggesting that appropriate expression of these genes is required for sufficient insulin secretion and glucose homeostasis. Notably, most of the downregulated genes have been implicated in diabetes pathogenesis; SLC2A2 (Glut2) is involved in 𝛽-cell function and insulin secretion [17]. Mice lacking SLC2A2 showed early diabetes and abnormal postna- tal pancreatic islet development [18]. ABCC8 is a regulator of ATP-sensitive K(+) channels and insulin release. A mutation in ABCC8 was observed in patients with hyperinsulinemic hypoglycemia of infancy [19] and associated with T2D [20]. KCNJ11, together with ABCC8, regulates transmembrane potential and thereby glucose-stimulated insulin secretion 4 Journal of Diabetes Research 0 0.00 0.25 0.50 0.0 2500 5000 7500 10000 12500 15000 17500 20000 −0.6 −0.25 −0.5 −0.4 −0.3 −0.2 −0.1 Enrichment score (ES) Enrichment profile Ranking metric scores Rank in ordered dataset Zero cross at 9293 Ranked list metric (Signal2Noise) Hits “DIA” (positively correlated) “NONDIA” (negatively correlated) Enrichment plot: KEGG TYPE II DIABETES MELLITUS (a) 0.00 0.25 0.50 −0.25 0.0 −0.6 −0.7 −0.5 −0.4 −0.3 −0.2 −0.1 Enrichment score (ES) 0 2500 5000 7500 10000 12500 15000 17500 20000 Rank in ordered dataset Zero cross at 9293 Ranked list metric (Signal2Noise) “DIA” (positively correlated) “NONDIA” (negatively correlated) Enrichment profile Ranking metric scores Hits KEGG MATURITY ONSET DIABETES OF THE YOUNG Enrichment plot: (b) 0.5 0.4 0.3 0.2 0.1 0.0 0 2500 5000 7500 10000 12500 15000 17500 20000 Enrichment score (ES) Rank in ordered dataset 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment profile Ranking metric scores Hits Enrichment plot: KEGG SPLICEOSOME (c) 0 2500 5000 7500 10000 12500 15000 17500 20000 Rank in ordered dataset Enrichment profile Ranking metric scores Hits 0.5 0.6 0.4 0.3 0.2 0.1 0.0 Enrichment score (ES) 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment plot: KEGG PROTEASOME (d) Figure 1: GSEA plot. The analysis was performed against the KEGG database for differential enriched pathways between hyperglycemic and normoglycemic islets. Enrichment plots for the downregulated pathways are shown in graphs (a) and (b) and upregulated pathways are shown in graphs (b) and (c). 4. Discussion The 𝑦-axis represents the value of the ranking metric; the 𝑥-axis represents the rank for all genes. Bottom: plot of the ranked list of all genes. Top: the enrichment score for the gene set as the analysis walks along the ranked list. The score at the peak of the plot is the enrichment score (ES) for this gene set and those genes appearing before or at the peak are defined as core enrichment genes in this gene set. Lower levels of expression are represented in shades of blue and higher expression is represented in red. 0.00 0.25 0.50 −0.25 0.0 −0.6 −0.7 −0.5 −0.4 −0.3 −0.2 −0.1 Enrichment score (ES) 0 2500 5000 7500 10000 12500 15000 17500 20000 Rank in ordered dataset Zero cross at 9293 Ranked list metric (Signal2Noise) “DIA” (positively correlated) “NONDIA” (negatively correlated) KEGG MATURITY ONSET DIABETES OF THE YOUNG Enrichment plot: 0 0.00 0.25 0.50 0.0 2500 5000 7500 10000 12500 15000 17500 20000 −0.6 −0.25 −0.5 −0.4 −0.3 −0.2 −0.1 Enrichment score (ES) Rank in ordered dataset Zero cross at 9293 Ranked list metric (Signal2Noise) “DIA” (positively correlated) “NONDIA” (negatively correlated) Enrichment plot: KEGG TYPE II DIABETES MELLITUS Enrichment plot: KEGG TYPE II DIABETES MELLITUS Enrichment score (ES) Ranked list metric (Signal2Noise) Rank in ordered dataset Rank in ordered dataset Enrichment profile Ranking metric scores Hits (a) Enrichment plot: KEGG SPLICEOSOME Enrichment profile Ranking metric scores Hits (a) Enrichment profile Ranking metric scores Hits (b) 0.5 0.4 0.3 0.2 0.1 0.0 0 2500 5000 7500 10000 12500 15000 17500 20000 Enrichment score (ES) Rank in ordered dataset 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment profile Ranking metric scores Hits Enrichment plot: KEGG SPLICEOSOME (c) 0 2500 5000 7500 10000 12500 15000 17500 20000 Rank in ordered dataset Enrichment profile Ranking metric scores Hits 0.5 0.6 0.4 0.3 0.2 0.1 0.0 Enrichment score (ES) 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment plot: KEGG PROTEASOME (d) Figure 1: GSEA plot. The analysis was performed against the KEGG database for differential enriched pathways between hyperglycemic and normoglycemic islets. Enrichment plots for the downregulated pathways are shown in graphs (a) and (b) and upregulated pathways are shown in graphs (b) and (c). 4. Discussion The 𝑦-axis represents the value of the ranking metric; the 𝑥-axis represents the rank for all genes. Bottom: plot of the ranked list of all genes. Top: the enrichment score for the gene set as the analysis walks along the ranked list. The score at the peak of the plot is the enrichment score (ES) for this gene set and those genes appearing before or at the peak are defined as core enrichment genes in this gene set. Lower levels of expression are represented in shades of blue and higher expression is represented in red. (a) 0.5 0.4 0.3 0.2 0.1 0.0 0 2500 5000 7500 10000 12500 15000 17500 20000 Enrichment score (ES) Rank in ordered dataset 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment plot: KEGG SPLICEOSOME Enrichment plot: KEGG PROTEASOME 0 2500 5000 7500 10000 12500 15000 17500 20000 Rank in ordered dataset 0.5 0.6 0.4 0.3 0.2 0.1 0.0 Enrichment score (ES) 0.00 0.25 0.50 −0.25 Ranked list metric (Signal2Noise) “DIA” (positively correlated) Zero cross at 9293 “NONDIA” (negatively correlated) Enrichment plot: KEGG PROTEASOME Enrichment plot: KEGG SPLICEOSOME En Ranked list metric (Signal2Noise) Ranked list metric (Signal2Noise) Rank in ordered dataset Rank in ordered dataset Figure 1: GSEA plot. The analysis was performed against the KEGG database for differential enriched pathways between hyperglycemic and normoglycemic islets. Enrichment plots for the downregulated pathways are shown in graphs (a) and (b) and upregulated pathways are shown in graphs (b) and (c). The 𝑦-axis represents the value of the ranking metric; the 𝑥-axis represents the rank for all genes. Bottom: plot of the ranked list of all genes. Top: the enrichment score for the gene set as the analysis walks along the ranked list. The score at the peak of the plot is the enrichment score (ES) for this gene set and those genes appearing before or at the peak are defined as core enrichment genes in this gene set. Lower levels of expression are represented in shades of blue and higher expression is represented in red. 4. Discussion (b) 0 1 2 3 4 5 6 0 0.5 1 Maturity onset diabetes of young mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1c (%) −0.5 −1.5 −1 (d) ( ) 0 1 2 3 4 5 6 0 0.2 0.4 0.6 0.8 1 Type II diabetes mellitus mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1 (%) −1.2 −1 −0.8 −0.6 −0.4 −0.2 y y g (d) 0 1 2 3 4 5 6 0 0.1 0.2 0.3 0.4 Spliceosome mean centroid Insulin release 0 2 4 6 8 10 12 HbA1c (%) −0.2 −0.3 −0.1 (f) (c) 0 1 2 3 4 5 6 0 0.1 0.2 0.3 Proteasome mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1c (%) −0.3 −0.2 −0.1 (e) (d) (c) (f) (e) Figure 2: Genes differentially expressed in T2DM and MODY pathways. Gene expression analysis of genes in the T2DM and MODY pathway showed that 14 out of the 45 genes of the T2DM (a) pathway (𝑃< 0.05) and 9 out of 24 genes of MODY (b) (𝑃< 0.05) have lower expression in hyperglycaemic compared to normoglycemic donors. (c) Correlation of mean centroid of the 14 downregulated genes in T2DM pathway showed positive correlation with insulin secretion (𝑅= 0.33; 𝑃= 0.01) and negative correlation with HbA1c level (𝑅= −0.57; 𝑃= 0.00001). (d) Correlation of mean centroid of the 9 downregulated genes in MODY pathway showed positive correlation with insulin secretion (𝑅= 0.31; 𝑃= 0.01) and negative correlation with HbA1c level (𝑅= −0.59; 𝑃= 0.000006). (e) Correlation of mean centroid of the 31 upregulated genes in proteasome pathway showed no correlation with insulin secretion (𝑅= −0.05; 𝑃= 0.7) and positive correlation with HbA1c level (𝑅= 0.3; 𝑃= 0.03). (f) Correlation of mean centroid of the 55 upregulated genes in spliceosome pathway showed no correlation with insulin secretion (𝑅= 0.16; 𝑃= 0.23) and positive correlation with HbA1c level (𝑅= 0.26; 𝑃= 0.06). insulin gene expression [23]. Defects in this gene caused maturity onset diabetes of the young type 4 (MODY4) [24]. PAX6, point mutations in the PAX6 gene shown to disrupt islet morphology and decreased numbers of 𝛽, 𝛼, and PP cells [25]. Also, a mutation in the gene has been shown to cause early-onset diabetes [26]. MAFA is required for islet beta-cell differentiation and activates the insulin and in pancreatic beta-cells. 4. Discussion 5 5 Journal of Diabetes Research 0 100 200 300 400 Normoglycemic Hyperglycemic 500 1000 1500 2000 Normalized mean expression IRS4 IRS2 ABCC8 CACNA1A CACNA1C CACNA1D GCK KCNJ11 MAFA MAPK10 PDX1 PIK3R3 SLC2A2 CACNA1B (a) 0 500 1000 Normoglycemic Hyperglycemic 6000 13000 Normalized mean expression PAX6 IAPP NEUROD1 NKX2-2 NKX6-1 GCK MAFA PDX1 SLC2A2 (b) 0 1 2 3 4 5 6 0 0.2 0.4 0.6 0.8 1 Type II diabetes mellitus mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1c (%) −1.2 −1 −0.8 −0.6 −0.4 −0.2 (c) 0 1 2 3 4 5 6 0 0.5 1 Maturity onset diabetes of young mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1c (%) −0.5 −1.5 −1 (d) 0 1 2 3 4 5 6 0 0.1 0.2 0.3 Proteasome mean centroid Insulin secretion 0 2 4 6 8 10 12 HbA1c (%) −0.3 −0.2 −0.1 (e) 0 1 2 3 4 5 6 0 0.1 0.2 0.3 0.4 Spliceosome mean centroid Insulin release 0 2 4 6 8 10 12 HbA1c (%) −0.2 −0.3 −0.1 (f) Figure 2: Genes differentially expressed in T2DM and MODY pathways. Gene expression analysis of genes in the T2DM and MODY pathway showed that 14 out of the 45 genes of the T2DM (a) pathway (𝑃< 0.05) and 9 out of 24 genes of MODY (b) (𝑃< 0.05) have lower expression in hyperglycaemic compared to normoglycemic donors. (c) Correlation of mean centroid of the 14 downregulated genes in T2DM pathway showed positive correlation with insulin secretion (𝑅= 0.33; 𝑃= 0.01) and negative correlation with HbA1c level (𝑅= −0.57; 𝑃= 0.00001). (d) Correlation of mean centroid of the 9 downregulated genes in MODY pathway showed positive correlation with insulin secretion (𝑅= 0.31; 𝑃= 0.01) and negative correlation with HbA1c level (𝑅= −0.59; 𝑃= 0.000006). (e) Correlation of mean centroid of the 31 upregulated genes in proteasome pathway showed no correlation with insulin secretion (𝑅= −0.05; 𝑃= 0.7) and positive correlation with HbA1c level (𝑅= 0.3; 𝑃= 0.03). (f) Correlation of mean centroid of the 55 upregulated genes in spliceosome pathway showed no correlation with insulin secretion (𝑅= 0.16; 𝑃= 0.23) and positive correlation with HbA1c level (𝑅= 0.26; 𝑃= 0.06). References Proteasomes are protein complexes with a main function to regulate and degrade unnecessary or damaged proteins by proteolysis, while spliceosome is a complex molecular machine assembled from snRNPs and protein complexes. Splicing is a known process when spliceosome removes introns from a transcribed pre-mRNA. Both of the protea- some and the spliceosome pathways showed upregulation in hyperglycemic donors. The findings are potentially very important as there are several lines of evidence which reported glucose or hyperglycemia to influence proteasome and splicing. Recent studies have shown that high glucose and diabetes affect proteasome activity [32, 33]. The link of hyperglycemia to proteasome raises several questions such as how hyperglycemia can modulate proteasome targeting and activity and whether this modulation occurs in a cell-specific manner. Hribal et al. reported that chronic hyperglycemia impairs insulin secretion by affecting splicing in RIN 𝛽-cell line and human islet [34]. Osmark et al. reported pronounced tissue-specific differences in the splicing of TCF7L2 with forms containing exons 4 and 15 being the most abundant in islets. The incorporation of exon 4 in islets was shown to correlate positively with HbA1c levels [35]. Also, a short TCF7L2 mRNA variant in subcutaneous fat is associated with hyperglycemia and impaired insulin action in adipose tissue [36]. Although these reports do not prove causality, they suggest an effect of plasma glucose levels on splicing.hi [1] P. 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PDX1 is involved in the early development of the pancreas and plays a major role in glucose-dependent regulation of 6 Journal of Diabetes Research 6 glucagon promoters [27]. MAFA functions as a downstream mediator of PAX6 in regulating the specification of insulin and glucagon expressing cells [28]. NEUROD1 is reported to regulate expression of the insulin gene [29], and mutations in this gene result in maturity onset diabetes of the young type 6 (MODY6) [30]. FOXA2 is involved in glucose homeostasis and regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis [31]. Hence, these data suggest that the downregulation of the 23 genes in T2DM and MODY pathways is a causative for insulin secretion impairment. collaborative Project Grant Dnr. 521-2008-2974, strategic research area Grant (EXODIAB: Dnr. 2009-1039), and Lin- naeus Grant (LUDC): Dnr. 349-2008-6589). Human pancre- atic islets were provided by the Nordic Network for Clinical Islet Transplantation by the courtesy of O. Korsgren, Uppsala, Sweden, supported by EXODIAB and grants from JDRF. The authors thank SCIBLU Genomics at Lund University for their help with microarrays preparation. Acknowledgments [13] L. Zhou, G. M. Pupo, P. Gupta et al., “A parallel genome- wide mRNA and microRNA profiling of the frontal cortex of HIV patients with and without HIV-associated dementia shows This work was supported by grants from the Swedish Research Council incl.Project Grant (LG: Dnr. 251-2007-4037, Journal of Diabetes Research 7 [28] I. Artner, B. Blanchi, J. C. Raum et al., “MafB is required for islet 𝛽cell maturation,” Proceedings of the National Academy of Sciences of the United States of America, vol. 104, no. 10, pp. 3853– 3858, 2007. the role of axon guidance and downstream pathways in HIV- mediated neurodegeneration,” BMC Genomics, vol. 13, no. 1, article 677, 2012. [14] V. Skov, S. 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https://openalex.org/W2943536380	https://journal.unnes.ac.id/nju/index.php/jpii/article/download/16564/9028	English	null	Motivation in Information Communication and Technology-Based Science Learning in Tamil Schools	Jurnal Pendidikan IPA Indonesia/JPPI : Jurnal Perndidikan IPA Indonesia	2,019	cc-by	8,846	Jurnal Pendidikan IPA Indonesia http://journal.unnes.ac.id/index.php/jpii http://journal.unnes.ac.id/index.php/jpii Correspondence Address E-mail: akalai.vkam@gmail.com ABSTRACT The purpose of this study is to examine the impact of Information Communication Technology (ICT) utilisa tion in learning science to improve students’ motivation. This study examined 100 students from Malaysian Tamil vernacular schools to learn ICT-based science.Research design used a mixed method approach involving quantitative and qualitative data. Students answered the Science Motivation Questionaire II (SMQ II) instru ment based on the dichotomous scale that uses “Yes or No” questions. Open-ended questions were also analysed qualitatively. The instrument was divided into five categories, namely Intrinsic Motivation, Grade Motivation, Self-efficacy Motivation, Self-determination and ICT Motivation. The findings showed that all 25 items of SMQ IIhad significantly high level of motivation. 90.2% of respondents agreed that ICT-based learning of Science stimulated their Intrinsic Motivation. Respondents admitted that factors such as Elements of ICT (46.6%), As sisted Learning (30.10%) and Stimulated Interest (23.3%) had been their attraction towards the usage of ICT in learning Science. This study also identified that respondents regarded the level of internet accessibility (34.7%) and health (25.5%) as being the main obstacles to using ICT during Science learning. Findings from this study show health issues hinder the usage of ICT in learning Science. Therefore this new element should be taken into account so as not to become a constraint for their motivation. The significance of this study is to enable Tamil vernacular school students to show interest in studying science. © 2019 Science Education Study Program FMIPA UNNES Semarang © 2019 Science Education Study Program FMIPA UNNES Semarang Keywords: science learning, Tamil vernacular school students, ICT DOI: 10.15294/jpii.v8i1.16564 Accepted: November 16th, 2018. Approved: March 25th, 2018. Published: March 28th, 2019 Accepted: November 16th, 2018. Approved: March 25th, 2018. Published: March 28th, 2019 Keywords: science learning, Tamil vernacular school students, ICT K. Shanmugam1 and B. Balakrishnan2 1,2Sultan Idris Education University, Malaysia K. Shanmugam1 and B. Balakrishnan2 JPII 8 (1) (2019) 141-152 JPII 8 (1) (2019) 141-152 Jurnal Pendidikan IPA Indonesia © 2019 Science Education Study Program FMIPA UNNES Semarang K. Shanmugam1 and B. Balakrishnan2 1,2Sultan Idris Education University, Malaysia INTRODUCTION Chernobilsky, 2012; Ciampa, 2014; Fook & Sid hu, 2013; Granito & Chernobilsky, 2012; Ziden et al., 2011) find that ICT is capable of facilitating teaching presentation, diversification in teaching activities, making lessons more fun and inter esting as well as improving student motivation. Furthermore, the use of ICT is identified as the best measure to confront the complexity of the science subject (Sin et al., 2013). In the digital world today, the position of the computer and Information Communi cation Technology (ICT), remains on the front row.The integration of ICT in education is also growing rapidly. This means ICT can be an ef fective and interesting instructional medium of Teaching and facilitating (T&F) in education, especially in Science.The rapid development of ICT has become one of the attractive factors in enhancing the motivation of students (Granito & ICT and its motivation becomes an effec tive channel to improve the achievement of pu pils in Science T&F. The goal of the Curriculum Development Division for Science subjects is to K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 142 and Tamil schools were placed in the rubber plan tation estates. “inculcate the interest and to develop the crea tivity of pupils (Malaysia, 2012). In reality the capacity and the impact of ICT is not known in general (Granito & Chernobilsky, 2012). Accor ding to Naim & Tunggak (2014), teachers and parents need to realise that motivating students to excel in academics at school is a priority. This is proven when the target in science education Per kara 4.9, Dasar Pendidikan Negara (Abu Bakar et al., 2017) based on the ratio of 60 per cent of science stream students and 40 per cent of small letter arts streem students has yet to be achieved. In fact this trend of depreciation is increasingly worrying. Currently, the ratio is at 21:79 (Abu Ba kar et al., 2017). The interest of vernacular schools was given attention in the Razak Report 1956. This report emphasises the importance of education system through the use of mother tongue. Accor ding to the Malaysia Commissioner of Law Re vision. (1996) Act 550, ‘ National Primary Type Schools’ refers to government primary schools or primary schools with government assistance that use Mandarin or Tamil language as the medium of instruction. INTRODUCTION The rationale behind the setting up of vernacular schools is to avoid racial issues in a country that has a variety of native languages and different lifestyles. Vernacular schools came about without the existence of a clear uniformity on the syllabus, guidelines, and standards. Cur rently, the building and facilities status of Tamil schools are experiencing changes that are encou raging, but the achievement of academic perfor mance has not reached the achievement as seen in Chinese Vernacular schools. The level and standard of Tamil Vernacular Schools is expected to be well established with the application of ICT in Teaching and Facilitation (T&F). The effecti veness of ICT can be proven by the existence of studies in the use of ICT in Science T&F. In addition to this scenario, the achieve ment in Sekolah Jenis Kebangsaan Tamil (SJK (T))Ujian Pencapaian Sekolah Rendah (UPSR) Science in Tamil vernacular schools in Perak, a state in Malaysia, has been on the decline since 2011 (Perak State Education Department,2017). MIB Cooperation (2017), expect that the gap between National Schools, Tamil Vernacular Schools, and Chinese Vernacular Schools can be reduced.However, the problem of self-motivati on among students of Tamil vernacular schools and the decline in the achievement in Science is an agenda that should be given attention. Tamil vernacular schools were initially formed with various deficiencies without the existence of a clear consistency on the syllabus, guidelines and standards. Now, changes and development in the nation’s education policies have brought about changes to the Tamil Vernacular School educa tion system in Malaysia. At present, Tamil Ver nacular Schools operate based on a standard syl labus, guidelines, textbooks, trained teachers and the pupils of year 6 will be sitting for the UPSR public examination. The syllabus has been the ba sis for the acceptance of science among students. The Science syllabus and Kurikulum Standard Sekolah Rendah (KSSR) science content in Ta mil Vernacularschools is designed in accordance withthe National Education Philosophy where the knowledge of science and technology skills can be mastered. (Malaysia, 2012). ICT includes “one software to convert, store, protect, process, transfer and acquire in formation” regardless of place and time. In line with the development of the world, Malaysia also applies technology in education which is experi encing constant changes. The development of a country is only meaningful when it is driven by mastery in science and technology (Abdullahi, 2014). INTRODUCTION This discovery indi cates a digital chasm which is an ongoing issue in rural areas. Science education has become an agenda in the world arena. As the world is becoming globally closer and is dependable on intellectual resources, the question of how to achieve high scientific literacy for all children around the world becomes a priority for us to consider. Knowledge of science in a globalised world plays an impor tant role in supporting and reinforcing economic and cultural globalisation (Chiu & Duit, 2011). Evaluation of Educational Achievement (IEA) on the other hand organises international bench- marking for the Science subject in order to enhan ce the performance of Science in the internatio nal arena (BERNAMA, 2018). In line with this international bench-marking, Thomas & Watters (2015), also admit that Science is the most power ful way in thinking and understanding in order to address the various problems that are shaking the world.According to Chiu &Duit (2011), Germa ny organised a symposium on standards in scien ce education where it invites representatives from across the country to contribute their expertise and experience on the development of scientific standards in various cultures . Menon et al. (2012) found the use of Wiki pedia in learning Science also carries some adver se effects such as involving long periods of time for search and discovery of materials based on links. There are also students who identifytheir lack of proficiency in English to be the main ob stacle for them to learn online. In addition, the extensive syllabus becomes a hindrance in the use of ICT. As a suggestion ‘The structured and constricted syllabus in the science curriculum has to be changed to meet the demands of virtu al learning in line with the required 21st century skills (Menon et al., 2012). The findings by Yasak et al. (2010), sho wed that the frequency of ICT use is moderate among Science teachers in Johor, a state in Ma laysia, but the use of software in science educati on has helped the learning process and aroused a sense of fun among the students. Furthermore, students who are creative and innovative live in a cheerful atmosphere (Yasak et al., 2010). This situation becomes a reality with the presence of various elements such as graphics, animation and audio which are more interesting compared to textbooks. INTRODUCTION In a world moving towards digital media, the role of ICT in education has become inc reasingly important. Students who do not mas ter ICT, are considered as outdated witha lack of knowledge and poor academic achievement (Ahmad, 2016). This period is known as the in formation era with many communication chan nels such as email, SMS, internet browsing, mes senger, web-meeting, viber, BBM, Google Talk, Skype,Web 2.0 Technologysuch asYoutube, Blog, Twitter dan Facebook (Atan, 2016). Now, vario us online software applications are designed with the aim to facilitate T&F on special topics for all levels (Garba et al., 2015).This situation has cre ated space and broader opportunities for teachers to use ICT in their T&F. Moreover, ICT and the digital age has changed the way pupils learn and access information (Shivana et al., 2014). After the arrival of the British, the edu cation system in Malaya developed more syste matically.Vernacular schools were established in accordance with the Labour Act (1900). Tamil schools had features that reflected the different ethnic groups in the Indian community. Each of the ethnic groups sent their children to get educa tion in schools that used their respective mother tongue.The British established these schools in accordance to the location of Indian settlements, Ministry of Education Malaysia has laun ched various strategies to encourage teachers to use ICT in class (Ahmad, 2014). However, its use K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 143 (2013), suggested that every school have a coor dinator to handle ICT. He also proposed thatmo dern computer labs be made available in schools. Science teachers should be given the opportunity to become ICT literate while in service. Ministry of Education Malaysia has provided all ICT fa cilities and given exposure to teachers under the platform of smart school. However the level of acceptance and the usage by teachers is still the main issue. The technical aspects of ICT usage are identified as key issues and becomes a stigma that inhibits the success of ICT and Frog VLE (Hamzah & Yeop, 2016). is still not widespread. Integrating ICT in the clas sroom is not as easy as it might be. The findings by Mohamed et al. (2012), show the percentage of computer ownership and the use of ICT in ru ral areas is low while on the other hand basic ICT skills are at a medium level. INTRODUCTION Yıldız & Aktaş (2015) emphasise the use of the computer as a medium of teaching in a constructive learning environment that en courages students to have more positive attitudes and motivation towards teaching. Garba et al. (2015) reported that Malaysia is at a more advanced level in their efforts to rea lise the true potential of ICT in education. Ma laysia has access to internet connection at home, at work, at school, and in public places. ICT te aching materials allow the materials to be used repeatedly: they can be used actively wherever the pupils are without a time limit. This situation mo tivates the teaching content data which can be de livered more easily through the use of ICT(Noor AiniAhmad, 2014). ICT factors also contribute to an inc rease in student motivation. The rapidly gro wing ICT has become one of the major factors in improving student motivation, according toGranito&Chernobilsky (2012) and (Ananiadou & Rizza et al. 2010). According to Ziden et al. (2011) ICT has enhanced the motivation to learn more effectively and with quality. Ciampa (2014), Fook & Gurnam (2013), Granito & Chernobilsky (2012) find that ICT is capable of facilitating the presentation of instruction, diversification of acti vities and to make teaching and learning sessions more fun and attractive to students. Furthermore, Chua & Don (2013), found that computer-based testing mode has attracted the respondent moti vation to participate in the study. Science-based ICT T&F in Malaysia (in volving 10,000 schools) is progressing into a Vir tual Learning Environment (Frog VLE). It allows pupils to be exposed where they can access a wi der content, which is exciting and interactive. The use of Frog VLE allows teachers and pupils to ac cess the learning of science no matter where they are without a time limit (Blueprint, 2016).The implementation of Frog VLE, has given space to ICT to act as a bridge to connect the urban and rural areas. Frog VLE network serves as the basis for the construction of the virtual learning plat form that can be used by teachers, students, and parents to share learning resources, implement interactive learning, and virtual communication (Ministry of Education Malaysia, 2013). Further more the use of ICT can be identified as the best measure to confront the complexity of the scien ce subject (Sin & Norishah, 2013). INTRODUCTION In an effort to instill a Science culture based on ICT, Smart Schools have been established. Thus, the ability to pursue and master the knowledge of science can strengthen Malaysia’s position in the world arena. To achieve this agenda, the use of ICT in Science subjects need to be improved. This will motivate students to explore science. ICT helps to motivate students and at the same time increase students’ performance (Sán chez-garcía et al., 2013). Each student has diffe rent needs. ICT fulfills the needs of the individual student and also helps them in their studies by motivating them to study. With these, students learn more effectively (Kler, 2015). The use of computers and mobile equip ment have turned into a new channel and is a rat her new T&F culture in instructional media in the classroom (Nikou & Economides, 2016). This is because computer technology and mobile equip ment can facilitate learning ‘anytime and anyw here’ (Nikou & Economides, 2016) and (Ahmad Rafaai et al., 2016). The ability to educate any where makesit an increasingly popular medium. Indirectly, this uniqueness has increased the int rinsic and extrinsic motivation and the efficiency of the students, Merino & López (2014) dan Ci ampa (2014).This new and contemporary media has also increased student motivation among tho se who are mediocre and weak, to learn science. Motivation in education refers to a set of beliefs about the behaviour that guides both te achers and students in a social environment be cause they interact with each other during T&F (Abu Bakar et al., 2017). According to Naim & Tunggak (2014), teachers and parents need to motivate students to achieve academic excellen ce at school which is a priority. Technological equipment and ICT have tremendously influen ced the motivation of students and this will lead to changes. The 21st century generation who are so routine are the genuine users in the world of digital technology, will understand its use much easier. In such a situation, technological equip ment will tremendously influence the motivation of students in T&F activities (Granito & Cher Halili et al. (2011) found that a person’s learning motivation has also been increased by the existence of video conferencing technology. INTRODUCTION The lack of competency of teachers and equipment caused the deterioration in the per formance of science in Tanzania (Kafyulilo et al., 2015). Cross & Board (2014) find that pupils (aged 10 to 14 years with the change in indivi dual thinking from simple to a more complex level) showed no interest and a negative attitude towards inquiring about new findings in science. Based on these findings, it is clear that ICT has a role in improving the performance of science ab road. Even so, the decline in science achievement can be attributed to the lack of ICT usage. Findings by Phang et al. (2014), found out the use of various forms of ICT in T&F were implemented in the science education in this country. In the implementation stage, the suita bility in the use of ICT, particularly in science education, is very dependable on infrastructure equipment for the implementation of T&F using ICT, creativity and teacher’s innovation and technical support staff, willingness and the inte rest of teachers and students to carry out T&F using ICT, the school administration using ICT in management (Phang et al., 2014). The study found that the levels of interest, willingness and ability of science teachers in using ICT in T&F are positive. Numerous research in the use of ICT by science teachers have been conducted in fo reign countries. However,the findings cannot be adapted according to the culture and context in Malaysia. Hence,there is a need for an intensified study in Malaysia (Ahmad, 2014). This scenario clearly indicates that research on the use of ICT among teachers should be prioritised by educa tional researchers. In ICT, Power Point is an easy way to use as a content delivery tool and is an ele ment of assistance for the elaboration of complex science concepts (Ahmad, 2014). Chua & Don The study also shows that the integration of ICT in education sow the factors of childhood development in a more comprehensive way. Ac K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 144 cording to Abdullahi (2014), the application of ICT in Science subjects, showed more success compared to other subjects. nobilsky, 2012). Yıldız & Aktaş (2015) emphasise the use of the computer as a medium of teaching in a constructive learning environment that en courages students to have more positive attitudes and motivation towards teaching. nobilsky, 2012). METHODS psychological needs of students are met, then the level of their motivation to use assessment enhancement based on mobileequipment inc reases. When student autonomy and self-rule is supported and there is a positive social influence, students will feel more motivated. Tamil vernacular schools have pupils who study all subjects in their native language which is Tamil. Science is also taught in Tamil language except for the existing Dual Language Program me. The researcher surveyed the T&L Science using ICT and what is encouraging and inhibiting ICT usage during T&L Science. The extensive addition of various low-cost mobile equipment can be obtained with wireless internet service being the main driver in this new era of education (Nikou & Economides, 2016). Research design used a mixed method approach involving quantitative and qualitative data. Quantitative research questions in this stu dy inquire about the relationships among ICT usage in T&L Science with studentmotivation. The qualitative data (open-ended question) is to make descriptive assertions about the student in using ICT during T&L Science lesson in class. In addition, students who have high intrin sic motivation show a lot of passion to learn and understand.The use of mobile technology also gives ample room for individuals to practise the culture of cooperation. Mobile technology can be a tool to convey instructions and functions as a medium to work together. Students can learn in their own way, hand in hand with others and offer advice to each other through a variety of applica tions (Ciampa, 2014). For the present study, the relationship bet ween student experiences in usage of ICT during T&L Science by the teacher and student motiva tion was examined through a reliable instrument, the Science Motivation Questionnaire II (SMQ II). SMQ II by Glynn et al. (2009) comprises 25 items (each subscale has five items) used for mo nitoring: intrinsic motivation, ICT motivation, self-determination, self-efficacy and grade moti vation. Menon et al. (2012) that examines the role of wikipedia in learning Science, found that vir tual learning guided by steps or process may inc rease the motivation of students studying science. In addition to improving motivation, such equip ment is also found to increase the camaraderie between students, lifelong learning and create a conducive learning environment. Data from the questionnaire were measu red quantitatively with a Dichotomous scale. The response pattern followed ‘Yes’ or ‘No’. K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 145 INTRODUCTION With the use of audio and video, students are motivated for being able to see and hear around which raises their level of understanding.This suggests that the use of ICT does not only attract the interest of the students to follow the process of T&F better, but helps to increase the under standing and to make it convenient for teachers to deliver their lessons (Halili et al., 2011). Students’motivation levels change and the affects are based on several factors. When the METHODS Dichotomic items are best suited for ten-year-old pupils. reading because they involved short answers writ ten by Year 4 students. For the content validation of data, the researcher used an expert from the language field (27 years’ experience) to confirm the usage of the theme and sub-theme. The Social Learning Theory was pionee red by Albert Bandura. Bandura introduced a learning process called ‘Observational Learning’ in 1986. The latest Bandura theory includes a dy namic system that describes human adaptation, learning and motivation (Woolfolk, 2010). Ac cording to Bandura, impersonation takes place through observations of the behaviour of the mo del in the person being imitated. In conclusion, imitation or modeling behaviour is the result of reinforcement.The culture of this model begins in school when students see teachers as a force to be respected. Teachers are also models of pupils because of the areas of expertise they have. The expertise in delivering knowledge based on cur rent needs such as ICT divisors is highly regarded by students. Hence, students begin to show inte rest and motivation towards learning. Indirectly, culture imitates teachers as the latest expertise encourages motivation to learn among students. Among the implications associated with T&L is that as a teacher, it is important for teachers to give each student an opportunity to observe and emulate various types of ICT-based models in order to motivate their learning.Teachers can perform activities such as giving quizzes based on latest technologies such as ICT, which chal lenge students’ minds so that their cognitive skills can grow continuously. The ability to imitate and self-efficacy help individuals to progress towards excellence. In this study, the use of ICT as one of the strategies that becomes the attraction and imperative will be examined. There were two open-ended questions that needed to be answered briefly by students. The researcher has made Back-to-Back Translation for SMQ II which was originally in English. The se SMQI I items were translated into Malay and then translated back to English with the help of an English language expert to ensure the validity of the instrument. Originally, SMQ had 30 ques tions which were reduced to 25 in SMQ II. METHODS The goal of dichotomous scale was to measure students’ interest in the usage of ICT by the teacher du ring T&L Science in class. The researcher used percentage to determine the level of the students’ motivation. The qualitative analysis was also done to the attractive factors and factors which become an obstacle during the usage of ICT in T&L Science. Fook & Sidhu (2013) find that ICT has in creased the motivation to learn more effectively and with quality. Fook & Sidhu (2013) and Ciam pa (2014) find ICT is capable to facilitate teach ing, diversification of activities to make teaching sessions and learning more fun and attractive to students. However, the findings by Granito & Cher nobilsky (2012) show different results.His fin dings show that, if given a choice, students would wish to complete their tasks using paper-pencil method compared to using the computer. Lack of interest in computers because of the ‘difficult to operate’ mentality by the user, effectuate such conclusion. This study involved random sampling that was selected based on rural areas which have an average achievement in the UPSR examination. The criteria of the pupils were based on Standard Four pupils who were currently studying in rural Tamil schools. This study involved 10 Tamil ver nacular schools that were homogeneous in the aspect of age, school infrastructure and trained instructors. The students examined in the aspect of ICT towards T&L Science based on SMQ II developed by Glynn et al. (2009). All schools of which achievement is moderate were chosen based on the median scale (Idris, 2013). The re searcher chose 10 students (from each school) to The findings of this study recommend that children need to be exposed to the use of compu ters so they can be acquainted with the techno logy at a very young age. In addition to the idea of Granito & Chernobilsky (2012) and Abubakar (2015), described though, that using technology tools will not guarantee the success of the stu dents, the students’ skills and the ability of the pupils to compete globally. K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 146 answer the questionnaires and to examine their motivation towards ICT usage. This study invol ved questionnaires (SMQII). SMQII instruments use dichotomous elements. Dichotomy items in volve short responses items such as “yes” or “no” to enable respondents to choose one of the two options given. RESULTS AND DISCUSSION This study involved 100 respondents comprising primary school students. Gender ana lysis data shows a total of 53 percent of the res pondents are female and a total of 47 per cent are male students. All the respondents were pupils at age 10 and were studying at the Tamil vernacular school located in rural Perak in Malaysia. This study examined the level of learning motivation of science based on ICT. Figure 2 shows the level of motivation of each SMQ II component that shows a high percentage, theintrinsic motivation (90.2%) being the most notable element among the pupils who learned Science based on ICT. Intrinsic motivation is followed by self efficiency motivation (87.2%), grade motivation (87%), ICT motivation (83.4%), and self determination moti vation (67.4%) respectively. y Dimensions of Self Efficiency as a whole, recorded 87.2% for all items. Items I believe (95%) I am confident (95%), ‘I’m certain’ (94%) ‘able to master’ (89%) and ‘do the best in the science lab’ (63%) show a strong passion among the students in learning Science based on ICT. Self Determi nation Motivation recorded a high percentage be cause the atmosphere in the classroom has led to this motivation. In the classroom, pupils are usu ally surrounded by teachers and friends. Teachers will know the name of every pupil, check the dai ly attendance, to correct the books and become the driving force. In such situations, the students indirectly have had such motivation (Woolfolk, 2010). This decision is in line with findings from Schumm & Bogner (2016), which show the Self efficiency on recording high score. The Study of Merino & López (2014) and Ciampa (2014) also found that indirect uniqueness that can be used to educate anywhere has increased the internal motivation and self efficiency motivation of the students. Figure 2. Motivation towards Learning Science Using ICT Figure 2. Motivation towards Learning Science Using ICT The intrinsic motivation of pupils affected the learning of Science based on ICT. The fin ding shows the use of ICT in Science T&F mo tivate the research decision that shows: interest (98%), fun (96%), curious about the latest inven tion (94%), related to life (88%) and a more mea ningful life (75%). All these items show the stages of acceptance of Intrinsic Motivation which are high among the respondents. METHODS Figure 1. Research Model In addition, the intrinsic motivation of pri mary school students who like to compete with their classmates are successful in evoking their internal motivations. Therefore every student wants to achieve the ‘accessible maximum level’ based on each and everyone’s unique potential (Sri, 2013; Abdullah & Hendon, 2016). This study aimed to figure out how teach ers’ ability and usage of ICT’s impact on stu dent motivation to learn Science in rural Tamil Schools. This research model will give an input to answer the Research Questions. However, these results differ from the study conducted by Schumm & Bogner (2016), which examines Science Motivated teenagers. The study found that the mean score Grade indi cates a higher level compared to the mean score of Intrinsic Motivation. The Grade motivation in this study is at a high level but the motivation is in the third place after Internal Motivation and Self Efficiency Motivation. METHODS For this study, the researcher took SMQ II with 25 questions but integrated it with ICT components and dropped components related to career moti vation because they are not suitable for primary school students and more suitable for teenagers who will leave school. Previous studies such as Nikou & Economides (2016), Schumm & Bog ner (2016), Chua & Don (2013), and Glynn et al. (2011) has observed university respondents, teenagers and secondary school students. Ho wever, for this study, the respondents were only primary school students. Thus, career motivation was dropped for this study.The researcher used percentage to determine the level of the students’ motivation. The qualitative analysis was also done for the attractive factors and factors which become an obstacle during the usage of ICT in T&L Science. The first Research Question was analysed using percentage. The answers were analysed ma nually by counting the percentage.The objective of this research is to examine the level of moti vation among Tamil Vernacular school students. The motivation level for these primary school students was measured using the dichotomous scale whichonly involves two options “yes” or “no” as answers. For primary school children of 10 yearsold, it is a good way to analyse and it is an appropriate way to examine the level of their motivation. Bandura’s (1986) Social Cognitive Theory is one of the most influential theories in psycho social and education field, especially applicable to understanding an individual’s innovation of adoption in terms of their belief and attitude development (Leong, 2017). Bandura’s (1986) Social Cognitive Theory emphasises on social learning through modelling. Students in this stu dy who observed teachers adopting a particular innovation (ICT usage in T&L Science) may be more inclined to consider adoption themselves. By observing teachers using ICT, students acqui re skill and knowledge, beliefs and attitudes. The research Model for the study as shown in figure below. For the second research question, data were analysed using the manual procedure. The mes and sub-themes were coded using manual K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 147 Figure 1 Research Model of ICT in educating anywhere is capable of evo king the intrinsic motivation of the pupils. Hami mah & Buerah (2014) suggested that the intrin sic motivation of the pupils can be enhanced by providing learning experiences that are fun with teachers increasing the number of activities or science assignments involving ICT. RESULTS AND DISCUSSION They also support Science T&F ful ly which involves investigation and practical trai ning based on ICT. This decision is also aligned with the theory of Bandura (1986) where delive ring current knowledge skillfully such as infusing ICT in learning Science is highly regarded by students. Thus, the students begin to show inte rest and are motivated towards learning science. Indirectly, the teacher is imitating culture because the latest technology expertise motivates pupils to learn among themselves. ICT motivation has recorded 83.4% which is also thought to be high. This entry is considered relevant because the 21st century generation has been completely exposed to the world of techno logy. Information can be delivered and dissemi nated widely using the tools of modern techno logy. Smart phones that have internet network on its own are able to become learning aids that are capable of displaying dozens of search results. Accessibility has made this technology preferred by Education 4.0 century students where creativi ty and being innovative are emphasised. The outcomes of this study showed that audio elements have contributed to student mo tivation (7.5%) towards ICT-based Science Lear ning. This is similar to the findings by Yasak et al.(2010) which found audio-based science edu cation software attract students to study science. A total of 30.1% of the respondents admitted that they liked to use ICT because ICT helped in their learning. Issues such as ‘simple’ (13.7%), ‘informative’ (9.6%) and ‘assist learning’ (3.4%) became the main reasons for respondents. On the part of the students, ICT and the digital age has changed the way students learn and how they access information (Shivana et al., 2014). Kler (2015) stated that ICT in T&F has changed the entire concept of education and has proven to be of great benefit to students. Furthermore, the use of ICT is identified as the best measure to con front the complexity of science subject (Sin et al., 2013). Another aspect that was reviewed in this study are the factors that find the use of ICT an attraction in the classroom. The respondents have given a number of factors that have attracted them to use ICT. Figure 3 shows the factors that contribute to why ICT use in Science T&Fis at tractive in the classroom. Figure 3. RESULTS AND DISCUSSION The result of the finding is in accordance with the decision of Me rino& López (2014), Ciampa (2014), and Nikou & Economides (2016) who claim that the ability The Grade Motivational dimension shows that pupils are tied to grades that should be ob tained. 4 Grade Motivational items show high motivation. Item with ‘an A’ (96%) ‘earned a good Science grade which is important’ (96%) ‘I like to do better than other students’ (95%) and ‘get a high score’ (93%) are often involving mo tivation that are associated with grades. Overall, Grade Motivation Dimension has registered 87% Grade-based achievement is important for national exam-oriented students in Malaysia. K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 148 Ziden et al. (2011) also showed that audio and video help enhance motivation and thus improve performance. They also support Science T&F ful ly which involves investigation and practical trai ning based on ICT. This decision is also aligned with the theory of Bandura (1986) where delive ring current knowledge skillfully such as infusing ICT in learning Science is highly regarded by students. Thus, the students begin to show inte rest and are motivated towards learning science. Indirectly, the teacher is imitating culture because the latest technology expertise motivates pupils to learn among themselves. Upper primary students in primary schools are bound by this government public examination called UPSR.This achievement is considered to be important so that they are qualified to go to the secondary schools and enrolled into Form 1 (without going through Remove classes for one year). This constraint causes them to be motiva ted based on grades. The findings are similar to essential Grade Motivation similar to the studies by (Schumm & Bogner, 2016; Glynn et al., 2011). Upper primary students in primary schools are bound by this government public examination called UPSR.This achievement is considered to be important so that they are qualified to go to the secondary schools and enrolled into Form 1 (without going through Remove classes for one year). This constraint causes them to be motiva ted based on grades. The findings are similar to essential Grade Motivation similar to the studies by (Schumm & Bogner, 2016; Glynn et al., 2011). Ziden et al. (2011) also showed that audio and video help enhance motivation and thus improve performance. RESULTS AND DISCUSSION Attraction Factors of Teaching Science Using ICT A total of 23.3% of the respondents ad mitted that ICT has stimulated their interest to learn science. Elements such as ‘interest’ (5.5%), ‘active’ (4.1%) ‘thought’ (4.1%) and ‘interes ting’(3.4%) have become the main reasons to sti mulate learning. This suggests that the use of ICT media not only attracts students to follow the pro cess of T&F better, but helps to increase the un derstanding of students and makes it convenient for teachers to deliver their lessons (Halili et al., 2011). Fook & Gurnam (2013) share the same opinion in this study where ICT is found to build self-confidence, encourage creative thinking and make learning sessions more fun and interesting for students. Figure 3. Attraction Factors of Teaching Science Using ICT Figure 3 shows the attraction factors of ICT during Science T&F class that are catego rised as helpful in learning, stimulates interest and ICT elements.‘Elements of ICT’ (46.6%) is‘followed by assisted learning’ (30.1%) and ‘sti mulating the interest’ (23.3%) respectively. Elements of ICTinvolves 46.6% of pupils who are interested in using ICT. Elements such as being ‘colourful’ (25.3%), ‘audio’ (7.5%), ‘video’ (4.8%) and ‘beautiful’ (4.8%) have been the main stimulus for students to be interested in ICT use during science T&F. Such factors should be addressed by edu cators in classroom teaching. These factors will indirectly ensure that students are more focused in learning Science and, at the same time, able to improve their academic performance. Teachers need to find the best way to equip themselves with technological knowledge in line with the develop ment of ICT and be able to provide constructive The output of this study is similar to the study made by Sidek & Hashim (2016) who found that pupils’ motivation improve with the use of student-centered video clips. Finding by K. Shanmugam and B. Balakrishnan / JPII 8 (1) (2019) 141-152 149 net usage caused various health problems for them. The respondents argued that if the use of ICT continued for a certain period of time they wouldhave sore eyes, headaches and eye glare. This state is referred to as digital pressure against the eye. Proactive measures need to be taken to ensure these problems do not become a continuo us barrier which will finally inhibit the use of ICT among the younger generation. Obstacles in the Usage of ICT During Science T&F This investigation also aims to find the fac tors of barrier in the use of ICT in Science T&F. Figure4 shows the results of the study with re gards to the obstacles in the usage of ICT during Science T&F in class. Figure 4. Barriers in Teaching Science Using ICT The results also showed the disruption of tools (7.1%) has become a barrier in the current usage of ICT during Science T&F. Research by Garba et al., (2015) reported that Malaysia is at a more advanced level in its efforts to maximise the potential of ICT in education. Malaysians have access to internet connection at home, at work, at school, and in public places. Under the plat form of the creating smart schools, ICT is provi ded to schools to ensure success in the process of technological integration in the learning process (Hamzah & Yeop, 2016). The study shows fin dings that are contrary to the findings by Garba et al. (2015) and Hamzah & Yeop (2016). The issue of internet barrier (34.7%) computer restrictions at home (9.2%), computer restrictions at school (4.1%) achieved 48% (almost half of the respon dents) indicating this issue remains a primary is sue in the country in the adaptation practice of ICT in Science T&F. This research is supported by the findings of Mohamed et al. (2012) which show the percentage of computer ownership and the use of ICT in rural areas are low. This dis covery reflects the digital chasm which is an on going issue in rural areas. The government is pay ing attention to this ongoing issue. For the sake of addressing this issue (4G access is 74.88% but the speed is only 14.83 Mbps) Malaysia has imple mented Universal Service Programme (USB). The Malaysian Communications and Multime dia Commission (MCMC), under USB, serves to identify locations in rural areas with low in ternet coverage (BERNAMA, 2018). The effort and concern by the government in overcoming the digital chasm between urban and rural areas should be ongoing. Figure 4. Barriers in Teaching Science Using ICT Figure 4 shows the findings that there are various obstacles in the current use of ICT in Science T&F among students. The level of ‘in ternet accessibility’recorded 34.7% followed by ‘health constraints’ which recorded 25.5%. RESULTS AND DISCUSSION Various parties such as governments, non-governmental organi sations, parents and teachers can work actively so that this problem does not persist and should be addressed immediately in order to produce a competent young generation in the use of ICT and excel in the international arena. input among students. Teachers need to employ the use of technology during the preparation and delivery in the classroom. The digital generation is definitely interested and motivated to be more actively involved in Science T&F. CONCLUSION In this study, Intrinsic Motivation has become the dominant component of the ICT- based Science education. Elements of ICT have become attractive factors while internet acces sibility and health problems prohibit the use of ICT in Science T&F. Motivation improvement will increase achievement level in Science among students and will realise the aspirations of the go vernment based on MIB Cooperation (2017) that would reduce the gap in the percentage of pas ses in UPSR subjects between Tamil Vernacular Schools, Chinese Vernacular School and National Schools in the next ten years to come. The ratio of science to art stream students which has fallen to 21:79 can also be improved with the availabi lity of ICT-based motivation insertion in Scien ce T&F. The ability to increase the involvement of students in Science stream will be increased. Following the increase in students’ achievement, the country is also proud to join Education 4.0. Education 4.0 is compliant to the aspirations of modern technology that would certainly aggran dise Science based ICT T&F. 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Mengintegrasi Teknologi Ko munikasi dan Maklumat Sebagai Media Pen gajaran Bahasa dalam Kalangan Kanak-Kanak Awal Umur. Jurnal Pendidikan Bahasa Mela yu, 4(2), 36-43. Obstacles in the Usage of ICT During Science T&F Only a small handful of respondents said factors such as ‘time’ (10.2%),‘inaccessibility of computer at home’ (9.2%), ‘family’ (8.2%),‘tools’ (7.1%), ‘school’ (4.1%) and ‘adaptation’ (1%) became a barrier in the use of ICT during the learning of Science. The main obstacle in ICT usage is in terms of restraints of the internet. As much as 34.7% of respondents stated that they did not have the internet access at home. Even if there is internetaccess, the coverage is weak and can not help in Science T&F at home or in school. Fook & Gurnam (2013) found just as much as 47% of the respondents agreed that they have an Internet connection at home. For this study, a to tal of 34.7% of respondents claimed not to have an internet connection at home. Fook & Gurnam (2013) observed that non-functioning computer softwares become an obstacle in the use of ICT during the learning process. For the sake of st rengthening the commitment towards learning of T&F Science based on ICT, each respondent has to have a computer and an Internet connection at home. This study also identified that time restric tions involved 10.2% of the respondents. Respon dents said that theywould spend a lot of time in using ICT to search for information. Similarly, Menon et al. (2012) also found the use of Wikipe The second obstacle factor involves health. 25.5% of the respondents claimed that inter K. Shanmugam and B. 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https://openalex.org/W4281618532	https://ok.abvpress.ru/jour/article/download/535/388	Russian	null	Risk factors for urgent complications of colorectal cancer	Tazovaâ hirurgiâ i onkologiâ	2,022	cc-by	6,334	К о н т а к т ы : Светлана Николаевна Щаева shaeva30@mail.ru На развитие перфорации больше влияли стадия заболевания – местный статус опухоли T4 (ОШ 3,74; 95 % ДИ 2,61–5,48; p <0,001) и наличие стадии N+ (ОШ 1,61; 95 % ДИ 1,33–2,01; p <0,001), высокая степень злокачественности опухоли G3–G4 (ОШ 3,56; 95 % ДИ 2,08–4,93; p <0,001) и коморбидность – наличие сахарного диабета, корригируемого инсулино- терапией (ОШ 2,11; 95 % ДИ 1,78–2,42; p <0,001). Результаты. Большее влияние на развитие декомпенсированной кишечной непроходимости оказывали стадия заболевания – местный статус опухоли T4 (отношение шансов (ОШ) 3,19; 95 % доверительный интервал (ДИ) 1,66–7,12; p <0,001), высокая степень злокачественности опухоли G3–G4 (ОШ 2,93; 95 % ДИ 0,89–3,97; p = 0,008) и коморбидность – наличие конкурирующих заболеваний (ОШ 2,03; 95 % ДИ 1,84–2,39; p <0,001). На развитие перфорации больше влияли стадия заболевания – местный статус опухоли T4 (ОШ 3,74; 95 % ДИ 2,61–5,48; p <0,001) и наличие стадии N+ (ОШ 1,61; 95 % ДИ 1,33–2,01; p <0,001), высокая степень злокачественности опухоли G3–G4 (ОШ 3,56; 95 % ДИ 2,08–4,93; p <0,001) и коморбидность – наличие сахарного диабета, корригируемого инсулино- терапией (ОШ 2,11; 95 % ДИ 1,78–2,42; p <0,001). Выводы. Пациенты старческого возраста с III и IV стадиями колоректального рака и при наличии высокой комор- бидности с большей вероятностью могут иметь ургентные осложнения в виде декомпенсированной острой кишеч- ной непроходимости и перфорации опухоли. Ключевые слова: ургентные осложнения колоректального рака, декомпенсированная острая кишечная непрохо- димость, перфорация опухоли Для цитирования: Щаева С. Н. Оценка факторов риска, влияющих на развитие ургентных осложнений колорек- тального рака. Тазовая хирургия и онкология 2022;12(2):28–35. DOI: 10.17650/2686‑9594‑2022‑12‑2‑28‑35. О р и г и н а л ь н о е и с с л е д о в а н и е DOI: 10.17650/2686‑9594‑2022‑12‑2‑28‑35 4.0 О р и г и н а л ь н о е и с с л е д о в а н и е Оценка факторов риска, влияющих на развитие ургентных осложнений колоректального рака ФГБОУ ВО «Смоленский государственный медицинский университет» Минздрава России; Россия, 214019 Смоленск, ул. Крупской, 28 О р и К о н т а к т ы : Светлана Николаевна Щаева shaeva30@mail.ru Цель исследования – изучить основные факторы, оказывающие влияние на развитие ургентных осложнений колоректального рака. Svetlana Nikolaevna Shchaeva shaeva30@mail.ru Svetlana Nikolaevna Shchaeva shaeva30@mail.ru Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 К о н т а к т ы : Светлана Николаевна Щаева shaeva30@mail.ru К о н т а к т ы : Светлана Николаевна Щаева shaeva30@mail.ru Цель исследования – изучить основные факторы, оказывающие влияние на развитие ургентных осложнений колоректального рака. Материалы и методы. В ретроспективное когортное исследование включали больных, страдающих колоректальным раком и перенесших экстренные хирургические вмешательства в период с января 2016 г. по январь 2020 г. в об- щехирургических стационарах г. Смоленска. Всего было включено 214 пациентов, медиана возраста пациентов составила 66 лет. Были проанализированы социально-демографические характеристики (пол, возраст, место проживания (городская или сельская местность), образование, трудовая занятость, профессиональный и семейный статусы), анамнестические сведения, клинические характеристики (вид ургентного осложнения, коморбидность), морфологические характеристики (гистологический тип опухоли, стадия pTNM, характер опухолевого роста (экзо- или эндофитный, смешанный), размер опухоли по длине кишки (до 4 см, 4–7 см, >7 см), наличие метастазов в лим- фатических узлах и отдаленное метастазирование). При помощи многофакторной логистической регрессии про анализированы факторы, оказавшие независимое наиболее статистически значимое влияние на развитие декомпенсированной кишечной непроходимости и перфорации опухоли толстой кишки. Материалы и методы. В ретроспективное когортное исследование включали больных, страдающих колоректальным раком и перенесших экстренные хирургические вмешательства в период с января 2016 г. по январь 2020 г. в об- щехирургических стационарах г. Смоленска. Всего было включено 214 пациентов, медиана возраста пациентов составила 66 лет. Были проанализированы социально-демографические характеристики (пол, возраст, место проживания (городская или сельская местность), образование, трудовая занятость, профессиональный и семейный статусы), анамнестические сведения, клинические характеристики (вид ургентного осложнения, коморбидность), морфологические характеристики (гистологический тип опухоли, стадия pTNM, характер опухолевого роста (экзо- или эндофитный, смешанный), размер опухоли по длине кишки (до 4 см, 4–7 см, >7 см), наличие метастазов в лим- фатических узлах и отдаленное метастазирование). При помощи многофакторной логистической регрессии про анализированы факторы, оказавшие независимое наиболее статистически значимое влияние на развитие декомпенсированной кишечной непроходимости и перфорации опухоли толстой кишки. Результаты. Большее влияние на развитие декомпенсированной кишечной непроходимости оказывали стадия заболевания – местный статус опухоли T4 (отношение шансов (ОШ) 3,19; 95 % доверительный интервал (ДИ) 1,66–7,12; p <0,001), высокая степень злокачественности опухоли G3–G4 (ОШ 2,93; 95 % ДИ 0,89–3,97; p = 0,008) и коморбидность – наличие конкурирующих заболеваний (ОШ 2,03; 95 % ДИ 1,84–2,39; p <0,001). Risk factors for urgent complications of colorectal cancer Risk factors for urgent complications of colorectal cancer S. N. Shchaeva Smolensk State Medical University, Ministry of Health of Russia; 28 Krupskоу St., Smolensk 214019, Russia C o n t a c t s : Svetlana Nikolaevna Shchaeva shaeva30@mail.ru Objective: to identify the main factors associated with an increased risk of urgent complication Materials and methods. This retrospective cohort study included 214 patients with colorectal can emergency surgery in Smolensk hospitals between January 2016 and January 2020. Their media analyzed patients’ sociodemographic characteristics (sex, age, settings (urban or rural), educat fession, and family status), disease history, clinical characteristics (type of urgent complication phological tumor characteristics (histological type, pTNM stage, pattern of tumor growth (exo- S. N. Shchaeva Smolensk State Medical University, Ministry of Health of Russia; 28 Krupskоу St., Smolensk 214019, Rus Smolensk State Medical University, Ministry of Health of Russia; 28 Krupskоу St., Smolensk 214019, Russia Svetlana Nikolaevna Shchaeva shaeva30@mail.ru Svetlana Nikolaevna Shchaeva shaeva30@mail.ru Введение осложнений рассмотрены декомпенсированная острая кишечная непроходимость (ДОКН) и перфорация опухоли, по поводу которых выполнялись хирургичес кие вмешательства по экстренным показаниям в пер- вые 2–4 ч от момента поступления. осложнений рассмотрены декомпенсированная острая кишечная непроходимость (ДОКН) и перфорация опухоли, по поводу которых выполнялись хирургичес кие вмешательства по экстренным показаниям в пер- вые 2–4 ч от момента поступления. Колоректальный рак (КРР) на ранних стадиях в большинстве случаев протекает бессимптомно, од- нако в 15–30 % случаев пациенты с КРР имеют симп томы острого живота, характерные для таких осложне- ний, как перфорация кишечника или острая кишечная непроходимость [1]. Эти осложнения чаще всего воз- никают у лиц пожилого и старческого возраста и зача- стую сопровождаются неблагоприятным прогнозом [2]. При ургентных осложнениях КРР наблюдаются высокая частота послеоперационных осложнений (40–50 %) и высокие показатели периоперационной летальности – до 20 % [3–5]. Критерии включения: больные в возрасте ≥18 лет с осложнениями КРР, потребовавшими экстренных хирургических вмешательств, такими как ДОКН, пер- форация опухоли; гистологический тип опухоли, со- ответствующий аденокарциноме различной степени дифференцировки, перстневидноклеточному и недиф- ференцированному раку. Критерии исключения: пациенты, страдающие первично-множественными злокачественными ново- образованиями, как синхронными, так и метахронными; случаи морфологического типа опухоли, соответст вующего неэпителиальным злокачественным опухо- лям, карциноиду. Острая кишечная непроходимость – наиболее ча- стое осложнение КРР. По данным различных исследо- ваний, частота данного осложнения составляет от 8 до 59 % [6–8]. Непроходимость кишечника чаще встречается при злокачественных опухолях нисходя- щей ободочной кишки из‑за меньшего диаметра левой ободочной кишки. Перфорация опухоли кишечника происходит в результате ишемии и некроза кишечной стенки [9]. Частота перфорации кишечника у пациен- тов с КРР варьирует от 1,2 до 10,0 % [10–12]. Исследование одобрено этическим комитетом ФГБОУ ВО «Смоленский государственный медицин- ский университет» Минздрава России, протокол № 1 от 20.05.2020. В электронную базу данных включены следующие сведения: 1) социально-демографические характери- стики (пол, возраст, место проживания (городская или сельская местность), образование, трудовая за нятость, профессиональный и семейный статусы); 2) анамнестические сведения; 3) клинические харак- теристики: вид ургентного осложнения, коморбидность; 4) морфологические характеристики: гистологический тип опухоли, стадия pTNM, характер опухолевого ро- ста (экзо- или эндофитный, смешанный), размер опу- холи по длине кишки (до 4 см, 4–7 см, >7 см), наличие метастазов в лимфатических узлах и отдаленное мета- стазирование. Цель исследования – изучить основные факторы, оказывающие влияние на развитие ургентных ослож- нений КРР. C o n t a c t s : Objective: to identify the main factors associated with an increased risk of urgent complications of colorectal cancer. Materials and methods. This retrospective cohort study included 214 patients with colorectal cancer who had undergone emergency surgery in Smolensk hospitals between January 2016 and January 2020. Their median age was 66 years. We analyzed patients’ sociodemographic characteristics (sex, age, settings (urban or rural), education, employment, pro- fession, and family status), disease history, clinical characteristics (type of urgent complication, comorbidities), mor- phological tumor characteristics (histological type, pTNM stage, pattern of tumor growth (exo- or endophytic, mixed), 28 Тазовая хирургия и онкология Pelvic Surgery and Oncology The risk of perforation was higher among patients with T4 tumors (OR 3.74; 95 % CI 2.61–5.48; p <0.001), lymph node involvement (N+) (OR 1.61; 95 % CI 1.33–2.01; p <0.001), high-grade tumors G3–G4 (OR 3.56; 95 % CI 2.08–4.93; p <0.001), and diabetes mellitus requiring insulin therapy (OR 2.11; 95 % CI 1.78–2.42; p <0.001). q g py ( p ) Conclusion. Elderly patients with stage III and IV colorectal cancer and high comorbidity are more likely do develop urgent complications, such as decompensated acute intestinal obstruction or tumor perforation. Key words: urgent complications of colorectal cancer, decompensated acute intestinal obstruction, tumor perforation и н а complications of colorectal cancer, decompensated acute intestinal obstruction, tumor perforation For citation: Shchaeva S. N. Risk factors for urgent complications of colorectal cancer. Tazovaya Khirurgiya i Onkologiya = Pelvic Surgery and Oncology 2022;12(2):28–35. (In Russ.). DOI: 10.17650/2686‑9594‑2022‑12‑2‑28‑35. Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 tumor length along the intestine (<4 cm, 4–7 cm, >7 cm), presence of lymph node metastases and distant metastases). Multivariate logistic regression was used to analyze factors that were independently associated with an increased risk of decompensated intestinal obstruction and colon perforation. tumor length along the intestine (<4 cm, 4–7 cm, >7 cm), presence of lymph node metastases and distant metastases). Multivariate logistic regression was used to analyze factors that were independently associated with an increased risk of decompensated intestinal obstruction and colon perforation. Results. The most significant risk factors for decompensated intestinal obstruction included T4 stage (odds ratio (OR) 3 19 95 % fid i t l (CI) 1 66 7 12 0 001) hi h d t G G (OR 2 93 95 % CI 0 89 3 97 tumor length along the intestine (<4 cm, 4–7 cm, >7 cm), presence of lymph node metastases and distant metastases). Multivariate logistic regression was used to analyze factors that were independently associated with an increased risk of decompensated intestinal obstruction and colon perforation. Results. The most significant risk factors for decompensated intestinal obstruction included T4 stage (odds ratio (OR) 3.19; 95 % confidence interval (CI) 1.66–7.12; p <0.001), high-grade tumor G3–G4 (OR 2.93; 95 % CI 0.89–3.97; p = 0.008), and presence of competing diseases (OR 2.03; 95 % CI 1.84–2.39; p <0.001). The risk of perforation was higher among patients with T4 tumors (OR 3.74; 95 % CI 2.61–5.48; p <0.001), lymph node involvement (N+) (OR 1.61; 95 % CI 1.33–2.01; p <0.001), high-grade tumors G3–G4 (OR 3.56; 95 % CI 2.08–4.93; p <0.001), and diabetes mellitus requiring insulin therapy (OR 2.11; 95 % CI 1.78–2.42; p <0.001). О р и г и н а л ь н о е и с с л е д о в а н и е Results. The most significant risk factors for decompensated intestinal obstruction included T4 stage (odds ratio (OR) 3.19; 95 % confidence interval (CI) 1.66–7.12; p <0.001), high-grade tumor G3–G4 (OR 2.93; 95 % CI 0.89–3.97; p = 0.008), and presence of competing diseases (OR 2.03; 95 % CI 1.84–2.39; p <0.001). О р и г и н а л ь н о е и с с л е д о в а н и е Фактор Factor Число пациен- тов, n (%) Number of patients, n (%) Пол: Sex: муж male жен female 113 (52,8) 101 (47,2) Возраст, лет: Age, years: 18–39 40–59 60–69 70–79 80–89 2 (0,9) 43 (20,1) 96 (44,9) 59 (27,6) 14 (6,5) Место проживания: Settings: город urban сельская местность rural 141 (65,9) 73 (34,1) Образование: Education: высшее higher среднее secondary 92 (43,0) 122 (57,0) Профессиональный статус по отраслям: Employed in: промышленность и строительство industry and construction транспорт и связь transport and communications сельское хозяйство agriculture торговля, бизнес trade, business наука, просвещение, культура, здравоохранение science, education, culture, healthcare сфера обслуживания service sector 54 (25,2) 36 (16,8) 23 (10,7) 42 (19,6) 31 (14,5) 28 (13,2) Занятость: Employment: работающий employed неработающий unemployed (pensioner, disabled, etc.) безработный unemployed 99 (46,3) 102 (47,6) 13 (6,1) Семейный статус: Marital status: не замужем / не женат single разведена / разведен divorced замужем / женат married 41 (19,2) 27 (12,6) 146 (68,2) Статистический анализ осуществлен с помощью средств программного обеспечения Statistica 10 и IBM SPSS Statistics 20. Для определения взаимосвязи между предикторными переменными и развитием ургентно- го осложнения применялся множественный логисти- ческий регрессионный анализ. Факторы, показавшие статистическую значимость при однофакторном ана- лизе, в последующем проанализированы в многофак- торном логистическом регрессионном анализе. Уро- вень статистической значимости (p) для включения в многофакторный анализ принят ≤0,05. Тазовая хирургия и онкология Pelvic Surgery and Oncology Тазовая хирургия и онкология Pelvic Surgery and Oncology Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 ТОМ 12 / VOL. 12 2’2022 Таблица 1. Социально-демографические характеристики больных с ургентными осложнениями колоректального рака, n = 214 Table 1. Sociodemographic characteristics of patients with urgent complications of colorectal cancer, n = 214 Таблица 1. Социально-демографические характеристики больных с ургентными осложнениями колоректального рака, n = 214 Table 1. Sociodemographic characteristics of patients with urgent complications of colorectal cancer, n = 214 Таблица 1. Социально-демографические характеристики больных с ургентными осложнениями колоректального рака, n = 214 в соответствии с классификацией TNM 8‑го пере смотра. Также применялась гистологическая градация аденокарциномы (исключая нейроэндокринные но- вообразования) толстой кишки по степени злокачест- венности согласно классификации Всемирной орга- низации здравоохранения (2019). в соответствии с классификацией TNM 8‑го пере смотра. Также применялась гистологическая градация аденокарциномы (исключая нейроэндокринные но- вообразования) толстой кишки по степени злокачест- венности согласно классификации Всемирной орга- низации здравоохранения (2019). Материалы и методы Проведено ретроспективное когортное исследова- ние с участием больных, страдающих КРР и поступив- ших по экстренным показаниям по поводу ургентных осложнений КРР в период с января 2016 г. по январь 2020 г. в общехирургические стационары г. Смоленска, являющиеся клиническими базами кафедр ФГБОУ ВО «Смоленский государственный медицинский универ- ситет» Минздрава России. С целью стадирования онкологического процесса применялась классификация злокачественных опухо- лей TNM (UICC, Международный противораковый союз) 8‑го издания (2017). Стадия заболевания у па циентов, пролеченных до 2017 г., была переоценена Сведения о каждом пациенте собраны в результате анализа историй болезни, протоколов операций, ре анимационных карт, результатов патоморфологичес кого исследования опухоли. В качестве ургентных 29 О р и г и н а л ь н о е и с с л е д о в а н и е При анализе анамнеза заболевания большинство пациентов с ургентными осложнениями КРР, вклю- ченных в данное исследование до развития экстрен- ного осложнения, имели в анамнезе указания на на- личие запоров (70,6 %; 151 из 214), чередование запоров и поносов (30,4 %; 65 из 214). Причем 30,8 % (66 из 214) пациентов обращались по поводу данных жалоб к терапевту или врачу общей практики, тем не менее в силу различных причин необходимого обследования они не получили. Проанализированы факторы, оказавшие наиболее статистически значимое влияние на развитие ДОКН (табл. 3) и перфорации опухоли толстой кишки как осложнения КРР (табл. 4). При помощи многофактор- ной логистической регрессии были определены фак- торы, оказавшие наибольшее влияние на развитие ургентных осложнений. Как следует из таблицы, набольшее влияние на раз- витие ДОКН оказывали стадия заболевания, высокая степень злокачественности опухоли (G3–G4), комор- бидность. Факторами, оказавшими наибольшее влияние на развитие перфорации, были стадия заболевания (местный статус опухоли T4, стадия N+), высокая сте- пень злокачественности (G3–G4), коморбидность – наличие сахарного диабета 2‑го типа, корригируемого инсулинотерапией. Риск развития ургентных осложнений колорек- тального рака – ДОКН, перфорации опухоли – увели- чивается с возрастом, более поздней стадией заболе- вания и у пациентов с выраженной коморбидностью. Результаты После проверки соответствия критериям включе- ния в исследование вошло 214 пациентов. Социально- демографические характеристики больных с ургент- ными осложнениями КРР представлены в табл. 1. Преобладали пациенты старших возрастных групп, медиана возраста больных, включенных в исследова- ние, составила 66 лет. У всех больных, включенных в исследование, диагноз был гистологически подтвер- жден после выполненного экстренного хирурги ческого вмешательства по поводу ДОКН или перфо- рации опухоли. До этапа экстренного оперативного лечения больные к онкологу не направлялись, соот- ветственно, неоадъювантного лечения эти пациенты не получали, КРР был диагностирован интраопера- ционно, морфологическая верификация проведена по результатам послеоперационного гистологическо- го исследования. По месту проживания больные с ургентными осложнениями КРР в данном исследовании распреде- лились следующим образом: городские жители – 65,9 %, жители сельской местности – 34,1 %. Сорок три про- цента пациентов имели высшее образование, 57 % – среднее образование. По профессиональному статусу пациенты распределились относительно равномерно по разным отраслям (см. табл. 1). По изучаемому фак- тору «занятость» работающие составили 46,3 %, нера- ботающие (лица пенсионного возраста) – 47,6 %. Клинико-морфологические характеристики пред- ставлены в табл. 2. Большое число больных с ургент- ными осложнениями КРР имели локализацию опухоли в ободочной кишке (n = 134; 62,6 %), причем злокаче- ственные опухоли сигмовидной кишки с осложнен- ным течением составили 69 случаев, опухоли нисходя- щей ободочной кишки – 17, опухоли поперечной ободочной кишки – 11, опухоли восходящей ободоч- ной – 24, опухоли слепой кишки – 13 случаев. В пря- мой кишке злокачественная опухоль локализовалась 30 Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’2022 2 2022 О р и г и н а л ь н о е и с с л е д о в а н и е Таблица 2. Основные клинико-морфологические характеристики больных с ургентными осложнениями колоректального рака, n = 214 Table 2. Результаты Main clinical and morphological characteristics of patients with urgent complications of colorectal cancer, n = 214 Фактор Factor Число па- циентов, n (%) Number of patients, n (%) Локализация опухоли по МКБ-10: Tumor location (ICD-10): С-18 С-19 С-20 134 (62,6) 14 (6,5) 66 (30,9) Ургентное осложнение колоректального рака: Urgent complication of colorectal cancer: декомпенсированная острая кишечная непроходимость decompensated acute intestinal obstruction перфорация perforation 172 (80,4) 42 (19,6) Коморбидность: Comorbidity: пациенты без сопутствующих заболеваний patients without concomitant diseases заболевания сердечно-сосудистой системы cardiovascular diseases заболевания дыхательной системы respiratory diseases сахарный диабет diabetes mellitus мультиморбидность (присутствие конкурирующих сопутствующих заболеваний) multimorbidity (competing comorbidities) 2 (0,9) 36 (16,8) 27 (12,6) 51 (23,8) 98 (45,9) Стадия злокачественного новообразования по TNM: TNM stage: Т3N0M0 IIA T4aN0M0 IIB T4bN0M0 IIC T3N2aM0 IIIB T4aN1M0 IIIB T3N1M1 IV T4aN1M1 IV T4aN2M1 IV 9 (4,2) 24 (11,2) 26 (12,1) 49 (22,9) 53 (24,8) 17 (7,9) 23 (10,8) 13 (6,1) Гистологическая градация аденокарциномы по степени злокачественности: Histological grade of adenocarcinoma: низкая степень злокачественности (G1–G2) low grade (G1–G2) высокая степень злокачественности (G3–G4) high grade (G3–G4) 43 (20,1) 171 (79,9) Опухолевый рост по длине кишки, см: Tumor length along the intestine, cm: <4 4–7 >7 22 (10,3) 79 (36,9) 113 (52,8) Опухолевый рост по окружности кишки: Tumor growth pattern: экзофитный exophytic эндофитный endophytic смешанный mixed 45 (21,0) 87 (40,7) 82 (38,3) Примечание МКБ 10 Международная классификация Таблица 2. Основные клинико-морфологические характеристики больных с ургентными осложнениями колоректального рака, n = 214 Таблица 2. Основные клинико-морфологические характеристики больных с ургентными осложнениями колоректального рака, n = 214 в верхнеампулярном отделе у 41 пациента, в средне ампулярном – у 18, в нижнеампулярном отделе – у 7 пациентов. Из ургентных осложнений в большем числе наблюдений зафиксирована ДОКН. Опухоли с данным осложнением локализовались как в ободоч- ной, так и в прямой кишке. КРР, осложненный пер- форацией и потребовавший экстренного хирургичес кого вмешательства, локализовался в сигмовидной кишке у 30 пациентов, в ректосигмоидном отделе – у 12, в верхнеампулярном отделе прямой кишки – у 24 пациентов. Большинство пациентов, включенных в данное исследование, имели сопутствующие заболева- ния. По стадиям заболевания у пациентов с ургентными осложнениями КРР преобладала IIIB стадия. Значитель- ное число больных с ургентными осложнениями имели высокую степень злокачественности опухоли (G3–G4) – 79,9 %. В большем числе наблюдений больные с ДОКН имели опухолевый рост по длине кишки 4–7 и >7 см, и эндофитный или смешанный характер опухолевого роста по окружности кишки (табл. 2). Таблица 3. Факторы, влияющие на развитие декомпенсированной острой кишечной непроходимости как осложнения колоректального рака Таблица 3. Факторы, влияющие на развитие декомпенсированной острой кишечной непроходимости как осложнения колоректального рака Таблица 4. Факторы, влияющие на развитие перфорации как ослож- нения колоректального рака Table 4. Factors affecting the risk of perforation as a complication of colorectal cancer Table 3. Factors affecting the risk of decompensated acute intestinal obstruction as a complication of colorectal cancer of colorectal cancer Фактор Factor Отношение шансов (95 % дове- рительный интервал) Odds ratio (95 % confidence interval) p Возраст (≥75 лет) Age (≥75 years) 1,83 (0,92–3,91) 0,01 Место проживания (сельская мест- ность) Settings (rural) 1,32 (1,03–1,49) 0,031 Коморбидность: Comorbidity: cахарный диабет 2‑го типа (пероральные гипогликемические средства) type 2 diabetes mellitus (oral hypoglycemic agents) сахарный диабет 2‑го типа (инсулинотерапия) type 2 diabetes mellitus (insulin therapy) наличие конкурирующих заболеваний competing diseases 1,43 (1,18–2,07) 2,11 (1,78–2,42) 1,97 (1,85–2,53) 0,025 <0,001 <0,001 Местный статус опухоли T4 Local tumor T4 3,74 (2,61–5,48) <0,001 Стадия N+ Stage N+ 1,61 (1,33–2,01) <0,001 Стадия M+ Stage M+ 1,32 (1,09–1,48) 0,013 Высокая степень злокачественности (G3–G4) High-grade tumor (G3–G4) 3,56 (2,08–4,93) <0,001 Опухолевый рост по окружности кишки (эндофитный) Endophytic growth pattern 0,73 (0,61–0,87) <0,001 obstruction as a complication of colorectal cancer Фактор Factor Отношение шансов (95 % доверительный интервал) Odds ratio (95 % confidence interval) p Возраст (≥75 лет) Age (≥75 years) 1,71 (1,19–2,51) 0,002 Место проживания (сельская местность) Settings (rural) 1,34 (1,14–1,47) <0,001 Локализация опухоли (С-18) Tumor location (С-18) 1,27 (1,04–1,39) 0,001 Коморбидность: Comorbidity: заболевания сердечно-сосудистой системы cardiovascular diseases наличие конкурирующих заболеваний competing diseases 1,94 (1,18–2,39) 2,03 (1,84–2,39) <0,001 <0,001 Местный статус опухоли T4 Local tumor T4 3,19 (1,66–7,12) <0,001 Стадия N+ Stage N+ 1,56 (1,28–1,69) 0,023 Стадия M+ Stage M+ 1,27 (1,02–1,38) 0,011 Высокая степень злокачественности (G3–G4) High-grade tumor (G3–G4) 2,93 (0,89–3,97) 0,008 Опухолевый рост по окружности кишки (смешанный) Mixed growth pattern 0,97 (0,65–1,02) 0,04 Опухолевый рост по длине кишки >4 см Tumor length >4 cm along the intestine 1,44 (1,15–1,64) 0,005 в соответствии с рекомендациями онкологов по дости- жении 75‑летнего возраста прекращаются плановые скрининговые обследования на КРР [15]. Однако уро- вень заболеваемости КРР в популяции старше 80 лет составляет около 1 / 3 популяции в возрасте от 50 до 64 лет [16, 17]. Таким образом, отсутствие необхо- димого скринингового обследования наряду с другими социально-демографическими факторами может уве- личивать риск ургентных осложнений КРР у пациен- тов пожилого и старческого возраста, включая и отсут- ствие специфических симптомов, характеризующих клиническую картину КРР [18, 19]. Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 О р и г и н а л ь н о е и с с л е д о в а н и е Таблица 3. Факторы, влияющие на развитие декомпенсированной острой кишечной непроходимости как осложнения колоректального рака Обсуждение В данное исследование включены больные, у ко- торых КРР не был диагностирован до поступления в стационар по экстренным показаниям. 31 О р и г и н а л ь н о е и с с л е д о в а н и е О р и г и н а л ь н о е и с с л е д о в а н и е По результатам данного исследования семейный статус и уровень образования не оказали статистичес ки значимого влияния на развитие ургентных ослож- нений, хотя в некоторых других исследованиях данные факторы оказывали влияние. Другие исследователи объясняли полученные результаты тем, что люди с бо- лее высоким уровнем образования больше осведомлены о здоровом образе жизни и могут иметь более высокий социально-экономический статус. Они также могут пройти своевременное обследование и внеплановые осмотры в частных клиниках, в связи с чем частота ур- гентных осложнений КРР у них меньше [25, 27]. В масштабном общенациональном голландском исследовании, которое включало более 9 тыс. пациен- тов, было подтверждено влияние возраста на развитие ургентных осложнений КРР. Так, в нем у 22 % больных старческого возраста (≥85 лет) КРР впервые проявил- ся в виде ургентных осложнений по сравнению с 12 % в более молодой популяции [22]. Популяционное ис- следование, проведенное в Великобритании, проде- монстрировало, что у пациентов пожилого и старчес кого возраста вероятность возникновения ургентных осложнений КРР в 3 раза выше: в этом исследовании возраст был самым сильным фактором риска возник- новения ургентных осложнений у больных с деменцией и неблагоприятным социально-экономическим фоном [20]. В нашем исследовании, включавшем 214 больных с ургентными осложнениями КРР, возраст оказы- вал существенное влияние на проявление заболевания в виде ургентных осложнений. Так, риск развития ДОКН и перфорации опухоли увеличивался в возра- сте ≥75 лет почти в 2 раза. По данным проведенного исследования, на разви- тие осложнений КРР, требующих экстренной хирур- гической помощи, наибольшее влияние оказала стадия заболевания: при местном статусе опухоли T4 риск развития ДОКН увеличивался в 3 раза, а риск развития перфорации опухоли – в 3,7 раза; наличие стадии N+ увеличивает риск развития ДОКН в 1,5 раза, а риск развития перфорации опухоли – в 1,6 раза. Кроме то- го, по результатам многофакторного анализа, высокая степень злокачественности увеличивает риск перфо- рации в 3,5 раза. По данным других исследований, ургентные осложнения КРР связаны с более высокой частотой местно-распространенного заболевания (Т4) и наличием метастазов в регионарных лимфатических узлах, сосудистой инвазии, отдаленных метастазов [17, 21, 28–30]. Проведенные исследования продемонстрировали, что с возрастом увеличивается частота рака правой половины ободочной кишки [23, 24]. Важно отметить, что в проведенном исследовании существенное влияние на развитие ургентных ослож- нений оказывала высокая степень злокачественности опухоли (G3–G4), увеличивая практически в 3 раза риск развития ДОКН и в 3,6 раза – риск развития перфорации. О р и г и н а л ь н о е и с с л е д о в а н и е Есть несколько исследований о влиянии степени злокачественности на развитие перфорации толстой кишки. Так, B. Bayar и соавт. в своем исследо- вании наблюдали влияние низкой дифференцировки опухоли на риск развития перфорации толстой кишки [31], в другом исследовании S. Ghazi и соавт. [32] при сравнении муцинозных опухолей в 2 группах боль- ных, перенесших плановые хирургические вмешатель- ства и экстренные операции по поводу КРР, отмечали, что в группе экстренных операций у значительного числа больных были опухоли, имеющие в своем соста- ве бо́льшую долю перстневидных клеток. Необходимо отметить, что больные с поздними стадиями, как правило, в большинстве наблюдений по тем или иным причинам обращаются за медицин- ской помощью в более поздние сроки. Это может быть связано с факторами, которые напрямую связаны с па- циентом, а именно с наличием сопутствующей пато- логии со стороны сердечно-сосудистой, дыхательной систем, которая может требовать госпитализации и хи- рургических вмешательств; также наличие старческой деменции может приводить к поздней постановке ди- агноза. Кроме того, такой фактор, как место прожива- ния (сельская местность), влияет таким образом, что даже наличие первых признаков заболевания и своевре- менное обращение пациента за медицинской помощью не всегда гарантируют своевременную диагностику опухолевого процесса, что может приводить к экстрен- ным госпитализациям и установлению диагноза во время операции. В нашем исследовании выявлены различия в развитии ургентных осложнений в зависи- мости от места проживания пациента; так, риск раз- вития ургентных осложнений у больных, проживающих Таблица 3. Факторы, влияющие на развитие декомпенсированной острой кишечной непроходимости как осложнения колоректального рака Также в ряде ис- следований отмечено, что в старческом возрасте на- блюдается более ограниченный доступ к скрининго- вым программам, что оказывает неблагоприятное У пациентов пожилого и старческого возраста, как правило, наблюдается больше сопутствующих па- тологий и может быть больше связанных с ними го- спитализаций, чем у лиц молодого возраста [13]. Они могут находиться на стационарном лечении и прохо- дить диагностические обследования по поводу других заболеваний, и какое‑то время опухолевая патология может быть в ряде случаев вовремя не диагностирован- ной и заболевание впервые может проявиться в виде ургентных осложнений (ДОКН, перфорации опухоли), что значительно увеличивает частоту экстренных го- спитализаций [14]. Несмотря на то, что в развитых странах имеются существенные различия касательно рекомендаций по скринингу КРР, практически во всех странах 32 Тазовая хирургия и онкология Pelvic Surgery and Oncology Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 влияние на своевременную диагностику заболевания и, соответственно, клиническую стадию и прогноз [13–15, 17, 20, 21]. В канадском многоцентровом ис- следовании, включавшем более 41 тыс. пациентов с впервые диагностированным КРР, у 39 % пациентов заболевание впервые проявилось в виде ургентных осложнений, и эти пациенты имели отдаленные мета- стазы, и большинство из этих больных (63 %) были старше 70 лет [21]. По результатам проведенного ис- следования из данных анамнеза всех 214 пациентов установлено, что ранее они не проходили скрининго- вые обследования на КРР. влияние на своевременную диагностику заболевания и, соответственно, клиническую стадию и прогноз [13–15, 17, 20, 21]. В канадском многоцентровом ис- следовании, включавшем более 41 тыс. пациентов с впервые диагностированным КРР, у 39 % пациентов заболевание впервые проявилось в виде ургентных осложнений, и эти пациенты имели отдаленные мета- стазы, и большинство из этих больных (63 %) были старше 70 лет [21]. 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Factors associated with knowledge, attitudes, and practice towards colorectal cancer and its screening among people aged 50–75 years. Int J Environ Res Public Health 2021;18 (8):4100. DOI: 10.3390/ijerph18084100. 27. Thatcher E.J., Camacho F., Anderson R.T. et al. Spatial analysis of colorectal cancer outcomes and socioeconomic factors in Virginia. BMC Public Health 2021;21(1):1908. DOI: 10.1186/s12889-021-11875-6. 18. Renzi C., Lyratzopoulos G., Card T. et al. Do colorectal cancer patients diagnosed as an emergency differ from non- emergency patients in their consultation patterns and symptoms? A longitudinal data-linkage study in England. Br J Cancer 2016;115(7):866–75. DOI: 10.1038/bjc.2016.250. 8. Lee D.B., Shin S., Yang C.S. Patient outcomes and prognostic factors associated with colonic perforation surgery: a retrospective study. J Yeungnam Med Sci 2022;39(2):133–40. DOI: 10.12701/yujm.2021.01445. 28. Neuman H.B., O’Connor E.S., Weiss J. et al. 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DOI: 10.1016/j.amsu.2020.11.001. 23. Qin Q., Yang L., Sun Y.K. et al. Comparison of 627 patients with right- and left-sided colon cancer in China: Differences in clinicopathology, recurrence, and survival. Chronic Dis Transl Med 2017;3(1):51–9. DOI: 10.1016/j.cdtm.2017.02.004. 13. Xing F., Luo R., Chen W., Zhou X. The risk-adjusted Charlson comorbidity index as a new predictor of one-year mortality rate in elderly Chinese patients who underwent hip fracture surgery. Orthop Traumatol Surg Res 2021;107(3):102860. DOI: 10.1016/j.otsr.2021.102860. 3. Wallace D., Walker K., Kuryba A. et al. Identifying patients at risk of emergency admission for colorectal cancer. Br J Cancer 2014;111(3):577–80. DOI: 10.1038/bjc.2014.300. 24. Щаева С.Н. Экстренные резекцион- ные вмешательства у больных ослож- ненным раком правой и левой полови- ны ободочной кишки: отдаленные результаты. Тазовая хирургия и онко- логия 2018;8(2):46–54. [Schaeva S.N. Emergency resections in patients with complicated rightand left-sided colon cancer: long-term outcomes. Tazovaya Khirurgiya i Onkologiya = Pelvic Surgery and Oncology 2018;8(2):46–54. (In Russ.)]. DOI: 10.17650/2220-3478- 2018-8-2-46-54. 4. Schwenter F., Morel P., Gervaz P. Management of obstructive and perforated colorectal cancer. Expert Rev Anticancer Ther 2010;10(10):1613–9. DOI: 10.1586/era.10.147. O : 0. 0 6/j.ots . 0 . 0 860. 14. Askari A., Faiz O. Response to “Emergency presentation and socioeconomic status in colon cancer”. Eur J Surg Oncol 2014; 40(9):1163. Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 ТОМ 12 / VOL. 12 2’2022 О р и г и н а л ь н о е и с с л е д о в а н и е Выводы По результатам данного исследования установле- но, что пациенты старческого возраста с III и IV ста- диями КРР и при наличии высокой коморбидности с большей вероятностью могут иметь ургентные осложнения в виде ДОКН и перфорации опухоли. 33 Тазовая хирургия и онкология Pelvic Surgery and Oncology Л И Т Е Р А Т У Р А / R E F E R E N C E S Perforated tumors in the gastrointestinal tract: CT findings and clinical implications. Br J Radiol 2012;85(1017):1307–13. DOI: 10.1259/bjr/21382039. 29. Askari A., Nachiappan S., Currie A. et al. Who requires emergency surgery for colorectal cancer and can national screening programmes reduce this need? Int J Surg 2017;42:60–8. DOI: 10.1016/j.ijsu.2017.04.050. 20. Wallace D., Walker K., Kuryba A. et al. Identifying patients at risk of emergency admission for colorectal cancer. Br J Cancer 2014;111(3):577–80. DOI: 10.1038/bjc.2014.300. 10. Choo J.M., Baek S.J., Kwak J.M. et al. Clinical characteristics and oncologic outcomes in patients with preoperative clinical T3 and T4 colon cancer who were staged as pathologic T3. Ann Surg Treat Res 2020;99(1):37–43. DOI: 10.4174/astr.2020.99.1.37. 30. Decker K.M., Lambert P., Nugent Z. et al. Time trends in the diagnosis of colorectal cancer with obstruction, perforation, and emergency admission after the introduction of population-based 21. Rabeneck L., Paszat L.F., Li C. Risk factors for obstruction, perforation, or emergency admission at presentation 34 Тазовая хирургия и онкология Pelvic Surgery and Oncology ТОМ 12 / VOL. 12 2’ 2022 organized screening. JAMA Netw Open 2020;3(5):e205741. DOI: 10.1001/jamanetworkopen.2020. 5741. organized screening. JAMA Netw Open 2020;3(5):e205741. DOI: 10.1001/jamanetworkopen.2020. 5741. organized screening. JAMA Netw Open 2020;3(5):e205741. DOI: 10.1001/jamanetworkopen.2020. 5741. 31. Bayar B., Yılmaz K.B., Akıncı M. et al. An evaluation of treatment results of emergency versus elective surgery in colorectal cancer patients. Ulus Cerrahi Derg 2015;32(1):11–7. DOI: 10.5152/UCD.2015.2969. 32. Ghazi S., Berg E., Lindblom A., Lindforss U. Low-Risk Colorectal of emergency versus elective surgery in colorectal cancer patients. Ulus Cerrahi Derg 2015;32(1):11–7. DOI: 10.5152/UCD.2015.2969. 32. Ghazi S., Berg E., Lindblom A., Lindforss U. Low-Risk Colorectal Cancer Study Group. Clinicopathological analysis of colorectal cancer: a comparison between emergency and elective surgical cases. World J Surg Oncol 2013;11:133. DOI: 10.1186/1477-7819-11-133. Cancer Study Group. Clinicopathological analysis of colorectal cancer: a comparison between emergency and elective surgical cases. World J Surg Oncol 2013;11:133. DOI: 10.1186/1477-7819-11-133. Cancer Study Group. Clinicopathological analysis of colorectal cancer: a comparison between emergency and elective surgical cases. World J Surg Oncol 2013;11:133. DOI: 10.1186/1477-7819-11-133. О р и г и н а л ь н о е и с с л е д о в а н и е 31. Bayar B., Yılmaz K.B., Akıncı M. et al. An evaluation of treatment results ORCID автора / ORCID of author С.Н. Щаева / S.N. Shchaeva: https://orcid.org/0000-0002-1832-5255 Конфликт интересов. Автор заявляет об отсутствии конфликта интересов. Conflict of interest. The author declares no conflict of interests. Конфликт интересов. Автор заявляет об отсутствии конфликта интересов. Conflict of interest The author declares no conflict of interests Конфликт интересов. Автор заявляет об отсутствии конфликта интересов. Финансирование. Исследование выполнено без спонсорской поддержки. Financing. The study was performed without external funding. Статья поступила: 30.03.2022. Принята к публикации: 26.04.2022. Article submitted: 30.03.2022. Accepted for publication: 26.04.2022. Соблюдение прав пациентов и правил биоэтики. Протокол исследования одобрен ком Соблюдение прав пациентов и правил биоэтики. Протокол исследования одобрен комитетом по биомедицинской этике ФГБОУ ВО «Смо- ленский государственный медицинский университет» Минздрава России (протокол № 1 от 20.05.2020). Все пациенты подписали инфор- мированное согласие на участие в исследовании Соблюдение прав пациентов и правил биоэтики. Протокол исследования одобрен комитетом по биомедицинск р у д Compliance with patient rights and principles of bioethics. The study protocol was approved by the biomedical ethics committee of Smolensk State Medical University, Ministry of Health of Russia (protocol No. 1 dated 20.05.2020). All patients signed written informed consent to participate in the study. Статья поступила: 30.03.2022. Принята к публикации: 26.04.2022. Article submitted: 30.03.2022. Accepted for publication: 26.04.2022. Статья поступила: 30.03.2022. Принята к публикации: 26.04.2022. Article submitted: 30.03.2022. Accepted for publication: 26.04.2022. 35
https://openalex.org/W2468858795	https://europepmc.org/articles/pmc4759530?pdf=render	English	null	The Influence of Structure Heights and Opening Angles of Micro- and Nanocones on the Macroscopic Surface Wetting Properties	Scientific reports	2,016	cc-by	7,238	Ling Schneider, Milan Laustsen, Nikolaj Mandsberg & Rafael Taboryski We discuss the influence of surface structure, namely the height and opening angles of nano- and microcones on the surface wettability. We show experimental evidence that the opening angle of the cones is the critical parameter on sample superhydrophobicity, namely static contact angles and roll- off angles. The textured surfaces are fabricated on silicon wafers by using a simple one-step method of reactive ion etching at different processing time and gas flow rates. By using hydrophobic coating or hydrophilic surface treatment, we are able to switch the surface wettability from superhydrophilic to superhydrophobic without altering surface structures. In addition, we show examples of polymer replicas (polypropylene and poly(methyl methacrylate) with different wettability, fabricated by injection moulding using templates of the silicon cone-structures. Tuning surface wettability by engineering surface structure and/or surface chemistry has attracted huge research interests, due to the promising applications of these surfaces in areas of self-cleaning, anti-icing, drag reduction, anti-fogging and microfluidic devices1–12. A general approach is to texture low surface energy materials to obtain superhydrophobic surfaces, and high surface energy materials to obtain superhydrophilic surfaces. These artificial structures are typically inspired by nature, for example, by lotus leaves13, water strider’s legs14, moth eyes3 and etc15,16. In addition to creating biomimetic structures, there are also studies, where periodic artificial structures of different geometry are fabricated for obtaining a better theoretical understanding of the wetting phenomena to benefit the future structural design17–21. Though there have been a few studies on the influence of height, width, and period of nano- and micro-cones on the surface wettability9,22, the influence of the structural include/open- ing angles on the surface wettability has rarely been discussed. The fabrication of tapered cones require some predesigned templates2,23,24, pre-moulding methods25, or fine adjustment of processing parameters26,27, which makes it difficult to tune the opening angles of the cones. However, the cone geometry is usually closer to the aforementioned natural structures than the flat pillars or holes; and it has been suggested that the opening angle of the cones can have crucial influence on the sample wettability9,28. l Reactive ion etching (RIE) is a dry etching technique that can be used to structure silicon surfaces without masks through the combined effect of a corrosive gas (SF6 and/or CH4) and a passivating gas (O2)26,29–32. www.nature.com/scientificreports www.nature.com/scientificreports www.nature.com/scientificreports Department of Micro- and Nanotechnology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. Correspondence and requests for materials should be addressed to L.S. (email: lisu@nanotech.dtu.dk) or R.T. (email: rata@nanotech.dtu.dk) The Influence of Structure Heights and Opening Angles of Micro- and Nanocones on the Macroscopic Surface Wetting Properties received: 16 September 2015 accepted: 22 January 2016 Published: 19 February 2016 received: 16 September 2015 accepted: 22 January 2016 Published: 19 February 2016 Ling Schneider, Milan Laustsen, Nikolaj Mandsberg & Rafael Taboryski Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 received: 16 September 2015 accepted: 22 January 2016 Published: 19 February 2016 Ling Schneider, Milan Laustsen, Nikolaj Mandsberg & Rafael Taboryski (a) Representative scanning electron microscopy images (20° tilted view and cross-sectional view) of nano- and submicron cones prepared with different flow rates of SF6 and O2 gases. The flow rates of SF6 and O2 vary from 70–50, 70–70, 70–90, to 70–130 sccm. For simplicity, we denote samples of different SF6 and O2 flow rates and etching time as − − Q Q t SF O 6 2 . Therefore 70–50–8 means a sample processed with QSF6 = 70 sccm, and QO2 = 50 sccm for 8 min. The detailed processing information can be found in the supporting information (SI). Scale bars are 1 μ m. (b) Images of sessile water droplets on flat and structured Si surfaces with FDTS coating, and the corresponding measured static contact angles. Droplet volume is 10 μ l. Figure 1. (a) Representative scanning electron microscopy images (20° tilted view and cross-sectional view) of nano- and submicron cones prepared with different flow rates of SF6 and O2 gases. The flow rates of SF6 and O2 vary from 70–50, 70–70, 70–90, to 70–130 sccm. For simplicity, we denote samples of different SF6 and O2 flow rates and etching time as − − Q Q t SF O 6 2 . Therefore 70–50–8 means a sample processed with QSF6 = 70 sccm, and QO2 = 50 sccm for 8 min. The detailed processing information can be found in the supporting information (SI). Scale bars are 1 μ m. (b) Images of sessile water droplets on flat and structured Si surfaces with FDTS coating, and the corresponding measured static contact angles. Droplet volume is 10 μ l. we will show some examples of polymer surfaces (polypropylene, PP and poly(methyl methacrylate), PMMA) with different wettability. Our technique will pave the way for the design and fabrication of large scale and low cost functional surfaces. Ling Schneider, Milan Laustsen, Nikolaj Mandsberg & Rafael Taboryski As most of the surfaces made this way appear black due to the scattering of the incident light by nano- and submicron structures, this method is also known as the “black silicon method”, and has widely been used in optical appli- cations such as anti-reflective surfaces26,31,32. By tuning the processing parameters, namely gas flow rate, platen power, chamber pressure, and etching time, it is possible to alter the structure shape, and even the opening angles of the cones9. However, to the best of our knowledge, there are not yet reports on applying this method to system- atically study the influence of structural opening angle on the surface wettability. y yl g g y In this paper, we report a systematic study of surface wettability versus structural opening angles of the cones. We fabricate the nano- and microcones (structure heights range from less than 100 nm to around 2 μ m) using RIE and alter the structural geometry by varying the processing time and gas flow rates. By using hydrophobic coating or hydrophilic surface treatment, we are able to switch the surface wettability from superhydrophilic to superhydrophobic without altering surface structures. We will discuss the influence of structure height and open- ing angle on sample superhydrophobicity, measured as the static contact angle and the roll-off angle. In addition, Department of Micro- and Nanotechnology, Technical University of Denmark, 2800 Kongens Lyngby, Denmark. Correspondence and requests for materials should be addressed to L.S. (email: lisu@nanotech.dtu.dk) or R.T. (email: rata@nanotech.dtu.dk) 1 Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 www.nature.com/scientificreports/ Figure 1. (a) Representative scanning electron microscopy images (20° tilted view and cross-sectional view) of nano- and submicron cones prepared with different flow rates of SF6 and O2 gases. The flow rates of SF6 and O2 vary from 70–50, 70–70, 70–90, to 70–130 sccm. For simplicity, we denote samples of different SF6 and O2 flow rates and etching time as − − Q Q t SF O 6 2 . Therefore 70–50–8 means a sample processed with QSF6 = 70 sccm, and QO2 = 50 sccm for 8 min. The detailed processing information can be found in the supporting information (SI). Scale bars are 1 μ m. (b) Images of sessile water droplets on flat and structured Si surfaces with FDTS coating, and the corresponding measured static contact angles. Droplet volume is 10 μ l. Figure 1. Results and Discussionl (a) Side view of a partially wetted cone; (b) column graph of samples of different opening angles fabricated by different processing parameters ( , Q Q SF O 6 2 and t). The detailed processing parameters, and the corresponding heights, opening angles, and contact angles can be found in Table S1. that most samples exhibit a (tapered) cone shape, the geometry of these samples is determined by only two parameters, the height and the opening angle α 2 of the cones (Fig. 2a). It is thus necessary to discuss which geometric parameters dominate the wetting properties of a chemically fixed surface.l i As depicted in Fig. 2a, the water-air interface is almost flat on the microscopic scale at a quasi-equilibrium9,28. The micro-contact angle at the water-solid sidewall θµ (Fig. 2a) is thus determined by the advancing contact angle θA of the flat FDTS surface. This is to say, for the non-wetting condition, the Cassie-Baxter state35, or an interme- diate Cassie-impregnating state28,36, θ α θ = + ° ≤ µ 90 A; while for the complete wetting situation, the Wenzel state37, θ α θ > + ° > µ 90 A. Hence for the non-wetting condition, the opening angle of an individual cone has to fulfil 2α ≤ 51.0° ± 1.8°. Figure 2b summarizes the opening angles of 14 samples and their corresponding wetting states. Among all 14 measured samples, two are sticky, that is water droplets were pinned to the surface even when the surface was positioned upside down; and the rest are non-sticky, that is water droplets rolled/slid off from the surfaces at a certain surface inclination. Apparently, all the non-sticky samples have opening angles smaller than the threshold value 51.0° ± 1.8°. The two sticky samples have however opening angles of α= 2 49.5° ± 10.2° (70– 130–8) and 2α = 47.8° ± 7.4° (70–130–12), which are above or comparable to the expected threshold. Interestingly, for Si-70–130–8, all 5 tested drops were pinned; while for Si-70–130–12, only 3 out of 5 drops were pinned, which fits very well with this threshold hypothesis. It is worth mentioning that here we use the critical angle to draw a distinct line between the slippery and sticky surfaces. However, the degree of slipperiness and stickiness is always gradual. Results and Discussionl In this study, we altered the flow rate of SF6 and O2 and the etching time t, and kept the rest of the parameters unchanged, as the gas flow rate is the governing parameter for tuning the opening angles of the resulting cones26,31,32. As the absorbance of light varies with the geometric features of the structures, such as, height, aspect ratio, and period of the structures26,31–34, our samples do not necessarily appear black; some are black and some appear brownish. Superhydrophobic silicon samples. Figure 1a shows scanning electron microscopy (SEM) images of Si surfaces prepared by RIE for 8 min, using SF6 of 70 sccm and O2 of various flow rates: 50, 70, 90, and 130 sccm respectively. For simplicity, we denote samples of different SF6 and O2 flow rates and etching time as − − Q Q t SF O 6 2 . Therefore 70–50–8 means a sample processed with QSF6 = 70 sccm, and QO2 = 50 sccm for 8 min. Morphologies of samples vary from sharp hierarchical needles (70–50–8), tapered cones (70–70–8 and 70–90–8), to rounded domains (70–130–8), which are macroscopically uniform while microscopically stochastic. According to the cross-sectional SEM images, structures prepared with different O2 flow rates are of different heights: with increasing O2 rate, the structure height reduces from 1.540 ± 0.544, 0.733 ± 0.079, 0.557 ± 0.084, to 0.264 ± 0.026 μ m. This trend is consistent with results obtained by other groups26,31,32. As the height of the fabri- cated structures ranges from a few hundred nanometers to a few micrometres, we refer to our structures as nano- and microcones. Results of static water contact angle measurements of these surfaces in comparison to the flat Si surface are shown in Fig. 1b. For samples with the hydrophobic 1 H, 1 H, 2 H, 2 H-perflourodecyltrichlorosilane (FDTS) coating, with increasing O2 flow rate, we measured decreasing static water contact angle. By assuming Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 2 www.nature.com/scientificreports/ Figure 2. (a) Side view of a partially wetted cone; (b) column graph of samples of different opening angles fabricated by different processing parameters ( , Q Q SF O 6 2 and t). The detailed processing parameters, and the corresponding heights, opening angles, and contact angles can be found in Table S1. Figure 2. Results and Discussionl In fact, the wetting test on sample Si-70–130–12 shows that depending on structural imperfections on the microscale, the surface can be either “slippery” and “sticky” on macroscale. Th h l f b fi h l l h d h fl f h Though results from Fig. 2b fit the critical opening angle assumption without considering the influence of the structural height, the influence of structural height on the wetting properties of individual samples should not be excluded completely. To understand the influence of the opening angles and structure heights, we plot water contact angle and roll-off angle of different samples with respect to both H and α 2 (Fig. 3). Interestingly, the static and dynamic contact angles do not show obvious dependence on H but exhibit an apparent linear dependence on 2α for the cones.h The apparent contact angle at equilibrium θe of a rough surface is usually described by the Cassie-Baxter equa- tion for the non-wetting state9,35 θ θ = ( + ) − , ( ) f cos cos 1 1 1 e s Y ( ) 1 where = + f s A A A ls ls lg, is the solid fraction. ls lg For a cone geometry depicted in Fig. 2a, π α α = . ( ) A h tan cos 2 ls 2 ( ) 2 α π α = − ≥ A H h 2 3 tan tan 0 lg 2 2 2 2 , for hexagonal close-packed cones. Equation 1 can be rewritten a α π α = − ≥ A H h 2 3 tan tan 0 lg 2 2 2 2 , for hexagonal close-packed cones. Equation 1 can be rewritten as, ( ) ( ) ( ) θ π θ π α π = ( + )    −    + − . ( ) cos cos 1 2 3 sin 1 3 e h H Y h H h H 2 2 2 ( ) 3 According to the Cassie-Baxter equation (Eq. 1), the larger the solid fraction f s, the smaller the θe, as both θe and the Young contact angle θY are larger than 90°. The θe ~ 2α dependence from Fig. 3b can only be achieved if h/H increases with increasing 2α, indicating that the penetration depth h depends on 2α for the given geometry. According to the Cassie-Baxter equation (Eq. Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 Results and Discussionl 1), the larger the solid fraction f s, the smaller the θe, as both θe and the Young contact angle θY are larger than 90°. The θe ~ 2α dependence from Fig. 3b can only be achieved if h/H increases with increasing 2α, indicating that the penetration depth h depends on 2α for the given geometry. Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 3 www.nature.com/scientificreports/ Figure 3. Dependence of static contact angles (black squares) and roll-off angles (red circles) on H (a) and α 2 (b) of nano- and submicron cones; (c) dependence of water penetration degree on opening angle, h/H vs sin α ; (d) dependence of the roll-off angle on the contact angle hysteresis, θ − sin Roll off vs θ θ ( − ) cos cos R A . The dash lines serve as guides to the eye, while the solid lines are linear (b,c) and polynomial (d) fits of the experimental data. Figure 3. Dependence of static contact angles (black squares) and roll-off angles (red circles) on H (a) and α 2 (b) of nano- and submicron cones; (c) dependence of water penetration degree on opening angle, h/H vs sin α ; (d) dependence of the roll-off angle on the contact angle hysteresis, θ − sin Roll off vs θ θ ( − ) cos cos R A . The dash lines serve as guides to the eye, while the solid lines are linear (b,c) and polynomial (d) fits of the experimental data. Such dependence is consistent with our data analysis, as shown in Fig. 3c, where the water penetration degree h/H is shown to depend on 2α linearly. Here h/H is calculated by Equation 3, using the experimentally measured angles 2α and θe, and ranges from 5% up to 50%.h g e g p The wetting situations described by the Cassie and Wenzel models are usually considered binary; there is no gradual transition from Cassie to Wenzel state (or vice versa); a surface should be either non-wetted or completely wetted. However, several studies have revealed that an intermediate wetting state exists, which can be reversed back to the Cassie state when certain conditions are met16,28,36,38. Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 Results and Discussionl Figure 4. (a) schematic steps of fabrication of polymer replicas; (b) 2D and 3D AFM images of PP replicas fabricated via two different silicon masters 70–70–8 (H ~ 120 nm), and 70–90–8 (H ~ 460 nm); (c) Images of water droplets on flat and structured PP surfaces. length λ along the rolling direction should be used18,21. For the hexagonal closed packed cone model geometry, this length is given by ( ) λ = ⋅ + α α − d 1 h H 1 sin sin . The Furmidge equation can thus be rewritten as: length λ along the rolling direction should be used18,21. For the hexagonal closed packed cone model geometry, this length is given by ( ) λ = ⋅ + α α − d 1 h H 1 sin sin . The Furmidge equation can thus be rewritten as: θ γ ρ α α θ θ =    − +   ⋅( − ). ( ) − d gV h H sin 1 sin sin 1 cos cos 5 Roll off R A ( ) 5 When we plot the roll-off angle versus contact angle hysteresis (Fig. 3d), we are able to fit the plot with a second order polynomial fit. The second order dependence implies, for our structures, that λ has a linear dependence on the contact angle hysteresis. Such dependence is supported by the plot of λ versus contact angle hysteresis (Figure S3). It is worth mentioning that Equations 3 and 5 are derived based on the hexagonal closed packed cone geom- etry, which is the most representative geometry of the fabricated nano and microcones. However, not all samples exhibit the closed packed geometry, as can be seen from both Fig. 1 and Fig. S1. In the supporting information (SI), we discuss several other packing geometries, which nonetheless only change some prefactors in Equations 3 and 5. Superhydrophobic polymers. Though much effort has been invested to study the wetting properties of silicon samples, the final goal is to develop a technique that can be integrated into existing fabrication techniques for solid materials, such as injection moulding of polymers. As a starting point and a simple demonstration of our technique, we chose structures 70–70–8 and 70–90–8 for master origination and a low surface energy material polypropylene (PP, γ PP ≈ 29 mN/m40) for injection moulding. Results and Discussionl Although we here use the Cassie-Baxter model to explain the linear dependence of θ α ~ cos sin e , it does not necessarily mean that the intermediate state does not exist for some of the geometries presented.hf We now turn to describe the dynamic properties of the surfaces. The linear increase of roll-off angle with increasing opening angle shown in Fig. 3b also indicates that the structures are partially wetted with various water penetration degrees. Such a wetting situation can be understood in two ways: 1. The structures are not perfect – some are sharper and others are blunter. Therefore, there is a stochastic mixture of totally non-wetted and wetted cones on the microscale, which results in the partial wetting situation on macroscale. 2. In an ideal situation, all cones are identical and arranged in hexagonal close packed order. The cones are wetted exactly the same way on the microscale with certain degree of water penetration. The wetting situation of structures fabricated by RIE mostly falls into the first category due to the nature of the RIE technique29. The macroscopic wetting behaviour is none-the-less the same for both cases.hf The roll-off angle θ − Roll off is determined by the balance of the driving force along the rolling plane and the friction during the movement, which is also called the pinning force, and is proportional to the contact angle hysteresis and the surface tension of the liquid. For a flat surface, the relation between a roll-off angle and the contact angle hysteresis can be explained by the Furmidge equation39. θ γ ρ θ θ = ⋅( − ), ( ) − d gV sin cos cos 4 Roll off R A ( ) 4 where ρgV is the gravity force acting on the droplet, which is constant for all measured droplets, d is the width of the drop base viewed along the rolling/sliding direction, and θR and θA are the receding and advancing contact angles respectively. For rough surfaces, instead of using the width d of the drop, the effective three-phase contact Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 4 www.nature.com/scientificreports/ Figure 4. (a) schematic steps of fabrication of polymer replicas; (b) 2D and 3D AFM images of PP replicas fabricated via two different silicon masters 70–70–8 (H ~ 120 nm), and 70–90–8 (H ~ 460 nm); (c) Images of water droplets on flat and structured PP surfaces. Superhydrophilic silicon samples. The complete wetting can be described by the Wenzel equation37 Superhydrophilic silicon samples. The complete wetting can be described by the Wenzel equation37 θ θ = ⋅ , ( ) r cos cos 6 e f Y ( ) 6 θ θ = ⋅ , r cos cos e f Y with = + π α r 1 f 2 3 sin being the surface roughness factor for the cone shape model, which equals the surface area divided by the apparent area of the rough surface. For θY < 90°, creating surface roughness would lead to more hydrophilic surfaces, and even superhydrophilic surfaces, according to the Wenzel model. The water contact angle of unstructured Si surface after plasma cleaning is 35.5 ± 4.1°, and > r 2 f for all samples, as calculated from the opening angle 2α measured on the SEM images. The resulting θe should thus be much smaller than 10° for the structured surfaces. We therefore tested the wetting properties of samples processed by the same SF6 and O2 flow rates as the ones shown in Fig. 1a, but without FDTS coating (Fig. 5). All surfaces exhibited strong superhydro- philicity during the measurements: the water droplet spread on the surface immediately after impacting the sur- face. The static contact angles of all samples are smaller than 5 degree. Interestingly, according to the Wenzel equation, for complete wetting, the higher the surface roughness, the smaller is the contact angle if θY < 90°. However, here we find the highest contact angle ∼ 10° for the surface with the highest r f . The high water contact angle of 70–50–8 is perhaps the result of the hierarchical nature of this sample surface (Fig. 1a). Hejazi et al. sug- gested that for hierarchical surfaces there could be two wetting states at different levels of roughness; structures that contribute to the first order roughness, the small spikes on top of the cones are completely wetted, while structures that contribute to the second order roughness, the cones might be partially wetted (air trapped)45. Such a multi-degree wetting situation can result in a higher apparent contact angle θe, as demonstrated by Hejazi et al., which is consistent with our experimental observation on superhydrophilic Si-70–50–8. Hydrophilic polymer replicas. For a simple demonstration of our method, we textured PMMA to generate the hydrophilic function, as the surface energy of PMMA (γPMMA ≈ 39 mN/m40) is relatively high among polymers and can be injection-moulded. Results and Discussionl Though Si-70–50–5 exhibits the strongest superhy- drophobicity, replication fidelity of this structure could be low due to its hierarchical nature; while 70–70–8 and 70–90–8 with FDTS coating still have high superhydrophobicity and are easy to replicate. The main steps for rep- licating polymer surfaces through Si masters are shown in Fig. 4a. The prepared Si master serves as a template for forming textured Ni shim, and is removed by warm KOH bath after the Ni formation (supporting information). The Ni shim is then used as a mould for injection-moulding textured PP samples. A more detailed fabrication procedure can be found in the supporting information (SI) and our previous publications28,41.t p pp g p p Both replicated PP samples exhibit cone morphology after injection moulding (Fig. 4b), indicating that the structures have been transferred from their Si masters successfully. However, the cones of PP-70–70–8 are much shorter than those of PP-70–90–8, though their masters are quite the opposite: the cones of Si-70–70–8 are ~170 nm higher than those of Si-70–90–8. The replication quality of injection moulded polymer samples depends mainly on the filling quality of the polymer melt and the demoulding of the structures from the mould. The filling of micro-cavities in Ni-70–70–8 is more difficult than in Ni-70–90–8, as the former has a higher aspect ratio (Si- 70–70–8 around 3.6 and Si-70–90–8 around 2.1)42,43. On the other hand, the higher surface area of structure 70–70–8 results in a higher friction during the demoulding, which might cause deformation or even fracture of the moulded polymer cones44. Interestingly, the water wettability of the two samples however follows the trend of their silicon masters: PP-70–70–8 is more superhydrophobic than PP-70–90–8, as summarized in Table 1. As discussed previously, the structure height contribute less to the superhydrophobicity compared to the structure Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 5 www.nature.com/scientificreports/ Si 70–70–8 PP 70–70–8 Si 70–90–8 PP 70–90–8 Static contact angle (°) 161.4 ± 1.4 157.7 ± 2.8 150.2 ± 0.9 150.8 ± 1.6 f s (%) 8.7 8.2 22.1 13.9 roll–off angle (°) 5.6 ± 2.2 13.5 ± 1.6 27.5 ± 0.9 24.8 ± 2.4 Table 1. Static contact angles and roll–off angles of PP replicas and their corresponding Si masters. fs was calculated from the Cassie equation (Eq. 3). Table 1. Static contact angles and roll–off angles of PP replicas and their corresponding Si masters. Results and Discussionl fs was calculated from the Cassie equation (Eq. 3). Figure 5. Images of droplets on uncoated silicon surfaces processed at different etching parameters: 70–50–8, 70–70–8, 70–90–8, and 70–130–8. The photos were taken while the droplet was still impacting the surface, so the water contact angle measured here should be the advancing contact angle. Table 1. Static contact angles and roll–off angles of PP replicas and their corresponding Si masters. fs was calculated from the Cassie equation (Eq. 3). Figure 5. Images of droplets on uncoated silicon surfaces processed at different etching parameters: 70–50–8, 70–70–8, 70–90–8, and 70–130–8. The photos were taken while the droplet was still impacting th surface, so the water contact angle measured here should be the advancing contact angle. pening angle or the structure aspect ratio, which explains why samples with shorter cones exhibit higher uperhydrophobicity. l f h f h l h h d h b l d d b h f For samples of the same surface morphology, their superhydrophobicity is mainly dominated by the surface chemistry. As FDTS has a lower surface energy than PP, the resulting Si surface coated with FDTS should be more superhydrophobic than the PP surface, if we assume both have the same surface structure. The measured water contact angles of Si-70–70–8 and PP-70–70–8 follow such a rule, i.e. Si-70–70–8 is slightly more superhydropho- bic than PP-70–70–8. However, PP-70–90–8 is more superhydrophobic than its silicon master Si-70–90–8. Such a deviation from the theory could possibly be due to the fact that the structure of PP-70–90–8 is different than its Si master Si-70–90–8. During demoulding, the PP cones might be stretched, which might form sharper cones than their Si masters. Methods Fabrication of superhydrophobic and superhydrophilic silicon samples. Silicon wafers (100) with nano- and microcones were fabricated using reactive ion etching (RIE, Pegasus D-RIE, STS, UK). By tuning SF6 and O2 flow rate and etching time during the etching, nano- and microcones of different heights and aspect ratios were fabricated. Detailed fabrication parameters can be found in the supporting information (SI). After RIE, all samples were cleaned by N2/O2 plasma (N2 400 sccm, O2 70 sccm, power 1000 W) for 30 min. For superhydro- phobic applications, an additional monolayer of 1 H, 1 H, 2 H, 2 H-perflourodecyltrichlorosilane (FDTS) was deposited by molecular vapor deposition (MVD, MVD 100, Applied Microstructures, USA) immediately after the plasma treatment, using a standard recipe. The detailed parameters can be found in the supporting information (SI). While for superhydrophilic applications, the samples were used directly after the plasma treatment. Fabrication of nickel mold. Nickel vanadium (NiV) of 100 nm was sputter-coated (CMS-18, Kurt J. Lesker Company® , USA) onto the Si master wafers to serve as a seed layer. A nickel (Ni) layer of 300 μ m was deposited via electroforming. The Si template was then dissolved in a warm KOH bath (30 wt%, 80 °C). A monolayer of 1 H, 1 H, 2 H, 2 H-perflourodecyltrichlorosilane (FDTS) was coated by molecular vapor deposition (MVD, MVD 100, Applied Microstructures, USA) onto the resulting Ni shim. This layer was used as an anti-sticking layer for removing the polymer parts from the mold. Fabrications of both Si masters and Ni molds were done in the clean- room (class 10–100). Injection molding of polymer samples. Polymer samples were injection molded using a variotherm heating system, where the mold temperature is first heated to a higher temperature for maximal filling and cooled down to a lower temperature for easy demolding. Contact angle measurements. Contact angles were measured using an optical tensiometer (Theta, Attension, Finland) with a high-speed camera (3000 fps, MotionXtra N3, IDT, USA) and tilted cradle. Shapes of droplets were fitted by polynomial fitting for both static and dynamic contact angle measurements. For all contact angle measurements presented in the main text, droplets of 10 μ L in volume were used. To ensure that the meas- ured static contact angles do not depend on the drop volume, we compared the measurements on same samples obtained with droplets of both 5 and 10 μ l in volume (Figure S2). Superhydrophilic silicon samples. The complete wetting can be described by the Wenzel equation37 To further improve the wettability of the textured surface, we treated the textured surface with O2 plasma for 25 s. As proved by AFM results (Figure S6), there is no obvious surface structure mod- ification within such a short treatment. i Figure 6 represents PMMA samples fabricated through the silicon master 70–70–8, which is the same one used for fabricating PP samples. Here the height of the cones is only around 120 nm, which is similar to that of PP–70–70–8, but also much smaller than that of the silicon master Si–70-70–8. Such a small height is probably due to the same reason: the polymer melt did not fully fill the Ni cavities, and/or the cones broke during the demolding. The Ni insert used for injection moulding was examined by SEM afterwards, however no obvious residual polymer was found in the holes (Figure S7). We therefore attribute the low structure height to the incom- plete filling of the submicron cavities. Though the structures are a bit lower than expected, the surface wettability has increased after texturing: The water contact angle is θ ≈ ° 67 PMMA for flat untreated PMMA, θ ≈ ° − 57 PMMA t 6 Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 www.nature.com/scientificreports/ www.nature.com/scientificreports/ Figure 6. 2D (a) and 3D (b) AFM images of treated PMMA replica fabricated via silicon masters 70–70–8 (H~120 nm); (c) Photos of water droplets on flat (with and without plasma treatment) and textured PMMA (plasma treated) surfaces. Figure 6. 2D (a) and 3D (b) AFM images of treated PMMA replica fabricated via silicon masters 70–70–8 (H~120 nm); (c) Photos of water droplets on flat (with and without plasma treatment) and textured PMMA (plasma treated) surfaces. for treated PMMA and θ ≈ ° − − − 27 PMMA 70 70 8 for treated and textured PMMA. We can thus conclude that by texturing and surface treating PMMA, we can mass–produce surfaces with high water wettability.l In summary, we have discussed the influencing geometric parameters on surface wettability at a given surface chemistry. Superhydrophilic silicon samples. The complete wetting can be described by the Wenzel equation37 From our experimental results, the opening angles of the nano- and microcones play a dominant role on both the static contact angle and the roll-off angle of a surface: the static and dynamic contact angles of supe- rhydrophobic silicon samples exhibit linear dependence on the opening angles of the cones, whereas there is no obvious dependence of them on the height of the cones. Such a rule also applies to polymer samples, injection moulded using the corresponding silicon masters. In addition, we have demonstrated that the same structures used for superhydrophobic applications also can be used to fabricate superhydrophilic surfaces, without any fur- ther surface modifications required. The multi-order roughness of the hierarchical surface, however, reduces the superhydrophilicity slightly, due to the different wetting states of the various levels of roughness. Methods The deviations of the static contact angles are within the error range. The baseline was determined manually. For each sample, five measurements were made at different locations on the surface. Characterization of nano- and microcones. Structured Si samples were characterized by scanning elec- tron microscopy (SEM, Supra 40 VP, Carl Zeiss AG, Germany) both at cross-sectional view and at 20° tilted view. The heights of the nano- and microcones were determined using software provided by Zeiss and the opening angles were manually measured by Image J. 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Injection molding of high aspect ratio sub-100nm nanostructures. J Micromech Microeng 23, 025003 (2013).f 5. Hejazi, V., Moghadam, A. D., Rohatgi, P. & Nosonovsky, M. Beyond Wenzel and Cassie–Baxter: Second-Order Effects on the Wetting of Rough Surfaces. Langmuir 30, 9423–9429 (2014). 8 Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 www.nature.com/scientificreports/ Author Contributions L.S. designed the project, conducted the experiments, and wrote the manuscript. M.L. fabricated the Ni shim and polymer samples. N.M. conducted the experiments related to hydrophilic PMMA samples. R.T. contributed to the discussion and reviewed the manuscript. Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 Additional Information upplementary information accompanies this paper at http://www.nature.com/srepihi Supplementary information accompanies this paper at http://www.nature.com/srepihi Competing financial interests: The authors declare no competing financial interests. Competing financial interests: The authors declare no competing financial interests. How to cite this article: Schneider, L. et al. The Influence of Structure Heights and Opening Angles of Micro- and Nanocones on the Macroscopic Surface Wetting Properties. Sci. Rep. 6, 21400; doi: 10.1038/srep21400 (2016). This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ Scientific Reports \| 6:21400 \| DOI: 10.1038/srep21400 9
https://openalex.org/W4200278952	https://www.researchsquare.com/article/rs-1184837/latest.pdf	English	null	Evolution of Physicochemical Properties of 2-(2-(4-(4-chloro) phenyl) vinyl)-1, 1, 3-trimethyl-1H-benzo[e] Indolium Iodide via Experimental and Quantum Chemical Calculation for Third-Harmonic Generation Applications	Research Square (Research Square)	2,021	cc-by	5,577	Evolution of Physicochemical Properties of 2-(2-(4- (4-chloro) phenyl) vinyl)-1, 1, 3-trimethyl-1H-benzo[e] Indolium Iodide via Experimental and Quantum Chemical Calculation for Third-Harmonic Generation Applications Eniya Palaniyasan Periyar University Anbarasan Radhakrishnan Periyar University Shivaraj R. Maidur HKBK College of Engineering Parutagouda Shankaragouda Patil K.L.E. Institute of Technology Kalyana Sundar Jeyaperumal (  jksundar50@gmail.com ) Periyar University Abstract In this work, the authors have reported the synthesis, crystal structure, and physicochemical properties of new Indolium based nonlinear optical crystal of 2-(2-(4-(4-chloro) phenyl) vinyl)-1, 1, 3-trimethyl-1H- benzo[e] Indolium iodide. The title crystal was grown by slow evaporation technique using methanol as a solvent. The harvested crystal is subjected to the single-crystal X-ray diffraction study and found that the title crystal belongs to the monoclinic system with a space group of P21/n. The bandgap and optical window of the material are studied by UV-visible spectroscopic analysis, and the found values are 4.2 eV and 295-800 nm, respectively. The grown crystal is subjected to analyze the thermal stability and found that high-temperature stability value upto 264 oC and which is sufficient for industrial applications. Furthermore, the effect of temperature on the grown crystal dielectric properties such as dielectric constant and dielectric losses is investigated by impedance spectroscopic analysis. The molecular charge transfer on the material is studied from density functional theory. The various intermolecular interactions existence in the crystal is explored from Hirshfeld surface analysis. Finally, the Z-scan parameters expose that title crystal is a potential material for third-harmonic generation applications. The overall structural studies and functional properties disclose that the grown crystal is well suitable for harmonic generation applications. Research Article Keywords: Crystal structure, DFT, Z-scan, Thermal analysis, Third harmonic generation Posted Date: December 29th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-1184837/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/20 Page 1/20 Page 1/20 1. Introduction Crystals have numerous uses in the present technology, notably in communication and laser technology, optical imaging, optical data storage, sensor, and nonlinear optics. Every day, there are new items have been made and new electronics arise from these crystals. With the advancement of the crystal growth technology of materials with thought-provoking nonlinear aspects, the technological world is looking for new materials. Controlling and considering the mechanisms of crystallization are important in current materials science, specifically it remains an even greater problem in the field of reticulate solids [1–3]. Nonlinear optics (NLO) is the interaction phenomenon with nonlinear crystals that controls the features of immensely coherent light. Advanced technological domains like optoelectronics and photonics depend on the NLO crystals as they are the mandatory functional materials in many applications. The substantial optical nonlinearity is one of the distinctive and crucial qualities of the NLO crystals, because of this attribute, it is being employed as the low profile cut-off frequency. This unique feature of the NLO crystals is employed in photosensitive electronics. Crystal growers have been made a significant contribution in the last couple of decays to find the high efficient organic NLO materials with outstanding optical characteristics and still, researchers are struggling a lot to find an outstanding material in all aspects such as functional properties as well as growth characteristics and hence still this research topic is considered as a challenging task. Organic crystals are bonded by weak van der Waals interactions and hydrogen bonds [4–6]. Because, the conjugated-bond system is the main framework in these organic NLO materials, and high mobility of the electron density is ascribed to the delocalized distribution of electronic charge owing to orbital overlap. Page 2/20 Page 2/20 Furthermore, the electron donor and acceptor groups on the ends of the bond systems may increase the asymmetric electronic distribution in both ground and excited states, increasing optical nonlinearity. Compared with the broad measure of examination led on the optical crystals for second-order nonlinear optical applications, the investigation of third-order nonlinear properties of atomic crystal has gotten moderately restricted consideration [7]. The interest in finding new third-order nonlinear optical materials, which are driven essentially by the search of third-order NLO materials for all-optical switching and sensor security applications, are worried about nonlinear absorption (NLA), nonlinear refraction (NLR), and third- order nonlinear susceptibility (χ3). 1. Introduction The phenomenon of molecular and crystal design techniques for third- order NLO applications is of incredible current interest. There have been impressive interest in organic optical material research because of a wide difference of plan prospects, faster nonlinearities that could be utilized for optical modulation, optical switching, and optical memory for emerging optical technologies [8, 9]. In any case, the enormous large macroscopic nonlinearity is needed for the applications in THz generation and detection and electro-optics. Additionally, requires different attributes like solubility, crystal growth, melting point, and thermal stability are also essential [10, 11]. Especially D-π∙∙∙A chromophore molecules have exhibited laser frequency conversion, optical communication, electro-optical modulation, THz generation, and detection [12–17]. Currently, developing this type of highly non-linear optical crystal is a challenging task. Therefore, we have selected the heterocyclic benzo [e] indole compounds that have D-π∙∙∙A chromophore. This heterocyclic benzo [e] indol was an electron acceptor and had a strong electron receptor. The large volume of benzo [e] indol increases the molecular asymmetry, which is highly efficient for nonlinear optical activities. Consequently, efforts were taken to synthesize and grown the new compound, 2-(2-(4-(4-chloro) phenyl) vinyl)-1, 1, 3- trimethyl-1H-benzo[e] Indolium iodide (CBI) single crystal. And, we have succeeded to synthesize and develop the best quality of CBI single crystal with full-scale size ~ 5 x 4 x 1.5 mm3 inside a period of 3 weeks. In this work, we have grown new benzo [e] indol CBI crystal with large third-order nonlinear properties. The grown crystal is further subjected to various characterization studies of Single-crystal XRD, Optical, Dielectric, and Thermal response of the sample, and the results are discussed to ensure the material properties towards the NLO applications. And the various intermolecular interactions that exist in the CBI crystal are explored from Hirshfeld surface analysis. To the best of our knowledge, the third-order NLO properties by Z-scan, dielectric, and Thermal studies of CBI crystals are reported for the first time. 2.1 Material synthesis and crystal growth The equimolar ratio of 1, 1, 2-Trimethyl-1H-benze[e] indole and Iodomethane were dissolved in toluene, and the mixture was refluxed. Then the precipitate was collected by filtration and washed five times with diethyl ether to afford the product as 1, 1, 2, 3-tetramethyl-1H-benzo [e] indol-3-ium-iodide (BI) [18]. The Page 3/20 Page 3/20 product BI and 4-chlorobenzaldehyde were dissolved in 50 ml of methanol and some amount of piperidine was also added and, which acts as a catalyst. The mixture was heated and refluxed for 18 hrs. After the refluxing, the mixture was cooled at ambient temperature. The final product of 2-(2-(4-(4-chloro) phenyl) vinyl)-1, 1, 3-trimethyl-1H-benzo[e] Indolium iodide (CBI) is formed as a solid with an orange color and the grown crystal shown in Fig. 1. 3. 2 Optical Studies For this study, a CBI solution was dissolved in methanol. Most Indolium compounds are soluble in methanol, since this causes the state to dissociate, resulting in free cations and anions. Fig. 4 depicts the measured optical absorption spectra of CBI. We can estimate the optical band gap and investigate the nature of transitions using absorption peaks. The spectrum shows a higher absorption peak at 295 nm, which corresponds to the n-π electronic transition. The calculated bandgap was to be 4.2 eV. Because of its outstanding optical behavior, absorption is not noticed between 295 and 800 nm, which is a benefit of CBI and an essential criterion for materials employed in nonlinear optical applications. 3.3 Thermal Stability Thermal stability of the CBI crystals characterized via differential thermal analysis (DTA) and thermal gravimetric (TG) measurements are shown in Fig. 5. The DTA and TG of the grown crystal were recorded in the range of 30-600 ⁰C under N2 atmosphere using the instrument Model Q600 SDT and Q20. As shown in Fig. 5, the sharpness of the endothermic peak at 264°C suggests the melting point of the CBI crystal. Interestingly, this is a higher melting temperature than that of benchmark organic NLO crystals of OH1 (212 ℃) and DAST (256 ℃). The CBI was stable up to 202 ⁰C, and also, the CBI exhibited very high initial weight loss in 246 ⁰C- 280⁰C for TG curves. The next stage of minimum weight loss occurs at 323 ℃ - 423 ℃. The following weight loss is the complete decomposition beyond the temperature of 438°C. Finally from this DTA-TG analysis, the thermal stability of the material is higher than the benchmark of nonlinear materials of DAST and OH1, and hence it can be exploited for optoelectronic device applications up to 264°C and suitable for high physicochemical stability applications. This advantage is also suitable for stable high-power Terahertz output. 3.1. Single-crystal X-ray diffraction analysis 1971339 Table 2 Hydrogen-bond geometry (Å, º) D‒H···A D‒H H···A D···A D‒H···A C23‒H23B···Cl1 0.96 3.81 2.961 148 C22‒H22C···Cl1 0.96 3.60 2.914 128 3.1. Single-crystal X-ray diffraction analysis Good quality single crystals of CBI were selected using an optical polarizing microscope for X-ray diffraction intensity data collection. The X-ray diffraction intensity data has been collected at room temperature from Bruker D8 QUEST ECO Single Crystal X-ray diffractometer equipped with MoKα radiation. The data collection has been monitored by the APEX III program suit. The structure was solved by the direct method with SHELXS2019 and refined by the full-matrix least-squares method with SHELXL2019 implemented in the WINGX2019 package[19, 20]. All H-atoms were located in different Fourier maps and refined isotopically. The molecule with thermal ellipsoid atoms plot was generated from the ORTEP3 incorporated in the WINGX2019 package. The molecular structure of the CBI has been determined from the single crystal x-ray diffraction analysis. Fig. 2 shows the 50% thermal ellipsoid plot of CBI molecule with the atom numbering scheme. Crystal properties, data collection, and structure refinement details are presented in Table 1. Further, the molecular structure of CBI has been determined from the gas-phase quantum chemical calculation. The selected geometrical parameters such as bond lengths, bond angles, and torsion angles of CBI molecule determined from the crystal phase and gas phase are collectively presented in Table S1. Comparing the structure of gas-phase and crystal phase, they are found to be equal and same (Fig. 2.); this can be well understood from their torsion angles. In the crystal phase, the intermolecular interactions are analyzed to understand the supramolecular properties. The crystal packing is stabilized by C‒H‧‧‧H and π‧‧‧π type of interactions. The molecules are linked by C23‒H23B‧‧‧Cl1 and C22‒H22C‧‧‧Cl1 interactions. The detailed intermolecular interactions with the symmetry code were tabulated in Table 2 and Fig. 3 shows the intermolecular interactions view along the 010 axes and the Crystallographic data for CBI (CCDC Number - 1971339) have been deposited with the Cambridge Crystallographic Data Centre. Page 4/20 Table 1 Crystal Structure details Crystal Structure details Chemical Formula C23H21NCl I Formula Weight (g/mol) 473.76 Crystal System Monoclinic Space Group P21/n a/Å 7.2849 (4) b/Å 21.9014 (13) c/Å 12.4896 (8) α/(o) 90.00 β//(o) 96.359 (2)° γ//(o) 90.00 V/Å3 1992.7 (2) Z 4 T/K 293 R1 0.021 Radiation type Mo Kα µ (mm−1) 1.75 Crystal size (mm3) 0.51× 0.21 × 0.15 Refinement R[F2 > 2σ(F2)], wR(F2), S 0.037, 0.087 No. of reflections 3171 No. of parameters 386 H-atom treatment H-atom parameters constrained Δρmax, Δρmin (e Å−3) 0.86 −1.31 CCDC No. 3.4 Dielectric analysis By the way, in the NLO materials research, the investigation of temperature depends on dielectric properties, and its stability concerning temperature is highly crucial. The degree of the polarization greatly contributes to the harmonic generation of the crystal [21–23]. Based on the technological importance, we have performed impedance spectroscopic studies using an impedance analyzer to understand the test specimen's electronic polarization. The calculated dielectric constant and dielectric loss profiles over the frequency region 1 Hz to 1MHz concerning various temperature are provided in Fig. 6 (a) and Fig. 6 (b). As seen in Fig. 1, at 40°C the constant dielectric profile showed the universal behavior of the previous NLO Page 6/20 materials concerning the frequency, and it illustrated that the four types of polarizations are contributed [24, 25]. Furthermore, we increased the temperature to 60, 80, and 100°C and obtained constant dielectric profiles are presented in Fig. 6 (a) and Fig. 6 (b). Surprisingly, we could not see any changes in the dielectric constant values, and the dielectric constant trend is also similar to all the other temperatures. Hence, it is clear that the electronic polarization of the test sample with respect to temperature is not affected, and the applied temperature has not induced any surface and structural defects, and the resultant output of the harmonic generation is not affected. Note that such electronic polarization stability is much required for the electronic devices fabrications to save the device by the external parameters [26]. In addition to that, based on previous reports, numbers of NLO materials have been undergoing significant changes in the constant dielectric profiles concerning temperature. Herewith, few reports are discussed based on their changes in the dielectric constant profiles to temperature for better understanding the importance of the stability of the dielectric properties. Sangeetha et al., have been reported glycinium hydrogen fumarate glycine solvate monohydrate single crystal dielectric constant profiles concerning temperature and found massive changes in the dielectric constant and loss [27]. L-Threonine formate crystal dielectric constant profiles with respect to temperatures have been reported by Redrothu Hanumantharao et al. and found considerable changes in the dielectric constant with respect to temperature [28] and similar kind of results are also found in L-Asparaginium picrate [29] and L-Argininium perchlorate [30]. Nevertheless, in the present case, the tilted crystal does not show considerable change with respect to temperature. 3.4 Dielectric analysis Hence, the titled material is highly deserved for the device fabrications under different temperatures. 3.6 Frontier Molecular Orbital analysis Geometry optimization of the synthesized molecule (CBI) was carried out by Becke’s three-parameter hybrid exchange functional (B3LYP) level of density functional theory (DFT). All the atoms were optimized by the LANL2DZ basis set to get more accurate energy optimization of the molecule, all the gas phase structural optimization and computation were performed by the Gaussian09 software package. The higher occupied molecular orbital (HOMO), lower unoccupied molecular orbital (LUMO) have been analyzed from Gauss view package [35]. The essential quantum chemical descriptors such as highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) were calculated for the imperator in the molecule, which is highly related to the chemical and biological activities. The molecular surface of HOMO and LUMO maps of both complexes were drawn at 0.2 au and shown in Fig.9, in which the HOMO and LUMO energy surfaces were found over the keto group and centre ring regions, respectively. The positive and negative regions are shown in green and red colors, respectively. The global reactivity descriptors were calculated to understand the quantum chemical properties. The calculated values for the HOMO and LUMO energies are -4.76eV and -3.16eV, respectively. The bandgap is 1.60eV. The calculated electron affinity, ionization potential, and electronegativity are 3.16, 4.76, and 3.96eV. The electron affinity and electronegativity values are low compared to the ionization potential, which suggests the molecule has high reactivity. 3.5 Hirshfeld surface analysis Hirshfeld surface analysis is used to visualize and quantify various types of intermolecular interactions in the crystal environment. Also, it is used to define the partitioning of electron density of the crystal into molecular fragments[31, 32]. Intermolecular contacts and their impact on crystal packing were analyzed using Crystallographic Information File (CIF) in Crystal Explorer 17.5 software [33, 34]. The de (outside nuclei surface) and di (inside surface) were scaled in different colors and displayed the sum in Fig. 7(a) and 7(b) (vdw radii in white, close-contact interactions in red and short contacts than vdw radii in blue surfaces). Further, the shaped index explores the C−H∙∙∙π and π∙∙∙π interactions. Fingerprint plots used to determine the contact distances (di, de) and the reciprocal contacts are in the range from 0.6 to 2.8 Å. In the dnorm plot (Fig. 7), white surface is the sum of vdw radii, the blue surface is the short contact distances than vdw radii and red surface explains the hydrogen bond contacts. Further, the intermolecular interactions of the molecule in the crystal can be characterized by the 2D shape of fingerprint plots shown in Fig. 8. The figure displays the fingerprint plot, which helps quantify the contribution of different types of interactions in the solid-state. The H···H, O···H and C···H type of interactions are highly presented in CBI, which confirm that the Vander Walls and C‒H···π interactions play the major role in the crystal environment. The C···H fingerprint plots show similar symmetrical wing shape, the O···H plot looks like sharp spikes and the H···H plot are overlapped in full fingerprint plots that Page 7/20 Page 7/20 would commonly separate from each other. Therefore, this fingerprint plot confirms that weaker hydrogen bonding leads to the stability of the compound in the crystal phase. 3.7 Nonlinear optical property The third-order NLO properties of CBI were studied using the open and closed aperture Z-scan technique [36]. It is far a sensitive and straightforward method to measure the optical nonlinearities with high accuracy. This method focuses on quantifying NLA (nonlinear absorption) and NLR (nonlinear refraction) simultaneously with a single experiment. It gives the most helpful information about the sign and magnitude of the obtained nonlinearities. Open aperture (OA) Z-scan is used to determine NLA and closed aperture (CA) Z-scan to Measure NLR. In this current work, we achieved Z-scan measurements with a diode-pumped solid-state (DPSS) continuous wave (CW) laser of Gaussian beam profile with 532 nm and 200 mW output power. The laser beam becomes tightly targeted, with the Plano-convex lens having a 28.6 cm focal length. The beam size and the depth of the laser at the focus point are measured to be 33 µm and 12.8 kW/cm2, respectively. The 1 mm cuvette containing the CBI solution was placed on a stepper motor (100 µm resolution) and scanned across the lens's focus along the Z-axis path (laser beam propagation). The Z-scan measurement of the thin medium situation is fulfilled because sample thickness is much less than the Rayleigh length (Z0= 6.15 mm). The cross-section of the laser beam coming out of the sample turned into controlled to placing an aperture of variable diameter within a far- field discipline. Whereas only in part transmitted intensity is measured in closed aperture experiment, and the aperture size became kept at 40% of the linear transmittance (S = 0.4). The detectors had been connecting to the virtual electric control unit, the interpretation stage and data acquisition changed into control with the aid of the automatic Z-scan program Page 8/20 Figure 10: Nonlinear absorption curves of CBI Figure 10: Nonlinear absorption curves of CBI 3.7.1 Nonlinear absorption It is determined that CBI suggests a lower transmittance with an increase in intensity, indicating the reverse saturable absorption (RSA) behavior, i.e., the absorption is maximum at the focus, wherein the intensity is maximum (Z=0). Therefore, a signal of the nonlinear absorption is positive for RSA (β > 0). In order to estimate the nonlinear absorption coefficients quantitatively and affirm the corresponding NLA mechanism, we equipped the experimental open aperture data with the theoretical NLA model. The stable lines within Fig. 10 represent the theoretical curves, and open circles are experimental information. The Two-photon absorption (TPA) kind of nonlinear absorption becomes determined to be the fine approximation. The normalized transmission as a feature of the sample position is given by [21]. T(Z) = 1 − βI0Leff 2√2(1 + Z2/Z2 0) 1 Where β is the two-photon absorption coefficient, which is found to be within the order ~10−4 cmW−1 for all samples. Leff = (1 −exp −aL)/a, is the effective length of the sample, α is the linear absorption coefficient, and L is the thickness of the sample. I0 is the intensity of the laserbeam at the focus. The corresponding β values of the samples are supplied in Table 3. Where β is the two-photon absorption coefficient, which is found to be within the order ~10−4 cmW−1 for all samples. Leff = (1 −exp −aL)/a, is the effective length of the sample, α is the linear absorption coefficient, and L is the thickness of the sample. I0 is the intensity of the laserbeam at the focus. The corresponding β values of the samples are supplied in Table 3. 3.7.2 Nonlinear refraction The nonlinear refraction curves of the CBI are shown in Fig. 11. We observed negative nonlinear refraction behavior for both the samples due to the self-focusing effect of the samples. The nonlinear phase shift of the samples was obtained by fitting the closed aperture data with the following relation [37]. T(Z) = 1 − (4X? ? 0) (X2 + 1)(X2 + 9) 2 where T (Z) is the normalized transmittance, Δ\upvarphi0 is the nonlinear phase shift, and X = Z/Z0. Nonlinear refractive index n2 can be calculated by using the Δ\upvarphi0value in the relation ? ? 0 = kn2I0Leff, where k (=2π/λ) is the wave vector. The obtained values of the NLR coefficients are found to be in the order ~10−8 cm2W−1 and are given in Table 3. The third-order nonlinear refraction and absorptions are directly related to the real and imaginary parts of the third-order nonlinear optical susceptibility, respectively. The real, imaginary, and magnitude of the susceptibility of the CBI and HBI have been decided with using the following relations [38]., Page 9/20 Page 9/20 ? ( 3) = ? ( 3) R + i? ( 3) I 3 ? ( 3) R (esu) = cn2 0 120p2n2(m2/W) 4 ? ( 3) R (esu) = cn2 0 120p2n2(m2/W) 4 χ ( 3) I (esu) = c2n2 0 240π2ω β(m/W) 5 Where c is the velocity of light, n0 is the linear refractive index and ω is the angular frequency of the incident photons. The obtained magnitude of the NLO susceptibility of CBI is in the order ~10−7 esu. Further, the molecular second hyperpolarizability (microscopic polarizability) of the samples was also calculated and the values are found to be in the order ~10−26 esu, indicates the acquired values of nonlinear optical parameters from Z scan measurements of CBI crystal. The comparison of third-order optical susceptibility of CBI crystal with a few well known organic crystals is given in Table.4. The grown CBI has a very high susceptibility, which also increases the capability of the material in NLO tool fabrication. 3.7.2 Nonlinear refraction Table 3 Nonlinear optical parameter for CBI Crystals Δϕ 0.45 Nonlinear absorption coefficient (β) 4.80x 10−4 (cm/w) Nonlinear refractive index (n2) -0.33x 10−8 (cm2/w) Real part of third order susceptibility (Re χ(3)) -1.74 x 10−7 esu The imaginary part of third-order susceptibility (Imχ(3) ) 1.08 x 10−7 esu Third-order nonlinear susceptibility (χ(3)) 2.05 x 10−7 esu Second order molecular hyperpolarizability (γh) 0.97 x 10−26 esu Page 10/20 Page 10/20 Table 4 Comparison of χ3 value of CBI with some other NLO single crystals S. No Crystals Third-order optical susceptibility (χ3) References 1. VMST 9.69 x 10−12 esu [39] 2. 2A5NPBr 6.76×10−09 esu [40] 3. DSMOS 5.05×10−8 esu [41] 4. 3ETSI 3.84×10−7 esu [42] 5. CBI 2.05 x 10−7 esu Present Work Table 4 4. Conclusion The new organic compound of 2-(2-(4-(4-chloro) phenyl) vinyl)-1, 1, 3-trimethyl-1H-benzo[e] Indolium iodide was synthesized and the crystal was grown by slow evaporation technique. The structural features are established from a single-crystal X-ray diffraction analysis. As per the single XRD data, the CBI crystal belongs to the monoclinic crystal system with the centrosymmetric symmetry space group of P21/n. The optical absorption study reveals that the lower absorption in the visible spectral region is a good fit for the NLO applications. The material is thermally stable upto 264 oC, which is higher than the typical DAST crystal. The molecular level charge transfer of the material was studied from the frontier molecular orbital analysis. At various temperatures, the fluctuation in the dielectric constant was investigated. Z-scan with a closed aperture showed the positive nonlinear absorption coefficient. 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Sagayaraj, Investigation on third-order nonlinear optical, electrical, and surface properties of organic stilbazolium crystal of 4-N,N-dimethylamino-N′-methylstilbazolium p- methoxybenzenesulfonate, Opt. Mater. (Amst). Figures Figure 1 Photograph of as grown CBI crystals Figure 1 Photograph of as grown CBI crystals Page 15/20 Page 16/20 Figure 2 Ortep view of CBI molecule Figure 3 Figure 2 Ortep view of CBI molecule Figure 3 Page 16/20 Page 16/20 Intermolecular interactions view along 010 axis Figure 4 Optical absorption spectra of CBI Fi 5 Figure 6 (a) Dielectric constant with respect to temperature and (b) Dielectric Loss with respect to temperature (a) Dielectric constant with respect to temperature and (b) Dielectric Loss with respect to temperature Figure 4 Optical absorption spectra of CBI Figure 5 TGA and DTA analysis of CBI TGA and DTA analysis of CBI h respect to temperature and (b) Dielectric Loss with respect to temperature Page 17/20 Page 17/20 Figure 7 Hirshfeld surface of a) dnorm structure b) shape index structure of CBI molecule Figure 7 Page 18/20 Figure 8 Figure 8 Page 18/20 Page 18/20 Fingerprint plots of CBI Figure 9 Th HOMO d LUMO h d t 0 02 l l Figure 11 Figure 11 Nonlinear refraction curves of CBI Figure 9 Figure 9 The HOMO and LUMO has drawn at 0.02 au level The HOMO and LUMO has drawn at 0.02 au level Page 19/20 Page 19/20 Page 19/20 Figure 10 Supplementary Files This is a list of supplementary files associated with this preprint. Click to download. SupplymentryTableS1.docx Page 20/20 Figure 10 Nonlinear absorption curves of CBI Nonlinear absorption curves of CBI Nonlinear absorption curves of CBI
https://openalex.org/W2795010735	https://aip.scitation.org/doi/pdf/10.1063/1.5017801	English	null	Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal	AIP advances	2,018	cc-by	4,749	RESEARCH ARTICLE \| MARCH 29 2018 Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal RESEARCH ARTICLE \| MARCH 29 2018 Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal RESEARCH ARTICLE \| MARCH 29 2018 Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal Motoki Kunihiro; Yasuaki Nakasone; Shofu Matsuda; Sayoko Shironita; Norio Nagayama; Minoru Umeda Motoki Kunihiro; Yasuaki Nakasone; Shofu Matsuda; Sayoko Shironita; Norio Nagayama; Minoru Umeda AIP Advances 8, 035324 (2018) https://doi.org/10.1063/1.5017801 AIP Advances 8, 035324 (2018) https://doi.org/10.1063/1.5017801 Articles You May Be Interested In Interfacial reactions between indium tin oxide and triphenylamine tetramer layers induced by Interfacial reactions between indium tin oxide and triphenylamine tetramer layers induced by photoirradiation photoirradiation J. Appl. Phys. (May 2008) J. Appl. Phys. (May 2008) Observation of hole injection boost via two parallel paths in Pentacene thin-film transistors by employing Pentacene: 4, 4″-tris(3-methylphenylphenylamino) triphenylamine: MoO3 buffer layer APL Mater. (November 2014) Solvent-induced symmetry-breaking charge transfer in an octupolar triphenylamine derivative resolved with transient fluorescence spectroscopy 24 October 2024 05:55:29 AIP ADVANCES 8, 035324 (2018) Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal Crystal structure oriented carrier transport characteristic of triphenylamine derivative single crystal Motoki Kunihiro,1 Yasuaki Nakasone,1 Shofu Matsuda,1 Sayoko Shironita,1 Norio Nagayama,1,2 and Minoru Umeda1,a 1Nagaoka University of Technology, Kamitomioka, Nagaoka, Niigata 940-2188, Japan 2Ricoh Company, Ltd., Izumi, Ebina, Kanagawa 243-0460, Japan (Received 30 November 2017; accepted 20 March 2018; published online 29 March 2018) Organic electronic materials used in organic photoreceptors for electrophotography areexpectedtobeemployedinmanyfunctionalorganicdevices.Regardingtheinferior mobility compared to inorganic semiconductors, in this study, an organic single crys- tal was prepared at a high rate and characterized. First, solubility and supersolubility curves of α-phenyl-4’-[(4-methoxyphenyl)phenylamino]stilbene (TPA) were mea- sured. According to the data, the rapid crystal growth has been succeeded. Polarization microscope observation and XRD result conﬁrmed that a single crystal is obtained. The results of a cyclic voltammogram and molecular orbital calculations showed that hole transport takes place in the crystal. On the surface of the obtained single crystal, Au parallel electrodes were vacuum deposited to measure its electric conductivity. Higher currents were observed on the long and short axes of the single crystal than that of the amorphous layer. Moreover, an anisotropic conductivity, in which the cur- rent in the long axis is greater than that in the short axes, was remarked. Consequently, the molecular stack of the crystal clariﬁed by an XRD analysis and molecular elec- tronic structure assessed by molecular orbital calculation well explain the anisotropic conductivity of the crystal. © 2018 Author(s). All article content, except where oth- erwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). https://doi.org/10.1063/1.5017801 24 October 2024 05:55:29 aElectronic mail: mumeda@vos.nagaokaut.ac.jp (Corresponding author) aElectronic mail: mumeda@vos.nagaokaut.ac.jp (Corresponding author) 2158-3226/2018/8(3)/035324/7 8, 035324-1 © Author(s) 2018 I. INTRODUCTION Organic photoreceptors for electrophotography are abundantly used in practical applications of laser-beam printers and digital copiers.1 One attractive feature of the organic device is the high light- to-electrical energy conversion efﬁciency of >0.95,2–4 which is sufﬁcient to be implanted into organic photovoltaics (OPVs) to achieve a high conversion efﬁciency. Irrespective of the remarkable high efﬁciency of the photoreceptors, other organic photo-sensing devices are not actually utilized.5–16 This is because the signiﬁcant success of the photoreceptors is based on a speciﬁc electrophotography process in which the slow time response for the image formation is required to be ∼300 ms. Organic electronic materials, developed from the organic photoreceptors, are expected to be employed in many functional organic devices.17–24 Although much effort has been spent for realiza- tion, only organic light emitting diodes are actually used in practice.5–8,17,19,24,25 The main reason Organic electronic materials, developed from the organic photoreceptors, are expected to be employed in many functional organic devices.17–24 Although much effort has been spent for realiza- tion, only organic light emitting diodes are actually used in practice.5–8,17,19,24,25 The main reason behind the small number of successes in the organic devices is attributed to the low carrier mobility of these materials. In the electrophotographic image formation process,2 the carrier mobility of the organic photoreceptors as high as ∼10-5 cm2V-1s-1 is not very slow,26 whereas a high carrier mobility is needed to achieve functional organic devices.17–24 tion, only organic light emitting diodes are actually used in practice. , , , , The main reason behind the small number of successes in the organic devices is attributed to the low carrier mobility of these materials. In the electrophotographic image formation process,2 the carrier mobility of the organic photoreceptors as high as ∼10-5 cm2V-1s-1 is not very slow,26 whereas a high carrier mobility is needed to achieve functional organic devices.17–24 g In the above-mentioned organic devices, amorphous forms are usually used.5–25 For the speciﬁc improvement of the carrier mobility of organics, crystallization seems to be a successful method.27–30 The ordinary stacked continuum structure in a crystal accelerates the carrier transport between the aElectronic mail: mumeda@vos.nagaokaut.ac.jp (Corresponding author) 2158-3226/2018/8(3)/035324/7 8, 035324-1 © Author(s) 2018 2158-3226/2018/8(3)/035324/7 035324-2 Kunihiro et al. AIP Advances 8, 035324 (2018) nearest-neighbor molecules and ensures a negligibly low number of carrier-trap sites. For realizing some organic devices, an organic single crystal with a large size should be prepared by developing a high growth rate. I. INTRODUCTION Previously, by employing a carrier transport molecule used in an organic photoreceptor,31 we reported the high-rate preparation procedure of a large single crystal.32 As a result, a higher car- rier transport was found in the direction in which the intramolecular distance is smaller than that of the larger one. Interestingly, the organic crystal structure is reported to be transformed for the triphenylamine-derivative molecule by a small change in the chemical structure.33 Thus, we focused our attention on another triphenylamine molecule which is reported to have a different crystal struc- ture than the previously reported molecule.32 In the present study, the single crystal preparation of α-phenyl-4’-[(4-methoxyphenyl)phenylamino]stilbene (TPA) is described. The relationship between its molecular structure and carrier transport property is then discussed. Finally, the anisotropic carrier transport based on the crystal structure is clariﬁed. D. Single crystal analysis and anisotropic carrier transport The crystal structure was evaluated by an XRD instrument (Bruker AXS) equipped wit monochromatized Mo-Kα radiation (λ = 0.71069 Å). A pair of parallel Au electrodes was fabricated on the TPA crystal surface by vacuum evaporation (ULVAC, VPC-260F) using a metal mask. The size of Au electrodes is 2-mm length and 1-mm width, and its interval is 20-µm. Two pair of parallel electrodes were deposited on the TPA crystal surface in the directions of the long-axis (c axis) and short-axes (a and b axes). An amorphous TPA layer was fabricated on a glass plate with a 50 wt% THF solution for comparison. After drying the recast layer, the Au electrodes were fabricated in the same manner. The current-voltage curve of the TPA was measured in a temperature-controlled chamber (Collet- Kogyo Co.) equipped with a pair of tungsten probes and a system source meter (Keythley, 2612A). Considering the level of TPA resistance, we employed the two-probe measurement method. The tungsten probes were contacted to the Au electrodes. The current was measured between the Au electrodes at 28.8◦C and 1x10-3 Pa by scanning the applied voltage from 0 to +200 V at the scan rate of 0.2 Vs-1 under a dark condition. Also, the measurement of the current-voltage property was performed for the TPA crystal at the temperatures of 48.8, 68.8, and 88.8◦C to construct an Arrhenius plot. C. Molecular orbital calculation and electrochemical measurement The molecular orbital calculation of TPA was performed by the computational method using the software program WinmostarTM (X-Ability Co.,Ltd.). For its structural optimization, a package of General Atomic and Molecular Electronic Structure System (GAMESS) was used. For the electrochemical characterization, 5 mmol·dm-3 TPA dissolved in an acetonitrile (AN) solution containing the supporting electrolyte of 0.1 mol·dm-3 tetraethylammonium perchlorate (TEAP) was prepared. A Pt disk electrode of 300-µm diameter and Pt coil were used as the working and counter electrodes, respectively. A Ag/AgCl was used as a reference electrode. All reagents were electrochemical-grade and used without further puriﬁcation. N2 gas (99.9999%) bubbling purges into the electrolyte solution before the cyclic voltammetry measurements. The voltammetry mea- surement was performed using a potentiostat (BAS, ALS660A). The scan rate was from 0.05 V·s-1 to 0.5 V·s-1. 24 October 2024 05:55:29 B. Optical observations and polarization feature By a utilization of an optical microscope (Moritex, Inf-500-DA), a photograph of the crystal prepared by the above-mentioned method was taken. The crystal size was measured using a laser scanning microscope (Olympus, OLS-1200). Using a polarization microscopy (Nikon, OPTIPHOT- 100) in the reﬂection mode, the polarization features of the obtained crystal were clariﬁed. A. Preparation of single crystal In the insert of Fig. 1, the molecular structure of the TPA employed in this study is seen. For the measurements of solubility curve and supersolubility curve of the TPA in a tetrahydrofuran (THF) solvent, a screw-capped glass tube including 40 wt% TPA and 60 wt% THF was immersed in a water bath (Julabo, F25-ME). The temperature of the glass tube was then steadily raised by controlling that of the water bath at a rate of 4 ◦C/h until the TPA powder had clearly dissolved in the solution so that the dissolution temperature can be determined. After that, the tempera- ture of the glass tube was slowly cooled by controlling the water bath temperature at the rate of 3 ◦C/h for regenerating the TPA solid. The regenerating point was noted as of the crystallization temperature. Then, the TPA content of the solution was altered to 45, 50 and 55 wt% to ﬁnd the temperatures of dissolution and crystallization similarly as mentioned above. By connecting the dissolution and crystallization temperatures, supersolubility and solubility curves were recognized, respectively. p y Based on the above-observed solubility and supersolubility data, a single crystal was obtained as follows. The screw-capped glass tube including a 45 wt % TPA solution was immersed in a 35◦C- controlled water bath. Subsequently, a seed crystal of the TPA prepared in advance was put into the FIG. 1. Solubility and supersolubility curves of the TPA-THF solution. The crystal preparation condition is chosen between the two curves. The inset shows the molecular structure of α-phenyl-4’-[(4-methoxyphenyl)phenylamino]stilbene (TPA) used in this study. FIG. 1. Solubility and supersolubility curves of the TPA-THF solution. The crystal preparation condition is chosen between the two curves. The inset shows the molecular structure of α-phenyl-4’-[(4-methoxyphenyl)phenylamino]stilbene (TPA) used in this study. 035324-3 Kunihiro et al. AIP Advances 8, 035324 (2018) 35◦C TPA solution. The temperature of the glass tube was then slowly cooled to 29◦C at the rate of 0.125 ◦C/h for 48 hours, resulted in the growth of the TPA crystal. Throughout this procedure, the TPA solution was kept in the dark. 35◦C TPA solution. The temperature of the glass tube was then slowly cooled to 29◦C at the rate of 0.125 ◦C/h for 48 hours, resulted in the growth of the TPA crystal. Throughout this procedure, the TPA solution was kept in the dark. A. Preparation of single crystal By using the TPA-dissolved THF solutions, the measured solubility and supersolubility curves are shown in Fig. 1. The preparation of a TPA crystal was conducted at the area of between the solubility and supersolubility curves in Fig. 1. The growth of TPA crystal was carried out in a 45 wt.% TPA solution at the cooling rate of 0.125◦C/h starting from 35◦C for 48 h. Consequently, a crystal of 8 x 0.8 x 0.4 mm was obtained as shown in Fig. 2. The speciﬁc gravity of crystal was 1.05 based on its weight and volume. Fig. 3 displays images taken by the open-Nicole and cross-Nicole optical paths by changing the angle of the goniometer bearing the crystal using a reﬂection polarizing microscope. Figs. 3A and C are recorded under the open-Nicole optical path. Figs. 3B and D were taken under the condition of 035324-4 Kunihiro et al. AIP Advances 8, 035324 (2018) FIG. 2. Optical photograph of the prepared TPA crystal. The size is 8 x 0.8 x 0.4 mm. FIG. 3. Reﬂection polarization microscopy images of the TPA crystal. A and C represents the open-Nicole micrographs. B and D are the brightest and darkest taken under the cross-Nicole optical path, respectively. An extinction is observed in D. 035324-4 Kunihiro et al. AIP Advances 8, 035324 (2018) FIG. 2. Optical photograph of the prepared TPA crystal. The size is 8 x 0.8 x 0.4 mm. FIG. 2. Optical photograph of the prepared TPA crystal. The size is 8 x 0.8 x 0.4 mm. FIG. 3. Reﬂection polarization microscopy images of the TPA crystal. A and C represents the open-Nicole micrographs. B and D are the brightest and darkest taken under the cross-Nicole optical path, respectively. An extinction is observed in D. 24 October 2024 05:55:29 FIG. 3. Reﬂection polarization microscopy images of the TPA crystal. A and C represents the open-Nicole micrograph B and D are the brightest and darkest taken under the cross-Nicole optical path, respectively. An extinction is observed in D the cross-Nicole optical path in which the picture angles are the brightest and darkest, respectively. At the same time, two goniometer angles of B and D were taken, and the difference in the angles was found to be 45 degrees. Especially, in Fig. 3D, an extinction of the objective is observed. These results ensured that the TPA single crystal was prepared. B. Chemical structure and carrier transport property Ag/AgCl without a corresponding reduction current, implying that the reaction is irreversible. Fig. 5B shows cyclic voltammograms at the scan rates from 0.05 to 0.5 Vs-1 to evaluate the ﬁrst oxidation wave.35 A plot of the ﬁrst oxidation peak current versus the square root of the scan rate is shown in the inserted ﬁgure. As a result, there is a linear relationship. It was found that the TPA in the solution is redoxed by the outer-sphere electron transfer. It is noted that no redox current peaks are observed at the potential range from -1.5 to 0.0 V vs. Ag/AgCl in the cyclic voltammogram of TPA. Based on the results of Fig. 5, the ﬁrst oxidaton potential of the TPA was determined to be Eox=0.756 V vs. Ag/AgCl. 24 October 2024 05:55:29 Considering that the ﬁrst oxidation potential corresponds to the energy level of HOMO,36 th ionization potential (IP) is determined as following:37 IPHOMO = Eox + 4.5 (eV), (1) (1) where Eox is expressed versus the standard hydrogen electrode (SHE). Hence, the IPHOMO of the PA is calculated to be 5.500 eV. where Eox is expressed versus the standard hydrogen electrode (SHE). Hence, the IPHOMO of the TPA is calculated to be 5.500 eV. Based on these results, the oxidized TPA is suggested to show the stable hole conduction in organic photoconductors actually used in printing systems. The oxidized TPA would play a role as a hole in the solid device, which corresponds to its radical cation that localizes at the triphenylamine group of the TPA molucule.38 B. Chemical structure and carrier transport property To analyze the electronic state of the HOMO, we conducted a molecular orbital calculation of the TPA single molecule. The result is shown in Fig. 4. The electronic state distribution was found to localize at the triphenylamine group, which suggests that the hole transport mainly occurs around the triphenylamine unit between the TPA molecules. Fig. 5 shows cyclic voltammograms of the TPA using the Pt disk electrode with the diam- eter of 300 µm in an acetonitrile solution. In Fig. 5A, the ﬁrst oxidation current is observed at FIG. 4. The molecular orbital of TPA single molecule at the energy level of the HOMO. FIG. 4. The molecular orbital of TPA single molecule at the energy level of the HOMO. AIP Advances 8, 035324 (2018) Kunihiro et al. FIG. 5. Cyclic voltammograms of 5 mmol·dm-3 TPA using φ300 µm Pt disk electrode in an acetonitrile solution containing tetraethylammonium perchlorate. (A) Scan rate: 0.05 Vs-1. (B) Scan rate: 0.05 – 0.5 V·s-1. FIG. 5. Cyclic voltammograms of 5 mmol·dm-3 TPA using φ300 µm Pt disk electrode in an acetonitrile solution containin tetraethylammonium perchlorate. (A) Scan rate: 0.05 Vs-1. (B) Scan rate: 0.05 – 0.5 V·s-1. ∼0.76 V vs. Ag/AgCl along with a reduction current, which is indicative of the reversible electrode reaction.34 On the other hand, the second split oxidation current is observed between 1.2 ans 1.5 V vs. Ag/AgCl without a corresponding reduction current, implying that the reaction is irreversible. Fig. 5B shows cyclic voltammograms at the scan rates from 0.05 to 0.5 Vs-1 to evaluate the ﬁrst oxidation wave.35 A plot of the ﬁrst oxidation peak current versus the square root of the scan rate is shown in the inserted ﬁgure. As a result, there is a linear relationship. It was found that the TPA in the solution is redoxed by the outer-sphere electron transfer. It is noted that no redox current peaks are observed at the potential range from -1.5 to 0.0 V vs. Ag/AgCl in the cyclic voltammogram of TPA. Based on the results of Fig. 5, the ﬁrst oxidaton potential of the TPA was determined to be Eox=0.756 V vs. Ag/AgCl. ∼0.76 V vs. Ag/AgCl along with a reduction current, which is indicative of the reversible electrode reaction.34 On the other hand, the second split oxidation current is observed between 1.2 ans 1.5 V vs. C. Single crystal analysis and anisotropic carrier transport Then, the crystal structure of the TPA was analyzed by XRD. The result and the lattice constants are shown in Fig. 6A. These results corresponded to the previous report where the crystal was prepared FIG. 6. (A) Single crystal X-ray diffraction result of the TPA crystal represented in Figs. 2 and 3 and the corresponding lattice constants. (B) A schematic diagram of the stacking structure of the TPA molecules. FIG. 6. (A) Single crystal X-ray diffraction result of the TPA crystal represented in Figs. 2 and 3 and the corresponding latti constants. (B) A schematic diagram of the stacking structure of the TPA molecules. AIP Advances 8, 035324 (2018) 035324-6 Kunihiro et al. Kunihiro et al. FIG. 7. (A) Schematic image of the differently oriented Au contacts on the TPA single crystal and (B) i-E characteristics of the TPA single crystal (a-, b-, and c-axes) at the temperature of 28.8◦C by comparison to that of the amorphous TPA layer. FIG. 7. (A) Schematic image of the differently oriented Au contacts on the TPA single crystal and (B) i-E characteristics of the TPA single crystal (a-, b-, and c-axes) at the temperature of 28.8◦C by comparison to that of the amorphous TPA layer. in the methanol solvent,33 which indicates that the TPA single crystal is successfully obtained. From the XRD analysis, the molecular stack structure can be estimated and is shown by the schematic representation in Fig. 6B. The stack degree of electron cloud in the c-axis is suggested to be greater than that in the a- and b-axes. Because the hole is transported among the neighboring TPA cation radicals, the degree of the electron overlapping is signiﬁcant for hole transport. As mentioned in the experimental section, the 20-µm gapped parallel Au electrodes were deposited on the TPA-crystal surface in the a-, b- and c-axes directions and on the amorphous layer. Fig. 7 shows the electric-ﬁeld dependence of the currents of the TPA single crystal and the amor- phous form measured at 28.8◦C. The c-, a-, and b-axes currents, normalized by the electrode length of the TPA crystal, is signiﬁcantly higher than that of the amorphous TPA, which is indicative of the high electric conductivity of the single crystal. C. Single crystal analysis and anisotropic carrier transport It should be noted here that the electric conduc- tivity in the c-axis is clearly higher than that in the a- and b-axes, suggesting that the anisotropic hole transport occurs in the single crystal. Considering the results of the molecular orbital calcula- tions (Fig. 4) and the XRD-based structural analysis (Fig. 6), the orientation of the triphenylamine group, where the electrons of HOMO localize, is suggested to induce the anisotropic conductiv- ity. Hence, the extent of the electron overlapping in the c-axis is larger than that in the a- and b-axes. Furthermore, we investigated the temperature dependence of the current-voltage property of the c- and b-axes of the TPA single crystal. Based on the result in the electric ﬁeld of 10 V/µm, we constructed the Arrhenius plot, then calculated the activation energy (Ea) and the pre-exponential factor (I0) in both axes using the following equations: I = I0e −Ea RT (2) ln I = ln I0 + −Ea R ! 1 T ! (3) (2) (3) where I, R, and T are the current, gas constant, and temperature, respectively. As shown in Table I, although I0 is almost same between the c-axis and b-axis, the Ea of c-axis is one half that of the b-axis. This indicated that the hole can be easily transferred in the direction of the c-axis, which strongly supports the anisotropic conductivity of the TPA single crystal as shown in Fig. 7. TABLE I. Activation energy (Ea) and pre-exponential factor (I0) of the c-axis and b-axis of the TPA single crystal. c-axis b-axis Ea 1.74 kJ/mol 3.49 kJ/mol 0.018 eV 0.036 eV I0 1.26×10-7 1.14×10-7 TABLE I. Activation energy (Ea) and pre-exponential factor (I0) of the c-axis and b-axis of the TPA single crystal. 035324-7 Kunihiro et al. AIP Advances 8, 035324 (2018) AIP Advances 8, 035324 (2018) IV. CONCLUSIONS For a speciﬁc improvement of the carrier mobility in hole-transport materials for organic devices, a single crystal of α-phenyl-4’-[(4-methoxyphenyl)phenylamino]stilbene (TPA) was prepared and its structural characteristics and carrier transport properties were evaluated in this study. A large-scale crystal of TPA was successfully prepared by a solution method based on the obtained solubility and supersolubility curves. The TPA crystal was observed by an optical microscope and a polarization microscope, and found that the obtained structure was a single crystal. The ionization potential of the HOMO was determined from the cyclic voltammogram of the TPA. The molecular orbital calculation revealed that the electrons of the HOMO are localized at the triphenylamine group in the TPA. 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https://openalex.org/W3092891643	https://www.matec-conferences.org/10.1051/matecconf/202032103013/pdf	English	null	Topology optimisation of an aircraft nose-wheel fork for production in Ti6Al4V by the Aeroswift high-speed laser powder bed fusion machine	MATEC web of conferences	2,020	cc-by	3,927	MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium Topology optimisation of an aircraft nose-wheel fork for production in Ti6Al4V by the Aeroswift high-speed laser powder bed fusion machine L F Monaheng1, W B du Preez2, N Kotze3, & M Vermeulen4 1,2Central University of Technology, Free State, Bloemfontein, South Africa 3,4ADC Aeroswift, Wonderboom Airport, Pretoria, South Africa 3,4ADC Aeroswift, Wonderboom Airport, Pretoria, South Africa lmonaheng@cut.ac.za © The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution License 4.0 (http://creativecommons.org/licenses/by/4.0/). Abstract Recent levels of structural integrity of components built in the Aeroswift high-speed laser powder bed fusion machine have led to the decision to produce a structural aircraft component through this technology. The Aeroswift machine is capable of building larger Ti6Al4V parts at a more rapid pace than current commercial laser metal powder bed fusion systems. As prototype component, the nose-wheel fork of the AHRLAC aircraft, which was conventionally machined in aluminum alloy 7050, was selected. This paper describes the design, topology optimisation and the manufacturing approach taken in this project. Given the design space, loads, strength requirements and boundary conditions prescribed by the AHRLAC engineers, topology optimisation was performed on the nose-wheel fork to design a lightweight component for production in Ti6Al4V. Different topology optimisation software suites were used, to establish their capabilities and fit-for-purpose features. The optimised design and percentage of weight saving are presented. An assessment based on the experience with the different software suites is offered. Keywords: Topology optimisation, laser metal powder bed fusion, Titanium, nose-wheel fork. https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium 1. Introduction AHRLAC (Advanced High Performance Reconnaissance Light Aircraft) is an unusual two-person cockpit pusher propeller plane, designed by engineers of the South African company Aerosud and manufactured in partnership between the Paramount Group and Aerosud. The AHRLAC company considers additive manufacturing (AM) as a technology fitting well in their manufacturing strategy [1] [2]. The Aeroswift high-speed laser powder bed fusion machine was designed to manufacture metallic parts from alloy powders, such as titanium and stainless steel [1] [3]. This system’s large build volume of 2 m x 0.6 m x 0.6 m allows production of large parts, as well as large batches of small parts. Parts can be produced up to ten times faster than current commercial AM systems [1]. Recent levels of structural integrity of components built in the Aeroswift high-speed laser powder bed fusion machine have led to the decision to produce a structural aircraft component through this technology. As prototype component, the nose-wheel fork of the AHRLAC aircraft, which was conventionally machined in aluminum alloy 7050, was selected. Given the design space, loads, strength requirements and boundary conditions prescribed by the AHRLAC engineers, topology optimisation was performed on the nose-wheel fork to redesign a lightweight nose-wheel fork for production in Ti6Al4V. Different topology optimisation software suites were used to establish their capabilities and fit-for-purpose features. Topology optimisation concentrates on the distribution of the material and structural connectivity in the design domain (space) [4]. The optimisation problem consists of the determination of the optimal distribution of a material in the design domain using a user defined objective function under the given constrains. Two main methods of topology optimisation are the gradient based and non-gradient based methods. In the non-gradient based method material is gradually removed from the structure using a sensitivity function to evolve towards an optimum [5]. For the gradient-based method a mathematical 2 2 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium model (material-indicator function) is derived to calculate the sensitivities of the design variation [6]. The gradient-based optimisation method is used by the structural analysis software solvers MSC Nastran® and Altair OptiStruct®. The gradient-based method Solid Isotropic with Material Penalisation (SIMP), which makes it easy to be implemented in the finite element analysis (FEA) framework, is often used [5]. The SIMP method implements the principle of a panelising density variable to converge to zero (void) or one (solid). 1. Introduction In practice, low-density elements experience high levels of stress and are not removed from the design domain [7]. In this way the optimal shape (load path) is generated from the optimisation software. The topology optimisation process has been applied successfully to minimise weight of aircraft parts. The applications include the design of inboard and outer fixed leading edge ribs, the fuselage door intercostal of the Airbus A380 aircraft, wing box rib and internal structure, spar structures under aeroelastic loads and aircraft structures flutter suppression [8] [9] [10]. It has been demonstrated that the use of topology optimisation for minimising the weight of aircraft parts reduce the fuel consumption and emission gasses, such as carbon dioxide and nitrogen oxide [11]. The combination of topology optimisation with AM of Ti6Al4V (ELI) could advance saving production cost and ensuring positive production in aviation industries [12]. However, this intriguing use of metal AM for production of aircraft structural parts presents great challenges to the designer. Unlike, the conventional method of producing aircraft structural parts, topology optimisation design for additive manufacturing (DfAM) practices are not sufficiently understood nor characterised for production of aircraft structural parts [13]. Therefore, there is a need to establish topology optimisation DfAM as a design tool for manufacturing of aircraft structural parts, with predictable structural performance using conventional design tools. This paper evaluated the capability and fit-for-purpose feature of the topology optimisation software used for the redesign of the AHRLAC nose-wheel fork. 2.2. Topology optimisation with Altair SolidThinking Inspire® MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium A computer-aided design (CAD) domain, derived from the CAD model of the machined nose-wheel fork shown in Figure 1(a) and (c) was created. The volume of the domain was made larger than the machined fork design to ensure design freedom. The bushes were separated from the design domain using the Inspire® partition tool. Additionally, the top contour and dimensions on the design domain, as well as the dimensions of the wheel, shock absorber and torque arm bushes, were kept the same as in the machined fork design, to ensure zero interference between the end-product and other parts of the aircraft. Figure 1(b) and (d) show the design domain with bushes. All loads and boundary conditions were set on the bushes. 2.1. Material properties for AHRLAC nose-wheel fork The nose-wheel fork of the AHRLAC, which was conventionally machined from Al 7050 and is 8,23 kg in weight, was redesigned using topology optimisation software. The redesigned component would be manufactured in the Aeroswift machine from Ti6Al4V (ELI). In Table 1 the material properties used in the design process, as well as the properties of Al 7050 from which the existing nose-wheel fork is produced, are shown. Materials Elastic Modulus (GPa) Poisson’s ratio Density (kg/m3) Yield Strength (MPa) Ti6Al4V (ELI) 110 0.31 4429 787 Al 7050 71 0.33 2830 450 Table 1: Material properties used in the topology optimisation design. 2.2. Topology optimisation with Altair SolidThinking Inspire® Table 1: Material properties used in the topology optimisation design. Table 1: Material properties used in the topology optimisation design. 2.2. Topology optimisation with Altair SolidThinking Inspire® 3 3 https://doi.org/10.1051/matecconf/202032103013 esign domain with bushes derived from the machined fork CAD model. In Figure 2 the two main load configurations of the AHRLAC nose-wheel fork are illustrated. A total of 16 loads cases were provided by the AHRLAC engineers. Four of them were envelope calculated, while twelve conformed to Federal Aviation Regulations. For the maximum landing load case (Figure 2(a)) the fork experiences the loads through the wheel axle, while for the ground static case in Figure 2(b), the load acts on the fork through the tyre (Fz and Fy at the tyre radius). The ground static and maximum load cases are the highest loads experienced by the fork. Therefore, the optimisation was mainly done for these loads. 4 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium Figure 2: Configuration of the maximum landing (a) and ground static (b) load cases calculated by the AHRLAC engineers. Figure 2: Configuration of the maximum landing (a) and ground static (b) load cases calculated by the AHRLAC engineers. 2.3. Topology optimisation with MSC Patran® Optimisation in the MSC Patran® 2018 software, which uses Nastran® 2018.1 as the solver, was based on the same design domain shown in Figure 1(b) and (d). For this process, the design domain was created using SolidWorks® because it is easier to use than Patran® for drawing a complex geometry. The design domain was first imported in the MSC Apex® software for hexahedral mesh generation. A mesh with global element length of 6 mm was generated and its quality was evaluated for invalid elements. The Ti6Al4V (ELI) material properties in Table 1 were set on the bush and design domain generated mesh before the database file (bdf) was exported and imported into the Patran® software. Beam elements were used to represent the wheel, shock absorber, and torque arm axles. At each end of the beam rigid body elements 3 (RBE 3) were used to transfer loads from the beam to the fork hex mesh CAD mode (or design domain with bush) as illustrated in Figure 3. Boundary conditions and loads were applied in the same way as was done in the Inspire® software. Finally, minimising the mass an objective function of a mass fraction of 0.3 was applied. Figure 4 illustrates the load, boundary conditions and mesh for the MSC Patran®-Nastran® optimisation. Figure 4: The load cases, boundary conditions, and mesh generated in the Patran® software, equivalent to that used with the Inspire® software. Figure 4: The load cases, boundary conditions, and mesh generated in the Patran® software, equivalent to that used with the Inspire® software. MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium https://doi.org/10.1051/matecconf/202032103013 The Ti6Al4V (ELI) properties given in Table 1 were assigned on the bushes and design domain. Finally, the objective function was set as maximum stiffness mass target of 30% of the total design domain volume. Figure 2: Configuration of the maximum landing (a) and ground static (b) load cases calculated by the AHRLAC engineers. The mid node of the torque arm axle was restricted to translate only in the Z direction. On the mid-point of the shock absorber axle, only rotation in the Y axis was allowed. Finally, the shape control, symmetric about the Z-X plane as illustrated in Figure 3, was set. Figure 3: The connectors, all load cases, symmetric plane, and boundary conditions as set in the fork for topology optimisation. Figure 3: The connectors, all load cases, symmetric plane, and boundary conditions as set in the fork for topology optimisation. 5 5 Figure 6: Views of the final design. (a) resulted from Altair SolidThinking Inspire® and (b) from the MSC Patran®- Nastran®. Figure 6: Views of the final design. (a) resulted from Altair SolidThinking Inspire® and (b) from the MSC Patran®- Nastran®. Figure 6: Views of the final design. (a) resulted from Altair SolidThinking Inspire® and (b) from the MSC Patran®- Nastran®. The smooth optimal nose-wheel fork CAD model created from the Inspire® software was imported into the Patran® software for FEA purpose. A tetrahedral solid mesh was generated on the model with global element size of 10 mm. The beam element, rigid body element 3, Ti6Al4V (ELI) properties together with the loads and boundary conditions were set on the final design CAD model in the same way as was done in the MSC topology optimisation. The Von Mises stress and maximum displacement experienced by the fork was computed for the maximum landing and ground static load cases. In Figure 7(a) and (b) the maximum Von Mises stress and the corresponding displacement are shown. Similarly, Figure 7(c) and (d) illustrate the stress experienced during the ground static load case with its respective displacement. Figure 7: FEA results (a) and (b) for maximum load case and (c) and (d) for ground static load case. Figure 7: FEA results (a) and (b) for maximum load case and (c) and (d) for ground static load case. The Von Mises stress calculated for the maximum and ground static load cases are 505 MPa and 426 MPa, respectively The maximum displacement for these same load cases are 13.2 mm and 12.2 mm, respectively. The Von Mises stress calculated for the maximum and ground static load cases are 505 MPa and 426 MPa, respectively. The maximum displacement for these same load cases are 13.2 mm and 12.2 mm, respectively. 3. Results In Figure 5(a) and (b) the Altair SolidThinking Inspire® and MSC Patran®-Nastran® topology optimisation results, respectively, are presented. The different views of the results are shown to demonstrate the similarity and the differences between the Altair and MSC topology optimisation software. These results illustrate the load path or the elements which experience more load. The top and bottom boundary elements carry more load and, therefore, were not removed from the design domain. On the other hand, the elements which experience low levels of stress were removed from the design space as seen for both the Altair and MSC optimisation results. 6 6 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium Figure 5: Views of the topology optimisation results. (a) Results from Inspire® software and the (b) from the Patran®-Nastran® software. Figure 5: Views of the topology optimisation results. (a) Results from Inspire® software and the (b) from the Patran®-Nastran® software. Once the load path was determined, a smooth CAD model was generated to create the final design. The PolyNURB feature of the Altair SolidThinking Inspire® software was used to trace over the optimisation results with precision to create a free-form smooth geometry CAD model shown in Figure 6(a). The final design had a mass of 6.62 kg. which is a 20% mass saving as compared to the machined fork design. Finally, the model was exported as STEP file for FEA. In MSC Patran® the creation of geometry from elements is not automated, although not impossible, and the elements smoothing feature does not provide good quality, see Figure 6(b). 7 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium 4.1. Assessment of the topology optimisation software fit-for-purpose feature 4.1. Assessment of the topology optimisation software fit-for-purpose feature Topology optimisation is possible with both Inspire® and Patran® software. At the no-design portion (bushes) no elements were removed to avoid introducing voids in the CAD model. However, elements were removed at portions of the design domain where the stresses were calculated to be low using FEA. The region of the design domain which experienced higher stress was kept solid. The connectivity of the solid portion in the design domain provide the load path. In this way weight is reduced from the design space by the topology optimisation software. Different topology optimisation software suites from different vendors have different end user icons and optimisation approaches. In Table 2 the requirements of optimisation for the design of the landing gear fork are summarised for both software suites. 8 8 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium Table 2: The requirements of topology optimisation for the design of the nose-wheel fork. Topology optimisation requirements Altair SolidThinking Inspire® MSC Patran® 1. CAD model Large design domain is necessary. Creating it from the existing model is quick and fast using Inspire® geometry tools such as push-pull, simplifier, partition. The design space creation from the existing CAD model could be time consuming for a complex geometry such as the landing gear fork. It was best done in SolidWorks®. 2. Mesh Mesh generation is not user based. It is robust and automatically applied on the CAD model. For accurate results fine hexahedral mesh is necessary. Generation of such mesh requires decomposition of complex CAD model in cuboids and/or primitive shape, which can be tedious, complex and difficult to automate. MSC Apex® software was used for creation of hexahedral mesh. 3. Material properties In the material library Ti6Al4V (ELI) is available. Additionally, a new material can be added in the library. Material properties must be added in. 4. Loads & boundary conditions Load and boundary conditions are auto detected. For example, when applied on the cylindrical face of the model, bearing load is automatically suggested. However, it is still up to the user to define the desired loads and supports. Finally, constraints are applied on the non-design part of the model. The definition of the loads and supports are required from the user. 5. Solver Optistruct® software is integrated in Inspire® as the solver. 4.1. Assessment of the topology optimisation software fit-for-purpose feature It alters material distribution to optimise the user defined objective function under given constraints using the gradient based optimisation method. Nastran® software is based on the SIMP method for topology optimisation in which a power-law is used as penalty function. Table 2: The requirements of topology optimisation for the design of the nose-wheel fork. For accurate results fine hexahedral mesh is necessary. Generation of such mesh requires decomposition of complex CAD model in cuboids and/or primitive shape, which can be tedious, complex and difficult to automate. MSC Apex® software was used for creation of hexahedral mesh. Nastran® software is based on the SIMP method for topology optimisation in which a power-law is used as penalty function. 9 9 https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium 6. Topology optimisation post-process Topology optimisation post-process refers to conversion of the mesh, as shown in Figure 5, which is rough at portions where elements with less stress were removed, into a smooth solid CAD model. With the PolyNURB® function available in Inspire® software complex shapes, which best represent the optimisation results can be achieved. Using Inspire® in conjunction with CAD software such as SolidWorks® a best design of the nose-wheel fork can be repeatedly obtained as seen in Figure 6(a). FEA is an essential post topology optimisation to verify if the design can handle all the loads. The conversion of elements into CAD model, is not automated and the mesh smoothing quality not good as illustrated in Figure 5(b). However, the model can be redrawn to resemble the optimisation results illustrated. For this complex geometry the process is tedious, and mismatch is highly possible. Several FEA iterations will be necessary to verify that the design of the nosewheel fork can handle all the loads. 7. Data Storage Single file was generated during topology optimisation. The size of the file is 5.5 Mb. Besides the database file, multiple files, such as journal, master, session, were generated. The total file size amounts to 5.5 Gb. 4.2. Success of the topology optimisation design for additive manufacturing The conversion of elements into CAD model, is not automated and the mesh smoothing quality not good as illustrated in Figure 5(b). However, the model can be redrawn to resemble the optimisation results illustrated. For this complex geometry the process is tedious, and mismatch is highly possible. 5. Conclusions Topology optimisation design of the nose-wheel fork for production in Ti6Al4V (ELI) using the Aeroswift machine is possible with both the Altair SolidThinking Inspire® and MSC Patran®-Nastran® software. However, the user features and processes are different. Inspire® has more user-friendly features than the MCS Patran®-Nastran®software, making it the better and more time-effective tool for topology optimisation. 4.1. Assessment of the topology optimisation software fit-for-purpose feature Several FEA iterations will be necessary to verify that the design of the nosewheel fork can handle all the loads. 6. Topology optimisation post-process Topology optimisation post-process refers to conversion of the mesh, as shown in Figure 5, which is rough at portions where elements with less stress were removed, into a smooth solid CAD model. With the PolyNURB® function available in Inspire® software complex shapes, which best represent the optimisation results can be achieved. Using Inspire® in conjunction with CAD software such as SolidWorks® a best design of the nose-wheel fork can be repeatedly obtained as seen in Figure 6(a). Besides the database file, multiple files, such as journal, master, session, were generated. The total file size amounts to 5.5 Gb. 4.2. Success of the topology optimisation design for additive manufacturing 4.2. Success of the topology optimisation design for additive manufacturing The yield strength of parts built in the Aeroswift machine from Ti6Al4V is 787 MPa. The maximum Von Mises stress computed in both the maximum landing and ground static load cases is less that this yield strength. The total safety factor calculated was 1.6. The optimised final design was 20% less in weight than the machined fork design. 6. Acknowledgements This research was supported and funded by the South African Department of Science and Technology through the Collaborative Program in Additive Manufacturing, Contract No.: CSIR-NLC-CPAM-15-MOA-CUT-01. 10 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium https://doi.org/10.1051/matecconf/202032103013 MATEC Web of Conferences 321, 03013 (2020) The 14th World Conference on Titanium MATEC Web of Conferences 321, 03013 (2020) https://doi.org/10.1051/matecconf/202032103013 The 14th World Conference on Titanium 7. References [1] P. Middleton, African Fusion, Aeroswift as large-scale powder bed fusion AM system, Crown Publications. (2018) 20–21. [2] D. de Beer, W. du Preez, H. Greyling, F. Prinsloo, F. Sciammarella, N. Trollip, M. Vermeulen, with contributions from Terry Wohlers, A South African Additive Manufacturing Strategy, Department of Science and Technology, South African Additive Manufacturing Strategy. (2016). [3] B. P. Conner, G.P. 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https://openalex.org/W4316813095	https://www.researchsquare.com/article/rs-2234261/latest.pdf	English	null	Deep learning-based prediction of mandibular growth trend in children with anterior crossbite using cephalometric radiographs	BMC oral health	2,023	cc-by	5,477	Deep learning-based prediction of mandibular growth trend in children with anterior crossbite using cephalometric radiographs Jia-Nan Zhang Sir Run Run Shaw Hospital Hai-Ping Lu Zhejiang Chinese Medical University Jia Hou Hangzhou Dianzi University Qiong Wang Sir Run Run Shaw Hospital Feng-Yang Yu Perfect Dental Care Chong Zhong Perfect Dental Care Cheng-Yi Huang Sir Run Run Shaw Hospital Si Chen (  elisa02@163.com ) Peking University School and Hospita Conclusions The deep-learning CNN model could predict the growth trend of the mandible in anterior crossbite children with relatively high accuracy using cephalometric images. The prediction decision was made by a direct and comprehensive detecting and analyzing system instead of doctor’s opinion from clinical experience. Results The deep-learning model showed a good prediction accuracy about 85%, much higher when compared with the 54.2% of the junior orthodontists. The sensitivity and specificity of the model was 0.95 and 0.75 respectively, higher than that of the junior orthodontists (0.62 and 0.47 respectively). The area under the curve (AUC) value of the deep-learning model was 0.9775. Visual inspection showed that the model mainly focused on the characteristics of special regions including chin, lower edge of the mandible, incisor teeth, airway and condyle to conduct the prediction. Background It is difficult for orthodontists to accurately predict the growth trend of the mandible in children with anterior crossbite. This study aims to develop a deep learning model to automatically predict the mandibular growth result into normal or overdeveloped using cephalometric radiographs. Methods A deep convolutional neural network (CNN) model was constructed based on the algorithm ResNet50 and trained on the basis of 256 cephalometric radiographs. The prediction behavior of the model was tested on 40 cephalograms and visualized by equipped with Grad-CAM. The prediction performance of the CNN model was compared with that of three junior orthodontists. Research Article Posted Date: November 15th, 2022 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Additional Declarations: No competing interests reported. Page 1/16 Version of Record: A version of this preprint was published at BMC Oral Health on January 17th, 2023. See the published version at https://doi.org/10.1186/s12903-023-02734-4. Version of Record: A version of this preprint was published at BMC Oral Health on January 17th, 2023. See the published version at https://doi.org/10.1186/s12903-023-02734-4. Version of Record: A version of this preprint was published at BMC Oral Health on January 17th, 2023. See the published version at https://doi.org/10.1186/s12903-023-02734-4. Page 2/16 Background Class III malocclusion is a frequently observed clinical problem, mostly manifested as anterior crossbite, occurring in 4%-14% East Asian populations [1]. Anterior crossbite not only affects the occlusal function, but also damages the balance of profile, and increases the social and psychological burden of the child [2]. As for the components of Class III malocclusion, the combination of underdeveloped maxilla and overdeveloped mandible was most common at 1/3, whereas those with a normal maxilla and overdeveloped mandible constituted about 1/5 [3]. Children who have Class III malocclusion because of excessive growth of the mandible are extremely difficult to treat. Prediction for the future growth trend of Page 3/16 Page 3/16 the mandible greatly affects the decision for treatment option. Inaccurate prediction may lead to insufficient or unnecessary treatment. In fact, it is difficult for orthodontists to accurately predict the remaining growth amount of the mandible. Similar anterior crossbite in the mixed dentition may develop to malocclusions with different skeletal pattern in the permanent dentition [4]. Therefore, accurate prediction of the future mandibular growth in the early stage will be very useful to assist treatment planning and prognosis. Various mathematical methods have been proposed over the past few decades to predict the growth of mandible. Sato [5] established a linear equation to predict mandibular growth potential based on bone age, which was assessed from hand-wrist radiographs. This method was not widely used due to the unnecessary radiation dose to children resulted from the additional hand-wrist radiograph. Later, Mito [6] developed a formula to predict mandibular growth potential by calculating the actual growth of the mandible (condyle-gnathion) with cervical vertebral bone age, which was evaluated from cephalometric radiographs. However, this method was not suitable for individual prediction because the formula was derived from the mixed data of 7–13 years of age children, including different stages of the growth period. Moshfeghi [7] set a regression equation to predict mandibular length (Articulare-Pogonion) by analyzing the morphological changes of the cervical vertebrae on lateral cephalograms. Recently, Franchi [8] developed a mathematical mixed effect model to predict the growth of mandible, and the results demonstrated that cervical stage, chronological age and gender were significant predictors for the annualized increments in mandibular growth. Among the above studies, regression equation analysis is the most frequently used method for predicting mandibular growth potential. Patients Inclusion This retrospective study was approved by the Research Ethics Committee of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine (No.20210729-122). In this study, 512 patients who visited our Orthodontic Center between January 2010 and December 2016 with the chief complain of anterior crossbite were screened for further research. The inclusion criteria were as follows: (1) anterior crossbite; (2) Class III or Class I molar relationship; (3) ANB < 0°; (4) without functional mandibular setback to edge to edge; (5) 8–14 years of age; (6) availability of the pre-treatment (T1) and after 18-year of age (T2) lateral cephalograms which were of good quality. The exclusion criteria were as follows: (1) maxillary retrusion; (2) anterior crossbite caused by misaligned teeth; (3) congenital deformity such as cleft lip and palate, infection or trauma history. A total of 296 patients were included in this study (142 males and 154 females, ranged from 8.08 to 13.92 years, with an average age of 10.8 years). Background Regression equation is applied to identify and verify the factors that probably reflect the growth potential of mandible [9]. However, as it works based on the analysis of a linear combination of covariates, it may be too simplistic to predict the complex growth outcome of mandible. Deep learning technique is a big breakthrough in machine learning field, and it has showed great application potential in medical filed in recent years. Deep learning models have behaved high accuracy on medical image classification by automatically learning from datasets, which are manually annotated by clinical experts [10, 11]. Among deep learning models, the convolutional neural network (CNN) achieves the most attention and is widely researched due to its amazing performance for detection of medical images [12]. CNN models can automatically learn and extract characteristic features and structures from training images, and then make classification and prediction for new images. In the medical field, detection and classification of skin cancer and pulmonary tuberculosis by CNN models have already been verified to be of high accuracy and efficiency [13, 14]. In the dental field, Kim [15] used a deep CNN model to automatically identify and classify skeletal malocclusions into three classes from 3D Cone-Beam Computerized Tomography (CBCT) craniofacial images and achieved a good performance of the accuracy over 93%. Yu [16] constructed a multimodal CNN model to provide vertical and sagittal skeletal diagnosis with cephalometric radiographs and reported a high classification Page 4/16 accuracy at 96.4%. The CNN model has showed its great potential for the analysis of Orthodontic X-ray images, including CBCT and cephalograms. However, to our knowledge, prediction for the growth potential of the mandible based on CNN models has not yet been reported. accuracy at 96.4%. The CNN model has showed its great potential for the analysis of Orthodontic X-ray images, including CBCT and cephalograms. However, to our knowledge, prediction for the growth potential of the mandible based on CNN models has not yet been reported. In this research, a deep learning CNN model was trained to automatically classify the mandibular growth result for children with anterior crossbite into two types: normal and overdeveloped using cephalometric radiographs. The analysis behavior and process of the CNN model could be visualized by highlighting a heatmap on the cephalometric image. The prediction results from the CNN model were compared with those from the junior orthodontists to explore its clinical application. Architecture Of The Deep-learning Model We developed a neural network based on ResNet50, which was famous for its excellent performance in image classification and object detection [18]. The architecture of this model was composed of several residual networks and a softmax layer. The residual network was used to detect and analyze the characteristics of the input images. The softmax layer was adopted to predict the classification of the object. Figure 1 showed the workflow of this deep learning-based CNN model. The training process was performed on a Linux machine with a GPU accelerator, and the initial learning rate and training epoch was 0.01 and 300, respectively. Cephalometric Analysis And Skeletal Classification All T1 and T2 cephalometric radiographs were uploaded into Dolphin software (Version 11.9, Dolphin Digital Imaging, Chatsworth, Calif, USA). The anatomic contours were traced and cephalometric landmarks were located simultaneously by two orthodontic experts. Any disagreements about landmark location were resolved by retracing the anatomic contours until the two experts achieved the same point. The cephalometric measurements related to evaluate the growth condition of mandible included SNB, ANB, Wits appraisal, FMA (mandibular plane to FH), SNPog (facial plane to SN), NSGn (Y-Axis), NSAr (Sella Angle), ArGoMe (Gonial Angle); Ar-Gn (effective mandibular length), Co-Gn (total mandibular length), Go-Gn (mandibular body length), and Co-Go (mandibular ramus length). Page 5/16 Page 5/16 Page 5/16 Anterior crossbite with prognathic mandible belongs to skeletal malocclusion which may require orthognathic surgery according to the Kerr’s research [17]. In the contrast, anterior crossbite with normal mandible can be treated by orthodontics. According to the cephalometric analysis results at T2, if SNB > 86°, ANB < -2° and Wits value < -2.0mm [17], the subject was recognized as a patient with overdeveloped mandible and assigned to Group A, otherwise, assigned to Group B (patient with normal mandible). Finally, 102 patients (49 males and 53 females, ranged from 8.08 to 13.92 years, with an average age of 11.5 years) were sorted to Group A and 194 patients (93 males and 101 females, ranged from 8.08 to 13.83 years, with an average age of 10.4 years) were sorted to Group B. Anterior crossbite with prognathic mandible belongs to skeletal malocclusion which may require orthognathic surgery according to the Kerr’s research [17]. In the contrast, anterior crossbite with normal mandible can be treated by orthodontics. According to the cephalometric analysis results at T2, if SNB > 86°, ANB < -2° and Wits value < -2.0mm [17], the subject was recognized as a patient with overdeveloped mandible and assigned to Group A, otherwise, assigned to Group B (patient with normal mandible). Finally, 102 patients (49 males and 53 females, ranged from 8.08 to 13.92 years, with an average age of 11.5 years) were sorted to Group A and 194 patients (93 males and 101 females, ranged from 8.08 to 13.83 years, with an average age of 10.4 years) were sorted to Group B. Datasets Build And Annotation The lateral cephalometric images of the 296 subjects at T1 were collected for the training and testing of the deep learning-based CNN model. Among those, 256 lateral cephalograms (82 images from Group A and 174 images from Group B) were randomly selected as the training dataset. The remaining 40 cephalometric images (20 images from Group A and 20 images from Group B) were used as the testing dataset to evaluate the performance of the deep learning-based CNN model. The input cephalometric images were cropped and resized to 512*512 pixels without changing its aspect ratio, aiming to reduce redundant information and improve the efficiency of the training process. Then, in order to avoid overfitting, the images were randomly augmented by applying random transformations, including rotation, horizontal and vertical flipping, width and height shifting, shearing, and zooming. Based on the classification results of Group A and Group B, the reference annotations of the mandibular growth trend (overdeveloped vs normal) for T1 cephalograms were created. Visualization Of Region Of Interest The classification behavior of the model was recorded and visualized by marking a region of interest (ROI) on the input image, using a visualization method called “Grad-CAM”. Grad-CAM is a region proposal network that is equipped into the output layer of the neural network and can mark the ROI [19]. Page 6/16 Page 6/16 Specifically, Grad-CAM has the super advantage of localizing the most discriminative and critical region from the whole scene for classifying the input image because some special spatial element in the feature maps plays an essential role in the calculating and prediction process of the model. Mandibular Growth Prediction And Statistical Analysis After completion of the training process of the CNN model, the testing dataset (20 images from Group A and 20 images from Group B) was classified by the CNN model and junior orthodontists respectively. For CNN model classification, the testing images were input to the model and the classification result will be given based on the possibility comparison between different classification. For example, as shown in Fig. 2, after the input of Image X, the output result was shown as: 0.9963781 for Class A (overdeveloped mandible) and 0.0036219 for Class B (normal mandible). Then Image X was classified to Class A. For junior orthodontist classification, three junior orthodontists (clinical work experience less than five years) gave their individual judgement for the mandibular growth of the subject based on the testing cephalogram only. The performance of the deep learning-based CNN model and the junior orthodontists were compared by the following indices: classification accuracy, true positive rate (sensitivity), false negative rate, false positive rate, true negative rate (specificity), and the area under the curve (AUC). These calculating work were based on a Keras framework in Python. Comparison of Prediction Results The performance of the deep-learning CNN model and the junior orthodontists were summarized in Table 1. Higher accuracy was found in the deep-learning model prediction results (85.0%) when compared with that of the three junior orthodontists (54.2%). The mean sensitivity/specificity for the model prediction and human prediction were 0.95/0.75 and 0.62/0.47 respectively. The receiver-operating characteristics (ROC) curve of the model and the performance points of the junior orthodontists in the ROC picture were shown in Fig. 3. The AUC value of the deep-learning model was 0.9775. Page 7/16 Table 1 Performance of the deep learning model and the junior orthodontists. Accuracy TPR FNR FPR TNR AUC The Deep Learning Model 85% 0.95 0.05 0.25 0.75 0.9775 The Junior orthodontists (Mean Value) 54.2% 0.62 0.38 0.53 0.47 / Orthodontist 1 57.5% 0.8 0.2 0.65 0.35 / Orthodontist 2 50% 0.45 0.55 0.45 0.55 / Orthodontist 3 55% 0.6 0.4 0.5 0.5 / TPR, true positive rate (sensitivity); FNR, false negative rate; FPR, false positive rate; TNR, true negative rate (specificity); AUC, area under the curve. Table 1 TPR, true positive rate (sensitivity); FNR, false negative rate; FPR, false positive rate; TNR, true negative rate (specificity); AUC, area under the curve. Visualization Of Localization Results Figure 4 showed some examples of the heatmap images reflecting the characteristic of the learning and classification behaviors of the deep-learning model. Visual inspection results showed that the deep- learning model mainly focused on the following areas: chin (40/40), lower edge of the mandible (28/40) and incisor teeth (7/40). It’s interesting to find that the area used to be considered as not important for the prediction of the mandibular growth such as the airway area was recognized as ROI in some subjects by the deep-learning model (2/40) (Table 2). Table 2 The occurrence times of the regions of interest among the heatmaps of the 40 testing images. Chin lower edge of the mandible incisor teeth airway condyle The Occurrence Times 40 28 7 2 2 Discussion This deep learning model was developed from a well-known algorithm called ResNet, which has been recognized as a breakthrough innovation for its strong ability to train extremely deep neural networks, possessing important advantages such as analyzing more structure information, relevance information and detail information of images [25]. In the deep learning field, the AUC value is considered to be an important index when evaluating the performance of the models [26]. The deep learning model in this study achieved an AUC value of 0.9775, demonstrating its reliable performance. In the past few years, deep learning techniques have been successfully applied to the dental field and acquired significant achievements. Deep learning models have showed a reliable ability in identifying and classifying several types of dental imaging, even surpassing human experts [22, 23]. In this study, a deep learning-based CNN model was trained to predict the growth trend of mandible in anterior crossbite child from pre-treatment cephalometric radiographs. The result showed that the deep learning model achieved 85% accuracy in predicting whether the mandible of anterior crossbite child will grow into an overdeveloped mandible or a normal mandible. The accuracy of the machine learning based prediction is much higher than that from the junior orthodontists, which was 54.2%. The sensitivity and specificity for the model prediction were also higher when compared with human prediction (0.95 vs 0.62 and 0.75 vs 0.47). The reason for the good performance of this deep learning model should be related to the innovation of the prediction process, which replaced the analysis of linear and angular measurements by a direct and comprehensive detecting and analyzing system [24]. This deep learning model was developed from a well-known algorithm called ResNet, which has been recognized as a breakthrough innovation for its strong ability to train extremely deep neural networks, possessing important advantages such as analyzing more structure information, relevance information and detail information of images [25]. In the deep learning field, the AUC value is considered to be an important index when evaluating the performance of the models [26]. The deep learning model in this study achieved an AUC value of 0.9775, demonstrating its reliable performance. Among the results from this study, there were two that worth more notice. One was that the false positive rate of the deep learning model was much lower than that of the junior orthodontists. Discussion Class III malocclusion can be of dental or skeletal origin, so it is crucial to classify the malocclusion accurately in order to manage it on a sound clinical basis. Class III malocclusion resulted from overdeveloped mandible is extremely difficult to treat. The mandible has the longest growth and develop period among the craniofacial bones [20]. Therefore, the prediction for the mandibular growth in the early stage is very challenging. Severe skeletal Class III with prognathic mandible requires orthognathic surgery. However, if camouflaged orthodontic extraction treatment was conducted at an early stage, the result might be unstable or even deteriorated during the treatment. Since the key point of successful treatment of anterior crossbite lies on the mandible, the early and accurate prediction of the growth trend of mandible and treatment with corresponding interventional Page 8/16 Page 8/16 methods can be beneficial to the patients [21]. Clinically, the decision of proper treatment and timing for Class III patients mainly relies on the clinical experience of orthodontists. However, individual clinical experience varies significantly among different orthodontists. On the other hand, even for experienced orthodontists, it is also challenging to accurately predict the remaining growth of mandible when facing anterior crossbite in mixed dentition or early permanent dentition. In the past few years, deep learning techniques have been successfully applied to the dental field and acquired significant achievements. Deep learning models have showed a reliable ability in identifying and classifying several types of dental imaging, even surpassing human experts [22, 23]. In this study, a deep learning-based CNN model was trained to predict the growth trend of mandible in anterior crossbite child from pre-treatment cephalometric radiographs. The result showed that the deep learning model achieved 85% accuracy in predicting whether the mandible of anterior crossbite child will grow into an overdeveloped mandible or a normal mandible. The accuracy of the machine learning based prediction is much higher than that from the junior orthodontists, which was 54.2%. The sensitivity and specificity for the model prediction were also higher when compared with human prediction (0.95 vs 0.62 and 0.75 vs 0.47). The reason for the good performance of this deep learning model should be related to the innovation of the prediction process, which replaced the analysis of linear and angular measurements by a direct and comprehensive detecting and analyzing system [24]. Discussion This suggested that orthodontists with less clinical experience tended to be overcautious in the prediction of future mandibular growth in order to reducing clinical risk and avoiding medical dispute. Another revealing was that the false negative rate of the deep learning model was also much lower than that of the junior orthodontists. This implied that junior orthodontists might be more likely to make a wrong prediction of judging the mandible to be normal in the future, whereas the mandible would grow into an excessive size. This type of wrong prediction may result in unsuccessful camouflaged orthodontic treatment for cases which actually need orthognathic surgery and anterior crossbite relapse after treatment. In addition, we combined Grad-CAM into the deep learning model to provide a guide for visually predicting the growth feature of the mandible in anterior crossbite child and reveal the prediction mechanism behind. The output of this model includes heatmaps, which allowing the identification of the main areas in cephalometric images by which the model made the prediction decision. The results showed that the heatmaps included regions of chin, lower edge of the mandible, incisor teeth area, airway and condyle. The regions like chin and lower edge of the mandible are easy to understand as they are closely related to Page 9/16 Page 9/16 the growth regulation of mandible [27, 28]. However, the airway region is unexpectable as we rarely take it into consideration for mandibular growth prediction in anterior crossbite child. The findings from this study provided some new clues for prediction. Despite the good performance of the deep learning-based CNN model, there are several limitations in our approach. First, the total size of the training dataset was small, as well as the testing dataset. Second, other deep learning models were not applied to compare the prediction performance. Last, clinical characteristics and family history of patients were not included in the algorithm, so it is unknown whether the performance of the deep learning model could be improved if these factors were added. Conclusions 1. The deep learning model behaved well and resulted in a much higher accuracy in mandibular growth trend prediction for children with anterior crossbite. 1. The deep learning model behaved well and resulted in a much higher accuracy in mandibular growth trend prediction for children with anterior crossbite. 2. The deep learning model made the prediction decision mainly by identifying the characteristics of the regions of chin, lower edge of the mandible, incisor teeth area, airway and condyle in cephalometric images. 2. The deep learning model made the prediction decision mainly by identifying the characteristics of the regions of chin, lower edge of the mandible, incisor teeth area, airway and condyle in cephalometric images. Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Consent for publication Not applicable. Ethics approval and consent to participate This retrospective study was conducted according to the guidelines of the Declaration of Helsinki and approved by the Research Ethics Committee of Sir Run Run Shaw Hospital, Zhejiang University School of Medicine (No.20210729-122). All the methods in the study were carried out in accordance with the relevant guidelines and regulations. Informed consent agreement was signed by a parent or guardian for participants. Authors' contributions JNZ: writing original draft, data curation, investigation, methodology. HPL: writing review and editing, validation, formal analysis. JH: software. QW: data curation, investigation, methodology. FYY: investigation, methodology. CZ: investigation, methodology. CYH: data curation, investigation, methodology, resources, supervision. SC: conceptualization, writing review and editing, funding acquisition, project administration. All authors have read and approved the final manuscript. Abbreviations Page 10/16 Abbreviations CNN Convolutional Neural Network CBCT Cone-Beam Computerized Tomography Conv Convolution BN Batch Norm ReLU Rectified Linear Unit ROI Region of Interest AUC Area Under the Curve TPR True Positive Rate FNR False Negative Rate FPR False Positive Rate TNR True Negative Rate ROC Receiver-Operating Characteristics CNN Convolutional Neural Network CNN Convolutional Neural Network Page 10/16 Funding This study was supported by three research projects. One is “National Natural Science Foundation of China (81200806)” which provided financial support in stage of data collection and statistical interpretation of data. Another is “Ningxia Hui Autonomous Region key Research and Development Program (2022BEG02031)” which provided financial support in stage of deep-learning model development. The third is “Medical Science and Technology Project of Zhejiang Province (2023KY122)” which provided financial support in stage of open access publishing. The funders had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript. Competing interests The authors declare that they have no competing interests. Authors' information 1Center of Orthodontics, Department of Dentistry, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 3# Qingchundong Road, Hangzhou, China. 2Department of Orthodontics, College of Stomatology, Zhejiang Chinese Medical University, 548# Binwen Road, Hangzhou, China. 3School of Automation, Hangzhou Dianzi University, 1158# 2nd Street, Hangzhou, China; Lishui Institute, Hangzhou Dianzi University, 1# Xueyuan Road, Lishui, China. 4Center of Orthodontics, Perfect Dental Care, 108# Xintang Road, Hangzhou, China. 5Department of Orthodontics, Peking University School and Hospital of Stomatology, 22# Zhongguancun S. Ave., Beijing, China. Acknowledgements Not applicable. Page 11/16 References 1. Ngan P, Moon W. 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A deep learning algorithm for detection of oral cavity squamous cell carcinoma from photographic images: A retrospective study. EClinicalMedicine. 2020;27:100558. 24. Chan HP, Samala RK, Hadjiiski LM, Zhou C. Deep Learning in Medical Image Analysis. Adv Exp Med Biol. 2020;1213:3–21. 25. Yu X, Kang C, Guttery DS, Kadry S, Chen Y, Zhang YD. ResNet-SCDA-50 for Breast Abnormality Classification. IEEE/ACM Trans Comput Biol Bioinform. 2021;18(1):94–102. 25. Yu X, Kang C, Guttery DS, Kadry S, Chen Y, Zhang YD. ResNet-SCDA-50 for Breast Abnormality Classification. IEEE/ACM Trans Comput Biol Bioinform. 2021;18(1):94–102. 26. Poplin R, Varadarajan AV, Blumer K, Liu Y, McConnell MV, Corrado GS, Peng L, Webster DR. Prediction of cardiovascular risk factors from retinal fundus photographs via deep learning. Nat Biomed Eng. 2018;2(3):158–64. Page 13/16 27. Buschang PH, Gandini Júnior LG. Mandibular skeletal growth and modelling between 10 and 15 years of age. Eur J Orthod. 2002;24(1):69–79. 27. Buschang PH, Gandini Júnior LG. The work flow and architecture of the deep learning model. Conv: Convolution. BN: Batch Norm. ReLU: Rectified Linear Unit. Figures Figure 1 The work flow and architecture of the deep learning model. Conv: Convolution. BN: Batch Norm. ReLU: Rectified Linear Unit. Figure 1 Th k fl d hi f h d l i d l C C l i BN B h N R L Figure 1 Figure 1 The work flow and architecture of the deep learning model. Conv: Convolution. BN: Batch Norm. ReLU: Rectified Linear Unit. The work flow and architecture of the deep learning model. Conv: Convolution. BN: Batch Norm. ReLU: Rectified Linear Unit. Page 14/16 Figure 2 The example of the classification process by the deep-learning CNN model. Class A: grow to overdeveloped mandible. Class B: grow to normal mandible. The classification result was verified by the real growth result according to the lateral cephalogram after 18-year of age. Page 15/16 Figure 3 Figure 3 Figure 3 Page 15/16 Receiver operating characteristic (ROC) curve of the deep learning model and the performance points of the junior orthodontists. The solid bule line displayed the trajectory of the deep learning model with respect to Sensitivity (True Positive Rate) and 1-Specificity (False Positive Rate). The colorful points represented the performance of the junior orthodontists. None of the junior orthodontists outperformed the deep learning model. Figure 4 Examples of Grad-CAM localizing ROIs in the cephalometric radiographs. Red colors indicated areas of high influence on the classification making, whereas bule colors indicated areas of low influence. Figure 4 Figure 4 Examples of Grad-CAM localizing ROIs in the cephalometric radiographs. Red colors indicated areas of high influence on the classification making, whereas bule colors indicated areas of low influence. Examples of Grad-CAM localizing ROIs in the cephalometric radiographs. Red colors indicated areas of high influence on the classification making, whereas bule colors indicated areas of low influence. Page 16/16
https://openalex.org/W4229029978	https://www.researchsquare.com/article/rs-1004428/latest.pdf	English	null	Experimental and numerical study on surface generated mechanism of robotic belt grinding process considering the dynamic deformation of elastic contact wheel	The international journal of advanced manufacturing technology/International journal, advanced manufacturing technology	2,022	cc-by	7,841	Experimental And Numerical Study On Surface Generated Mechanism of Robotic Belt Grinding Process Considering The Dynamic Deformation of Elastic Contact Wheel Mingjun Liu Northeastern University Yadong Gong (  ydgong@mail.neu.edu.cn ) Northeastern University Jingyu Sun Northeastern University Yuxin Zhao Northeastern University Yao Sun Northeastern University Northeastern University Research Article Keywords: Surface generated mechanism, Robotic belt grinding, Elastic contact, Dynamic deformation Posted Date: October 26th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-1004428/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at The International Journal of Advanced Manufacturing Technology on April 16th, 2022. See the published version at https://doi.org/10.1007/s00170-022-09067-6. Research Article Keywords: Surface generated mechanism, Robotic belt grinding, Elastic contact, Dynamic deformation Posted Date: October 26th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-1004428/v1 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Posted Date: October 26th, 2021 License:   This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published at The International Journal of Advanced Manufacturing Technology on April 16th, 2022. See the published version at https://doi.org/10.1007/s00170-022-09067-6. [Title Page] Article Title Experimental and numerical study on surface generated mechanism of robotic belt grinding process considering the dynamic deformation of elastic contact wheel Authors Mingjun Liu, Yadong Gong, Jingyu Sun, Yuxin Zhao, Yao Sun Author affiliations: Northeastern University, Shenyang, China Correspondence information: Correspondence author name: Y.D. Gong Affiliation: Northeastern University, Shenyang, China Department address: School of Mechanical Engineering and Automation: Northeastern University, Shenyang, P.R.China 110819 Email address: gongyd@mail.neu.edu.cn Telephone number: 86-139-4051-8488 [Title Page] Abstract In the robotic belt grinding process, the elastic contact condition between the flexible tool and the workpiece is a critical issue which extremely influences the surface quality of the manufactured part. The existing analysis of elastic removal mechanism is based on the statistic contact condition but ignoring the dynamic removal phenomenon. In this paper, we discussed the dynamic contact pressure distribution caused by the non-unique removal depth in the grinding process. Based on the analysis of the equivalent removal depth of a single grit and the trajectories of grits in manufacturing procedure, an elastic grinding surface topography model was established with the consideration of the dynamic contact condition in the removing process. Robotic belt grinding experiments were accomplished to validate the precision of this model, while the result showed that the surface roughness prediction error could be confined to 11.6%, which meant this model provided higher accuracy than the traditional predicting methods. Keywords: Surface generated mechanism; Robotic belt grinding; Elastic contact; Dynamic deformation as, the length of the contact area in the static contact condition. ad, the length of the contact area in the dynamic contact condition. ad, the length of the contact area in the dynamic contact condition. ap,grit(x), the removal depth of a single grit. ap,grit(x), the removal depth of a single grit. , ( ) p grit a x , the equivalent removal depth of a single grit. , ( ) p finial a x , the equivalent removal depth of final single grit. bmn, the offset of grain trajectory relative to the contact area center point. bmn, the offset of grain trajectory relative to the contact area center point. Bw, the width of the workpiece. Bc, the width of the contact wheel. B, the width of the contact area. ERub, the Young’s modulus of the contact wheel rubber outer layer. EAl, the Young’s modulus of the contact wheel aluminum inner core. E1, the Young’s modulus of the contact wheel. E1, the Young’s modulus of the contact wheel. E2, the Young’s modulus of the workpiece. E2, the Young’s modulus of the workpiece. E, the equivalent Young’s modulus. E, the equivalent Young’s modulus. eRa, the arithmetic average deviation error. eRa, the arithmetic average deviation error. eRt, the maximum altitude error . f0, the frequency of the belt machine tool driving wheel. Fgrit(x), the normal force of a single grain. Abstract Fn, the normal force between the workpiece and the contact wheel. hg, the height of a single grain. hs, the height of the gullies on grinded surface. hs, the height of the gullies on grinded surface. hd(x), the workpiece contour approximate equation in the contact area. hd(x), the workpiece contour approximate equation in the contact area. HV, the Vicker’s hardness of the workpiece. Hu, the ungrinded surface height. Hg, the grinded surface height. km, the coefficient of grit morphology. kp, the coefficient of material removal. kw, the coefficient of grit wear. kh, the coefficient of surface topography. kh, the coefficient of surface topography. kRa, the coefficient of arithmetic average deviation value caused by the stacking removal. kRt, the coefficient of maximum altitude value caused by the stacking removal. ps(x,y), the contact area pressure distribution in static contact condition. pd(x,y), the contact area pressure distribution in dynamic contact condition. pd(x,y), the contact area pressure distribution in dynamic contact condition. r, the radius of the contact wheel inner core. Ra,exp, the arithmetic average deviation value of the profile from experiments. Rt,exp, the maximum altitude value of the profile from experiments. Ra,ide, the ideal arithmetic average deviation value of the profile. Rt,ide, the ideal maximum altitude value of contour. Rt,ide, the ideal maximum altitude value of contour. Ra,pre, the arithmetic average deviation value of the profile from the prediction model. Rt,pre, the maximum altitude value of contour from the prediction model. Scont, the area of wheel-workpiece contact space. T, the type of grain size. vs, the linear velocity of the belt. vf, the robotic feed speed. vm, the equivalent cutting speed. v1, the Poisson ratio of the contact wheel. v2, the Poisson ratio of the workpiece.. δmax, the maximum deformation value of the contact wheel in the contact area. δs(x,y), the contact area deformation in static contact condition. δd(x,y), the contact area deformation in dynamic contact condition. ψ, the angle between robotic feeding speed orientation and cutting velocity orientation φ, the angle between grains arrangement orientation and robotic feed orientation. Δls, the width of the gullies on grinded surface Δlx, the horizontal spacing of grains Δly, the vertical spacing of grains. 1 Introduction The developments of advanced manufacturing industries, such as aerospace, new energy power generation, new energy vehicles, promote the demands of manufacturing components with free-form surfaces, just as engine blades, fan blades, wheel hubs and new energy vehicle bodies. In contemporary industrial productions, artificial works and special machine tools are the main craft for the preparation of parts with curved surfaces. These products fabricated by the manual grinding are sick in the rate of producing and qualities among different batches. While, manufacturing with special machine tools also takes disadvantages like the cost of equipment procurements. Robot assisted abrasive belt grinding is one species of free-form surface fabricating method with enormous potentials. Because of its structural characteristics, the stiffness of the serial robots is much lower than the one of the machine tools. While the elastic contact wheels of belt machine tools can provide passive flexibility which can reduce the deformation of robots in machining process. Nevertheless, the elastic deformation of the contact wheel complicates the grain trajectories in the removal process. The removal process and surface forming mechanism of robot assisted belt grinding is widely investigated [1-2]. For composite curved surface details with the requirement of dynamic characteristics, like blades roughness is an important property index of the qualities in service conditions. The threshold exceeded surface roughness causes air turbulent at the blade edge, resulting in the reduction of blade dynamic performance, and power losses of engine. Enormous surface roughness may cause cracks or even fracture due to the stress concentration effects, and ruined the performances in high temperature service conditions [3-5]. At present, the universal method for predicting the rigidly grinding surface roughness is based on the undeformed chip thickness model. The maximum undeformed chip thickness (MUCT) is a comprehensive coefficient which integrates both the cutting parameters and the grain status [6-8]. While the MUCT method is unable to predict exact value for elastic contact grinding procedures, such as belt grinding. Though the error can be reduced by introducing some elastic contact coefficients, the MUCT method is deficient in theoretical basis and the trend of MUCT predicted values is inconsistent with the experimental values [9]. Therefore, it is profitable to intensively investigate the removal mechanism of the elastic contact between abrasive grains and workpiece surfaces in elastic grinding process, which includes scratching, ploughing and cutting steps. Zhu et al. 1 Introduction evaluated the mechanism of robotic belt grinding by the perspective of grinding force [10]. The accomplishment of their work is the settlement of the micro cutting force model compensated with sliding, ploughing and cutting components. Based on the force model, the influence of force components on the machined surface roughness is discussed. Yang et al. instituted an energy efficiency model based on the friction coefficient model of a single spherical grain from the perspective of abrasive geometry [11], which showed that the ploughing energy took more proportion than the cutting energy and the scratching energy, while the grinding depth of a single particle was much smaller than the radius of grains. Based on the mechanism evaluation of robotic belt grinding, a noval grinding force model is accomplished by Xu et al. to predict the grinding removal depth and profit a quantificational machining process of robotic belt grinding [12,13]. Agustina and Segreto appraised the surface roughness obtained by robot-assisted polishing experiments with the analysis of the acoustic emission signal frequency domain features [14,15]. Zou et al. established a surface quality model considering the no-linear characteristic of rubber [16]. Pamdiyan et al. proposed an on-line multi-sensor integration system, which was composed with force sensors, acceleration sensors and acoustic emission sensors, to predict the surface roughness formed by robotic belt grinding through the neural network method [17]. In this paper, a noval surface topography model for surface roughness prediction in robotic belt grinding procedure considering the dynamic deformation of the elastic contact wheel is provided. With the infrastructures of the elastic plane-cylinder contact model and Preston equation, an original distribution of the contact pressure considering the dynamically removal volume during the elastic grinding procedure is attained. According to the plastic contact theory, the motional removal depth of a single pyramidal grain is gained by its relationship with the normal force of the grain. On the basis of the study in the equivalent removal depth of a single grit and the trajectories of grits in elastic grinding procedure an ideal surface topography model considering the effects of grain sizes is accomplished. Based on the topography model, the surface roughness obtained by robotic belt grinding can be predicted by introducing coefficients which means the effects of stacking removal phenomena. Through the robotic belt grinding experiments, the accuracy of the roughness predicting value is proved by comparing with the values in experiments. 2 The dynamic model and surface topography model involved in robotic belt grinding The workpiece settled on the actuator at the end of the robot is grinded by the belt grinding machine tool. During the grinding procedure, caused by the contact force between the contact wheel and the workpiece, deformations happen on both of the contact wheel and the workpiece while the one of the contact wheel is greater. Since getting into the contact area, normally, the grains adhered on the belt suffer three steps: the scratching step, the ploughing step and the cutting step. Figure1. The removing mechanism of the robotic belt grinding d i d l f Figure1. The removing mechanism of the robotic belt grinding 2.1 The dynamic model of contact pressure The normal force which generates the material removal is administered by the contact between the workpiece and the contact wheel. The contact wheel is mainly composed of an aluminum inner core and a rubber outer layer. In the research of the contact state, the deformation of the abrasive belt is ignored as for its thickness is much smaller than the radius of the contact wheel, while the diagrammatic sketch of the status of the contact interfaces is shown in Fig.1, which means both of the aluminum inner core and rubber outer layer are compressed under stress [9]. Therefore, the composite Young’s modulus of the contact wheel is crucial in establishing the contact model between the workpiece and the contact wheel. The combined Young’s modulus of contact wheel with considering the deformations of the inner and the outer is shown as Eq.1 and Eq.2. 1 Al Rub r E r r R r E R E    (1) ( ) = + = Al Rub n n c in out cont cont F r F r R r S E rS E      (2) 1 Al Rub r E r r R r E R E    (1) ( ) = + = Al Rub n n c in out cont cont F r F r R r S E rS E      (2) 1 Al Rub r E r r R r E R E    (1) ( ) = + = Al Rub n n c in out cont cont F r F r R r S E rS E      (2) In the robotic abrasive belt grinding procedure, both of the flexible contact wheel and the workpiece elastic take deformation. Though the elastic characteristic of rubber is non-linear, as for the elastic deformation is much smaller than the thickness of the outer layer, the linear elastic model can be used to calculate the rubber deformation. It is assumed that there is no relative motion between the wheel and the workpiece, which means that the contact is static. The contact wheel is abstracted as an elastic cylinder, while the workpiece is simplified as an elastic plane. According to the elastic cylinder-plane contact model [18], the normal contact force Fn is shown as Eq.3. 2.1 The dynamic model of contact pressure max, min( , ) 4 n w c F E B B B B     (3) * max, min( , ) 4 n w c F E B B B B     (3) The equivalent Young’s modulus E* is shown in Eq.4. The equivalent Young’s modulus E* is shown in Eq.4. 2 2 1 2 * 1 2 1 1 1 v v E E E     (4) 2 2 1 2 * 1 2 1 1 1 v v E E E     (4) The length of the static contact area between the workpiece and the contact wheel is expressed as Eq.5. max 2 sa R  (5) (5) The static contact area express as Eq.6. The static contact area express as Eq.6. max 2 cont S B R  (6) max 2 cont S B R  (6) max 2 cont S B R  (6) The static pressure ps(x,y)at an arbitrary point P(x,y) in the contact area is shown in Eq.7. 1 * 2 s( , ) 1 ( ) n s E F x p x y BR a          (7) Figure 2. The formation of contact area Th d f ti t P( ) i th t t i h i E 8 1 * 2 s( , ) 1 ( ) n s E F x p x y BR a          Figure 2. The formation of contact area The deformation at P(x,y) in the contact area is shown in Eq.8. The deformation at P(x,y) in the contact area is shown in Eq.8. 1 2 max ( , ) 1 ( ) s s x x y a           (8) At present, the static contact models, such as Hertz contact model or elastic cylinder-plane contact model, are generally used to predict the material removal volume in robotic belt grinding. The static contact models suppose that the contact parameters, such as contact area and contact stress distribution, are independent with material removal volume in grinding process. However, in the actual manufacturing, due to the grinding removal effect of the adhesive grits, the actual contact surface between the wheel and the workpiece is not an ideal horizontal plane. 2.1 The dynamic model of contact pressure , p n m p total w k F v a v  (11) , p n m p total w k F v a v  (11) In the grinding procedure，the length of the dynamic contact area ad is expressed as Eq.12 In the grinding procedure，the length of the dynamic contact area ad is expressed as Eq.12   2 2 max max , , + ( ) + d p total p total a R R a a     max max , + ( ) p total R R a     (12) The workpiece contour in contact area is fitted with a straight line, as shown in Fig.3, the approximate equation hd(x) is shown as Eq.13, , max max , max , , ( ) ( ) ( ( )- p total d p total p total p total a h x R R a x R a a          （） (13) where，the equivalent cutting speed vm is shown as Eq.14 0 0 , 2 m s w s v v v v R f     (14) 0 0 , 2 m s w s v v v v R f     (14) 0 0 , 2 m s w s v v v v R f     (14) where，vs is the linear velocity of the belt，vw is the feed speed of the robot, ‘+/-’ means that the linear velocity of contact wheel and the workpiece feed speed takes the same/opposite direction. (b) Dynamic contact model (a)Static contact model (b) Dynamic contact model Figure 3. Model of static and dynamic contact condition (a)Static contact model (a)Static contact model (b) Dynamic contact model Figure 3. Model of static and dynamic contact condition Figure 3. Model of static and dynamic contact condition Figure 3. Model of static and dynamic contact condition Due to the Young’s modulus of the workpiece is much larger than that of the contact wheel, it can be regarded as that only the contact wheel takes the elastic deformation, while the deformation of the workpiece is zero, which is shown as Eq.15. 2.1 The dynamic model of contact pressure As Fig.2 shows, the leading edge of the actual contact area is the ungrinded surface, while the training edge is the grinded surface. The height of workpiece in contact area is analyzed. In the coordinate system of the contact area OXYZ, which the orientation of x axis is the robotic feed direction, y axis is perpendicular to x axis and plane XOY is parallel to ungrinded workpiece surface, z axis is perpendicular to x and y axis. 1 1 2 2 ( ) 0, ( ) 0 d d x x x x         (9) 1 1 2 2 ( ) 0, ( ) 0 d d x x x x         (9) As x=x1, the height of the workpiece is the ungrinded surface height Hu，while as x=x2, is the grinded surface height Hg, which is expressed as Eq.10. , u g p total H H a   (10) where，ap,total is the totally removal depth，which can be obtained by Preston Equation, as Eq.11 [19]. 2.1 The dynamic model of contact pressure The abrasive particles trajectory equation on the contact area plane XOY is shown as Eq.19 [20 p j y q p q [ ] tan + mn y x b    (19) Figure 4. The grit morphology characteristics where，ψ is the angle between the robotic feeding orientation and the cutting velocity orientation，bmn is the offset of tan + mn y x b    (19) tan + mn y x b    (19) Figure 4. The grit morphology characteristics Figure 4. The grit morphology characteristics where，ψ is the angle between the robotic feeding orientation and the cutting velocity orientation，bmn is the offset of grain trajectory relative to the contact area center point. where，ψ is the angle between the robotic feeding orientation and the cutting velocity orientation，bmn is the offset of grain trajectory relative to the contact area center point. The normal force of a single grit Fgrit(x) is expressed as Eq.20. The normal force of a single grit Fgrit(x) is expressed as Eq.20. 1 1 2 2 1 1 2 2 ( ) ( ) x y x y x l x l grit d x l y l F x p x dydx       (20) Wang used the material hardness to calculate the removal depth of a single grain [21].Zhang discovered that the grain wear condition influenced material removal depth[22]. Asikuzun provided the relationship between Vickers hardness, normal force and indentation depth [23]. As the grain morphology is similar with the Vickers hardness testing penetrator, the removal depth of a single grain ap,grit(x) can be calculated as Eq.21. , 2 ( ) ( ) Hv tan 2 grit p g m rit w F x a x k k    (21) , 2 ( ) ( ) Hv tan 2 grit p g m rit w F x a x k k    (21) where, kw is the coefficient of grain wear. km is the coefficient of abrasive grit morphology as Eq.22 shown. ==0.9032 0.9594 m k  (22) When the pressure is less than 0.4HB, only elastic deformation occurs [24]. As Eq.21 shows when the grains are abstracts as ideal pyramids, the pressure between the grain and the workpiece is HV 0.4HB grit p   , so in this 2.1 The dynamic model of contact pressure =0 = ( ) w c x       (15) =0 = ( ) w c x       (15) The removal volume coefficient kr,(x), expressed as Eq.16, indicates the reduction of the contact wheel compression caused by material removal in the stable cutting procedure. ( ) ( )+ ( ) ( ) = ( ) ( ) d s d r s s x x h x k x x x      (16) ( ) ( )+ ( ) ( ) = ( ) ( ) d s d r s s x x h x k x x x      (16) In the dynamic contact model，the dynamic pressure pd(x,y) at an arbitrary point P(x,y) in the contact area is shown in Eq.17. In the dynamic contact model，the dynamic pressure pd(x,y) at an arbitrary point P(x,y) in the contact area is shown in Eq.17. 1 * 2 ( )+ ( ) ( , ) 1 ( ) ( ) s d n d s s x h x E F x p x y x BR a            (17) 1 * 2 ( )+ ( ) ( , ) 1 ( ) ( ) s d n d s s x h x E F x p x y x BR a            (17) 2.2 The single grain grinding depth and material removal volume 2.2 The single grain grinding depth and material removal volume particle morphology, such as the height of abrasive grits hgrit, the horizontal spacing between two adjacent abrasive grits is Δlx, vertical spacing Δly, are attained. The angle between the relative surfaces θ is shown in Eq.18. particle morphology, such as the height of abrasive grits hgrit, the horizontal spacing between two adjacent abrasive grits is Δlx, vertical spacing Δly, are attained. The angle between the relative surfaces θ is shown in Eq.18. tan = 2 2 x g l h   (18) tan = 2 2 x g l h   (18) The abrasive particles trajectory equation on the contact area plane XOY is shown as Eq.19 [20]. 2.3 The surface topography model The removal width of a single grit is much smaller than the horizontal distance between two adjacent abrasive grits Δlx. Actually，the morphology of workpiece surface is formed by the grinding tracks of different grains at different times. In order to express the vertical positions of different grains reaching the cross section, the equivalent cutting depth of single abrasive particles is introduced as Eq.23. equivalent cutting depth of single abrasive particles is introduced as Eq.23. equivalent cutting depth of single abrasive particles is introduced as Eq.23. 2 , , , ( ) ( ) ( ) 2 cot( ) 2 p grit p grit p grit x x S x a x a x l l      (23) 2 , , , ( ) ( ) ( ) 2 cot( ) 2 p grit p grit p grit x x S x a x a x l l      (23) Since the removal depth of single grit is much smaller than the elastic contact deformation, the equivalent removal depth of a single abrasive grain , ( ) p i a x , shown as Eq.24, can be regarded as a constant volume in the calculation of the robotic belt grinding surface roughness. 2 , , ( ) ( ) 2 cot( ) 2 p Nq p finial x a x a x l    (24) 2 , , ( ) ( ) 2 cot( ) 2 p Nq p finial x a x a x l    (24) As for the angle φ between the grain arrangement direction and the robot feed direction，the horizontal distance between the locations which are trajectories of two adjacent grains in the same row in cross section is Δly×sin(φ)， while the vertical one is , - ( ) p finial a x . The points in the area of {(xsection,ysection)\| 0<xsection≤Δlx , -ap,N(x)<ysection≤0} can be expressed with (xm,ym) as Eq.5 al one is , - ( ) p finial a x . 2.3 The surface topography model 0 max min 1 l a t R ydx l R y y         (27) 0 max min 1 l a t R ydx l R y y         (27) The ideal surface roughness along the robot feed direction of belt grinding can be obtained as Eq.28. The ideal surface roughness along the robot feed direction of belt grinding can be obtained as Eq.28. , , , , ( ) = 4 4 = ( ) h p Nq s a ide t ide s h p Nq k a x h R R h k a x       (28) , , , , ( ) = 4 4 = ( ) h p Nq s a ide t ide s h p Nq k a x h R R h k a x       (28) In the surface profile model establishing process, the relationship between different trajectories of grits is assumed to be the parallel removal mode, which means that the adjacent trajectory is approximately horizontally arranged. The adjacent removal mode is shown in Fig.6. Nevertheless, in the actual surface forming process, besides the adjacent removal mode, the stacking removal mode is also exist. The stacking mode means that the adjacent grinding tracks overlap in the vertical direction, therefore the grinding depth stacking one by one, which makes the gully depth on the surface profile exceeding the ideal one hs. The stacking removal mode is shown in Fig.6. Figure 6. The adjacent removal mode and the stacking removal mode Figure 6. The adjacent removal mode and the stacking removal mode Figure 6. The adjacent removal mode and the stacking removal mode Through the analysis of the robotic belt grinding surface topography, it can be found that the adjacent removal mode is the main stream of material removing. Contrarily, the stacking removal mode occurs randomly, while the maximum removal depth formed by the stacking removal mode has a positive correlation with the single grit removal depth. 2.3 The surface topography model The points in the area of {(xsection,ysection)\| 0<xsection≤Δlx , -ap,N(x)<ysection≤0} , , rem( sin( ) , ) , ( ) 2 cot( ) 2 1,2..., ( ) m y x p finial m x p Nq x m l l y m a x l m a x                       (25) , , rem( sin( ) , ) , ( ) 2 cot( ) 2 1,2..., ( ) m y x p finial m x p Nq x m l l y m a x l m a x                       (25) where，Rem() is the remainder function. Figure 5. The ideal surface topography formed by robotic belt grinding The surface formed by the grain tracks are shown as Fig.5. The grinded surface morphology, which is the blue line in Fig.6, is the envelope of abrasive trajectories. Δls is the width of the gullies on grinded surface，while hs is the height, expressed as Eq.26. where，Rem() is the remainder function. Figure 5. The ideal surface topography formed by robotic belt grinding The surface formed by the grain tracks are shown as Fig.5. The grinded surface morphology, which is the blue line in Fig.6, is the envelope of abrasive trajectories. Δls is the width of the gullies on grinded surface，while hs is the height, expressed as Eq.26. The surface formed by the grain tracks are shown as Fig.5. The grinded surface morphology, which is the blue line in Fig.6, is the envelope of abrasive trajectories. Δls is the width of the gullies on grinded surface，while hs is the height, expressed as Eq.26. , (0) s h p Nq h k a  (26) where，kh is the surface topography coefficient. deviation value of the profile Ra and the maximum altitude value of contour Rt, calculated as Eq.27. deviation value of the profile Ra and the maximum altitude value of contour Rt, calculated as Eq.27. 2.3 The surface topography model Above all, the robotic belt grinding surface roughness of can be predicted as Eq.29, , , , , , e , ( ) = 4 = ( ) h Ra p Nq a pre Ra a ide t pre Rt t id h Rt p Nq k k a x R k R R k R k k a x       (29) , , , , , e , ( ) = 4 = ( ) h Ra p Nq a pre Ra a ide t pre Rt t id h Rt p Nq k k a x R k R R k R k k a x       (29) where, kRa is the coefficient of arithmetic average deviation value caused by the stacking removal，kRt is the one of maximum altitude value. where, kRa is the coefficient of arithmetic average deviation value caused by the stacking removal，kRt is the one of maximum altitude value. 3.1 Experimental conditions The morphologies of abrasive grits The grinding system characteristics in the belt grinding surface roughness model are shown in Tab.2. 3.2 Experimental design ng system characteristics in the belt grinding surface roughness model are shown in Tab.2. The experimental parameters were selected as Tab.3 After the tests, the surface morphologies of the workpieces were scanned by the laser confocal microscope, which could calculated the surface roughness value obtained by robotic belt grinding. Table 3. Grinding parameters of experiments Table 3. Grinding parameters of experiments Grinding parameters Value ranges Type of grain size T (#) 30,45,65,80,100 Driving wheel frequency f0(Hz) 5,10,15,20,25 Robotic feed speed vw(mm/s) 1,3,5,7,9 Maximum contact depth δmax(μm) 20,40,60,80,100 4 ifi i i i Robotic feed speed vw(mm/s) Maximum contact depth δmax(μm) 3.1 Experimental conditions All the robotic belt grinding experiments were conducted on the robotic belt grinding system mainly composed of a 6-DOF robot with the type of Motorman DX100, as shown in Fig.7. The Inconel 718 superalloy samples which were cut to 500mm(length) × 15mm(width) × 10 mm(thickness) pieces were fixed with the specific fixtures. The belt 237AA (3M Company), covered with Al2O3 grains, were used in the experiments. The micro-morphologies of the grains were shown by KEYENCE VHX-1000C digital microscope as Fig.8, which verified the grain morphology model. Table 1. Grains morphology characteristics Type of grain size T(#) Grain height hg(um) Grains vertical spacing Δlx(um) Grains horizontal spacing Δly (um) 30 247 392 411 45 291 467 450 65 272 508 472 80 296 586 526 100 389 713 680 Table 2. Grinding system characteristics Items Conditions Contact Wheel r=50mm, R=100mm, R0=100mm, EAl=70Gpa, ERub=7.8Mpa, v1=0.47, Bc=50mm, ψ=0°, φ=14°, bmn=0 Workpiece E2=199.9GPa, v2=0.3, Hv=3.376Gpa, Bw=50mm, kw=1.8, kRa=1, kRt=2.1 Figure 7.The robotic belt grinding system The abrasive grit morphology characteristics of each type of abrasive belts are shown in Tab.1. Table 1. Grains morphology characteristics Type of grain size T(#) Grain height hg(um) Grains vertical spacing Δlx(um) Grains horizontal spacing Δly (um) 30 247 392 411 45 291 467 450 65 272 508 472 80 296 586 526 100 389 713 680 Table 2. Grinding system characteristics Items Conditions Contact Wheel r=50mm, R=100mm, R0=100mm, EAl=70Gpa, ERub=7.8Mpa, v1=0.47, Bc=50mm, ψ=0°, φ=14°, bmn=0 Workpiece E2=199.9GPa, v2=0.3, Hv=3.376Gpa, Bw=50mm, kw=1.8, kRa=1, kRt=2.1 Figure 7.The robotic belt grinding system The abrasive grit morphology characteristics of each type of abrasive belts are shown in Tab.1. ( ) The abrasi e grain micro morpholog E2=199.9GPa, v2=0.3, Hv=3.376Gpa, Bw=50mm, kw=1.8, kRa=1, kRt=2.1 Figure 7.The robotic belt grinding system The abrasive grit morphology characteristics of each type of abrasive belts are s Figure 7.The robotic belt grinding system The abrasive grit morphology characteristics of each type of abrasive belts are shown in Tab.1. The abrasive grit morphology characteristics of each type of abrasive belts (a) The abrasive grain micro-morphology (a) The abrasive grain micro-morphology (a) The abrasive grain micro-morphology (b) The different wear condition grain morphology model Figure 8. The morphologies of abrasive grits The grinding system characteristics in the belt grinding surface roughness model are shown in Tab.2. (b) The different wear condition grain morphology model Figure 8. 4. Verification and discussion For the sake of verifying the belt grinding surface roughness mode, Eq.29 was used to calculate the surface roughness predicted value which are compared with the experimental values to characterize the accuracy of the surface roughness model. The arithmetic average deviation error eRa is 11.6% while the maximum altitude error eRt is 7.03% respectively, which are calculated as Eq.22 and Eq.2 Eq.23.Through error analysis, the efficiency of the surface roughness model is verified. Compared with the normal static contact surface roughness model, the improved dynamic contact surface roughness model provides higher precisions. exp ,exp , 1 ,exp exp 100% i n a a pre i a Ra R R R e n       (30) exp ,exp , 1 ,exp exp 100% i n t t pre i t Rt R R R e n       (31) exp ,exp , 1 ,exp exp 100% i n a a pre i a Ra R R R e n       (30) exp ,exp , 1 ,exp exp 100% i n t t pre i t Rt R R R e n       (31) Fig.9 shows the relationships between the surface roughness and the grinding parameters. As shown in Fig.9(a), when the belt linear velocity is under a threshold, the surface roughness model provides an accurate prediction. When the velocity is over the threshold, the surface roughness is much larger than the predicted value. It is caused by the elastic recoil phenomenon of the rubber layer [25]. Both the simulations and the experiments suggest that the surface roughness get deteriorated with the increasing of contact depth, as Fig.9(b) shown, while the feeding speed only makes a slight effect on surface roughness as Fig.9(c) shown. It is observed that the grain size and contact depth have great influence on the surface quality as Fig.9(d) shows. The abrasive particle size has the most significant effect on the surface roughness, and the surface roughness reaches over Ra0.6 when the type of grain is #100, while the surface roughness is reduced under Ra0.3 with #30. This conclusion is supported by Gorp AV [26], that the most important factor affecting the workpiece roughness is the grain size. roughness is the grain size. roughness is the grain size. 4. Verification and discussion (a) (b) (c) (d) (a)The relationship between the surface roughness and the belt cutting velocity; (b) the relationship between the surface roughness and the maximum contact depth; (c) the relationship between the surface roughness and the robotic feed speed; (d) the relationship between the surface roughness and the size of grains Fi 9 Th l ti hi b t th f h d th i di t (b) (a) (b) (c) (d) (d) (c) (a)The relationship between the surface roughness and the belt cutting velocity; (b) the relationship between the surface roughness and the maximum contact depth; (c) the relationship between the surface roughness and the robotic feed speed; (d) the relationship between the surface roughness and the size of grains Figure 9. The relationships between the surface roughness and the grinding parameter Figure 9. The relationships between the surface roughness and the grinding parameter 5. Conclusion In this study, a novel surface topography model of robotic belt grinding procedure considering the dynamic deformation of the elastic contact wheel is proposed. Based on the elastic plane-cylinder contact model and Preston equation, an original distribution of the contact pressure considering the dynamically removal volume during the elastic grinding procedure is attained. Besides, on the basis of the study in the equivalent removal depth of a single grit and the trajectories of grits in elastic grinding procedure, an ideal surface topography model considering the effects of grain sizes is accomplished. The results indicated that the surface topography formed by elastic grinding related to the removal depth of a single grain, unlike the one formed by rigid grinding which is effected by the totally removal depth. Moreover, the surface roughness forecast model is provided by considering the removal depth of a single grain and the trajectory of active grains in robotic belt grinding process, and the verified experimental results of robotic belt grinding disclosed that the prediction error can be confined to 11.6%, which can predict the surface topography information of robotic belt grinding procedure more accurately compared with the traditional predicted models. References [1]Zhu D, Feng X, Xu X (2020) Robotic grinding of complex components: A step towards efficient and intelligent machining – challenges, solutions, and applications[J]. Robot. Cim-Int. Manuf. 65:101908. https://doi.org/10.1016/ j.rcim. 2019.101908 [1]Zhu D, Feng X, Xu X (2020) Robotic grinding of complex components: A step towards efficient and intelligent machining – challenges, solutions, and applications[J]. Robot. Cim-Int. 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Int J Adv Manuf Tech 104:3209–3217. https://doi.org/10.1007/s00170-019- 04332-7 [10]Zhu DH, Xu XH, Yang ZY (2018) Analysis and assessment of robotic belt grinding mechanisms by force modeling and force control experiments. Tribol Int 120:93–98 https://doi.org/ 10.1016/j.tribo int.2017.12.043 [11]Yang ZY, Xu XH, Zhu DH (2019) On energetic evaluation of robotic belt grinding mechanisms based on single spherical abrasive grain model. Int J Adv Manuf Technol 104:4539–4548 https://doi.org/10.1007/s00170-019- 04222-y [11]Yang ZY, Xu XH, Zhu DH (2019) On energetic evaluation of robotic belt grinding mechanisms based on single spherical abrasive grain model. Declaration Acknowledgments Funding The authors would like to acknowledgement financial support from the National Natural Science Foundation of China Key (Joint Fund) Project (no. U1908230) Conflicts of interest/Competing interests The authors declare no competing interests. Availability of data and material The datasets used or analyzed during the current study are available from the corresponding author on reasonable request. Ethics approval The authors state that the present work is in compliance with the ethical standards. Consent to participate Not applicable. Ethics approval The authors state that the present work is in compliance with the ethical standards. Consent to participate Not applicable. Consent for publication All authors agree to publish. Authors' contributions Mingjun Liu: Investigation, Conceptualization, Methodology, Experiment, Writing - original draft. Yadong Gong: Funding and acquisition, reviewed & edited the manuscript, Supervision. Jingyu Sun: Investigation, Experiment. Yuxin Zhao: Investigation, Experiment. Yao Sun: Supervision. References Authors' contributions Mingjun Liu: Investigation, Conceptualization, Methodology, Experiment, Writing - original draft. 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J Mater Sci-Mater El 23(5):1001-1010 https://doi.org/10.1007/s10854 -011-0537-0 -011-0537-0 [24]Zhao YW, D.M. Maietta, Chang L (2000) An asperity microcontact model incorporating the transition from elastic deformation to fully plastic flow. J Tribol-T Asme 122:86-93 https://doi.org/ 10.1115/1.555332 [25]Yang ZY, Chu Y, Xu XH (2021) Prediction and analysis of material removal characteristics for robotic belt grinding based on single spherical abrasive grain model. Int J Mech Sci 190 https://doi.org/10.1016/j.ijmecsci.2020.106005 [26]Gorp A V, Bigerelle M, Mansori M E (2015) Effects of working parameters on the surface roughness in belt grinding process: the size-scale estimation influence. Int J Mater Prod Tec 38(1). https://doi.org/10.1504/ijmpt. 2010.031892
https://openalex.org/W2081948122	https://periodicos.ufsc.br/index.php/mundosdotrabalho/article/download/1984-9222.2013v5n10p261/26763	Portuguese	null	LEAL, Murilo. A Reinvenção da classe trabalhadora (1953-1964). Campinas: Editora da Unicamp, 2011, 520 p.	Revista Mundos do Trabalho	2,013	cc-by	2,831	∗ Luciana Wollmann é doutoranda pelo Programa de Pós-Graduação em História Política e Bens Culturais do CPDOC/ FGV. 1 BORGES apud FONTES, Paulo. Um nordeste em São Paulo. Rio de Janeiro: FGV, 2008, p. 47. 2 Tomo de empréstimo aqui a metáfora utilizada por Peter Linebaugh e Marcus Redikersobre a formação do proletariado atlântico em: A hidra de muitas cabeças: marinheiros, escravos e plebeus e a história oculta do Atlântico revolucionário. São Paulo: Cia. das Letras, 2008. http://dx.doi.org/10.5007/1984-9222.2013v5n10p261 http://dx.doi.org/10.5007/1984-9222.2013v5n10p261 Luciana Wollmann∗ LEAL, Murilo. A Reinvenção da classe trabalhadora (1953-1964). Campinas: Editora da Unicamp, 2011, 520 p. ∗ Luciana Wollmann é doutoranda pelo Programa de Pós-Graduação em História Política e Bens Culturais do CPDOC/ FGV. A classe trabalhadora para além Luciana Wollmann∗ Palavras-chave: Trabalhadores ― Populismo ― Sindicalismo Palavras-chave: Trabalhadores ― Populismo ― Sindicalismo Keywords: Workers ― Populism ― Trade-unionism 2 Tomo de empréstimo aqui a metáfora utilizada por Peter Linebaugh e Marcus Redikersobre a formação do proletariado atlântico em: A hidra de muitas cabeças: marinheiros, escravos e plebeus e a história oculta do Atlântico revolucionário. São Paulo: Cia. das Letras, 2008. 1 BORGES apud FONTES, Paulo. Um nordeste em São Paulo. Rio de Janeiro: FGV, 2008, p. 47. Keywords: Workers ― Populism ― Trade-unionism Essa contraposição mostrou-se essencial para o autor demonstrar os diferentes impactos experimentados por esses setores, a partir do processo de “reestruturação produtiva” naqueles que foram considerados os “anos áureos” do nacional-desenvolvimentismo. Para além dos impactos “na esfe ra da produção de mercadorias”, as interferências desses processos nos espaços de reprodução social foram também cuidadosamente esmiuçadas pelo autor. É verdade que a análise de Leal “[...] vai muito além do exame de duas ca tegorias” (p. 20). Temas abrangentes como populismo, trabalhismo, sindicalismo pré-1964, padrões de urbanização e experiências migratórias ocupam lugar desta cado no trabalho do autor e são tomados sobre perspectivas tão originais quanto desafiadoras. Descortinando debates clássicos da historiografia brasileira, ele nos coloca questões que, embora não explicitamente formuladas, se encontram pre sentes, de uma maneira ou de outra, em toda a extensão da sua obra: qual foi o peso do papel social, político e econômico desempenhado pela classe trabalhado ra e pelas demais classes populares no período da chamada “República Populis ta”? É possível compreendê-los apenas como porção mais frágil de um “pacto” ― populista ou trabalhista? Ou eles estariam envolvidos em um complexo, mutável e desigual sistema de alianças? Se sim, que alianças seriam essas? Por quem e de que maneira elas foram compostas? Foi possível para a classe trabalhadora formu lar um projeto contra-hegemônico nesse período? Sim? Não? Quais foram os seus mediadores? Apresentando uma ampla e variada gama de fontes analisadas a par tir de um escopo teórico e empírico consistente, o autor nos fornece elementos importantes para esses debates. 262 Elegendo como marco inicial o ano de 1953 ― ano da greve dos 300 mil e da primeira eleição municipal realizada após a Revolução de 1930 ― e como ano final o golpe civil-militar de 1964, o estudo de Leal procura pensar como a conquista do espaço público experimentado pelos trabalhadores a partir das greves apresen tou-os como sujeito coletivo, sendo esse um dos aspectos chave para a sua análise sobre a “reinvenção” de classe. Todavia, ao destacar o processo eleitoral como justificativa para o seu recorte cronológico, o autor não sugere que este “refazer -se” se processou de forma “autônoma” ou “espontânea”, e nem tampouco de maneira “orquestrada” a partir da “cooptação” de alguns líderes. Keywords: Workers ― Populism ― Trade-unionism Josés, Marias, Joões, milhares de Ulisses. Destino: São Paulo. A cidade, que fora ponto de partida dos “heróis nacionais” que desbravaram os recônditos mais longínquos do Brasil em busca de riquezas no século XVII, passou a receber nos anos de 1950, uma infinidade de “heróis anônimos” (p. 42) que também cultiva vam, em si, a ideia do progresso. Oriundos do interior paulista, de Minas Gerais e, sobretudo, de diferentes estados do Nordeste brasileiro, eles eram trabalhadores e trabalhadoras que, motivados pelo ideário de riqueza e trabalho ― cada vez mais presentes nas narrativas sobre a capital paulista daquele período (p. 19) ― procuravam melhores oportunidades naquela que já era considerada a “Meca” dos migrantes.1 As “ilusões da cidade” (p. 48), no entanto, não tardaram em apa recer. Não tão prestigiados quanto os heróis de outrora, os protagonistas dessa epopeia foram estimulados a ocupar novos territórios, localizados nas zonas pe riféricas da cidade. Num “esforço hercúleo” de enfrentar uma dupla jornada de trabalho ― nas fábricas e na confecção de suas moradas ―, esses trabalhadores foram responsáveis por imprimir novos traçados à cidade, ao mesmo tempo em que nela teciam seus laços. Tomando parte dessa “Pauliceia”, não demoraria mui to para que nossos heróis percebessem que em terra em que se poderia ser um tanto Hércules ou Ulisses poder-se-ia também ser “Hidra de muitas cabeças”.2 Em “A Reinvenção da classe trabalhadora (1953-1964)”, Murilo Leal nos convida a acompanhar a saga desses milhares de trabalhadores que, já vivendo na cidade ou passando a tomar parte dela com as “novas correntes migratórias” ocorridas entre os anos 1940 e 1960, se “aventuravam” cotidianamente naquela grande metrópole. Em um esforço bem sucedido de tentar “integrar dois níveis de análise” (p. 34) ― o individual e o coletivo ―, o autor elegeu duas categorias de trabalhadores para a sua investigação: metalúrgicos e têxteis. Tal escolha se justifica, não só por serem as “duas maiores categorias operárias da época”, como Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 EDILENE TOLEDO EDILENE TOLEDO esclarecem seus prefaciadores3, mas também por configurarem dois setores que se diferenciam, tanto por seus modelos de produção e capacidade de moderniza ção industrial quanto pela composição social, pela organização e pelas formas de luta de seus trabalhadores. 3 O livro é apresentado por Paulo Fontes e prefaciado por Marcelo Badaró. Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... expoentes da história social britânica, como E. P. Thompson e Eric Hobsbawm, e para a historiografia do trabalho brasileira recente. O diálogo com brasilianistas, com outros marxistas ― Raymond Williams, Antonio Gramsci e Henri Lefebvre ― e com autores que se dedicaram a pesquisar temas como populismo e trabalhismo tam bém merece ser destacado, pois eles estiveram presentes na análise do autor, seja como arcabouço conceitual ou como repertório para as suas formulações críticas. A ideia de “reinvenção”, que busca dialogar com a obra clássica de Angela de Castro Gomes4 e que também encontra lugar em alguns pressupostos da História Cultural, refere-se à classe, não como uma categoria fixa e imutável, mas como algo flexível que incorpora ou ressignifica elementos, crenças e valores em deter minadas conjunturas. Naquele período, segundo o autor, “[...] as próprias caracte rísticas da classe operária mudaram constantemente” e “[...] o repertório de cren ças e valores com os quais a classe se via no começo da década de 1950 era muito diferente daquele que aparece nos discursos no começo dos anos 1960” (p. 36). Na primeira parte do livro, o autor busca apresentar algumas características “estruturais” da industrialização e da urbanização periférica de São Paulo, obser vando não apenas como essas “condições objetivas” interferiram na experiência operária, mas também como os trabalhadores puderam lhe exercer influência. Duas categorias marxistas ― Produção material e Reprodução social ― fornecem inspiração aos títulos, ainda que a maior influência talvez advenha de uma releitu ra lefebvriana desses conceitos.5 Em “Espaços de Produção” (cap. 1), Leal procura refletir sobre o papel da classe trabalhadora nas transformações que ocorreram na industrialização brasileira a partir dos anos de 1950. O processo de migração, as características das indústrias metalúrgica e têxtil, o aprendizado profissional ― adquirido em escolas ou no “chão de fábrica” ―, a heterogeneidade do proleta riado brasileiro ― composto por homens, mulheres, jovens de diferentes origens ―, as diferentes formas de remuneração, as reivindicações por local de trabalho e as condições insalubres da produção fabril compuseram fatores agregadores e de sagregadores da classe trabalhadora. Debruçar sobre eles nos fornece elementos para repensar como a classe operária exerceu influência no projeto industrializante articulado “pela burguesia industrial e pela burocracia de Estado” (p. 45), ainda que estivesse “desarticulada em seu potencial político contra hegemônico” (Idem). 5 Pensando o espaço urbano como um elemento central do modo de produção capitalista, Henri Lefebvre analisa como a “produção do espaço” produz relações sociais, ao mesmo tempo em que são produzidas por elas. Essa ideia, central nas análises do autor, se encontra presente em suas diferentes obras. Murilo Leal faz referência a dois títulos, sendo eles: LEFEBVRE, Henri. La presencia y la ausência ― Contribuicíon a lateoría de las representaciones. Ciudad de Mexico: Fondo de Cultura Económica, 1983; LEFEBVRE, Henri. A vida cotidiana do mundo moderno. São Paulo: Ática, 1991. 4 GOMES, Angela de Castro Gomes. A invenção do trabalhismo. Rio de Janeiro: Relume Dumará, 1984. Keywords: Workers ― Populism ― Trade-unionism Para o autor, entenderemos melhor a dinâmica desse processo “[...] se substituirmos a ideia de anomia, heteronomia ou mesmo de ‘coletivo socialmente heterogêneo’ pela ideia de aliança de classes” (p. 115). O livro está dividido em três partes: Parte I: “Determinações” (capítulos 1 e 2); “Práticas: lutas reivindicações, organizações” (capítulos 3 a 8); Parte III: “Re presentações: cultura e política (capítulos 9 e 10)”. Ainda que o autor não abra mão de uma organização sequencial, que forneça ao leitor uma dimensão crono lógica dos acontecimentos ― indispensável no momento de análise das greves ―, observamos que são os aspectos concernentes à própria experiência dos trabalha dores os que mais interferem na disposição dos capítulos. Afinal de contas, Murilo Leal deixa claro logo na introdução a sua opção por uma “história vista de baixo”, assim como enumera os referenciais teóricos/ de análise que o auxiliaram a situar o seu trabalho a partir dessa perspectiva, com destaque para as contribuições de 3 O livro é apresentado por Paulo Fontes e prefaciado por Marcelo Badaró. Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... Os motins urbanos, os pi quetes, os abaixo-assinados e manifestos, as alianças estabelecidas e desfeitas, as intersindicais, as associações, as greves dos 300 mil em 1953 a dos 400 mil ― por aumento salarial e contra a carestia ― em 1957, a dos 700 mil ― considerada a mais ampla e organizada, mas menos popular ― em 1963, são analisados em detalhes e nos possibilitam visualizar como as demandas urbanas, de consumo e às referentes ao mundo do trabalho tiveram ressonância na voz de um “sujeito coletivo” que forjou para si, e com auxílio de outros atores, uma “esfera pública plebeia” (p. 168). Na esteira das lutas, a “justiça legal” e “moral” é acionada. A pu blicização da questão operária e popular por meio da imprensa sindical, comunista e popular, e de denúncias feitas à Câmara e à Assembleia Legislativa possibilitaram a divulgação e a legitimação da “condição operária”. Nesse ínterim, ao contrário de diluir-se no todo heterogêneo das “classes populares”, como afirmavam alguns dos expoentes da bibliografia crítica ao populismo, o autor identifica uma “hege monia proletária no campo popular” (p. 461). 264 264 Na terceira e última parte do livro, não por acaso, cultura operária e popu lar, representação política e consciência de classe são apresentadas em um só bloco. Afinal, “cultura” e “política” não são vistas aqui como “entes” separados do processo social, mas como elementos constitutivos que estão “[...] presos na mesma rede de relações”.6 No capítulo 9, o tempo de festas e de lutas marcado no calendário operário “[...] oferece-nos pistas sobre os valores compartilhados e significados atribuídos aos eventos” (p. 397). A labuta diária empreendida pelos trabalhadores contra os pequenos furtos do capital sobre o seu tempo (discutida na parte 2 do livro) era engendrada também no plano da cultura e dos costumes. O despotismo patronal, comumente relembrado por suas “raízes patriarcais e escravocratas” (p. 148-157), poderia, da mesma forma, ser enfrentado pelos tra balhadores a partir do controle do seu “tempo livre”. Logicamente que a “cultura operária” não é tomada pelo autor de forma estandardizada, pois, ainda que pos sua especificidades, se encontra entremeada por valores da cultura popular, pe las comemorações oficiais do Estado e até por elementos da sociedade patriarcal. UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... 263 263 No capítulo 2, “Espaços de Reprodução”, o autor analisa as bases e o desen volvimento daquilo que ele denominou “padrão periférico de habitação operária e popular”. A especulação imobiliária ocorrida nas áreas centrais da cidade transfe riu grandes contingentes de trabalhadores para as regiões mais afastadas. O “feti chismo da mercadoria” tomava, então, forma de habitação popular. Extraindo ho ras extras da vida dos trabalhadores ― tanto na esfera de produção fabril quanto na esfera de produção doméstica (p. 95) ―, o “padrão periférico” de urbanização das “áreas vazias” da cidade se deu a partir da superexploração do trabalho e da espoliação urbana. Porém, nem só de trabalho abstrato se fez a ocupação das pe riferias. O trabalho concreto dos mutirões organizados para construção das “casas domingueiras”, as redes de solidariedade tecidas, a realização do “sonho da casa própria” ― que “verdadeiro ou não” diminuía o sentimento de “insegurança es trutural” dos trabalhadores ― e, finalmente, a luta coletiva por direitos demons Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 EDILENE TOLEDO tram que, para além do “pacto” consubstanciado pelo consentimento do poder público diante das ocupações “irregulares” das periferias, a experiência de classe também se fez a partir de lutas sociais pelo direito à cidade. Enfraquecidos pela dis persão espacial de um lado, mas fortalecidos pelas “redes densas” e pelas “redes amplas” propiciadas pelos “mediadores espaciais” ― SABs, CDPs, imprensa diária e políticos que iam buscar apoio local ― por outro, o autor demonstra como o es paço também se apresenta como um elemento constitutivo da formação de classe. Na segunda parte ― a mais volumosa do livro ― o autor analisa separada mente as lutas fabris e de bairro para, posteriormente, demonstrar como as lu tas contra a carestia possibilitaram a formação de uma aliança interclassista, bem como a elaboração de um projeto contra-hegemônico. 6 NEGRO, A. L.; SILVA, S. (Orgs.) E. P. Thompson. As peculiaridades dos ingleses e outros artigos. Campinas: Editora da Unicamp, 2001, p. 208. UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... Nesse ínterim, as comemorações apresentadas pelo autor ― como o 1.º de Maio e os concursos de Rainha, por exemplo ― poderiam adquirir significados varia dos nesta verdadeira “luta de representações” do calendário operário (p. 399). O exercício contínuo de tentar captar ― a partir de uma apreciação cuidadosa das fontes ― as “raízes”, os valores costumeiros e a “invenção das tradições” da classe trabalhadora brasileira constitui, a nosso ver, uma das mais valiosas e inspiradoras contribuições de “A Reinvenção”. 6 NEGRO, A. L.; SILVA, S. (Orgs.) E. P. Thompson. As peculiaridades dos ingleses e outros artigos. Campinas: Editora da Unicamp, 2001, p. 208. Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265 UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... UM ANARQUISMO EXTRAORDINÁRIO: A LUTA DOS LIBERTÁRIOS... No capítulo 10, Leal procura problematizar os “bloqueios” enfrentados por aquilo que ele denominou de “novo bloco histórico” forjado por meio das lutas po pulares e de classe, ao longo deste período de mais de dez anos. Aqui, foi analisado como o PTB ― por sua desarticulação política e fragilidade eleitoral (p. 405) ― e o PCB ― pela missão imperiosa de cumprir “etapas históricas” (p. 432) ― obsta culizaram a possibilidade de se hegemonizarem como “representação política dos trabalhadores”. A concepção corporativa da política dos sindicalistas e os movi mentos, ora de “incorporação”, ora de repressão do sistema político partidário (p. 419), também são apontados como parte de um processo que impossibilitou a criação de um partido de trabalhadores. Cabe ressaltar que o conceito de “incor poração” das forças contra-hegemônicas utilizado pelo autor encontra-se inserido em um campo contínuo de correlação de forças. Resultado de sua tese de doutorado, defendida pelo Departamento de His tória da USP, o livro de Murilo Leal consiste em leitura obrigatória para os estudio sos da História Social do Trabalho. Por vezes, o “potencial contra-hegemônico da aliança interclassista” narrada pelo autor parece, de fato, ganhar “ares épicos”. Porém, a inegável qualidade da obra, somada às possibilidades analíticas suscita das pelo autor a partir da derrubada de “antigos mitos” e de novas matrizes inter pretativas, certamente contribuem para “oxigenar” antigos debates acadêmicos, suscitando novas polêmicas. Recebida em 4/11/2013 Aprovada em 6/12/2013 Recebida em 4/11/2013 Aprovada em 6/12/2013 265 265 Revista Mundos do Trabalho \| vol. 5 \| n. 10 \| julho-dezembro de 2013\| p. 261-265
https://openalex.org/W4220673083	https://zenodo.org/records/6374638/files/142-%20LV%20OK_130-140.pdf	Portuguese	null	Dificuldades de aprendizagem no Ensino Médio Profissional: a formação docente como possibilidade de intervenção	Zenodo (CERN European Organization for Nuclear Research)	2,022	cc-by	2,625	Brazilian Journal of High School doi: Electronic ISSN: 2595-816X Revista Brasileira do Ensino Médio (2022) \| vol. 5 \| 130-140 Brazilian Journal of High School doi: Electronic ISSN: 2595-816X Revista Brasileira do Ensino Médio (2022) \| vol. 5 \| 130-140 Revista Brasileira do Ensino Médio (2022) \| vol. 5 \| 130-140 Brazilian Journal of High School doi: Electronic ISSN: 2595-816X “O ENSINO MÉDIO INTEGRADO EM PERSPECTIVA” \| 2022 “O ENSINO MÉDIO INTEGRADO EM PERSPECTIVA” \| 2022 DOSSIÊ TEMÁTICO https://phprbraem.com.br/ojs/index.php/rbraem RESUMO Revista Brasileira do Ensino Médio 2022, Ipojuca (PE), Brasil https://phprbraem.com.br/ojs/index.php/rbraem \| 131/140 Dossiê: “O Ensino Médio Integrado em perspectiva” \| 2022 INTRODUÇÃO Rocha, R.; Caetano, M. R. / Dificuldades de Aprendizagem no Ensino Médio Profissional... 132/140 \| 132/140 \| A educação como percurso histórico inerente ao ser humano Antes de se pensar na educação enquanto prática social, há que se refletir sobre a capacidade humana que a possibilita: a aprendizagem. Faz-se necessária tal reflexão na medida em que a primeira parece ser uma sistematização de formas, normas, técnicas e práticas para que a segunda ocorra, a fim de se cumprirem objetivos práticos a partir de sua efetivação. Dito de outro modo, a educação tem como objetivo canalizar esta capacidade de aprender para fins específicos que, necessariamente terão impacto sobre as relações sociais. Assim, enquanto a aprendizagem, do ponto de vista biológico, é definida por Rotta (2016, p.04) como um processo que acontece no sistema nervoso central onde ocorrem modificações mais ou menos permanentes “[...] que se traduzem por uma modificação funcional ou conductual, permitindo uma melhor adaptação do indivíduo ao seu meio como resposta a uma solicitação interna ou externa [...]”, Paro (2014) a compreende como uma atualização histórico-cultural, tendo como objeto a cultura na sua integralidade, apontando claramente para o seu sentido social.No entanto, faz-se necessário compreender que tais entendimentos não se dão de forma isolada ao longo da construção da história da humanidade. Não é difícil deduzir que a capacidade de aprender, embora seja inerente ao ser humano, é individual e particular, enquanto que a educação se coloca como forma de Dossiê: “O Ensino Médio Integrado em perspectiva” \| 2022 \| 133/140 \| 133/140 Educação diante das diferenças Rocha, R.; Caetano, M. R. / Dificuldades de Aprendizagem no Ensino Médio Profissional... 134/140 \| As dificuldades de aprendizagem como parte integrante do processo educacional formal Antes de se falar sobre dificuldades de aprendizagem, faz-se necessário falar sobre a aprendizagem em seus diferentes significados. Sob o ponto de vista biológico, individual, tem-se que aprendizagem corresponde a alterações mais ou menos permanentes no sistema nervoso central, onde o “[...] ato de aprender é um ato de plasticidade cerebral, modulado por fatores intrínsecos (genéticos) e extrínsecos (experiência) ” (ROTTA, 2016, p. 97). A aprendizagem, porém, é um processo mais complexo que envolve diferentes dimensões da vida humana. Autores como Mosquera (2008), Paín (1992) e Vygotsky (2007) destacam a dimensão social da construção da aprendizagem, onde esta se insere em todos os momentos da vida do sujeito, processo contínuo com dimensões históricas e culturais. Vygotsky (2007, p. 32) aponta para a capacidade humana de desenvolver-se para além das “[...] funções psicológicas impostas pela natureza, evoluindo para uma organização nova, culturalmente elaborada [...], ” (VYGOTSKY,2007, p. 58). Embora seja inerente a todos os seres humanos, a aprendizagem não se dá de forma uniforme, ocorrendo diferenças de indivíduo para indivíduo pelos mais diferentes motivos. Conforme Rotta (2016, p.97) as dificuldades para a aprendizagem podem ser definidas como “[...] um termo genérico que abrange um grupo heterogêneo de problemas capazes de alterar as possibilidades de a criança aprender independentemente de suas condições neurológicas para fazê-lo”. Alícia Fernández (1990) considera que no desenvolvimento das dificuldades de aprendizagem podem intervir fatores ligados a aspectos socioeconômicos, educacionais, emocionais, intelectuais, orgânicos e corporais, propondo uma compreensão bastante abrangente da situação do não aprender. Um aspecto destacado por diferentes autores (SEABRA e colaboradores, 2014, FONSECA, 2015, OLWEILER, 2016), refere-se ao fato de que as dificuldades de aprendizagem, em muitos casos, ocorrem com alunos que apresentam funcionamento intelectual normal, no entanto seu rendimento em habilidades específicas, como leitura ou escrita, podem ficar abaixo do esperado. Segundo Fonseca (2018) o conceito de dificuldades de aprendizagem ainda hoje não é consensual, no entanto é amplamente reconhecida sua ocorrência e osproblemas que causa no ambiente escolar. Moojen e Costa (in ROTTA, 2016) dividem as dificuldades de aprendizagem em três categorias: aquelas transitórias, que desaparecem a partir de um esforço maior do estudante; as secundárias a uma condição que atinja o desenvolvimento como um todo e aquelas circunscritas a uma habilidade específica. Neste terceiro caso as autoras utilizam o termo transtorno de aprendizagem. As dificuldades de aprendizagem podem acompanhar o sujeito por toda a vida, assim, no ensino médio elas poderão se intensificar, tendo em vista as características deste nível de ensino bem como as particularidades de cada estudante. Tirado (2002) destaca a diversidade de interesse dos alunos, suas motivações e capacidades, aprendizagens pouco funcionais, desinteresse, história prévia permeada de suspensões, reprovações e repetência, nível de conhecimento abaixo do esperado para este nível de ensino, falta de comportamentos que favorecem a aprendizagem como pedir ajuda ou questionar quando tem dúvidascomo fatores que podem estar presentes ou agravarem o quadro. Sendo assim, as dificuldades de aprendizagem se colocam como mais uma situação no espaço escolar a ensejar práticas educativas inclusivas, visando garantir, também a estes alunos, o direito à educação. As dificuldades de aprendizagem como parte integrante do processo educacional formal Antes de se falar sobre dificuldades de aprendizagem, faz-se necessário falar sobre a aprendizagem em seus diferentes significados. Sob o ponto de vista biológico, individual, tem-se que aprendizagem corresponde a alterações mais ou menos permanentes no sistema nervoso central, onde o “[...] ato de aprender é um ato de plasticidade cerebral, modulado por fatores intrínsecos (genéticos) e extrínsecos (experiência) ” (ROTTA, 2016, p. 97). A aprendizagem, porém, é um processo mais complexo que envolve diferentes dimensões da vida humana. Autores como Mosquera (2008), Paín (1992) e Vygotsky (2007) destacam a dimensão social da construção da aprendizagem, onde esta se insere em todos os momentos da vida do sujeito, processo contínuo com dimensões históricas e culturais. Vygotsky (2007, p. 32) aponta para a capacidade humana de desenvolver-se para além das “[...] funções psicológicas impostas pela natureza, evoluindo para uma organização nova, culturalmente elaborada [...], Dossiê: “O Ensino Médio Integrado em perspectiva” \| 2022 \| 135/140 \| 135/140 todas as funções superiores originam-se das relações reais entre indivíduos humanos. ” (VYGOTSKY,2007, p. 58). Embora seja inerente a todos os seres humanos, a aprendizagem não se dá de forma uniforme, ocorrendo diferenças de indivíduo para indivíduo pelos mais diferentes motivos. Conforme Rotta (2016, p.97) as dificuldades para a aprendizagem podem ser definidas como “[...] um termo genérico que abrange um grupo heterogêneo de problemas capazes de alterar as possibilidades de a criança aprender independentemente de suas condições neurológicas para fazê-lo”. Alícia Fernández (1990) considera que no desenvolvimento das dificuldades de aprendizagem podem intervir fatores ligados a aspectos socioeconômicos, educacionais, emocionais, intelectuais, orgânicos e corporais, propondo uma compreensão bastante abrangente da situação do não aprender. Um aspecto destacado por diferentes autores (SEABRA e colaboradores, 2014, FONSECA, 2015, OLWEILER, 2016), refere-se ao fato de que as dificuldades de aprendizagem, em muitos casos, ocorrem com alunos que apresentam funcionamento intelectual normal, no entanto seu rendimento em habilidades específicas, como leitura ou escrita, podem ficar abaixo do esperado. Segundo Fonseca (2018) o conceito de dificuldades de aprendizagem ainda hoje não é consensual, no entanto é amplamente reconhecida sua ocorrência e osproblemas que causa no ambiente escolar. Moojen e Costa (in ROTTA, 2016) dividem as dificuldades de aprendizagem em três categorias: aquelas transitórias, que desaparecem a partir de um esforço maior do estudante; as secundárias a uma condição que atinja o desenvolvimento como um todo e aquelas circunscritas a uma habilidade específica. Neste terceiro caso as autoras utilizam o termo transtorno de aprendizagem. As dificuldades de aprendizagem podem acompanhar o sujeito por toda a vida, assim, no ensino médio elas poderão se intensificar, tendo em vista as características deste nível de ensino bem como as particularidades de cada estudante. Tirado (2002) destaca a diversidade de interesse dos alunos, suas motivações e capacidades, aprendizagens pouco funcionais, desinteresse, história prévia permeada de suspensões, reprovações e repetência, nível de conhecimento abaixo do esperado para este nível de ensino, falta de comportamentos que favorecem a aprendizagem como pedir ajuda ou questionar quando tem dúvidascomo fatores que podem estar presentes ou agravarem o quadro. Sendo assim, as dificuldades de aprendizagem se colocam como mais uma situação no espaço escolar a ensejar práticas educativas inclusivas, visando garantir, também a estes alunos, o direito à educação. todas as funções superiores originam-se das relações reais entre indivíduos humanos. Formação docente: aquele que ensina em constante aprendizado Um aspecto bastante peculiar na profissão docente é o fato de que todos os trabalhadores conhecem seu ofício desde tenra idade. Conforme Tardif e Raymond (2000, p. 224) “[...] o tempo de trabalho não se limita à duração da vida profissional, mas cobre também a existência pessoal dos professores, os quais, de um certo modo, aprenderam seu ofício antes de iniciá-lo. ” Tal situação, certamente influencia a forma como estes profissionais desenvolvem seu trabalho, levando para sua prática muito do que viram, sentiram e experimentaram ainda como alunos. Uma vez inseridos no contexto escolar, seguem construindo seu modo de fazer e agir enquanto professores, absorvendo regras e maneiras que nãosão necessariamente explicitadas ou formalizadas, mas que constituem o tecido que forma o seu saber sobre o seu ofício (TARDIF e RAYMOND, 2000). Trata-se do aspecto informal da formação do professor, adquirida com seus pares, abrangendo também atitudes e comportamentos, os quais acabam por constituir uma certa tradição, apoiada na cultura vivenciada na escola (idem). O outro nível da formação do professor é aquele formal, estruturado com objetivos e Rocha, R.; Caetano, M. R. / Dificuldades de Aprendizagem no Ensino Médio Profissional... 136/140 Procedimentos metodológicos Dossiê: “O Ensino Médio Integrado em perspectiva” \| 2022 \| 137/140 \| 137/140 Nos encontros presenciais foram propostas diferentes atividades onde os professores tiveram momentos para conversar, trocar experiências, refletir e discutir sobre os materiais disponibilizados no encontro não presencial. Considerações finais Rocha, R.; Caetano, M. R. / Dificuldades de Aprendizagem no Ensino Médio Profissional... 138/140 \| ANDRÉ, Marli. Formação de professores: a constituição de um campo de estudos. Revista Educação, Porto Alegre, v.33, nº3, p. 174-181, set/dez 2010 2. BRASIL. [Constituição (1988)]. Constituição da República Federativa do Brasil de 1988. Brasília, DF: Presidência da República, [2016]. Disponível em: http://www.planalto.gov.br/ccivil_03/Constituicao/ Constituiçao.htm. Acesso em 14 de fev de 2021.3. BRASIL. Decreto 5.154, de 23 de julho de 2004. Disponível em: http://www.planalto.gov.br/ccivil_03/_ato2004-2006/2004/decreto/d5154.htm. Acesso em 27 fev. de 2021. 4. BRASIL. Decreto nº 10.502.Política Nacional de Educação Especial: Equitativa, Inclusiva e com Aprendizado ao longo da Vida. Disponível em:http://www.planalto.gov.br/ccivil_03/_ato2019-2022/2020/decreto/D10502.htm. Acesso em 24 fev. de 2021. 5. BRASIL. Diretrizes Curriculares Nacionais Gerais da Educação Básica/ Ministério da Educação. Secretária de Educação. Básica. Diretoria de Currículos e Educação. Disponível em: portal.mec.gov.br/docman/julho-2013-pdf/13677-diretrizes-educacao-basica. Acesso em 26 dez. de 2020. 6. BRASIL. Estatutoda Criança e do Adolescente. Disponível em http://www.planalto.gov.br/ccivil_03/leis/l8069.htm. Acesso em 20 de fev. de 20217. BRASIL. Lei de Diretrizes e Bases da Educação Nacional. Disponível em http://www.planalto.gov.br/Ccivil_03/leis/L9394.htm. Acesso em 14 mar. de 2020. 8. BRASIL. INEP. Notas Estatísticas. Disponível em: www.portal.inep.gov.br/http://download.inep.gov.br/educacao_basica/censo_escolar/notas_estatisticas/2018/notas_estatisticas_censo_escolar_2018.pdf. Acesso em 25 mai. de 2020. 9. BRASIL. Nota Técnica nº 4/2014. Disponível em: http://portal.mec.gov.br/index.php?option= Referências \| 139/140 \| 139/140 Dossiê: “O Ensino Médio Integrado em perspectiva” \| 2022 com_docman&view=download&alias=1589 8nott04-secadi-dpee-23012014&category_slug=julho-2014-pdf&Itemid=30192. Acesso em 30 jun. de 2020 BRASIL. 10. MOOJEN, Sônia; COSTA, Adriana Corrêa. Semiologia psicopedagógica in ROTTA, Newra Tellechea; OHLWEILER, Lygia; RIESGO, Rudimar dos Santos. Transtornos de Aprendizagem: abordagem neurobiológica e multidisciplinar. 2. ed. Porto Alegre: Artmed, 2016. p.85-93. 20. BRASIL, Resolução CNE/CEB Nº 2/ 2001. Diretrizes Nacionais para a Educação Especial na Educação Básica. Disponível em: http://portal.mec.gov.br/arquivos/pdf/resolucao2.pdf Acesso em 14 jan. 2021 11. BRASIL. Resolução CNE/CEB nº 2/2013http:Disponível em http//portal.mec.gov.br/index.php?option=com_docman&view=download&alias =135 86-pceb002-13&Itemid=30192–. Acesso em 30 de dez 2020 12. MOSQUERA, Juan José Mouriño. Aprendizagem, significado e identidade em comunidades de prática. In ABRAHÃO, Maria Helena Menna Barreto (org.). Professores e alunos: aprendizagens significativas em comunidades de prática educativa. Porto Alegre: EDIPUCRS, 2008 21. FERNANDES, Alicia. A inteligência aprisionada: abordagem psicopedagógica clínica da criança e sua família. Tradução: Iara Rodrigues. Porto Alegre: Artes Médicas, 1990. 13. OLIVEIRA, Sandra de; WESCHENFELDER, Viviane Inês. Práticas in/exclusivas e os modos de ser professor (a) na contemporaneidade. In LOUREIRO, Carine Bueira; KLEIN, Rejane Ramos (orgs.), Inclusão e aprendizagem: contribuições para pensar as práticas pedagógicas. Curitiba: Appris, 2017. 22. FONSECA, Vitor da. Cognição, neuropsicologia e aprendizagem: abordagem neuropsicológica e psicopedagógica. Petrópolis, RJ: Vozes, 2015. 4ª reimpressão, 2018 14. FREIRE, Paulo. Pedagogia da autonomia:saberes necessários à prática educativa. Rio de Janeiro: Paz e Terra, 2013. 45ªed. 15. OHLWEILER, Lygia; RIESGO, Rudimar dos Santos. Transtornos de Aprendizagem: abordagem neurobiológica e multidisciplinar. 2. ed. Porto Alegre: Artmed, 2016. p.204-227. 23. FRIGOTTO, Gaudêncio. Concepções e mudanças no mundo do trabalho e o ensino médio. In: Gaudêncio Frigotto; Maria Ciavatta; Marise Ramos. (Org.). Ensino Médio Integrado: Concepção e contradição. 1ed.São Paulo: Cortez. 2005 16. PACHECO, José... [et al]. Caminhos para a inclusão: um guia para o aprimoramento da equipe escolar. Porto Alegre: Artmed, 2007. 24. FRIGOTTO, Gaudêncio. A polissemia da categoria trabalho e a batalha das ideias nas sociedades de classe. Revista Brasileira de Educação, v. 14, n. 40, jan. /abr. 2009 17. PAIN, Sara. Diagnóstico e tratamento dos problemas de aprendizagem. Trad. Ana Maria Netto Machado. Porto Alegre: Artes Médicas, 1992, 4ªed. 25. HATTGE, Morgana D., KLAUS, Viviane. Aimportância da pedagogia nos processos inclusivos. Revista Educação Especial. V. 27, nº49, p. 327-340, mai/ago.2014. Santa Maria. Disponível em http://www.ufsm.br/revistaeducacaoespecial18. PARO, Vitor Henrique. Educação como exercício do poder: crítica ao senso comum em educação. 3ª ed. São Paulo: Cortez, 2014. 26. PARO, Vitor Henrique. Referências Professor: artesão ou operário?São Paulo: Cortez. 2018 27. KUENZER,Acacia Zeneida. Palestra. In Formação de Professores para Educação Profissional e Tecnológica: Brasília, 26, 27 e 28 de setembro de 2006. –Brasília: Instituto Nacional de Estudos e Pesquisas Educacionais Anísio Teixeira, 2008. 304 p. –(Coleção Educação Superior em Debate; v. 8) 19. RAMOS, M. N. 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ROTTA, Newra T.; OHLWEILER, Lygia; RIESGO, Rudimar dos Santos (orgs.). Transtornos da aprendizagem: abordagem neurológica e multidisciplinar. 2ª ed. Poa: Artmed, 2016. 31. THOMA, Adriana da Silva; KRAEMER, Graciele Marjana. Inclusão no ensino superior: subjetivação e governo das diferenças in LOUREIRO, Carine Bueira; KLEIN, Rejane Ramos (orgs.). Inclusão eaprendizagem: contribuições para pensar as práticas pedagógicas. 1ª ed. Curitiba: Appris, 201737. SAVIANI, Dermeval. Trabalho e educação: fundamentos ontológicos e históricos. Revista Brasileira de educação. v. 12. n.34, jan/abr. 2007. 32. SEABRA, Alessandra Gotuzo, DIAS, Natália Martins, ESTANISLAU, Gustavo M. et al. Transtornos de aprendizagem. In Saúde mental na escola: o que os educadores devem saber. ESTANISLAU, Gustavo M., BRESSAN, Rodrigo Affonseca. Porto Alegre: Artmed, 2014. 33. VYGOTSKY, L. S. A formação social da mente: o desenvolvimento dos processos psicológicos superiores. 4ª ed. São Paulo: Martins Fontes, 2007. 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https://openalex.org/W4365812625	https://link.springer.com/content/pdf/10.1007/s11804-023-00321-1.pdf	English	null	Experimental Study on the Effect of Extreme Waves on a LNG Carrier	Journal of marine science and application/Journal of Marine Science and Application	2,023	cc-by	16,285	Marco Klein1,2, Shan Wang3, Günther Clauss4 and C. Guedes Soares3 Received: 27 November 2022 / Accepted: 18 January 2023 © The Author(s) 2023 Received: 27 November 2022 / Accepted: 18 January 2023 © The Author(s) 2023 Journal of Marine Science and Application (2023) 22: 52-74 https://doi.org/10.1007/s11804-023-00321-1 Journal of Marine Science and Application (2023) 22: 52-74 https://doi.org/10.1007/s11804-023-00321-1 RESEARCH ARTICLE Abstract This paper presents a comprehensive experimental study on the effect of extreme waves on a LNG carrier. The LNG carrier model was equipped with a variety of sensors to measure motions, green water height on deck as well as local and global loads. Experiments in transient wave packets provided the general performance in waves in terms of response amplitude operators and were accompanied by tests in regular waves with two different wave steepness. These tests allowed detailed insights into the nonlinear behavior of the vertical wave bending moment in steep waves showing that green water on deck can contribute to a decrease of vertical wave bending moment. Afterwards, systematic model tests in irregular waves were performed to provide the basis for statistical analysis. It is shown that the generalized extreme value distribution model is suitable for the estimation of the extreme peak values of motions and loads. Finally, model tests in tailored extreme wave sequences were conducted comparing the results with the statistical analysis. For this purpose, analytical breather solutions of the nonlinear Schrödinger equation were applied to generate tailored extreme waves of certain critical wave lengths in terms of ship response. Besides these design extreme waves, the LGN carrier was also investigated in the model scale reproduction of the real-world Draupner wave. By comparing the motions, vertical wave bending moment, green water column and slamming pressures it is concluded that the breather solutions are a powerful and efficient tool for the generation of design extreme waves of certain critical wave lengths for wave/structure investigations on different subjects. Keywords Extreme wave events; Wave-structure interaction; Draupner wave; Breather solutions 5 Technische Universität Berlin, Ocean Engineering Division, Salzufer 17-19, Berlin 10587, Germany * Marco Klein marco.klein@dlr.de 1 Introduction • A comprehensive experimental study on the impact of extreme waves on a LNG carrier is presented in terms of the ship motions, vertical bending moments (VBM), green water heights, and slamming loads. • A comprehensive experimental study on the impact of extreme waves on a LNG carrier is presented in terms of the ship motions, vertical bending moments (VBM), green water heights, and slamming loads. Extreme waves are characterized by significantly high wave amplitudes and short time duration, leading to large motions and loads on marine structures. The risk for ships encountering extreme sea states has been confirmed by the reported accidents in the last years with increasing fre‐ quency. A general overview of real sea extreme wave reg‐ istrations and reported accidents can be found in Kharif et al. (2008). The occurrence of hurricanes in the Gulf of Mexico has confirmed further that extreme sea states can be dangerous for marine structures. It is likely that the sig‐ nificance of severe sea state conditions for ship traffic will even grow in the future because of the expected increase in frequency and severity of extreme weather events asso‐ ciated with global warming. • The model tests in regular waves show that the green water on deck causes the decrease of VBM for wave lengthip length = 1.1. • The nonlinear behavior in higher, steeper waves can be investigated systematically in regular waves. • The statistical analysis for irregular wave tests shows that the GEV distribution model is suitable for the estimation of the extreme peak values of motions and loads. • The statistical analysis for Draupner extreme wave tests shows that the analytical breather solutions are good alternatives for the real work freak waves in the investigation of ship motions and loads. • The statistical analysis for Draupner extreme wave tests shows that the analytical breather solutions are good alternatives for the real work freak waves in the investigation of ship motions and loads. * Marco Klein marco.klein@dlr.de Even small wave heights with unfavorable wave lengths can cause a critical situation. et al. 2011, 2012). The code has been extended to include the body nonlinearities in the radiation and diffraction forces and then applied for predicting the vertical motions and loads for a cruise ship subject to large amplitude waves (Rajendran et al. 2015; Rajendran and Guedes Soares 2016). This time domain code has been coupled with a FE model in Rajendran et al. (2016) for investigating the flexible ver‐ tical response of container ships in high seas. Good results are obtained for low Froude numbers and head waves to strip theory codes; however, the accuracy decreases for other headings and higher speeds. The 3D Boundary Element Method (BEM) uses either a Wave Green’s Function (WGF) or a Rankine Source (RS) as a source potential to satisfy boundary conditions. Yasu‐ kawa (2002) have applied the 3D Rankine Panel method in the time domain to calculate ship motions and local pressure of several container ships with a large flare com‐ paring the results with model test and strip theory. Zakaria (2009) has used the linear 3D Panel method to predict the relative wave height of container ships in extreme seas. Guo et al. (2013) have studied ship response statistics in extreme seas using WASIM. Datta and Guedes Soares (2020) have analysed the hydroelastic response of a container vessel using a coupled BEM-FEM method. The application of RANS solver in simulating ship re‐ sponses in waves has been intensively investigated in recent years (Oberhagemann et al. 2012; Simonsen et al. 2013; Tezdogan et al. 2015;Jiao et al. 2021a, b). Most of them have been validated against experiments and show very prom‐ ising results. However, the numerical errors due to uncer‐ tainties are still a concern, and it is difficult to perform CFD simulations on many severe sea states due to high computational costs. Slamming and green water phenomenon often happens for various vessels in extreme seas. Clauss et al. (2012) have presented the systematic experimental study of an LNG car‐ rier and a chemical tanker in high, steep waves in the sea‐ keeping basin using breather solutions, showing that the impact of a extreme wave is severe and dangerous result‐ ing in large loads and huge amount of green water on deck. * Marco Klein marco.klein@dlr.de 1 German Aerospace Center (DLR), Institute of Maritime Energy Systems, Max-Planck-Straße 2, Geesthacht 21502, Germany Earlier investigations indicate that ship response may significantly increase in extraordinarily steep and/or large (extreme) waves and a proper prediction of responses due to extreme waves is important for ship safety in the extreme sea. There are only a few reported experimental studies with the ship responses in realistic extreme wave condi‐ tions. Stansberg and Karlsen (2001) have performed model tests with an FPSO in steep random waves showing that 2 Hamburg University of Technology, Offshore Dynamics Group, Schloßmühlendamm 30, Hamburg 21073, Germany 4 Centre for Marine Technology and Ocean Engineering (CENTEC), Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, Lisboa 1049-001, Portugal M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 53 the ship motions are an important parameter besides the encountering wave profile and indicating that the relative short and steep waves are more critical than longer waves, due to the phase lag of the pitch motion, in particular in combination with large pitch amplitudes. The vertical bend‐ ing moment (VBM) on FPSO due to extreme wave have been investigated by Clauss et al. (2004) and Guedes Soares et al. (2006) by comparing numerical simulations and model tests. The variation of the maximum bending moment with the position in space where the extreme wave was created has been investigated and found to have a small influence. Drummen et al. (2009) have compared the experimental and numerical responses of a modern container ship with a large bow flare in severe head seas. Denchfield et al. (2009) have calculated the symmetric motions and global wave-in‐ duced loads for a Leander class frigate in head irregular and extreme waves using a two-dimensional hydroelastic method comparing the rigid body motions with experiments. The investigation of Fonseca and Guedes Soares (2002) and Clauss et al. (2010) have shown for different types of ships that the geometry of the bow flare in combination with the wave steepness influence the global loads signifi‐ cantly, in particular the sagging loads. A segmented model of a RoRo vessel with a large bow flare has been tested in regular and two real-sea waves by Clauss et al. (2010), show‐ ing that critical loads and motions depend most notably on combinations of wave height, wave group sequences, crest steepness, encountering speed and the ships target position. * Marco Klein marco.klein@dlr.de Slamming impacts on a chemical tanker subjected to extreme waves have been investigated experimentally and numeri‐ cally in Wang and Guedes Soares (2016a, b). Slamming occurrence on the chemical tanker advancing in extreme waves modelled with the nonlinear Schrödinger equation has been studied by Wang et al. (2016). Wang et al. (2020) has studied the hydroelastic response of a river-sea-going containership. Green water loading on the hatch covers of bulk carriers in extreme weather conditions has been esti‐ mated by Vassalos et al. (2003) using empirical equations. The number of model tests investigating ship responses in extreme waves is still limited today while the existing nu‐ merical codes predicting ship behavior in these waves still need to be improved. To ensure the accuracy of numerical so‐ lutions, uncertainty quantification has been emphasized in re‐ cent years. The investigation of numerical errors due to discreti‐ zation of CFD solvers can be found in Huang et al. (2022) for wave loads and hydroelastic responses of a container ship, and in Wang et al. (2021) for slamming load predictions. The results of a benchmark study, organised by the ISSC-ITTC Joint Committee, on global linear wave loads on a container ship with forward speed has been presented in Parunov et al. (2022). The objective of the study was to assess the uncertainty in linear transfer functions due to different seakeeping codes and consequences on long-term extreme vertical wave bending moments. For this study, the well documented Flokstra’s container ship with accessible experimental data has been selected. The study has confirmed that large differences can be found between methods based on the same mathematical model and recommended the initiation of benchmark studies with different ship types to In addition to experimental analysis, numerical solutions based on potential flow are more frequently used. The time domain strip theory method (Fonseca and Guedes Soares 1998a, b) has been validated for predictions of responses to abnormal waves (Guedes Soares et al. 2008; Fonseca et al. 2.2.1 Pressure transducers The model was instrumented with pressure transducers at different positions on the hull: on the bow, on deck at the forecastle area and on the keel near the bow and at the stern. Figure 2 presents sketches of the pressure transducer locations at bow (Figure 2(a)) and stern (Figure 2(c)) as well as shows the installion of the pressure transducers at bow exemplary (Figure 2(b)). * Marco Klein marco.klein@dlr.de 2010), showing that the code overestimates the sagging moment peaks in very large and steep waves (Rajendran Journal of Marine Science and Application 54 Table 1　Main dimensions of the LNGC Parameters Length over all (LOA) Length between perpendicular (Lpp) Breadth Depth (moulded bulwark in the model) Draught Displacement Full scale 197.13 m 186.90 m 30.38 m 18.20 m 8.40 m 35 674.8 t Model scale 2.816 m 2.670 m 0.434 m 0.268 m 0.120 m 103.831 kg Table 1　Main dimensions of the LNGC provide the basis for the quantification of uncertainties. Within the EU project EXTREME SEAS, an extensive experimental study of the ship motions and wave-induced local and global loads of an LNG carrier in extreme seas has been conducted, however, only part of the experimental data have been made accessible for the public. To provide a general overview on the test setup, the experimental pro‐ gram and main findings as database for the benchmark study, this paper presents a comprehensive analysis of the impact of extreme waves on an LNG carrier, including the ship motions, VBM amidships, slamming, and green water impact loads of the model due to regular, irregular, and extreme waves. Main findings regarding motions and loads on the ship from previous studies will be summarized as well. the wave profile at the bow on the weather deck. Follow‐ ing, each individual measuring equipment is introduced in detail. 2 Model tests The model tests were conducted in the seakeeping basin of the Ocean Engineering Division of the Technical Uni‐ versity Berlin. The basin is 110 m long, with a measuring range of 90 m, the width is 8 m and the water depth is 1 m. On one side, an electrically driven wave generator is installed which can be used in piston-type as well as flap-type mode. The wave generator is fully computer controlled and a soft‐ ware is implemented which enables the generation of tran‐ sient wave packages, deterministic irregular sea states with defined characteristics as well as tailored critical wave sequences. On the opposite side, a wave-damping slope is installed to suppress wave reflections. The overall focus of the experiments lied on the pressure loads on the bow due to steep large waves. Therefore, a large area at the bow of the vessel was instrumented with pressure transducers to obtain conclusions regarding the lo‐ cal pressure distribution on the bow and the associated glob‐ al effects on the vbm (cf. Figure 2(a)). A detailed description of the position of the transducers as well as its measuring characteristics and uncertainties can be found in Appendix B. 2.2.2 Force transducers For the determination of the global loads, particularly the vertical wave bending moment, the ship model was di‐ vided into two segments which were connected with three force transducers. Two were mounted on deck, one on each side, the third one was mounted underneath the keel (cf. Figure 3). The force transducers registered the longitudinal forces during the model tests. Based on the measured forces and the given geometrical arrangement of the three force transducers, the resulting vertical wave bending moment and the longitudinal forces were obtained. The strain gauges featured a nominal load of 200 kg and a protection class of IP68 (100 h at 1 m water column). The LNGC model was made of fibreglass reinforced plastic (GRP) in model scale 1:70. The main dimensions are given in Table 1. An impression of the LNGC model is presented in Figure 1(a). The origin for all subsequent posi‐ tion specifications is shown in Figure 1(b). A detailed mass distribution of the investigated LNGC model is given in Table A1 in Appendix A. 2.2.3 Green water and relative wave gauges In addition to the two pressure sensors (No. 37 & No. 38) mounted on deck, two green water gauges (absolute, mea‐ surable green water height on deck is 0.185 m and 0.125 m) were installed at the same longitudinal positions. The posi‐ tions and dimensions are illustrated in Figure 4. Furthermore, a relative wave gauge (hrelative=0.75 m) was installed at the bow at the forward perpendicular. 2.2 Model equipment The model was equipped with several different sensors to measure global ship motions, global and local loads and Figure 1　Sketch of the investigated LNGC and the manufactured model Figure 1　Sketch of the investigated LNGC and the manufactured model M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 55 Figure 2　Overview on the pressure transducer locations Figure 2　Overview on the pressure transducer locations Figure 2 Overview on the pressure transducer locations Figure 2　Overview on the pressure transducer locations Figure 2　Overview on the pressure transducer locations Figure 3　Sideview of the mounted force transducers the model in position during the tests, the tracking system to measure the absolute motions as well as the wave gauges to measure the encountering waves. 2.3.2 Absolute motions The ship motions were recorded by an optical tracking system. The tracking system consists of a seven by ten meter frame that carries five infrared cameras which were shifted parallel to the moving ship model. The system enables high precision, contact-free motion tracking over large dis‐ tances by following the trajectories of infrared light emit‐ ting diodes mounted on the ship model. 2.3.1 Suspension system During the investigations, the model was fixed with an elastic suspension system using a triangular towing arrange‐ ment pulling the model without inducing a moment (cf. Figure 5(a)). The longitudinal motions were restricted by a spring in front of and a counter weight behind the model. With this arrangement, heave and pitch motions as well as the measured forces and moments remained unrestrained. Figure 3　Sideview of the mounted force transducers Figure 3　Sideview Figure 3 To evaluate the influence on the vbm, investigations were made in steep, regular waves with and without suspension system. Figure 5(b) compares the vbm at waterline level with and without suspension system. The investigation revealed that the influence is less than 2% and can be neglected. 3.1 Transient wave packet A powerful and efficient tool for the evaluation of the sea state behavior of ships and offshore structures is the transient wave packet technique which enables the determi‐ nation of the RAO efficiently in one test run (e.g., Clauss and Küuhnlein 1995, 1997; Clauss and Steinhagen 1999; Kühnlein et al. 2002; Clauss et al. 2010; Hennig 2005; Clauss et al. 2014; Klein et al. 2021). TWP's depict synthe‐ sized task-related wave spectra with tailored phase distri‐ bution. The phase distribution is adjusted in such a way that all components of the wave spectrum are in-phase superim‐ posed at the concentration point yielding a single wave peak. The shape and width of the spectrum can be adapted to the relevant frequency range of interest. Figure 7 presents the RAO’s determined by the TWP tech‐ nique. For comparative and evaluation purposes, the exper‐ imental results (blue curves) are compared with numerical results (red curves) obtained by the well established fre‐ quency domain analysis program WAMIT (Wave Analysis at Massachusetts Institute of Technology). WAMIT is a radiation/diffraction program for the evaluation of wave- structure interaction at zero speed in frequency domain (Newman 2018; WAMIT 1994; Lee 1995) being a widely 2.3.3 Wave gauge positions wave packages to obtain the Response Amplitude Operator (RAO) and the results were evaluated with tests in regular waves. Furthermore the model was investigated in irregu‐ lar sea states, critical wave sequences, and real world freak wave reproductions as well as in breather-type freak waves. In the following subsections, the different parts of the exper‐ imental program are presented. The number of wave probes as well as they positions were varying according to the objective of the test cam‐ paigns. Altogether, the LNGC was investigated at stationary conditions and at forward speed. At stationary conditions, one objective lied on a detailed detection of the wave evo‐ lution along the ship, particularly at the forward perpendic‐ ular. Figure 6 shows the sketch of the wave gauge posi‐ tions for the model test at stationary conditions. The inves‐ tigations were performed at a target location at 45.9 m, starting at the idle state of the wave board. Target point was the forward perpendicular of the LNGC. A total amount of ten wave gauges were installed about the target location. The first one is 6 m in front of it at xt1=39.9 m. A cluster of seven gauges, separated in 0.1 m intervals, was arranged around the target location to investigate the spatial evolution of the waves at the bow. Furthermore two gauges, one at each position, were installed amidships and at the aft perpendic‐ ular. For the cruising ship, this set-up was reduced to three wave gauges-one 6 m in front of the forward perpendicular (gauge No. 1 in Figure 6), one at forward perpendicular (gauge No. 5 in Figure 6) and one at amidships (gauge No. 9 in Figure 6) regarding the idle state of the model. 2.3 Test set-up The test set-up included the suspension system to hold Figure 4　Positions and dimensions of the relative wave gauge and the green water gauges Figure 4　Positions and dimensions of the relative wave gauge and the green water gauges Journal of Marine Science and Application 56 85 vbmwl (N·m) 100 50 −50 −100 0 90 (a) Top view on the test set-up (b) Comparison between vbm at waterline level measured with and without suspension system 95 100 105 110 t (s) 115 120 125 130 Free floating ship Suspension system Figure 5 Test set up and evaluation of the suspension system (b) Comparison between vbm at waterline level measured with and without suspension system t (s) Figure 5　Test set-up and evaluation of the suspension system 3.2 High and steep regular waves The investigated discrete regular wave lengths (Lw) were chosen in order to cover the complete range of interest in frequency domain-from Lw/Lpp=0.6 to Lw/Lpp=2.2. With regard to the influence of the wave steepness on the vbm, the test program was divided into two parts. Each part comprised the same wave lengths with varying wave steepness. The wave height and steepness was selected in such a way to obtain wave profiles with different crest/trough asymme‐ tries and to evaluate the influence of different wave profiles (asymmetries) on the vbm. Figure 8 illustrates the regions of validity of various grav‐ ity water wave theories with regard to the wave profile. The relative water depth is plotted on the abscissa and the relative wave height on the ordinate. It is shown that for small amplitude waves, the linear wave theory can be applied, whereas for increasing wave heights (and hence increasing crest/trough asymmetries) the Stokes II and Stokes III theo‐ ries become valid (the relative water depth of the investigated wave lengths is between 0.02≤d/(g·T 2)≤0.1 (cf. Figure 8). Figure 10 presents the green water column height on deck of the LNGC for the different wave length. The installed green water sensors are capable to detect continuous water volumes, but the green water on deck behaves like a spill‐ ing breaking wave with white cap. This results in a scatter of the detected green water column heights (indicated in Figure 10 via the associated variance). However, it is clear identifiable that the interval of the highest green water col‐ umns is within the observed hollow interval regarding the vbm. This shows that the vbm shrunk proportionally to the wave height due to the fact that some of the energy of the increased wave crest spills over the deck in terms of green water instead of acting on the bow resulting in a smaller vbm to wave height ratio. Furthermore, additional effects due to the influence of the shipping of water on deck have to be taken into account such as affected ship motions (Fon‐ seca and Guedes Soares 2005) as well as a counteracting moment due to the green water mass on deck (Rajendran et al. 2011; Clauss and Klein 2016). The first part of the experimental program included reg‐ ular waves with moderate amplitudes. 3 Experimental program and results The experimental program included investigations in fre‐ quency and time domain. One objective of the model tests was the determination of the response in terms of transfer functions. Therefore the model was investigated in transient Wave gauges 1-10 Wave-board 10 d=1 ma.pp. f.pp. s=0.1 m s s s s s s Lpp/2 9 8 7 6 5 4 3 2 1 48.57 m 47.235 m 45.9 m39.9 m Figure 6　Sketch of the wave gauge positions Wave-board Figure 6　Sketch of the wave gauge positions M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 57 technique. The sagging (red crosses) and hogging moments (red circles) are additionally registered to evaluate the asym‐ metry of the measured bending moment compared to the wave steepness. Figure 9(a) shows the result for wave pro‐ files in the Stokes II domain (WH2) and Figure 9(b) for wave profiles in the Stokes III domain (WH3). accepted, validated numerical tool for hydrodynamic anal‐ ysis and proven to be suitable for a multitude of applica‐ tions. Figure 7(a) presents the result for heave motion, Figure 7(b) for pitch motion and Figure 7(c) for the vbm. The good agreement between the experimental and numeri‐ cal results is evident at first glance. The result of the RAO’s determined in regular waves (\|sagging moment\| + hogging moment - red dashed curves) and in the transient wave packet (black curves) are in good agreement. With regard to the asymmetry of the hogging and sagging moments, it is clearly identifiable that the asym‐ metry between sagging and hogging increases with increas‐ ing wave height which is in accordance to previous investi‐ gations (Watanabe et al. 1989; Guedes Soares and Schellin 1998; Fonseca and Guedes Soares 2005; Clauss and Klein 2016). The trend of the vbm, in particular regarding the results in the WH3 regular waves, shows a special feature - a hollow at the interval where the maximum of the RAO would be expected for the LNG Carrier. This does not denote that the vbm shrunk but that the vbm shrunk pro‐ portionally to the wave height. It is noticeable that this occurs around Lw/Lpp = 1.1 and can be explained with the effect of green water on deck. 3.2 High and steep regular waves These se states, each about 30 min real time, were generated in group of up to six sea state sequences (phase seeds) per charac teristic sea state parameter set for long term statistical anal ysis and each row presents one set of the characteristic irregu 0.005 0.010 0.015 0.020 0.025 RAOWPT vbmtotal vbm2·hogging vbm2·sagging vbm 2 ρ·g·B·Lpp·H LNGC−Fn=0 WH2 - vertical bending moment at Lpp/2 LNGC−Fn=0 WH3 - vertical bending moment at Lpp/2 0.5 1.0 1.5 Lw/Lpp Lw/Lpp 2.0 (a) Results in regular waves of the Stokes II domain (b) Results in regular waves of the Stokes III domain 2.5 0.005 0.010 0.015 0.020 0.025 0.5 1.0 1.5 2.0 2.5 vbm 2 ρ·g·B·Lpp·H Figure 9　Comparison of the RAO for the vbm at Lpp/2 determined with the transient wave package technique (black curves) and the results i regular waves (red dashed curves) for the LNG Carrier. The sagging (red crosses) and hogging moments (red circles) are additionally registered Stokes Ⅳ or Ⅴ Stokes Ⅲ Stokes Ⅱ Non-breaking waves Linear wave theory (Airy) Ursell’s parameter Relative water depth 0.2 0.3 0.4 0.5 0.6 0.8 1.0 1.5 2.0 3.0 5.0 1.2 Cnoidal wave theory 0.000 4 0.001 0.002 0.004 0.006 0.01 0.02 0.04 0.06 0.1 0.2 0.3 0.4 stream function wave theory limit solitary wave Miche formula [Eq.(3.7)] limit deepwater waves breaking criterion HB L =0.142 HB 4 H= HL2 d3 =500 HL2 d3 =26 HB d =0.78 HB L =0.142 tanh(kd) d gT 2 LAiry 2π (k= ) kd πH L = tanh3 kd 3-tanh2 kd [Eq.(3.57)] Solitary wave theory relative wave height H gT 2 0.04 0.02 0.01 0.008 0.006 0.004 0.002 0.001 0.000 8 0.000 6 0.000 4 0.000 2 0.000 1 0.000 08 0.000 06 0.000 04 0.000 03 Shallow water -WH2 -WH3 Transitional water Deepwater Figure 8　Breaking wave height and regions of validity of various gravity water waves’ theories (Clauss et al. 3.2 High and steep regular waves The sagging (red crosses) and hogging moments (red circles) are additionally registered 0.005 0.010 0.015 0.020 0.025 RAOWPT vbmtotal vbm2·hogging vbm2·sagging LNGC−Fn=0 WH2 - vertical bending moment at Lpp/2 0.5 1.0 1.5 Lw/Lpp 2.0 2.5 vbm 2 ρ·g·B·Lpp·H vbm 2 ρ·g·B·Lpp·H LNGC−Fn=0 WH3 - vertical bending moment at Lpp/2 Lw/Lpp (b) R l i l f h S k III d i 0.005 0.010 0.015 0.020 0.025 0.5 1.0 1.5 2.0 2.5 (a) Results in regular waves of the Stokes II domain (b) Results in regular waves of the Stokes III domain Figure 9　Comparison of the RAO for the vbm at Lpp/2 determined with the transient wave package technique (black curves) and the results in regular waves (red dashed curves) for the LNG Carrier. The sagging (red crosses) and hogging moments (red circles) are additionally registered Figure 9　Comparison of the RAO for the vbm at Lpp/2 determined with the transient wave package technique (b regular waves (red dashed curves) for the LNG Carrier. The sagging (red crosses) and hogging moments (red circle 3.2 High and steep regular waves 1992) 58 58 Journal of Marine Science and Application Stokes Ⅳ or Ⅴ Stokes Ⅲ Stokes Ⅱ Non-breaking waves Linear wave theory (Airy) Ursell’s parameter Relative water depth 0.2 0.3 0.4 0.5 0.6 0.8 1.0 1.5 2.0 3.0 5.0 1.2 Cnoidal wave theory 0.000 4 0.001 0.002 0.004 0.006 0.01 0.02 0.04 0.06 0.1 0.2 0.3 0.4 stream function wave theory limit solitary wave Miche formula [Eq.(3.7)] limit deepwater waves breaking criterion HB L =0.142 HB 4 H= HL2 d3 =500 HL2 d3 =26 HB d =0.78 HB L =0.142 tanh(kd) d gT 2 LAiry 2π (k= ) kd πH L = tanh3 kd 3-tanh2 kd [Eq.(3.57)] Solitary wave theory relative wave height H gT 2 0.04 0.02 0.01 0.008 0.006 0.004 0.002 0.001 0.000 8 0.000 6 0.000 4 0.000 2 0.000 1 0.000 08 0.000 06 0.000 04 0.000 03 Shallow water -WH2 -WH3 Transitional water Deepwater Figure 8　Breaking wave height and regions of validity of various gravity water waves’ theories (Clauss et al. 1992) Figure 8　Breaking wave height and regions of validity of various gravity water waves’ theories (Clauss et al. 1992) 0.005 0.010 0.015 0.020 0.025 RAOWPT vbmtotal vbm2·hogging vbm2·sagging vbm 2 ρ·g·B·Lpp·H LNGC−Fn=0 WH2 - vertical bending moment at Lpp/2 LNGC−Fn=0 WH3 - vertical bending moment at Lpp/2 0.5 1.0 1.5 Lw/Lpp Lw/Lpp 2.0 (a) Results in regular waves of the Stokes II domain (b) Results in regular waves of the Stokes III domain 2.5 0.005 0.010 0.015 0.020 0.025 0.5 1.0 1.5 2.0 2.5 vbm 2 ρ·g·B·Lpp·H Figure 9　Comparison of the RAO for the vbm at Lpp/2 determined with the transient wave package technique (black curves) and the results in regular waves (red dashed curves) for the LNG Carrier. 3.2 High and steep regular waves The relative wave heights of the investigated waves were between 0.002 ≤H/ (g·T 2) ≤0.004 - where the wave profile was within the Stokes II domain (cf. Figure 8 - red dots). During the sec‐ ond part of the experimental program, the same (regular) wave lengths were generated with increased relative wave heights and thus the relative wave heights were between 0.01≤H/(g·T2)≤0.012 - where the wave profile was within the Stokes III domain (cf. Figure 8 - blue dots). Figure 9 presents the results of the model tests in regu‐ lar waves-the respective RAOs (\|sagging moment\| + hog‐ ging moment-red dashed curves) are compared to the RAO (black curves) determined by the transient wave packet vbm (Nm/m) ×108 0.8 0.8 1.0 1.0 1.2 1.2 1.4 1.6 1.8 2.0 Model test WAMIT 0.6 0.6 0.4 0.4 0.2 0.2 0 ω (rad/s) 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.5 ω (rad/s) 1.0 1.5 s3 (m/m) s5 (°/m) vbm (Nm/m) 0 0.9 ×108 0.8 0.8 0.8 1.0 1.0 1.2 1.2 1.4 1.6 1.8 2.0 Model test WAMIT 0.7 0.6 0.6 0.6 0.5 0.4 0.4 0.4 0.3 0.2 0.2 0.2 0.1 0 0 0.5 ω (rad/s) ω (rad/s) 1.0 1.5 Figure 7　Comparison of measured (blue curves) and calculated RAOs (red curves) for the LNGC s5 ( /m) 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 0.5 ω (rad/s) 1.0 1.5 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.5 ω (rad/s) 1.0 1.5 s3 (m/m) 0 Figure 7　Comparison of measured (blue curves) and calculated RAOs (red curves) for the LNGC Journal of Marine Science and Applicatio 3.3 Irregular sea states with random phases Several irregular sea states were investigated representing design storms according to existing probabilistic methods. The investigated characteristic sea state parameters Tp, Hs and γ are given in Table C3 in Appendix C. M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 59 0.5 1.0 1.5 2.0 0 0.5 1.0 1.5 2.0 2.5 3.0 LNGC−Fn=0 WH3-green water column at forward perpendicular Lw/Lpp ζgreen water (m) Figure 10　Green water column height on deck of the LNGC at the forward perpendicular 0.5 1.0 1.5 2.0 0 0.5 1.0 1.5 2.0 2.5 3.0 LNGC−Fn=0 WH3-green water column at forward perpendicular Lw/Lpp ζgreen water (m) 3 0 LNGC−Fn=0 WH3-green water column at forward perpendicular IRREGULAR20 (cf. Table C3) at Fn=0 are presented. The focus lies on the statistical analysis of the excceedance prob‐ ability of the peaks of global motions and loads. For com‐ parative purposes, the experimental results are compared to numerical simulations. Therefore, a strip theory based partially nonlinear time domain code was applied (Rajen‐ dran et al. 2015). Figure 11 compares the time series of the numerical solution and the measured incident waves, heave and pitch motions, and the vbm at amidships (Fn=0 and sea state IRREGULAR20). The wave profile recorded at amidships (cf. No. 9 in Figure 6) were directly converted to the input data for the numerical simulation. As seen in Figure 11(a), the numerical wave sequence is in perfectly good agree‐ ment with the measurements. However, as the wave pro‐ file in space domain is based on linear transformation of the registration amidships, the wave profile along the ship in space domain will differ from the experiments. In addi‐ tion and in contrast to the experiments, the ship’s position in space is fixed. Reading the motions (Figure 11(b) and 11(c)) and loads (Figure 11(d)), the agreement between numerical and experimental values are satisfactory in general, though the numerical method overpredicts the peaks slightly. Figure 10　Green water column height on deck of the LNGC at the forward perpendicular lar sea state parameters. The phases were randomly gener‐ ated and differ within each set of parameters but are con‐ stantly plugged for each set. For example, parameter set No. 1 (see Table C3), phase seed #1 had another random phase distribution than phase seed #2, #3 and so on of the same set, but the same phase seed like set No. 2, phase seed #1, set No. 3, phase seed #1 and so on. Altogether, 45 irregular sea states for test runs at stationary conditions (Fn=0) as well as at forward speed (Fn=0.07). 3.3 Irregular sea states with random phases and γ are given in Table C3 in Appendix C. These sea states, each about 30 min real time, were generated in groups of up to six sea state sequences (phase seeds) per charac‐ teristic sea state parameter set for long term statistical anal‐ ysis and each row presents one set of the characteristic irregu‐ Several irregular sea states were investigated representing design storms according to existing probabilistic methods. The investigated characteristic sea state parameters Tp, Hs M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier Figure 12 shows the comparison of the exceedance prob‐ ability of the peaks for IRREGULAR20 phase seed #1. In the following, the experimental results for sea state Figure 11 Comparison of the numerical prediction of the time series of the wave surface elevation at amidships heave pitch and vbm with Figure 11　Comparison of the numerical prediction of the time series of the wave surface elevation at amidships, heave, pitch and vbm with experiments in the sea state IRREGULAR20 phase seed #1 Figure 11　Comparison of the numerical prediction of the time series of the wave surface elevation at amidships, heave, pitch and vbm with experiments in the sea state IRREGULAR20 phase seed #1 Journal of Marine Science and Application 60 For the experimental data, negative heave peaks are larger than the positive ones for the measured responses except the tails, while the positive pitch peaks are higher than the negative ones. The numerical heave peaks follow similar trends for all the results; however, they are all higher than the experimental peaks at the same probability. small (below 3.2 %) for the Fn=0 case. For the case with a forward speed (Fn=0.07), all uncertainties are below 2.2% (cf. Wang and Guedes Soares 2022b). Compared with the random uncertainty of the slamming peaks, the experimen‐ tal errors of motions and vbm are quite small. A detailed analysis and discussion can be found in Wang and Guedes Soares (2022b). Taking all five phase seeds of IRREGULAR20 into account gives the exceedance probability of the peaks pre‐ sented in Figure 13. For the heave motion, the positive and negative values are very close when the exceedance proba‐ bility is high, but the difference becomes larger when the exceedance probability is getting smaller. A similar obser‐ vation is found for pitch motion. Comparing the values from the five repeated tests, the exceedance probabilities for the peaks agree quite well for small values, but more deviations are found for larger values. It is expected that the uncertainty in extreme value is higher. M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 61 Figure 13　Exceedance probability of peaks for the motions and vbm in sea state IRREGULAR20 all five phase seeds Figure 13　Exceedance probability of peaks for the motions and vbm in sea state IRREGULAR20 all five phase seed 2008), was generated in the seakeeping basin. Exact solu‐ tions of the non-linear Schrödinger (NLS) equation-the so called breather solutions-were applied for the generation of design extreme wave groups. Breather solutions are characterized by quasi-monochromatic waves being slightly perturbed in time or space. Due to modulational instability, this perturbation increases during evolution resulting in large amplifications of the initial amplitude. Altogether, three types of breather solutions, namely the Kusnetzov-Ma breather (Kuznetsov 1977; Ma 1979), the Akhmediev breath‐ er (Akhmediev et al. 1985; Akhmediev and Korneev 1986; Akhmediev et al. 1987) and the Peregrine breather (Peregrine 1983), were utilized in order to evaluate their applicability for task related research questions on extreme wave impact. 2008), was generated in the seakeeping basin. Exact solu‐ tions of the non-linear Schrödinger (NLS) equation-the so called breather solutions-were applied for the generation of design extreme wave groups. Breather solutions are characterized by quasi-monochromatic waves being slightly perturbed in time or space. Due to modulational instability, this perturbation increases during evolution resulting in large amplifications of the initial amplitude. Altogether, three types of breather solutions, namely the Kusnetzov-Ma breather (Kuznetsov 1977; Ma 1979), the Akhmediev breath‐ er (Akhmediev et al. 1985; Akhmediev and Korneev 1986; Akhmediev et al. 1987) and the Peregrine breather (Peregrine 1983), were utilized in order to evaluate their applicability for task related research questions on extreme wave impact. numerically that the Peregrine breather is applicable for modeling extreme waves in realistic oceanic conditions by embedding the Peregrine breather in complex sea states based on JONSWAP spectra. In this context, Klein et al. (2016) have shown experimentally that Peregrine breathers can be embedded into irregular sea state to tailored extreme wave sequences and that such a design extreme wave features simi‐ lar dynamics compared to the real world Draupner wave. Following, the experiments with the three breather solu‐ tions are presented and compared with the results in the Draupner wave. Each solution was used to generate a set of high, steep single waves of different discrete carrier wave lengths to cover the range of interest in frequency domain regarding the vbm. M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier The initial steepness at the wave board were defined in such a way to ensure that the high single wave at target location is as high as physically possible for the defined wave lengths. The application of breather solutions for model tests is beneficial due to several reasons. One aspect relates to the simple and straightforward applicability as design extreme wave: “The major benefits are the potential to generate abnormal waves of certain frequency up to the physically possible wave height, the symmetrical shape of the extreme wave and the availability of an analytical solution”(Klein et al. 2016), which enables a fast and simple generation of the wave maker control signal. Another aspect relates to the fact that breather solutions, particularly the Peregrine breather, are being considered as special prototype of extreme waves (Shrira and Geogjaev 2010). Wang et al. (2018) have shown 3.4 Extreme wave events Investigations on extreme wave events are indispensable for a complete evaluation of a ship. Besides the statistical determination of extreme responses based on investigations in irregular sea state with predefined design wave spectra (and random phase distributions), investigations in deter‐ ministic extreme wave sequence enable the deterministic evaluation of critical wave sequences and extreme responses. Particularly, experiments in deterministic extreme wave events allows the detailed analysis of the cause-reaction chain for specific research questions and the evaluation if such wave events are critical for the investigated structure which cannot covered by statistical analysis. To study the peak values of the ship motions and loads subjected to extreme waves, the generalized extreme value (GEV) distribution model was applied on the experimental data. The statistical analysis showed that the GEV distribu‐ tion model is suitable for the estimation of the extreme peak values. Figure 14 presents diagnostic plots of good‐ ness of fit for generalized extreme value distribution of the peak values from the experiments. The random uncertain‐ ties of the experimental data for the motions and vbm are The experiments on extreme wave events comprised inves‐ tigations based on real world measurements as well as spe‐ cific design extreme wave groups. As real world reproduc‐ tion, the famous Draupner wave, also known as “New Year Wave” (Slunyaev et al. 2005; Cherneva and Guedes Soares Figure 12　Comparison of the exceedance probability of the peaks for the motions and vbm in sea state IRREGULAR20 phase seed #1 Figure 12　Comparison of the exceedance probability of the peaks for the motions and vbm in sea state IRREGU omparison of the exceedance probability of the peaks for the motions and vbm in sea state IRREGULAR20 phase seed # Figure 12　Comparison of the exceedance probability of the peaks for the motions and vbm in sea state IRREGULAR20 phase seed #1 M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 3.4.1 Breather-type extreme waves The three breather solutions applied in the experiments represent exact solutions of the NLS equation which reads ∂A ∂x + 1 Cg ∂A ∂t + i(α' ∂2A ∂t2 + β'\|A\|2A) = 0 (1) (1) for the wave group evolution in space of a time series, with 62 Journal of Marine Science and Application Figure 14　Diagnostic plots of goodness of fit for generalized extreme value (GEV) distribution of the peak values from the experiments Figure 14　Diagnostic plots of goodness of fit for generalized extreme value (GEV) distribution of the peak values from the experiments α' = 1 8 ωc kc 2 α Cg 3 (2) β' = 1 2 ωckc 2 β Cg (3) α' = 1 8 ωc kc 2 α Cg 3 (2) and β' = 1 2 ωckc 2 β Cg (3) The carrier wave number is represented by kc, the corre‐ sponding angular frequency by ωc, the group velocity by Cg and the water depth by d. The correction for the group velocity in finite water depth is taken into account by the coefficient ν, (2) and (3) (3) ν = 1 + 2 kcd sinh (2kcd ) (6) (6) The coefficients α and β read for arbitrary depth (Serio et al. 2005) 3.4.1.1 The Akhmediev breather 3.4.1.1 The Akhmediev breather Consequently, this solution does not comply with the classical Benjamin-Feir instability. The corresponding analytical solution is The Kusnetzov-Ma breather (Kuznetsov 1977; Ma 1979) is periodic in space and decreases exponentially in time. Contrary to the Akhmediev breather, the modulation for the Kusnetzov-Ma breather is never small, i.e, the solution oscillates while propagating. Consequently, this solution does not comply with the classical Benjamin-Feir instability. The corresponding analytical solution is AB ( x, t ) = Ac ( x )( − 2 ϑ͂ 2cos(ηx ) + i 2 ῆ sin (ηx ) 2 cos(ηx ) − 2 + ϑ͂ 2 cosh (ϑt ) −1) (8) Figure 15 presents the results of the model tests at sta‐ tionary conditions (Fn=0) in breather-type extreme waves- from top, the shortest, to bottom the longest wave length. The LNGC was investigated in three different carrier wave length (Lw/Lpp=0.7 to Lw/Lpp=1.1) for each of the three breather solutions. In addition, the Peregrine breather was exclusively applied for the Lw/Lpp=1.3. The results of each carrier wave length comprises a set of three diagrams-the top diagram shows the surface elevation at the forward per‐ pendicular, the center diagram the vbm at amidships and the bottom diagram the green water column height on deck at the forward perpendicular. (8) with ϑ͂ = ϑ/ac α'/β', ῆ =ϑ͂ 2 + ϑ͂ 2 and η = β' a2 cῆ (Karjanto and van Groesen 2007). Three parameters have to be predefined in order to deter‐ mine the breather envelope-the plane-wave amplitude ac, the normalized modulation wave number ῆ as well as the carrier frequency ωc. 3.4.1.3 The Peregrine breather The Peregrine breather solution (Peregrine 1983), also known as rational solution, represents the limiting case of the time-periodic Kusnetzov-Ma breather and the space- periodic Akhmediev breather. It has the specific character‐ istic that it is neither temporally nor spatially periodic: it is a wave that “appears from nowhere and disappears without trace” (Akhmediev et al. 2009) being considered as special prototype of freak wave (Shrira and Geogjaev 2010). Its analytical form suitable for wave tank experiments is (Kar‐ janto and van Groesen 2007) The design vertical wave bending moments according to the IACS-Common Rules (IACS 2015) are additionally illustrated to evaluate the measured vbm, knowing that in the dimensioning process of the transverse section additional parameters have to be considered (e.g., γR, cs, deck open‐ ings, ...). 3.4.1.1 The Akhmediev breather 3.4.1.1 The Akhmediev breather The Akhmediev breather (Akhmediev et al. 1985; Akhme‐ diev and Korneev, 1986; Akhmediev et al. 1987) depicts the modulational instability process in space and is periodic in time, α =−ν2 + 2 + 8(kcd )2 cosh (2kcd ) sinh2 (2kcd ) (4) (4) β = cosh (4kcd ) + 8 −2tanh2 (kcd ) 8sinh4 (kcd ) − β = cosh (4kcd ) + 8 −2tanh2 (kcd ) 8sinh4 (kcd ) − (2cosh2 (kcd ) + 0.5ν )2 sinh2 (2kcd ) é ë êêê ù û úúú kcd tanh (kcd ) −( ν 2 )2 (5) , AB (x, t) = Ac (x)( ) ῦ 2cosh (σx ) −iσ͂ sinh (σx ) cosh (σx ) − 1 −1 2 ῦ 2 cos(υt ) −1 (7) (5) (7) M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 63 Lw/Lpp=0.7 to Lw/Lpp=1.3. The initial steepness at the wave board was defined in such a way that the high single wave at target location is as high as physically possible for the defined wave lengths. with ῦ = υ/ac α'/β', σ͂ =ῦ 2 + ῦ 2 and σ = β' a2 cσ͂ (Karjanto and van Groesen 2007). To arrive at the Akhmediev breather solution, three parameters have to be predefined - the plane-wave amplitude ac, the normalized modulation fre‐ quency ῦ as well as the carrier frequency ωc. 3.4.1.2 The Kusnetzov-Ma breather Based on the different solutions, the determined surface elevation at the wave board was divided by the RAO of the wave maker to obtain the control signal. The agree‐ ment between theoretical location of freak wave occur‐ rence (input) and the registrations/observations in the tank showed satisfying agreement, i.e., in most of the cases the actual freak wave event was nearby the given value. But due to the fact that wave-structure investigations implies the definition of target waves in time domain at precise tar‐ get locations, the investigated breather-type extreme waves presented here resulting from an adjustment of the first control signal to obtain the extreme wave as exact as possible at the target location, i.e., which took up to 3 iteration steps. The Kusnetzov-Ma breather (Kuznetsov 1977; Ma 1979) is periodic in space and decreases exponentially in time. Contrary to the Akhmediev breather, the modulation for the Kusnetzov-Ma breather is never small, i.e, the solution oscillates while propagating. 3.4.1.1 The Akhmediev breather The black dashed lines denote the design vertical wave bending moment. The ultimate design vertical wave bending moments, which can be calculated based on design vbm considering safety factors, are not displayed as they are outside the axis scaling (i.e., all data is below this threshold). AB ( x, t ) = Ac ( x )( 4α' (1 −i2β'a2 c x ) α' + α' (2β'a2 c x )2 + 2β'a2 ct2 −1) (9) The ordinates are equally scaled for all associated dia‐ grams to simplify the overall picture of the results. The dif‐ ferent results of the three breather solutions for the same carrier wave length are illustrated in the same set of dia‐ grams-the blue curves denote the results for the Peregrine breather, the red curves for the Akhmediev breather and the green curves the Ma-Kuznetsov breather. In general, only the plane-wave amplitude ac as well as the carrier frequency ωc (i.e., the initial steepness) needs to be predefined to fully determine the Peregrine solution in space and time. 3.4.1.4 Experimental results The overall picture of the presented results shows that the high single wave is clearly identifiable at the forward perpendicular-for all breathers and wave length to ship length ratios. Furthermore, it is obvious that the surface elevation of the different breathers with the same carrier wave lengths is quite identical as well as the associated responses, but the Akhmediev breather shows a slightly different wave propagation in front of the target wave. The height of the The LNGC was investigated in the three breather solu‐ tions introduced above in order to evaluate the applicability of each solution as well as to determine the impact of such high, steep single wave on the LNGC. At the beginning, the three solutions were used to generate a set of high, steep single waves of different discrete carrier wave lengths. 3.4.1.1 The Akhmediev breather The carrier wave lengths were chosen in order to cover the range of interest in frequency domain regarding the vbm-from 64 Journal of Marine Science and Application 15 10 ζ (t) (m) vbm (t) (kN·m) 5 0 −5 −0.5 −1.0 12 10 8 6 4 2 0 −1.5 −2.0 −10 20 1.5 2.0 1.0 0.50 Surface elevation at forward perpendicular ×106 Green water on deck at forward perpendicular Peregrine breather Akhmediev breather Ma-kuznetsov breather MWV 680 700 720 740 760 780 800 820 Vertical bending moment at Lpp/2 (a) Lw/Lpp=0.7 ζgreen water (t) (m) 15 10 5 0 −5 −0.5 −1.0 12 10 8 6 4 2 0 −1.5 −2.0 −10 20 1.5 2.0 1.0 0.50 Surface elevation at forward perpendicular ×106 Green water on deck at forward perpendicular 680 660 640 620 600 700 720 740 760 Vertical bending moment at Lpp/2 ζgreen water (t) (m) ζ (t) (m) vbm (t) (kN·m) (b) Lw/Lpp=0.9 15 10 5 0 −5 −0.5 −1.0 12 10 8 6 4 2 0 −1.5 −2.0 −10 20 1.5 2.0 1.0 0.50 Surface elevation at forward perpendicular ×106 Green water on deck at forward perpendicular 600 550 650 700 Vertical bending moment at Lpp/2 ζgreen water (t) (m) ζ (t) (m) vbm (t) (kN·m) (c) Lw/Lpp=1.1 15 10 5 0 −5 −0.5 −1.0 12 10 8 6 4 2 0 −1.5 −2.0 −10 20 1.5 2.0 1.0 0.50 Surface elevation at forward perpendicular ×106 Green water on deck at forward perpendicular 580 560 540 520 600 620 640 660 Vertical bending moment at Lpp/2 ζgreen water (t) (m) ζ (t) (m) vbm (t) (kN·m) (d) Lw/Lpp=1.3 of the model tests in breather-type extreme waves Figure 15　Results of the model tests in breather-type extreme waves M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 65 extreme waves is severe and a limiting case for the investi‐ gated ship. This is confirmed by the measured green water column heights at the forward perpendicular which reaches impressive values (cf. Figure 10 and Figure 15) - a 10 m high wave front had been detected at the forward perpen‐ dicular spilling over the deck which is as high as a single family home. In addition, very high slamming pressure were captured at the bow (pressure sensors No. 02 and No. 05) and stern (pressure sensors No. 31 and No. 3.4.1.1 The Akhmediev breather 32) of the model for the Lw/Lpp= 1.3 as seen in Figure 16. For all four re‐ corded pressure sensors, the maximum value occurred when the highest wave impacts the hull. In this Peregrine breath‐ er solution, the maximum pressure peaks are 1.07 bar, 1.28 bar, 1.01 bar, and 1.16 bar, respectively. For the Lw/Lpp= 1.1 cases, the Peregrine breather solution gives the peaks val‐ ues as 0.88 bar, 1.00 bar, 0.67 bar, and 0.87 bar, and the Ma-Kuznetsov breather solution provides the peaks as 0.95 bar, 0.99 bar, 0.71 bar, and 0.88 bar, while the Akhmediev breather predicts the peaks as 0.95 bar, 1.18 bar, 0.85 bar, and 1.03 bar. By using the GEV distribution, the most probable extreme values of the peaks on pressure sensors No. 02, No. 05 and No. 32, are 0.82 bar, 0.91 bar and 0.73 bar, for the case in sea state IRREGULAR20 with Hs=11.5 m and Tp=12 s. All the peaks values from the three breather solu‐ tions are higher than the extreme values in the irregular presented extreme waves increases with increasing carrier wave length independently of the used breather solution due to the fact that all presented waves are generated in such a way that the highest physically possible wave height is obtained at the target location, i.e., all presented waves are plunging breaker close to the target location. Analogue to the results in regular waves, the resulting vbm depends on the wave length and reaches maximum values for the identified critical wave lengths as a function of the wave height. But also the reached absolute values are outstanding due to the impact of the highest physically possible wave height, in contrast to the tests in regular waves with moderate wave steepness. The wave heights of the highest, steepest regular waves (WH3) are approximately 40% smaller compared to the breathers. Comparing the abso‐ lute vbm values show that the moments are at least 15% higher for hogging condition and 30% higher for sagging condition than measured in regular waves. The maximum vbm, in particular the sagging moment, reaches the design vertical wave bending moment regarding the LNG Carrier. The exceeding of the design vertical wave bending moment is not resulting in an inevitable structural failure of the ship at full scale as additional parameters have to be considered for the dimensioning of the transverse section. 3.4.1.1 The Akhmediev breather Nevertheless, it shows that the impact of the breather-type Figure 16　Slamming pressure at the bow and stern for Fn=0 in Peregrine Breather extreme wave for Lw/Lpp=1.3 Figure 16　Slamming pressure at the bow and stern for Fn=0 in Peregrine Breather extreme wave for Lw/Lpp=1.3 Journal of Marine Science and Application 66 case. In particular, the peaks of the the Peregrine breather Lw/Lpp= 1.3 case are 31%, 40% and 58% higher than those extreme values. resent abnormal wave events which happened on sea in con‐ trast to predefined breathers. Therefore, the results obtained in real world freak wave reproductions exclude all possi‐ bility of doubt regarding discussions on possible investi‐ gated unrealistic, artificial wave sequences. However, this result denotes that the breather solutions can be used to design extreme wave events for certain wave lengths up to the maximum possible wave height for the identification of critical wave sequences regarding wave length, height and steepness. Altogether, the results show that the breather solutions are a powerful tool for the generation of tailored extreme waves of certain critical wave lengths for investigations of limiting cases on different subjects, e.g., local and global loads, green water effects as well as air gap investigations. In this context, the Peregrine breather solution is particu‐ larly advantageous (Klein et al. 2016). To investigate the impact of the Draupner wave on the motions and loads of the ship, Figure 19 shows the exceed‐ ance probability of maxima and minima of the heave and pitch motion as well as the vbm amidships, together with the analytical distribution models by fitting these peaks. It is seen that all the positive peaks are higher than the nega‐ tive ones for the vbm. The positive peaks are larger than the negative ones at the tails for the distribution of heave and pitch motions. The GEV model fits better on the maxima and minima of the motions, while the Weibull model per‐ forms a bit better on the peaks of vbm. By using the GEV model, the most probable extreme values of heave motion are 5.31 m (positive) 6.60 m (negative), and of pitch mo‐ tion are 7.92° and 7.82°. With the Weibull model, the most probable extreme values of vbm are 1.40×109 kN in hog‐ ging and 1.63×109 kN in sagging. 3.4.2 Real world extreme wave reproduction To evaluate the model test results obtained using the breather solutions, the LNGC was also investigated in a real- world extreme wave reproduction - the famous Draupner wave. The Draupner extreme wave has been recorded dur‐ ing a storm on January 1, 1995 at the Draupner platform in the North Sea. A giant single wave (Hmax=25.63 m) with a crest height of ζc=18.5 m occurred in a surrounding sea state characterised by a significant wave height of Hs= 11.92 m (Hmax/Hs=2.15). The water depth at the location is d=70 m. Figure 17 shows the measurement in the wave tank in comparison to the original wave sequence recorded at the Draupner platform-see Clauss and Klein (2011) for details on the wave generation procedure. Figure 18 presents the results for the LNGC at station‐ ary conditions (Fn=0) in the Draupner wave. Figure 18(a) displays the surface elevation at the forward perpendicular, Figure 18(b) the associated vbm and Figure 18(c) the green water on deck at the forward perpendicular. The results obtained in the Draupner wave (blue curves) are compared to the results measured in the Peregrine breather (red curves) with Lw/Lpp = 1.3 (cf. Figure 15), respectively. This wave sequence has been chosen due to the fact that the extreme wave at target location features almost the same extraordi‐ narily high wave crest height, but it should be mentioned that the Peregrine breather is also characterized by deeper preceding and following trough as well as shorter up- and downcrossing wave periods, i.e., the Peregrine breather is higher and steeper. Following, pressure sensors at bow and stern are analyzed calculating the exceedance probabilities of the pressure peaks. For this purpose, three analytical models, Weibull, GEV, and Gumbel distributions, were applied. Figure 20 and Figure 21 show the time series of the measured pressures and the identified peaks for the LNGC for Fn=0 in NYW, on the bow and the stern, respectively. Correspondingly, the exceedance probabilities of the peaks are compared using the three analytical models, Weibull, GEV, and Gumbel dis‐ tributions. The total number of the identified peaks of sen‐ sor 2 is small because it is mounted on the upper part of the flare. When the location is lower (for example, pres‐ sure 5), the identified peaks are more. 3.4.1.1 The Akhmediev breather Comparing to the Draupner wave with the Peregrine breather solution (Lw/Lpp= 1.3) where the maxima are 4.45 m, 10.41° and 1.78×109 kN and minima are 5.43 m, 8.56°, and 2.14×109 kN, shows more severity with pitch motion and VBM, although the peaks of the heave motion are 16% (positive) and 18% (negative) lower. 3.4.2 Real world extreme wave reproduction The results show that the breather solutions are good alternatives to the real work abnormal waves, with regards to the slamming load issue. treme values of the pressure peak were all higher in the case when Fn=0.07. The Weibull distribution model pro‐ vides the highest values, and the Gumbel model predicts the smallest for all the pressures. The differences in the es‐ timation between the three models are in general higher as well when the ship has a forward speed. In general, the ex‐ treme pressure is higher at the locations near the calm wa‐ ter surface and is lower at the upper of the flare. Though the total number of the identified pressure peaks on the stern is larger and the geometry of the local structure is flatter, the extreme values there are not higher than the ones on the bow. The maximum peak values of the four pressure sen‐ sors are 0.88 bar, 1.03 bar, 0.67 bar, and 0.89 bar. These values are consistent quite well with the ones from the Breather solutions. For example, the ones from the Pere‐ 3.4.2 Real world extreme wave reproduction As for the ones at the stern, the total number of the identified peaks is gener‐ ally larger due to the low draft at the stern. It is observed that the Weibull distribution fits better for the pressure at the bow and the GEV distribution is more suitable for the ones at the stern. Comparing the measured loads (Figure 18(b)) and green water column on deck (Figure 18(c)) reveals that both extreme waves are dangerous events for the ship. For the Draupner wave impact, the measured vbm reaches the design vertical wave bending moment and the green water column height on deck is with approximately 8 m also impressive. But the impact of the Peregrine breather is more severe result‐ ing in higher hogging and sagging loads as well as green water column height on deck. The main reason for this result is the aforementioned difference between wave height and steepness as well as the wave length of the breather extreme wave as it is more critical regarding the investigated LNGC. This result does not denote that real world registrations reproduced in the wave tank are useless for the evaluation of wave-structure interaction due to the fact that they rep‐ The same analysis was performed by Wang and Guedes Soares (2022a) on the measurements from the case with Fn=0.07, showing the same trends. The extreme values of the pressure peaks obtained using the three distribution models were compared as well. It was found that the ex‐ M. 3.4.2 Real world extreme wave reproduction Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 67 20 15 10 5 0 −5 −10 50 100 150 200 250 300 New Year Wave recorded in the north sea New Year Wave generated in the wave tank 350 400 450 500 550 t (s) ζ (t) (m) Figure 17　Draupner extreme wave-Comparison of measured model wave train at target position and the recorded sequence at the Draupner platform (full scale) 20 15 10 5 0 −5 −10 50 100 150 200 250 300 New Year Wave recorded in the north sea New Year Wave generated in the wave tank 350 400 450 500 550 t (s) ζ (t) (m) Figure 17　Draupner extreme wave-Comparison of measured model wave train at target position and the recorded sequence at the Draupner platform (full scale) Figure 17　Draupner extreme wave-Comparison of measured model wave train at target position and the recorded sequence at the Draupner platform (full scale) 15 10 520 540 560 580 600 ( ) S f l ti t f d di l t (s) 620 640 Peregrine breather Draupner wave MWV 660 680 700 ζ (t) (m) 5 0 −5 −10 20 15 10 520 540 560 580 600 (a) Surface elevation at forward perpendicular t (s) 620 640 Peregrine breather Draupner wave MWV 660 680 520 540 560 580 600 (b) Vertical bending moment at Lpp=2 t (s) 620 640 660 680 700 700 700 520 540 560 580 600 (c) Green water on deck at forward perpendicular t (s) 620 640 660 680 ζ (t) (m) vbm (t) (kN·m) 5 0 −5 −0.5 −1.0 12 10 8 6 4 2 0 −1.5 −2.0 −10 20 1.5 2.0 1.0 0.5 0 ζgreen water (t) (m) Figure 18　Results of the model tests in the Draupner wave vs. Peregrine breather extreme wave (c) Green water on deck at forward perpendicular Figure 18　Results of the model tests in the Draupner wave vs. Peregrine breather extreme wave grine breather solution with Lw/Lpp= 1.1 have around 0.61%, −2.39%, 0.9% and 2.28% difference relative to the ones from NYW. The results show that the breather solutions are good alternatives to the real work abnormal waves, with regards to the slamming load issue. grine breather solution with Lw/Lpp= 1.1 have around 0.61%, −2.39%, 0.9% and 2.28% difference relative to the ones from NYW. 4 Conclusion This paper presented a comprehensive experimental study on the impact of extreme waves on a LNG carrier. It is shown that investigations in the frequency domain, i.e., in transient wave packets and regular waves, are essential to evaluate the overall characteristics in terms of the RAO. In particular the (non-linear) behavior in higher, steeper waves Journal of Marine Science and Application 68 Journal of Marine Science and Application can be investigated systematically in regular waves. Further‐ more, the study revealed that model tests in design extreme waves (i.e., predefined wave length) using breather solutions are a great alternative to search for critical wave sequences in terms of wave length and height. From this it follows that the detected absolute vbm values in breather-type extreme waves of the same wave length. In addition, the impact of the extreme waves resulted in impressive amount of green water on deck. Investigations in a real world freak wave reproduction- the Draupner wave-showed the same trend: the impact of the extreme wave is severe and dangerous resulting in large Figure 20　The identified peaks of the pressures at the bow at Fn=0 and the corresponding exceedance probability Figure 19　Exceedance probability of peaks for the motions and VBM for the LNGC in the Draupner wave at Fn=0 68 Figure 19　Exceedance probability of peaks for the motions and VBM for the LNGC in the Draupner wave at Fn=0 Figure 19　Exceedance probability of peaks for the motions and VBM for the LNGC in the Draupner wave at Fn=0 Figure 20　The identified peaks of the pressures at the bow at Fn=0 and the corresponding exceedance probability Figure 20　The identified peaks of the pressures at the bow at Fn=0 and the corresponding exceedance probability peaks of the pressures at the bow at Fn=0 and the corresponding exceedance probability waves of the same wave length. In addition, the impact of the extreme waves resulted in impressive amount of green water on deck. can be investigated systematically in regular waves. Further‐ more, the study revealed that model tests in design extreme waves (i.e., predefined wave length) using breather solutions are a great alternative to search for critical wave sequences in terms of wave length and height. Appendix B Details on pressure transducers extreme waves show that larger loads can be observed in the breather-type extreme waves due to the fact that the wave length can be adjusted in order to meet a critical wave length (in contrast to a real world registration). Altogether, this study shows that the breather solutions are a powerful tool for the generation of tailored freak waves of certain critical wave lengths for wave/structure investigations on different subjects, e.g., local and global loads, green water effects as well as air gap investigations and an enrichment for the test portfolio regarding tailored critical wave sequences. Table B2 presents the positions of the pressure transduc‐ ers on the hull of the LNGC. The information only pro‐ vides the X and Z-coordinates-the corresponding Y-posi‐ tion results from the according hull geometry. The posi‐ tions are given in meters in model scale with the origin lo‐ cated at the aft perpendicular at keel level. For the model tests, the pressure transducer HKM375- 1.7 Bar A from Kulite were installed. The signal amplifiers for the pressure transducers were custom made, designed and manufactured in-house. The motivation for develop‐ ing in-house signal amplifier lied in the application area. The dynamic pressures were expected to be in a very small measuring range so that only very low-noise amplifier can accurately detect the measured signals. Figure B1 presents time traces of two pressure transducers exemplary. It is shown that the noise level is significantly lower compared to the measured values. Acknowledgement The work contributes to the Strategic Research Plan of the Centre for Marine Technology and Ocean Engineering (CENTEC). CENTEC is financed by the Portuguese Foundation for Science and Technology (Fundaço para a Ciência e Tecnologia-FCT) under contract UIDB/UIDP/00134/2020. Funding This paper is published based on the experimental work performed during the project EXTREME SEAS, which was funded by the European Commission, under the Grant agreement No. 234175. Nevertheless, experimental uncertainties were observed in the model tests, i. e, some of the pressure transducers showed an unusual behavior during the model tests (cf. Figure B1(a)). This can be related to the different tempera‐ ture compensation behavior of the sensors, even though all sensors should provide the same specifications - a circum‐ stance which has to be taken into account carefully for the interpretation of the measured results. The installed pres‐ sure transducers can be classified into three different types 4 Conclusion From this it follows that the detected absolute vbm values in breather-type extreme waves are at least 15% higher for hogging condition and 30% higher for sagging condition in comparison to regular Investigations in a real world freak wave reproduction- the Draupner wave-showed the same trend: the impact of the extreme wave is severe and dangerous resulting in large loads and huge amount of green water on deck. Evaluating the results in the Draupner wave and in the breather-type M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 69 Figure 21　The identified peaks of the pressures at the stern at Fn=0 and the corresponding exceedance probability Figure 21　The identified peaks of the pressures at the stern at Fn=0 and the corresponding exceedance probability Appendix A Details on ship model mass distribution 2 5 8 11 14 17 20 23 26 29 32 35 38 x (m) 2.700 2.670 2.640 2.610 2.610 2.580 2.550 2.520 2.520 2.490 0.000 0.060 2.580 y (m) 0.220 0.195 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.140 0.140 0.267 No. 3 6 9 12 15 18 21 24 27 30 33 36 x (m) 2.700 2.670 2.640 2.610 2.580 2.580 2.550 2.520 2.490 2.490 0.000 2.580 y (m) 0.195 0.170 0.170 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.125 0.000 Table A1　Detailed mass distribution, moments of inertia and center of gravity (model scale) Table B2　Positions of the pressure transducers on the LNGC No. 1 4 7 10 13 16 19 22 25 28 31 34 37 x (m) 2.730 2.670 2.640 2.640 2.610 2.580 2.550 2.550 2.520 2.490 0.000 0.030 2.700 y (m) 0.220 0.220 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.195 0.164 0.140 0.267 No. 2 5 8 11 14 17 20 23 26 29 32 35 38 x (m) 2.700 2.670 2.640 2.610 2.610 2.580 2.550 2.520 2.520 2.490 0.000 0.060 2.580 y (m) 0.220 0.195 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.140 0.140 0.267 No. 3 6 9 12 15 18 21 24 27 30 33 36 x (m) 2.700 2.670 2.640 2.610 2.580 2.580 2.550 2.520 2.490 2.490 0.000 2.580 y (m) 0.195 0.170 0.170 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.125 0.000 Table B2　Positions of the pressure transducers on the LNGC Figure B1　Time trace of two pressure transducers with normal (B1(b)) and disputable (B1(a)) behavior Besides systematic uncertainties of some pressure sen‐ sors, also the random experimental uncertainties of slam‐ ming pressures were estimated (Wang and Guedes Soares 2022b). For this purpose, an irregular sea state with a signifi‐ cant wave height of 11.5 m and a peak period of 12 s de‐ scribed by the JONSWAP spectrum (case IRREGULAR20 in Table C3) was considered. Altogether, five different test runs (different phase seeds) were conducted. Figure B2 presents the time series and the identified peaks for the measurements from two pressure sensors (Figure B2(a) for No. 03 and Figure B2(c) for No. 06-cf. Figure 2(a) and Table B2) in sea state IRREGULAR20 for phase seed #1. Figure B2(b) indicate the goodness of fit for GEV distribu‐ tion on the selected peaks for pressure sensor No. 03, and Figure B2(d) for pressure sensor No. Appendix A Details on ship model mass distribution 2 5 8 11 14 17 20 23 26 29 32 35 38 x (m) 2.700 2.670 2.640 2.610 2.610 2.580 2.550 2.520 2.520 2.490 0.000 0.060 2.580 y (m) 0.220 0.195 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.140 0.140 0.267 No. 3 6 9 12 15 18 21 24 27 30 33 36 x (m) 2.700 2.670 2.640 2.610 2.580 2.580 2.550 2.520 2.490 2.490 0.000 2.580 y (m) 0.195 0.170 0.170 0.195 0.220 0.145 0.170 0.195 0.220 0.145 0.125 0.000 of behavior: • Regular behavior: no shifting pressure offset (cf. Fig‐ ure B1(b)) • Disputable behavior: a fast shifting offset regarding the reference pressure during each wave impact (cf. Figure B1, top), and a slow decay to the original reference pres‐ sure after each wave impact • Acceptable behavior: a fast shifting offset regarding the reference pressure at the FIRST wave impact, constant offset through the entire test run to be regarded as the new reference pressure level, i.e., no decay to the original refer‐ ence pressure Besides systematic uncertainties of some pressure sen‐ sors, also the random experimental uncertainties of slam‐ ming pressures were estimated (Wang and Guedes Soares 2022b). For this purpose, an irregular sea state with a signifi‐ cant wave height of 11.5 m and a peak period of 12 s de‐ scribed by the JONSWAP spectrum (case IRREGULAR20 in Table C3) was considered. Altogether, five different test runs (different phase seeds) were conducted. Figure B2 presents the time series and the identified peaks for the measurements from two pressure sensors (Figure B2(a) for No. 03 and Figure B2(c) for No. 06-cf. Figure 2(a) and Table B2) in sea state IRREGULAR20 for phase seed #1. Figure B2(b) indicate the goodness of fit for GEV distribu‐ tion on the selected peaks for pressure sensor No. 03, and Figure B2(d) for pressure sensor No 06 It was found that 20 100 110 120 130 140 t (s) 150 (a) Pressure sensor No. 02 (b) Pressure sensor No. Appendix A Details on ship model mass distribution 03 160 170 180 100 110 120 130 140 t (s) 150 160 170 180 15 10 Pressure (bar) 5 0 −5 20 15 10 Pressure (bar) 5 0 −5 Figure B1　Time trace of two pressure transducers with normal (B1(b)) and disputable (B1(a)) behavior Table A1　Detailed mass distribution, moments of inertia and center of gravity (model scale) Items Fore ship Aft ship Force transducer Force transducer Force transducer Trim weight 1 Trim weight 2 Trim weight 3 Trim weight 4 Trim weight 5 Trim weight 6 Trim weight 7 Trim weight 8 Trim weight 9 Trim weight 10 Trim weight 11 Trim weight 12 Trim weight 13 Equipped ship model Mass (kg) 29.600 24.500 1.800 1.800 1.800 1.400 4.980 10.040 3.835 3.835 3.835 3.835 10.040 2.020 0.340 0.057 0.057 0.057 103.831 Center of gravity x (m) 1.827 0.784 1.335 1.335 1.335 2.118 2.118 1.769 1.256 1.255 1.104 1.103 0.877 0.722 1.318 1.973 0.466 0.459 1.355 y (m) 0.000 0.000 0.183 −0.183 0.000 0.000 0.000 0.000 0.074 0.074 −0.074 −0.074 0.000 0.000 0.000 0.000 0.218 −0.218 0.000 z (m) 0.151 0.174 0.283 0.283 −0.015 0.042 0.025 0.059 0.059 0.059 0.059 0.059 0.059 0.019 0.308 0.311 0.501 0.501 0.118 Moment of inertia Jxx (kg·m2) 0.957 0.917 0.109 2 0.109 2 0.031 7 0.001 9 0.012 1 0.032 2 0.005 9 0.005 9 0.005 9 0.005 9 0.023 0 0.003 0 0.038 1 0.000 0 0.000 0 0.000 0 2.682 Jyy (kg·m2) 5.283 5.064 0.050 0 0.050 0 0.032 4 0.001 9 0.012 1 0.022 9 0.010 0 0.010 0 0.010 0 0.010 0 0.032 1 0.003 0 0.000 1 0.000 0 0.000 0 0.000 0 34.811 Jzz (kg·m2) 5.654 4.240 0.060 7 0.060 7 0.000 7 0.003 9 0.023 7 0.031 0 0.010 3 0.010 3 0.010 3 0.010 3 0.031 0 0.006 1 0.038 1 0.000 0 0.000 0 0.000 0 34.165 Table B2　Positions of the pressure transducers on the LNGC No. 1 4 7 10 13 16 19 22 25 28 31 34 37 x (m) 2.730 2.670 2.640 2.640 2.610 2.580 2.550 2.550 2.520 2.490 0.000 0.030 2.700 y (m) 0.220 0.220 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.195 0.164 0.140 0.267 No. Appendix A Details on ship model mass distribution Table A1 presents the detailed mass distribution, the cor‐ responding moments of inertia and the center of gravity. Journal of Marine Science and Application 70 of behavior: • Regular behavior: no shifting pressure offset (cf. Fig‐ ure B1(b)) • Disputable behavior: a fast shifting offset regarding the reference pressure during each wave impact (cf. Figure B1, top), and a slow decay to the original reference pres‐ sure after each wave impact • Acceptable behavior: a fast shifting offset regarding the reference pressure at the FIRST wave impact, constant offset through the entire test run to be regarded as the new reference pressure level, i.e., no decay to the original refer‐ ence pressure Besides systematic uncertainties of some pressure sen‐ sors, also the random experimental uncertainties of slam‐ ming pressures were estimated (Wang and Guedes Soares 2022b). For this purpose, an irregular sea state with a signifi‐ cant wave height of 11.5 m and a peak period of 12 s de‐ scribed by the JONSWAP spectrum (case IRREGULAR20 in Table C3) was considered. Altogether, five different test runs (different phase seeds) were conducted. Figure B2 presents the time series and the identified peaks for the measurements from two pressure sensors (Figure B2(a) for No. 03 and Figure B2(c) for No. 06-cf. Figure 2(a) and Table B2) in sea state IRREGULAR20 for phase seed #1. Figure B2(b) indicate the goodness of fit for GEV distribu‐ tion on the selected peaks for pressure sensor No. 03, and Figure B2(d) for pressure sensor No. 06. It was found that 20 100 110 120 130 140 t (s) 150 (a) Pressure sensor No. 02 (b) Pressure sensor No. Appendix A Details on ship model mass distribution 03 160 170 180 100 110 120 130 140 t (s) 150 160 170 180 15 10 Pressure (bar) 5 0 −5 20 15 10 Pressure (bar) 5 0 −5 Figure B1　Time trace of two pressure transducers with normal (B1(b)) and disputable (B1(a)) behavior Table A1　Detailed mass distribution, moments of inertia and center of gravity (model scale) Items Fore ship Aft ship Force transducer Force transducer Force transducer Trim weight 1 Trim weight 2 Trim weight 3 Trim weight 4 Trim weight 5 Trim weight 6 Trim weight 7 Trim weight 8 Trim weight 9 Trim weight 10 Trim weight 11 Trim weight 12 Trim weight 13 Equipped ship model Mass (kg) 29.600 24.500 1.800 1.800 1.800 1.400 4.980 10.040 3.835 3.835 3.835 3.835 10.040 2.020 0.340 0.057 0.057 0.057 103.831 Center of gravity x (m) 1.827 0.784 1.335 1.335 1.335 2.118 2.118 1.769 1.256 1.255 1.104 1.103 0.877 0.722 1.318 1.973 0.466 0.459 1.355 y (m) 0.000 0.000 0.183 −0.183 0.000 0.000 0.000 0.000 0.074 0.074 −0.074 −0.074 0.000 0.000 0.000 0.000 0.218 −0.218 0.000 z (m) 0.151 0.174 0.283 0.283 −0.015 0.042 0.025 0.059 0.059 0.059 0.059 0.059 0.059 0.019 0.308 0.311 0.501 0.501 0.118 Moment of inertia Jxx (kg·m2) 0.957 0.917 0.109 2 0.109 2 0.031 7 0.001 9 0.012 1 0.032 2 0.005 9 0.005 9 0.005 9 0.005 9 0.023 0 0.003 0 0.038 1 0.000 0 0.000 0 0.000 0 2.682 Jyy (kg·m2) 5.283 5.064 0.050 0 0.050 0 0.032 4 0.001 9 0.012 1 0.022 9 0.010 0 0.010 0 0.010 0 0.010 0 0.032 1 0.003 0 0.000 1 0.000 0 0.000 0 0.000 0 34.811 Jzz (kg·m2) 5.654 4.240 0.060 7 0.060 7 0.000 7 0.003 9 0.023 7 0.031 0 0.010 3 0.010 3 0.010 3 0.010 3 0.031 0 0.006 1 0.038 1 0.000 0 0.000 0 0.000 0 34.165 Table B2　Positions of the pressure transducers on the LNGC No. 1 4 7 10 13 16 19 22 25 28 31 34 37 x (m) 2.730 2.670 2.640 2.640 2.610 2.580 2.550 2.550 2.520 2.490 0.000 0.030 2.700 y (m) 0.220 0.220 0.220 0.145 0.170 0.195 0.220 0.145 0.170 0.195 0.164 0.140 0.267 No. Appendix A Details on ship model mass distribution 06. It was found that of behavior: • Regular behavior: no shifting pressure offset (cf. Fig‐ ure B1(b)) • Disputable behavior: a fast shifting offset regarding the reference pressure during each wave impact (cf. Figure B1, top), and a slow decay to the original reference pres‐ sure after each wave impact • Acceptable behavior: a fast shifting offset regarding the reference pressure at the FIRST wave impact, constant offset through the entire test run to be regarded as the new reference pressure level, i.e., no decay to the original refer‐ ence pressure M. Klein et al.: Experimental Study on the Effect of Extreme Waves on a LNG Carrier 71 Figure B2　Time series of the pressures and the diagnostic plots of goodness of fit for generalized extreme value distribution of the identified peak values for pressure sensors No. 03 and No. 06 Figure B2　Time series of the pressures and the diagnostic plots of goodness of fit for generalized extreme value distribution of the identified peak values for pressure sensors No. 03 and No. 06 the time series of from phase seed #1 and phase seed #2 were somehow different. The peaks of the pressures from sensors No. 03 and No. 06 are both lower in the measure‐ ments from phase seed #2. The extreme value, the average value of the largest 1/10, the average of the largest 1/3 and the mean value of the peaks were calculated, and the ran‐ dom standard uncertainties were calculated as the standard deviation of the metric across the 5 different phase seeds, divided by the square root of the number of observations. The percentages of the relative random uncertainty on the pressure peaks obtained using the data from the five different phase seeds and the ones excluding phase seed #2 are shown in Figure B3. The uncertainties are much lower when the data from phase seed #2 is excluded. The study also showed that the GEV distribution model is suitable to study the peak values of the pressures of the hull subjected to extreme waves. The random uncertainties for pressure peaks are slightly higher, eg. 5.94% on the extreme value of pressure sensor 03 for the ship with zero speed (Fn=0). However, the values increase greatly for the ship with Fn=0.07, e.g., is 11.8% for the same metrics. The complete analysis can be found in Wang and Guedes Soares (2022b). Figure B3　Experimental random uncertainties on the pressure peaks Figure B3　Experimental random uncertainties on the pressure peaks Figure B3　Experimental random uncertainties on the pressure peaks 72 Journal of Marine Science and Application Appendix C Details on investigated irregular sea states Table C3　Positions of the pressure transducers on the LNGC No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Synonym IRREGULAR13 IRREGULAR14 IRREGULAR15 IRREGULAR16 IRREGULAR17 IRREGULAR18 IRREGULAR19 IRREGULAR20 IRREGULAR21 IRREGULAR02 IRREGULAR06 IRREGULAR10 IRREGULAR12 IRREGULAR22 IRREGULAR23 IRREGULAR24 IRREGULAR25 IRREGULARTUB Hs (m) 16.5 16.5 3 9.7 9.7 9.7 11.5 11.5 11.5 8.5 10.5 13.5 16.5 8.5 12.5 13.5 14.5 11 Tp (s) 16.1 15.9 12 12 12 12 12 12 12 10.92 12.21 13.49 14.78 9.64 12.21 12.21 13.49 11.4 γ 1 3.3 3.3 1 3.3 6 1 3.3 6 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 Phase seeds 5 3 1 3 5 5 3 5 6 1 1 1 1 1 1 1 1 1 Appendix C Details on investigated irregular sea states Cherneva Z, Guedes Soares C (2008) Non-linearity and non-stationarity of the new year abnormal wave. 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Conference on the Behaviour of Offshore Structures (BOSS), Delft, The Nether‐ lands, 269-285 Clauss G, Lehmann E, Östergaard C (1992) Offshore Structures, Volume 1: Conceptual Design and Hydrodynamics. Springer Verlag London Clauss G, Steinhagen U (1999) Numerical simulation of nonlinear transient waves and its validation by laboratory data. Proceedings of 9th International Offshore and Polar Engineering Conference (ISOPE), Vol. III, Brest, France, 368-375 Clauss GF, Klein M, Dudek M (2010) Influence of the bow shape on loads in high and steep waves. 29th International Conference on Ocean, Offshore and Arctic Engineering: Volume 2, Shanghai, Chi‐ na, 159-170 Clauss GF, Klein M, Dudek M, Onorato M (2012) Application of breather solutions for the investigation of wave/structure interac‐ tion in high steep waves. International Conference on Offshore Mechanics and Arctic Engineering, Volume 44892, Rio de Janei‐ ro, Brazil, 123-135 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and thesource, provide a link to the Creative Commons licence, and indicateif changes were made. The images or other third party material in thisarticle are included in the article’s Creative Commons licence, unlessindicated otherwise in a credit line to the material. If material is notincluded in the article’s Creative Commons licence and your intendeduse is not permitted by statutory regulation or exceeds the permitteduse, you will need to obtain permission directly from the copyrightholder. To view a copy of this licence, visit http:// creativecommons.org/licenses/by/4.0/. 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https://openalex.org/W2952318715	https://digitalcommons.uri.edu/cgi/viewcontent.cgi?article=1034&context=che_facpubs	English	null	A Carbon Nanotube Optical Reporter Maps Endolysosomal Lipid Flux	ACS nano	2,017	cc-by	16,493	University of Rhode Island University of Rhode Island DigitalCommons@URI DigitalCommons@URI Chemical Engineering Follow this and additional works at: https://digitalcommons.uri.edu/che_facpubs Part of the Chemical Engineering Commons Disciplines Disciplines Chemical Engineering Disciplines Disciplines Chemical Engineering Citation/Publisher Attribution Citation/Publisher Attribution Citation/Publisher Attribution Citation/Publisher Attribution Jena, P. V., Roxbury, D., Galassi, T. V., Akkari, L., Horoszko, C. P., Iaea, D. B., Budhathoki-Uprety, J., Pipalia, N., Haka, A. S., Harvey, J. D., Mittal, J., Maxfield, F. R., Joyce, J. A., & Heller, D. A. (2017). A Carbon Nanotube Optical Reporter Maps Endolysosomal Lipid Flux. ACS Nano, 11(11), 10689-10703. doi: 10.1021/acsnano.7b04743 Available at: http://dx.doi.org/10.1021/acsnano.7b04743 Available at: http://dx.doi.org/10.1021/acsnano.7b04743 This Article is brought to you by the University of Rhode Island. It has been accepted for inclusion in Chemical Engineering Faculty Publications by an authorized administrator of DigitalCommons@URI. For more information, please contact digitalcommons-group@uri.edu. For permission to reuse copyrighted content, contact the author directly. Authors Authors Jena V. Prakrit, Daniel E. Roxbury, Thomas V. Galassi, Leila Akkari, Christopher P. Horoszko, David B. Iaea, Januka Budhathoki-Uprety, Nina Pipalia, Abigail S. Haka, Jackson D. Harvey, Jeetain Mittal, Frederick R. Maxfield, Johanna A. Joyce, and Daniel A. Heller The University of Rhode Island Faculty have made this article openly available. The University of Rhode Island Faculty have made this article openly available. Please let us know Please let us know how Open Access to this research benefits you. how Open Access to this research benefits you. This is a pre-publication author manuscript of the final, published article. This article is made available under the terms and conditions applicable towards Open Access Policy Articles, as set forth in our Terms of Use. le is available at DigitalCommons@URI: https://digitalcommons.uri.edu/che_facpubs/32 A Carbon Nanotube Optical Reporter Maps Endolysosomal Lipid Flux Prakrit V. Jena†1, Daniel Roxbury†2, Thomas V. Galassi†1,3, Leila Akkari1,4, Christopher P. Horoszko1,3, David B. Iaea3, Januka Budhathoki-Uprety1, Nina Pipalia3, Abigail S. Haka3, Jackson Dean Harvey1,3, Jeetain Mittal5, Frederick R. Maxfield3, Johanna A. Joyce1,3,6, Daniel A. Heller1,3* 1Memorial Sloan Kettering Cancer Center, New York, NY 10065 1Memorial Sloan Kettering Cancer Center, New York, NY 10065 2Department of Chemical Engineering, University of Rhode Island, Kingston, RI 02881 3Weill Cornell Medical College, New York, NY 10065 4Division of Tumor Biology & Immunology, the Netherlands Cancer Institute, Amsterdam, the Netherlands 5Department of Chemical and Biomolecular Engineering, Lehigh University, Bethlehem, PA 18015 6Ludwig Center for Cancer Research, University of Lausanne, Switzerland, CH 1066 † Contributed equally to this work Corresponding author. Email: hellerd@mskcc.org KEYWORDS: singe-walled carbon nanotubes, lipids, sensors, near-infrared imaging, single-cell ABSTRACT: Lipid accumulation within the lumen of endolysosomal vesicles is observed in various pathologies including atherosclerosis, liver disease, neurological disorders, lysosomal storage disorders, and cancer. Current methods cannot measure lipid flux specifically within the lysosomal lumen of live cells. We developed an optical reporter, composed of a ABSTRACT: Lipid accumulation within the lumen of endolysosomal vesicles is observed in various pathologies including atherosclerosis, liver disease, neurological disorders, lysosomal storage disorders, and cancer. Current methods cannot measure lipid flux specifically within the lysosomal lumen of live cells. We developed an optical reporter, composed of a photoluminescent carbon nanotube of a single chirality, which responds to lipid accumulation via modulation of the nanotube’s optical bandgap. The engineered nanomaterial, composed of short- single stranded DNA and a single nanotube chirality, localizes exclusively to the lumen of endolysosomal organelles without adversely affecting cell viability or proliferation, or organelle morphology, integrity, or function. The emission wavelength of the reporter can be spatially resolved from within the endolysosomal lumen to generate quantitative maps of lipid content in live cells. Endolysosomal lipid accumulation in cell lines, an example of drug-induced phospholipidosis (DIPL), was observed for multiple drugs in macrophages, and measurements of patient-derived Niemann-Pick type C fibroblasts identified lipid accumulation and phenotypic reversal of this lysosomal storage disease. Single-cell measurements using the reporter discerned sub-cellular differences in equilibrium lipid content, illuminating significant intracellular heterogeneity among endolysosomal organelles of differentiating bone marrow-derived monocytes. Single-cell kinetics of lipoprotein-derived cholesterol accumulation within macrophages revealed rates that differed among cells by an order of magnitude. This carbon nanotube optical reporter of endolysosomal lipid content in live cells confers additional capabilities for drug development processes and the investigation of lipid-linked diseases. 1Memorial Sloan Kettering Cancer Center, New York, NY 10065 Endosomes and lysosomes are vacuolar organelles responsible for the breakdown of lipids, proteins, sugars, and other cellular materials.1 The failure to catabolize or export lysosomal contents can result in lysosomal storage disorders (LSDs), a family of approximately 50 diseases characterized by the accumulation of undigested substrates, such as lipids and glycoproteins, within the endolysosomal lumen due to an inherited defect in a single protein.2,3 Lipid accumulation in the endolysosomal lumen is observed in many LSDs as well as during atherosclerotic foam cell formation,4 the transition from steatosis to non-alcoholic steatohepatitis,5 in multiple neurological disorders,6 cancer,7 and drug-induced phospholipidosis (DIPL).8 The search for small molecule therapeutics against LSDs, as well as our understanding of the aforementioned diseases is hampered by the limited number of tools available to assay lipid content exclusively within the endolysosomal organelles of live cells.9 lysosomal contents can result in lysosomal storage disorders (LSDs), a family of approximately 50 diseases characterized by the accumulation of undigested substrates, such as lipids and glycoproteins, within the endolysosomal lumen due to an inherited defect in a single protein.2,3 Lipid accumulation in the endolysosomal lumen is observed in many LSDs as well as during atherosclerotic foam cell formation,4 the transition from steatosis to non-alcoholic steatohepatitis,5 in multiple neurological disorders,6 cancer,7 and drug-induced phospholipidosis Although multiple classes of sensors and imaging modalities exist to study lipids, current probes are limited in their capabilities. Stains such as LipidTox can detect the general accumulation of lipids within cells10 but are not organelle-specific. Fluorophore-conjugated and intrinsically fluorescent lipid analogs are used for analyzing lipid trafficking.10 Lipid analogs, synthesized by conjugation of a fluorophore to a modified lipid, allow the tracking of uptake and incorporation of lipids into the cell membrane.11,12 Lipid dynamics can be tracked in live cells using fluorescent proteins fused with lipid-binding domains, however the expressed domains can hamper the native function of the lipid.13 Environmentally-sensitive fluorophores were recently developed, which can respond to lipid order14 in cell membranes undergoing processes such as endosomal maturation,15 while another family of polarity-sensitive probes, can integrate into lysosomal membranes and detect changes in the overall polarity,16 The bulk of these technologies are useful for studying lipids present within a biological membrane, but, to the best of our knowledge, no existing probe specifically localizes to the lumen of endolysosomal organelles and reports on the lipid content of its immediate environment. 1Memorial Sloan Kettering Cancer Center, New York, NY 10065 To develop a biocompatible fluorescent reporter for lipids in the endolysosomal lumen, we investigated the physicochemical properties of single-walled carbon nanotubes (SWCNTs). Semiconducting SWCNTs emit highly photostable and narrow-bandwidth near-infrared photoluminescence,17 which is sensitive to local perturbations.18 The availability of multiple species with different emission properties can facilitate multiplexed imaging.19 The SWCNT emission energy responds to the solvent environment20 via solvatochromic energy shifts.21 This response has been used to detect conformational polymorphism22 of DNA and the nuclear environment in live cells,23 as well as microRNA,24 via shifts down to ≤1 nm. While the self- assembly of lipid derivatives on carbon nanotubes was observed over 14 years ago,25 the optical response of fluorescent carbon nanotubes to fatty acids has been noted more recently.26 Due to their applications in biological sensing and imaging applications,27 the biocompatibility of carbon nanotubes has been a subject of much investigation.28,29 A recent comprehensive review concluded that the biocompatibility of single-walled carbon nanotubes is dependent on how the nanomaterial sample is processed and functionalized.30 In particular, multi-walled carbon nanotubes and long single-walled carbon nanotubes or nanotube preparations containing impurities have documented toxic effects on live cells.31 Here, we present a biocompatible carbon nanotube optical reporter of lipids within the endolysosomal lumen of live cells. Composed of a non-covalent complex consisting of an amphiphilic polymer and a single (n,m) species (chirality) of carbon nanotube, the reporter exhibits a solvatochromic shift of over 13 nm in response to biological lipids. In mammalian cells, the reporter remains within the endolysosomal pathway and localizes specifically to the lumen of endolysosomal organelles without adversely affecting organelle morphology, integrity, capacity to digest substrates, or cell viability or proliferation. Using near-infrared hyperspectral microscopy, we spatially resolved the solvatochromic response of the reporter to lipids in the endolysosomal lumen and obtained quantitative lipid maps of live cells with sub-cellular resolution. Emission from the reporter identified the lysosomal storage disease Niemann-Pick type C (NPC) in fibroblasts from an NPC patient. Furthermore, the reporter benchmarked treatment, exhibiting a distinct signal reversal upon administration of hydroxypropyl-β- cyclodextrin, a drug that reverses the disease phenotype. Additionally, endolysosomal lipid accumulation was detected using spectroscopy alone, in a 96-well plate format compatible with high-throughput drug screening. Using the reporter, single-cell kinetic measurements in a macrophage model system for lysosomal lipid accumulation identified a sub-population of cells that was both significantly lipid deficient and slower to accumulate cholesterol in the lysosomal lumen. 1Memorial Sloan Kettering Cancer Center, New York, NY 10065 In the context of primary monocyte differentiation into macrophages, we discovered that, as the lipid content in the lumen increases, individual endolysosomal organelles within single cells accumulate lipids at different rates. Thus, our optical reporter enables quantitative imaging and high-throughput measurement of the lipid content in the endolysosomal lumen of live cells. Carbon nanotube optical response to lipids To develop a reporter of endolysosomal lipid content, we first identified a structurally- defined DNA-carbon nanotube complex that responds optically to lipids. Carbon nanotubes were non-covalently functionalized with specific ssDNA oligonucleotides to facilitate separation using ion-exchange chromatography,32 resulting in suspensions of single-chirality DNA-nanotube complexes. The introduction of low density lipoprotein (LDL), a biochemical assembly composed of lipids and proteins,33 induced a decrease in emission wavelength (blue shift) that ranged from 0 to 13 nm (Figure S1). The largest solvatochromic response was observed from the (8,6) nanotube spectral band complexed with ss(GT)6, a short oligonucleotide which facilitates separation of the (8,6) species (Figure S2).32 The isolated ss(GT)6-(8,6) complex exhibited absorption bands at 730 nm and 1200 nm and a single photoluminescence emission peak at 1200 nm (Figure 1a, S2-3). Previous work by our lab indicates that the mean length of ss(GT)6-(8,6) complexes following the preparation method used is 88.9 nm.34 We also recently experimentally identified ss(GT)6-(8,6) as a sequence-chirality pair that optimally maximized both the high dynamic range of optical modulation by a lipid-like surfactant (SDC) and the stability of the DNA-nanotube complex.35 The ss(GT)6-(8,6) complex was characterized by measuring the optical response to several classes of biomolecules and water-soluble lipid analogs. Cholesterol-conjugated polyethylene glycol (PEG), a water-soluble analog of cholesterol, induced a ~ 10 nm decrease in the emission wavelength of the complex at both 2 and 24 hours, while saturating concentrations of bovine serum albumin (BSA), dsDNA from salmon testes, or carboxymethyl cellulose had no measurable effect (Figure S4). The nanotube emission responded rapidly to cholesterol-PEG at 2 hours, but at equilibrium, two different classes of water-soluble lipid analogs elicited equivalent, large blue shifting responses (Figure S5). The optical response of the ss(GT)6-(8,6) complex was monotonic and linear over two orders of magnitude of cholesterol concentrations (Figure 1b). A similar response was observed for both ceramide (Figure S6) and low-density lipoprotein (LDL) (Figure S6), indicating the general sensitivity of ss(GT)6-(8,6) to both water-soluble lipid analogs and native lipids. Carbon nanotube optical response to lipids To probe the underlying mechanism governing the response of the ss(GT)6-(8,6) To probe the underlying mechanism governing the response of the ss(GT)6-(8,6) To probe the underlying mechanism governing the response of the ss(GT)6-(8,6) complex, we examined the dependence of emission on the dielectric environment.20 Using near- To probe the underlying mechanism governing the response of the ss(GT)6-(8,6) l i d h d d f i i h di l i i 20 i complex, we examined the dependence of emission on the dielectric environment.20 Using near- infrared hyperspectral microscopy,19 we obtained the emission spectrum from individual surface- adsorbed ss(GT)6-(8,6) complexes in 7 different solvent environments (Table S1). The peak emission wavelength of the complexes ranged over 20 nm and exhibited a direct correlation with solvent dielectric constant (Figure 1c) with a Spearman correlation of 0.89, p<0.01. This result suggests that the ss(GT)6-(8,6) complex exhibits a distinct solvatochromic response. Figure 1. Optical response of carbon nanotube complexes to lipid environments. (a) Normalized absorption and emission spectra of ss(GT)6-carbon nanotube complexes purified to isolate the (8,6) species. (b) Emission peak wavelength of ss(GT)6-(8,6) nanotube complexes in solution as a function of cholesterol-PEG concentration. Error bars are standard error of the mean, obtained from 3 technical replicates performed for each concentration. (c) Mean emission wavelength of ss(GT)6-(8,6) nanotube complexes exposed to different solvents. Error bars are standard errors of the mean, obtained from 5 technical replicates for each solvent. (d) Frames from all-atom molecular dynamics simulations of equilibrated structures of the ss(GT)6-(8,6) nanotube complex in water, and the same complex equilibrated in the presence of cholesterol or sphingomyelin. (e) Water molecule density as a function of distance from the center of the equilibrated ss(GT)6-(8,6) nanotube complex and the same complex equilibrated in the presence of cholesterol or sphingomyelin. Figure 1. Optical response of carbon nanotube complexes to lipid environments. (a) Normalized absorption and emission spectra of ss(GT)6-carbon nanotube complexes purified to isolate the (8,6) species. (b) Emission peak wavelength of ss(GT)6-(8,6) nanotube complexes in solution as a function of cholesterol-PEG concentration. Error bars are standard error of the mean, obtained from 3 technical replicates performed for each concentration. (c) Mean emission wavelength of ss(GT)6-(8,6) nanotube complexes exposed to different solvents. Error bars are standard errors of the mean, obtained from 5 technical replicates for each solvent. Carbon nanotube optical response to lipids The sample length distribution lies between ultra-short (50 nm) and short (150 nm) nanotubes, which maximizes cellular uptake of fluorescent biocompatible single stranded DNA. This minimizes the key parameters of SWCNT cellular toxicity,30 a topic that is assessed below. The sample length distribution lies between ultra-short (50 nm) and short (150 nm) nanotubes, which maximizes cellular uptake of fluorescent nanotubes while minimizing bundling within cells.38 The observed brightness of structurally- sorted ss(GT)6-(8,6) is intrinsically higher than unsorted DNA-nanotube sensors, as on-resonance excitation at 730 nm efficiently excites every nanotube present. Additionally, this particular sequence-chirality pair is relatively stable, and retains its structural integrity under surfactant exchange.35 The structure, stability, and brightness of ss(GT)6-(8,6), combined with its sensitivity and specificity to lipids over other classes of biomolecules, suggests that it may be applied to live-cell measurements of lipid accumulation. Carbon nanotube optical response to lipids (d) Frames from all-atom molecular dynamics simulations of equilibrated structures of the ss(GT)6-(8,6) nanotube complex in water, and the same complex equilibrated in the presence of cholesterol or sphingomyelin. (e) Water molecule density as a function of distance from the center of the equilibrated ss(GT)6-(8,6) nanotube complex and the same complex equilibrated in the presence of cholesterol or sphingomyelin. For further understanding how lipids interact with the surface of ss(GT)6-(8,6) nanotube complexes to induce a solvatochromic shift, we conducted all-atom replica exchange molecular dynamics simulations.36,37 First, ss(GT)6 oligonucleotides were equilibrated on the (8,6) nanotube (Figure S7) to obtain an equilibrium configuration that exhibited a tight association between the ssDNA and nanotube (Figure 1d). Cholesterol molecules were then added and equilibrium was reached after 100 ns (Figure S7). In the resulting configuration, cholesterol bound to exposed regions on the nanotube and induced rearrangement of DNA on the nanotube surface (Figure S8). The combined effect was an 18.7% decrease in the density of water molecules within 1.2 nm of the nanotube surface (Figure 1e). These simulations were repeated with sphingomyelin molecules and a similar reduction in water density was observed (Figure 1d- e). The simulations suggest that lipid binding to the ss(GT)6-(8,6) complex reduces the water density on the nanotube surface, thereby lowering the effective local solvent dielectric. As experimentally observed, the lower dielectric environment corresponds to a blue shift of the nanotube emission wavelength (Figure 1c). We further characterized properties of the ss(GT)6-(8,6) optical response to cholesterol. The emission shift on cholesterol addition to surface-adsorbed complexes was rapid (under 2 minutes, limited by the hyperspectral instrument acquisition time, Figure S9). Sodium deoxycholate, a surfactant and water-soluble cholesterol analog, was added and subsequently removed from the surface-adsorbed complexes, demonstrating that the wavelength shift on analyte binding is intrinsically reversible (Figure S9). Furthermore, in an acidic environment, the response of the nanotube complex to lipids was similar to that at a neutral pH (Figure S9). Overall, the characteristics of the ss(GT)6-(8,6) complex suggest that it can function as a reporter of endolysosomal lipid accumulation in live cells. When prepared via previously described methods34 suspensions of ss(GT)6-(8,6) consist of short (~90 nm), singly dispersed nanotubes that are relatively free of impurities, and non-covalently functionalized with biocompatible single stranded DNA. This minimizes the key parameters of SWCNT cellular toxicity,30 a topic that is assessed below. Uptake and localization of DNA-SWCNTs to the endolysosomal lumen Although past work suggests that DNA-SWCNTs incubated with cells are taken up via energy dependent processes and localize to the endolysosomal lumen, this has not been assessed quantitatively or on a single-organelle level.39-41 We quantitatively assessed the interaction of ss(GT)6-(8,6) complexes with mammalian cells using near-IR and visible fluorescence microscopy. Macrophages were incubated in complete 10% FBS-supplemented cell culture media with 0.2 mg/L of ss(GT)6-(8,6) complexes at 37 °C for 30 minutes, before washing with fresh, complex-free media. For the conditions used in our experiments, this concentration corresponds to approximately 39 pM of ss(GT)6-(8,6).34 The cells exhibited bright near-IR emission, indicating that nanotubes were strongly associated with the cells (Figure S10). We observed that a ss(GT)6-(8,6) concentration of 0.2 mg/L, pulsed for 30 minutes, were the minima that resulted in sufficient NIR emission from all cell lines used in this work. We also measured the uptake of the ss(GT)6-(8,6) complexes as a function of temperature, and quantified the nanotube emission intensity associated with cells. Incubation at 4 °C resulted in 10-fold lower intensity than incubation at 37 °C (Figure S11), indicating that the complexes had been internalized by the cells via an energy dependent process. These results are consistent with previous reports of the energy-dependent uptake of ssDNA-nanotube complexes42 via endocytosis.39,40 The nanotube emission, quantified from over 700 individual cells incubated with the complex at 37 °C, followed a normal distribution, suggesting a relatively homogeneous uptake of the ss(GT)6-(8,6) complexes by the cells (Figure S10). To determine the localization of ss(GT)6-(8,6) complexes following uptake, we conducted a series of imaging experiments in macrophages (RAW 264.7 cell line and bone- marrow derived monocytes) and a human osteosarcoma cell line (U2OS cells). The ss(GT)6 oligonucleotide was covalently labeled with visible fluorophores (Cy3, Cy5, or Alexa-647) to prepare fluorophore-labeled ss(GT)6-nanotube complexes. Following internalization of 1 mg/L of the complexes by macrophages, we acquired emission from the Cy3 dye and NIR emission from the nanotubes in the same imaging field. This higher concentration of 1 mg/L was required to obtain sufficient emission; it was used for all experiments performed with the unsorted nanotube sample. The colocalization between the signals, observed on two different detectors, indicates that emission from a fluorophore conjugated to the ssDNA on the nanotube is a reliable indicator of nanotube location (Figure S12). Uptake and localization of DNA-SWCNTs to the endolysosomal lumen Next, we colocalized the Cy5-DNA-SWCNT fluorescent complexes with Lysotracker Green, a fluorescent probe that accumulates in endolysosomal organelles (Figure 2a, S13). A quantitative analysis using an unbiased auto- thresholding approach indicated significant colocalization between the Cy5 and Lysotracker Green emission (Pearson coefficient of 0.92 ± 0.036, Manders split colocalization coefficient of 0.95 ± 0.018), suggesting that the nanotube signal was contained within endolysosomal organelles. Concomitantly, the NIR emission from the nanotubes localized to the same regions of the cell as the visible emission from Lysotracker, further supporting the endolysosomal localization of the nanotubes (Figure S14). Figure 2. Localization of DNA-SWCNT to endolysosomal organelles. (a) Representative fluorescence microscopy images of 1 mg/L Cy5-labeled DNA-SWCNT complexes (red) and Lysotracker (green) in live cells. Scale bar = 10 µm. (b) Representative confocal images of TMR-dextran (green) and 1 mg/L Alexa-647-SWCNT (red) in U2OS-SRA cells. Lines (cyan) denote cross sections from the images extracted for further analysis in (c). Scale bar = 10 µm. (c) Intensity profiles of TMR (green) and Alexa 647 (red) fit with Gaussian functions. (d) Representative TEM images of AuNP-SWCNT complexes imaged on the TEM grid. Scale bar is 250 nm. (e) Representative TEM image of an AuNP-SWCNT complex within an endolysosomal organelle. Scale bar is 100 nm. (f) Relative frequency histogram of AuNP-SWCNT complexes per endolysosomal organelle. (g) Relative frequency histograms comparing the experimentally observed and predicted numbers of AuNPs per endolysosomal organelle if AuNPs were not attached to SWCNT complexes. Figure 2. Localization of DNA-SWCNT to endolysosomal organelles. (a) Representative fluorescence microscopy images of 1 mg/L Cy5-labeled DNA-SWCNT complexes (red) and Lysotracker (green) in live cells. Scale bar = 10 µm. (b) Representative confocal images of TMR-dextran (green) and 1 mg/L Alexa-647-SWCNT (red) in U2OS-SRA cells. Lines (cyan) denote cross sections from the images extracted for further analysis in (c). Scale bar = 10 µm. (c) Intensity profiles of TMR (green) and Alexa 647 (red) fit with Gaussian functions. (d) Representative TEM images of AuNP-SWCNT complexes imaged on the TEM grid. Scale bar is 250 nm. (e) Representative TEM image of an AuNP-SWCNT complex within an endolysosomal organelle. Scale bar is 100 nm. (f) Relative frequency histogram of AuNP-SWCNT complexes per endolysosomal organelle. (g) Relative frequency histograms comparing the experimentally observed and predicted numbers of AuNPs per endolysosomal organelle if AuNPs were not attached to SWCNT complexes. Uptake and localization of DNA-SWCNTs to the endolysosomal lumen In contrast, the experimentally determined number of endolysosomal organelles with two AuNP was four times higher (Figure 2g), suggesting that if two AuNP were observed within one endolysosomal organelle, then the two nanoparticles were statistically likely to be linked to each other via a nanotube. This, combined with the removal of free gold nanoparticles via centrifugation and the TEM images showing that all visualized AuNPs were attached to SWCNTs, suggests that the AuNPs within endolysosomal organelles were part of AuNP-SWCNT complexes. after removing free gold-nanotube complexes from the solution. In the cells, the gold nanoparticles were clearly visible as dark circles within endolysosomal organelles (Figure 2e, S16b-c). We quantified the number of AuNP-SWCNTs within each endolysosomal organelle (Figure 2f). From the relative frequency distribution, we found the probability that an endolysosomal organelle had one gold nanoparticle was 0.14. If two gold nanoparticles were to independently localize into the same vesicle, we calculated that the probability would be approximately 0.02 (0.14 x 0.14 = 0.02). In contrast, the experimentally determined number of endolysosomal organelles with two AuNP was four times higher (Figure 2g), suggesting that if two AuNP were observed within one endolysosomal organelle, then the two nanoparticles were statistically likely to be linked to each other via a nanotube. This, combined with the removal of free gold nanoparticles via centrifugation and the TEM images showing that all visualized AuNPs were attached to SWCNTs, suggests that the AuNPs within endolysosomal organelles were part of AuNP-SWCNT complexes. To determine the long-term fate of the complexes, we acquired NIR movies of ss(GT)6- (8,6) complexes within macrophages at 6, 24 and 48 hours after the initial 30 minute incubation (Supplementary Videos 1, 2, and 3). At each time point, emission from the complexes exhibited both passive diffusion and directed, linear movements consistent with the active translocation of lysosomes along microtubules,45 suggesting that the nanotube complexes remain within endolysosomal organelles. The emerging view, from our series of experiments, indicates the efficient uptake of DNA-nanotube complexes via endocytosis, rapid transport to the late endosomes and lysosomes, and stable localization to the lumen of endolysosomal organelles. Uptake and localization of DNA-SWCNTs to the endolysosomal lumen We next assessed the localization of the nanotubes within the lumen of individual endolysosomal organelles, by pulsing the cells with fluorescent (TMR) 10,000 MW polymer that accumulates in the endolysosomal lumen and does not degrade.43 Following overnight incubation, the cells were maintained in dextran-free media for 3 hours, before 1 mg/L Alexa-647 labeled nanotube complexes were introduced to the cell media for 30 minutes and then washed away. One hour later we performed high magnification confocal microscopy in the live cells. An analysis of over 40 cells (Figure 2b, S15) indicates that 50% TMR dextran labeled endolysosomal organelles colocalized with Alexa 647-nanotube complexes, suggesting that within an hour following their addition, the nanotubes had been transported to the dextran-loaded endolysosomal organelles. We extracted line intensity profiles of TMR and Alexa 647 emission and fit them with Gaussian functions (Figure 2c, S15). The single Gaussian intensity distributions of both fluorophores overlapped significantly, with centers that colocalized with diffraction-limited resolution. This result suggests that the nanotubes localized to the same region of endolysosomal organelles as dextran, which is known to remain in the endolysosomal lumen.43 To further confirm the presence of DNA-SWCNTs in the endolysosomal organelles, TEM analysis was performed. As single-walled carbon nanotubes, composed of only one layer of cylindrical graphene, do not have sufficient electron density to be visible by TEM in cells, we used gold-labeled DNA-nanotube complexes to perform the first incidence of gold-enhanced TEM imaging of individualized SWCNTs in mammalian cells. Citrate-capped gold nanoparticles (~ 10 nm diameter) were conjugated to thiolated ssDNA-nanotube complexes.44 Unbound gold nanoparticles were removed via centrifugation. Images of the gold nanoparticle-nanotube complexes (AuNP-SWCNT) deposited directly onto a TEM grid (Figure 2d, S16a) confirmed that all gold nanoparticles were attached to carbon nanotubes. We then incubated RAW 264.7 macrophages with 1 mg/L of the gold-labeled nanotubes, and fixed the cells for TEM imaging after removing free gold-nanotube complexes from the solution. In the cells, the gold nanoparticles were clearly visible as dark circles within endolysosomal organelles (Figure 2e, S16b-c). We quantified the number of AuNP-SWCNTs within each endolysosomal organelle (Figure 2f). From the relative frequency distribution, we found the probability that an endolysosomal organelle had one gold nanoparticle was 0.14. If two gold nanoparticles were to independently localize into the same vesicle, we calculated that the probability would be approximately 0.02 (0.14 x 0.14 = 0.02). Biocompatibility of DNA-SWCNT complexes in mammalian cells We conducted several experiments to assess the degree to which ss(GT)6-(8,6) perturbed cells and endolysosomal organelles in order to determine whether this may be a complication of its use as a live-cell sensor. Using an annexin V and propidium iodide assay, we found that, at its working concentration (0.2 mg/L), ss(GT)6-(8,6) did not affect cell viability or proliferation (Figure S17). To determine if DNA-SWCNT complexes altered the morphology of endolysosomal organelles, AuNP-SWCNT complexes were incubated with RAW 264.7 cells for 30 minutes before fixing and preparing for TEM imaging 6 hours later (Figure 3a). Analysis of the size, diameter and aspect ratio of endolysosomal organelles from the TEM images show no statistical differences between control macrophages and macrophages incubated with 1 mg/L complexes (Figure 3b, S18). Endolysosomal organelles in which gold-nanotubes were explicitly detected also displayed similar morphology to controls (Figure 3c, S19). At this elevated concentration of complexes (1 mg/L), we also did not observe a change in endolysosomal membrane permeabilization (Figure S20). We conducted several experiments to assess the degree to which ss(GT)6-(8,6) perturbed cells and endolysosomal organelles in order to determine whether this may be a complication of its use as a live-cell sensor. Using an annexin V and propidium iodide assay, we found that, at its working concentration (0.2 mg/L), ss(GT)6-(8,6) did not affect cell viability or proliferation We conducted several experiments to assess the degree to which ss(GT)6 (8,6) perturbed cells and endolysosomal organelles in order to determine whether this may be a complication of its use as a live-cell sensor. Using an annexin V and propidium iodide assay, we found that, at its working concentration (0.2 mg/L), ss(GT)6-(8,6) did not affect cell viability or proliferation (Figure S17). To determine if DNA-SWCNT complexes altered the morphology of endolysosomal organelles, AuNP-SWCNT complexes were incubated with RAW 264.7 cells for 30 minutes before fixing and preparing for TEM imaging 6 hours later (Figure 3a). Analysis of the size, diameter and aspect ratio of endolysosomal organelles from the TEM images show no (Figure S17). To determine if DNA-SWCNT complexes altered the morphology of endolysosomal organelles, AuNP-SWCNT complexes were incubated with RAW 264.7 cells for 30 minutes before fixing and preparing for TEM imaging 6 hours later (Figure 3a). Biocompatibility of DNA-SWCNT complexes in mammalian cells Analysis of the size, diameter and aspect ratio of endolysosomal organelles from the TEM images show no statistical differences between control macrophages and macrophages incubated with 1 mg/L complexes (Figure 3b, S18). Endolysosomal organelles in which gold-nanotubes were explicitly detected also displayed similar morphology to controls (Figure 3c, S19). At this elevated concentration of complexes (1 mg/L), we also did not observe a change in endolysosomal membrane permeabilization (Figure S20). Figure 3. Ultra structural analysis of endolysosomal organelles. (a) Representative TEM images of cells that were untreated or incubated with 1 mg/L of AuNP-SWCNT complexes. Endolysosomal organelles are shaded blue, scale bars are 2 µm. (b) A comparison of the mean aspect ratio (left) and area (right) of endolysosomal organelles. Error bars are standard deviation and mean values were compared with an unpaired t-test. (c) Histograms of the distribution of the aspect ratio, major axis, and area of endolysosomal organelles from control cells and endolysosomal organelles containing AuNP-SWCNT complexes. Figure 3. Ultra structural analysis of endolysosomal organelles. (a) Representative TEM ima tra structural analysis of endolysosomal organelles. (a) Representative TEM images of cells that Figure 3. Ultra structural analysis of endolysosomal organelles. (a) Representative TEM images of cells that were untreated or incubated with 1 mg/L of AuNP-SWCNT complexes. Endolysosomal organelles are shaded blue, scale bars are 2 µm. (b) A comparison of the mean aspect ratio (left) and area (right) of endolysosomal organelles. Error bars are standard deviation and mean values were compared with an unpaired t-test. (c) Histograms of the distribution of the aspect ratio, major axis, and area of endolysosomal organelles from control cells and endolysosomal organelles containing AuNP-SWCNT complexes. We next assessed whether DNA-SWCNTs perturbed the ability of endolysosomal organelles to maintain their pH gradient. This was done via a confocal imaging study using LysoTracker, a lysomotropic fluorescent probe that accumulates in acidic vesicles. Human bone osteosarcoma cells transfected with type A scavenger receptors (U2OS-SRA)46 were treated with 1 mg/L (five times higher than the working concentration) fluorophore-labeled DNA-nanotube complexes (Alexa 647-SWCNT) and fixed and stained with LysoTracker. Using fluorescence confocal microscopy, we imaged both Lysotracker and Alexa 647-SWCNT emission from endolysosomal organelles (Figure 4a, S21). If DNA-SWCNTs prevented endolysosomal organelles from maintaining a pH gradient, we would expect endolysosomal organelles containing the DNA-SWCNTs to show decreased levels of LysoTracker signal. Biocompatibility of DNA-SWCNT complexes in mammalian cells Moreover, as electronic structure and chirality of a nanotube sample were not found to directly affect toxological impact,47 our of a nanotube sample were not found to directly affect toxological impact,47 our conclusions on biocompatibility likely hold for other relatively short DNA-single walled carbon nanotube complexes composed of different DNA sequences and chiralities. Figure 4. Assessing the effects of DNA-SWCNT on endolysosomal function. (a) Representative confocal images of LysoTracker (green), Alexa 647-SWCNT (red) and a merged image of the two in U2OS-SRA cells Scale bar is 10 µm. (b) Intensity profiles of the two fluorophores along the dashed line (cyan) in the merged image. (c) Mean intensity of LysoTracker in endolysosomal organelles that contain Alexa 647-SWCNT emission and those that did not. Error bars are standard deviation. Mean intensity was compared with an unpaired t-test (n=9 images per channel). (d) Representative epifluorescence images of Alexa 488-AcLDL (green) in U2OS-SRA cells, at 0 and 2 hours after the addition of acetylated LDL, or in control cells. Scale bars are 10 µm. (e) Number of AcLDL ROIs per 100 µm2. Error bars are standard deviation, obtained from 10 images per condition. Data were compared using a one way ANOVA with Tukey’s multiple comparison test. ing the effects of DNA-SWCNT on endolysosomal function. (a) Representative confocal images Figure 4. Assessing the effects of DNA-SWCNT on endolysosomal function. (a) Representative confocal images of LysoTracker (green), Alexa 647-SWCNT (red) and a merged image of the two in U2OS-SRA cells Scale bar is 10 µm. (b) Intensity profiles of the two fluorophores along the dashed line (cyan) in the merged image. (c) Mean intensity of LysoTracker in endolysosomal organelles that contain Alexa 647-SWCNT emission and those that did not. Error bars are standard deviation. Mean intensity was compared with an unpaired t-test (n=9 images per channel). (d) Representative epifluorescence images of Alexa 488-AcLDL (green) in U2OS-SRA cells, at 0 and 2 hours after the addition of acetylated LDL, or in control cells. Scale bars are 10 µm. (e) Number of AcLDL ROIs per 100 µm2. Error bars are standard deviation, obtained from 10 images per condition. Data were compared using a one way ANOVA with Tukey’s multiple comparison test. Biocompatibility of DNA-SWCNT complexes in mammalian cells This was not the case as quantification of LysoTracker fluorescence from organelles both with and without DNA- SWNCT showed no significant difference in LysoTracker intensity (Figure 4b-c, S21). To ensure that we could ascertain meaningful results from a DNA-SWCNT based reporter, we also investigated the effect of DNA-SWCNTs on the ability of endolysosomal organelles to hydrolyze lipoprotein molecules. We treated U2OS-SRA cells with 1 mg/L of Alexa 647-SWCNT for 30 minutes before incubation in fresh media for two hours. To induce the rapid uptake of lipoproteins, we then introduced 50 µg/mL of Alexa 488-labeled acetylated LDL (Alexa 488-AcLDL) to the cell media. Epifluorescence images showed a stark decrease in AcLDL puncta in both control and nanotube-treated cells between zero and two hours of AcLDL addition (Figure 4d, S22). Quantification of AcLDL regions of interest (ROIs), normalized by cell area showed that there was no significant difference in the number of AcLDL ROIs per 100 µm2 between cells treated with the nanotubes and the control cells at both 0 and 2 hours (Figure 4e), suggesting that DNA-SWCNTs did not perturb the hydrolysis of lipoproteins. A lipid biochemical assay showed no significant differences in the cholesterol or total lipid content of cell fractions in control and nanotube-containing cells (Figure S23). LipidTox imaging and the expression of LDL receptor (LDLr), also suggested that DNA-SWCNT complexes did not significantly alter lipid processing in cells (Figure S24-25). The above results suggest that singly-dispersed DNA-SWCNTs that have been separated from large nanotube bundles and carbonaceous impurities via ultra-centrifugation did not adversely affect cell viability or proliferation, or organelle morphology, integrity, or capacity to digest lipoproteins. However, additional work using different cell types and culture conditions are warranted prior to widespread application of the reporter. Moreover, as electronic structure and chirality of a nanotube sample were not found to directly affect toxological impact,47 our conclusions on biocompatibility likely hold for other relatively short DNA-single walled carbon nanotube complexes composed of different DNA sequences and chiralities. warranted prior to widespread application of the reporter. Moreover, as electronic structure and chirality of a nanotube sample were not found to directly affect toxological impact,47 our conclusions on biocompatibility likely hold for other relatively short DNA-single walled carbon warranted prior to widespread application of the reporter. Detecting lipid accumulation in the endolysosomal lumen of live cells We investigated whether the ability of ss(GT)6 –(8,6) to detect lipids in vitro could be translated to the endolysosomal organelles of live cells (Figure 5a). RAW 264.7 macrophages were incubated with the complexes for 30 minutes in complete 10% serum-supplemented media at 37° C and washed with fresh media. To induce lysosomal accumulation of free cholesterol, we prepared cells that were pre-treated with U18666A,48 a compound that inhibits the action of NPC1,49 a membrane protein that effluxes free cholesterol out of the lysosomes (Figure 5a), to mimic the Niemann-Pick C1 disease phenotype. Cells were also pre-treated with Lalistat 3a2,50 an inhibitor of lysosomal acid lipase (LAL), which is the central enzyme that hydrolyzes low density lipoproteins (Figure 5a).51 Inhibition of LAL leads to the accumulation of esterified cholesterol, which is observed in Wolman’s disease. Using near-IR hyperspectral microscopy, we acquired the emission from the complexes localized within the endolysosomal organelles of macrophages under these three conditions (Figure 5a). When the emission wavelength is mapped to a color-scale and overlaid on a transmitted light image, the spatially-resolved emission from ss(GT)6-(8,6) complexes resulted in live-cell maps of endolysosomal lipid content (Figure 5b). The mean emission blue-shift for the two drug-treated conditions was over 11 nm compared with the control, with a single population for all three conditions, indicating lipid accumulation in all endolysosomal organelles that contained the complexes (Figure 5c, S26). Neither Lalistat 3a2 nor U18666A directly perturbed the emission wavelength of the complexes (Figure S27). The endolysosomal lipid maps thus reflect the optical response of the complexes to the accumulation of lipids within the endolysosomal lumen. We quantified the nanotube emission response as a function of loading concentration. At a 5-fold dilution of the working concentration, the response of the emission wavelength to U18666A-induced lipid accumulation did not change, indicating that the emission spectra (Figure S28-29) and wavelength distribution of the ss(GT)6-(8,6) complex (Figure S28- 29) was consistent over a range of concentrations (Figure S28-29). Finally, we induced endolysosomal lipid accumulation in mouse embryonic fibroblasts (MEF) using U18666A, and detected a blue-shift in the ss(GT)6-(8,6) emission when compared with untreated controls (Figure S30). Based on the findings from our experiments, we present a model for the optical response of ss(GT)6-(8,6) ssDNA-nanotube complexes in live cells under the conditions investigated. The complexes enter the endolysosomal pathway and localize within the lumen of endolysosomal organelles. Detecting lipid accumulation in the endolysosomal lumen of live cells In the complex environment of the endolysosomal lumen, the nanotube emission functions as a cell-specific spectral fingerprint for the lipid content. Upon dysfunction of the NPC1 protein via introduction of U18666A to cells, free cholesterol (FC) accumulates within the lumen of endolysosomal organelles and adsorbs to the surface of the ss(GT)6-(8,6) complex, resulting in a distinct blue-shifting response of the nanotube emission. Similarly, upon introduction of Lalistat to cells, cholesteryl esters (CE) accumulate in the endolysosomal lumen, resulting in the adsorption of CE to the nanotube surface, and a concomitant blue-shifting response. In both situations, multiple secondary lipids also accumulate in the lysosomal lumen.52 Hence, we find that the ss(GT)6-(8,6) DNA-nanotube complex functions as a quantitative optical reporter of lipid accumulation in the endolysosomal lumen via solvatochromic shifting of intrinsic carbon nanotube photoluminescence. Henceforth, we refer to ss(GT)6-(8,6) as the ‘reporter’. Based on the findings from our experiments, we present a model for the optical response of ss(GT)6-(8,6) ssDNA-nanotube complexes in live cells under the conditions investigated. The complexes enter the endolysosomal pathway and localize within the lumen of endolysosomal organelles. In the complex environment of the endolysosomal lumen, the nanotube emission functions as a cell-specific spectral fingerprint for the lipid content. Upon dysfunction of the NPC1 protein via introduction of U18666A to cells, free cholesterol (FC) accumulates within the lumen of endolysosomal organelles and adsorbs to the surface of the ss(GT)6-(8,6) complex, As the nanotube emission is exclusively from the endolysosomal lumen, we investigated whether the nanotube spectra alone, obtained without imaging, could function as an analytical tool to benchmark drug-induced endolysosomal lipid accumulation in a high-throughput format. Using a customized NIR spectrometer,53 we obtained spectra from complexes within live RAW 264.7 macrophages (Figure 5d) and detected a 9 nm shift in the emission from cells treated with U18666A –an example of a cationic amphiphilic drug that induces DIPL54 (Figure 5e). These results were similar to the hyperspectral data of control and U18666A-treated macrophages (Figure 5f), indicating the amenability of the complexes for both imaging and a spectroscopy- based assay which facilitates higher throughput. Figure 5. Detection of endolysosomal lipid accumulation in live cells. (a) Schematics of the ss(GT)6-(8,6) nanotube complexes in macrophages treated with U18666A or Lalistat. (b) Overlay of transmitted light with hyperspectral image of RAW 264.7 macrophages incubated with ss(GT)6-(8,6) complex under the specified treatments. Color legend maps to nanotube emission peak wavelength. Detecting lipid accumulation in the endolysosomal lumen of live cells Scale bar = 50 µm. (c) Histogram of emission wavelengths of all pixels from the hyperspectral images, bin size of 1 nm. (d) Schematic of the optical setup for high-throughput measurements of ss(GT)6-(8,6) emission in live cells. (e) Spectra of ss(GT)6-(8,6) emission from live RAW 264.7 cells incubated in normal media (control), and in media with U18666A. (f) Mean center Figure 5. Detection of endolysosomal lipid accumulation in live cells. (a) Schematics of the ss(GT)6-(8,6) nanotube complexes in macrophages treated with U18666A or Lalistat. (b) Overlay of transmitted light with hyperspectral image of RAW 264.7 macrophages incubated with ss(GT)6-(8,6) complex under the specified treatments. Color legend maps to nanotube emission peak wavelength. Scale bar = 50 µm. (c) Histogram of emission wavelengths of all pixels from the hyperspectral images, bin size of 1 nm. (d) Schematic of the optical setup for high-throughput measurements of ss(GT)6-(8,6) emission in live cells. (e) Spectra of ss(GT)6-(8,6) emission from live RAW 264.7 cells incubated in normal media (control), and in media with U18666A. (f) Mean center wavelengths of ss(GT)6-(8,6) emission from n = 5 technical replicates, acquired using hyperspectral imaging (I) and spectroscopy (S). Mean values were compared using a one way ANOVA with Sidak’s multiple comparison test. Error bars are standard error of the mean. =p<.001, =p<.0001. wavelengths of ss(GT)6-(8,6) emission from n = 5 technical replicates, acquired using hyperspectral imaging (I) and spectroscopy (S). Mean values were compared using a one way ANOVA with Sidak’s multiple comparison test. Error bars are standard error of the mean. =p<.001, **=p<.0001. fibroblasts. The reporter was assessed for its response in primary cells from a patient with Niemann- Pick type C1 (NPC1), a lysosomal storage disease characterized by an accumulation of unesterified cholesterol in the lysosomes.55,56 Fibroblasts from an NPC patient, as well as wild- type (WT) human fibroblasts, were incubated with the reporter for 30 minutes, then rinsed, and incubated in fresh media. Hyperspectral data collected after 24 hours indicates that the reporter blue-shifted by an average of 6 nm in NPC1 human fibroblasts, compared to WT human fibroblasts (Figure 6a). The lipid content of WT fibroblast endolysosomal organelles was relatively homogeneous, as evinced by the narrow distribution of the reporter emission (Figure 6b). In contrast, the reporter exhibited a broad emission profile within NPC1 cells (Figure 6b), confirming the published findings that NPC1 cells exhibit a wide distribution of lipid concentrations.57 The histogram also suggests that a sizeable fraction of endolysosomal organelles contain near-normal cholesterol content. To test the reversibility of the reporter in live cells, we measured the emission from the reporter in NPC1 fibroblasts treated with hydroxypropyl-β-cyclodextrin (HPβCD), a drug known to facilitate the efflux of accumulated cholesterol from the lumen of endolysosomal organelles.58 After treatment with 100 µM HPβCD for 24 hours, hyperspectral imaging revealed significant red-shifting of the reporter emission from the same NPC1 cells that were previously blue-shifted (Figure 6a-b, S31). The distribution of reporter emission from the HPβCD-treated NPC1 fibroblasts appeared notably similar to the emission from WT fibroblasts (Figure 6b, S31), and reduction of total cell cholesterol content was confirmed by fixing the cells and staining with filipin (Figure 6c). Filipin staining was more pronounced in NPC1 fibroblasts as compared to WT fibroblasts, and HPβCD treatment resulted in significantly diminished filipin signal in NPC1 cells (Figure 6c, S32), thus validating the results obtained from the reporter alone. Additionally, pre-treatment of NPC1 fibroblasts with HPβCD prevented the blue-shifting of the reporter (Figure S33). Figure 6. Measurement of endolysosomal lipid accumulation and reversal in NPC1 patient-derived fibroblasts. (a) Mean reporter emission from wild-type fibroblasts, patient-derived NPC1 fibroblasts, and NPC1 fibroblasts treated with hydroxypropyl-β-cyclodextrin (HPβCD) for 24 hours. Statistical significance was determined with a one-way ANOVA with Sidak’s multiple comparison test. (b) Histograms of the nanotube emission wavelength from single endolysosomal organelles of wild-type fibroblasts, NPC1 patient fibroblasts, and NPC1 fibroblasts treated with HPβCD for 24 hours. fibroblasts. (c) Mean filipin intensity from WT fibroblasts, NPC1 cells, and NPC1 cells treated with HPβCD for 24 hours, at 48 hours after nanotube addition. Statistical significance was determined with a one-way ANOVA with Tukey’s multiple comparison test. All error bars are S.E.M. from 3 technical replicate experiments. = p<0.05, = p<0.01, * = p<0.001. Figure 6. Measurement of endolysosomal lipid accumulation and reversal in NPC1 patient-derived fibroblasts. (a) Mean reporter emission from wild-type fibroblasts, patient-derived NPC1 fibroblasts, and NPC1 fibroblasts treated with hydroxypropyl-β-cyclodextrin (HPβCD) for 24 hours. Statistical significance was determined with a one-way ANOVA with Sidak’s multiple comparison test. (b) Histograms of the nanotube emission wavelength from single endolysosomal organelles of wild-type fibroblasts, NPC1 patient fibroblasts, and NPC1 fibroblasts treated with HPβCD for 24 hours. (c) Mean filipin intensity from WT fibroblasts, NPC1 cells, and NPC1 cells treated with HPβCD for 24 hours, at 48 hours after nanotube addition. Statistical significance was determined with a one-way ANOVA with Tukey’s multiple comparison test. All error bars are S.E.M. from 3 technical replicate experiments. * = p<0.05, = p<0.01, * = p<0.001. Single-cell kinetic measurements of lipid accumulation Recent technological advances have lead to an appreciation of the complexity of cell populations and the heterogeneity of single cells within a population of cells that seems uniform on a macroscopic level.59 The ability of the developed reporter to function in live cell imaging applications positions it to provide data on a single cell level. Such data would have the potential to identify previously unknown sub-populations of cells that could be crucial towards increasing our understanding of cellular lipid processing. We assessed whether the reporter could measure single-cell kinetics of endolysosomal lipid accumulation. RAW 264.7 macrophages were cultured in media with lipoprotein depleted serum (LPDS). Under these conditions the reporter emission wavelength was approximately 1200 nm. Next, both AcLDL and U18666A were added to the cell media to induce endolysosomal lipid accumulation. Endolysosomal lipid accumulation was monitored via the acquisition of hyperspectral images every 10 minutes for 2 hours (Figure 7a). Single-cell emission trajectories were computed for all cells with reporter peak emission intensities of over 4 times higher than the background. The mean reporter emission from individual cells blue-shifted and reached equilibrium over approximately 90 minutes (Figure 7b). For each cell, the spectral trajectories were fit with a single exponential function to obtain the time constant, time lag, and the starting and final reporter wavelengths (Figure S34). The time constants of lipid accumulation in the cells averaged approximately 40 minutes and followed a lognormal distribution (Figure 7c). One potential explanation for this finding is that a lognormal distribution is often observed for a quantity arising from multiple serial processes.60 We believe that this distribution of time constants from individual cells is consistent with the process of endolysosomal lipid accumulation, which involves sequential processes including LDL uptake We assessed whether the reporter could measure single-cell kinetics of endolysosomal lipid accumulation. RAW 264.7 macrophages were cultured in media with lipoprotein depleted serum (LPDS). Under these conditions the reporter emission wavelength was approximately 1200 nm. Next, both AcLDL and U18666A were added to the cell media to induce 1200 nm. Next, both AcLDL and U18666A were added to the cell media to induce endolysosomal lipid accumulation. Endolysosomal lipid accumulation was monitored via the acquisition of hyperspectral images every 10 minutes for 2 hours (Figure 7a). Single-cell emission trajectories were computed for all cells with reporter peak emission intensities of over 4 times higher than the background. Single-cell kinetic measurements of lipid accumulation The mean reporter emission from individual cells blue-shifted and reached equilibrium over approximately 90 minutes (Figure 7b). For each cell, the spectral trajectories were fit with a single exponential function to obtain the time constant, time lag, and the starting and final reporter wavelengths (Figure S34). The time constants of lipid accumulation in the cells averaged approximately 40 minutes and followed a lognormal distribution (Figure 7c). One potential explanation for this finding is that a lognormal distribution endolysosomal lipid accumulation, which involves sequential processes including LDL uptake into the early endosome, delivery of LDL to the lysosome, hydrolysis of esterified cholesterol by LAL, and the inhibited efflux of free cholesterol by NPC1. into the early endosome, delivery of LDL to the lysosome, hydrolysis of esterified cholesterol by LAL, and the inhibited efflux of free cholesterol by NPC1. Interestingly, the distribution of time constants showed marked heterogeneity, with the slowest and fastest time constants differing by an order of magnitude. Cells exhibiting slower rates of lipid accumulation also exhibited an initial state of relative lipid deficiency. The time constant of lipid accumulation modestly correlated with the starting wavelength (Spearman correlation of 0.33, p<0.01). Notably, the slowest shifting cells (defined as cells with a time constant > 90 minutes) exhibited an initial average emission wavelength >1202 nm, 2 nm more red-shifted than the faster shifting cells (Figure 7d). This result suggests the existence of a subpopulation of cells that maintained both lipid-deficient endolysosomal organelles and an extremely slow rate of lipid accumulation; i.e. these individual cells in the population may be especially resistant to endolysosomal lipid accumulation. Figure 7. Single-cell kinetics of lipid accumulation. (a) Overlaid brightfield and hyperspectral images of the reporter emission in RAW 264.7 macrophages upon addition of AcLDL (100 µg/mL) and U18666A (3 µg/mL) to LPDS media at t = 0, 30 minutes, and 60 minutes. Scale bar = 50 µm. (b) Single-cell trajectories of lysosomal lipid accumulation from 60 cells. Black curve is the mean. Error bars are standard deviation from n = 4 technical replicates. (c) Distribution of time constants of lipid accumulation in single cells fitted to a log-normal distribution. Bin size = 20 nm. (d) Scatter plot of the time constants vs. starting emission wavelength, n = 60 cells. Data from 4 independent experiments were combined for this analysis. Figure 7. Single-cell kinetics of lipid accumulation. Single-cell kinetic measurements of lipid accumulation (a) Overlaid brightfield and hyperspectral images of the reporter emission in RAW 264.7 macrophages upon addition of AcLDL (100 µg/mL) and U18666A (3 µg/mL) to LPDS media at t = 0, 30 minutes, and 60 minutes. Scale bar = 50 µm. (b) Single-cell trajectories of lysosomal lipid accumulation from 60 cells. Black curve is the mean. Error bars are standard deviation from n = 4 technical replicates. (c) Distribution of time constants of lipid accumulation in single cells fitted to a log-normal distribution. Bin size = 20 nm. (d) Scatter plot of the time constants vs. starting emission wavelength, n = 60 cells. Data from 4 independent experiments were combined for this analysis. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles After demonstrating the reporter’s ability to quantify lipid accumulation on a single cell level, we investigated whether the reporter could measure lipids on single-organelle level. We generated hyperspectral maps to quantify the intracellular heterogeneity of lipid content within individual endolysosomal organelles of single bone marrow derived monocytes during the differentiation process into bone marrow derived macrophages (BMDM) (Figure 8a). After demonstrating the reporter’s ability to quantify lipid accumulation on a single cell level, we investigated whether the reporter could measure lipids on single-organelle level. We generated hyperspectral maps to quantify the intracellular heterogeneity of lipid content within individual endolysosomal organelles of single bone marrow derived monocytes during the differentiation process into bone marrow derived macrophages (BMDM) (Figure 8a). Hyperspectral images of two cells (Figure 8b) from the bone marrow-derived monocyte population on day 3 of differentiation showed distinctly different intracellular heterogeneity of the reporter response, evident from the histogram widths (Figures 8c, S35). To mathematically quantify the intracellular heterogeneity of reporter emission within each cell, we used the normalized Simpson’s Index (nSI), a statistical measure of diversity.61,62 For the cells shown in Figure 8c, the nSIs were significantly different (Cell 1 nSI = 0.18, Cell 2 nSI = 0.74). By plotting the mean reporter emission wavelength and nSI for a large population of cells on day 3, cells with the most lipid-rich endolysosomal organelles (shorter wavelength) exhibited greater intracellular heterogeneity in endolysosomal lipid contents (Figure 8d, Spearman correlation of 0.33, p < 0.01, for n = 64 cells). This observation, confirmed in BMDMs isolated from 4 different mice (Figure S36, with at least 60 cells analyzed per mouse), suggests that within differentiating monocytes, cells with lipid-rich endolysosomal organelles maintain a sub- population of these organelles that are relatively lipid-deficient. Moreover, observation of BMDMs throughout differentiation validated the ability of the reporter to detect endolysosomal lipid accumulation in a non-pathological condition (Figure S37-38). Figure 8. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles. (a) Near-infrared broadband and overlaid brightfield/hyperspectral images of the reporter in bone marrow-derived cells after 3 days of colony stimulating factor-1 (CSF-1) treatment. (b) Endolysosomal lipid maps of the reporter in bone marrow- derived cells at day 3 of maturation. (c) Corresponding histogram of reporter emission from emissive pixels within the two cells. Bin size = 1 nm. (d) Scatterplot of the normalized Simpson’s Index (nSI) against the mean emission wavelength per cell for n = 64 cells. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles Scale bars = 10 µm Figure 8. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles. (a ifying intracellular heterogeneity in monocyte endolysosomal organelles. (a) Near-infrared Figure 8. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles. (a) Near-infrared broadband and overlaid brightfield/hyperspectral images of the reporter in bone marrow-derived cells after 3 days of colony stimulating factor-1 (CSF-1) treatment. (b) Endolysosomal lipid maps of the reporter in bone marrow- derived cells at day 3 of maturation. (c) Corresponding histogram of reporter emission from emissive pixels within the two cells. Bin size = 1 nm. (d) Scatterplot of the normalized Simpson’s Index (nSI) against the mean emission wavelength per cell for n = 64 cells. Scale bars = 10 µm Figure 8. Quantifying intracellular heterogeneity in monocyte endolysosomal organelles. (a) Near-infrared broadband and overlaid brightfield/hyperspectral images of the reporter in bone marrow-derived cells after 3 days of colony stimulating factor-1 (CSF-1) treatment. (b) Endolysosomal lipid maps of the reporter in bone marrow- derived cells at day 3 of maturation. (c) Corresponding histogram of reporter emission from emissive pixels within the two cells. Bin size = 1 nm. (d) Scatterplot of the normalized Simpson’s Index (nSI) against the mean emission wavelength per cell for n = 64 cells. Scale bars = 10 µm DNA encapsulation of single-walled carbon nanotubes The chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO, US) and Fisher Scientific (Pittsburgh, US). Single-walled carbon nanotubes (SWCNTs) produced by the HiPco process were used throughout the study (Unidym, Sunnyvale, CA). The carbon nanotubes were dispersed with DNA oligonucleotides via probe-tip ultrasonication (Sonics & Materials, Inc.) of 2 mg of the specified oligonucleotide (IDT DNA, Coralville, IA) with 1 mg of raw SWCNT in 1 mL of 0.1 M NaCl for 30 minutes at 40% of the maximum amplitude of the ultrasonicator (SONICS Vibra Cell). Following ultrasonication, the dispersions were ultracentrifuged (Sorvall Discovery 90SE) for 30 minutes at 280,000 x g. The top ¾ of the resultant supernatant was collected and its concentration was determined with a UV/Vis/NIR spectrophotometer (Jasco, Tokyo, Japan) using the extinction coefficient Abs910 = 0.02554 L mg-1cm-1 19. To remove free DNA, 100 kDa Amicon centrifuge filters (Millipore) were used to concentrate and re-suspend the DNA-nanotube complexes. CONCLUSION In this work, we developed a carbon nanotube optical reporter, composed of the (8,6) SWCNT species non-covalently functionalized with a short DNA oligonucleotide, that can function as an optical reporter of lipid content within the endolysosomal lumen of live cells. The near-infrared photoluminescence of the nanotube responds quantitatively to lipids in the local environment of the reporter via shifting of the nanotube optical bandgap. Experimental evidence and all-atom molecular dynamics simulations suggest that the mechanism of the response is solvatochromic shifting due to the reduction in water density on the nanotube surface. The reporter remains within the endocytic pathway and localizes to the lumen of endolysosomal organelles without adversely affecting organelle morphology, structural integrity, or function. In the endolysosomal lumen, the reporter’s near-infrared emission responds rapidly and reversibly to lipid accumulation. Via NIR hyperspectral microscopy, the reporter can quantitatively map lipids in live cells. Using spectroscopy, the reporter can measure endolysosomal lipid accumulation in live cells in a high-throughput drug screening-type format. The reporter detected lipid accumulation in lysosomal storage disorders, including in the endolysosomal organelles of fibroblasts derived from a patient with Niemann-Pick type C, as well as phenotypic reversal in the same cells after drug treatment. The reporter functioned with single-cell and single-organelle resolution and was used to assess single-cell kinetics of modified LDL accumulation within endolysosomal organelles, showing that the rate of cholesterol accumulation differs by an order of magnitude across macrophages in the same population of cells. Endolysosomal lipid accumulation in differentiating bone marrow-derived monocytes was also observed, and high resolution endolysosomal lipid maps revealed intracellular heterogeneity in the form of a sub- population of lipid-deficient endolysosomal organelles in lipid-rich cells. As the first technique for measuring lipid flux in the endolysosomal lumen of live cells, we expect this tool will have broad utility in both drug screening applications and in the investigation of disease pathways associated with altered lipid biology such as atherosclerosis, neurodegenerative diseases, lysosomal storage disorders, and liver disease. and all-atom molecular dynamics simulations suggest that the mechanism of the response is solvatochromic shifting due to the reduction in water density on the nanotube surface. The reporter remains within the endocytic pathway and localizes to the lumen of endolysosomal organelles without adversely affecting organelle morphology, structural integrity, or function. In the endolysosomal lumen, the reporter’s near-infrared emission responds rapidly and reversibly to lipid accumulation. Via NIR hyperspectral microscopy, the reporter can quantitatively map Purification of single-chirality nanotube complexes Carbon nanotubes were separated by an ion-exchange chromatography (IEX) method according to the procedure described by Tu et al.32 Briefly, unsorted HiPco nanotubes were dispersed using a DNA oligonucleotide with the sequence ss(GT)6, as described above. The sample was injected into a high performance liquid chromatograph (HPLC) (Agilent, 1260 Infinity) fitted with an anion-exchange column (Biochrom Labs, Inc., CNT-NS1500) with a running buffer of 2x SSC at a flow rate of 2 mL/min. A linearly increasing salt concentration gradient of 1M NaSCN (5%/min) was used to elute the nanotubes from the stationary phase and fractions were collected. The first fraction exciting the HPLC contained the highest purity of the (8,6) species, estimated at 86%, which was used for subsequent studies. The first fraction exciting the HPLC contained the highest purity of the (8,6) species, estimated at 86%, which was used for subsequent studies. Near-infrared fluorescence microscopy of single-walled carbon nanotubes As described in a previous study,19 near infrared fluorescence microscopy was used to acquire the photoluminescence emission from SWCNTs. The system comprised a continuous wave (CW) 730 nm diode laser with an output power of 2 W injected into a multimode fiber to produce the excitation source for fluorescence experiments. To ensure a homogenous illumination over the entire microscope field of view, the excitation beam passed through a custom beam-shaping module to produce a top-hat intensity profile with under 20% power variation on the imaged region of the sample. The final power at the sample was 230 mW. A long pass dichroic mirror with a cut-on wavelength of 875 nm (Semrock) was aligned to reflect the laser to the sample stage of an Olympus IX-71 inverted microscope (with internal optics modified to improve near-infrared transmission from 900-1400 nm) equipped with a 20X LCPlan N, 20X/0.45 IR objective and a UAPON100XOTIRF, 1.49 oil objective (Olympus, USA). Emission was collected with a 2D InGaAs array detector (Photon Etc.). Z-stacks of broadband, widefield emission (from 900 to 1650 nm) were acquired in steps of 0.5 µm, which satisfied the Nyquist sampling criterion for 100X magnification. Custom codes, written using Matlab software, were used to subtract background, correct for non-uniformities in excitation profile, and compensate for dead pixels on the detector. Hyperspectral microscopy was conducted by passing the emission through a volume Bragg grating (VBG) placed immediately before the InGaAs array in the optical path. The filtered image produced on the InGaAs camera was composed of a series of vertical lines, each with a specific wavelength. The reconstruction of a spatially-rectified image stack was performed using cubic interpolation on every pixel for each monochromatic image, according to the wavelength calibration parameters. The rectification produced a hyperspectral “cube” of images of the same spatial region exhibiting distinct spectral regions with 3.7 nm FHWM bandwidths. Approximately 50% of the emission intensity from the nanotubes is reduced after passage through the VBG. Analysis and processing of hyperspectral data Hyperspectral data acquired was saved as a (320 x 256 x 26) 16-bit array, where the first two coordinates signify the spatial location of a pixel and the last coordinate is its position in wavelength space. For the (8,6) nanotube, the 26-frame wavelength space ranges from 1150 nm to 1250 nm. An initial filter removed any pixels with a maximum intensity value outside (1170 to 1220 nm), as these were background pixels that emit outside the range for (8,6). For the remaining pixels, a peak-finding algorithm was used to calculate the intensity range for a given pixel i.e. range = (intensity_maximum – intensity_minimum). A data point was designated as a peak if its intensity was range/4 greater than the intensity of adjacent pixels. Pixels that failed the peak-finding threshold, primarily due to low intensity above the background, were removed from the data sets. The remaining pixels were fit with a Lorentzian function. Preparation of gold nanoparticle-conjugated nanotubes Gold nanoparticle-conjugated nanotubes were prepared according to a previously published study. Briefly, 10 nm citrate-capped gold nanoparticles were synthesized by bringing 50 mL of 0.01 wt% HAuCl4 and adding 2 mL of 1 wt% Sodium(III) citrate. After 2 minutes, the solution turns bright red, indicating nanoparticle formation. The gold nanoparticles were stabilized via a ligand exchange reaction by shaking overnight with bis(p-sulfonatophenyl)phenyl phosphine dihydrate dipotassium salt. The nanoparticles were then centrifuged and resuspended in deionized water. In parallel, ss(GT)27-T6-thiol-dispersed HiPco nanotube complexes were created by means of the previously described sonication and centrifugation protocol. The nanotube complexes were filtered with ultracentrifuge filters three times to remove unbound DNA. The nanotube complexes and excess gold nanoparticles were then shaken overnight. The unbound gold nanoparticles were removed via centrifugation, which would pellet the unbound nanoparticles, and careful supernatant extraction. Preparation of nanotubes labeled with visible fluorophores To increase the fluorescence intensity of the Cy3 or Cy5 fluorophores attached to DNA strands encapsulating SWCNTs, a 6 nucleotide long polyT tail was added to the end of the (GT)6 To increase the fluorescence intensity of the Cy3 or Cy5 fluorophores attached to DNA strands encapsulating SWCNTs, a 6 nucleotide long polyT tail was added to the end of the (GT)6 sequence, as fluorophores near the surface of SWCNTs are known to quench63 (Integrated DNA Technologies, sequence = GTGTGTGTGTGTTTTTTT). For confocal imaging with Alexa-647 sequence, as fluorophores near the surface of SWCNTs are known to quench63 (Integrated DNA Technologies, sequence = GTGTGTGTGTGTTTTTTT). For confocal imaging with Alexa-647 SWCNT, a small polyethylene glycol spacer was also added to further increase the fluorescence intensity of the fluorophore (Integrated DNA Technologies, sequence = GTGTGTGTGTGTTTTTTT/iSP18//3Alexaf647N//3’). These modified DNA-strands were non- covalently complexed with HiPco SWCNTs via the previously described sonication and centrifugation protocol. SWCNT, a small polyethylene glycol spacer was also added to further increase the fluorescence intensity of the fluorophore (Integrated DNA Technologies, sequence = Transmission electron microscopy (TEM) imaging Gold nanoparticle-nanotube conjugates were first imaged on carbon coated TEM grids by letting a 20 µL drop evaporate in the center of the grid. For imaging in RAW 264.7 cells, gold nanoparticle-nanotubes were introduced to the media for 30 minutes at 1 mg/L, and then washed thoroughly and replaced with fresh media. After 6 hours, cells were washed with serum-free media then fixed with a modified Karmovsky's fix of 2.5% glutaraldehyde, 4% paraformaldehyde and 0.02% picric acid in 0.1M sodium cacodylate buffer at pH 7.2. Following a secondary fixation in 1% osmium tetroxide and 1.5% potassium ferricyanide, samples were dehydrated through a graded ethanol series, and embedded in an epon analog resin. Ultrathin sections were cut using a Diatome diamond knife (Diatome, USA, Hatfield, PA) on a Leica Ultracut S ultramicrotome (Leica, Vienna, Austria). Sections were collected on copper grids and further contrasted with lead citrate and viewed on a JEM 1400 electron microscope (JEOL, USA, Inc., Peabody, MA) operated at 120 kV. Images were recorded with a Veleta 2K x2K digital camera (Olympus-SIS, Germany). nanoparticle-nanotubes were introduced to the media for 30 minutes at 1 mg/L, and then washed thoroughly and replaced with fresh media. After 6 hours, cells were washed with serum-free media then fixed with a modified Karmovsky's fix of 2.5% glutaraldehyde, 4% paraformaldehyde and 0.02% picric acid in 0.1M sodium cacodylate buffer at pH 7.2. Following a secondary fixation in 1% osmium tetroxide and 1.5% potassium ferricyanide, samples were dehydrated through a graded ethanol series, and embedded in an epon analog resin. Ultrathin sections were cut using a Diatome diamond knife (Diatome, USA, Hatfield, PA) on a Leica Ultracut S ultramicrotome (Leica, Vienna, Austria). Sections were collected on copper grids and further contrasted with lead citrate and viewed on a JEM 1400 electron microscope (JEOL, USA, Inc., Peabody, MA) operated at 120 kV. Images were recorded with a Veleta 2K x2K digital camera (Olympus-SIS, Germany). nanoparticle-nanotubes were introduced to the media for 30 minutes at 1 mg/L, and then washed thoroughly and replaced with fresh media. After 6 hours, cells were washed with serum-free media then fixed with a modified Karmovsky's fix of 2.5% glutaraldehyde, 4% Fluorescence microscopy of live cells Standard fluorescence imaging in the UV-visible emission range was performed on the hyperspectral microscope by using an XCite Series 120Q lamp as the light source and a QiClick CCD camera (QImaging) directly attached to a c-mount on a separate port of the microscope. Fluorescence filter sets from Chroma Technology and Semrock were used. Confocal imaging was performed on a Zeiss LSM 880, AxioObserver microscope equipped with a Plan- Apochromat 63× Oil 1.4 NA differential interference contrast (DIC) M27 objective in a humidified chamber at 37 °C. Z-stacks were obtained using a step size of 198-220 nm. Titrations of DNA-nanotube complexes with PEG-conjugated lipids Unsorted ss(GT)6-DNA-SWCNT samples were diluted to mg/L in PBS and incubated with various concentrations (0-5M) of two polyethylene glycol (PEG)-conjugated lipids (Cholesterol-PEG 600, “Cholesterol-PEG”, Sigma Aldrich; C16 PEG750 Ceramide, Avanti Lipids). Samples were incubated for 2 hours at 37 °C. Photoluminescence spectra were acquired with 2-second exposure times under 730 nm laser excitation. Polyethylene glycol, with molecular weights of 600 or 750 kDa, diluted in PBS, were used as controls to test for non- specific interactions. To test the effect of lowered pH on sensor performance, samples were diluted in a 100 mM pH 5.5 acetate buffer instead of PBS. Fluorescence spectroscopy of carbon nanotubes in solution Fluorescence emission spectra from aqueous solutions of SWCNTs were acquired using a home- built apparatus consisting of a tunable white light laser source, inverted microscope, and InGaAs NIR detector.54 The SuperK EXTREME supercontinuum white light laser source (NKT p p g ( Photonics) was used with a VARIA variable bandpass filter accessory capable of tuning the output 500 – 825 nm with a bandwidth of 20 nm. During the course of the measurements, the excitation wavelength remained at 730 nm, close to the resonant excitation maximum of the DNA-encapsulated (8,6) nanotube species. The light path was shaped and fed into the back of an inverted IX-71 microscope (Olympus) where it passed through a 20x NIR objective (Olympus) and illuminated a 100 µL nanotube sample at a concentration of 0.2 mg/L in a 96-well plate (Corning). With an exposure time of 1 second, the emission from the nanotube sample was collected through the 20x objective and passed through a dichroic mirror (875 nm cutoff, Semrock). The light was f/# matched to the spectrometer using several lenses and injected into an Isoplane spectrograph (Princeton Instruments) with a slit width of 410 µm which dispersed the emission using a 86 g/mm grating with 950 nm blaze wavelength. The spectral range was 930 – 1369 nm with a resolution of ~0.7 nm. The light was collected by a PIoNIR InGaAs 640 x 512 pixel array (Princeton Instruments). A HL-3-CAL-EXT halogen calibration light source (Ocean Optics) was used to correct for wavelength-dependent features in the emission intensity arising from the spectrometer, detector, and other optics. A Hg/Ne pencil style calibration lamp (Newport) was used to calibrate the spectrometer wavelength. Background subtraction was conducted using a well in a 96-well plate filled with DI H2O. Following acquisition, the data was processed with custom code written in Matlab which applied the aforementioned spectral corrections, background subtraction, and was used to fit the data with Lorentzian functions. Nanotube chirality and DNA sequence-dependent response to LDL Unsorted DNA-SWCNT samples were diluted to 2 mg/L in PBS and incubated with 0.5 mg/mL LDL (Alfa Aesar) for 18 hours at 37 °C. Chirality-separated samples were diluted to mg/L in PBS and incubated with 0.5 mg/mL LDL for 18 hours at 37 °C. Controls were incubated with no LDL present. Photoluminescence spectra were acquired with 2-second exposure times. Calculation of normalized Simpson’s Index The Simpson’s index is a diversity index used to measure the richness and evenness of a basic data type.62 The diversity index is maximized when all types of data are equally abundant. When applied to microbiology, the Simpson’s Index is referred to as the Hunter-Gaston index.62 In our application, 1 1 1 ( 1) ( 1) S j j j D n n N N     where D is the diversity index, N is the total number of pixels within each cell with detectable nanotube emission, S is the total number of histogram bins, and nj is the total number of pixels within the jth bin. We obtained the Simpson’s Index for each cell, SIj. For the set of SIj calculated for all the cells in an experiment, we normalized the value obtain the normalized Simpson’s Index, nSI. Cell culture reagents and conditions RAW 264.7 TIB-71 cells (ATCC, Manassas, VA) were grown under standard incubation conditions at 37˚C and 5% CO2 in sterile, filtered DMEM with 10% heat-inactivated FBS, 2.5% HEPES, 1% glutamine, and 1% penicillin/streptomycin (all from Gibco). For studies performed with homozygous mutant NPC, compound mutant heterozygote NPC, or wild-type fibroblasts, the respective cell lines, GM18453, GM03123 or GM05659 (Coriell, Camden, NJ) were cultured in MEM with 10% FBS, 2.5% HEPES, and 1% glutamine. Cells were plated on glass bottom petri dishes or lysine-covered glass dishes (MatTek) for fibroblasts. U2OS-SRA cells Chirality- separated SWCNTs were added at 0.2 mg/L in cell culture media (70 µL total volume) and incubated with cells for 30 minutes at 37 °C. This corresponds to approximately 0.5 picogram of SWCNT per cell, for a 50% cell confluency in a 9 mm diameter glass-bottom dish. The same procedure was used for unsorted SWCNTs with a concentration of 1 mg/L. These concentrations were chosen because they were experimentally observed to be the minimum concentration needed to obtain strong reporter signal from all of the cell lines used in the manuscript. An incubation time of 30 minutes was chosen because it was the minimal duration that resulted in a strong reporter signal from all the cell lines used in the manuscript. Cells were imaged immediately, or trypsinized (Gibco), and re-plated on a fresh petri dishes before hyperspectral imaging. All cells were used at 50-70% confluence. Filipin staining of NPC1 patient-derived fibroblasts Cells were fixed with 4% paraformaldehyde for 15 minutes, washed 3x with PBS, and stained with Filipin III (Sigma) at a concentration of 50 µg/mL for 20 minutes. The cells were then washed 3x with PBS and imaged using a DAPI filter cube. Cell viability and proliferation assays RAW 264.7 macrophages were seeded in untreated 96-well plates at 7,000 cells per well. The reporter was introduced to the cells at 0.2 mg/L. Reporter, vehicle (0.027 MSSC + 0.1 M NaSCN), or hydrogen peroxide-treated cells were incubated (times indicated), washed, and detached from the plate with Versene (1x PBS without Mg2+/Ca2+, 5mM EDTA, 2% FBS), pelleted, and incubated with Annexin V Alexa Fluor and Propidium Iodide (Life Technologies). Cells were analyzed by imaging cytometry (Tali) to quantify cell number and fluorophore content. For proliferation assays, RAW 264.7 macrophages treated with 0.2 mg/L ss(GT)6-(8,6)- SWCNT or vehicle were seeded at 150,000 cells on 100 mm diameter untreated culture dish on day zero. After settling for 10 hours, cells were harvested with Versene (1x PBS without Mg2+/Ca2+, 5mM EDTA, 2% FBS) and by mechanical tapping to remove cells from the surface, stained with Calcein AM, and counted for the initial seeding density. Media was replaced every 2 days. At each 24 hour period, cells were harvested as before and counted. Cell counts represent Calcein AM positive (live) cells. Lipidomics analyses Six T-175 flasks were seeded at <50% confluence with RAW 264.7 macrophages in their fifth passage. Three flasks each were treated as follows: control (untreated) or carbon nanotube treated (0.2 mg/L for 30 minutes, washing, and incubating for 6 hours). Cells were harvested (approx. 80% confluence in each set of flasks) by manual scraping. The flask contents corresponding to each condition were pooled into separate conical tubes and spun to pellet, washed in PBS containing protease inhibitor cocktail (Thermo-Pierce, 88666), pelleted, and flash frozen in a dry ice/IPA slurry with a small head volume of the PBS/inhibitor. For cell fractionation we adapted a sucrose/iodixanol equilibrium gradient centrifugation procedure for lysosome separation (Thermo, 89839). Briefly, flash frozen cells were thawed and suspended in 2x cpv of PBS/inhibitor, vortexed with reagent included in the kit, and Dounce homogenized with a cooled, tight fitting pestle using 90 strokes (starting cell material was >500 mg). After homogenization, reagent was added, tube inverted, and then centrifuged at 4˚C, 500x rcf for 10 minutes. The pellet was stored as the debris/nuclear fraction in all experiments. The supernatant was taken for subsequent ultracentrifugation. Briefly, one day before ultracentrifugation, a sucrose/iodixanol gradient (bottom to top: 30, 27, 23, 20, 17%) was layered into 12 mL polyallomer tubes (Thermo, 03699) and allowed to equilibrate in a cold room inside the metal buckets of an appropriate hanging-bucket rotor. The supernatant from above was mixed with the sucrose/iodixanol gradient to make a final sample density of 15% (total volume, 1 mL), which was then gently layered onto the top of the pre-formed gradient. The buckets were then sealed and moved into the ultracentrifuge using the following settings: ~135,000 rcf (32,000 rpm), 2.5 hours running time, acceleration/deceleration 9/5, 4˚C. The fractionated cell supernatant was removed from the top, and pipetted into six fractions based on volume removed from the ultracentrifuge tube. The volume removed from top to bottom was kept constant across the three conditions. Fractions were frozen until analysis. from the ultracentrifuge tube. The volume removed from top to bottom was kept constant across the three conditions. Fractions were frozen until analysis. Each of the six fractions, plus the nuclear/debris fraction (7 total/condition) was analyzed for the three conditions (21 fractions total). To quantify total protein, a standard curve was produced using BSA mixed into the sucrose/iodixanol gradient (Bradford assay background versus varying gradient was not different). Lipidomics analyses The difference was the measurement, which was compared to a standard curve that used Oleic acid (Sigma, O1008), prepared using the above protocol, as a model unsaturated hydrocarbon material. Total lipid levels were normalized by total protein content. added 200 μL ~98% sulfuric acid. The dried contents were coated with the acid by tipping the vial and vortexing. The vial was placed into a 100˚C mineral oil bath for 20 minutes. The resulting brown/black material was rapidly cooled in wet ice slurry for at least 5 minutes, and 100 μL was placed side-by-side into a 96-well glass plate. To one well, 50 μL of the phospho- vanillin reagent was added, mixed, and the plate incubated in the dark for 12 minutes. Absorption of each well (reagent reacted and sulfuric acid background) was taken at 535 nm. p g g The difference was the measurement, which was compared to a standard curve that used Oleic acid (Sigma, O1008), prepared using the above protocol, as a model unsaturated hydrocarbon material. Total lipid levels were normalized by total protein content. Lipidomics analyses 1x Bradford reagent at room temperature was mixed 1:1 with standard, allowed to incubate in the dark for 30 minutes, and the absorption measured at 595 nm. Each of the fractions was analyzed in this manner after addition of 0.2% IPEGAL CA- 630 (non-ionic detergent) to solubilize bound proteins. Cholesterol quantification was performed (Sigma, MAK043) using a coupled enzyme reaction between cholesterol oxidase and peroxidase with a proprietary colorimetric probe. Cholesterol esterase was used before the reactions to ensure total cholesterol was measured. Briefly, a 3-phase extraction was performed on each sample fraction (7:11:0.1 chloroform:isopropanol:IPEGAL CA-630). The top aqueous phase and interphase were removed, and the bottom organic phase was vacuum dried. The dried fractions were resuspended in provided buffer and reacted for 1 hour at 37˚C with the supplied reagents and the absorption read at 570 nm. This was compared to a standard curve. Cholesterol levels were normalized by total protein content as measured by the Bradford assay. Cholesterol quantification was performed (Sigma, MAK043) using a coupled enzyme reaction between cholesterol oxidase and peroxidase with a proprietary colorimetric probe. Cholesterol esterase was used before the reactions to ensure total cholesterol was measured. Briefly, a 3-phase extraction was performed on each sample fraction (7:11:0.1 chloroform:isopropanol:IPEGAL CA-630). The top aqueous phase and interphase were removed, and the bottom organic phase was vacuum dried. The dried fractions were resuspended Total lipid (total unsaturated hydrocarbon, including cholesterol) was measured after chloroform extracting the samples as above. Briefly, a phospho-vanillin color producing reagent was made by dissolving 5 mg/mL vanillin (Sigma, V1104) in 200 μL neat ethanol, and adding this to the appropriate volume of 17% phosphoric acid. This reagent was stored cool in the dark until needed. Dried sample fractions in glass vials were processed as follows: to each vial was added 200 μL ~98% sulfuric acid. The dried contents were coated with the acid by tipping the vial and vortexing. The vial was placed into a 100˚C mineral oil bath for 20 minutes. The resulting brown/black material was rapidly cooled in wet ice slurry for at least 5 minutes, and 100 μL was placed side-by-side into a 96-well glass plate. To one well, 50 μL of the phospho- vanillin reagent was added, mixed, and the plate incubated in the dark for 12 minutes. Absorption of each well (reagent reacted and sulfuric acid background) was taken at 535 nm. Extraction and differentiation of bone marrow-derived macrophages Bone marrow derived cells (BMDM) were prepared from 6 week-old C57/Bl6 mice and cultured in the presence of 10 ng/ml of recombinant colony stimulating factor-1 (CSF-1).64 Cells were collected 3, 5 and 7 days post-isolation and submitted to flow cytometry analysis for expression of the differentiation markers Gr-1 (monocytes/granulocytes-1/200), Cd11b (macrophages- 1/200), F4/80 (mature macrophages-1/50). Cells were incubated with 1 µl of Fc Block (BD Biosciences) for every million cells for at least 15 minutes at 4°C. Cells were then stained with the appropriate antibodies (BD Biosciences) for 20 minutes at 4°C, washed with FACS buffer, and resuspended in FACS buffer containing DAPI (5 mg/ml diluted 1:5,000) for live/dead cell exclusion.65 LysoTracker-nanotube colocalization RAW 264.7 or BMDM macrophages were incubated with Cy5-ss(GT)6-HiPco nanotubes for 30 minutes at a concentration 1 mg/L. The cells were then washed 3x with PBS and placed in fresh cell media. Six hours later, the cells were incubated with 5 nM LysoTracker Green DND-26 (Life Technologies) for 15 minutes in cell media, washed 3x with PBS, and imaged immediately in fresh PBS. The FITC or Cy5 channels were used for LysoTracker Green or Cy5-ss(GT)6- HiPco nanotubes, respectively. Plates of cells containing only Cy5-ss(GT)6-HiPco nanotubes or LysoTracker Green were used as controls to test for bleed-through across channels. Atomic force microscopy (AFM) A stock solution of ss(GT)6-(8,6)-SWCNTs at 7 mg/L in 100 mM NaCl was diluted 20x in dH2O and plated on a freshly cleaved mica substrate (SPI) for 4 minutes before washing with 10 mL of dH2O and blowing dry with argon gas. An Olympus AC240TS AFM probe (Asylum Research) in an Asylum Research MFP-3D-Bio instrument was used to image in AC mode. Data was captured at 2.93 nm/pixel XY resolution and 15.63 pm Z resolution. ASSOCIATED CONTENT The authors declare no competing financial interest. The authors declare no competing financial interest. Cell line source and authentication RAW 264.7 and U2OS cells were acquired from ATCC, and were tested for mycoplasma contamination by the source. Primary bone-marrow derived monocytes were tested for mycoplasma contamination using Dapi staining. Patient-derived fibroblasts were obtained from Coriell and tested for mycoplasma contamination by the source. Code availability Matlab code for the data analysis in this manuscript is available upon request, by contacting the corresponding author (hellerd@mskcc.org). Statistics Statistical analysis was performed with GraphPad Prism version 6.02. All data met the assumptions of the statistical tests performed (i.e. normality, equal variances, etc.). Experimental variance was found to be similar between groups using the F-test and Brown-Forsythe test for unpaired t-tests and one way ANOVAs, respectively. To account for the testing of multiple hypotheses, one way ANOVAs were performed with Dunnet’s, Tukey’s, or Sidak’s post tests when appropriate. Sample size decisions were based on the instrumental signal-to-noise ratios. Supporting Information The supporting information is available free of charge on the ACS Publications website and contains supplementary figures (S1-38), videos (1-3) and a supplementary table. AUTHOR INFORMATION Corresponding Author *Email: hellerd@mskcc.org ACKNOWLEDGEMENTS This work was supported by the NIH Director’s New Innovator Award (DP2-HD075698), NIH grants R01- HL093324, R01CA148967, R01CA181355, P20-GM103430 and P30 CA008748 cancer center support grant, the Rhode Island Foundation (20164347) the Anna Fuller Fund, the Louis V. Gerstner Jr. Young Investigator’s Fund, The Expect Miracles Foundation - Financial Services Against Cancer, the Experimental Therapeutics Center, Mr. William H. Goodwin, Mrs. Alice Goodwin, the Commonwealth Foundation for Cancer Research, the Honorable Tina Brozman Foundation, the Alan and Sandra Gerry Metastasis Research Initiative, Cycle for Survival, the Frank A. Howard Scholars Program, the Ara Parseghian Medical Research Foundation, and the Center for Molecular Imaging and Nanotechnology at Memorial Sloan Kettering Cancer Center. P.V.J. was supported by an NIH NCI-T32 fellowship (2T32CA062948- 21). D.R. was supported by an American Cancer Society 2013 Roaring Fork Valley Research Fellowship. T.V.G. was supported by the Frank Lappin Horsfall Jr. Fellowship. C.P.H. was supported in part by National Cancer Institute (NCI) Grant NIH T32 CA062948, J.M. was supported by the U.S. Department of Energy (DOE), Office of Science, Basic Energy Sciences (BES) under Award DE-SC0013979. L.A. was supported by the American Brain Tumor Association. J.B. was supported by a Tow Fellowship Award from the Center for Molecular Imaging and Nanotechnology, Memorial Sloan Kettering Cancer Center. DBI was supported by fellowship F31-DK104631 from the NIH. We thank the Molecular Cytology Core Facility at Memorial Sloan Kettering Cancer Center and the Electron Microscopy & Histology Core Facility at Weill Cornell Medical College. Use of the high-performance computing capabilities of the Extreme Science and Engineering Discovery Environment (XSEDE), supported by the National Science Foundation (NSF) grant numbers TG-MCB-120014 and TG-MCB-130013, is This work was supported by the NIH Director’s New Innovator Award (DP2-HD075698), NIH grants R01- HL093324, R01CA148967, R01CA181355, P20-GM103430 and P30 CA008748 cancer center support grant, the Rhode Island Foundation (20164347) the Anna Fuller Fund, the Louis V. Gerstner Jr. Young Investigator’s Fund, The Expect Miracles Foundation - Financial Services Against Cancer, the Experimental Therapeutics Center, Mr. William H. Goodwin, Mrs. Alice Goodwin, the Commonwealth Foundation for Cancer Research, the Honorable Tina Brozman Foundation, the Alan and Sandra Gerry Metastasis Research Initiative, Cycle for Survival, the Frank A. Howard Scholars Program, the Ara Parseghian Medical Research Foundation, and the Center for Molecular Imaging and Nanotechnology at Memorial Sloan Kettering Cancer Center. P.V.J. was supported by an NIH NCI-T32 fellowship (2T32CA062948- 21). D.R. gratefully acknowledged. We also thank Y. Shamay, A. Erez, R. Williams, R. Langenbacher, and J. Shah for helpful discussions, and J. Bartlett for aid in manuscript preparation. REFERENCES (1) Xu, H.; Ren, D. Lysosomal Physiology. Annu. Rev. Physiol. 2015, 77, 57–80. (2) Parenti, G.; Andria, G.; Ballabio, A. Lysosomal Storage Diseases: From Pathophysiology to Therapy. Annu. Rev. Med. 2015, 66, 471–86. py , (3) Grayson, M. Lysosomal Storage Disorders. Nature 2016, 537, S145–S145. (4) J W G L Ch l l d A h l i Cli Li id l 2 (3) Grayson, M. Lysosomal Storage Disorders. Nature 2016, 537, S145–S145. (4) Jerome, W. G. Lysosomes, Cholesterol and Atherosclerosis. Clin. Lipidol. 2010, 5, 853– 865. ( ) y , y g , , (4) Jerome, W. G. 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T.V.G. was supported by the Frank Lappin Horsfall Jr. Fellowship. C.P.H. was supported in part by National Cancer Institute (NCI) Grant NIH T32 CA062948, J.M. was supported by the U.S. Department of Energy (DOE), Office of Science, Basic Energy Sciences (BES) under Award DE-SC0013979. L.A. was supported by the American Brain Tumor Association. J.B. was supported by a Tow Fellowship Award from the Center for Molecular Imaging and Nanotechnology, Memorial Sloan Kettering Cancer Center. DBI was supported by fellowship F31-DK104631 from the NIH. We thank the Molecular Cytology Core Facility at Memorial Sloan Kettering Cancer Center and the Electron Microscopy & Histology Core gratefully acknowledged. We also thank Y. Shamay, A. Erez, R. Williams, R. Langenbacher, and J. Shah for helpful discussions, and J. Bartlett for aid in manuscript preparation. REFERENCES A Novel Alkyne Cholesterol to Trace Cellular Cholesterol Metabolism and Localization. J. Lipid. Res. 2014, 55, 583–591. 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https://openalex.org/W4383706223	https://zenodo.org/records/8129159/files/SIES%200802.pdf	English	null	THEORETICAL FOUNDATIONS OF NATIONS AND INTERNATIONAL RELATIONS	Zenodo (CERN European Organization for Nuclear Research)	2,023	cc-by	1,999	SCIENCE AND INNOVATION IN THE EDUCATION SYSTEM International scientific-online conference THEORETICAL FOUNDATIONS OF NATIONS AND INTERNATIONAL RELATIONS Kholikov Lazizjon Makhmud ugli Abstract: This article describes theoretical foundations for studying nations and international relations. Keywords: nation, interethnic relations, tolerance, close neighborhood, mutual cooperation, friendship, mentality, culture. About two thousand nations, peoples and tribes live in more than two hundred countries of the world. They communicate with each other in more than seven thousand languages, only more than four thousand of which have their own written language. Representatives of one nationality do not live in any country, representatives of dozens of nationalities live in one country. In particular, people of more than 140 nationalities live in Uzbekistan. At a new stage of development, the topic of relations between nations and nations becomes one of the most topical topics of socio-philosophical research. In the philosophical reflections of a number of philosophers, the recognition of interethnic relations, tolerance and good neighborly relations as one of the most important factors in ensuring sustainable development is of great importance. The nation occupies an important place in the system of socio-economic, national- cultural relations between people and is considered as part of social life. These relations not only provide social stability and harmony, regulate social relations, but also occupy an important place in the system of universal human values. “It is difficult to imagine mutual cooperation based on personal and social relations, strengthening the bonds of respect and friendship between different peoples and nations, without the achievements of language and modern science”1. As society develops, interethnic relations acquire a socio-philosophical content and develop. The continuous development of society creates a need for new socio-philosophical concepts in the system of interethnic relations. Such social improvement requires reliance on the customs, values and traditions of each ethnic group based on the law of succession. From this point of view, interethnic relations are both national and universal principles. Philosophers have given different definitions to the concepts of “nation” and “international relations”. So, let’s dwell on the following definitions, revealing the essence of these concepts. In this regard, the national encyclopedia 11 1 Ochilova B, Mamatkulov S. Ijtimoiy falsafa. –Toshkent: Noshir, 2016. –B. 108 of Uzbekistan defines the nation as follows: A nation is the highest peak of the ethnic history of the people, formed as a result of a long socio-economic, political and ethno-cultural process, within certain territorial limits. 2 Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimov A, Sultonov X, Jo‘rayev N, Vatan tuyg‘usi, -Toshkent: “O‘zbekiston”, 1996. – 443 b.; Jabborov I. O‘zbek xalqi etnografiyasi, -Toshkent: “O‘qituvchi”, 1994. – 320 b. 3 Adib Xolid O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob –Toshkent: Akademnashr, 2022. -B. 12 2 Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimov A, Sultonov X, Jo‘rayev N, Vatan tuyg‘usi, -Toshkent: “O‘zbekiston”, 1996. – 443 b.; Jabborov I. O‘zbek xalqi etnografiyasi, -Toshkent: “O‘qituvchi”, 1994. – 320 b. 3 Adib Xolid. O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob. –Toshkent: Akademnashr, 2022. -B. 19. 4 Otamurodov S. Globallashuv va millat. –Toshkent: “Yangi asr avlodi”, 2008. –B. 64. 5 Xonazarov. Q. Terrorchilar tenglik tarafdorimi?. // Tafakkur, -Toshkent: 2009, -№ 4 –B. 12. q , 3 Adib Xolid. O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob. –Toshkent: Akademn y , yg , , ; q g y , “O‘qituvchi”, 1994. – 320 b. 3 Adib Xolid. O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob. –Toshkent: Akademnashr, 2022. -B. SCIENCE AND INNOVATION IN THE EDUCATION SYSTEM International scientific-online conference THEORETICAL FOUNDATIONS OF NATIONS AND INTERNATIONAL RELATIONS , based on the unity of language and identity, a form of social unity that is formed on the basis of a certain culture, consciousness and mentality. Nation means the historical unity of people, economic life, unity of language, territory, harmony and commonality of culture, mind, spirit2. The philosophical essence of this definition is that a nation is a socio- spiritual environment created on the basis of the unity of people, covering several categories of the social population, formed on the basis of harmony in the social space, respect for traditions and values. In the Encyclopedic Dictionary of Philosophy, A nation is an ethnic unity of people who create their own material and spiritual wealth as an independent subject, connected by social ties, living in a certain territory on the basis of the unity of the spirit of the language, spirituality, awareness of national identity. The nation lives steadily in a certain territory, develops in the course of historical development, is formed on the basis of the unity of the spiritual and spiritual potential of people. It is defined as the product of humanity’s birth and development. In general, the recognition of a nation by a nation occurred even before the emergence of national self-consciousness among the members of the nation3. The philosopher Sadulla Otamurodov believes that interethnic relations are a set of activities between different nationalities living in the territories of countries in the economic, socio-political, cultural and educational spheres4. The current process of socio-economic integration into the world community, the sharp increase in terrorist threats, bloody conflicts in different regions of the world give rise to conflicts. Now in most countries of the world structures have been created to combat this destructive force, they have been given specific goals and tasks5. In such fragile and complex situations, time calls for the need to fight together against the dangers emanating from various ideological and social networks. 12 2 Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimov A, Sultonov X, Jo‘rayev N, Vatan tuyg‘usi, -Toshkent: “O‘zbekiston”, 1996. – 443 b.; Jabborov I. O‘zbek xalqi etnografiyasi, -Toshkent: “O‘qituvchi”, 1994. – 320 b. 3 Adib Xolid. O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob. –Toshkent: Akademnashr, 2022. -B. 19. 4 Otamurodov S. Globallashuv va millat. –Toshkent: “Yangi asr avlodi”, 2008. –B. 64. 5 Xonazarov. Q. Terrorchilar tenglik tarafdorimi?. // Tafakkur, -Toshkent: 2009, -№ 4 –B. 12. 2 Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimo SCIENCE AND INNOVATION IN THE EDUCATION SYSTEM International scientific-online conference THEORETICAL FOUNDATIONS OF NATIONS AND INTERNATIONAL RELATIONS 12 19. 4 Otamurodov S. Globallashuv va millat. –Toshkent: “Yangi asr avlodi”, 2008. –B. 64. 5 Xonazarov. Q. Terrorchilar tenglik tarafdorimi?. // Tafakkur, -Toshkent: 2009, -№ 4 –B. 12. 12 19. 4 Otamurodov S. Globallashuv va millat. –Toshkent: “Yangi asr avlodi”, 2008. –B. 64. 5 Xonazarov. Q. Terrorchilar tenglik tarafdorimi?. // Tafakkur, -Toshkent: 2009, -№ 4 –B. 12. SCIENCE AND INNOVATION IN THE EDUCATION SYSTEM International scientific-online conference References: 1. Ochilova B, Mamatkulov S. Ijtimoiy falsafa. –Toshkent: Noshir, 2016. –B. 108 2 Sh K O‘ b k l h kll h T hk “Sh ” International scientific-online conference 1. Ochilova B, Mamatkulov S. Ijtimoiy falsafa. –Toshkent: Noshir, 2016. –B. 108 1. Ochilova B, Mamatkulov S. Ijtimoiy falsafa. –Toshkent: Noshir, 2016. –B. 108 2. Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimov A, Sultonov X, Jo‘rayev N, Vatan tuyg‘usi, -Toshkent: “O‘zbekiston”, 1996. – 443 b.; Jabborov I. O‘zbek xalqi etnografiyasi, -Toshkent: “O‘qituvchi”, 1994. – 320 b. 2. Shoniyozov K. O‘zbek xalqining shakllanish jarayoni. –Toshkent: “Sharq”, 2001. – 464 b.; Ibrohimov A, Sultonov X, Jo‘rayev N, Vatan tuyg‘usi, -Toshkent: “O‘zbekiston”, 1996. – 443 b.; Jabborov I. O‘zbek xalqi etnografiyasi, -Toshkent: “O‘qituvchi”, 1994. – 320 b. 3. Adib Xolid. O’zbekiston tavalludi: Ilk SSSR davrida millat, imperiya va inqilob. –Toshkent: Akademnashr, 2022. -B. 19. 4. Otamurodov S. Globallashuv va millat. –Toshkent: “Yangi asr avlodi”, 2008. –B. 64. 5. Xonazarov. Q. Terrorchilar tenglik tarafdorimi?. // Tafakkur, -Toshkent: 2009, -№ 4 –B. 12. 6. 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https://openalex.org/W2019364304	https://publications.goettingen-research-online.de/bitstream/2/33772/2/journal.pgen.1004306.pdf	English	null	Proper Actin Ring Formation and Septum Constriction Requires Coordinated Regulation of SIN and MOR Pathways through the Germinal Centre Kinase MST-1	PLOS genetics	2,014	cc-by	10,482	Abstract This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This research project was financially supported through the DFG Heisenberg fellowship SE1054/6-1, the DFG grant SE1054/7-1 and by the Go¨ttingen Graduate School for Neurosciences and Molecular Biosciences (GGNB) (DFG grant GSC 226/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: Stephan.Seiler@biologie.uni-freiburg.de * E-mail: Stephan.Seiler@biologie.uni-freiburg.de Yvonne Heilig1, Anne Dettmann1, Rosa R. Mourin˜ o-Pe´rez2, Kerstin Schmitt3, Oliver Valerius3, Stephan Seiler1,4* 1 Institute for Biology II – Molecular Plant Physiology, Albert-Ludwigs University Freiburg, Freiburg, Germany, 2 Department of Microbiology, Center for Scientific Research and Higher Education of Ensenada (CICESE), Baja California, Mexico, 3 Institute for Microbiology and Genetics, University of Goettingen, Goettingen, Germany, 4 Freiburg Institute for Advanced Studies (FRIAS), Albert-Ludwigs University Freiburg, Freiburg, Germany Abstract Nuclear DBF2p-related (NDR) kinases constitute a functionally conserved protein family of eukaryotic regulators that control cell division and polarity. In fungi, they function as effector kinases of the morphogenesis (MOR) and septation initiation (SIN) networks and are activated by pathway-specific germinal centre (GC) kinases. We characterized a third GC kinase, MST- 1, that connects both kinase cascades. Genetic and biochemical interactions with SIN components and life cell imaging identify MST-1 as SIN-associated kinase that functions in parallel with the GC kinase SID-1 to activate the SIN-effector kinase DBF-2. SID-1 and MST-1 are both regulated by the upstream SIN kinase CDC-7, yet in an opposite manner. Aberrant cortical actomyosin rings are formed in Dmst-1, which resulted in mis-positioned septa and irregular spirals, indicating that MST-1- dependent regulation of the SIN is required for proper formation and constriction of the septal actomyosin ring. However, MST-1 also interacts with several components of the MOR network and modulates MOR activity at multiple levels. MST-1 functions as promiscuous enzyme and also activates the MOR effector kinase COT-1 through hydrophobic motif phosphorylation. In addition, MST-1 physically interacts with the MOR kinase POD-6, and dimerization of both proteins inactivates the GC kinase hetero-complex. These data specify an antagonistic relationship between the SIN and MOR during septum formation in the filamentous ascomycete model Neurospora crassa that is, at least in part, coordinated through the GC kinase MST-1. The similarity of the SIN and MOR pathways to the animal Hippo and Ndr pathways, respectively, suggests that intensive cross-communication between distinct NDR kinase modules may also be relevant for the homologous NDR kinases of higher eukaryotes. Citation: Heilig Y, Dettmann A, Mourin˜o-Pe´rez RR, Schmitt K, Valerius O, et al. (2014) Proper Actin Ring Formation and Septum Constriction Requires Coordinated Regulation of SIN and MOR Pathways through the Germinal Centre Kinase MST-1. PLoS Genet 10(4): e1004306. doi:10.1371/journal.pgen.1004306 Editor: Geraldine Butler, University College Dublin, Ireland Editor: Geraldine Butler, University College Dublin, Ireland Received December 20, 2013; Accepted February 25, 2014; Published April 24, 2014 Copyright: 2014 Heilig et al. This is an open-access article distributed under the terms of the Creative Commons unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Copyright: 2014 Heilig et al. April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org Introduction activates a Cdc14p-like phosphatase, which triggers mitotic exit [9]. Moreover, activated Dbf2p relocates to the future site of septum formation, where the kinase is required for the assembly and constriction of the cortical actomyosin ring (CAR; [10,11]). The second group of NDR kinases controls polarity as compo- nents of a homologous morphogenesis network (the fungal MOR network; homologous to the animal Ndr1/2 pathway) [5]. In fungi this pathway plays a critical role in the polar organization of the actin cytoskeleton at cell ends, which seems, at least in part, to be achieved through regulation of Rho-type GTPases [12–15]. In addition, the MOR regulates the expression of multiple genes with cell wall-related functions in unicellular yeasts, including genes required for efficient cell separation [16–18]. Coordination of cell growth and division is a fundamental subject in biology. Successful cytokinesis relies on the coordinated assembly and activation of an actomyosin-based contractile ring, which must be regulated in a spatially and temporally precise manner [1–3]. The underlying molecular pathways are highly complex and involve a large number of components forming elaborate interactive networks. Central components of these networks are the highly conserved nuclear Dbf2p-related (NDR) kinases, which are important for morphogenesis and proliferation in all eukaryotes analyzed to date [4–6]. These kinases represent two functional subgroups in fungi and animals. Dbf2p subfamily members are effector kinases of the fungal septation initiation network (SIN; homologous to the animal Hippo pathway; [7]), which consists of a cascade of three kinases that connects cell cycle progression with the initiation of cytokinesis [8]. Activation of the SIN by a spindle pole body (SPB)-associated small GTPase Multiple data indicate that the SIN and MOR function in concert in complex and mutual antagonistic manner to coordinate the remodeling of the actin cytoskeleton during cell growth and division [5,10,11]. Inhibition of the fission yeast MOR network by the SIN has been shown to promote cytokinesis, while polar April 2014 \| Volume 10 \| Issue 4 \| e1004306 1 PLOS Genetics \| www.plosgenetics.org MST-1 Coordinates SIN and MOR Networks phenotypic characterization of mutants in these SIN components revealed that Dsid-1 and Dcdc-14 deletion strains behave differently than mutants in the remaining SIN components, suggesting that the presentation of the SIN as a linear kinase cascade may represent a simplified view. In this study, we characterized MST-1, the third germinal centre (GC) kinase present in N. The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components Germinal centre (GC) kinases represent a large family of functionally diverse proteins that can be grouped into eight subfamilies [44]. Fungal GC kinases represent three functionally distinct subgroups of subfamily III (Figure 1 A; Figure S1). N. crassa POD-6 and the related budding and fission yeast kinases Kic1p and Nak1, respectively, clustered together, in line with their conserved function as upstream components of the MOR network [38,45,46]. The second subgroup is represented by the SIN kinases Sid1/SID-1 [43,47]. Proteins of the third subgroup are most closely related to animal group III GC kinases. The fission yeast member Ppk11 was recently characterized as auxiliary factor of the MOR pathway that supports cell separation [48], while the N. crassa protein NCU00772/MST-1 had been implicated as part of the SIN in a preliminary analysis [49]. growth in interphase requires inhibition of the SIN by the MOR [19–21]. Moreover, the activity of the MOR is SIN-dependent [22,23], consistent with the hypothesis of the MOR acting as SIN effector after septum constriction to allow cell separation. Significantly, the animal Hippo and Ndr1/2 kinase pathways also have opposing effects on cell morphogenesis and proliferation [24– 26], indicating that this interdependence of the two NDR kinase modules is conserved among eukaryotes. In order to address these discrepancies, we compared localiza- tion pattern and mutant characteristics of MST-1/Dmst-1 with those of SIN and MOR components. Dmst-1 formed multiple, closely spaced septa similar to the hyperseptation defects observed in MOR mutants and produced abnormal cross walls in the form of cortical spirals in older hyphal segments (Figure 1 B). However, a MST-1-GFP fusion construct, which complemented all Dmst-1 defects, associated with spindle pole bodies (SPBs) and constricting septa (Figure 1 C; Video S1), a localization pattern characteristic for fungal SIN components. Moreover, the association of MST-1 with both SPBs was constitutive and cell cycle independent (Figure 1 D), consistent with the localization of the three N. crassa SIN kinases [43]. In contrast, a POD-6-GFP fusion protein localized at the hyphal tip in a dot-like structure in the Spitzenko¨rper, as membrane-associated apical crescent and around the septal pore (Figure 1 C; Figure S2), reflecting the localization pattern of other MOR components [41,42]. Neither POD-6 nor its target kinase COT-1 and regulatory subunit MOB-2A were detected at SPBs. Therefore, we compared the kinetics of cortex recruitment of the two NDR kinases with that of MST-1 during septum initiation (Figure 2). Author Summary Cytokinesis is a fundamental cellular process essential for cell proliferation of uni- and multicellular organisms. The molecular pathways that regulate cytokinesis are highly complex and involve a large number of components that form elaborate interactive networks. The fungal septation initiation network (SIN) functions as tripartite kinase cascade that connects cell cycle progression with the control of cell division. Mis-regulation of the homologous Hippo pathway in animals results in excessive proliferation and formation of tumors, underscoring the conservation and importance of these kinase networks. A second morphogenesis (MOR) pathway involves homologous components and is controlling cell polarity in fungi and higher eukaryotes. Here we show that the promiscuous functioning Ste20-related kinase MST-1 has a dual role in regulating SIN and MOR network function. Moreover, SIN and MOR coordination through MST-1 can be achieved in an enzyme-independent manner through hetero-dimer- ization of germinal centre kinases, providing an additional level for activity regulation of signaling networks that is not dependent on phosphate transfer. Given the func- tional conservation of NDR kinase signaling modules and their regulation, our work may define general mechanisms by which NDR kinase pathway are coordinated in fungi and higher eukaryotes. Introduction crassa and other fungi, which we show is important for proper CAR formation and septum constriction. We provide evidence that MST-1 functions in parallel to SID-1 as part of the SIN, but also that MST-1 has multiple functions in regulating the MOR. Consequently, MST-1 plays an important role in coordinating the antagonistic functions of the SIN and MOR pathways during septum formation. Author Summary Results The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components April 2014 \| Volume 10 \| Issue 4 \| e1004306 The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components For comparison, pod-6-defective cells produce multiple, closely spaced septa, while Dsid-1 cells are aseptate. Cell wall and septa were labeled with Calcofluor White. (C) Functional GFP fusion proteins of the three GC kinases SID-1, MST-1 and POD-6 localize as constricting rings at septa. SID-1 and MST-1, but not POD-6 also localize to spindle pole bodies (arrows), while POD-6 accumulated as dot in the distal region of the Spitzenko¨rper and as apex-associated crescent (Figure S2). Nuclei were labeled with histone H1-RFP and the cell wall was stained with Calcofluor White. (D) MST-1-GFP associated with SPBs of interphase nuclei as well as during early and late mitotic stages (as indicated by nuclear morphology). Nuclei were labeled with histone H1-RFP. doi:10.1371/journal.pgen.1004306.g001 crassa [35]. A functional BNI-1-GFP fusion construct formed cortical rings with uniform signal intensity in wt cells, and septum constriction was concentric, resulting in centrally positioned septal pores (Figure 3 A upper panel; Video S2). In contrast, BNI-1 frequently (i.e. in 38 of 47 septation events analyzed) formed asymmetric rings that led to acentric constriction and asymmetric septal pores in Dmst-1 (Figure 3 A lower panel; Videos S3, S4). BNI-1 also associated with extensive cortical Calcofluor White- labeled spirals (Figure 3 B), indicating that the secretory machinery is still correctly targeted and that growth of the cell wall at these sites is not inhibited through the mis-organized formin rings and spirals. Moreover, CAR assembly and constriction was directly monitored using a Lifeact-GFP construct recently developed for N. crassa [52]. Lifeact-GFP labeled a mesh of F-actin cables and patches named the septal actin tangle (SAT), which appeared approximately .5 min prior to septum constriction at the future septation site in wt (Figure 2; [52,53]). After 2–3 min, the SAT coalesced to form the CAR (Figure 3 C upper panel; Video S5). In contrast to wt, the F-actin meshwork was mis-organized and the in ca. 50% of round (yet fully viable) ascospores, in contrast to the typical pea-shaped progeny generated in wt6wt crosses (Figure S3). Moreover, Dmst-16Dmst-1 crosses resulted in the formation of empty perithecia lacking asci, and the formation of ascospores was abolished. We also observed synthetic behavior of Dmst-1 with SIN-, but not MOR-defective strains during sexual development. Dmst-16Dcot-1 crosses resulted in the expected segregation of round and normally shaped ascospores, although the total number of generated ascospores was reduced. In contrast, Dmst-16Ddbf-2 crosses generated empty perithecia and no ascospores. The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components Mutant 6wt crosses resulted April 2014 \| Volume 10 \| Issue 4 \| e1004306 2 PLOS Genetics \| www.plosgenetics.org MST-1 Coordinates SIN and MOR Networks Figure 1. MST-1 displays features reminiscent of SIN as well as MOR components. (A) Phylogenetic comparison of fungal GC kinases. The tree was generated by the neighbour-joining method based on a ClustalW alignment of the indicated S. cerevisiae, S. pombe and N. crassa proteins. HsMst3 and DmGck3 were used as examples for animal group III GC kinases. The GC kinase II DmHippo was used as outgroup member (multiple alignment parameters: open gap penalty 10.0, extend gap penalty 0.0, delay divergent 40%, gap distance 8, similarity matrix blosum). An extended phylogram of fungal GC kinases is found in Figure S1. (B) Phenotypic characteristics of Dmst-1, pod-6(ts) and Dsid-1 during septum formation. Note the presence of closely spaced septa (second panel) and abnormal spirals (third panel) in Dmst-1. For comparison, pod-6-defective cells produce multiple, closely spaced septa, while Dsid-1 cells are aseptate. Cell wall and septa were labeled with Calcofluor White. (C) Functional GFP fusion proteins of the three GC kinases SID-1, MST-1 and POD-6 localize as constricting rings at septa. SID-1 and MST-1, but not POD-6 also localize to spindle pole bodies (arrows), while POD-6 accumulated as dot in the distal region of the Spitzenko¨rper and as apex-associated crescent (Figure S2). Nuclei were labeled with histone H1-RFP and the cell wall was stained with Calcofluor White. (D) MST-1-GFP associated with SPBs of interphase nuclei as well as during early and late mitotic stages (as indicated by nuclear morphology). Nuclei were labeled with histone H1-RFP. doi:10.1371/journal.pgen.1004306.g001 Figure 1. MST-1 displays features reminiscent of SIN as well as MOR components. (A) Phylogenetic comparison of fungal GC kinases. The tree was generated by the neighbour-joining method based on a ClustalW alignment of the indicated S. cerevisiae, S. pombe and N. crassa proteins. HsMst3 and DmGck3 were used as examples for animal group III GC kinases. The GC kinase II DmHippo was used as outgroup member (multiple alignment parameters: open gap penalty 10.0, extend gap penalty 0.0, delay divergent 40%, gap distance 8, similarity matrix blosum). An extended phylogram of fungal GC kinases is found in Figure S1. (B) Phenotypic characteristics of Dmst-1, pod-6(ts) and Dsid-1 during septum formation. Note the presence of closely spaced septa (second panel) and abnormal spirals (third panel) in Dmst-1. The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components DBF-2 and MST-1 formed cortex-associated rings 2:5060:45 min (n = 17) and 2:5561:25 min (n = 19), respectively, prior to the start of septum constriction, which was monitored by plasma membrane invagination and defined as time point 0:00 min (labeled with FM4-64). In contrast, COT-1 was only observed after CAR constriction had started (0:3960:13 min; n = 15). Furthermore, the fusion protein strongly labeled the mature septal pore [41,42]. In contrast to the total separation of mother and daughter cells, which is characteristic for unicellular yeasts, the vast majority of fungi form mycelial colonies that consist of networks of branched hyphae and are compartmentalized by incomplete septa. The septal pores enable communication and transport of cytoplasm and organelles between adjacent cells. Despite the importance of septation for growth, proliferation and differentiation of molds, our understanding of septum formation and its regulation in filamentous fungi is highly fragmentary [27–29]. Genetic and biochemical analysis in the model molds Neurospora crassa and Aspergillus nidulans suggest that cell cycle-dependent signals of a subset of competent mitotic nuclei activate the SIN [30–32]. This, in turn, is critical for the cortical localization of a RHO-4/BUD-3 GTPase module complexed by its anillin-type scaffold BUD-4, which allows the generation of the CAR through a second RHO- 4/RGF-3 GTPase complex [33–35]. In addition to these positive regulators of septum formation, several negative regulators were identified in N. crassa, whose mutation result in hyperseptated strains [36]. Most notably are LRG-1, a RHO-1-specific GTPase activating protein [14] and COT-1, POD-6 and two MOB-2 proteins, the central elements of the N. crassa MOR network [37– 39]. Moreover, components of the RHO-1 module and the MOR network localize to forming septa, providing additional support for a function of these proteins during septation [14,40–42]. We have recently determined that a hierarchical, tripartite SIN cascade, consisting of CDC-7, SID-1 and DBF-2, together with their regulatory subunits CDC-14 and MOB-1, respectively, are the central components of the SIN and are essential for septum formation in N. crassa [39,43]. However, the comparative In addition to the observed septation defects in subapical hyphal regions, Dmst-1 displayed only minor vegetative abnormalities. Mycelial extension rates, colony behavior and conidiation pattern were similar to wild type (wt; data not shown). However, sexual differentiation was affected in Dmst-1. The germinal centre kinase MST-1 displays features reminiscent of SIN as well as MOR components The same synthetic effect was observed in crosses of Dmst-1 with Dsid-1 or Dcdc-7, while crosses of Dmst-16Dpod-6 produced round as well as pea-shaped spores. MST-1 controls proper CAR formation Fungal septum formation is driven through the constriction of a cortex-associated actomyosin ring (CAR; [50,51]). Thus, we compared the dynamics of septum formation in wt and Dmst-1 by monitoring the behavior of the sole formin BNI-1 present in N. April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 3 MST-1 Coordinates SIN and MOR Networks Figure 2. Kinetics of cortex association of SIN and MOR components DBF-2, MST-1 and COT-1 during septum forma- tion. (A) Composite images of time series of the indicated proteins during septum formation. The start of plasma membrane invagination was monitored with FM4-64 and defined as time point 0:00 min. Arrows indicate the time point when the GFP-fusion proteins occurred first at the cell cortex and arrowhead the time point of complete CAR constriction. (B) Schematic representation of the chronology of emergence of the SIN and MOR at the cell cortex relative to the actin cytoskeleton. doi:10.1371/journal.pgen.1004306.g002 while only CDC-7 co-purified with MST-1. Next, we determined physical interactions of the SIN components in yeast two hybrid (Y2H) assays (Figure 4 B). DBF-2 and SID-1 directly interacted with their regulatory subunits MOB-1 and CDC-14, respectively, and both kinases were able to form homo-dimers. In addition, a weak interaction was detected between the two kinases. Interest- ingly, the upstream kinase CDC-7 did not directly interact with SID-1, but was connected to the GC kinase via its adaptor CDC- 14. Finally, we detected a physical interaction of MST-1 and CDC-7 in Y2H tests (Figure 4 B), which was confirmed by co- immunoprecipitation (co-IP) experiments (Figure 4 C). p p ( ) p ( g ) Our GFP-trap + MS data indicated that SID-1 and MST-1 only co-purified together, when we used CDC-7 as bait, while the two GC kinases behaved mutually exclusive in the other co-purifica- tions. Moreover, we were unable to co-purify SPG-1, CDC-14 and the DBF-2/MOB-1 complex with MST-1, suggesting the presence of distinct CDC-7/SID-1 and CDC-7/MST-1 complexes. In order to further strengthen this assumption, we performed in vitro displacement assays, in which a GFP-trapped CDC-7-GFP/HA- GC kinase complex precipitated from a strain co-expressing both tagged kinases was incubated with excessive amounts of separately purified second GC kinase (Figure 4 D, E). The composition of the resulting CDC-7 complex was determined by Western blot analysis after pelleting the sepharose bead-associated proteins. MST-1 regulates the MOR network by two distinct mechanisms Based on the presented genetic, biochemical and life imaging analyses, we conclude that MST-1 is part of the SIN and functions in parallel to SID-1. However, the hyperseptation defects observed in Dmst-1 partly phenocopied characteristics of MOR mutants. Consistent with the hypothesis that MST-1 may regulate both NDR kinase pathways was the ability of purified MST-1 to activate COT-1 in addition to its activation of DBF-2 (Figure 6 A, B). As control, we confirmed that SID-1 only stimulated DBF-2, while POD-6 was specific for COT-1, confirming that MST-1 has a specific function as promiscuous activator of both pathways. Moreover, MST-1 was unable to stimulate DBF-2 and COT-1 variants, in which their hydrophobic motif phosphorylation sites were modified, indicating that MST-1 activates both target kinases by hydrophobic motif phosphorylation. MST-1 controls proper CAR formation In these assays, we were able to replace CDC-7-associated SID-1 with MST-1 and vice versa, indicating that the two GC kinases form mutually exclusive assemblies with CDC-7. Based on these interaction data, we hypothesized that the two GC kinases function in parallel as part of the SIN. Consistent with this prediction, we determined that purified SID-1 and MST-1 both stimulated DBF-2 activity in vitro (Figure 5 A). As predicted for the stepwise function of the SIN kinase cascade, addition of CDC-7 to the SID-1/DBF-2 kinase mixture further stimulated DBF-2, while addition of kinase-dead CDC-7(D195A) did not. In contrast, CDC-7 inhibited MST-1-driven stimulation of DBF-2, and this inhibition did not require functional CDC-7. As control, we confirmed that the kinase activity of CDC-7 was essential for complementation of the deletion defects of Dcdc-7 (data not shown). Strikingly, we found that septum association of CDC-7, but not its localization at SPBs was dependent on the functionality of CDC-7 (Figure 5 B). In contrast, the localization of DBF-2 and MST-1 at SPBs and septa did not require enzyme activity (Figure S4), potentially because homo-dimerization of the kinase-dead version with a functional kinase variant allowed correct localiza- tion in a wt background. Figure 2. Kinetics of cortex association of SIN and MOR components DBF-2, MST-1 and COT-1 during septum forma- tion. (A) Composite images of time series of the indicated proteins during septum formation. The start of plasma membrane invagination was monitored with FM4-64 and defined as time point 0:00 min. Arrows indicate the time point when the GFP-fusion proteins occurred first at the cell cortex and arrowhead the time point of complete CAR constriction. (B) Schematic representation of the chronology of emergence of the SIN and MOR at the cell cortex relative to the actin cytoskeleton. doi:10.1371/journal.pgen.1004306.g002 actin cables were irregularly distributed in Dmst-1, and the SAT to CAR transition lasted much longer (8:3062:00 min in Dmst-1 (n = 12) compared to 2:3060:30 min in wt (n = 15)). This failure of correct CAR assembly resulted in acentric constriction and asymmetric position of septal pores or the formation of open actin spirals, which were unable to constrict (Figure 3 C lower panel, Figure 3 D; Videos S6, S7). MST-1 regulates the MOR network by two distinct mechanisms April 2014 \| Volume 10 \| Issue 4 \| e1004306 The SIN kinase CDC-7 regulates SID-1 and MST-1 in an antagonistic manner The SIN kinase CDC-7 regulates SID-1 and MST-1 in an antagonistic manner g GFP-trap affinity purification experiments coupled to mass spectrometry (MS) allowed determining MST-1-interacting pro- teins and its relationship to the SIN (Figure 4 A; Table S1). Precipitates of a DBF-2-GFP fusion recovered all components of the central SIN cascade, including its regulatory subunit MOB-1, the two upstream kinases CDC-7 and SID-1 and its adaptor CDC- 14, and the predicted GTPase SPG-1/NCU08878. In contrast, SID-1-GFP recovered only its subunit CDC-14 and the upstream components CDC-7 and SPG-1, but not the DBF-2/MOB-1 complex, indicating a less robust interaction with the SIN effector kinase. MST-1 did not co-purify these two kinases. However, CDC-7-GFP co-precipitated both GC kinases SID-1 and MST-1 in addition to the GTPase SPG-1 and the SID-1 adapter CDC-14, Because we had detected several POD-6-specific peptides in one of the two MST-1-GFP GFP-trap+MS analyses (Table S1), we April 2014 \| Volume 10 \| Issue 4 \| e1004306 April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 4 Figure 3. MST-1 is required for proper contractile actin ring formation. (A) 4D reconstruction of z-stacks in time lapse series revealed cortical, concentrically constricting BNI-1-GFP rings in wt cells, which resulted in centrally positioned septal pores. In contrast, BNI-1 formed asymmetric and frequently open BNI-1-GFP rings in Dmst-1 that led to acentric CAR constriction and asymmetric septal pores. (B) 3D reconstruction of z-stacks illustrates BNI-1-GFP association with extensive cortical Calcofluor White-labeled spirals in Dmst-1. (C) Comparison of actin dynamics during CAR assembly and constriction in wt and Dmst-1. Lifeact-GFP labeled a dynamic meshwork of actin cables and patches around the future septation site in wt cells, which subsequently coalesced to form the CAR. The actin meshwork was mis-organized and irregularly distributed in Dmst-1 (D) 4D reconstruction of z-stacks in time lapse series visualized open actin spirals labeled by Lifeact-GFP, which were unable to constrict. Cell wall, septa and cortical spirals were labeled by Calcofluor White. doi:10.1371/journal.pgen.1004306.g003 MST-1 Coordinates SIN and MOR Networks MST-1 Coordinates SIN and MOR Networks Figure 3. MST-1 is required for proper contractile actin ring formation. (A) 4D reconstruction of z-stacks in time lapse series revealed cortical, concentrically constricting BNI-1-GFP rings in wt cells, which resulted in centrally positioned septal pores. In contrast, BNI-1 formed asymmetric and frequently open BNI-1-GFP rings in Dmst-1 that led to acentric CAR constriction and asymmetric septal pores. PLOS Genetics \| www.plosgenetics.org Discussion In these unicellular models, only the terminal kinase constitutively associates with both SPBs and relocates after its activation at the SPB to the septum, while the upstream kinases associate only with one SPB in a cell cycle-dependent manner and do not relocate [10,55]. This re-localization of the activated Ndr kinase is thought to be important for controlling the coordination between cell cycle progression and cell division. Strikingly, SIN activation in A. nidulans does not require its previous interaction with the SPB [32], and localization of the entire SIN at SPBs is cell cycle independent in N. crassa. Moreover, we found that the activity of CDC-7 is essential for its localization to septa, but not its association with the SPB. These data strongly suggest differential regulation of the SIN in uninuclear and syncytial fungi. The mechanism for positioning new septa in a multi-nuclear syncytium is currently unresolved [27,28]. Although the SIN is a prime candidate for transmitting nuclear signals that are required for the selection of future septation sites [27,28,30], we have currently no information if SPB association of the SIN has any function during vegetative hyphal growth. g g yp g MST-1 also interacts with and regulates several components of the MOR network. First, MST-1 activates the MOR effector kinase COT1 through hydrophobic motif phosphorylation, a mechanism previously shown for other fungal and animal NDR kinases [41,43,56–58]. This dual function of SIN and MOR regulation reflects NDR kinase signaling in higher eukaryotes, where the GC kinase Hippo has been shown to activate two NDR kinases that regulate cell proliferation as well as polar morpho- genesis, respectively [59–61]. Moreover, MST-1 can form heterodimers with the MOR kinase POD-6, and this interaction results in inhibition of the GC kinase complex. Thus, MST-1 can stimulate as well as inhibit MOR signaling. Although the functional significance of this dual function of MST-1 is yet unresolved, the N. crassa MOR likely has multiple functions during septation. First, MOR mutants are characterized by abundant, closely spaced septa, arguing for an early function of the MOR in the inhibition of septum initiation. However, COT1, its regulatory subunit MOB-2A, and POD-6 associate with the CAR only after the start of septum constriction and primarily label the mature septal pore (this study; [41,42]). Thus, the MOR may also be critical to regulate termination of constriction and proper pore formation/maintenance. The SIN kinase CDC-7 regulates SID-1 and MST-1 in an antagonistic manner (B) 3D reconstruction of z-stacks illustrates BNI-1-GFP association with extensive cortical Calcofluor White-labeled spirals in Dmst-1. (C) Comparison of actin dynamics during CAR assembly and constriction in wt and Dmst-1. Lifeact-GFP labeled a dynamic meshwork of actin cables and patches around the future septation site in wt cells, which subsequently coalesced to form the CAR. The actin meshwork was mis-organized and irregularly distributed in Dmst-1 (D) 4D reconstruction of z-stacks in time lapse series visualized open actin spirals labeled by Lifeact-GFP, which were unable to constrict. Cell wall, septa and cortical spirals were labeled by Calcofluor White. doi:10.1371/journal.pgen.1004306.g003 asked if MST-1 also interacted with the MOR pathway. Y2H experiments confirmed the physical interaction of MST-1 with POD-6 as well as the capability of both kinases to form homo- dimers through their kinase domains (Figure 7 A). Moreover, co- IP experiments suggested that the MST-1/POD-6 complex was more stable and withstood to more stringent wash conditions than the COT-1/POD-6 interaction (Figure 7 B). Moreover, POD-6 was replaced by MST-1 by adding separately purified MST-1 to the COT-1/POD-6 complex (Figure 7 C). Thus, a second mechanism of MST-1-dependent regulation of the MOR could involve dimerization of POD-6 and MST-1 and thereby inactivation of the GC kinase hetero-dimer. Indeed, when we titrated purified POD-6 with increasing levels of a kinase-dead version of MST-1(D157A) the activity of POD-6 decreased, while addition of a mock precipitate did not interfere with POD-6 activity (Figure 7 D). April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 5 MST-1 Coordinates SIN and MOR Networks PLOS Genetics \| www.plosgenetics.org 6 April 2014 \| Volume 10 \| Issue 4 \| e100430 PLOS Genetics \| www.plosgenetics.org 6 April 2014 \| Volume 10 \| Issue 4 \| e1004306 April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 6 MST-1 Coordinates SIN and MOR Networks Figure 4. CDC-7 forms mutually exclusive complexes with SID-1 and MST-1. (A) AP-MS data from two independent biological replicates were used for identification of predicted SIN components. * denotes % protein coverage and number of unique peptides of the identified protein. Only SIN components identified in both replicate purifications and absent from a control data set obtained in purifications with GFP-expressing cells are displayed (Table S1). GFP-fusion constructs used as bait proteins are in bold type. The SIN kinase CDC-7 regulates SID-1 and MST-1 in an antagonistic manner (B) Yeast two-hybrid tests with the indicated SIN constructs and summary schema of the observed interactions. (C) Reciprocal co-immunoprecipitation experiments of CDC-7-GFP and HA-MST-1 from cell extracts co-expressing both functionally tagged proteins indicate interaction of the two kinases. (D, E) GC-kinase displacement assays of precipitated CDC-7- GFP/GC-kinase complexes. Addition of a second, separately purified GC kinase, but not a mock-IP precipitate to these complexes reduced the abundance of the co-purified GC kinase. The upper panel displays a representative experiment, while three independent experiments are quantified in the lower graph. d i 10 1371/j l 1004306 004 doi:10.1371/journal.pgen.1004306.g004 doi:10.1371/journal.pgen.1004306.g004 septum initiation; i.e. the trigger required for septum placement/ initiation may not be terminated, but can signal into the neighboring area to allow the sequential formation of septa and/ or spirals if the SAT does not properly coalesce into a functional CAR (Video S4). Discussion septum initiation; i.e. the trigger required for septum placement/ initiation may not be terminated, but can signal into the neighboring area to allow the sequential formation of septa and/ or spirals if the SAT does not properly coalesce into a functional CAR (Video S4). GC kinases function as central components of the SIN and MOR networks, which have antagonistic functions in the regulation of septation in unicellular as well as filamentous fungi [5,8,20]. In this study, we report the characterization of MST-1, the third type III GC kinase present in fungi, and dissect its dual function during SIN and MOR signaling (summarized in Figure 8). Our results demonstrate that MST-1 is a SIN-associated kinase that physically interacts with CDC-7 and functions in parallel to SID-1 to activate DBF-2. However, the interaction data strongly suggest that MST-1 is not part of the central SIN cascade, but forms a distinct complex with CDC-7. As predicted for the tripartite kinase cascade, active CDC-7 is required to stimulate hydrophobic motif phosphorylation of DBF-2 by SID-1. In contrast, MST-1 activity is negatively regulated through CDC-7 in a mechanism that does not require active kinase, indicating that the regulation of SID-1 and MST-1 through CDC-7 is based on distinct mechanisms. A possible function of MST-1 in fine-tuning the SIN may be achieved in the generation of an incoherent type 4 feedforward loop through the two, parallel functioning, but negatively regulated CDC-7/SID-1 and CDC-7/MST-1 com- plexes that together activate DBF-2. This rarely found regulatory motif may allow for adaptive and thus robust SIN activity [54]. This hypothesis is supported by the phenotypic characteristics of N. crassa SIN mutants, which revealed less severe defects of sid-1 mutants compared to strains deficient for cdc-7 or dbf-2 [43]. In summary, these results indicate that the composition and function of the SIN as a linear kinase cascade represents a simplified view. Our biochemical data also imply that the CDC-7/MST-1 complex may function independently of the GTPase SPG-1, a speculation that requires further exploration. ( ) All SIN kinases, including MST-1, also localize in a cell cycle- independent manner to both SPBs. This pattern differs from the localization of homologous kinases in budding and fission yeast. Discussion Thus, although originally thought to constitute distinct modules, antagonistic cross-talk between the SIN and MOR occurs at multiple levels (this study; [19,21]). Further analysis is required to dissect their individual impact for the coordinated action of both networks during cell division. The maturation of the CAR in N. crassa is driven by the coalescence of a filamentous actin meshwork, called the SAT [50,52,53]. The association of all SIN components except CDC-7 approximately 3 min prior to the start of CAR constriction coincides with the time window of SAT to CAR transition, and thus the SIN is likely involved in this process. However, the aseptate nature of sin mutants precludes a detailed analysis of CAR function in these strains. Here we show that the SIN-associated kinase MST-1 plays a significant role in the proper assembly and constriction of the CAR. The defects generated by mst-1 mutants allow visualizing defects during the transition of SAT to CAR and imply a central function of the SIN in CAR maturation and in CAR constriction. Life imaging of actin as well as formin dynamics revealed two striking defects in Dmst-1. First, the CAR was frequently generated in an asymmetric manner, resulting in its acentric constriction and the formation of mispositioned septal pores. Moreover, the transition from the SAT to the CAR at the future septation site was delayed about 3-4-fold compared to wt cells, and the SAT frequently failed to coalesce into a functional CAR, generating constriction-defective spirals. In summary, we speculate that this complex phenotype is the result of mis-regulated April 2014 \| Volume 10 \| Issue 4 \| e1004306 7 PLOS Genetics \| www.plosgenetics.org MST-1 Coordinates SIN and MOR Networks Figure 5. The regulation of MST-1 and SID-1 by CDC-7 is based on distinct mechanisms. (A) In vitro DBF-2 activity assays after addition of the indicated kinases. MST-1-stimulated DBF-2 activity is inhibited by addition of CDC-7 or CDC-7(D195A), while SID-1-stimulation requires active CDC-7. Western blot analysis of the precipitated proteins was used to determine comparable kinase levels (n = 4). (B) A GFP fusion construct of wt CDC-7 localized to spindle pole bodies and accumulated around the mature septal pore, while a CDC7(D195A)-GFP construct exclusively localized to SPBs. Nuclei were labeled with histone H1-RFP and plasma membrane with FM4-64. doi:10.1371/journal.pgen.1004306.g005 Figure 5. The regulation of MST-1 and SID-1 by CDC-7 is based on distinct mechanisms. Materials and Methods The Matchmaker Two-Hybrid system 3 (Clontech, USA) was used according to the manufacturer’s instructions. cDNA of genes of interest for two hybrid tests was amplified with primers listed in Table S3 spanning the coding region from start to stop codon as annotated by the N. crassa database (http://www.broadinstitute. Discussion (A) In vitro DBF-2 activity assays after addition of the indicated kinases. MST-1-stimulated DBF-2 activity is inhibited by addition of CDC-7 or CDC-7(D195A), while SID-1-stimulation requires active CDC-7. Western blot analysis of the precipitated proteins was used to determine comparable kinase levels (n = 4). (B) A GFP fusion construct of wt CDC-7 localized to spindle pole bodies and accumulated around the mature septal pore, while a CDC7(D195A)-GFP construct exclusively localized to SPBs. Nuclei were labeled with histone H1-RFP and plasma membrane with FM4-64. doi:10.1371/journal.pgen.1004306.g005 Microscopy org/annotation/genome/neurospora/MultiHome.html) and cloned either into the pGADT7 vector containing the GAL4 activation domain or into pGBKT7 containing the DNA-binding domain. Because the pGBKT7-POD-6 fusion protein was determined to be self-activating, we generated two fragments containing the N- terminal kinase domain (aa 1–412) and C-terminal non-catalytic domain of POD-6 (aa 415–929). Fusion proteins were (co-) expressed in S. cerevisiae AH109 and potential interactions determined by growth tests on SD medium lacking the amino acids adenine, histidine, leucine and tryptophan or on SD medium supplemented with 10 mM 3-amino-1,2,4-triazole and lacking leucine, tryptophane and histidine. py Low magnification analysis of fungal hyphae or colonies was performed as described [66,67] using an SZX16 stereomicroscope, equipped with a Colorview III camera and CellD imaging software (Olympus). In order to avoid accumulation of suppressor mutations, aseptate deletion strains were maintained as hetero- karyons. An inverted Axiovert Observer Z1 microscope (Zeiss) equipped with a CSU-X1 A1 confocal scanner unit and a QuantEM 512SC camera (Photometrics) was used for spinning disk confocal microscopy [68]. Slidebook 5.0 software (Intelligent Imaging Innovations) was used for image/video acquisition, deconvolution and image analysis. Cell wall and plasma mem- brane were stained with Calcofluor White (2 mg/ml-1) and FM4- 64 (1 mg/ml-1) respectively. Time-lapse imaging was performed at capture intervals of 20–30 s for periods up to 15 min using the oil immersion objective 1006/1.3. Image series were converted into movies (*.movs). For confocal live-cell imaging, we used an inverted LSM-510 Meta laser scanning microscope (Zeiss) equipped with an argon ion laser for excitation at 488 nm wavelength and GFP filters for emission at 515–530 nm and a 636 (PH3)/1.4 N.A. oil immersion objective [69]. Laser intensity was kept to a minimum (1.5%) to reduce photobleaching and phototoxic effects. Time-lapse imaging was performed at scan intervals of 3 to 15 s for periods up to 40 min. Image resolution was 5126512 pixels and 300 dpi. Confocal images were captured using LSM-510 software and evaluated with an LSM 510 Image Examiner. Strains, media and growth conditions Strains used in this study are listed in Table S2. Genetic procedures and media used for handling N. crassa are available through the Fungal Genetic Stock Center (www.fgsc.net; [62]). April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 8 MST-1 Coordinates SIN and MOR Networks Figure 6. SID-1 and POD-6 are pathway-specific activators of the SIN and MOR, while MST-1 regulates both NDR kinase pathways. (A) In vitro kinase assays of precipitated MST-1 and SID-1 specifically stimulated DBF-2, but not DBF-2(T671A). (B) COT-1 was specifically phosphorylated by the upstream GC kinases POD-6 and MST-1, but not by SID-1. Western blot analysis of the precipitated proteins was used to determine comparable kinase levels. doi:10.1371/journal.pgen.1004306.g006 Figure 6. SID-1 and POD-6 are pathway-specific activators of the SIN and MOR, while MST-1 regulates both NDR kinase pathways. (A) In vitro kinase assays of precipitated MST-1 and SID-1 specifically stimulated DBF-2, but not DBF-2(T671A). (B) COT-1 was specifically phosphorylated by the upstream GC kinases POD-6 and MST-1, but not by SID-1. Western blot analysis of the precipitated proteins was used to determine comparable kinase levels. doi:10.1371/journal.pgen.1004306.g006 Plasmid construction and fungal expression of tagged proteins To obtain strains for the subcellular localization and expression of GFP-tagged POD-6 and HA/GFP-tagged MST-1 fusion proteins from the his-3 locus, their open reading frames were amplified by PCR as annotated by the N. crassa database using primers listed in Table S3 and introduced via SpeI/PacI, BamHI/ EcoRI or XbaI/BamHI, respectively, into pMF272 or pHAN1 [63– 65]. The kinase dead versions of CDC-7 and MST-1 were generated by site-directed mutagenesis according to the manufac- turer’s instructions (Stratagene). The final plasmids were trans- formed into his-3, nic-3;his-3 or trp-1;his-3 and were selected for complementation of the his-3 auxotrophy. Strains expressing the fusion constructs were crossed with the respective deletion strain and the progeny assayed for hygromycin resistance, GFP expression and complementation of the mutant growth/septation defects. Alternatively, histidine auxotrophic deletion strains were generated and directly transformed with plasmids allowing expression of the fusion proteins from the his-3 locus, and under control of the inducible Pccg-1 promoter. Protein methods Liquid N. crassa cultures were grown at room temperature, harvested gently by filtration using a Bu¨chner funnel and ground in liquid nitrogen. The pulverized mycelium was mixed 1:1 with IP-Buffer (50 mM Tris/HCL pH 7.5, 100 mM KCl, 10 mM MgCl2, 0.1% NP-40, 5 mM NaF, 1 mM PEFAbloc SC, 2 mM April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 9 MST-1 Coordinates SIN and MOR Networks Figure 7. MST-1/POD-6 hetero-dimerization inhibits POD-6. (A) Yeast two-hybrid tests indicate hetero-dimerization of MST-1 and POD-6. (B Co-immunoprecipitation experiments of MST-1-GFP and HA-POD-6 and of myc-COT-1 and HA-POD-6 from cell extracts co-expressing bot functionally tagged proteins indicate interaction of the two kinase pairs. However, the MST-1/POD-6 interaction was more stable and withstood tw washes with IP buffer, while the interaction between COT-1 and POD-6 was abolished under these conditions. (C) Protein displacement assay of precipitated myc-COT-1/HA-POD-6 complex by the addition of separately purified MST-1 kinase, but not a mock-IP precipitate abolished the COT- POD-6 interaction. The upper panel displays a representative experiment, while three independent experiments are quantified in the lower graph. (D POD-6 activity was determined in vitro in the presence of increasing levels of added kinase-dead MST-1(D157A) (n = 4). doi:10.1371/journal.pgen.1004306.g007 Figure 7. MST-1/POD-6 hetero-dimerization inhibits POD-6. (A) Yeast two-hybrid tests indicate hetero-dimerization of MST-1 and POD-6. (B) Co-immunoprecipitation experiments of MST-1-GFP and HA-POD-6 and of myc-COT-1 and HA-POD-6 from cell extracts co-expressing both functionally tagged proteins indicate interaction of the two kinase pairs. However, the MST-1/POD-6 interaction was more stable and withstood two washes with IP buffer, while the interaction between COT-1 and POD-6 was abolished under these conditions. (C) Protein displacement assay of a precipitated myc-COT-1/HA-POD-6 complex by the addition of separately purified MST-1 kinase, but not a mock-IP precipitate abolished the COT-1/ POD-6 interaction. The upper panel displays a representative experiment, while three independent experiments are quantified in the lower graph. (D) POD-6 activity was determined in vitro in the presence of increasing levels of added kinase-dead MST-1(D157A) (n = 4). doi:10.1371/journal.pgen.1004306.g007 DTT, 1 mM Na3VO4, 25 mM b-glycerophosphate, 2 mM benzamidine, 2 ng/ml pepstatin A, 10 ng/ml aprotinin, 10 ng/ml leupeptin) and centrifuged twice at 4uC (15 min at 4500 g and 30 min at 14000 g) to obtain crude cell extracts. Protein methods Co-immunopre- cipitation experiments were performed with cell extracts from fused, heterokaryotic strains that were selected by their ability to grow on minimal media lacking supplements. Protein-decorated beads were washed twice with IP buffer, and immunoprecipitated proteins were recovered by boiling the beads for 10 min at 98uC in 36 Laemmli buffer and separated by 10% SDS-PAGE. Mono- clonal mouse a-HA (clone HA-7, Sigma Aldrich), a-myc (9E10, Santa Cruz) and a-GFP (B2, Santa Cruz) were used in this study. Protein displacement from CDC-7-containing complexes was assayed by precipitating CDC-7-GFP from cell extracts co- expressing HA-SID-1 or HA-MST-1, respectively, and mixed with separately purified MST-1-GFP or SID-1-GFP, respec- tively. Precipitates of a mock-IP from wt extracts were used as controls. Samples were incubated for 30 min at RT and the suspensions subsequently centrifuged (2 min at 4000 g) to remove supernatant and washed once with IP buffer to remove displaced/unbound HA-SID-1. Immunoprecipitated proteins were recovered by boiling sepharose beads for 10 min at 98uC April 2014 \| Volume 10 \| Issue 4 \| e1004306 10 PLOS Genetics \| www.plosgenetics.org MST-1 Coordinates SIN and MOR Networks Figure 8. Model for the proposed interaction of the SIN and MOR networks during septation in N. crassa. See text for details. doi:10.1371/journal.pgen.1004306.g008 Figure S2 Localization of POD-6 at the hyphal tip. A GFP fusion construct of POD-6 localized at the hyphal tip in a dot-like structure in the distal region of the Spitzenko¨rper and as membrane- associated apical crescent. Plasma membrane and Spitzenko¨rper were labeled with FM4-64. (PDF) Figure S3 Dmst-1 displays synthetic interactions with SIN, but not MOR pathway mutants. (A) Dmst-16wt crosses produced a large number of round ascospores, in contrast to the typical pea- shaped ascospores generated in wt6wt crosses. Dmst-16Dmst-1 crosses were blocked after perithecium formation, resulting in fruiting bodies that lacked most asci and all ascopores. (B) Synthetic defects were observed in crosses of Dmst-1 with SIN but not MOR mutants. Dmst-16Ddbf-2 and Dmst-16Dsid-1 crosses generated empty perithecia. In contrast, Dmst-16Dcot-1 and Dmst-16Dpod-6 crosses resulted in the expected segregation of round and normally shaped ascospores. (PDF) Figure S4 Localization of the inactive kinase variants DBF- 2(D422A) and MST-1(D157A) at spindle pole bodies and constricting septa. Nuclei and plasma membrane are co-labeled with histone H1-RFP and FM4-64, respectively. (PDF) Table S1 Identification of N. crassa SIN proteins by mass spectrometry. (XLSX) Figure 8. (MOV) Video S2 4D reconstruction of z-stacks in time lapse series of the BNI-1-GFP-labeled CAR in the wt. BNI-1-GFP developed concentrically constricting rings, resulting in centrally positioned septal pores. The cell wall was stained with Calcofluor White. Images were captured at 120 s intervals. (MOV) Protein methods Model for the proposed interaction of the SIN and MOR networks during septation in N. crassa. See text for details. doi:10.1371/journal.pgen.1004306.g008 Table S2 N. crassa strains used in this study. (DOCX) Table S2 N. crassa strains used in this study. (DOCX) Table S3 Primers used in this study. (DOCX) Table S3 Primers used in this study. (DOCX) in 36 Laemmli buffer. GFP-trap experiments, mass spectrom- etry and database analysis were performed as described [68,70]. The GFP-trap pulldown reagent is a quality GFP-binding protein derived from alpaca coupled to a monovalent matrix and was obtained from Chromotek. COT-1 and DBF-2 kinase activity assays were performed as described [43,58] according to a modified protocol described previously for animal NDR kinase [71]. NDR kinase stimulation by upstream kinases was assayed as described [43]. GC kinase activity was determined by phosphorylation of the peptide substrate EESPELSLPFI- GYTFKRFDNNFR, which was used at a final concentration of 2 mM [41]. Assay conditions were analogous to NDR kinase assays described above. Video S1 Time-course of MST-1-GFP localization during septum formation. MST-1-GFP formed cortical rings at incipient septation sites that constricted during septum formation and accumulated around the septal pore of the completed septum (a) GFP channel; (b) RFP channel; (c) merged. The plasma membrane was stained with FM4-64. Images were captured at 20 sec intervals. (MOV) References Biochem Soc Trans 36: 436–438. 31. Minke PF, Lee IH, Plamann M (1999) Microscopic analysis of Neurospora ropy mutants defective in nuclear distribution. Fungal Genet Biol 28: 55–67. 10. Meitinger F, Palani S, Pereira G (2012) The power of MEN in cytokinesis. Cell Cycle 11: 219–228. 32. Kim JM, Zeng CJ, Nayak T, Shao R, Huang AC, et al. (2009) Timely septation requires SNAD-dependent spindle pole body localization of the septation initiation network components in the filamentous fungus Aspergillus nidulans. Mol Biol Cell 20: 2874–2884. 11. Weiss EL (2012) Mitotic exit and separation of mother and daughter cells. Genetics 192: 1165–1202. 12. Schneper L, Krauss A, Miyamoto R, Fang S, Broach JR (2004) The Ras/protein kinase A pathway acts in parallel with the Mob2/Cbk1 pathway to effect cell cycle progression and proper bud site selection. Eukaryot Cell 3: 108–120. 33. Rasmussen CG, Glass NL (2005) A Rho-type GTPase, rho-4, is required for septation in Neurospora crassa. Eukaryot Cell 4: 1913–1925. 34. Si H, Justa-Schuch D, Seiler S, Harris SD (2010) Regulation of septum formation by the Bud3-Rho4 GTPase module in Aspergillus nidulans. Genetics 185: 165–176. 13. Das M, Wiley DJ, Medina S, Vincent HA, Larrea M, et al. (2007) Regulation of cell diameter, For3p localization, and cell symmetry by fission yeast Rho-GAP Rga4p. Mol Biol Cell 18: 2090–2101. 35. Justa-Schuch D, Heilig Y, Richthammer C, Seiler S (2010) Septum formation is regulated by the RHO4-specific exchange factors BUD3 and RGF3 and by the landmark protein BUD4 in Neurospora crassa. Mol Microbiol 76: 220–235. 14. Vogt N, Seiler S (2008) The RHO1-specific GTPase-activating protein LRG1 regulates polar tip growth in parallel to Ndr kinase signaling in Neurospora. Mol Biol Cell 19: 4554–4569. 15. Das M, Wiley DJ, Chen X, Shah K, Verde F (2009) The conserved NDR kinase Orb6 controls polarized cell growth by spatial regulation of the small GTPase Cdc42. Curr Biol 19: 1314–1319. 36. Seiler S, Plamann M (2003) The genetic basis of cellular morphogenesis in the filamentous fungus Neurospora crassa. Mol Biol Cell 14: 4352–4364. 37. Yarden O, Plamann M, Ebbole DJ, Yanofsky C (1992) cot-1, a gene required for hyphal elongation in Neurospora crassa, encodes a protein kinase. EMBO J 11: 2159–2166. 16. Bidlingmaier S, Weiss EL, Seidel C, Drubin DG, Snyder M (2001) The Cbk1p pathway is important for polarized cell growth and cell separation in Saccharomyces cerevisiae. Mol Cell Biol 21: 2449–2462. 38. References 1. 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Fungal Biol 115: 446–474. 8. Krapp A, Simanis V (2008) An overview of the fission yeast septation initiation network (SIN). Biochem Soc Trans 36: 411–415. 30. Wolkow TD, Harris SD, Hamer JE (1996) Cytokinesis in Aspergillus nidulans is controlled by cell size, nuclear positioning and mitosis. J Cell Sci 109 (Pt 8): 2179–2188. 9. Clifford DM, Chen CT, Roberts RH, Feoktistova A, Wolfe BA, et al. (2008) The role of Cdc14 phosphatases in the control of cell division. References Seiler S, Vogt N, Ziv C, Gorovits R, Yarden O (2006) The STE20/germinal center kinase POD6 interacts with the NDR kinase COT1 and is involved in polar tip extension in Neurospora crassa. Mol Biol Cell 17: 4080–4092. 17. Mazanka E, Alexander J, Yeh BJ, Charoenpong P, Lowery DM, et al. (2008) The NDR/LATS family kinase Cbk1 directly controls transcriptional asymmetry. PLoS Biol 6: e203. 39. Maerz S, Dettmann A, Ziv C, Liu Y, Valerius O, et al. (2009) Two NDR kinase- MOB complexes function as distinct modules during septum formation and tip extension in Neurospora crassa. Mol Microbiol 74: 707–723. 18. Gutierrez-Escribano P, Zeidler U, Suarez MB, Bachellier-Bassi S, Clemente- Blanco A, et al. (2012) The NDR/LATS kinase Cbk1 controls the activity of the transcriptional regulator Bcr1 during biofilm formation in Candida albicans. PLoS Pathog 8: e1002683. 40. Richthammer C, Enseleit M, Sanchez-Leon E, Marz S, Heilig Y, et al. (2012) RHO1 and RHO2 share partially overlapping functions in the regulation of cell wall integrity and hyphal polarity in Neurospora crassa. Mol Microbiol 85: 716– 733. 19. Ray S, Kume K, Gupta S, Ge W, Balasubramanian M, et al. (2010) The mitosis- to-interphase transition is coordinated by cross talk between the SIN and MOR pathways in Schizosaccharomyces pombe. J Cell Biol 190: 793–805. 41. 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Heilig Y, Schmitt K, Seiler S (2013) Phospho-Regulation of the Neurospora crassa Septation Initiation Network. PLoS One 8: e79464. Septation Initiation Network. PLoS One 8: e79464. 23. Acknowledgments Video S6 4D reconstruction of z-stacks in time lapse series of the Lifeact-GFP-labeled CAR in Dmst-1. Failure of correct CAR assembly resulted in acentric constriction and asymmetric position of the septal pore. Images were captured at scan intervals of 0.5 to 3 s for periods up to 40 min. We thank Klaus Palme for providing lab space for the group. The Fungal Genetics Stock Center and the Neurospora program project PO1 GM068087 ‘‘Functional analysis of a model filamentous fungus’’ are acknowledged for strains. Supporting Information Figure S1 Phylogram of fungal GC kinases. The tree was generated by using ClustalX 2.1 with bootstrap support (111 random number generator seed and 1000 bootstrap trials) and the predicted protein sequences from selected ascomycete (Saccharomyces cerevisiae, Candida albicans, Ashbya gossypii, Yarrowia lipolytica, Schizo- saccharomyces pombe, Schizosaccharomyces japonicus, Neurospora crassa, Aspergillus nidulans, Magnaporthe grisea, Fusarium graminearum, Botrytis cinerea, Histoplasma capsulatum), basidiomycete (Phanerochaetae chrysos- porium, Coprinus cinereus, Ustilago maydis, Cryptococcus neoformans) and zygomycete (Rhizopus oryzae) proteins. S. cerevisiae Cdc15 was used as outgroup member (multiple alignment parameters: gap opening 50.0, gap extension 50.0, delay divergent sequences 30%, protein weight matrix gonnet series). The red, blue and yellow boxes label GC kinases with homologies to N. crassa POD-6, MST-1 and SID-1, respectively. The predicted R. oryzae protein HMPREF1544_07988 likely represents an aberrant SID-1 homolog. (PDF) Video S3 4D reconstruction of z-stacks in time lapse series of the BNI-1-GFP-labeled CAR in Dmst-1. BNI-1 formed asymmetric, open rings that led to acentric constriction and asymmetric septal pores. The cell wall was stained with Calcofluor White. Images were captured at 90 s intervals. (MOV) Video S4 Time-course of BNI-1-GFP localization during septum formation in Dmst-1. During the formation of multiple, closely spaced septa, BNI-1-GFP formed cortical, centrically and acentrically constricting rings and accumulated around the septal pore of the completed septum. Multiple, closely spaced septa can be formed in a simultaneous as well as sequential manner. The plasma membrane was stained with FM4-64. Images were captured at 20 sec intervals. (MOV) April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org PLOS Genetics \| www.plosgenetics.org 11 MST-1 Coordinates SIN and MOR Networks Video S5 4D reconstruction of z-stacks in time lapse series of the Lifeact-GFP-labeled CAR in wt cells. Lifeact-GFP labeled a mesh of F-actin cables and patches around the future septation site, which subsequently coalesced to form the CAR. Images were captured at scan intervals of 0.5 to 3 s for periods up to 40 min. (MOV) Dmst-1. Failure of correct CAR assembly resulted in the formation of open actin spirals, which were unable to constrict. Images were captured at scan intervals of 0.5 to 3 s for periods up to 40 min. (MOV) Author Contributions Video S7 4D reconstruction of z-stacks in time lapse series of the Lifeact-GFP-labeled CAR in Dmst-1. The F-actin meshwork was mis-organized and the actin cables were irregularly distributed in Conceived and designed the experiments: YH SS. Performed the experiments: YH AD RRMP KS OV. Analyzed the data: YH AD RRMP KS OV SS. Wrote the paper: YH SS. MST-1 Coordinates SIN and MOR Networks Nat Rev Microbiol 9: 876–887. 65. Kawabata T, and Inoue H (2007) Detection of physical interactions by immunoprecipitation of FLAG- and HA-tagged proteins expressed at the his-3 locus in Neurospora crassa. Fungal Genetics Newsletter 54: 5–8. g 51. Balasubramanian MK, Srinivasan R, Huang Y, Ng KH (2012) Comparing contractile apparatus-driven cytokinesis mechanisms across kingdoms. Cytoskel- eton (Hoboken) 69: 942–956. 66. Maddi A, Dettman A, Fu C, Seiler S, Free SJ (2012) WSC-1 and HAM-7 are MAK-1 MAP kinase pathway sensors required for cell wall integrity and hyphal fusion in Neurospora crassa. PLoS One 7: e42374. ( ) 52. Delgado-Alvarez DL, Callejas-Negrete OA, Gomez N, Freitag M, Roberson RW, et al. (2010) Visualization of F-actin localization and dynamics with live cell markers in Neurospora crassa. Fungal Genet Biol 47: 573–586. p 67. Maerz S, Funakoshi Y, Negishi Y, Suzuki T, Seiler S (2010) The Neurospora peptide:N-glycanase ortholog PNG1 is essential for cell polarity despite its lack of enzymatic activity. J Biol Chem 285: 2326–2332. 53. Mourino-Pe´rez R (2013) Septum development in filamentous ascomycetes. Fungal Biology Reviews 27: 1–9. 68. Dettmann A, Heilig Y, Ludwig S, Schmitt K, Illgen J, et al. (2013) HAM-2 and HAM-3 are central for the assembly of the Neurospora STRIPAK complex at the nuclear envelope and regulate nuclear accumulation of the MAP kinase MAK-1 in a MAK-2-dependent manner. Mol Microbiol 90: 796–812. 54. Rodrigo G, Elena SF (2011) Structural discrimination of robustness in transcriptional feedforward loops for pattern formation. PLoS One 6: e16904. p p p 55. Johnson AE, McCollum D, Gould KL (2012) Polar opposites: Fine-tuning cytokinesis through SIN asymmetry. Cytoskeleton (Hoboken) 69: 686–699. 56. Stegert MR, Hergovich A, Tamaskovic R, Bichsel SJ, Hemmings BA (2005) Regulation of NDR protein kinase by hydrophobic motif phosphorylation mediated by the mammalian Ste20-like kinase MST3. Mol Cell Biol 25: 11019– 11029. 69. Araujo-Palomares CL, Richthammer C, Seiler S, Castro-Longoria E (2011) Functional characterization and cellular dynamics of the CDC-42 - RAC - CDC-24 module in Neurospora crassa. PLoS One 6: e27148. 70. Riquelme M, Bredeweg EL, Callejas-Negrete O, Roberson RW, Ludwig S, et al. (2014) The Neurospora crassa exocyst complex tethers Spitzenkorper vesicles to the apical plasma membrane during polarized growth. Mol Biol Cell In press: doi: 10.1091/mbc.E13-06-0299. 57. Jansen JM, Barry MF, Yoo CK, Weiss EL (2006) Phosphoregulation of Cbk1 is critical for RAM network control of transcription and morphogenesis. J Cell Biol 175: 755–766. 58. MST-1 Coordinates SIN and MOR Networks 45. Huang TY, Markley NA, Young D (2003) Nak1, an essential germinal center (GC) kinase regulates cell morphology and growth in Schizosaccharomyces pombe. J Biol Chem 278: 991–997. 59. Emoto K, Parrish JZ, Jan LY, Jan YN (2006) The tumour suppressor Hippo acts with the NDR kinases in dendritic tiling and maintenance. Nature 443: 210– 213. J 46. Nelson B, Kurischko C, Horecka J, Mody M, Nair P, et al. (2003) RAM: a conserved signaling network that regulates Ace2p transcriptional activity and polarized morphogenesis. Mol Biol Cell 14: 3782–3803. 60. Hergovich A, Hemmings BA (2009) Mammalian NDR/LATS protein kinases in hippo tumor suppressor signaling. Biofactors 35: 338–345. pp pp g g 61. Vichalkovski A, Gresko E, Cornils H, Hergovich A, Schmitz D, et al. (2008) 61. Vichalkovski A, Gresko E, Cornils H, Hergovich A, Schmitz D, et al. (2008) NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr Biol 18: 1889–1895. g ( ) NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr Biol 18: 1889–1895. 47. Guertin DA, Chang L, Irshad F, Gould KL, McCollum D (2000) The role of the sid1p kinase and cdc14p in regulating the onset of cytokinesis in fission yeast. EMBO J 19: 1803–1815. 62. McCluskey K (2003) The Fungal Genetics Stock Center: from molds to molecules. Adv Appl Microbiol 52: 245–262. 48. Goshima T, Kume K, Koyano T, Ohya Y, Toda T, et al. (2010) Fission yeast germinal center (GC) kinase Ppk11 interacts with Pmo25 and plays an auxiliary role in concert with the morphogenesis Orb6 network (MOR) in cell morphogenesis. J Biol Chem 285: 35196–35205. 63. Freitag M, Hickey PC, Raju NB, Selker EU, Read ND (2004) GFP as a tool to analyze the organization, dynamics and function of nuclei and microtubules in Neurospora crassa. Fungal Genet Biol 41: 897–910. 49. 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April 2014 \| Volume 10 \| Issue 4 \| e1004306 April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 12 MST-1 Coordinates SIN and MOR Networks MST-1 Coordinates SIN and MOR Networks Ziv C, Kra-Oz G, Gorovits R, Marz S, Seiler S, et al. (2009) Cell elongation and branching are regulated by differential phosphorylation states of the nuclear Dbf2-related kinase COT1 in Neurospora crassa. Mol Microbiol 74: 974–989. 71. Millward TA, Heizmann CW, Schafer BW, Hemmings BA (1998) Calcium regulation of Ndr protein kinase mediated by S100 calcium-binding proteins. EMBO J 17: 5913–5922. April 2014 \| Volume 10 \| Issue 4 \| e1004306 PLOS Genetics \| www.plosgenetics.org 13
https://openalex.org/W2999962979	https://www.nature.com/articles/s41598-020-57450-8.pdf	English	null	Oropharyngeal microbiome of a college population following a meningococcal disease outbreak	Scientific reports	2,020	cc-by	8,002	OPEN Asymptomatic oropharyngeal carriage of Neisseria meningitidis peaks in adolescence and young adulthood. Following a meningococcal disease outbreak at a U.S. college, we profiled the oropharyngeal microbiomes of 158 students to identify associations between bacterial community composition and meningococcal carriage or risk factors for carriage, including male gender, smoking, and frequent social mixing. Metagenomic shotgun sequencing identified 268 bacterial taxa at the genus or species level, with Streptococcus, Veillonella, and Rothia species being most abundant. Microbiome composition showed weak associations with meningococcal carriage and risk factors for carriage. N. meningitidis abundance was positively correlated with that of Fusobacterium nucleatum, consistent with hypothesized propionic acid cross-feeding. Additional species had positive abundance correlations with N. meningitidis, including Aggregatibacter aphrophilus, Campylobacter rectus, Catonella morbi, Haemophilus haemolyticus, and Parvimonas micra. N. meningitidis abundance was negatively correlated with unidentified Veillonella species. Several of these species are commonly found in dental plaque, while N. meningitidis is primarily found in the pharynx, suggesting that ecological interactions extend throughout the oral cavity. Although risk factors for meningococcal carriage do not strongly impact most bacterial species in the oropharynx, variation in the upper respiratory tract microbiome may create conditions that are more or less favorable for N. meningitidis carriage. Neisseria meningitidis is a transient commensal of the human pharynx that also causes life-threatening diseases including meningitis and bacteremia1. Meningococcal carriage prevalence peaks in late adolescence and early adulthood2, corresponding to an increased risk of disease in this age group1. Risk factors for meningococcal carriage among adolescents and young adults in the United States include male gender, smoking, and frequent social mixing3. However, the influence of the pharyngeal microbiota on meningococcal carriage has been largely unexplored. A variety of bacterial species colonize the pharynx and other upper respiratory tract (URT) sites4,5, but there is currently little information about how they interact with N. meningitidis or how their populations are influenced by traditional risk factors for meningococcal carriage.i l y g g Understanding N. meningitidis ecology requires identification of microbial species that co-colonize the phar- ynx. Culture-based studies target particular species, often from the oropharynx, where N. meningitidis is most readily cultured6. These studies have provided evidence of competitive exclusion between N. lactamica and N. meningitidis in the oropharynx. This evidence includes the low prevalence of N. lactamica carriage among age groups with highest prevalence of N. meningitidis carriage7,8, and the reduced probability of isolating N. www.nature.com/scientificreports www.nature.com/scientificreports www.nature.com/scientificreports Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 OPEN Characteristics of oropharyngeal swabs selected for shotgun metagenomic sequencing. aSmoking in the past 30 days. bNumber of visits to bars, clubs, or parties per week. cThe swabs available for sequencing were limited to students who were between the age of 18 and 23 years, had received both a quadrivalent meningococcal conjugate vaccine (MenACWY) and at least two doses of MenB-FHbp vaccine at least two weeks prior to specimen collection, and had not used antibiotics in the past 30 days. Porphyromonas. Further analysis of 16S rRNA genes showed that the average proportional abundance of both Fusobacterium spp. and Porphyromonas spp. increases through adolescence, in parallel to meningococcal carriage in the nasopharynx11.fi Interpretation of 16S rRNA gene sequences can be limited by difficulty discriminating between N. mening- itidis and other Neisseria species12,13. However, shotgun metagenomic sequencing can readily discriminate among the Neisseria species that are present in different parts of the oral cavity14. Furthermore, shotgun metagenomic sequencing enables identification of genes encoding metabolic enzymes in a bacterial community15.l q gi g g y y To investigate how carriage of N. meningitidis among young adults may be influenced by variation in the bac- terial community, we performed metagenomic shotgun sequencing on DNA obtained from oropharyngeal swabs collected from college students following a serogroup B meningococcal (MenB) disease outbreak in Providence, Rhode Island3. We evaluated the proportional abundances of each bacterial species in the oropharynx and identi- fied species that were associated with meningococcal carriage or risk factors for meningococcal carriage. OPEN mening- itidis from oropharyngeal swabs following inoculation with N. lactamica9. Sequence-based, culture-independent approaches provide a more comprehensive assessment of the oro- pharyngeal bacterial community10, enabling both the discovery of unanticipated relationships and the evaluation of new hypotheses using previously collected data. For instance, N. meningitidis can utilize propionic acid as a carbon source, prompting the hypothesis that N. meningitidis growth is promoted by the presence of anaerobic bacteria that produce this short fatty acid by fermentation11. This cross-feeding hypothesis was initially evalu- ated with 16S rRNA gene data from bacteria in salivary specimens, identifying correlations between the propor- tional abundances of Neisseria spp. and two genera of anaerobic propionic acid producers, Fusobacterium and 1Division of Bacterial Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA. 2Present address: Division of Scientific Education and Professional Development, Centers for Disease Control and Prevention, Atlanta, GA, USA. *email: ymw8@cdc.gov; gqe8@cdc. gov Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ Swab characteristic Swab counts N. meningitidis culture results Smoking statusa Social mixing per weekb Gender Sequenced (% of 158) Availablec (% of 705) Positive Smoker ≥1 Male 12 (7.6%) 47 (6.7%) Positive Smoker ≥1 Female 14 (8.9%) 19 (2.7%) Positive Smoker <1 Male 3 (1.9%) 3 (0.4%) Positive Smoker <1 Female 1 (0.6%) 1 (0.1%) Positive Non-smoker ≥1 Male 10 (6.3%) 23 (3.3%) Positive Non-smoker ≥1 Female 14 (8.9%) 44 (6.2%) Positive Non-smoker <1 Male 12 (7.6%) 12 (1.7%) Positive Non-smoker <1 Female 9 (5.7%) 12 (1.7%) Negative Smoker ≥1 Male 12 (7.6%) 78 (11.1%) Negative Smoker ≥1 Female 11 (7.0%) 46 (6.5%) Negative Smoker <1 Male 9 (5.7%) 17 (2.4%) Negative Smoker <1 Female 9 (5.7%) 10 (1.4%) Negative Non-smoker ≥1 Male 9 (5.7%) 59 (8.4%) Negative Non-smoker ≥1 Female 11 (7.0%) 164 (23.3%) Negative Non-smoker <1 Male 10 (6.3%) 51 (7.2%) Negative Non-smoker <1 Female 12 (7.6%) 119 (16.9%) Total 158 (100%) 705 (100%) Table 1. Characteristics of oropharyngeal swabs selected for shotgun metagenomic sequen in the past 30 days. bNumber of visits to bars, clubs, or parties per week. cThe swabs availab were limited to students who were between the age of 18 and 23 years, had received both a meningococcal conjugate vaccine (MenACWY) and at least two doses of MenB-FHbp vacc weeks prior to specimen collection, and had not used antibiotics in the past 30 days. Table 1. Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 Results h Cells are color coded on a logarithmic scale to show the proportional abundance of each species (row) in each swab (column). In both panels, swabs are i Figure 1. Bacterial species composition of 158 oropharyngeal swabs, clustered by species composition. (A) Abundance of each genus that accounts for over 10% of any swab. Average abundance is in parentheses in legend. (B) Heatmap showing the 100 most common bacterial taxa. Cells are color coded on a logarithmic scale to show the proportional abundance of each species (row) in each swab (column). In both panels, swabs are clustered according to average Bray-Curtis distances of their taxonomic profiles, represented by the dendrogram on the top. Taxa in panel B are clustered according to the Bray-Curtis distance of their abundance in each swab, shown by the dendrogram on the left. Colored portions of the dendrogram indicate groups for which distances are <70% of the maximum distance. Neisseria meningitidis is emphasized with bold font. Figure 1. Bacterial species composition of 158 oropharyngeal swabs, clustered by species composition. (A) Abundance of each genus that accounts for over 10% of any swab. Average abundance is in parentheses in legend. (B) Heatmap showing the 100 most common bacterial taxa. Cells are color coded on a logarithmic scale to show the proportional abundance of each species (row) in each swab (column). In both panels, swabs are clustered according to average Bray-Curtis distances of their taxonomic profiles, represented by the dendrogram on the top. Taxa in panel B are clustered according to the Bray-Curtis distance of their abundance in each swab, shown by the dendrogram on the left. Colored portions of the dendrogram indicate groups for which distances are <70% of the maximum distance. Neisseria meningitidis is emphasized with bold font. Figure 1. Bacterial species composition of 158 oropharyngeal swabs, clustered by species composition. (A) Abundance of each genus that accounts for over 10% of any swab. Average abundance is in parentheses in legend. (B) Heatmap showing the 100 most common bacterial taxa. Cells are color coded on a logarithmic scale to show the proportional abundance of each species (row) in each swab (column). In both panels, swabs are clustered according to average Bray-Curtis distances of their taxonomic profiles, represented by the dendrogram on the top. Taxa in panel B are clustered according to the Bray-Curtis distance of their abundance in each swab, shown by the dendrogram on the left. Results h The oropharynx harbors a diverse bacterial community. Following four cross-sectional oropharyn- geal carriage surveys of undergraduate students, 158 oropharyngeal swabs were selected for sequencing from a pool of 705 swabs collected during the third and fourth rounds from students who were between the age of 18 and 23 years, had not used antibiotics in the past 30 days, and had received both a quadrivalent meningococcal conjugate vaccine (MenACWY) and at least two doses of MenB-FHbp vaccine at least two weeks prior to swab collection. The 158 oropharyngeal swabs included 75 N. meningitidis culture-positive swabs (64 nongroupable, 6 serogroup E, and 5 serogroup B). The culture-positive and culture-negative swabs had similar composition with respect to the students’ carriage risk factors: gender, smoking, and frequent social mixing (Table 1; see methods for details).i A total of 268 bacterial taxa were identified among the metagenomic sequences from the 158 oropharyngeal swabs; 239 of these taxa are species that are represented in the reference genome database, while 29 of the taxa are “unclassified” species belonging to different genera. The total proportional abundance of unclassified species ranges from 1.4% to 46.2% of the bacterial community (mean of 19.5%). The proportional abundance of taxa varied substantially across specimens (Fig. 1). While five genera (Streptococcus, Veillonella, Rothia¸ Prevotella, and Actinomyces) were found in all 158 specimens, their propor- tional abundance varied by orders of magnitude between specimens (Fig. 1A). For instance, the most abundant genus, Streptococcus, ranged from 2.4% to 66%. Three species from two genera were detected in all 158 speci- mens: Rothia mucilaginosa, Streptococcus salivarius, and Streptococcus parasanguinis (Fig. 2). Clustering of swabs according to bacterial community composition placed 135 (85%) into a single group (Fig. 1, Supplemental Fig. 1). The effective number of taxa (exponential Shannon entropy) within the oropharyngeal specimens (13.4) is within the range of values calculated from taxonomic profiles of oral body sites generated by the Human Microbiome Project (HMP) using comparable methods (Supplemental Table 1). The taxonomic composition is most similar to that of the tongue dorsum (Supplemental Figs. 2 and 3; Mann-Whitney U, p < 0.001). Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ www.nature.com/scientificreports ature.com/scientificreports/ Figure 1. Bacterial species composition of 158 oropharyngeal swabs, clustered by species composition. (A) Abundance of each genus that accounts for over 10% of any swab. Average abundance is in parentheses in legend. (B) Heatmap showing the 100 most common bacterial taxa. Results h meningitidis culture Negative (83) Positive (75) 1.320 [1.200, 1.499] 1.326 [1.203, 1.509] 0.08 Social mixing per weekb <1 (65) ≥1 (93) 1.326 [1.202, 1.515] 1.320 [1.201, 1.496] 0.93 Gender Male (77) Female (81) 1.319 [1.198, 1.500] 1.328 [1.205, 1.509] 0.02 Smokerc No (87) Yes (71) 1.318 [1.200, 1.501] 1.328 [1.204, 1.508] 0.05 URT infectiond No (119) Yes (39) 1.326 [1.200, 1.512] 1.319 [1.204, 1.492] 0.91 Study round Sep 2015 (78) Mar 2016 (80) 1.323 [1.202, 1.506] 1.323 [1.201, 1.505] 0.55 Table 2. Median pairwise bacterial community diversity, within and between categories. aEffective number of communities based on Shannon entropy was measured for all 12,403 possible pairs of 158 swabs, with each pair assigned to “within” or “between” category depending on whether the two swabs were from the same or different category. The first and third quartiles are noted in brackets. bNumber of visits to bars, clubs, or parties per week. cSmoking in the past 30 days. dSymptoms of URT infection in past 2 weeks. eMann-Whitney U one- tailed p-value; a value < 0.0083 is required attain a Bonferroni-corrected significance of 0.05. Table 2. Median pairwise bacterial community diversity, within and between categories. aEffective number of communities based on Shannon entropy was measured for all 12,403 possible pairs of 158 swabs, with each pair assigned to “within” or “between” category depending on whether the two swabs were from the same or different category. The first and third quartiles are noted in brackets. bNumber of visits to bars, clubs, or parties per week. cSmoking in the past 30 days. dSymptoms of URT infection in past 2 weeks. eMann-Whitney U one- tailed p-value; a value < 0.0083 is required attain a Bonferroni-corrected significance of 0.05. Likewise, only two significant correlations were identified between the proportional abundance of each taxon and variables describing the oropharyngeal specimen, using a multivariate linear model with a false discovery limit of 25%. The proportional abundance of N. meningitidis in the metagenomic data was significantly higher among swabs that were positive for N. meningitidis by culture (p = 3.4 × 10−4); the median proportional abun- dance was 3.57 × 10−4 in the Nm culture-negative swabs (99% CI, 1.72 × 10−4–9.65 × 10−4) and 1.48 × 10−3 in the Nm culture-positive swabs (99% CI, 4.92 × 10−4–3.98 × 10−3). S. Results h Colored portions of the dendrogram indicate groups for which distances are <70% of the maximum distance. Neisseria meningitidis is emphasized with bold font. Bacterial community composition is not associated with student traits. We tested whether the bacterial community differed between swabs based on N. meningitidis culture results, three carriage risk factors (the student’s gender, smoking behavior, and social mixing behavior), and two other potentially influential factors (the student having symptoms of URT infection in the prior two weeks, and the study round in which the swab was collected). The effective number of communities (exponential Shannon beta diversity) was no lower for pairs of swabs with the same values for any of these variables than for pairs of swabs with different values (Table 2, Supplemental Figs. 1 and 4). None of the six tests passed the 5% Bonferroni family-wise error rate, which required uncorrected p-values to be less than 0.83%. Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ Figure 2. Abundance of dominant species among 158 oropharyngeal swabs. Curves show number of swabs in which each species was found at the given abundance; the y-intercept is equal to the number of swabs in which the species was identified. The twenty species shown have an abundance greater than 10% in one or more specimens, and a mean abundance greater than 1% overall (shown in parentheses in legend). Figure 2. Abundance of dominant species among 158 oropharyngeal swabs. Curves show number of swabs in which each species was found at the given abundance; the y-intercept is equal to the number of swabs in which the species was identified. The twenty species shown have an abundance greater than 10% in one or more specimens, and a mean abundance greater than 1% overall (shown in parentheses in legend). Characteristic Categories, Value (N) Median [Q1, Q3] of pairwise beta diversitya pe Within the same category Between categories N. Discussion b d To better understand the ecology of N. meningitidis, we examined the oropharyngeal microbiome among young adults at a U.S. college where 20%–24% of surveyed students carried meningococcal bacteria, as detected via culture3. Previous DNA-based profiling of the oropharyngeal microbiome has used amplicon sequencing of 16S rRNA genes for genus-level analysis of bacterial communities4,17,18. The major oropharyngeal bacterial genera identified here among college students were similar to those described previously among adult populations such as United States active duty military personnel17 and a portion of the German National Cohort study18. The most prominent differences were a high abundance of Rothia spp. and a low abundance of Leptotrichia spp. among this college population. Shotgun metagenomic sequencing obtains species-level resolution for most of the bacterial community, but published shotgun metagenomic sequence data for the oropharyngeal microbiome is limited; comparison of these oropharyngeal communities’ profiles to the profiles published by the Human Microbiome Project showed that they were most similar to the communities of the tongue dorsum.hi The overall bacterial community profile was not associated with any of the meningococcal carriage risk fac- tors that we tested. Only one bacterial species was associated with any student trait tested here; S. salivarius was detected in all swabs but was more abundant among females. While we did not identify any association between smoking and bacterial taxon abundance, previous research has found that tobacco smoking is associated with differences in bacterial species abundance in oral rinse19 and nasopharyngeal20 specimens. These studies dif- fered from our analysis of oropharyngeal specimens in several respects, including the examination of different body sites, older populations, and 16S rRNA gene sequencing rather than shotgun metagenomic sequencing. Furthermore, the oral rinse microbiome study of Bornigen et al.19 specified the intensity of cigarette use, finding that most of the differences in taxon abundance were primarily due to the comparison between regular cigarette smokers and cigarette non-smokers. In contrast, our smoking variable included regular cigarette smokers, occa- sional cigarette smokers (in the past 30 days), and marijuana smokers. The inclusion of a broader set of behaviors as “smoking” may have diminished the apparent association between smoking and bacterial taxon abundance. Patterns of co-occurrence among microbial species can provide insights into ecological relationships, such as competition and mutualism21. Among the proposed ecological relationships of N. meningitidis is competition with N. lactamica in the pharynx, resulting in lower meningococcal carriage among individuals colonized with N. lactamica9. www.nature.com/scientificreports/ www.nature.com/scientificreports/ Taxon Pearson r (p) Spearman r (p) SparCC (p) Positive correlation Aggregatibacter aphrophilus 0.22 (5.6 × 10−3) 0.17 (3.6 × 10−2) 0.10 (6 × 10−3) Campylobacter rectus 0.31 (7.4 × 10−5) 0.27 (5.4 × 10−4) 0.15 (<1 × 10−3) Catonella morbi 0.26 (8.4 × 10−4) 0.22 (4.8 × 10−3) 0.08 (7 × 10−3) Haemophilus aegyptius 0.21 (9.7 × 10−3) 0.25 (1.7 × 10−3) 0.08 (3 × 10−3) Haemophilus haemolyticus 0.20 (1.1 × 10−2) 0.37 (1.7 × 10−6) 0.17 (<1 × 10−3) Parvimonas micra 0.29 (2.5 × 10−4) 0.25 (1.7 × 10−3) 0.16 (<1 × 10−3) Negative correlation Veillonella unclassified −0.22 (5.3 × 10−3) −0.25 (1.7 × 10−3) −0.35 (<1 × 10−3) Table 3. Bacterial taxa whose proportional abundance correlates to that of N. meningitides. Note: p-values are not corrected for multiple hypothesis testing. Listed species had False Discovery Rate ≤25% for each of the three tests shown. Table 3. Bacterial taxa whose proportional abundance correlates to that of N. meningitides. Note: p-values are not corrected for multiple hypothesis testing. Listed species had False Discovery Rate ≤25% for each of the three tests shown. species16; the Pearson correlation of H. influenzae and N. meningitidis abundances was not significant (r = 0.06; p = 0.46), but the Spearman (r = 0.34, p = 1.0 × 10−5) and compositional correlations (r = 0.17, p = 0.004) were significant. The only taxon for which all three tests identified a significant negative correlation with N. mening- itidis was unclassified Veillonella, representing one or more Veillonella spp. that could not be identified at the spe- cies level. Notably, N. lactamica did not have a significant correlation with N. meningitidis (Supplemental Table 2). Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 Results h salivarius was associated with females (p = 3.8 × 10−4); the median abundance was 6.0 × 10−2 among males (99% CI, 4.2 × 10−2–8.7 × 10−2) and 9.8 × 10−2 among females (99% CI, 7.4 × 10−2–1.7 × 10−1). No associations were identified with the remaining variables (study round, URT infections, smoking, or social mixing). ‘N. meningitidis’ abundance is correlated with the propionic acid producer, Fusobacterium nucleatum. Catanazzi et al. proposed that N. meningitidis growth in the oral cavity is promoted by the pres- ence Fusobacterium spp. or Porphyromonas spp., which produce propionic acid by anaerobic fermentation11. Of the Fusobacterium or Porphyromonas species detected by metagenomic sequencing, only F. nucleatum had a significant compositional correlation with N. meningitidis (r = 0.22, p < 0.001). None of the remaining three Fusobacterium species (F. gonidiaformans, F. necrophorum, F. periodonticum) nor the four Porphyromonas taxa (P. catoniae, P. endodontalis, P. gingivalis, Porphyromonas oral taxon 279) had statistically significant compositional correlations with N. meningitidis (Supplemental Table 2). g ( pp ) Functional analysis of gene sequences in the metagenomic data indicated the potential for propionic acid production through the L-1,2-propanediol degradation pathway; no other pathway for fermentation to propionic acid was confidently identified. The L-1,2-propanediol degradation pathway was identified in 57 specimens; it was attributed to F. nucleatum in 18 specimens, to F. periodonticum in 27 specimens, and to both F. nucleatum and F. periodonticum in 12 specimens. The normalized abundance of genes for the L-1,2-propanediol degradation pathway was not correlated with the proportional abundance of N. meningitidis (Spearman r = −0.11, p = 0.16). ‘N. meningitidis’ abundance is correlated with other species. To identify potential interactions between N. meningitidis and any of the 267 taxa, we used three different correlation coefficients (Pearson’s, Spearman’s, and compositional) to test for correlations in proportional abundance between taxa. Six different taxa had significant positive correlations with N. meningitidis according to all three tests: Aggregatibacter aphrophilus, Campylobacter rectus, Catonella morbi, Haemophilus aegyptius, Haemophilus haemolyticus, and Parvimonas micra (Table 3). Haemophilus aegyptius is commonly described as a biogroup of H. influenzae rather than a separate Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ plaque and the gingival crevice, particularly A. aphrophilus, C. rectus, and C. morbi25–28; H. haemolyticus is also commonly found in dental plaque, as well as the pharynx29. y p q p y Veillonella species (for which a negative correlation with N. meningitidis was found) are commonly found in dental plaque, buccal mucosa, and the tongue26. Of note, like members of the genera Fusobacterium and Porphyromonas, members of Veillonella are anaerobic producers of propionic acid. While many Veillonella genomes encode genes for fermentation to propionic acid (e.g. V. atypica)30, these genes were not confidently identified in our metagenomic data. In contrast to members of Fusobacterium and Porphyromonas, members of Veillonella are abundant in the oral cavity throughout life26 and have been found to be more abundant among younger adolescents than older adolescents31.hii y g The significant correlations identified between the proportional abundances of N. meningitidis and other bac- terial species suggests possible beneficial or antagonistic interactions between these species and N. meningitidis. However, the bacterial species abundance is also affected by factors that were not examined in this study, such as oral and gastrointestinal health19,32. Such unmeasured and untested factors could influence the correlations between the abundances of N. meningitidis and other species. For instance, the negative correlation between N. meningitidis and Veillonella abundances could reflect the tendency for their abundances to change with age. gl y g g As sequencing technology and associated analysis tools advance, expanded investigation of URT ecology will be possible, providing context for studies of N. meningitidis transmission and host interactions. To understand meningococcal carriage and disease, further investigations will need to incorporate longitudinal observations of the microbiome as N. meningitidis carriage is gained or lost, particularly alongside medical interventions such as vaccination and antimicrobial treatments. Sampling from additional locations of the URT5 may help explain the correlations that we detected among the abundances of certain bacterial species that typically grow in den- tal plaque and the abundance of N. meningitidis. Examination of the nasopharynx is also important, since it is known to harbor N. meningitidis6, yet its bacterial community is dominated by different genera than the oro- pharynx (e.g. Staphylococcus, Corynebacterium, and Moraxella17). Further microbiome profiling can identify the large-scale structure of microbial communities, and guide more detailed studies to evaluate bacterial physiology and micron-scale interactions in situ. Methods S i Specimen collection. Oropharyngeal swabs were collected during four cross-sectional carriage surveys performed alongside a MenB-FHbp (factor H binding protein) vaccination campaign in response to the out- break, as described by Soeters, et al.3. A total of 1,121 undergraduate students participated in the third and fourth study rounds (September 2015 and March 2016), during which one prong of a bifurcated swab was preserved for sequencing in 1 ml skim milk/tryptone/glucose/glycerin medium (STGG) on ice33, then frozen at −80 °C. The other prong was used for culture-based detection of N. meningitidis by plating on a modified Thayer-Martin (MTM) agar plate (BD BBL, Franklin Lakes, New Jersey) that was incubated at 37 °C with 5% CO2 and checked for growth at 24, 48, and 72 hours3. Colonies with typical Neisseria morphology were subcultured onto a blood agar plate (BD BBL), and N. meningitidis cultures were identified using Gram staining (BD BBL), oxidase test- ing (Hardy Diagnostics, Santa Maria, California), sodC rt-PCR, and API Neisseria-Haemophilus strip testing (bioMerieux, Durham, North Carolina)34. N. meningitidis isolates were serogrouped by slide agglutination34. Participants completed a short, self-administered questionnaire including potential meningococcal carriage risk factors, including gender, smoking (tobacco or marijuana in the past 30 days), frequent social mixing (visits to bars, clubs, or parties at least once per week), and recent URT infections (symptoms in the previous two weeks). Th l 3 d b h h d l d f l h l d The carriage evaluation3 was approved by the Rhode Island Department of Health Institutional Review Board (IRB) (#IRB 2015 – 02) and the Providence College IRB (#15-062), and was determined to be public health non-research by the CDC National Center for Immunization and Respiratory Diseases (#2015 6436) and did not require CDC IRB approval. All participants provided written informed consent to participate in the carriage evaluation3 and for their oropharyngeal specimens to be stored for future research. This microbiome study used de-identified specimens and questionnaire data and was determined to not involve human subjects (#2016 6651) and did not require CDC IRB approval. DNA extraction and sequencing. DNA was extracted from 200 μL of STGG medium using QIAamp DNA Microbiome Kit (Qiagen) as per manufacturer protocol. DNA concentrations were measured using a Qubit 2.0 fluorometer with dsDNA High Sensitivity reagents (Invitrogen, Grand Island, NY, USA). The eluted DNA con- centration ranged from 0.2 to 5 ng/μL. www.nature.com/scientificreports/ Ultimately, understanding the URT microbiome will improve knowledge for a variety of diseases, ranging from oral cancer19 to bacterial infections4. Discussion b d In this study, no significant association between N. lactamica and N. meningitidis abundances was identified. This is perhaps due to the low prevalence of N. lactamica carriage among older adolescents7,8, which was reflected in the detection of N. lactamica in only 18 of the 158 swabs. Ultimately, the metagenomic data indicate that competition with N. lactamica was not a substantial contributor to the variation in N. meningitidis carriage within this population. N. meningitidis is capable of using propionic acid as a carbon source, and therefore may benefit from cross-feeding with propionic acid producers11. Studies based on 16S rRNA gene sequencing have shown that in saliva, the average proportional abundance of the anaerobic propionic acid producing genera Fusobacterium and Porphyromonas increases through adolescence, in parallel to meningococcal carriage in the nasopharynx11. The metagenomic data of the current study improved on prior 16S rRNA gene studies by estimating species-level abundances for all three genera, thereby identifying a significant positive correlation between N. meningitidis and F. nucleatum. The metagenomic data also confirmed the presence of F. nucleatum genes encoding a fermentation pathway that produces propionic acid; however, the abundance of genes for this pathway within the full microbial community was not correlated with N. meningitidis abundance.f Interactions between F. nucleatum and N. meningitidis may be limited by their tendency to grow in different locations within the URT – the gingival crevice for F. nucleatum22 and the pharynx for N. meningitidis. However, interactions may be mediated by saliva23, in which propionic acid levels can reach 3.3 mM24. The correlations in our metagenomic data suggest that N. meningitidis may interact with other taxa that are thought to grow in dental www.nature.com/scientificreports/ Methods S i Sequencing libraries were prepared from 2 ng of DNA using Nextera XT DNA library preparation kit follow- ing the manufacturer’s protocol (Illumina, San Diego, CA, USA). The resulting libraries were sequenced on three Rapid-Runs of an Illumina HiSeq 2500 (CDC Genomic Core facility) to obtain 250 bp paired-ends reads. Sixteen specimens with exceptionally low numbers of reads (<100,000 reads) were sequenced again on an Illumina MiSeq using a fresh library preparation. Metagenomic sequence analysis. Human DNA was removed by mapping reads to the hg38 reference genome using Bowtie2 (v2.2.9; local alignment mode)35, retaining a median of 94.8% (interquartile range: 85.7–98.4%) of reads with a minimum of 336,841 read pairs per specimen. Unmapped reads were submitted to SRA BioProject PRJNA428766. PCR duplicates were removed with bbtools-clumpify (v37.41)36, then Nextera adapter sequences and low-quality base calls were trimmed from reads with cutadapt (v1.8.3; options n = 5, trim-n = True, m = 50)37. The proportional abundance of bacterial species was estimated using MetaPhlAn2 (v2.2.0)38, which is a precise method for species detection (i.e. it rarely reports the presence of species that are not in a specimen)39. This precision is achieved by identifying marker genes that clearly distinguish different species, Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ and if a sufficient number of these marker genes cannot be detected to confidently identify a species, MetaPhlAn2 conservatively limits its reporting to the genus or a group of species (e.g., Streptococcus mitis/oralis/pneumoniae). Data from the Human Microbiome Project (HMP) was retrieved as MetaPhlAn2 abundance files from the HMP website40. Community taxonomic diversity of both the HMP specimens and the specimens from this study was calcu- lated from MetaPhlAn2 taxonomic profiles using an exponential transformation of Shannon entropy to describe the effective number of taxa within a specimen (alpha) and the effective number of distinct communities in a collection of specimens (beta)41. Alpha diversity approaches a lower limit of 1 when a single species dominates the community; alpha diversity is equal to the number of observed taxa in a specimen when taxa are equally abundant in a specimen. Beta diversity approaches a lower limit of 1 when specimens have identical taxonomic profiles; beta diversity equals the number of specimens in the collection when there are no shared taxa between the specimens. Data availabilityh y The dataset supporting the conclusions of this article is available in the Short Read Archive repository, BioPro PRJNA428766. y The dataset supporting the conclusions of this article is available in the Short Read Archive repository, BioProject RJNA428766. Methods S i p Associations of taxon abundance with specimen characteristics were calculated with MaAsLin (v0.0.4, default parameters)42, which applies a multivariate linear model to associate the abundance of each taxon with the avail- able metadata. MaAsLin adjusts for multiple hypothesis testing by first applying a Bonferroni correction for each taxon’s association with each value of each metadata variable, then calculating a False Discovery Rate (FDR) over all taxa and metadata variables, reporting associations with FDR ≤ 25%. Metadata variables included in the mul- tivariate linear model were: study round; N. meningitidis culture results; gender; smoking; social mixing; URT infection; and sequencing run. q g Compositional correlations calculated with SparCC (v0.1)43 were used to assess correlated taxon abundances. Calculations were performed using pseudo-counts that were generated by multiplying proportional abun- dances by 1,000; this is a conservative scaling factor, as the proportional abundance table for each specimen was generated from at least 336,841 non-human read pairs and contained at least one taxon with proportional abundance under 0.001. Tests for previously predicted correlations between N. meningitidis and Fusobacterium spp. or Porphyromonas spp. were considered significant if the compositional correlation was more extreme than 99.375% of the correlation coefficients from permutated matrices (two-tailed, 1,000 permutations; Bonferroni adjustment for 8 tests with α ≤ 5%). When testing for associations that had not been previously hypothesized, a Benjamini-Hochberg FDR44 was applied to account for the large number of comparisons (267), and specificity was increased by using both Pearson and Spearman correlations to confirm associations (two-tailed t-test)21, controlling for FDR ≤ 25% for each test. g A functional interpretation of metagenome data was performed on each specimen using HUMAnN2 v2.915 (default parameters, UniRef90 database). The potential for propionic acid production was assessed based on the presence of genes from each of the five pathways within the “Fermentation to Propanoate” class on the MetaCyc website30: “L-1,2-propanediol degradation pathway” (PWY-7013), “L-alanine fermentation to propanoate and acetate” (PROPFERM-PWY), “L-glutamate degradation VIII” (PWY-5088), and “pyruvate fermentation to pro- panoate” (P108-PWY and PWY-5494). The abundance of each pathway was measured at the community level and normalized to “copies per million” reads using the human2_renorm_table script. Confident detection of pathways required the coverage value to exceed 0.5. q g Data analysis was performed with SciPy45 with scikit-bio version 0.5.2. References 1. MacNeil, J. R., Blain, A. E., Wang, X. & Cohn, A. C. Current epidemiology and trends in meningococcal disease-United States 1996–2015. Clin Infect Dis 66, 1276–1281, https://doi.org/10.1093/cid/cix993 (2018). 1. MacNeil, J. R., Blain, A. E., Wang, X. & Cohn, A. 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Acknowledgements We would like to acknowledge assistance and advice provided by Sandeep Joseph, the CDC Biotechnology Core Facility, and the Rhode Island Meningococcal Carriage Evaluation Team. This work was supported by the Advanced Molecular Detection initiative at the Centers for Disease Control and Prevention. The funders of this research had no role in the design of the study, the collection, analysis, and interpretation of data, or in writing the manuscript. The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8 www.nature.com/scientificreports/ Author contributions A.C.R., C.B.K., L.D.R.R., A.C., H.M.S., M.J.W. and X.W. participated in the conception and design of thi manuscript; C.B.K., L.D.R.R., H.M.S. and M.J.W. participated in the acquisition of the data; A.C.R., C.B.K., A.C and X.W. participated in the analysis and interpretation of the data; A.C.R. wrote the manuscript; all author provided critical review of the manuscript and approved its publication. Competing interests The authors declare no competing interests. Additional information Supplementary information is available for this paper at https://doi.org/10.1038/s41598-020-57450-8. Correspondence and requests for materials should be addressed to A.C.R. or X.W. Reprints and permissions information is available at www.nature.com/reprints. 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To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2020 Author contributions A.C.R., C.B.K., L.D.R.R., A.C., H.M.S., M.J.W. and X.W. participated in the conception and design of this manuscript; C.B.K., L.D.R.R., H.M.S. and M.J.W. participated in the acquisition of the data; A.C.R., C.B.K., A.C. and X.W. participated in the analysis and interpretation of the data; A.C.R. wrote the manuscript; all authors provided critical review of the manuscript and approved its publication. Additional information Supplementary information is available for this paper at https://doi.org/10.1038/s41598-020-57450-8. Correspondence and requests for materials should be addressed to A.C.R. or X.W. Correspondence and requests for materials should be addressed to A.C.R. or X.W. Reprints and permissions information is available at www.nature.com/reprints. Reprints and permissions information is available at www.nature.com/reprints. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre- ative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not per- mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Cre- ative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not per- mitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. © The Author(s) 2020 Scientific Reports \| (2020) 10:632 \| https://doi.org/10.1038/s41598-020-57450-8
https://openalex.org/W4310932682	https://zenodo.org/record/7417273/files/SSPME0704.pdf	Kinyarwanda	null	FAMILISTIK KOMPETENTLIKNING YOSHLAR TARBIYASIDA OILAVIY MADANIYATNI SHAKLLANTIRISHDAGI AHAMIYATI	Zenodo (CERN European Organization for Nuclear Research)	2,022	cc-by	1,087	Nizomiy nomidagi toshkent davlat pedagogika universiteti magistranti https://doi.org/10.5281/zenodo.7417273 Nizomiy nomidagi toshkent davlat pedagogika universiteti magistranti https://doi.org/10.5281/zenodo.7417273 Annotatsiya: Har qanday tarixiy davrning ijtimoiy hayoti, albatta, oila, oilaviy munosabatlar, yosh avlod tarbiyasini butun jamiyat taraqqiyotining asosiy vazifalarining eng muhim bo‘g‘ini deb hisoblagan. Davrlar o‘zgardi, insoniyatning o‘zi o‘zgardi, oilaga munosabat ham o‘zgardi, familistik madaniyat shakllandi. Shakllanish davlat va oilani o‘zaro izolyatsiya qilishdan boshlab, davlat, davlat siyosati doirasida amalga oshirildi va natijada eng yaqin hamkorlikka, o‘zaro birlikni tan olishga erishildi. Kalit so`zlar: Familistik madaniyat, oilaviy munosabatlar, pedagog, ma’naviy- ruhiy psixologik muhit, Ommaviy madaniyat, ijtimoiy meros familistik kompetentlik Zamonaviy jamiyat taraqqiyotining mantiqiyligi zamonaviy madaniyatda oila haqidagi g‘oyalarning rivojlanishi, ommaviy ongda oila qiyofasining o‘zgarishi bilan belgilanadi. Madaniy o‘zgarishlar jarayoni alohida ahamiyatga ega bo‘lib, uning davomida madaniyat va jamiyatni yanada rivojlantirish uchun asoslarni izlash ustuvor vazifalardan biridir. Oila va oila ichidagi munosabatlarni har qanday yozuvchi ijodidagi eng muhim syujet tashkil etuvchi lahzalardan biri deb atash mumkin. Adabiyotda otalar va bolalar mavzusini, oila ichidagi munosabatlarni, o‘zini yangi voqelikdan qidirishni aks ettirmaydigan bunday asarlar kam emas. Ko‘plab badiiy so‘z ustalari jamiyat va inson muammolarini hayotning ijtimoiy sohasi prizmasidan ko‘rib chiqdilar, chunki oila shaxsning eng yaqin bebaho mikrokosmosidir. Har qanday jamiyat taraqqiyoti oiladan boshlanadi. Yosh avlodning o‘ta jiddiy qadamga tayyor bo‘lishi, yaqinlari oldidagi mas’uliyat, o‘ziga xos xususiyatga ega mustahkam oila, oilaviy munosabatlar madaniyatini shakllantirish – bu jamiyatning barcha tuzilmalarining uzoq, mashaqqatli mehnatidir. Bu ishni qachon va kim bilan boshlash kerak? Turli xil yondashuvlar mumkin, lekin bir narsani tushunish va o‘zlashtirish muhim: jamiyatning ijtimoiy rivojlanishidagi bitta bo‘g‘inni o‘tkazib yubora olmaysiz. Inson kamolotining har bir bosqichi maxsus bilim, tushunish, familistik madaniyatni idrok etish bilan tavsiflanadi. Inson hayotining keyingi davrini boshdan kechirar ekan, bir vaqtning o‘zida tarbiyalanuvchi va tarbiyachi bo‘lib, shundan eng muhimi sifatida oila tushunchasi shakllanadi. 19 SOLUTION OF SOCIAL PROBLEMS IN MANAGEMENT AND ECONOMY International scientific-online conference Shu munosabat bilan zamonaviy ijtimoiy-pedagogik makonning dolzarb talabi bo‘lgan yoshlarni oilaviy hayotga tizimli, tizimli, maqsadli psixologik-pedagogik tayyorlash muammosi dolzarb bo‘lib, buni hal etishda katta rol o‘ynashi yaqqol ko‘rinib turibdi. muammo yangi pedagogik kadrlarga tegishli. Oilaning o‘ziga xos ma’naviy-ruhiy psixologik muhiti mehr-muhabbat, g‘amxo‘rlik, hurmat, sezgirlik tufayli boshqa guruhlarga nisbatan yoshlarni oilaviy hayotga tayyorlashda katta afzalliklarga ega. Oiladan tashqarida o‘sgan bolalarda hissiy va aqliy rivojlanish darajasi past. Ular o‘z yaqinlarini sevish qobiliyatini, hamdardlik va hamdardlik qobiliyatini inhibe qildilar. Oilada odamlar o‘rtasida bo‘lishi mumkin bo‘lgan eng yaqin va eng yaqin munosabatlar mavjud bo‘lganligi sababli, ijtimoiy meros qonuni kuchga kiradi. Shu munosabat bilan xalq: “Olma daraxtdan uzoqqa tushmaydi”, deyishadi. Bolalar o‘zlarining fe’l-atvori, fe’l-atvori, xatti-harakatlari bilan ko‘p jihatdan ota-onalariga o‘xshashdir. Har bir oila o‘z muhitini, o‘ziga xos madaniy muhitini rivojlantiradi va u bolaga eng katta ta’sir ko‘rsatadi. Shu munosabat bilan zamonaviy ijtimoiy-pedagogik makonning dolzarb talabi bo‘lgan yoshlarni oilaviy hayotga tizimli, tizimli, maqsadli psixologik-pedagogik tayyorlash muammosi dolzarb bo‘lib, buni hal etishda katta rol o‘ynashi yaqqol ko‘rinib turibdi. muammo yangi pedagogik kadrlarga tegishli. Ammo turli oilalar yosh avlodni oilaviy hayotga tayyorlash uchun katta yoki kamroq imkoniyatlarga ega. Ba’zi ota-onalar o‘z farzandlarini qanday qilib tarbiyalash va rivojlanishiga yordam berishni bilishmaydi, boshqalari buni xohlamaydilar, boshqalari esa har qanday hayotiy sharoit (og‘ir) tufayli buni qila olmaydi. Bundan tashqari, hozirgi vaqtda bir qator sabablarga ko‘ra (nikoh-oila munosabatlaridagi noqulaylik; to‘liq bo‘lmagan oilalarning ko‘pligi; ko‘plab ota- onalarning oilada jinsiy tarbiyani amalga oshirishga tayyor emasligi, ularning oiladagi rolini noto‘g‘ri tushunish) bo‘lajak oila boshlig‘ini tarbiyalash va boshqalar ) oila har doim ham yosh avlodni oilaviy hayotga tayyorlash uchun barcha shart-sharoitlarni ta’minlamaydi. Shunday ekan, oilaviy tarbiyaning ajralmasligiga qaramay, yosh avlodni oilaviy hayotga yanada samarali tayyorlash uchun pedagogik jihatdan barkamol, professional yordam talab etiladi. Aytish joizki, oila haqidagi bilimlarni shakllantirishda ommaviy axborot vositalarining ta’siri katta. Televideniya va kino, matbuot va matbuot ommaviy san’atning eng yaxshi namunalariga ta’sir o‘tkazmay, yosh avlodga tuzatib bo‘lmas zarar yetkazmoqda. “Ommaviy madaniyat va ommaviy axborot vositalarining oilaviy nikohning jamoatchilik fikridagi imidjiga ta’sirining sotsiologik tahlili” loyihasi rahbari A.Antonovning fikricha, “... oila gazeta sahifalari va boshqa ommaviy axborot vositalari materiallaridan. turli muammolarga duchor bo‘lgan, barcha paydo bo‘lgan nizolarni tajovuzkorlik va 20 SOLUTION OF SOCIAL PROBLEMS IN MANAGEMENT AND ECONOMY International scientific-online conference zo‘ravonlik yordamida hal qilishga intilayotgan begona shaxslar sifatida namoyon bo‘ladi", "...badiiy adabiyotda ommaviy ongning oilaviy "oilaviy madaniyatdan oiladan tashqariga o‘tishi. zo ravonlik yordamida hal qilishga intilayotgan begona shaxslar sifatida namoyon bo‘ladi", "...badiiy adabiyotda ommaviy ongning oilaviy "oilaviy madaniyatdan oiladan tashqariga o‘tishi. Talabalarda familistik kompetentlikni oshirish zamirida yosh avlodni oilaviy hayotga tayyorlashda tizimning faoliyati o‘qituvchilar va talabalar jamoasida amalga oshirilishi kerak, shu jumladan ularning tenglik hamkorligi va pedagogik rahbarlikni talabalarning faolligi, havaskorligi va tashabbuskorligi rivojlanishi bilan uyg‘unlashtirish, faollikni faollashtirish. ularning o‘z-o‘zini tarbiyalashi. Agar oilaviy urf-odatlarni uzatishda davomiylik qandaydir qimmatga ega bo‘lsa, G‘arb pop madaniyati tomonidan singdirilgan jinsiy aloqa, zo‘ravonlik va qo‘rquv "ideallari" yosh avlodga tuzatib bo‘lmaydigan zarar etkazadi. Bundan tashqari, na maktab, na universitet dasturlari talabalarni kelajakdagi nikoh va ota-onalik funksiyalariga tayyorlashni nazarda tutmaydi. Talabalar va talabalar o‘zlarining muhim fuqarolik va ijtimoiy burchlarini bajarishga, oila qurish va kelajak farzandlarni tarbiyalashga, katta shaxsiy baxtga erishishga deyarli tayyor emaslar. Bu sohadagi bilimsizlik tufayli yoshlar turmush qurish jarayonida juda ko‘p muammo bor. Odatda ular katta hayotiy muvaffaqiyatsizliklar, ba’zan esa halokatli oqibatlarga olib keladigan dramalar va fojialar sifatida boshdan kechiriladi. Talabalarda familistik kompetentlikni oshirish zamirida yosh avlodni oilaviy hayotga tayyorlashda tizimning faoliyati o‘qituvchilar va talabalar jamoasida amalga oshirilishi kerak, shu jumladan ularning tenglik hamkorligi va pedagogik rahbarlikni talabalarning faolligi, havaskorligi va tashabbuskorligi rivojlanishi bilan uyg‘unlashtirish, faollikni faollashtirish. ularning o‘z-o‘zini tarbiyalashi. Agar oilaviy urf-odatlarni uzatishda davomiylik qandaydir qimmatga ega bo‘lsa, G‘arb pop madaniyati tomonidan singdirilgan jinsiy aloqa, zo‘ravonlik va qo‘rquv "ideallari" yosh avlodga tuzatib bo‘lmaydigan zarar etkazadi. Bundan tashqari, na maktab, na universitet dasturlari talabalarni kelajakdagi nikoh va ota-onalik funksiyalariga tayyorlashni nazarda tutmaydi. Talabalar va talabalar o‘zlarining muhim fuqarolik va ijtimoiy burchlarini bajarishga, oila qurish va kelajak farzandlarni tarbiyalashga, katta shaxsiy baxtga erishishga deyarli tayyor emaslar. Bu sohadagi bilimsizlik tufayli yoshlar turmush qurish jarayonida juda ko‘p muammo bor. Odatda ular katta hayotiy muvaffaqiyatsizliklar, ba’zan esa halokatli oqibatlarga olib keladigan dramalar va fojialar sifatida boshdan kechiriladi. Adabiyotlar: 1. Davletshin M.G., Shoumarov G ‘.B. Zamonaviy o‘zbek oilasi va uning psixologik xususiyatlari O‘zbek oilasining etnopsixologik muammolari: Respublika ilmiy- amaliy anjumani ma’ruzalari qisqacha bayoni. Toshkent, 1993. 2. Oila: ijtimoiy himoya omillari. llmiy-ommabop maqolalar to‘pami., Prof. V. M. Karimova tahriri ostida. Т.: 2007. . 3. R.Mavlonova, O.To‘rayeva, K.Xoliqberdiyev. Pedagogika. « O‘qituvchi». 2008-y. 4. P.A.Mavlonova, B.Normurodova, N.Rahmonqulova. Tarbiyaviy ishlar metodikasi. Т.: «Fan», 2007. 5. J.YoMdoshev va F.YoMdosheva, G.YoMdosheva. Interfaol ta’lim sifat kafolati. Т., 2009. 5. J.YoMdoshev va F.YoMdosheva, G.YoMdosheva. Interfaol ta’lim sifat kafolati. Т., 2009. M.Inomova. Oilada bolalaming ma’naviy-axloqiy tarbiyasi. Т., 1999-y. 7. R.Mavlonova, N.Raxmonqulova. Boshlang'ich ta’limning integratsiyalashgan pedagogikasi. Т., 2009-y. 25. R.Mavlonova, N.Raxmonqulova. Boshlang‘ich ta’lim innovatsiyasi. Т., 2007-y. 7. R.Mavlonova, N.Raxmonqulova. Boshlang'ich ta’limning integratsiyalashgan pedagogikasi. Т., 2009-y. 25. R.Mavlonova, N.Raxmonqulova. Boshlang‘ich ta’lim innovatsiyasi. Т., 2007-y.
https://openalex.org/W2989788958	https://link.springer.com/content/pdf/10.1007/s40097-019-00322-z.pdf	English	null	Comparison of the conductive properties of polyester/viscose fabric treated with Cu nanoparticle and MWCNTs	Journal of nanostructure in chemistry	2,019	cc-by	7,850	Journal of Nanostructure in Chemistry (2019) 9:335–348 https://doi.org/10.1007/s40097-019-00322-z Journal of Nanostructure in Chemistry (2019) 9:335–348 https://doi.org/10.1007/s40097-019-00322-z ORIGINAL RESEARCH Abstract In this work, the specimen of the fabrics (polyester/viscose blend) was prepared. At first, the samples were placed under microwave radiation at different times, and then the optimum condition of treated fabrics (8 min) was selected for treatment. The physical properties and surface morphology of Cu nanoparticle and multi-wall carbon nanotubes (MWCNTs) with differ- ent percentages were measured using dispersing agent, washing performance, stability, and physical properties of the fabric. The image of surface morphology’s specimens was also photographed by scanning electron microscopy (SEM). Afterwards, we measured the specimens’ electrical conductivity properties, according to AATCC 2005-76 standards, and subsequently, K/S, %R, and Lab value of specimens was analyzed using reflection spectrophotometer. In fact, the results indicated that optimum electrical resistivity, which was also the aim of the study, is 9% one weight of fabric (o.w.f.) nanoparticles on the fabric and that electrical resistivity for the values of 9% o.w.f. for CNT is slightly greater than Cu. Keywords Polyester/viscose · Multi-wall carbon nanotubes · Cu nanoparticles · Conductivity Comparison of the conductive properties of polyester/viscose fabric treated with Cu nanoparticle and MWCNTs H. Akbarpour1 · A. Rashidi2 · M. Mirjalili1 · A. Nazari3 Received: 2 September 2019 / Accepted: 9 November 2019 / Published online: 30 November 2019 © The Author(s) 2019 * M. Mirjalili Dr.mirjalili@iauyazd.ac.ir H. Akbarpour Hamid.akbarpour@iauyazd.ac.ir A. Rashidi Rashidi50@yahoo.com A. Nazari a.nazari@iauyazd.ac.ir 1 Department of Textile and Polymer Engineering, Yazd Branch, Islamic Azad University, Yazd, Iran 2 Department of Textile Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran 3 Department of Art and Architect, Yazd Branch, Islamic Azad University, Yazd, Iran Introduction However, it has been found that converging the electronics and textiles, that is e-textiles, Vo Vo (0123 1 3456789) 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 336 336 may be suitable to design the smart materials with the abilities of the performance various functions that are now observed in the non-flexible and rigid electronic products [6]. Electrical resistance of a material indicates how strongly the material opposes the flow of electric current through it. Based on the electrical resistance of the materials, conduc- tive textiles can be classified into the following groups. (1) Insulators: these textile materials have electrical resistance greater than 1011 ohms. They do not allow the electrical current to flow through them. (2) Dissipaters: these textile materials have electrical resistance in the range of 104–1011 ohms. These textile materials do not allow accumulation of electrical charge on their surface. The dissipation increases as the moisture level in the textile increases. (3) Conductors: these are textiles materials with a resistance less than 104 ohms. Such textile materials allow smooth passage of elec- tric current [11]. However, much attention has been paid to the continuous fibers and or yarns consisting of carbon nano- tubes (CNTs) because of the respective inherent capabilities for forming diverse macroscopic objects via a simple knit- ting and or weaving of the yarns or fibers [12]. As Bunshi Fugetsu et al. [13] discovered, the CNT-based dyestuffs can be procured via dispersion of the aggregates of the multiwall CNTs in water by blending the zwitterionic surfactants with the anionic surfactants. Then, we used a dye-printing proce- dure and immediately utilized CNTs to the polyester multi- filament yarns for forming an electrically conductive layer on all filaments of the multi-filament yarn. Actually, the yarns, which have electrical resistivity in the range between 103 and 109 O/cm, have been achieved. Notably, the ones with 103 O/cm resistivity might be applied for forming the soft, flat, and portable electrical heaters via vertical weaving the yarns into the fabrics. In addition, the yarns with 105 O/ cm resistivity might be utilized for antistatic clothing, and the 109 O/cm yarns for brushes for photo-copying machines. Also, Azam Ali et al. [14] investigated the effects of the silver nitrate concentrations on the electrical conductivity. Moreover, 2 g/200 mL provided reasonable levels of the electrical conductivity values. Introduction Additionally, numerous num- bers of dips (i.e., 50–150) in silver nitrate solution have been used to control the amounts of the silver particles deposi- tion. Based on the findings, the greater numbers of the dips generated dense network of the silver particles. Therefore, they led to the increased electrical conductivity. We then dealt with the analysis of the conductive fabrics useful- ness for electro-magnetic shielding capability by coaxially transmitting line technique over a frequency range between 30 MHz and 1.5 GHz. In another study, done by Ebrahim Beygi Chime and et al. [15], using carbon nanotubes and titanium dioxide, it was discovered that the electrical con- ductivity of polyethylene terephthalate (PET), due to the presence of carbon black (CBS) on the surface of the PET fabric increases slightly In this paper it was also indicated that the distribution of nanoparticles of titanium dioxide and nano-carbon black with the use of sodium hypophosphite acid and citric acid works and improves the conductivity of the PET fabric. In this research, unlike other studies done on various fabrics, fibers and resources, measurement of electrical conductivity in a polyester/viscose fabric with nanoparticles was not done and microwave treatment were not performed. This matter will be referred to in this article. Materials and methods In this study, polyester/viscose fabric, which is utilized to manufacture military costumes, with a ratio of 65% polyester and 35% viscose, a count of 32/2 (Ne) warp and 32/1(Ne) weft and with a repeat of 2/1z was used. Multi-wall carbon nanotubes and Cu nanoparticles were purchased from Sigma Aldrich. Acetic acid from Merck, a German Company and the dispersant material Ekalin F from Sandoz (Holzkirchen: Germany) were used in this study. Microwave test The samples were placed in a household microwave oven (LG Electronics CG-2872BC 230 V, 50 Hz MAX.2400 W manufactured by LG Electronics Inc.) for 4, 8 and 12 min (back and front of the fabric) at 900 watts under microwave radiation, to absorb more nanoparticles and create pores on the surface images of scanning electron microscope (SEM) showed that by increasing the duration to 8 min, the surface turns porous and creates a sponge-like condition, which by increasing the duration to 12 min, causes the destruction of the fibers (Fig. 1). Alkaline treatment The fabrics were washed in a batch consisting of 10% o.w.f. Na2S2O4 at 70 °C and10% o.w.f. NaOH for 30 min. Introduction textiles would enable fabricating the comfortable, flexible, and wearable conductive textiles with multi-functional fea- tures [1]. Actually, nano-technology is one of the novel areas with the expectation of having extensive applications in each science and technology like material sciences, mechanics, processing technology of the materials, optics, electron- ics, medication, aero-space, plastics, energy, and textiles. Although, this technology is still in its childhood, research proved that it could be a beneficial instrument for improve- ment of the textiles functions so that it has been attended widely. However, new utilization of the nanotechnology in the textiles would afford diverse features with the potentials for better and novel uses in the products [4, 5]. A number of arts are existing for producing conductive fabrics which include coating metals, conducting polymers over the fabric surfaces, metallic salts films [6], and carbon derivatives [7] or polymeric film synthesis [8–10]. Based on a number of studies, the electrical conductive tex- tiles are demanded both for sensing utilizations and wearable electronics and smart textile applications [1–3], transfer- ring data, monitoring, biomedical and medicine reduction, RF shielding, tissue engineering, super capacitor, battery, fuel cell, transistor, solar cell, organic light emitting diodes (OLEDS) and electro-chromic utilizations. In fact, incorpo- rating the electrically conductive substances into the joint * M. Mirjalili Dr.mirjalili@iauyazd.ac.ir H. Akbarpour Hamid.akbarpour@iauyazd.ac.ir A. Rashidi Rashidi50@yahoo.com A. Nazari a.nazari@iauyazd.ac.ir 1 Department of Textile and Polymer Engineering, Yazd Branch, Islamic Azad University, Yazd, Iran 2 Department of Textile Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran 3 Department of Art and Architect, Yazd Branch, Islamic Azad University, Yazd, Iran * M. Mirjalili Dr.mirjalili@iauyazd.ac.ir H. Akbarpour Hamid.akbarpour@iauyazd.ac.ir A. Rashidi Rashidi50@yahoo.com A. Nazari a.nazari@iauyazd.ac.ir 1 Department of Textile and Polymer Engineering, Yazd Branch, Islamic Azad University, Yazd, Iran 2 Department of Textile Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran 3 Department of Art and Architect, Yazd Branch, Islamic Azad University, Yazd, Iran i According to the studies, the phrase “smart textiles” represents extensive field of studies and products, which expand practicality and utility of the usual fabrics. In fact, textile materials such as the filaments and fibers, which yarn together with the knitted, woven or non-woven structures and can have interaction with the surrounding/users have been called the smart textiles. (1) The fabrics have been washed in a bath consisting of 3% o.w.f. NaOH and 5% o.w.f. non-ionic detergent at 60 °C for 30 min. Then the washing process was continued for 5 cycles. The fabrics have been washed in a bath consisting of 3% o.w.f. NaOH and 5% o.w.f. non-ionic detergent at 60 °C for 30 min. Then the washing process was continued for 5 cycles. Here, W1 refers to the sample weight (g) following the saturation at the standard humidity. Moreover, W2 represents the sample weight (g) dried to the constant weight. BET test The Brunauer–Emmett–Teller method (BET) has a wide uti- lization in the materials science to compute the solids sur- face areas through the physical absorption of gas molecules. Nitrogen adsorption (Micromeritics Gemini III 2375, USA) has been used to evaluate the BET surface area. This test was also carried out for radiation to prove the previous stage. 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 337 Fig. 1 Image of SEM samples of the fabric under microwave treatment in: a raw (2500×), b 4 min (2500×), c 8 min (2500×), d 12 min (2500×) Fig. 1 Image of SEM samples of the fabric under microwave treatment in: a raw (2500×), b 4 min (2500×), c 8 min (2500×), d 12 min (2500×) Fig. 1 Image of SEM samples of the fabric under microwave treatment in: a raw (2500×), b 4 min (2500×), c 8 m es of the fabric under microwave treatment in: a raw (2500×), b 4 min (2500×), c 8 min (2500×), d 12 min (2500×) Treatment with nanoparticles Based on the ASTM D1388-96, Drapometer M003B (SDL) has been used to determine the fabrics bending length in the weft and warp directions. It should be noted that the fabrics tensile strength has been evaluated after conditioning the specimen at 20 °C for 24 h, with 65% relative humidity via an Instron TE-500 from Farayab through a 20-cm scale and across-head speed of 25 cm/min (ASTM D2256). In this stage, we procured the mixes consisting of differ- ent contents of nanoparticles (3, 5, 7 and 9% one weight of fabric (o.w.f.)), 10% o.w.f. dispersing agent, and 2% o.w.f. acetic acid. Afterwards, we added the fabrics into the mixes and inserted them in the bath of the dyeing machine. It is notable that the temperature, pressure, and duration, respec- tively, have been adjusted at 100 °C, 7 bar, and 90 min. Of course, the liquor ratio (L:R) is 50:1 and Ekalin F from Sandoz Company (Holzkirchen: Germany) was used as the dispersing agent. Determining the moisture regain Equation 1 has been used to calculate the moisture regain based on the ASTM procedure 2654-76: Colorimetry analysis (a) The room-temperature conductivity of the samples has been gauged based on the standard four-probe proce- dure through a MCP-HT450 conductivity meter (Dia- instruments Co., Ltd.). According to the research design, a Gretagmacbeth COLO- REYE 7000A spectro-photometer integrated with a com- puter has been used to determine the CIELAB color coordi- nates (L, a, & b) of the samples. Then, we computed the CIELAB color coordinates (L, a, & b) from the reflec- tance outputs for illuminant D65 and 10° observer. (b) A digital multimeter has been used to determine the electrical conductivity of the composite of the dried fabrics has been specified at the room temperature (25 °C). Then, an electrical circuit manufactured by a Hewlett Packard 6634B System DC Power Supply and a digital Hewlett Packard 34401A Multimeter have been used to record the electrical measurements. Washing procedure (1) Moisture regain = {[(W1 −W2 )]∕W2 } × 100. (1) Moisture regain = {[(W1 −W2 )]∕W2 } × 100. Design of experimental This increase in recycled moisture caused a plasticization phenomenon to occur, and increased the length of the rupture. by absorption nanoparticles, are due to the further degra- dation viscose fibers in comparison with polyester fibers. As shown in Table 1, it is clear that BET values increased with increasing the radiation time. This increase from 8 min of radiation to 12 min caused decline in the surface area, indicating destruction of the fibers and them getting stuck together. On the other hand, according to the values given in Table 2, it is clear that the strength of the fabrics dimin- ished by irradiation. The magnitude of the increase in length and the degree of rupture after irradiation increased until 8 min of irradiation and after that decreased. The reduction of strength can be due to the formation of pores on the fib- ers surfaces. Creation of pores also increased absorption of moisture in the sample. This increase in recycled moisture caused a plasticization phenomenon to occur, and increased the length of the rupture. by absorption nanoparticles, are due to the further degra- dation viscose fibers in comparison with polyester fibers. As shown in Table 1, it is clear that BET values increased with increasing the radiation time. This increase from 8 min of radiation to 12 min caused decline in the surface area, indicating destruction of the fibers and them getting stuck together. On the other hand, according to the values given in Table 2, it is clear that the strength of the fabrics dimin- ished by irradiation. The magnitude of the increase in length and the degree of rupture after irradiation increased until 8 min of irradiation and after that decreased. The reduction of strength can be due to the formation of pores on the fib- ers surfaces. Creation of pores also increased absorption of moisture in the sample. This increase in recycled moisture caused a plasticization phenomenon to occur, and increased the length of the rupture. (2) Y = b0 + ∑ biXi + ∑ bijXiXj + ∑ ciX2 i i ≥j i, j = 1, 2, 3 (2) Here, b0 represents an independent term based on the mean value of the experimental plan. bi refers to the regres- sion coefficients, which explain the variables impacts in their linear form. bij stands for the regression coefficients of the interaction terms between the variables. Design of experimental ci represents the coefficients of the variables quadratic form. Images of the SEM fabric containing nanoparticles (Figs. 2, 3) show that for both nanotubes of CNTs and Cu, increasing the amount of nanoparticles to 7 and 9% by weight resulted in the appearance of some NPs on the fiber surface, the presence of which is a lot more at the 9%. In the less than 7% amounts used, the nanoparticles were predomi- nantly contained within the fiber. Particles on the surface usually decrease in their final washings (Figs. 2, 3). The results of strength measurement for CNTS, also showed that the nanoparticles used at the 7% scale were at best, whereas for Cu, with the increase of nanoparticles, the strength fell gradually (Table 3) which is in line with the work done by other scholars [30]. Also, the proposed mechanism for MWCNT and Cu nanoarticles on P/V Fiber indicated to Schemes 1 and 2. FTIR spectroscopy We used FTIR spectroscopy (Thermo Nicolet NEXUS 870 FTIR from Nicolet Instrument Corp., USA) to examine the samples FTIR spectra. SEM study Notably, a SEM (LEO1455VP), made in England, was used in order to investigate the surface morphology of the fiber. (c) Electro-chemical experiments have been performed on the CHI430a electro-chemical work station (USA) with a traditional 3-electrode system. 1 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 338 Table 1 BET test values for raw and microwave-operated samples Vm is the monolayer adsorbed gas quantity and c is the BET constant Time of radiation (min) 0 4 8 12 BET 4.11 4.28 4.86 2.89 Vm 0.506 0.521 0.582 0.355 C 21.526 35.365 29.748 42.752 Design of experimental Vm is the monolayer adsorbed gas quantity and c is the BET constant We applied the central composite design (CCD) for the experimental program with two variables [29]. In fact, the variables involved the amount of irradiation time and percentage of nanoparticles. Results showed the variables range: irradiation time (4.8–12 min) and percentage of nanoparticles (3.1–8.9%), by means of the trial version of the Design Expert 8.0.1.0 software from the Stat-Ease, Inc. (USA). Table 2 Measurement strength and moisture regain test of microwave specimens Table 2 Measurement strength and moisture regain test of microwave specimens Time of radiation (min) Raw 4 8 12 Force at break (kg f) 22.13 20.32 16.11 11.54 Elongation at break (kg f) 7.78 7.97 8.75 7.83 Bending length—warp (cm) 5.84 5.18 4.88 5.36 Bending length—weft (cm) 5.24 5.02 4.57 5.17 Moisture regain (%) 7.21 7.85 8.23 7.05 Tables 7 and 8 (runs 1–8) report designing the polyester/ viscose fabric samples with the irradiation time and per- centage of NPs in detail. Moreover, the variables influences on the outputs Y1 (electrical resistivity) and Y2 (surface resistance) have been tuned by the third-order poly-nominal function below (Eq. 2). It should be noted that although the specimens were selected to three series of time under the radiation but the softness of analyze the range of the changes was selected randomly, in the range of 4–12 min of radia- tion, and showed in eight different runs. On the other hand, laboratory experiments were carried out on the samples with four repeats. by absorption nanoparticles, are due to the further degra- dation viscose fibers in comparison with polyester fibers. As shown in Table 1, it is clear that BET values increased with increasing the radiation time. This increase from 8 min of radiation to 12 min caused decline in the surface area, indicating destruction of the fibers and them getting stuck together. On the other hand, according to the values given in Table 2, it is clear that the strength of the fabrics dimin- ished by irradiation. The magnitude of the increase in length and the degree of rupture after irradiation increased until 8 min of irradiation and after that decreased. The reduction of strength can be due to the formation of pores on the fib- ers surfaces. Creation of pores also increased absorption of moisture in the sample. Results and discussion Before the samples were treated with nanoparticles, they first were operated on under microwave irradiation [16] for different durations. The optimal mode for treating the sur- face of the fabric with nanoparticles was selected, which was found to be 8-min irradiation duration [based on the results of SEM images (Fig. 1)]. It should be noted that the highest percentage of degradation was in the viscose fiber, and the polyester fibers had a better resistance to microwave waves, which indicates that the results of the microscopic projection and the amount of nanoparticles remaining on the fiber surface after the operation of the washing, followed 1 3 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 339 Fig. 2 Image of SEM samples of the fabric under 8-min microwave treatment in: a CNT 3% o.w.f. (2000×), b CNT 5% o.w.f. (5000×), c CNT 7% o.w.f. (5000×), d CNT 9% o.w.f (500×) Fig. 2 Image of SEM samples of the fabric under 8-min microwave treatment in: a CNT 3% o.w.f. (2000×), b CNT 5% o.w.f. (5000×), c CNT 7% o.w.f. (5000×), d CNT 9% o.w.f (500×) Fig. 2 Image of SEM samples of the fabric under 8-min microwave treatment in: a CNT 3% o.w.f. (2000×), b CNT 5% o.w.f. (5000×), c CNT 7% o.w.f. (5000×), d CNT 9% o.w.f (500×) Fig. 3 Image of SEM samples of the fabric under 8-min microwave treatment in: a Cu 3% o.w.f (5000×), b Cu 5% o.w.f (2000×), c Cu 7% o.w.f (5000×), d Cu 9% o.w.f (5000×) Fig. 3 Image of SEM samples of the fabric under 8-min microwave treatment in: a Cu 3% o.w.f (5000×), b Cu 5% o.w.f (2000×), c Cu 7% o.w.f (5000×), d Cu 9% o.w.f (5000×) Nanoparticles value (%) 3 5 7 9 Typical of nanoparticle CNT Cu CNT Cu CNT Cu CNT Cu Force at break (kg f) 22.63 18.41 23.15 15.11 24.28 11.25 12.02 7.37 Elongation at break (%) 7.12 7.97 6.98 8.28 6.56 8.78 7.67 9.16 Bending length—warp (cm) 6.11 5.32 6.58 5.11 6.01 4.89 5.12 4.28 Bending length—weft (cm) 5.98 5.18 6.25 5.06 5.48 4.62 4.89 4.13 Table 4 indicates the rate of washing stability for fabrics treated with nanoparticles. The results showed an increase in Cu uptake, compared to CNTS in the fabric, and higher fiber penetration of Cu nanoparticles than that of other nanoparti- cles this method has not been observed by others. Table 3 Measurement strength test of microwave specimens in 8 min with different amounts of nanoparticles Results and discussion are given in Table 5. In another way, the conductivity testing of nanoparticles treated with a digital multimeter and a DC voltage source under the AATCC 1995-76 Method at 25 °C was carried out. In this approach, two copper electrodes with a size of (20 × 20 mm2) were separated at a distance of 20 mm, and then, a 1 kg weight was inserted over the fabric sample (30 × 60 mm2). Afterwards, the surface resistance (Rs) was estimated by Eq. 3: Moreover, the conductivity test was carried out in three ways: At first, the electrical resistance of the fabric was measured using a digital multimeter, the results of which 1 3 340 Journal of Nanostructure in Chemistry (2019) 9:335–348 Scheme 1 The proposed mechanism of MWCNT on P/V Fiber Scheme 1 The proposed mechanism of MWCNT on P/V Fiber involving an auxiliary electrode (platinum wire), a refer- ence electrode (Ag/AgCl), and working electrode (modified GCE). We conducted each experiment at room temperature. Then, we applied the cyclic voltammetry (CV) to describe the electrode resistance functions. Afterwards, 5 mM [Fe (CN) 6]3−/4− has been employed as the probe, and 0.1 M KCl has been applied as the supporting electrolyte. The results of the test of the conductivity (flow-to-voltage) of the micro- wave treated samples, for a duration of 8 min, for different voltammetric nanoparticles are shown in Figs. 4 and 5. Fig- ures 4 and 5 show a comparison diagram of the ratio of the voltage intensity to the current of the samples treated with different nanoparticles, compared to the untreated sample with the nanoparticles after microwave operation (8 min.). As shown in the plots, the difference is increased by increas- ing the percentage of nanoparticles. On the other hand, this difference in the samples treated by CNT has a special regu- larly compared to Cu-treated samples, which seems to indi- cate the conductivity quality. (3) RS(Ω∕square) = (W∕D) × R. Here, R represents the resistance gauged by the multi- meter. D and W, respectively, refer to the distance between 2 electrodes and sample width. As shown in Table 5, the best electrical resistance is observed in the amounts of 9% consumable nanoparticles. Furthermore, at 3%, electrical resistance for CNTS dropped, suddenly, however, a similarity was observed at 9% and 5% nanoparticles. Moreover, the resulted quantities indicated an upward trend in conductivity. 1 3 Table 4 Measurement of the stability of the washing of the fabrics treated with nanoparticles Results and discussion For the raw sample, the appeared bands at 3431, 2964, 2340, Scheme 2 The proposed mechanism of MWCNT on P/V Fiber Table 4 Measurement of the stability of the washing of the fabrics treated with nanoparticles Nanoparticles value (%) 3 5 7 9 Typical of nanoparticle CNT Cu CNT Cu CNT Cu CNT Cu (%) Increase of initial weight 2.88 2.93 4.29 4.79 6.12 6.58 7.2 7.39 Increase of weight after wash- ing (%)/(cycle 1-relative to raw fabric) 2.79 2.90 4.1 3.72 5.95 5.44 5.6 5.98 Increase of weight after wash- ing (%)/(cycle 5-relative to raw fabric) 2.61 2.89 3.95 3.99 5.88 6.32 3.7 4.25 Table 5 Electrical resistivity and conductivity of a sample of fabrics under 8 min of microwave irradiation treated with MWCNT and Cu nanoparticles Nanoparticles value (%) 3 5 7 9 Typical of nanoparticle CNT Cu CNT Cu CNT Cu CNT Cu R (Ω) 131251 26677 10368 10352 5369 2672 1610 1753 σ (S/cm) 0.0023 0.0133 0.0247 0.0261 0.0651 0.1338 0.2335 0.3402 Journal of Nanostructure in Chemistry (2019) 9:335–348 341 Journal of Nanostructure in Chemistry (2019) 9:335–348 Scheme 2 The proposed mechanism of MWCNT on P/V Fiber Table 4 Measurement of the stability of the washing of the fabrics treated with nanoparticles Nanoparticles value (%) 3 5 7 9 Typical of nanoparticle CNT Cu CNT Cu CNT Cu CNT Cu (%) Increase of initial weight 2.88 2.93 4.29 4.79 6.12 6.58 7.2 7.39 Increase of weight after wash- ing (%)/(cycle 1-relative to raw fabric) 2.79 2.90 4.1 3.72 5.95 5.44 5.6 5.98 Increase of weight after wash- ing (%)/(cycle 5-relative to raw fabric) 2.61 2.89 3.95 3.99 5.88 6.32 3.7 4.25 Table 5 Electrical resistivity and conductivity of a sample of fabrics under 8 min of microwave irradiation treated with MWCNT and Cu nanoparticles Nanoparticles value (%) 3 5 7 9 Typical of nanoparticle CNT Cu CNT Cu CNT Cu CNT Cu R (Ω) 131251 26677 10368 10352 5369 2672 1610 1753 σ (S/cm) 0.0023 0.0133 0.0247 0.0261 0.0651 0.1338 0.2335 0.3402 Table 5 Electrical resistivity and conductivity of a sample of fabrics under 8 min of microwave irradiation treated with MWCNT and Cu nanoparticles Also, as shown in Table 6, the K/S-values of the samples augmented by increasing the percentage of nanoparticles and decreased the R% values. Results and discussion However, in Cu nanoparticles, there was an increase in conductivity up to 7%, in compari- son with CNTS. On the other hand, the results showed the superiority of surface strength carbon nanotubes compared to Cu nanoparticles. The values of the electrical conductiv- ity obtained are compared with those of other researchers in Table 13 [23–27]. In the third step, the conductivity of fabric treated with nanoparticles was measured using a CHI430A electro-chem- ical work station (USA), which has a three electrode system 341 Journal of Nanostructure in Chemistry (2019) 9:335–348 Also, as shown in Table 6, the K/S-values of the samples augmented by increasing the percentage of nanoparticles and decreased the R% values. However, it can be observed that there are similarities between the numerical values of K/S and R% and samples treated with 9% nanoparticles for MWCNT and 7% nanoparticles for Cu, respectively; this is the equivalent of the coefficient equal to the propagation of samples. On the other hand, with respect to the values in the table, the values of L, C and h° decreased with increasing the nanoparticle contents, and the values of b decreased with increasing the amount of nanoparticles for Cu and for MWCNT, it firstly rose and then fell, steadily. The FTIR spectra of samples are shown in Fig. 6. Table 5 Electrical resistivity and conductivity of a sample of fabrics under 8 min of microwave irradiation treated with MWCNT and Cu nanoparticles Results and discussion However, it can be observed that there are similarities between the numerical values of K/S and R% and samples treated with 9% nanoparticles for MWCNT and 7% nanoparticles for Cu, respectively; this is the equivalent of the coefficient equal to the propagation of samples. On the other hand, with respect to the values in the table, the values of L, C and h° decreased with increasing the nanoparticle contents, and the values of b decreased with increasing the amount of nanoparticles for Cu and for MWCNT, it firstly rose and then fell, steadily. i The FTIR spectra of samples are shown in Fig. 6. For the raw sample, the appeared bands at 3431, 2964, 2340, 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 342 y Fig. 4 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a untreated, b 3% Cu, c 5% Cu, d 7% Cu, e 9% Cu Fig. 4 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a untreated, b 3% Cu, c 5% Cu, d 7% Cu, e 9% Cu the voltages of the samples exposed to microwave radiation for 8 min for: a untreated, b 3% Cu, c 5% Cu, d 7% Cu Fig. 4 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a untreated, b 3% Cu, c 5% Cu, d 7% Cu, e 9% Cu 1715–1736, 1408, 1233,1000–1100 and 690–900 cm−1 were attributed to OH (H-bonded), aldehyde, carboxylic acids, C=O, C=C (aromatic), C–H bending, C–O (esters or ethers) and aromatics (out of plane bend), respectively (Fig. 6a); whereas, in the sample treated with MWCNTs the bands observed at 3446, 2913, 1640, 1000–1100 and 690–900 cm−1 related to OH (H-bonded), aldehyde, C=C (aromatic), C–O (esters or ethers) and aromatics (out of plane bend), respectively (Fig. 6b). Moreover, in Fig. 6c, the sample treated with Cu nanoparticles indicated approx- imately the same results. As shown in Figs. 7 and 8, the decrease in the peak intensity of carboxylic acid groups (2362 cm−1) is caused by the existence of the Cu NPs and MWCNTs onto the fabrics. On the other hand, the peak appearing in 1417 cm−1 related to MWCNTs [28] and the peaks appearing 592 cm−1 and 500 cm−1 attributed to Cu nanoparticles [29]. 1 3 Statistical analyses Based on the analyses, optimizing the use of the NPs would suggest the fundamental contribution to the outperformance Journal of Nanostructure in Chemistry (2019) 9:335–348 343 Fig. 5 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a 3% CNT, b 5% CNT, c 7% CNT, d 9% CNT Fig. 5 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a 3% CNT, b 5% CNT, c 7% CNT, d 9% CNT Fig. 5 Flow rate diagram of the voltages of the samples exposed to microwave radiation for 8 min for: a 3% CNT CNT Table 6 K/S, %R, CIELab and CIEch values of samples of microwave-irradiated fabrics for 8 min under treatment of MWCNT and Cu nanoparticles Nanoparticles value (%) Raw 3 5 7 9 K/S (λmax = 400 nm) MWCNT 0.068 4.9 5.94 9.916 10.9 Cu 6.9 7.93 11.05 12.14 %R (λmax = 700 nm) MWCNT 84.34 14.87 9.88 6.87 5.84 Cu 16.71 11.73 6.88 5.82 L* MWCNT 92.49 44.82 36.82 30.33 29.21 Cu 38.51 30.53 27.34 25.26 a* MWCNT − 0.32 3.12 2.87 2.02 2.01 Cu 2.59 2.59 2.42 2.31 b* MWCNT 4.12 7.52 6.88 5.35 5.42 Cu 3.25 3.08 2.95 2.91 C MWCNT 4.13 6.69 5.25 4.86 4.67 Cu 6.99 4.19 3.82 3.49 h° MWCNT 94.45 74.17 69.91 61.36 57.62 Cu 64.29 52.69 50.65 47.57 Table 6 K/S, %R, CIELab and CIEch values of samples of microwave-irradiated fabrics for 8 min under treatment of MWCNT and Cu nanoparticles 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 344 Fig. 6 ATR-FTIR analyses of polyester/viscose blend fabric: a raw sample, b sample of microwave-irradiated fabrics for 8 min under treatment of MWCNT, c sample of microwave-irradiated fabrics for 8 min under treatment of Cu nanoparticles widely employed in different biochemical, nano-chemi- cal, and chemical procedures for analyzing the effects of the independent variables and optimizing the procedure responses via the suitable values of the parameters [18–22]. The present research was carried out on the basis of the CCD and RSM. Eight experimental CCD designed runs have been accomplished, as demonstrated by Tables 7 and 8. As seen, the impacts of the independent variables like the irradiation time and nanoparticles percentage on the response surface have been evaluated. This justified the electrical resistivity and conductivity properties of treated fabric samples. Statistical analyses Final equation in terms of actual factors related to elec- trical resistivity and conductivity of the sample of fabrics treated with Cu nanoparticles and MWCNTS are presented in Eqs. (4), (5), (6) and (7), respectively: (4) R(Ω) = +96540.82306 −6820.09154 × irradiation time −12668.11108 × Cu nanoparticle + 945.36356 × irradiation time × Cu nanoparticle, (4) (5) 휎(S∕cm) = −0.681510 + 0.054816 × irradiation time + 0.152864 × Cu nanoparticle −0.011227 × irradiation time × Cu nanoparticle, (5 휎(S∕cm) = −0.681510 + 0.054816 × irradiation time + 0.152864 × Cu nanoparticle −0.011227 × irradiation time × Cu nanoparticle, (5) (6) R(Ω) = +6.92 × 105 −49574.332 × irradiation time −1.303 × 105 × CNT nanoparticle + 2927.025 × irradiation time × CNT nanoparticle + 1649.91 × irradiation time2 + 6906.983 × CNT nanoparticle2, (6) (7) 휎(S∕cm) = +0.067 −0.0287 × irradiation time −0.000051 × CNT nanoparticle −0.0061 × irradiation time × CNT nanoparticle + 0.00355 × irradiation time2 + 0.0076 × CNT nanoparticle. Fig. 6 ATR-FTIR analyses of polyester/viscose blend fabric: a raw sample, b sample of microwave-irradiated fabrics for 8 min under treatment of MWCNT, c sample of microwave-irradiated fabrics for 8 min under treatment of Cu nanoparticles (7) The resulting statistical models have been used to draw the response surface (Eqs. 4–7), and then, the relationship between all independent variables as well as electrical resistivity and conductivity of fabric specimens have been achieved. Figures 7 and 8, respectively, depict the response surface of the fabric samples. of the textile fabrics. Here, it should be noted that contrary to the traditional optimization, the statistical optimization procedures and interactions between variables for produc- tion of the procedure response may be considered. In fact, RSM is one of the robust statistical techniques of analyzing several variables as a result of fewer essential experimental trials in comparison with the “one-factor-at-a-time” tech- nique. Moreover, RSM is one of the effective mathematical approaches of optimization of the complicated procedures that are able to produce an empirical pattern for evaluat- ing the correlation between some controlled experimental parameters and the observed outputs. Actually, RSM is of the textile fabrics. Here, it should be noted that contrary to the traditional optimization, the statistical optimization procedures and interactions between variables for produc- tion of the procedure response may be considered. Statistical analyses In fact, RSM is one of the robust statistical techniques of analyzing several variables as a result of fewer essential experimental trials in comparison with the “one-factor-at-a-time” tech- nique. Moreover, RSM is one of the effective mathematical approaches of optimization of the complicated procedures that are able to produce an empirical pattern for evaluat- ing the correlation between some controlled experimental parameters and the observed outputs. Actually, RSM is In the next stage, Design-Expert software, with irradiation time of 8 min for both Cu nanoparticles and CNT has been used to obtain the optimal conditions of the properties of elec- trical resistivity and conductivity of fabric specimens. Then, analysis of variance (ANOVA) has been applied for analyzing the data so that the interactions between independent variables and responses have been established. Afterwards, ANOVA 1 3 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 345 Fig. 7 Response surface for a electrical resistivity and b conductivity as a function of irradiation time and Cu nanoparticles for fabric samples Fig. 7 Response surface for a electrical resistivity and b conductivity as a function of irradiation time and Cu nanoparticles for fabric samples Fig. 8 Response surface for a electrical resistivity and b conductivity as a function of irradiation time and CNT nanoparticles for fabric samples Fig. 8 Response surface for a electrical resistivity and b conductivity as a function of irradiation time and CNT nanoparticles for fabric samples Fig. 8 Response surface for a electrical resistivity and b conductivity as a function of irradiation time and CN 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 346 Table 7 CCD for electrical resistivity and conductivity of sample of fabrics treated with Cu nanoparticles at different irradiation times Run Factor Responses Irradiation time (min) Go-nanopar- ticle (%) R (Ω) σ (S/cm) Control 1 0 6 74652 0.0015 Control 2 8 0 Unlimited Unlimited 1 8 6 6550 0.07943 2 8 6 6512 0.07995 3 8 6 6490 0.07894 4 4.8 4.2 32399 0.0345 5 4.8 7.8 3595 0.3851 6 12 6 9768 0.1199 7 9.2 8.9 1507 0.2925 8 9.2 3.1 23970 0.01195 Table 7 CCD for electrical resistivity and conductivity of sample of fabrics treated with Cu nanoparticles at different irradiation times has been used to analyze the outputs in order to evaluate the electrical resistivity and conductivity of the fabric samples (Tables 9, 10, 11, 12). Statistical analyses As shown, the introduced model of the fabric samples with Cu nanoparticles and MWCNTS for electrical resis- tivity has been statistically significant at F value of 8.56 and 2.070E+06, and prob values greater than F, 0.0325 and < 0.0001, respectively (Tables 9, 11). In addition, the selected model of the treated fabric specimens with Cu nan- oparticles and MWCNTS for conductivity has been signifi- cant at F value of 9.54 and 9623.67 and prob values greater than F, 0.0270 and < 0.0001, respectively (Tables 10, 12). Comparison of electrical conductivity results with previous results Also, Table 13 shows the comparative results of the electri- cal conductivity obtained with the results obtained in previ- ous papers. Table 8 CCD for electrical resistivity and conductivity of sample of fabrics treated with MWCNTS at different irradiation times Conclusion The effects of nanoparticles (MWCNT and Cu) treatment on the conductivity of the polyester/viscose fabric were investi- gated and their benefit demonstrated. Studies using SEM, con- ductivity test, electrical resistance, strength and voltametric charts of the fabric have shown that under optimal microwave pre-radiation conditions, sample treated with MWCNT and Cu nanoparticles exhibited improved conductivity. Statistical results using the central composite design (CCD) methodol- ogy further established that the fabric samples treated with MWCNT nanoparticles had better uniformity and higher con- ductivity compared with that treated with Cu nanoparticles. Conclusion Source Sum of squares df Mean square F value p value Probability > F Model 7.198E+08 3 2.399E+08 8.56 0.0325 Significant A-irradiation time 5.271E+07 1 5.271E+07 1.88 0.2421 B-Cu nanoparticle 5.865E+08 1 5.865E+08 20.93 0.0102 AB 8.047E+07 1 8.047E+07 2.87 0.1654 Residual 1.121E+08 4 2.802E+07 Lack of fit 1.121E+08 2 5.604E+07 60865.70 < 0.0001 Significant Pure error 1842.67 2 921.33 Cor total 8.319E+08 7 1 3 1 3 1 3 Journal of Nanostructure in Chemistry (2019) 9:335–348 347 Table 10 ANOVA outputs of conductivity for the fabric samples with Cu nanoparticles at different irradiation times Source Sum of squares df Mean square F value p value Probability > F Model 0.1071 3 0.0357 9.54 0.0270 Significant A-irradiation time 0.0063 1 0.0063 1.68 0.2643 B-Cu nanoparticle 0.0894 1 0.0894 23.89 0.0081 AB 0.0113 1 0.0113 3.03 0.1566 Residual 0.0150 4 0.0037 Lack of fit 0.0150 2 0.0075 29347.65 < 0.0001 Significant Pure error 5.102E−07 2 2.551E−07 Cor total 0.1221 7 Table 11 ANOVA outputs of electrical resistivity for the fabric samples with MWCNTS at different irradiation times Source Sum of squares df Mean square F value p value Probability > F Model 1.952E+10 5 3.904E+09 2.070E+06 < 0.0001 Significant A-irradiation time 1.260E+09 1 1.260E+09 6.682E+05 < 0.0001 B-Cu nanoparticle 1.301E+10 1 1.301E+10 6.896E+06 < 0.0001 AB 7.714E+08 1 7.714E+08 4.089E+05 < 0.0001 Residual 7.467E+08 1 7.467E+08 3.958E+05 < 0.0001 Lack of fit 4.140E+09 1 4.140E+09 2.195E+06 < 0.0001 Pure error 3772.67 2 1886.33 Cor total 1.952E+10 7 Table 12 ANOVA outputs of conductivity for the fabric samples with MWCNTS at different irradiation times Source Sum of squares df Mean square F value p value Probability > F Model 0.0541 5 0.0108 9623.67 0.0001 Significant A-irradiation time 0.0029 1 0.0029 2586.67 0.0004 B-Cu nanoparticle 0.0404 1 0.0404 35936.37 < 0.0001 AB 0.0033 1 0.0033 2976.53 0.0003 Residual 0.0035 1 0.0035 3072.84 0.0003 Lack of fit 0.0050 1 0.0050 4457.50 0.0002 Pure error 2.247E−06 2 1.123E−06 Cor total 0.0541 7 Table 13 Comparison of results with those reported in previously published Type of goods Optimum sample PV–MWCNT Irradiation time: 9.2 (% 8.9 o.w.f) Optimum sample PV–Cu Irradiation time: 9.2 (% 8.9 o.w.f) Wool–MWCNT (5.5 g/L) [23] Aminolyzed polyester–% 0.2 GO [24] Nylon–nano- silver [25] Polyester– MWCNT (30 g/L) [26] Cotton–GO 1% [27] R(Ω/Sq) 1384 1507 1.07 × 107 0.11 × 106 4.5 103 to 109 55 Table 13 Comparison of results with those reported in previously published Table 11 ANOVA outputs of electrical resistivity for the fabric samples with MWCNTS at different irradiation times Table 12 ANOVA outputs of conductivity for the fabric samples with MWCNTS at different irradiation times References Open Access This article is distributed under the terms of the Crea- tive Commons Attribution 4.0 International License (http://creativeco mmons.org/licenses/by/4.0/), which permits unrestricted use, distribu- tion, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 1. 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https://openalex.org/W2061629872	https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0057377&type=printable	English	null	Is Accessing Dental Care Becoming More Difficult? Evidence from Canada's Middle-Income Population	PloS one	2,013	cc-by	6,283	Introduction or co-payments [1,9,10]. At the same time, the availability of employment-based dental insurance decreased as Canadian firms began to change the nature of employment by increasing part-time and temporary positions, thus limiting the growth of unionization [1]. Importantly, it is full-time, permanent, unionized jobs in large firms that have benefitted the most from the presence of dental insurance, and it is low- and middle-income employees that have observed the greatest impacts to their workplace structures in this way [1,11,12,13]. Even in the United States, employment-based dental insurance among lower-middle income Americans declined from 80.2% in 1999 to 77.4% by 2002 [14]. In Canada, recent data have shown that challenges in accessing dental care no longer just affect those with no or low incomes, but also those above them on the economic ladder [1–5]. In this regard, specific attention has been placed on working poor Canadians, namely those that do not qualify for public dental care programs, yet generally do not have jobs that offer employment- based dental insurance, which is the country’s dominant form of financing dental care for individuals and families [1–5]. It has also been anecdotally reported that in light of the recent global economic downturn, middle-income families are now contacting local public health agencies in an effort to access public dental care programs [1]. Additionally, the costs of and demand for dental care have increased sharply in Canada [15], while incomes have remained nearly stagnant over the past 25 years [16]. When adjusting for inflation, total per capita dental care expenditures rose from approximately $6 in 1960 to close to $50 by 2008 ($360 in current dollars), an increase of approximately 730% [15]. As a measure of demand, the utilization of dental care rose from 49.5%, as reported in the Nutrition Canada National Survey (1970–1972), to 74.5%, as reported in the recent Canadian Health Measures Survey (2007–2009) [15]. At the same time, family earnings stagnated or declined for those at the bottom of the income distribution, while rising substantially at the top [16]. These challenges in accessing dental care are fundamentally linked to issues of affordability. In Canada, income and insurance are known to be the dominant predictors of the utilization of and access to dental care, specifically because of their ability to mitigate the costs of care [2,6,7,8]. Abstract Importantly, it is full-time, permanent, unionized jobs in large firms that have benefitted the most from the presence of dental insurance, and it is low- and middle-income employees that have observed the greatest impacts to their workplace structures in this way [1,11,12,13]. Even in the United States, employment-based dental insurance among lower-middle income Americans declined from 80.2% in 1999 to 77.4% by 2002 [14]. Chantel Ramraj, Laleh Sadeghi, Herenia P. Lawrence, Laura Dempster, Carlos Quin˜ onez Discipline of Dental Public Health, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada Discipline of Dental Public Health, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada February 2013 \| Volume 8 \| Issue 2 \| e57377 Abstract Objective: To explore trends in access to dental care among middle-income Canadians. Methods: A secondary data analysis of six Canadian surveys that collected information on dental insurance coverage, cost- barriers to dental care, and out-of-pocket expenditures for dental care was conducted for select years from 1978 to 2009. Descriptive analyses were used to outline and compare trends among middle-income Canadians with other levels of income as well as national averages. Results: By 2009, middle-income Canadians had the lowest levels of dental insurance coverage (48.7%) compared to all other income groups. They reported the greatest increase in cost-barriers to dental care, from 12.6% in 1996 to 34.1% by 2009. Middle-income Canadians had the largest rise in out-of-pocket expenditures for dental care since 1978. Conclusions: This study suggests that affordability issues in accessing dental care are no longer just a problem for the lowest income groups in Canada, but are now impacting middle-income earners as a consequence of their lack of, or decreased access to, comprehensive dental insurance. Citation: Ramraj C, Sadeghi L, Lawrence HP, Dempster L, Quin˜onez C (2013) Is Accessing Dental Care Becoming More Difficult? Evidence from Canada’s Middle- Income Population. PLoS ONE 8(2): e57377. doi:10.1371/journal.pone.0057377 Citation: Ramraj C, Sadeghi L, Lawrence HP, Dempster L, Quin˜onez C (2013) Is Accessing Dental Care Becoming More Difficult? Evide Income Population. PLoS ONE 8(2): e57377. doi:10.1371/journal.pone.0057377 Editor: Robert A. Burne, University of Florida, United States of America Editor: Robert A. Burne, University of Florida, United States of America Received December 3, 2012; Accepted January 22, 2013; Published February 20, 2013 Copyright: 2013 Ramraj et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: The authors would like to thank the funders of this project, the Population Health Improvement Research Network (PHIRN) of the Applied Health Research Network Initiative (AHRNI), of the Government of Ontario. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. E-mail: chantel.ramraj@utoronto.ca or co-payments [1,9,10]. At the same time, the availability of employment-based dental insurance decreased as Canadian firms began to change the nature of employment by increasing part-time and temporary positions, thus limiting the growth of unionization [1]. Study design and samples This was a secondary data analysis of a series of surveys provided by Statistics Canada: the Canadian Health Measures Survey (CHMS), Canadian Community Health Surveys (CCHS), National Population Health Surveys (NPHS), General Social Surveys (GSS), Surveys of Family Expenditures/Health Expendi- tures (FAMEX/SHS), and Workplace and Employee Surveys (WES). These surveys were accessed online through the University of Toronto’s Data Library, or through Statistics Canada’s Research Data Centre (RDC) at the University of Toronto. Due to the nature of each survey’s sample population, the current study was limited to Canadians aged 12 years of age and older living in private households across the country. The overall period of observation extended from 1978 to 2009. Descriptive analyses were conducted to outline historical trends in self-reported dental insurance coverage, cost-barriers to dental care and out-of-pocket expenditures. These results were stratified by several socio-demographic factors (age, sex, education, employment characteristics, etc.). Comparisons were made between trends for middle-income earners to other income levels (i.e. lowest, lower-middle, higher-middle, and the highest income levels) and to national averages. N The Canadian Health Measure Survey (CHMS) was conduct- ed from March 2007 to February 2009 and collected health measures, including oral health measures, from approximately 5,600 people, representing 97% of the Canadian population between 6 and 79 years of age [15]. N The Canadian Community Health Surveys (CCHS) are a series of cross-sectional surveys collecting information on the health status, health care utilization and health determinants of Canadians. Data are collected from Canadians aged 12 years of age and older; a sample of 65,000 respondents is required on an annual basis representing 98% of the Canadian population [17]. Data for the years 2001, 2003, 2005 and 2007 were used. SPSS 18.0 was used for the analysis of data. Sample weights as calculated by Statistics Canada were applied to all data, where applicable. Data were displayed graphically for the years where information was available nationally. Additionally, indexes of change were calculated using the following formula: the value of each of the years in a time period/beginning year’s value x 100. According to the Organization for Economic Co-operation and Development (OECD), an index of change can help to highlight change in a series from one period to another [19]. N The National Population Health Surveys (NPHS) collect information on the health status and health care utilization of the Canadian population and includes persons from all ages. Self-reported dental insurance coverage p g Figure 1 shows self-reported dental insurance coverage in Canada by income adequacy for select years. As can be seen, the proportion of middle-income Canadians with dental insurance increased from 43.1% (95% CI: 42.4, 43.9) in 1996 to 48.7% (95% CI: 47.4, 50.0) by 2009. Importantly though, by 2009, middle- income earners had the lowest levels of dental insurance coverage when compared to all other income groups. Figure 1 also compares these results to national trends in dental insurance. While the national average rose steadily between 1996 and 2009, from 55.1% (95% CI: 54.7, 55.5) to 68.1% (95% CI: 66.9, 69.3), the proportion of insured middle-income earners declined from 43.1% (95% CI: 42.4, 43.9) in 1996 to 40.4% (95% CI: 39.8, 41.0) by 2003, then increased to 48.7% (95% CI: 47.4, 50.0) by 2009. Figure 2 shows dental insurance coverage levels among middle- income full-time and part-time workers compared to the national averages for these groups. When compared to national averages, middle-income earners had lower levels of dental insurance coverage for both full- and part-time workers. Middle-income full- and part-time workers also had the same level of dental insurance in 2009 (47.6% (95% CI: 46.3, 48.9) vs. 47.5% (95% CI: 46.2, 48.8)), yet nationally, for all incomes, a difference was seen between the two, with a greater percentage of full-time workers Figure 1 shows self-reported dental insurance coverage in Canada by income adequacy for select years. As can be seen, the proportion of middle-income Canadians with dental insurance increased from 43.1% (95% CI: 42.4, 43.9) in 1996 to 48.7% (95% CI: 47.4, 50.0) by 2009. Importantly though, by 2009, middle- income earners had the lowest levels of dental insurance coverage when compared to all other income groups. Figure 1 also compares these results to national trends in dental insurance. While the national average rose steadily between 1996 and 2009, from 55.1% (95% CI: 54.7, 55.5) to 68.1% (95% CI: 66.9, 69.3), the proportion of insured middle-income earners declined from 43.1% (95% CI: 42.4, 43.9) in 1996 to 40.4% (95% CI: 39.8, 41.0) by 2003, then increased to 48.7% (95% CI: 47.4, 50.0) by 2009. N The Surveys of Family Expenditures/Health Expenditures (FAMEX/SHS) record detailed annual spending patterns for a nationally and regionally representative sample of private households in Canada. Data from the FAMEX/SHS from 1978 (N = 9,356 households) to 2009 (N = 16,758 households) were used. Self-reported dental insurance coverage N The Workplace and Employee Surveys (WES) have collected data annually from 1999, encompassing both longitudinal and cross-sectional components. Employer side data of the cross- sectional components from 1999 (N = 6,322) to 2006 (N = 6,312) were used. Figure 2 shows dental insurance coverage levels among middle- income full-time and part-time workers compared to the national averages for these groups. When compared to national averages, middle-income earners had lower levels of dental insurance coverage for both full- and part-time workers. Middle-income full- and part-time workers also had the same level of dental insurance in 2009 (47.6% (95% CI: 46.3, 48.9) vs. 47.5% (95% CI: 46.2, 48.8)), yet nationally, for all incomes, a difference was seen between the two, with a greater percentage of full-time workers Assess to Dental Care for Middle-Income Canadians middle-income group was defined by using ‘income adequacy,’ which takes into account the total household income of a family and the number of people living in that household [15]. This study used Statistics Canada’s definition of middle-income for the years 1994-2009, which includes households with: one or two people earning $15,000–$29,999, three or four people earning $20,000– 39,999 and 5 or more people earning $30,000–$59,999. Since this information was not available prior to 1994, middle-income adequacy was calculated by converting the original income brackets to current dollars for each year prior to 1994 using the Consumer Price Index (CPI) of Canada. The CPI is the most widely used index for household or family incomes, reflecting average spending patterns by consumers in Canada. The annual CPI from 1971 to 2010 is reported by Statistics Canada. The year 2002 was chosen as the base year as it was the median year in the observation period. To convert current dollars of any year to constant dollars, the amount was multiplied by the index of the base year divided by the index of the chosen year [18]. For example, $10,000 in 1997 (CPI of 90.4) would be $11,602 in 2002 dollars ($10,0006100/90.4 = $11,602). These amounts were rounded to the nearest thousand. greater challenges in accessing dental care. As a result, this study explored the issue of access to dental care among middle-income Canadians, by outlining trends in: self-reported dental insurance coverage; self-reported cost-barriers to dental care; and out-of- pocket expenditures for dental care. Study design and samples These studies used files of the cross-sectional components in 1996/1997 (N = 73,402) and 1998/1999 (N = 15,249). Results N The General Social Surveys (GSS) collect information on social support and living conditions over time. Data on dental care were collected in 1985 (N = 11,200) and 1991 (N = 11,924). N The General Social Surveys (GSS) collect information on social support and living conditions over time. Data on dental care were collected in 1985 (N = 11,200) and 1991 (N = 11,924). Introduction Yet evidence has shown that dental insurance plans have changed dramatically over the last 20 years and have been diminished in terms of both their quality and availability [1]. For example, the quality of dental insurance plans decreased following a recession in the 1990s, as Canadian firms found ways to cost-contain in part by altering benefit plans through the limiting of annual maximums and/or services, and/or through the introduction or expansion of deductibles, co-insurance Overall then, with changes to insurance, incomes, the costs of dental care, and a greater demand for it, there is a reasonable case for the anecdotal reports of middle-income Canadians facing February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org 1 February 2013 \| Volume 8 \| Issue 2 \| e57377 Assess to Dental Care for Middle-Income Canadians Assess to Dental Care for Middle-Income Canadians Figure 1. Dental insurance coverage in Canada, by income adequacy and the national average, select years. The proportion of middle-income Canadians with dental insurance increased from 43.1% in 1996 to 48.7% by 2009. By 2009, middle-income earners had the lowest levels of dental insurance coverage when compared to all other income groups. While the national average rose steadily between 1996 and 2009, from 55.1% to 68.1%, the proportion of insured middle- income earners declined from 43.1% in 1996 to 40.4% by 2003, then increased to 48.7% by 2009. doi:10 1371/journal pone 0057377 g001 cost-barriers to the national average, a general increase can be seen in both groups from 1996 to 2009. However, the level of growth is much higher among the middle-income group. While in 1996 the level reporting cost-barriers among middle-income Canadians was only slightly different from national levels (12.6% (95% CI: 11.7, 13.7) vs. 12.2% (95% CI: 11.6, 12.8)), by 2009 middle-income Canadians made such reports about twice more than the national average (34.1% (95% CI: 32.9, 35.3) vs. 17.3% (95% CI: 16.3, 18.3)). Importantly, between 2003 and 2009 when reports made by middle-income Canadians increased from 20.3% (95% CI: 19.2, 21.5) to 34.1% (95% CI: 32.9, 35.3), the national average remained about the same (17.8% (95% CI: 17.2, 18.4) to 17.3% (95% CI: 16.3, 18.3)). Major differences were also observed in self-reported cost- barriers amongst insured and uninsured middle-income Canadi- ans (Figure 4). While the proportion of insured persons reporting such barriers grew from 6.0% (95% CI: 5.2, 6.9) in 1996 to 17.1% (95% CI: 16.1, 18.1) by 2009, the proportion of uninsured persons who made such reports rose from 14.0%, (95% CI: 13.2, 14.9) to 50.0% (95% CI: 48.7, 51.3). As represented by the indexes of change (data not shown), the percentage of cost-barrier reports grew the fastest among older adults (index of change = 734.4), middle-aged adults (index of change = 366.1), and the uninsured (index of change = 362.3), whereas the respective rise among young adults (index of change = 140.5) and those with higher educational attainment was the lowest (index of change = 172.8). Figure 1. Dental insurance coverage in Canada, by income adequacy and the national average, select years. The proportion of middle-income Canadians with dental insurance increased from 43.1% in 1996 to 48.7% by 2009. Assess to Dental Care for Middle-Income Canadians By 2009, middle-income earners had the lowest levels of dental insurance coverage when compared to all other income groups. While the national average rose steadily between 1996 and 2009, from 55.1% to 68.1%, the proportion of insured middle- income earners declined from 43.1% in 1996 to 40.4% by 2003, then increased to 48.7% by 2009. y doi:10.1371/journal.pone.0057377.g001 having dental insurance compared to their part-time counterparts (72.6% (95% CI: 71.4, 73.8) vs. 64.7% (95% CI: 63.5, 66.0)). having dental insurance compared to their part-time counterparts (72.6% (95% CI: 71.4, 73.8) vs. 64.7% (95% CI: 63.5, 66.0)). Self-reported cost-barriers to dental care Figure 5 shows that the average household expenditures for dental care among middle-income households grew from approx- imately $394 in 1978 to $538 by 2008 (2002 constant dollars). From the early 1990s to the early 2000s, dental expenditures per middle-income household began to rise faster relative to other income levels with the index of change increasing from 94.0 in 1990 to 124.3 by 2000 (data not shown). By 2003, lower-income households reported more expenditures per household compared to middle-income households. Over the same period, higher- income household expenditures remained relatively stable. By 2008, middle-income households had the largest rise in out-of- National data on perceived cost-barriers to dental care were only available for the years 1996, 2003 and 2009. Figure 3 illustrates the percentage of Canadians who reported financial barriers to dental care in select years as a national average and stratified by income adequacy. The percentage of middle-income Canadians who reported cost-barriers increased from 12.6% (95% CI: 11.7, 13.7) in 1996 to 34.1% (95% CI: 32.9, 35.3) by 2009. When compared to all other income groups, middle-income Canadians had the largest rise in the levels of such reports with an index of change of 270.6 by 2009 (data not shown). When comparing the percentage of middle-income Canadians reporting Figure 2. Dental insurance coverage, middle-income and the national average, by work status, select years. When compared to national averages, middle-income earners had lower levels of dental insurance coverage for both full- and part-time workers. Middle-income full- and part-time workers also had the same level of dental insurance in 2009 (47.6% vs. 47.5%), yet nationally, for all incomes, a difference was seen between the two, with a greater percentage of full-time workers having dental insurance compared to their part-time counter- parts (72.6% vs. 64.7%). doi:10.1371/journal.pone.0057377.g002 Figure 3. Self-reported cost-barriers to dental care, by income adequacy the national average, select years. The percentage of middle-income Canadians who reported cost-barriers increased from 12.6% in 1996 to 34.1% by 2009. When comparing the percentage of middle-income Canadians reporting cost-barriers to the national average, a general increase can be seen in both groups from 1996 to 2009. However, the level of growth is much higher among the middle- income group. doi:10.1371/journal.pone.0057377.g003 Figure 3. Self-reported cost-barriers to dental care, by income adequacy the national average, select years. The percentage of middle-income Canadians who reported cost-barriers increased from 12.6% in 1996 to 34.1% by 2009. Data variables and analysis Self-reported dental insurance coverage, cost-barriers to dental care and out-of-pocket expenditures were used as proxies for access to care. All were checked for consistency within and between each survey. Wherever possible, inconsistencies in categorization were minimized by recoding the original variable into a reference set of variables adopted from the CHMS. The February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org 2 Assess to Dental Care for Middle-Income Canadians Trends in dental insurance coverage Again, due to pocket expenditures since 1978 (index of change = 136.7, data not shown). Trends in dental insurance coverage Dental insurance coverage for those in the lowest and lower- middle income levels consistently rose from 1996 to 2009, to the point where they surpassed those of middle-income Canadians. This is likely related to the fact that since 2004, five provincial governments have introduced targeted investments in publicly financed dental care, which has resulted in more dental coverage for eligible low-income Canadians. Again, this has occurred in a context where more than half of middle-income Canadians remain uninsured, with no observed improvements in dental insurance coverage since 1996. Alongside increases to the costs and demand for dental care, and the lack of growth in Canadian incomes, it is no surprise then that middle-income Canadians report increasing difficulties in accessing dental care. In fact, recent national estimates derived from the CHMS demonstrated that middle- income Canadians are over one-and-a-half times more likely to have an unmet dental treatment need than those of the highest incomes [20], and when compared to both high- and low-income families, had the highest mean caries severity scores among their children [15]. Figure 4. Self-reported cost-barriers to dental care among middle-income Canadians, by insurance status, select years. While the proportion of insured persons reporting cost-barriers to dental care grew from 6.0% in 1996 to 17.1% by 2009, the proportion of uninsured persons who made such reports rose from 14.0% to 50.0%. doi:10.1371/journal.pone.0057377.g004 Similar to the findings of Quin˜onez and Grootendorst (2011), this study suggests that changes in the labour market have challenged Canada’s middle-income sector more than the rest [1]. For example, while at the national level the proportion of insured full- and part-time workers has grown, among middle-income workers, whether full- or part-time, levels of insurance have stagnated. Also, historical literature has consistently demonstrated that a greater proportion of full-time workers nationally have dental coverage when compared to part-time workers, yet for middle-income Canadians, no difference was found between these categories. This suggests that middle-income full-time workers have experienced the most drastic changes to their workplace arrangements, which, as discussed in the introduction, stems from decreases to both the quality and availability of employment-based dental insurance. These findings can be relevant to other countries as well, particularly the United States, considering that access to dental care is very similar in both countries given the significant similarities in dental care systems, both of which rely heavily on the presence of employment-based dental insurance. Assess to Dental Care for Middle-Income Canadians Assess to Dental Care for Middle-Income Canadians Figure 4. Self-reported cost-barriers to dental care among middle-income Canadians, by insurance status, select years. While the proportion of insured persons reporting cost-barriers to dental care grew from 6.0% in 1996 to 17.1% by 2009, the proportion of uninsured persons who made such reports rose from 14.0% to 50.0%. doi:10.1371/journal.pone.0057377.g004 Self-reported cost-barriers to dental care When comparing the percentage of middle-income Canadians reporting cost-barriers to the national average, a general increase can be seen in both groups from 1996 to 2009. However, the level of growth is much higher among the middle- income group. doi:10.1371/journal.pone.0057377.g003 Figure 3. Self-reported cost-barriers to dental care, by income adequacy the national average, select years. The percentage of middle-income Canadians who reported cost-barriers increased from 12.6% in 1996 to 34.1% by 2009. When comparing the percentage of middle-income Canadians reporting cost-barriers to the national average, a general increase can be seen in both groups from 1996 to 2009. However, the level of growth is much higher among the middle- income group. doi:10.1371/journal.pone.0057377.g003 Figure 2. Dental insurance coverage, middle-income and the national average, by work status, select years. When compared to national averages, middle-income earners had lower levels of dental insurance coverage for both full- and part-time workers. Middle-income full- and part-time workers also had the same level of dental insurance in 2009 (47.6% vs. 47.5%), yet nationally, for all incomes, a difference was seen between the two, with a greater percentage of full-time workers having dental insurance compared to their part-time counter- parts (72.6% vs. 64.7%). doi:10.1371/journal.pone.0057377.g002 Figure 2. Dental insurance coverage, middle-income and the national average, by work status, select years. When compared to national averages, middle-income earners had lower levels of dental insurance coverage for both full- and part-time workers. Middle-income full- and part-time workers also had the same level of dental insurance in 2009 (47.6% vs. 47.5%), yet nationally, for all incomes, a difference was seen between the two, with a greater percentage of full-time workers having dental insurance compared to their part-time counter- parts (72.6% vs. 64.7%). d i 10 1371/j l 0057377 002 Figure 3. Self-reported cost-barriers to dental care, by income adequacy the national average, select years. The percentage of middle-income Canadians who reported cost-barriers increased from 12.6% in 1996 to 34.1% by 2009. When comparing the percentage of middle-income Canadians reporting cost-barriers to the national average, a general increase can be seen in both groups from 1996 to 2009. However, the level of growth is much higher among the middle- income group. doi:10.1371/journal.pone.0057377.g003 February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org 3 Assess to Dental Care for Middle-Income Canadians Conclusions This study suggests that affordability issues in accessing dental care are no longer just a problem for the lowest income groups in Canada, but now involve middle-income earners as a consequence of their lack of, or decreased access to, comprehensive dental insurance. Assess to Dental Care for Middle-Income Canadians showed that the average expenditures for middle- and low-income households were relatively volatile, while the respective changes for high-income households were far steadier. economic changes, the labour market in the United States has also shifted towards more non-standard jobs, which has resulted in the decline of employment-based insurance for full-time workers from 77% in 1984 to 57% by 1995 [21]. Reports of unmet needs for dental care among middle-income Americans have also increased from 3.9% in 1999 to 5.4% by 2002 [14]. Trends in cost-barriers to dental care Similar to recent work by Thompson (2012), this study found that those from the lower and lower-middle income groups reported the greatest cost-barriers to dental care [22]. However, from 1996 to 2009, the proportion of middle-income Canadians who reported such barriers surpassed that of the lower-middle income group, increasing by approximately three times, resulting in the highest increase among all income levels. This again suggests that changes to dental care plans seen within this time period have had an impact. This is further supported by Thompson (2012), who noted that, while middle-income earners were generally able to access dental professionals, they reported the greatest levels of declining recommended dental treatment due to cost [22]. This study also found that by 2009, three times as many uninsured middle-income Canadians reported cost-barriers to dental care compared to their insured counterparts. This again confirms the fundamental role of dental insurance in accessing dental care in terms of how it mitigates the potential barrier of upfront costs [2,7,22]. Thompson (2012) found that after controlling for other factors, including income, uninsured Cana- dians were almost six times more likely to report cost-barriers to dental care when compared to their insured counterparts [22]. Trends in expenditures for dental care In the absence of adequate dental coverage, individuals are required to spend more money out-of-pocket on dental care (if they ultimately decide to seek care) [22,24]. In this way, out-of- pocket expenditures on dental care can act as a reasonable proxy of access, especially in the context of an insurance rich market such as Canada’s, meaning the more an individual has to spend out-of- pocket, the more difficult it may actually be to obtain care [1,25]. This study found a general increase in average out-of-pocket expenditures on dental care from 1978 to 2008 among all income levels. As Quin˜onez and Grootendorst (2011) have suggested, this is arguably a result of a decline in the robustness of current dental plans, overall increases in dental prices well beyond inflation, and the shift toward demanding more expensive services [1]. Again, the most drastic increases in out-of-pocket expenditures for dental care were found among middle-income Canadians. This study also Limitations This study relied heavily on the availability of consistent data. Although most of the outcomes and socio-demographic variables used were collected reliably over the years as well as between surveys, there were some instances of inconsistency. For example, the years 2005 and 2007 of the CCHS did not provide income adequacy data and the categorical income variable available did not allow for recoding into the desirable format. Dental insurance data was also only collected for some provinces. Therefore, to be more generalizable, these years were excluded. Also, since the current study identified trends among the target population, the results do not provide information on cause-and-effect relation- ships or statistically significant differences. Finally, although this study focused on affordability as proxy of access, it is acknowl- edged that the concept of access encompasses many other factors, including geographic and cultural considerations. Policy implications It can be argued that middle-income Canadians are now facing affordability issues in accessing care comparable to those historically reported by only the lowest income earners. This is of concern, especially since middle-income Canadians have never been on the policy agenda for public assistance to this point. That said, from a public health perspective, some clarity is needed on exactly what is cost-prohibitive, orthodontic and cosmetic dental care, or basic fillings and extractions. Each has its own implications. Importantly, it appears that middle-income seniors also experienced the greatest rise in cost-barriers to dental care. This is not surprising considering that older adults are retaining higher numbers of teeth, thereby potentially increasing their dental needs at a time when they may also be experiencing the diminished income and loss of dental insurance coverage associated with retirement [23]. To be sure, the CHMS found the highest levels of dental disease and the highest rate of no insurance among adults 60–79 years of age [15]. It has also been found that middle-income seniors are the most affected in retirement by the loss of their employment-based dental insurance, as unlike upper-income seniors, they lack sufficient disposable income to afford the cost of dental care [23]. It is also clear that changes in dental care financing are important for the ability of households to access dental care. Hence, as a policy instrument, removing the price barrier to care is likely of most importance [1,26]. As suggested by Thompson (2012), the eligibility criteria for public dental insurance coverage could be altered to include segments of the middle-income population, and/or policies could be made to increase non-wage offers from employers, or to promote the enrolment of workers in employment-based dental insurance plans overall [22]. These policies may, for example, include increasing tax benefits for employers, or mandating the presence of health care benefits in all employment-employee contracts [22]. Whichever way, getting more middle-income earners insured will help to alleviate their cost-barriers to dental care through a reduction in costs at point of purchase. Discussion This study is the first to explore the issue of access to dental care among middle-income Canadians. When compared to all other income levels and to the national average, this study found that, by 2009, middle-income Canadians had the lowest levels of dental insurance coverage and the greatest increase in perceived cost- barriers to dental care. By 2008, middle-income Canadians also had the greatest rise in out-of-pocket dental expenditures since 1978. As proxies of access, this study suggests that access to dental care for this segment of the Canadian population may have become more difficult over the years of observation. Figure 5. Average household expenditures for dental care in Canada, by income adequacy, 1978–2008. The average household expenditures for dental care among middle-income households grew from approximately $394 in 1978 to $538 by 2008 (2002 constant dollars). From the early 1990s to the early 2000s, dental expenditures per middle-income household began to rise faster relative to other income levels. By 2003, lower-income households reported more expenditures per household compared to middle-income households. Over the same period, higher- income household expenditures remained relatively stable. By 2008, middle-income households had the largest rise in out-of-pocket expenditures since 1978. doi:10.1371/journal.pone.0057377.g005 Figure 5. Average household expenditures for dental care in Canada, by income adequacy, 1978–2008. The average household expenditures for dental care among middle-income households grew from approximately $394 in 1978 to $538 by 2008 (2002 constant dollars). From the early 1990s to the early 2000s, dental expenditures per middle-income household began to rise faster relative to other income levels. By 2003, lower-income households reported more expenditures per household compared to middle-income households. Over the same period, higher- income household expenditures remained relatively stable. By 2008, middle-income households had the largest rise in out-of-pocket expenditures since 1978. doi:10.1371/journal.pone.0057377.g005 doi:10.1371/journal.pone.0057377.g005 February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org 4 Assess to Dental Care for Middle-Income Canadians Assess to Dental Care for Middle-Income Canadians References 1. Quin˜onez C, Grootendorst P (2011) Equity in dental care among Canadian households. Int J Equity Health 10:14. Available: http://www.fptdwg.ca/assets/PDF/CHMS/CHMS-E-tech.pdf. Ac- cessed 2012 Oct 25. J q y 2. Locker D, Maggirias J, Quin˜onez C (2011) Income, dental insurance coverage and financial barriers to dental care among Canadian adults. Journal of Public Health Dentistry 71(4): 327–34. 16. Chen WH, Myles J, Picot G (2012) Why have poorer neighbourhoods stagnated economically while the richer have flourished? Neighbourhood income inequality in Canadian cities. Urban Studies 49(4) 877-896. 3. Quin˜onez C, Figueiredo R (2010) Sorry doctor, I can’t afford the root canal, I have a job: Canadian dental care policy and the working poor. Can J Public Health 101(5):481–85. 17. Statistics Canada (2008) Canadian Community Health Survey- Annual Component (CCHS). Available: http://www.statcan.gc.ca/cgi-87bin/imdb/ p2SV.pl?Function=getSurvey&SurvId=3226&SurvVer=1&InstaId=15282& InstaVer=4&SDDS=3226&lang=en&db=imdb&adm=8&dis=2. Accessed 2012 Oct 29. ( ) 4. Muirhead V, Quin˜onez C, Figueiredo R, Locker D (2009) Oral health disparities and food insecurity in working poor Canadians. Community Dentistry and Oral Epidemiology 37: 294–304. 18. Statistics Canada. (2011). Analytical concepts. Available: http://www.statcan.gc. ca/pub/75f0011x/2011001/notes/analytical-analytiques-eng.htm. Accessed: 2012 Oct 29. 5. Muirhead V, Quin˜onez C, Figueiredo R, Locker D (2009) Predictors of dental care utilization among working poor Canadians. Community Dentistry and Oral Epidemiology 37(3): 189–98. 19. Organization for Economic Co-operation and Development (OECD). (2008). OECD indicators. Education at a glance 2008. Available: http://scholar.google. com/scholar?hl=en&btnG=Search&q=intitle:Education+at+a+Glance+2008; +OECD+Indicators#3. Accessed 2012 Oct 30. p gy ( ) 6. Millar WJ, Locker D (1999) Dental insurance and use of dental services. Health Reports 11(1): 55–67. p ( ) 7. Bhatti T, Rana Z, Grootendorst P (2007) Dental insurance, income and the use of dental care in Canada. Journal of Canadian Dental Association 73(1):57. 20. Ramraj C, Azarpazhooh A, Dempster L, Ravaghi V, Quin˜onez C (2012) Dental treatment needs in the Canadian population: analysis of a nationwide cross- sectional survey. BMC Oral Health 12:46. dental care in Canada. Journal of Canadian Dental Association 73(1) 8. Bedos C, Brodeur JM, Benigeri M, Olivier M (2004) Ine´galite´s sociales dans [Social inequalities in the demand for dental care]. Rev Epidemiol Sante Publique 52(3): 261–70 [abstract only]. 21. Wall T, Brown L (2003) Recent trends in dental visits and private dental insurance, 1989 and 1999. Journal of the American Dental Association 134(5):621–7. q ( ) y 9. Arison P (1996) Current trends in dental benefit plans: perspectives on costs of employer-sponsored plans. Ont Dent 73:35–7. 22. Author Contributions Drafted the article and revised it critically for important intellectual content: LS CR CQ. Conceived and designed the experiments: CQ LS. Analyzed the data: LS CQ HL LD. Wrote the paper: CR CQ. February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org 5 Assess to Dental Care for Middle-Income Canadians Assess to Dental Care for Middle-Income Canadians 26. Leake J, Birch S (2008) Public policy and the market for dental services. Comm Dent Oral Epidemiol 36:287–95. References Thompson B (2012) Cost barriers to dental care in Canada. Toronto, ON Graduate Department of the Faculty of Dentistry, University of Toronto. 10. Porter J, Coyte P, Barnsley J, Croxford R (1999) The eff 10. Porter J, Coyte P, Barnsley J, Croxford R (1999) The effects of fee bundling on dental utilization. Health Services Research 34:901–21. 22 p (2 2) , Graduate Department of the Faculty of Dentistry, University of Toronto. 11. Reesor M, Lipsett B (1998) Employer-sponsored health and dental plans- Who is insured? Applied Research Branch, Strategic Policy, Human Resources Development Canada, working paper W–98–2E. 23. Manski RJ, Goodman HS, Reid BC, Macek MD (2004) Dental Insurance Visits and Expenditures Among Older Adults. American Journal of Public Health, 94(5):759–64. 24. Quin˜onez CR (2009) The political economy of dentistry in Canada. Degree of Doctor of Philosophy Dissertation. Toronto, ON: Graduate Department of the Faculty of Dentistry, University of Toronto. 12. Marshall K (2003) Benefits of the job. Perspectives on Labour and Income (Statistics Canada, Catalogue no. 75-001-XIE) (4):5. Available: http://www. statcan.gc.ca/pub/75-001-x/00503/6515-eng.html. Accessed 2012 Oct 25. g p g 13. Zeytinoglu I, Cooke G (2005) Non-standard work and benefits: has anything changed since the Wallace Report? Relations Industrielles 60(1), 29–63. 25. Quin˜onez C, Grootendorst P, Sherret L, Azarpazhooh A, Locker D (2006) An environmental scan of Provincial and Territorial public dental programs in Canada. Toronto, ON: Community Dental Health Services Research Unit, University of Toronto. 14. Blumberg LJ, Holahan J (2004) Changes in insured coverage and access to care for middle-class Americans, 1999–2002. Washington, D.C: Urban Institute. umberg LJ, Holahan J (2004) Changes in insured coverage and access g J J ( ) g g r middle-class Americans, 1999–2002. Washington, D.C: Urban Instit g 15. Health Canada (2010) Report on the Findings of the Oral Health Component of the Canadian Health Measures Survey 2007–2009. Ottawa: Health Canada. 26. Leake J, Birch S (2008) Public policy and the market for dental services. Comm Dent Oral Epidemiol 36:287–95. February 2013 \| Volume 8 \| Issue 2 \| e57377 PLOS ONE \| www.plosone.org PLOS ONE \| www.plosone.org 6
https://openalex.org/W4381300799	https://pubs.rsc.org/en/content/articlepdf/2023/qm/d3qm90041j	English	null	Back cover	Materials chemistry frontiers	2,023	cc-by	381	NEW JOURNAL Uniting disciplines to solve environmental challenges APCs waived until mid-2024 Environmental Science: Advances GOLD OPEN ACCESS NEW JOURNAL Uniting disciplines to solve environmental challenges APCs waived until mid-2024 Environmental Science: Advances GOLD OPEN ACCESS @EnvSciRSC NEW JOURNA rsc li/esadvances Uniting disciplines to solve environmental challenges APCs waived until mid-2024 Environmental Science: Advances GOLD OPEN ACCESS Registered charity number: 207890 @EnvSciRSC Fundamental questions Elemental answers rsc.li/esadvances Uniting disciplines to solve environmental challenges APCs waived until mid-2024 Environmental Science: Advances GOLD OPEN ACCESS Registered charity number: 207890 @EnvSciRSC Fundamental questions Elemental answers rsc.li/esadvances Uniting disciplines to solve environmental challenges APCs waived until mid-2024 Environmental Science: Advances GOLD OPEN ACCESS As featured in: Showcasing research from Prof. S. Nagarajan’s Laboratory, Department of Chemistry, Central University of Tamil Nadu, Thiruvarur, INDIA. Volume 7 \| Number 11 \| 7 June 2023 Registered charity number: 20789 As featured in: Showcasing research from Prof. S. Nagarajan’s Laboratory, Department of Chemistry, Central University of Tamil Nadu, Thiruvarur, INDIA. Acetylene bridged alkoxyphenanthrene and triarylamine-based triads for low threshold voltage with high mobility OFETs New highly conjugated semiconducting triarylamine-based alkoxyphenanthrenes with acetylene bridges were synthesized for high-performance p -channel OFETs. The devices exhibited a carrier mobility of 3.5 cm 2 /Vs with an ON/OFF ratio of 10 8 and the lowest threshold voltage of –3 V. See Samuthira Nagarajan et al ., Mater . Chem . Front ., 2023, 7 , 2225. rsc.li/frontiers-materials MATERIALS CHEMISTRY F R O N T I E R S Volume 7 \| Number 11 \| 7 June 2023 Showcasing research from Prof. S. Nagarajan’s Laboratory, Department of Chemistry, Central University of Tamil Nadu, Thiruvarur, INDIA. Registered charity number: 207890 As featured in: See Samuthira Nagarajan et al ., Mater . Chem . Front ., 2023, 7 , 2225. rsc.li/frontiers-materials MATERIALS CHEMISTRY F R O N T I E R S Volume 7 \| Number 11 \| 7 June 2023 As featured in: Acetylene bridged alkoxyphenanthrene and triarylamine-based triads for low threshold voltage with high mobility OFETs Registered charity number: 207890 New highly conjugated semiconducting triarylamine-based alkoxyphenanthrenes with acetylene bridges were synthesized for high-performance p -channel OFETs. The devices exhibited a carrier mobility of 3.5 cm 2 /Vs with an ON/OFF ratio of 10 8 and the lowest threshold voltage of –3 V. rsc.li/frontiers-materials rsc.li/frontiers-materials
https://openalex.org/W2256732612	https://www.jnma.com.np/jnma/index.php/jnma/article/download/702/1433	English	null	VAGINAL DELIVERY OF THE TRIPLETS	Journal of Nepal Medical Association	2,003	cc-by	1,717	Journal of Nepal Medical Association 2002; 41: 496-498 Journal of Nepal Medical Association 2002; 41: 496-498 CASE REPORT ABSTRACT The spontaneous conception & delivery of triplets occurs in 1: 6889 births. Most of the literature recommends caesarean section for triplets for better perinatal outcome of especially second and third baby, however, we describe vaginally delivered triplets with good perinatal outcome. INTRODUCTION should be the goal for clinicians caring for these women. Greek mythology and the bible give early examples of multiple birth. Romulus and Remus were twins raised by the mother Wolf. Easu and Jacob were the twin boys of Rebekah and Isac. Triplet rate is 1:6889 births. Triplets birth are at much greater risk than singleton in terms of poor birth outcomes. More than 9 of 10 triplet birth are born preterm as compared with < 1 of 10 singleton birta. The average weight of a triplet newborn is about 1.5 kg. The infant death rate for triplets is 12 times1,2 higher that for singleton. Based on their frequency of preterm birth, low birth weight, infant death rate, it is appropriate to characterize all triplet pregnancies high risk.3 Comprehensive and aggressive prenatal care to assure the best outcomes VAGINAL DELIVERY OF THE TRIPLETS Saha R 1, Bastola N 1 JNMA, October - December, 2002, 41 1. Kathmandu Medical College Teaching Hospital, Sinamangal, Kathmandu, Nepal. Address for correspondence : Dr. Rachana Saha, Lecturer Department of Obstetrics and Gynaecology Kathmandu Medical College Teaching Hospital, Sinamangal P. O: 5919, Kathmandu, Nepal CASE REPORT Post delivery blood pressure of mother was 140/100. Babies were transferred to neo-natal intensive care. On admission she was found to be hypertensive with blood pressure of 140/100 mmHg. On obstetric examination the abdomen was hugely distended with multiple parts. The lie of the 1st baby was cephalic, the presentation of the two other babies were difficult to make out. All three fetal heart sound was audible with Doppler. Per vaginal examination showed cervical os dilated up to 5cm, cervical effacement 60%, membrane present and vertex at +1 station. USG showed all the three babies (in three different sac) to be in cephalic labeled as one. Immediately thereafter lie and Foetal Heart Rate of other foetures were checked. Per vaginal examination was done which showed cervical os to be fully dilated, cervical effacement 100% membrane '+' and vertex at +2 station. Rupture of membrane was done which showed presence of clear liquor and second alive male baby weighing 1.485 kg was delivered with apgar score of 6/10, 8/10. The cord was clamped and cut and the baby was labeled as two. The lie and foetal heart rate was rechecked for the third foetus and per vaginal examination was done which showed fully dilated cervical os with cervical effacement (100%), membrane '+' and vertex at +2 station. Artificial rupture of membranes was done which showed clear liquor. Third baby alive female was born after two minutes of the delivery of second baby 1.35 kg with apgar score 6/10,8/10. The cord was clamped and cut and labeled as three. After 15 minutes, the placenta was delivered and inj. Syntocinon 5 IU bolus and 10 IU in drip was kept. Total blood loss was 140 ml. Post delivery blood pressure of presentation and longitudinal lie. The decision was to allow them for vaginal delivery. Blood sample was taken for cross-matching and vein was kept open with Ringer Lactate. Patient was given Cap. Nifedipin 10 mg orally. Fetal heart rate was monitored every half hourly with Doppler. After 4 hours she was examined internally to monitor the progress of labor. The cervix was dilated up to 8cm, cervical effacement was 100%, and vertex was at + 2 station. Artificial rupture of membrane was done which showed clear liquor. At crowning, episiotomy was given. A live male baby weighing 1.43 kg with apgar score of 6/10, 8/10 was born. CASE REPORT 24 years old second gravida at 33 weeks +6days with triplet pregnancy was admitted in active labor on 6 th September, 2002 at Kathmandu Medical College Teaching Hospital. She was diagnosed to have triplet pregnancy from 12 weeks of gestation, by an ultrasonography which showed trichorionic and triamniotic placenta. She had her ante-natal checkup in private clinic. There was no family history of multiple pregnancy or history of ovulation induction. Earlier she had pregnancy loss at 6 months of gestation. JNMA, October - December, 2002, 41 Saha & Bastola Vaginal Delivery of the Triplets 497 admission she was found to be hypertensive h blood pressure of 140/100 mmHg. On obstetric amination the abdomen was hugely distended h multiple parts. The lie of the 1st baby was phalic, the presentation of the two other babies re difficult to make out. All three fetal heart und was audible with Doppler. Per vaginal amination showed cervical os dilated up to 5cm, vical effacement 60%, membrane present and rtex at +1 station. USG showed all the three bies (in three different sac) to be in cephalic sentation and longitudinal lie. The decision was allow them for vaginal delivery. Blood sample s taken for cross-matching and vein was kept en with Ringer Lactate. Patient was given Cap. fedipin 10 mg orally. Fetal heart rate was labeled as one. Immediately thereafter lie and Foetal Heart Rate of other foetures were checked. Per vaginal examination was done which showed cervical os to be fully dilated, cervical effacement 100% membrane '+' and vertex at +2 station. Rupture of membrane was done which showed presence of clear liquor and second alive male baby weighing 1.485 kg was delivered with apgar score of 6/10, 8/10. The cord was clamped and cut and the baby was labeled as two. The lie and foetal heart rate was rechecked for the third foetus and per vaginal examination was done which showed fully dilated cervical os with cervical effacement (100%), membrane '+' and vertex at +2 station. Artificial rupture of membranes was done which showed clear liquor. Third baby alive female was born after two minutes of the delivery of second baby 1.35 kg with apgar score 6/10,8/10. The cord was clamped and cut and labeled as three. After 15 minutes, the placenta was delivered and inj. Syntocinon 5 IU bolus and 10 IU in drip was kept. Total blood loss was 140 ml. CASE REPORT The cord was clamped and cut immediately and The maturity of all babies was 32 weeks. All the babies had good sucking but later they developed jaundice for which they received phototherapy and antibiotics. They were discharged after 27 day with weight of the baby first being 1.75 kg, second baby 1.75 kg, and third baby 1.95 kg. Post partum stay of the mother was uneventful and she was discharged on Cap. Amodipine 5 mg daily. JNMA, October - December, 2002, 41 Saha & Bastola Vaginal Delivery of the Triplets 498 outcomes were similar in both groups.3 Similar observation was done by Ron eir 8 in their management of 19 triplets during the period of 1970-1978. DISCUSSION outcomes were similar in both groups.3 Similar observation was done by Ron eir 8 in their management of 19 triplets during the period of 1970-1978. Labour and delivery of triplets entails increased risk as well. Fetal heart rate monitoring during labor is challenging with vaginal delivery, the first infant is usually born spontaneously or with little manipulation. However, subsequent infants are delivered according to the presenting part, which often require complicated obstetrical maneovres such as total breech extraction with or without podalic version and many even necessitate caesarean delivery . Associated with appositions is increased incidence of cord prolapses and fetal collision. For all these reasons, many clinicians believe that delivery of triplets is best accomplished by caesarean section.3 In this case we allowed vaginal delivery because the triplets were triamniotic, trichorionic, all three babies were in cephalic presentation and they were at 33 week of gestation and in active labour. Vaginal delivery fortunately did not bring any untoward complication in mother and immediate complication to the baby. BIBLIOGRAPHY 1. Keith LG, Oleszcute JJ. Triplets birth in the United States. An epidemic og high risk pregnancy. J. Reprod Med 2002 April 47(4): 259-265. Caesarean section was the mode of delivery in 84% of cases of triplets in the series of 86 triplets delivered in Sweden in 1985 and 1989.4 Similarly more liberal approach towards abdominal delivery of pregnancies of higher fetal number is advocated in the retrospective review of triplets delivered at the Boston perinatal center from 1976 to 1986 because of the increased morbidity from 1st to 2nd and 3rd twin delivered vaginally5 2. Martin JA, Mac Dorman MF, Mathew TJ. Triplets birth trends and outcomes 1971-74. Vital health stat 21 st 1997 Jan: (55); 1:20. 3. Multifoetal pregnancy in Cunningam FG, FG Norman, Heveno KJ: Gilstrap LC, Hanth CJ, Wenstrom DK edited Williams obstetrics (21 st edition), USA, MC-GRAW HILL 2001. 4. Radsted A. Birth of triplets quadruplets in Sweden Nord Med 1991; 106 (11) 297-298. 4. Radsted A. Birth of triplets quadruplets in Sweden Nord Med 1991; 106 (11) 297-298. Lipitz et al6 in their study of 78 triplets from 1975 to 1978 and Growther CA and , Hamilton RA7 in their 105 cases of triplets from 1975-1984 also have recommended caesarean section. 5. Feingold M, Cetrulo C, Peters M, et al. mode of delivery in multiple birth in high order. Acta Genet Med Gemellol (Roma) 1988; 37(1); 105-9. 6. Lipitz S, Reichman B, Paret G, et al. The improving outcome of triplet pregnancies. AM. J Obstet Gynecol 1989 Nov., 161(5): 1979-84. Vaginal delivery is reserved for those circumstances in which survival is not expected because the fetus are markedly immature, or maternal complication make caesarean delivery hazardous to the mother. But then there are other clinicians who believe that vaginal delivery is safe under certain circumstances. For example - Alaniia and colleagues 1998 evaluated a protocol for vaginal delivery. A total of 23 sets of triplets were analyzed and a third were delivered vaginally. Neonatal 7. Growther CA, Hamilton RA. Triplet pregnancy: a ten-year review of 105 cases at Harare maturity hospital Zimbabwe. Acta Genet Med Gemellol (Roma) 1989; 38(3-4): 271-8. 8. Ron-EIR, Caspi E, Schreyer P et al. Triplet and quadruplet preganacies and management. Obstet Gynecol 1981 April 57(4):458-63. ????? JNMA, October - December, 2002, 41
https://openalex.org/W4389939835	https://ejournal.unhasy.ac.id/index.php/nukleosains/article/download/5226/2549	Indonesian	null	VALIDITY OF DEVELOPMENT OF THE GAME WHO WANTS TO BE A MILLIONAIRE COLLOID IN MAN 3 BANJARMASIN	Nukleo Sains	2,023	cc-by	5,525	VALIDITAS PENGEMBANGAN GAME WHO WANTS TO BE A MILLIONAIRE KOLOID DI MAN 3 BANJARMASIN Maslianor1, Ardian Trio Wicaksono2 1,2Program Studi Tadris Kimia, UIN Antasari Banjarmasin, Indonesia Email: maslianor8500@gmail.com DOI: https://doi.org/10.33752/ns.v2i2.5226 Received: 30/11/2023; Revised: 17/12/2023; Accepted: 19/12/2023 Abstract: This study aims to determine the validity of the development of educational game who wants to be a millionaire on colloidal material for class XI students of MAN 3 Banjarmasin with the data source used consists of 1 material expert who is a chemistry teacher and 1 media expert. Data sources used consisted of 1 material expert who is a chemistry teacher and 1 media expert who is a lecturer. who is a lecturer. The type of research used is Research and Development (R&D) model (Define, Design, Develop, and Disseminate). The research results showed that the material expert validation value was 74% and the media expert validation value was 79% so that the game developed could be categorized as valid. Keywords: Validity, Educational game who wants to be a millionaire, Colloid Abstrak: Penelitian ini bertujuan untuk mengetahui validitas dari pengembangan game edukasi who wants to be a millionaire pada materi koloid siswa kelas XI MAN 3 Banjarmasin dengan sumber data yang digunakan terdiri dari 1 ahli materi yang merupakan guru kimia dan 1 ahli media yang merupakan seorang dosen. Jenis penelitian yang digunakan adalah Research and Development (R&D) dengan menggunakan model 4D (Define, Design, Develop, and Disseminate). Hasil penelitian diperoleh bahwa nilai validasi ahli materi sebesar 74% dan validasi ahli media sebesar 79% sehingga game yang dikembangkan dapat dikategorikan valid. Kata kunci: Validitas, Game edukasi who wants to be a millionaire, Koloid PENDAHULUAN pesat dan memberikan dampak besar pada bidang pembelajaran (Agustina & Salsabila 2021). Inovasi teknologi yang Pada zaman sekarang, teknologi telah mengalami perkembangan yang 77 \| e-ISSN 2963-5144 77 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono beragam telah memicu perubahan dalam pendekatan kita terhadap proses belajar- mengajar. Sebagai contoh, penggunaan game edukasi sebagai alat pembelajaran mencerminkan bagaimana teknologi telah diaplikasikan dalam konteks pendidikan. mikroskopik, sehingga menjadikan materi ini sebagai salah satu bagian pembelajaran kimia yang dianggap sulit, dengan kecenderungan tingkat kelulusan siswa yang rendah (Pradilasari dkk 2020). Selama ini, materi koloid masih disampaikan melalui metode ceramah dan penggunaan papan tulis, namun tidak semua konsep dapat dijelaskan dengan baik, menyebabkan kesulitan bagi siswa dalam memahami materi tersebut. Oleh karena itu, seorang guru perlu memiliki keterampilan untuk memotivasi siswa, salah satunya dengan memanfaatkan berbagai media pembelajaran. Pemanfaatan teknologi telah mempermudah siswa dalam menggali pemahaman terhadap materi yang kompleks, seperti pelajaran kimia. Pembelajaran kimia merupakan ilmu, tentang sifat, struktur, komposisi, perubahan, serta energi yang terlibat. Selain itu, dalam pembelajaran kimia, terdapat teori-teori, konsep-konsep, dan hukum-hukum yang digunakan untuk menjelaskan berbagai fenomena alam yang terjadi. (Redhana 2019). Sebagian besar siswa SMA/MA masih menunjukkan kurangnya minat terhadap mata pelajaran kimia, didorong oleh persepsi bahwa pembelajaran kimia dianggap sulit. Pandangan ini berdampak pada motivasi siswa untuk tidak mendalami pembelajaran kimia lebih lanjut. (Budiariawan 2019). Media pembelajaran merupakan sarana yang dimanfaatkan oleh guru untuk menyampaikan informasi kepada siswa secara lebih efektif, memudahkan pemahaman siswa terhadap materi pembelajaran. Penggunaan media tidak hanya berperan sebagai alat bantu, tetapi juga sebagai strategi untuk menjaga perhatian siswa, mencegah kejenuhan, dan menghindarkan rasa bosan selama proses belajar. Pemanfaatan media dalam pembelajaran tidak hanya meningkatkan efisiensi dan efektivitas, tetapi juga mampu memotivasi siswa agar lebih bersemangat dalam mengikuti pembelajaran (Wulandari dkk 2023). Materi koloid merupakan pembelajaran yang tidak melibatkan matematika seperti materi kimia lainnya (Siahaan dkk 2020). Materi koloid mencakup konsep-konsep imajiner dan e-ISSN 2963-5144 \| 78 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … (Tukiran & Rahaju 2017). Game ini dapat dimainkan baik di perangkat Android maupun laptop. Keunggulan utama dari game ini adalah kemampuannya untuk menghindarkan pembelajaran yang monoton, ketersediaan akses yang mudah, fungsinya sebagai alat evaluasi pembelajaran, daya tarik yang menyenangkan, serta kesan berkesan. Game ini dapat dijadikan pilihan untuk dimainkan baik di lingkungan sekolah maupun di rumah. PENDAHULUAN Salah satu sarana yang dimanfaatkan dalam dunia pendidikan adalah game edukasi, yang merupakan jenis permainan yang dirancang khusus untuk memberikan pembelajaran dan pendidikan kepada para pemainnya. Inti dari game edukasi adalah menyajikan materi pembelajaran secara menarik, interaktif, dan menghibur, dengan harapan dapat memudahkan pemahaman serta memberikan motivasi kepada pemain untuk menggali lebih banyak pengetahuan. Dengan pendekatan yang bersifat edukatif namun tetap menghibur, game edukasi bertujuan untuk menggabungkan unsur pendidikan dan hiburan (Yunanto dkk 2019). Pemanfaatan game edukasi sebagai sarana pembelajaran memberikan alternatif menarik bagi guru dalam memperkenalkan, mengajar, dan menguji pemahaman siswa mengenai materi koloid dengan pendekatan yang menyenangkan dan efektif. Salah satu contoh game edukasi yang dapat digunakan untuk membangkitkan minat siswa dalam pembelajaran adalah who wants to be a millionaire. Permainan who wants to be a Berdasarkan observasi awal proses pembelajaran di kelas XI MAN 3 Banjarmasin, terlihat bahwa banyak siswa belum memanfaatkan teknologi secara optimal. Mereka cenderung menggunakan perangkat android dan laptop hanya untuk aktivitas media sosial, permainan yang kurang bermanfaat, dan kegiatan lainnya. Dalam percakapan dengan siswa, terungkap bahwa sebagian besar pembelajaran masih mengandalkan metode ceramah. Guru menggunakan media seperti buku pelajaran, papan tulis, dan presentasi power point, yang menyebabkan siswa merasa jenuh, mengantuk, dan kekurangan minat. Oleh karena itu, diperlukan pendekatan pembelajaran yang lebih menarik dan berbantuan Permainan who wants to be a millionaire merupakan kuis yang terdiri dari berbagai tingkat kesulitan, mulai dari yang mudah hingga paling sulit e-ISSN 2963-5144 79 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono 2015 yang berjudul pengembangan game edukatif tax administration millionaire quiz berbasis adobe flash sebagai media pembelajaran administrasi pajak di kelas XI Akuntansi SMK YPE Sawunggalih Kutoarjo dengan sedikit modifikasi. Proses analisis dilaksanakan dengan menganalisis data hasil validasi materi serta validasi media berdasarkan penilaian angka yang diberikan oleh validator. Skala penilaian yang dipakai dalam penelitian ini dapat ditemukan dalam Tabel 1 berikut. media agar proses pembelajaran menjadi lebih efektif. METODE Jenis penelitian ini adalah penelitian pengembangan atau Research and Development (R&D). Pendekatan penelitian yang akan digunakan adalah pendekatan kualitatif dan pendekatan kuantitatif. Pendekatan ini bertujuan untuk menguji serta meningkatkan produk yang sedang dikembangkan (Yuliani & Banjarnahor 2021). Model yang digunakan dalam penelitian ini adalah Four D models (model 4D). Model 4D adalah singkatan dari Define (definisi), Design (perencanaan), Development (pengembangan), dan Dissemination (penyebaran). Tabel 1. Kategori Penilaian Kategori Skor Sangat Setuju (SS) 5 Setuju (S) 4 Netral (N) 3 Tidak Setuju (TS) 2 Sangat Tidak Setuju (STS) 1 (Sumber: Sanaky dkk 2021) Tabel 1. Kategori Penilaian Kategori Skor Sangat Setuju (SS) 5 Setuju (S) 4 Netral (N) 3 Tidak Setuju (TS) 2 Sangat Tidak Setuju (STS) 1 (Sumber: Sanaky dkk 2021) Pada penelitian ini sumber data berasal dari validator yang terdiri atas satu ahli materi yaitu Ibu Hj. Noormaliana, S.Pd., M.Pd sebagai guru kimia di MAN 3 Banjarmasin, dan satu ahli media yaitu Bapak Alvian Ikhsanul Fatya, S.Pd., M.Si sebagai dosen Program Studi Tadris Kimia di UIN Antasari Banjarmasin. Teknik pengumpulan data pada penelitian ini adalah angket yang mengacu dari skripsi Zulfri Adhi Wibowo mahasiswa Universitas Negeri Yogyakarta tahun Penelitian ini menggunakan instrumen angket lembar penilaian validitas dengan pengumpulan data menggunakan uji validasi yang akan dinilai oleh validator dengan kategori validitas dapat dilihat pada Tabel 2 berikut. Tabel 2 Kategori Tingkat Validitas Persentase (%) Kategori Validitas 81% -100% Sangat Valid 61% - 80% Valid 41% – 60% Cukup Valid 21% – 40% Tidak Valid 0% – 20% Sangat Tidak Valid (Sumber: Wicaksono dkk 2020) e-ISSN 2963-5144 Tabel 2 Kategori Tingkat Validitas Persentase (%) Kategori Validitas 81% -100% Sangat Valid 61% - 80% Valid 41% – 60% Cukup Valid 21% – 40% Tidak Valid 0% – 20% Sangat Tidak Valid (Sumber: Wicaksono dkk 2020) ISSN 2963 5144 Tabel 2 Kategori Tingkat Validitas Persentase (%) Kategori Validitas 81% -100% Sangat Valid 61% - 80% Valid 41% – 60% Cukup Valid 21% – 40% Tidak Valid 0% – 20% Sangat Tidak Valid (Sumber: Wicaksono dkk 2020) Tabel 2 Kategori Tingkat Validitas \| 80 e-ISSN 2963-5144 \| 80 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … Penentuan nilai validitas sebagai berikut: Hasil Validitas = 𝑡𝑜𝑡𝑎𝑙 𝑠𝑘𝑜𝑟 𝑦𝑎𝑛𝑔 𝑑𝑖𝑝𝑒𝑟𝑜𝑙𝑒ℎ 𝑠𝑘𝑜𝑟 𝑚𝑎𝑘𝑠𝑖𝑚𝑎𝑙 x 100% ini membantu untuk memahami akar permasalahan yang ada dalam konteks pembelajaran kimia. HASIL DAN PEMBAHASAN Validasi bertujuan untuk mengevaluasi produk melalui analisis oleh para ahli, guna mendapatkan rekomendasi serta masukan yang berguna dalam mengukur mutu produk. Sebelum dilakukan validasi, terlebih dahulu dilakukan proses pengembangan produk. Proses pengembangan produk menggunakan model Research and Development (R&D), dengan hasil akhir berupa game edukasi who wants to be a millionaire yang berfokus pada materi koloid. Model pengembangan yang digunakan dalam penelitian ini adalah model 4D, yang merupakan singkatan dari Define, Design, Development, dan Dissemination (Harjanto dkk 2022). Langkah-langkah dalam proses pengembangan produk menggunakan model pengembangan 4D adalah sebagai berikut: Berdasarkan hasil observasi dan wawancara, terlihat bahwa pembelajaran di kelas masih umum dilakukan melalui metode ceramah dan diskusi, hal ini membuat siswa lebih banyak mendengarkan dan memahami penjelasan dari guru. Di MAN 3 Banjarmasin khususnya untuk kelas XI IPA 3 belum ada bahan ajar tambahan, seperti penggunaan game edukasi who wants to be a millionaire pada materi koloid. Game edukasi dapat menjadi pilihan yang efektif bagi pendidik dalam meningkatkan minat belajar siswa (Karseno dkk 2021). Apabila game ini dikembangkan dengan baik, diharapkan bisa menjadi media pembelajaran yang sangat menarik, efektif, dan juga menjadikan teknologi dapat dimanfaatkan dengan baik. Penentuan nilai validitas sebagai berikut: Hasil Validitas = 𝑡𝑜𝑡𝑎𝑙 𝑠𝑘𝑜𝑟 𝑦𝑎𝑛𝑔 𝑑𝑖𝑝𝑒𝑟𝑜𝑙𝑒ℎ 𝑠𝑘𝑜𝑟 𝑚𝑎𝑘𝑠𝑖𝑚𝑎𝑙 x 100% Hasil Validitas = 𝑡𝑜𝑡𝑎𝑙 𝑠𝑘𝑜𝑟 𝑦𝑎𝑛𝑔 𝑑𝑖𝑝𝑒𝑟𝑜𝑙𝑒ℎ 𝑠𝑘𝑜𝑟 𝑚𝑎𝑘𝑠𝑖𝑚𝑎𝑙 x 100% METODE Pada tahap ini memiliki 4 cara menganalisis yaitu analisis awal akhir, spesifikasi tujuan pembelajaran, spesifikasi tujuan pembelajaran, dan spesifikasi tujuan pembelajaran (Triana 2020). Pembahasaan dari tahap-tahap analisis sebagai berikut: Tahap Define Tahap ini tujuan untuk secara rinci mendefinisikan kebutuhan yang esensial dalam proses pembelajaran, sekaligus mengumpulkan data dan informasi terkait dengan pengembangan produk yang akan dilakukan (Ana 2018). Proses Berdasarkan hasil dari wawancara dan observasi, terlihat bahwa siswa 81 \| e-ISSN 2963-5144 81 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono untuk SMA/MA XI semester 2” ditulis oleh Aisyah yang diterbitkan pada tahun 2018, buku ajar “kimia koloid dan permukaan” ditulis oleh Arneli dan Yuyuk Astuti yang diterbitkan pada tahun 2019, dan juga pada buku ajar “kimia untuk SMA/MA XI ( kelompok peminat matematika dan ilmu alam)” ditulis oleh Unggul Sudarmo yang diterbitkan pada tahun 2014. untuk SMA/MA XI semester 2” ditulis oleh Aisyah yang diterbitkan pada tahun 2018, buku ajar “kimia koloid dan permukaan” ditulis oleh Arneli dan Yuyuk Astuti yang diterbitkan pada tahun 2019, dan juga pada buku ajar “kimia untuk SMA/MA XI ( kelompok peminat matematika dan ilmu alam)” ditulis oleh Unggul Sudarmo yang diterbitkan pada tahun 2014. menunjukkan kurangnya minat dalam mengikuti proses pembelajaran kimia. Hal ini kemungkinan terkait dengan persepsi umum bahwa mata pelajaran tersebut dianggap rumit. Selain itu, media pembelajaran yang digunakan saat ini tidak berhasil menarik minat siswa dan cenderung membosankan. Jika sumber pembelajaran dapat diubah sedemikian rupa sehingga lebih menarik dan menyenangkan bagi mereka, dapat diharapkan semangat siswa dalam memahami pelajaran kimia akan meningkat. Salah satu contoh media pembelajaran yang dapat membangkitkan minat siswa adalah penggunaan game edukasi yang interaktif (Pratama dkk 2019). Maka, dalam peran sebagai peneliti yang menangani permasalahan ini, peneliti menciptakan sebuah media pembelajaran yang berupa game edukasi who wants to be a millionaire dengan fokus pada materi koloid. Tujuan Pembelajaran ditetapkan berdasarkan turunan indikator yang diperoleh dari Kompetensi Inti (KI) dan Kompetensi Dasar (KD) yang tercantum dalam kurikulum 2013 dan juga ada tambahan tujuan pembelajaran dari peneliti untuk menyesuaikan dengan isi materi. Tujuan pembelajaran yang digunakan pada game edukasi who wants to be a millionaire adalah: (1) Siswa diharapkan dapat menjelaskan pengertian koloid; (2) Siswa diharapkan dapat menjelaskan perbedaan koloid, suspensi dan larutan; (3) Siswa diharapkan dapat mengelompokkan jenis koloid; (4) Siswa diharapkan dapat menjelaskan sifat-sifat koloid; (5) Siswa diharapkan dapat membedakan koloid liofil dan koloid liofob; (6) Siswa diharapkan dapat menjelaskan cara pembuatan koloid; (7) Siswa diharapkan Dalam analisis konsep, materi koloid dibagi menjadi tiga sub bab, yaitu penjelasan tentang sistem koloid dan jenisnya, sifat-sifat koloid, serta proses pembuatan dan pemurnian koloid. Tahap Define Isi materi yang terdapat pada produk game edukasi who wants to be a millionaire mengacu pada modul pengayaan “kimia e-ISSN 2963-5144 \| 82 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … koloid, serta daftar pustaka yang digunakan dalam materi dan kuis. dapat menjelaskan cara pemurnian koloid. Penyusunan pengembangan game edukasi who wants to be a millionaire mengenai materi koloid melibatkan langkah-langkah berikut. Pertama, desain game disusun melalui aplikasi powerpoint. Kemudian, rancangan tersebut diubah menjadi aplikasi android menggunakan ispring suite dan website 2 APK builder pro. Ispring suite memudahkan pembuatan media pembelajaran interaktif tanpa memerlukan keahlian dalam bahasa pemrograman. Aplikasi ini juga memungkinkan konversi desain ke berbagai format yang diinginkan (Larasati dkk 2022). Vol.2, No.2, Juli - Desember 2023 Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono menggunakan powerpoint diubah menggunakan powerpoint show. Melakukan revisi tahap pertama pada game edukasi who wants to be a millionaire pada materi koloid untuk dilakukan perbaikan produk yang dibuat berdasarkan saran dan komentar oleh dosen pembimbing. Adapun saran dan komentar dosen pembimbing tentang produk yang dikembangkan adalah: Soal game edukasi who wants to be a millionaire sebaiknya dikurangi yang awalnya 60 soal menjadi 15 soal agar waktu untuk uji coba game sesuai dengan rencana; dan dalam menu game edukasi who wants to be a millionaire , sebaiknya tombol profil dan petunjuk diletakkan di bagian paling atas agar siswa dapat melihatnya lebih awal sebelum memulai membaca materi dan kuis. Letak tombol profil dan petunjuk di bagian menu game dapat dilihat pada gambar 1 berikut. Tahap Design Tahap ini bertujuan untuk menciptakan suatu kerangka atau konsep perangkat pembelajaran yang akan digunakan dalam proses pembelajaran (Kurniawan & Dewi 2017). Peneliti memulai proses pengembangan rancangan produk dengan langkah pertama yaitu mengidentifikasi Kompetensi Inti (KI), Kompetensi Dasar (KD), dan Indikator yang akan menjadi dasar. Setelah menganalisis elemen- elemen tersebut, peneliti kemudian melanjutkan membuat desain produk, kemudian merancang produk yang mencakup berbagai komponen seperti intro game, menu, petunjuk penggunaan, profil peneliti, dan pada setiap materi terdapat Kompetensi Inti (KI), Kompetensi Dasar (KD), serta tujuan pembelajaran. Produk yang direncanakan juga memiliki tiga materi utama terkait koloid, yakni materi 1 yang membahas pengertian dan jenis-jenis koloid, materi 2 yang membahas sifat- sifat koloid, dan materi 3 yang membahas pembuatan dan pemurnian koloid. Selain itu, produk juga mencakup kuis game berdasarkan materi koloid, video materi yang berkaitan dengan Tahap ini bertujuan untuk menciptakan suatu kerangka atau konsep perangkat pembelajaran yang akan digunakan dalam proses pembelajaran (Kurniawan & Dewi 2017). Peneliti memulai proses pengembangan rancangan produk dengan langkah pertama yaitu mengidentifikasi Kompetensi Inti (KI), Kompetensi Dasar (KD), dan Indikator yang akan menjadi dasar. Setelah menganalisis elemen- elemen tersebut, peneliti kemudian melanjutkan membuat desain produk, kemudian merancang produk yang mencakup berbagai komponen seperti intro game, menu, petunjuk penggunaan, profil peneliti, dan pada setiap materi terdapat Kompetensi Inti (KI), Kompetensi Dasar (KD), serta tujuan pembelajaran. Produk yang direncanakan juga memiliki tiga materi utama terkait koloid, yakni materi 1 yang membahas pengertian dan jenis-jenis koloid, materi 2 yang membahas sifat- sifat koloid, dan materi 3 yang membahas pembuatan dan pemurnian koloid. Selain itu, produk juga mencakup kuis game berdasarkan materi koloid, video materi yang berkaitan dengan Website 2 APK builder pro merupakan sebuah perangkat lunak yang memiliki kemampuan untuk mengonversi file powerpoint dan ispring suite menjadi aplikasi android. Dengan menggunakan aplikasi ini, proses konversi dilakukan secara efisien, memungkinkan transformasi desain pembelajaran interaktif menjadi aplikasi yang dapat diakses melalui platform android (Hadi 2020). Di samping itu, untuk versi aplikasi pada laptop, hasil rancangan pengembangan game edukasi who wants to be a millionaire e-ISSN 2963-5144 e-ISSN 2963-5144 83 \| Vol.2, No.2, Juli - Desember 2023 Tahap Development Proses perkembangan terdiri dari kelima tahapan berikut: pertama, konsultasi dengan dosen pembimbing; kedua, melakukan revisi tahap pertama; ketiga, melakukan validasi ahli materi dan ahli media; keempat, melaksanakan revisi tahap kedua; dan terakhir, respon siswa. Langkah pertama dalam pengembangan penelitian melibatkan perancangan game edukasi who wants to be a millionaire setelah melakukan diskusi mendalam dengan dosen pembimbing mengenai materi, perumusan soal kuis, dan desain secara menyeluruh. Hasil dari kerjasama ini berhasil menciptakan produk awal game edukasi who wants to be a millionaire yang difokuskan pada pemahaman materi koloid. Gambar 1. Letak Tombol Profil dan Petunjuk di Bagian Menu Game Gambar 1. Letak Tombol Profil dan Petunjuk di Bagian Menu Game \| 84 e-ISSN 2963-5144 Validity Of Development Of … Validity Of Development Of … Vol.2, No.2, Juli - Desember 2023 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … Validasi merupakan tahap penilaian yang digunakan untuk menilai sejauh mana desain produk dapat dianggap sebagai suatu pendekatan yang rasional dan efektif dalam konteks media pembelajaran. Validasi pada tahap ini dilakukan berdasarkan pertimbangan rasional, bukan berdasarkan bukti empiris dari pengalaman lapangan (Putri dkk 2021). Tujuan dari proses validasi adalah untuk mengevaluasi produk melalui analisis oleh para ahli, dengan harapan mendapatkan rekomendasi dan masukan yang bermanfaat dalam mengukur kualitas produk yang telah dikembangkan. Dalam melakukan penilaian melalui kuesioner dengan skala Likert, penilaian dilakukan berdasarkan kriteria berikut: 1 = sangat tidak setuju, 2 = tidak setuju, 3 = netral, 4 = setuju, dan 5 = sangat setuju. Validasi materi dilakukan oleh Ibu Hj. Noormaliana, S.Pd., M.Pd yaitu guru kimia di MAN 3 Banjarmasin sebagai ahli materi. Validasi oleh ahli materi bertujuan untuk mengukur sejauh mana produk yang sedang dikembangkan memiliki tingkat kelayakan yang memadai, khususnya dalam hal aspek materi (Fallensky dkk 2021). Penilaian produk oleh ahli materi ditinjau dan dinilai dari tiga aspek, yaitu aspek materi, aspek kebahasaan, dan aspek soal dengan 21 indikator. Hasil penilaian penilaian dari validasi ahli materi dapat dilihat pada Tabel 3 berikut. Tabel 3 Keseluruhan Hasil Penilaian dari Validasi Ahli Materi Tabel 3 Keseluruhan Hasil Penilaian dari Validasi Ahli Materi Aspek Jumlah Rata-rata Persentase Kategori Materi 41 3,72 74,5% Valid Kebahasaan 12 4 80% Valid Soal 25 3,57 71% Valid Jumlah Keseluruhan 78 Rata-Rata Keseluruhan 3,71 Persentase Keseluruhan 74% Kategori Valid Pada Tabel 3 yang membahas Konversi Skor Data Kuantitatif, dapat dilihat bahwa pada aspek materi memiliki persentase 74,5%, aspek kebahasaan memiliki persentase 80%, aspek soal memiliki persentase 71%, dan untuk rata-rata persentase keseluruhan adalah 74%. Ini menunjukkan bahwa produk yang dikembangkan masuk ke dalam kategori “Valid”. Penilaian dari ahli materi bahwa produk yang dikembangkan masih kurang sempurna. Hal ini disebabkan karena: Pada materi dan soal hanya berupa tulisan; dan Soal lebih bervariasi disesuaikan dengan tingkat-tingkat kesulitan (soal e-ISSN 2963-5144 e-ISSN 2963-5144 85 \| Maslianor, Wicaksono Vol.2, No.2, Juli - Desember 2023 minat belajar mereka (Saputro 2019). Sedangkan berdasarkan penelitian Defi dan Faiza menyatakan bahwa Penggunaan video animasi menjadi suatu alat yang sangat efektif dalam konteks pembelajaran karena dapat menarik perhatian siswa, meningkatkan kemampuan mereka untuk mengingat informasi, serta memfasilitasi visualisasi yang lebih baik terkait dengan konsep, objek, dan hubungan-hubungannya dengan cara yang jelas dan menarik (Defi & Faiza 2021). Validity Of Development Of … mencangkup soal mudah, sedang, dan sulit). Solusi yang dilakukan peneliti agar produk yang dikembangkan menjadi lebih baik adalah: Pada materi dan soal, penggunaan gambar dan video (seperti video animasi) sebaiknya dimasukkan untuk meningkatkan daya tarik dan pemahaman materi; dan Soal berurutan dari pengertian sampai pemurnian koloid. Berdasarkan pada poin bahwa penggunaan gambar dan video pada materi dan soal di dalam produk dapat meningkatkan daya tarik dan pemahaman materi. Hal tersebut sesuai dengan penelitian Safitri dan Kabiba yang menyatakan bahwa memanfaatkan media gambar yang memadai menjadi kunci utama untuk mengoptimalkan proses pembelajaran. Hal ini tercermin dalam peningkatan minat belajar siswa, yang pada akhirnya memberikan kontribusi positif terhadap hasil belajar mereka (Safitri & Kabiba 2020). Hal tersebut diperkuat dengan diperkuat dengan hasil penelitian Saputro menyatakan bahwa Penggunaan media gambar dalam pembelajaran tidak hanya efektif dalam meningkatkan partisipasi siswa, tetapi juga dapat merangsang Validasi media dilakukan oleh yaitu Bapak Alvian Ikhsanul Fatya, S.Pd., M.Si yaitu dosen prodi Tadris kimia di UIN Antasari Banjarmasin sebagai ahli media. Validasi oleh ahli media bertujuan untuk mengukur sejauh mana produk yang sedang dikembangkan memiliki tingkat kelayakan yang memadai, khususnya dalam hal aspek media Fallensky dkk 2021). Penilaian produk oleh ahli media ditinjau dan dinilai dari dua aspek, yaitu mencakup aspek rekayasa perangkat lunak dan aspek komunikasi visual dengan 22 indikator. Hasil penilaian penilaian dari validasi ahli media dapat dilihat pada Tabel 4 berikut. e-ISSN 2963-5144 \| 86 Vol.2, No.2, Juli - Desember 2023 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … Validity Of Development Of … Tabel 4. Keseluruhan Hasil Penilaian dari Validasi Ahli Media Aspek Jumlah Rata-rata Persentase Kategori Rekayasa Perangkat Lunak 41 3,72 74,5% Valid Komunikasi Visual 46 4,18 84% Sangat Valid Jumlah Keseluruhan 87 Rata-Rata Keseluruhan 3,95 Persentase Keseluruhan 79% Kategori Valid Tabel 4. Keseluruhan Hasil Penilaian dari Validasi Ahli Media Solusi yang dilakukan peneliti agar produk yang dikembangkan menjadi lebih baik adalah: (1) Perbaikan sistem pada produk; (2) Bantuan menjawab pada kuis dihilangkan; (3) Pada materi dan soal, penggunaan gambar dan video (seperti video animasi) sebaiknya dimasukkan untuk meningkatkan daya tarik dan pemahaman materi. Pada Tabel 4 yang membahas konversi skor data kuantitatif, dapat dilihat bahwa pada aspek rekayasa perangkat lunak memiliki persentase 74,5% dinyatakan “Valid”, aspek komunikasi visual memiliki persentase 84% dinyatakan “Sangat Valid”, dan untuk rata-rata persentase keseluruhan adalah 79% yang menunjukkan bahwa produk yang dikembangkan masuk ke dalam kategori “Valid”. Validity Of Development Of … Penilaian dari ahli media bahwa produk yang dikembangkan masih kurang sempurna. Hal ini disebabkan karena: (1) Aplikasi mengalami keterlambatan dalam eksekusi, dan informasi skor untuk setiap pertanyaan tidak dapat ditampilkan. Hal ini dapat disebabkan oleh perubahan dalam sistem yang terjadi saat proses menyalin dan memindahkan file dari flashdisk ke laptop maupun sebaliknya; (2) Selalu dapat 1 miliar saat penggunaan; (3) Pada opsi bantuan bisa dipakai lebih dari 1x dari setiap pertanyaan; (4) Pada materi dan soal hanya berupa tulisan. Berdasarkan pada poin bahwa penggunaan gambar dan video pada materi dan soal di dalam produk dapat meningkatkan daya tarik dan pemahaman materi. Hal tersebut sesuai dengan hasil penelitian Siregar yang dinyatakan bahwa Penggunaan gambar dalam pembelajaran dapat secara nyata meningkatkan pencapaian akademis siswa, karena gambar mampu menangkap perhatian siswa dan memperkuat konsentrasi mereka pada materi yang diajarkan oleh guru (Siregar 2017). Hal tersebut diperkuat dengan diperkuat dengan hasil penelitian Nurhayati menyatakan bahwa Pemanfaatan media gambar dalam 87 \| e-ISSN 2963-5144 87 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono proses belajar-mengajar memberikan insentif kepada siswa untuk menjadi lebih tertarik, memahami, dan ikut berpartisipasi secara aktif dalam pembelajaran (Nurhayati 2022). Sedangkan berdasarkan hasil penelitian Suherman dkk (2022) yang menyatakan bahwa Penggunaan media pembelajaran berbasis video animasi diyakini dapat menghasilkan peningkatan dalam hasil belajar, motivasi siswa, keterampilan berbicara, dan berbagai aspek lainnya yang berkaitan dengan proses pembelajaran (Suherman dkk 2022). Setelah melakukan revisi berdasarkan saran dan komentar dari validator, dilakukan penilaian respons siswa terhadap produk yang dikembangkan. Penilaian respon siswa ini di lakukan di sekolah MAN 3 Banjarmasin, siswa yang menjadi responden yaitu siswa kelas XI IPA 3 yang berjumlah 31 siswa. Langkah ini diambil untuk memahami bagaimana siswa merespons game edukasi who wants to be a millionaire pada materi koloid. Validity Of Development Of … Validity Of Development Of … Wants To Be A Millionaire pada Materi Koloid Siswa Kelas XI MAN 3 Banjarmasin, dapat disimpulkan bahwa hasil validasi dari para ahli, baik dalam aspek materi maupun media, menunjukkan pencapaian yang positif. Validasi oleh ahli materi mencapai rata- rata 74% dengan penilaian "Valid", sementara validasi oleh ahli media mencapai rata-rata 79% dengan kriteria "Valid". Hasil ini menandakan bahwa game edukasi who wants to be a millionaire yang telah dikembangkan memiliki tingkat validitas yang baik dan dapat dianggap sebagai sarana pembelajaran yang layak. diambil sebagai tanggapan terhadap pembatasan waktu yang tidak efisien, yang menghambat kemampuan peneliti untuk mempelajari dan mengunggah aplikasi ke platform Play Store. Langkah ini dipicu oleh beberapa siswa yang mengalami kesulitan dalam melakukan instalasi aplikasi setelah mengunduhnya melalui whatsapp. Tujuan lainnya adalah untuk mempermudah proses pengunduhan aplikasi bagi siswa, mengurangi kendala teknis, serta meningkatkan keterjangkauan aplikasi pembelajaran. KESIMPULAN Berdasarkan hasil Validasi dari Pengembangan Game Edukasi Who https://doi.org/10.36088/islamika.v 3i1.1047 Arkadiantika, I., Ramansyah, W., Effindi, M.A., & Dellia, P. 2020. Pengembangan Media Pembelajaran Virtual Reality pada Materi Pengenalan Termination dan Splicing Fiber Optic. Jurnal Dimensi Pendidikan dan Pembelajaran, Special Issue, SEMNASDIKJAR 2019, (January, 2020): 30-36, https://doi.org/10.24269/dpp.v0i0.2 298. https://doi.org/10.36088/islamika.v 3i1.1047 Arkadiantika, I., Ramansyah, W., Effindi, M.A., & Dellia, P. 2020. Pengembangan Media Pembelajaran Virtual Reality pada Materi Pengenalan Termination dan Splicing Fiber Optic. Jurnal Dimensi Pendidikan dan Pembelajaran, Special Issue, SEMNASDIKJAR 2019, (January, 2020): 30-36, https://doi.org/10.24269/dpp.v0i0.2 298. Tahap Dissemination Melakukan revisi tahap kedua pada game edukasi who wants to be a millionaire pada materi koloid untuk dilakukan penyempurnaan produk yang dibuat berdasarkan saran dan komentar oleh validator. Setelah melalui validasi oleh ahli materi dan ahli media, game edukasi who wants to be a millionaire yang telah dikembangkan terbukti layak untuk diujicoba. Meskipun demikian, perlu dilakukan revisi sesuai dengan komentar dan saran yang diberikan oleh ahli materi dan ahli media sebelum melanjutkan ke tahap pengujian. Selanjutnya peneliti berkonsultasi dan mendapatkan izin dari dosen pembimbing untuk bisa langsung uji coba produk akhir kepada siswa. Setelah melalui tahap revisi dalam pengembangan, langkah berikutnya adalah menyebarluaskan produk yang telah dibuat. Diperlukan upaya untuk menyebarkan dan memperkenalkan produk tersebut kepada siswa yang lebih besar di luar lingkup pengembangannya sendiri (Arkadiantika dkk, 2020). Proses penyebaran produk ini dilakukan di MAN 3 Banjarmasin, khususnya di kelas XI IPA. Penggunaan android dan laptop di sekolah telah diizinkan di sekolah MAN 3 Banjarmasin. Penyebaran aplikasi game edukasi who wants to be a millionaire dilakukan melalui whatsapp dan flashdisk kepada siswa dan guru mata pelajaran kimia. Keputusan ini e-ISSN 2963-5144 e-ISSN 2963-5144 \| 88 Vol.2, No.2, Juli - Desember 2023 DAFTAR RUJUKAN Ana, R. F. R. 2018. Penggunaan Model Four D dalam Pengembangan Bahan Ajar Mata Kuliah Perencanaan Pembelajaran pada Mahasiswa STKIP PGRI Tulungagung. PEDAGOGY, Vol 5 (2) :64-74, https//ejournal.upm.ac.id/index.php /pedagogy/article/view/14 Agustina, N., & Salsabila, U. H. 2021. Peran Teknologi Pendidikan dalam Pembelajaran. ISLAMIKA, Vol 3 (1):123–33, Ana, R. F. R. 2018. Penggunaan Model Four D dalam Pengembangan Bahan Ajar Mata Kuliah Perencanaan Pembelajaran pada Mahasiswa STKIP PGRI Tulungagung. PEDAGOGY, Vol 5 (2) :64-74, https//ejournal.upm.ac.id/index.php /pedagogy/article/view/14 Agustina, N., & Salsabila, U. H. 2021. Peran Teknologi Pendidikan dalam Pembelajaran. ISLAMIKA, Vol 3 (1):123–33, e-ISSN 2963-5144 89 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono Sekolah Dasar. Jurnal Teknologi Pembelajaran Indonesia, Vol 11 (1):16-25, https://doi.org/10.23887/jurnal_tp.v 11i1.62 Kurniawan, D., & Dewi, S.V. 2017. Pengembangan Perangkat Pembelajaran Dengan Media Screencast-O-Matic Mata Kuliah Kalkulus 2 Menggunakan Model 4- D Thiagarajan. Jurnal Siliwangi Seri Pendidikan, Vol 3 (1): 214- 219, https://doi.org/10.37058/jspen didikan.v3i1.193 Larasati, D., Wrahatnolo, T., Rijanto, T., & Anifah, L. 2022. Pengembangan Media Pembelajaran Ispring Suite 9 Berbasis Android Pada Mata Pelajaran Dasar Listrik Dan Elektronika Di SMK Negeri 3 Surabaya. Jurnal Pendidikan Teknik Elektro, Vol 11 (1): 79- 85, https://doi.org/10.26740/jpte.v1 1n01.p79-85 Nurhayati. 2022. Penerapan Media Gambar untuk Meningkatkan Hasil Belajar IPA pada Siswa Kelas VI SDN 011 Sungai Salak. Primary: Jurnal Pendidikan Guru Sekolah Dasar, Vol 11 (3): 908, http://dx.doi.org/10.33578/jpfkip.v 11i3.8965. Budiariawan, I.P. 2019. Hubungan Motivasi Belajar Dengan Hasil Belajar Pada Mata Pelajaran Kimia. Jurnal Pendidikan Kimia Indonesia, Vol 3 (2)): 103, https://doi.org/10.23887/jpk.v3i2.2 Budiariawan, I.P. 2019. Hubungan Motivasi Belajar Dengan Hasil Belajar Pada Mata Pelajaran Kimia. Jurnal Pendidikan Kimia Indonesia, Vol 3 (2)): 103, https://doi.org/10.23887/jpk.v3i2.2 1242 Defi, A. N., & Faiza, D. 2021. Pengembangan Media Pembelajaran Berbasis Video Animasi Pada Mata Pelajaran Dasar Listrik Elektronika. Jurnal Vocational Teknik Elektronik Dan Informatika, Vol 9 (2): 113- 118, https://doi.org/10.24036/votet eknika.v9i2.112046 Harjanto, A. A., Rustandi., & Caroline, J.A. 2022. Implementasi Model Pengembangan 4D dalam Mengembangkan Media Pembelajaran Berbasi Online pada Mata Pelajaran Pemrograman Web di SMK Negeri 7 Samarinda. Jurnal SIMADA (Sistem Informasi dan Manajemen Basis Data), Vol 5 (2): 2-12, https://doi.org/10.30873/simada.v5i Vol 3 (2)): 103, https://doi.org/10.23887/jpk.v3i2.2 1242 Defi, A. N., & Faiza, D. 2021. Pengembangan Media Pembelajaran Berbasis Video Animasi Pada Mata Pelajaran Dasar Listrik Elektronika. Jurnal Vocational Teknik Elektronik Dan Informatika, Vol 9 (2): 113- 118, https://doi.org/10.24036/votet eknika.v9i2.112046 Harjanto, A. A., Rustandi., & Caroline, J.A. 2022. Implementasi Model Pengembangan 4D dalam Mengembangkan Media Pembelajaran Berbasi Online pada Mata Pelajaran Pemrograman Web di SMK Negeri 7 Samarinda. DAFTAR RUJUKAN Jurnal SIMADA (Sistem Informasi dan Manajemen Basis Data), Vol 5 (2): 2-12, https://doi.org/10.30873/simada.v5i 2.341. Karseno., Sariyasa., & Astawan, I.G. 2021. Pengembangan Media Game Edukasi Berbasis Android pada Topik Bilangan Bulat Kelas VI Nurhayati. 2022. Penerapan Media Gambar untuk Meningkatkan Hasil Belajar IPA pada Siswa Kelas VI SDN 011 Sungai Salak. Primary: Jurnal Pendidikan Guru Sekolah Dasar, Vol 11 (3): 908, http://dx.doi.org/10.33578/jpfkip.v 11i3.8965. Nurhayati. 2022. Penerapan Media Gambar untuk Meningkatkan Hasil Belajar IPA pada Siswa Kelas VI SDN 011 Sungai Salak. Primary: Jurnal Pendidikan Guru Sekolah Dasar, Vol 11 (3): 908, http://dx.doi.org/10.33578/jpfkip.v 11i3.8965. Karseno., Sariyasa., & Astawan, I.G. 2021. Pengembangan Media Game Edukasi Berbasis Android pada Topik Bilangan Bulat Kelas VI \| 90 e-ISSN 2963-5144 Vol.2, No.2, Juli - Desember 2023 Validity Of Development Of … Pradilasari, L., Gani, A., & Khaldun, I. 2020. Pengembangan Media Pembelajaran Berbasis Audio Visual pada Materi Koloid Untuk Meningkatkan Motivasi dan Hasil Belajar Siswa SMA. Jurnal Pendidikan Sains Indonesia, Vol 7 (1): 9–15, https://doi.org/10.24815/jpsi.v7i1.1 3293. Pratama, L.D., Bahauddin, A., & Lestari, W. 2019. Game Edukasi: Apakah Membuat Belajar Lebih Menarik?. At-Ta’lim Jurnal Pendidikan, Vol 5 (1): 39-50, https://doi.org/10.36835/attalim.v5i 1.64. Sanaky, M.M., Saleh, M., &Titaley., H.D. 2021. Analisis Faktor-Faktor Penyebab Keterlambatan Pada Proyek Pembangunan Gedung Asrama MAN 1 Tulehu Maluku Tengah. Jurnal Simetrik. Vol 11 (1): 432-439, https://doi.org/10.31959/js.v11i1.61 Putri, N. K., Yuberti., & Hasanah, U. 2021. Pengembangan Media Pembelajaran Berbasis Web Google Sites Materi Hukum Newton pada Gerak Benda. Physics and Science Education Journal (PSEJ), Vol 1 (3): 133-143, https://doi.org/10.30631/psej.v1i3.1 033. Redhana, I. W. 2019. Mengembangkan Keterampilan Abad Ke-21 dalam Pembelajaran Kimia. Jurnal Inovasi Pendidikan Kimia, Vol 13 (1): 2239-2253, Putri, N. K., Yuberti., & Hasanah, U. 2021. Pengembangan Media Pembelajaran Berbasis Web Google Sites Materi Hukum Newton pada Gerak Benda. Physics and Science Education Journal (PSEJ), Vol 1 (3): 133-143, https://doi.org/10.30631/psej.v1i3.1 Putri, N. K., Yuberti., & Hasanah, U. 2021. Pengembangan Media Pembelajaran Berbasis Web Google Sites Materi Hukum Newton pada Gerak Benda. Physics and Science Education Journal (PSEJ), Vol 1 (3): 133-143, https://doi.org/10.30631/psej.v1i3.1 033. Redhana, I. W. 2019. Mengembangkan Keterampilan Abad Ke-21 dalam Pembelajaran Kimia. Jurnal Inovasi Pendidikan Kimia, Vol 13 (1): https://doi.org/10.31959/js.v11i1.61 Saputro, L. E. 2019. Pengaruh Penggunaan Media Gambar terhadap Hasil Belajar Siswa Kelas IV. Pedagogi: Jurnal Ilmu Pendidikan, Vol 19 (1): 37-43, https://doi.org/10.24036?fif.100.v19 i1.326.000-000. Saputro, L. E. 2019. Pengaruh Penggunaan Media Gambar terhadap Hasil Belajar Siswa Kelas IV. Pedagogi: Jurnal Ilmu Pendidikan, Vol 19 (1): 37-43, https://doi.org/10.24036?fif.100.v19 i1.326.000-000. Siahaan., Andri, K.W., Simangunsong, A.D., Nainggolan, L.L., & Simanjuntak. M.A. 2020. Pengembangan Bahan Ajar Kimia Materi Koloid Untuk SMA Dengan e-ISSN 2963-5144 91 \| 91 \| Vol.2, No.2, Juli - Desember 2023 Maslianor, Wicaksono Model Inkuiri Terbimbing Dengan Media Animasi. Jurnal Nalar Pendidikan, Vol 8 (2): 130, https://doi.org/10.26858/jnp.v8i2.15 376. Siregar, R. 2017. Penggunaan Media Gambar untuk Meningkatkan Hasil Belajar IPA pada Siswa Sekolah Dasa. Suara Guru : Jurnal Ilmu Pendidikan Sosial, Sains, dan Humaniora, Vol 3 (4): 27- 31, https://doi.org/10.26714/jps.8.1. 2020.27-31. Suherman., Rahmani, A., & Alpiani. 2020. Pengembangan Media Pembelajaran Berbasis Video Animasi untuk Meningkatkan Minat Belajar Siswa pada Materi Panas dan Perpindahannya di Kelas V Sekolah Dasar. Jurnal Pendidikan Dasar Setia Budhi, Vol 6 (1): 1-7, https://stkipsetiabudhi.e- journal.id/jpd. Tukiran., &Rahaju. 2017. Penerapan Model Pembelajaran Teams Games Tournament Berbantuan Permainan Model Inkuiri Terbimbing Dengan Media Animasi. Jurnal Nalar Pendidikan, Vol 8 (2): 130, https://doi.org/10.26858/jnp.v8i2.15 376. Model Inkuiri Terbimbing Dengan Media Animasi. Jurnal Nalar Pendidikan, Vol 8 (2): 130, https://doi.org/10.26858/jnp.v8i2.15 376. Triana, A. 2020. Pengembangan Bahan Ajar Modul Berbasis Contextual Learning Kelas IV SD/MI. Skripsi, Universitas Islam Negeri Raden Intan Lampung . Triana, A. 2020. Pengembangan Bahan Ajar Modul Berbasis Contextual Learning Kelas IV SD/MI. Skripsi, Universitas Islam Negeri Raden Intan Lampung . Wicaksono, A.A.,, Sunarti, T. & Zainuddin, A. 2020. Validitas dan Reliabilitas Instrumen Penilaian Kinerja Praktikum Pada Materi Alat Optik Kelas XI SMA. IPF : Inovasi Pendidikan Fisika, Vol 9 (3): 325- 331, https://doi.org/10.26740/ipf.v9 3 325 331 Wicaksono, A.A.,, Sunarti, T. & Zainuddin, A. 2020. Validitas dan Reliabilitas Instrumen Penilaian Kinerja Praktikum Pada Materi Alat Optik Kelas XI SMA. IPF : Inovasi Pendidikan Fisika, Vol 9 (3): 325- 331, https://doi.org/10.26740/ipf.v9 n3.p325-331. Suherman., Rahmani, A., & Alpiani. 2020. Pengembangan Media Pembelajaran Berbasis Video Animasi untuk Meningkatkan Minat Belajar Siswa pada Materi Panas dan Perpindahannya di Kelas V Sekolah Dasar. Jurnal Pendidikan Dasar Setia Budhi, Vol 6 (1): 1-7, https://stkipsetiabudhi.e- journal.id/jpd. Wulandari, A. P., Salsabila, A.A., Cahyani, K., Nurazizah, T.S., & Ulfiah, Z. 2023. Pentingnya Media Pembelajaran dalam Proses Belajar Mengajar. Journal on Education, Vol 5 (2): 28-36, https://doi.org/10.31004/joe.v5i2.10 74 Belajar Siswa pada Materi Panas dan Yuliani, W., & Banjarnahor, N. 2021. Metode Penelitian Pengembangan (Rnd) Dalam Bimbingan Dan Konseling. Quanta: IKIP SILIWANGI, STBA YAPARI ABA Bandung, Vol 5 (3): 111-118, https://doi.org/10.22460/q.v2i1p21- 30.642. Tukiran., &Rahaju. 2017. Penerapan Model Pembelajaran Teams Games Tournament Berbantuan Permainan Who Wants To Be a mMillionaire. Math Didactic: Jurnal Pendidikan Matematika, Vol 3 (3): 188-198, https://doi.org/10.33654/math.v3i3. 71. Yunanto, A. A., Herumurti, D., Rochimah, S., & Kuswardayan, I. 2019. English Education Game \| 92 e-ISSN 2963-5144 Validity Of Development Of … Vol.2, No.2, Juli - Desember 2023 Using Non-Player Character Based On Natural Language Processing. Procedia Computer Science 161: 502-508, https://doi.org/10.1016/j.procs.2019 .11.158. Using Non-Player Character Based On Natural Language Processing. Procedia Computer Science 161: 502-508, https://doi.org/10.1016/j.procs.2019 .11.158. Using Non-Player Character Based On Natural Language Processing. Procedia Computer Science 161: 502-508, https://doi.org/10.1016/j.procs.2019 .11.158. https://doi.org/10.1016/j.procs.2019 .11.158. 93 \| e-ISSN 2963-5144 e-ISSN 2963-5144 93 \|

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